BR102016029628A2 - ANTIMICROBIAN COMPOSITION, METHOD FOR PREVENTION, MITIGATION AND CONTROL OF BIOGENIC PROCESS AND BACTERIAL GROWTH, AND USE OF COMPOSITION - Google Patents

Abstract

"Antimicrobial composition, methods for prevention, mitigation and control of biogenic processes and bacterial growth, and use of the composition" The present invention comprises antimicrobial compositions comprising 1% to 98% (weight / volume) of at least one selected bioactive compound between a compound of formula (I) and / or a compound of formula (Iia) and / or a compound of formula (Iib): formula (Iia) formula (Iib). The present invention further relates to methods for prevention, mitigation and control of biogenic processes, for inhibiting bacterial growth and the use of said composition for the manufacture of a medicament for inhibiting bacterial targets, for the prevention, mitigation and control of biogenic processes, preferably souring, microorganism-influenced corrosion and oil well capping, or inhibition of degradation of corrosion inhibitors in diesel transport pipelines, as well as asepsis, disinfection and sterilization of domestic, hospital and industrial environments, including prevention contamination and reduction of infection in a hospital environment.

Description

"ANTIMICROBIAL COMPOSITION, METHODS FOR PREVENTION, MITIGATION AND CONTROL OF BIOGENIC PROCESSES AND GROWTH OF BACTERIA, AND, USE OF THE COMPOSITION"

FIELD OF APPLICATION OF THE INVENTION [0001] The present invention is related to the control of microbial growth in industrial or service segments, more specifically to Antimicrobial Compositions for application in the oil industry and in the environmental and health areas.

FUNDAMENTALS OF THE INVENTION [0002] The offshore oil industry daily processes millions of cubic meters of seawater for injection into oil deposits, with the aim of repressurizing it, stimulating oil recovery and, thus, contributing to increased production. [0003] There are rules, applied to oil companies, that fix the maximum allowed concentration for bacteria present in seawater treatment systems for injection, both in planktonic form, as well as in sessile form, that is, capable of form biofilm. The purpose of this control is to avoid problems caused by bacterial growth, which is favored by the anaerobic environment, rich in sulfate and nutrients, existing inside the secondary oil recovery system.

[0004] The growth of sulfate-reducing anaerobic bacteria (BRS) is well known in seawater treatment systems for injection. This bacterial action is a consequence, among other factors, of the stimulation by the metabolic input associated with the high concentration of ion sulfate present in seawater, its preferred electron acceptor for breathing in anaerobic environments. As the main cause of this sulfate-reducing metabolism, the occurrence of a souring process (acidification) is identified, which is characterized by the intensive production of hydrogen sulfide (H ₂ S) in petroleum reservoirs submitted to an improved oil recovery method, disqualifying the quality standard of the fluids produced, oil and natural gas, and increasing the respective treatment costs with the oily water produced.

2/113 [0005] The highly corrosive characteristics of the H ₂ S result in the replacement of metallic materials for columns, pipes, oil and gas storage tanks and other equipment. Serious environmental and work safety problems occur in oil production facilities due to the high toxicity of H ₂ S, requiring constant monitoring of the atmosphere and the use of individual and collective protection equipment.

[0006] The classic biochemical equation (Postgate, J.R., The sulphate reducing bacterium, 2nd edition, 1984), which expresses BRS metabolism, is as follows:

moles lactate + 1 moles SO4 ' ² + 2 moles acetate + 2 moles CO ₂ + 2 moles H2O + 1 moles S' ² [0007] The growth dynamics of these populations leads to the production of a high gradient of H ₂ S and a drop in redox potential at the interface with the metal, in addition to the production of other aggressive acid metabolites (carbon dioxide and organic acids), pitting inducers (localized corrosion) and precursors of localized corrosion processes that are difficult to predict and control. BRSs, intercropped with other anaerobic and / or facultative organisms, usually in the presence of iron sulfide precipitates (FeS), of varying compositions and stoichiometries, whether or not they are protective of metallic substrates, are usually present.

[0008] This biogenic process is associated with the adhesion of biofilms to the internal carbon steel surfaces of seawater injection systems.

[0009] Biofilms are formed by bacterial consortia, in which BRS is the key organism, causing the phenomenon of corrosion influenced by microorganisms (CIM) (COUTINHO, CMLM “Ultrastructural and cytochemical study of bacterial biofilms involved in the process of microbiological corrosion in Petrobras offshore platforms. ”Master's Dissertation. IOC. Fiocruz, Rio de Janeiro, 1991).

[0010] Anaerobiosis, a requirement of the BRS group, and its relatively restricted nutrition, ensure that these organisms are always found as components of a microbial community or consortium.

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These consortia are often found in the form of biofilms, on interfaces or on solid substrates, and allow the creation of anaerobic microenvironments even in an environment characterized as aerobic.

[0011] When adhesion occurs in the surface area of mineral substrates of the oil rock, in the vicinity of injection wells or producers, the plugging of the pores of the rock may hinder the flow of sea water, preventing the success of the recovery method applied, further enhancing the occurrence of souring phenomena.

[0012] Furthermore, it is widely recognized that H ₂ S presents a high degree of risk to human health with the description of acute (death) and chronic episodes (disabling occupational diseases and also risks to populations exposed to the contaminated urban atmosphere), demonstrating intangibility as to its quantification.

[0013] The scenarios described thus determine the potential for the occurrence of biogenic processes that are difficult to control, which generate problems that compromise the success of oil production, with serious impacts on the environment and job security.

[0014] As a consequence, in order to meet the quality standards and criteria required by the oil industry, integrating them with work safety and protection of the marine environment, it is essential to control microbial growth targeting BRS, in consortium with other anaerobic and / or optional organisms, in significantly high volumes of sea water to be injected into reservoirs located in deep waters.

[0015] The preventive measure traditionally adopted to inhibit the growth of these bacteria, initially present in planktonic form, consists of the application of commercial biocides with internationally reported uses in the oil industry.

[0016] Biocides are chemical agents, present as active ingredients in the compositions, usually with a broad spectrum of activity. The classification of these products covers, among others, disinfectants for human use, public and private health, veterinary hygiene,

4/113 food and drinking water, preservatives, pest control and others, including antifouling products with biological scale inhibition characteristics.

[0017] According to the Biocides Directive (98/8 / EC), "biocidal products are intended to destroy, prevent action or exert a controlling effect on any harmful organism by biological or chemical means". Annex V of that document contains the types of products and also their distinction: antibiotic, when the antimicrobial acts against infections in humans and / or animals and the action of biocides such as sterilants, disinfectants, antiseptics, preservatives .

[0018] Particularly in relation to the oil industry, the most suitable biocides are made up of organic-based molecules, with non-oxidizing characteristics, usually associated with solvents, surfactants, chelating substances and other adjuvants. The dosage of these products is carried out with determined frequency (shock and / or continuous) in concentrations of the order of parts per million-volume (ppm-v) in points of the seawater injection system, usually downstream of the deaerating tower and / or in the tower itself.

[0019] The specific characteristics of these biocides, facing the challenge of the target microorganisms, are known, as, for example, the evidenced, in the case of two relevant examples, in Videla, Η. A. et al “Microbiologically influenced corrosion: looking to the future”, International Microbiology, 2005, 8: 169-180:

1. Tetrakis (hydroxymethyl) phosphonium sulfate (THPS): bactericidal claimed as specific for BRS and of low environmental toxicity for use in seawater injection systems.

2. Glutaraldehyde (GA): recognized use in industrial water treatment, in the concentration range from 10 ppm-v to 70 ppm-v, against algae, fungi, bacteria and biofilms. In addition, glutaraldehyde is claimed to be effective in removing biofilms and controlling the growth of metabolically active organisms in dynamic laboratory systems that simulated the conditions of water injection systems for oil recovery. Field tests showed a reduction in planktonic BRS to levels not

5/113 detectable, two hours after treatment with 50 ppm-v of glutaraldehyde, corresponding to an injection water flow of 3.8 x 10 ³ m ³ / day (Eager, RG et al. “Paper Number 84”, CORROSION 88, 1988, NACE, USA).

[0020] With the growth of environmental demands, even in the 90s, it was pointed out that chemical products applied in offshore units in the North Sea, defined as aggressive to the marine environment and workers' health, according to standardized tests, were included in a confidential document entitled Black List, with the provision of their replacement, within a specified period, for non-toxic products, the “green products” [Bos, A., “Regulatory developments with respect to the Harmonized Mandatory Control System for offshore chemicals and their implications for the development of environmentally friendlier products ”, 1997, Technical Publication - SERVO DELDEN BV], [0021] Currently, it is recognized that commercial biocides, the uses of which have traditionally been reported in the oil industry, are via-deregra, toxic from the point of view of their impacts on human health and the environment. Examples of commercial biocides are listed in TABLE 1, with the respective product links in Haz-Map Database. TABLE 1

COMMERCIAL BIOCIDES

Glutaraldehyde (GA)

THPS tetrakis (hydroxymethyl) sulfate

DAZOMET (3,5-dimethyl-1,3,5-thiadiazinan2-thione)

DBNPA (2,2-dibromo-3-nitrilepropionamide

ISOTIAZOLINONE (S)

I- Methylisothiazolinone

II- Chloromethylisothiazole

III- Benzisothiazolinone___________________

ANTRAQUINONE (9,10-anthracenedione) * Source: Haz-Map database

6/113 [0022] Haz-Map is an important database of occupational health and toxicology maintained by the United States government, containing guidance and information on the adverse effects associated with exposure to chemical and biological agents in places of job. The respective links show the adverse effects on health and the environment of the biocides referred to. Among the various effects caused by the biocides mentioned, skin sensitization or irritation is common to all.

[0023] Glutaraldehyde and DBNPA also cause irritation to the eyes, nose and throat. Workers exposed to glutaraldehyde (in diluted solutions) and isothiazolinones, respectively, acquired occupational asthma. Dazomet causes eye irritation and inhalation of DBNPA, as well as THPS (in high concentration), in addition to sensitization, can lead to pneumonia and pulmonary edema. Anthraquinone is reported as possibly carcinogenic to humans [(Group 2B) - IARC 2012: Anthraquinone], [0024] In the environment, glutaraldehyde is considered to present a lower potential risk insofar as high dilutions can be achieved due to its favorable characteristics of biodegradation (OECD 301 D) and non-bioaccumulation, for example, in aquatic environments. However, such a compound proves to be highly toxic to the standard freshwater invertebrate organism, Daphnia magna, an important link in the aquatic food chain, according to US EPA 660 / 3-75-009 (US Environmental Protection Agency Standard) [0025] increasing environmental restrictions and the need to attend to aspects of work safety, as well as to maintain the effectiveness in inhibiting microorganisms of interest to the industry, particularly oil, there is a growing and current search for efficient and less toxic molecules (green molecules) for biofilm control in the oil industry, considering that the breakdown of biofilm may be the key to control the harmful effects caused by undesirable microorganisms (Albuquerque, AC & Andrade, CA & Neves, B. “Biocorrosion - from the integrity of the biofilm to the integrity of the material ”. Corros. Prot. Mater. (2014) .Vol. 33, Nos 1).

7/113 [0026] In the specialized literature, the references mentioned below were researched and found, which propose the use of combinations of substances with additional or complementary properties to the known biocidal substances themselves, as an alternative to the use of the said commercial biocides. per se:

[0027] Reduction of the dose of traditional biocide, such as glutaraldehyde (GA) or tetrakis (hydroxymethyl) phosphonium (THPS) sulfate, through its association with another auxiliary substance said to be less toxic, p. eg “green additives” (chelating agents, biodegradable) such as:

1. EDDS - ethylenediaminodisuccinate (Wen, Jie; Gu, Tingyue, “Green biocide enhancers enhanced the biocide inhibition of the growth of sulfate reducing bacteria.” AlChE Annual Meeting, Conference Proceedings, San Francisco, CA, United States (2006) Nov. 12-17);

2. HEIDA disodium salt (N- (2-hydroxyethyl) iminodiacetic acid) [J. Wen, K. Zhao, T. Gu ,, I. Raad, “Chelators enhanced biocide inhibition of planktonic sulfate-reducing bacterial growth” World J Microbiol Biotechnol (2010) 26: 1053-1057];

3. EDDS - ethylenediaminodisuccinate and methanol (J. Wen, D. Xu, T. Gu, I. Raad “A green triple biocide cocktail consisting of a biocide, EDDS and methanol for the mitigation of planktonic and sessile sulfate-reducing bacteria” World J Microbiol Biotechnol (2012) 28: 431-435).

4. EDDS (ethylene diaminosuccinate) and methanol (J. Wen ET AL “Biocide cocktail consisting of glutaraldehyde, ethylene diamine disuccinate (EDDS), and methanol for the mitigation of souring and biocorrosion” Corrosion: (2012) November 994-1002.

[0028] These associations claim the potentiation regarding the antimicrobial action against BRS, particularly regarding the inhibition of the growth of its planktonic and sessile forms (reference 1 mentioned in §0029), planktonic (reference 2 mentioned in §0030) and also the removal of biofilms established by that bacterium (reference 3 mentioned in §0031) and inhibition of biofilm establishment and pitting due to Microorganism Influenced Corrosion (MIC) (reference 4 mentioned in §0032).

8/113 [0029] Other alternatives to associations with traditional biocides have also been proposed, comprising:

a) Joint application of the biocide THPS and the substance anthraquinone in treatment units and storage of oily salty ballasts, with prospects for success in the control of H ₂ S and cost-benefit adequacy (Burger, Andrade, Rebello & Ribeiro. Flexible Treatment Program for Controlling H ₂ S in FPSO Produced-Water Tanks, 2007 and Penkala et al, 2004)

b) Equimolecular mixture of amino acids D-tyrosine, D-methionine, Dtryptophan and D-leucine added to a biocide cocktail containing THPS and ethylenediamino-N, N'-disuccinic acid (EDDS) resulted in a triple biocidal cocktail much more effective than THPS and EDDS isolated or mixed. This mixture prevents the formation of biofilm and removes the formed film (D. Xu “D-amino acids for the enhancement of a binary biocide cocktail consisting of THPS and EDDS a bgainst an SRB biofilm” World J Microbiol, Biotechnol (2012) 28: 1641-1646).

c) Synergistic combination of biocide THPS and D-tyrosine (found in many food proteins) prevents the formation of a biofilm on the surface of the C1018 carbon steel coupon in batch treatment tests. Removes biofilms from BRS established on the coupon surface in shock treatment tests (D. Xu ET AL 'A synergistic D-tyrosine and tetrakis hydroxymethyl phosphonium sulfate biocide combination for the mitigation of SRB biofilm. ”World J Microbiol Biotechnol (2012) 28 : 3067-3074).

d) Synergistic compositions - US20090186858 (07/23/2009) comprising THP salt and a biopenetrant, which is a polymer of an unsaturated carboxylic acid or a copolymer of unsaturated carboxylic acid with a sulfonic acid, said polymer and copolymer being finished by group unsaturated mono- or di-phosphonated carboxylic acid or having such monomers incorporated in the structure (spine) of the polymer. These compositions act synergistically by increasing the biocidal efficacy of the THP salt against both forms [planktonic bacteria (free-swimming) and sessile (attached to a substrate)] and also to

9/113 effectiveness of the THP salt in dissolving FeS precipitate. The THP salt includes THPS.

e) Biocides for well stimulation and fluid treatment US2010218950 (02/09/2010) - comprising a treatment and / or well stimulation fluid that includes water, other additives and a biocide consisting of Dazomet (3,5-dimethyl -1,3,5-thiadiazinan-2-thione) in an effective amount to inhibit bacterial growth and minimize antagonistic reactions between the biocide and other additives. Well injection compositions are also described.

f) Biocidal compositions and methods of use WO2011059532 (19/05/2011) - comprising: 2,2-dibromo-3-nitrilopropionamide (DBNPA) and a compound selected from the group consisting of 1 (3-chloroalyl) -3.5 , 7-triaza-1-azoniaadamantano CTAC); tris (hydroxymethyl) nitromethane and a hexahydrotriazine compound. Such compositions are useful for microbial control in aqueous or water-containing systems. One of the objectives of this association is to increase the duration of action of the biocide DBNPA.

g) Biocidal compositions based on glutaraldehyde and methods of use WO2010138420 (02/12/2010) - such compositions being synergistic for the control of microorganisms in aqueous systems comprising glutaraldehyde and a compound selected from a group consisting of 1- (3-chloroallyl) ) -3,5,7-triaza-1-azoniaadamantano (CTAC); tris (hydroxymethyl) -nitromethane; and a hexahydrotriazine. The observed synergistic effect allows the use of smaller quantities to achieve biocidal properties, thus reducing the environmental impact and material cost.

h) The compositions of the documents (f) and (g), respectively, associate with commercial biocides the compound 1- (3-chloroallyl) -3,5,7-triaza-1azoniaadamantane (CTAC), microbicide for use in secondary injection of oil whose toxic characteristics are contained in the document RED Facts - EPA 738-95-016 - April 1995 accessed through Haz-Map. In these two cases, compatibility between the respective components of these biocidal compositions would result in an increase in the respective toxicities.

10/113 [0030] On the other hand, integrating different fields for potential use, it appears that, in the areas of health and environment, biocides play a very important role in the prevention of infectious processes in different scenarios. These substances are applied as surface disinfectants, where phenolic compounds and quaternary ammonium salts stand out, as disinfectants and surgical sterilizers (asepsis of surgical environments), instruments and medical equipment, with the use of enzyme-based detergents , and the substances peracetic acid, orthophthalaldehyde and glutaraldehyde, and alcoholic preparations for general use.

[0031] Among the alternatives proposed to traditional biocides, the transformation of THPS, due to its low corrosion inhibition, into an active surface agent through the coupling reaction with different fatty acids (decanoic, dodecanoic, palmitic and stearic) stands out. . The resulting compounds, in this case, showed an increase in the biocidal effect and in the mechanism of inhibition against corrosion of carbon steel, in acidic medium, because they act as inhibitors of the mixed type that prevent the cathodic and anodic parts of the corrosion reaction in acidic medium.

[0032] Another line of alternatives, aimed at meeting environmental restrictions, comprises the association of known products of natural origin with traditional commercial biocides. This is the case for the biocidal formulations of US2013 / 0029884 (01/31/2013) - Biocide formulations - said to be environmentally friendly and safe for the individuals who handle them and for the materials (equipment) with which they maintain contact. Used in aqueous oil and gas drilling fluids, in the treatment of aqueous sludge in mining operations, they also apply to injection water, and include:

the water

b) at least one biocide selected from the group:

i) a halogen-substituted xylenol compound ii) a phenolic compound iii) an essential or natural antimicrobial oil iv) an antimicrobial component of essential or natural oil

v) combinations of at least two of (i), (ii), (iii) and (iv)

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c) at least one anionic, amphoteric, non-ionic surfactant and mixtures thereof

d) at least one solubilizing agent [0033] This formulation comprises all of its components, because each one is always present. There are several options for the biocide, such as: phenolic compound including monophenols and substituted phenols; lemongrass essential oil and citral, the subject of an article reported below. However, the preferred biocidal substance, which is present in almost all of the examples, is the compound parachloromethaxylenol (PCMX). It should be noted that parachloromethane-xylenol (PCMX) is identified as a cause of allergic contact dermatitis in Haz-Map and severe eye irritation (Category I, indicative of the highest degree of acute toxicity) according to EPA - US Environmental Protection Agency.

[0034] The process to prevent or mitigate bio-encrustation [W02011100189 - 18/08/2011 - Process for preventing ormitigating biofouling] proposes to associate the use of cinnamaldehyde additive with traditional commercial biocides to accentuate the biocidal effect of said compounds aiming at the prevention, or the reduction of the occurrence of biological fouling in industrial water systems, including fluids associated with oil and gas exploration and production (for example, fluids used for secondary and tertiary recovery). Example 1 of this process shows the synergistic action of the association of cinnamaldehyde with the commercial biocide THPS on samples containing BRS without establishing the degree of biocidal efficiency achieved. In example 2 of this process, the action of the additive cinnamaldehyde is tested, in isolation, on culture obtained from a water sample containing unidentified bacteria present in incrustation taken from an oil production site and introduced in synthetic brine. The evaluation of biocidal activity, in this case, was carried out by measuring turbidity (600 nm), using a wide absorbance scale that shows a reduction in turbidity with an increase in cinnamaldehyde concentration. This reduction in the concentration of bacterial cells was attributed to this indirect measure, disregarding the fact that the effectiveness of death is best proven by classical microbiology methods, with the specificity defined against the target to be controlled, which

12/113 presents a determinant for a biocide specially designed for the oil industry.

[0035] They are proposed as environmentally friendly alternatives to the use of commercial synthetic biocides in the petroleum industry, the essential oil of lemon grass, and its main component, citral. Both inhibit the growth of planktonic and sessile BRS. Both can be used in formulations in the same way as synthetic biocides, in water, methanol and isopropanol [Korenblum E. et al “Antimicrobial action and anti-corrosion effect against sulfate reducing bacteria by lemongrass (Cymbopogon citratus) essential oil and its major component , the citral ”AMB Express 2013, 3:44], Citral consists of a mixture of two aldehydes of the same molecular composition (Ο-ιοΗιθΟ), of different structures, geranial and neral, presenting itself as a liquid with a strong lemon odor with toxicity characteristics as it is reported as irritating to the skin and mucous membranes. It is able to be absorbed by this tissue as indicated in the Haz-Map, causing, in higher concentrations (> 8%), allergic dermatitis. Therefore, special care is required to mitigate risks in the handling of this substance.

[0036] In the state of the art, in the various proposals alternative to traditional commercial biocides, there was no evidence of antimicrobial efficiency proven to challenge different BRS target cultures originating from different sites in the oil industry, both surface and subsurface, of ships and other facilities, showing population reduction - control in a short time. There was also no evidence of biocidal action against Total Heterotrophic Anaerobic Bacteria (BANHT) consortium with BRS and also the measurable control of generalized corrosion. On the other hand, the possibility of expanding the spectrum of antimicrobial bioactivity was not considered in defiance of strains of environmental bacteria, some of which are also common to the oil industry. It is observed that, despite the scientific efforts made in the studies listed, the challenge of finding effective antimicrobial substances in their biocidal action and performance, for use in the oil industry, still being able to minimize risks to the environment and the environment. human health (Green Products).

13/113 [0037] In summary, it is considered relevant to consider that, despite the activity reported against target microorganisms, biocides traditionally used in the oil industry have a toxic character to man and the environment and do not always act in inhibiting corrosion.

[0038] The referred “green alternatives” to traditional biocides, in most cases, comprise the reduction of the dose of traditional commercial biocide, glutaraldehyde and THPS, for example, through the association of these with compounds that would potentiate the respective antimicrobial action against BRS , both in planktonic and sessile form. In addition, such compositions also propose to act in the prevention of formation, as well as in the removal of biofilms established by this same bacterium, claiming even to increase the efficiency in the dissolution of precipitates of iron sulfide.

[0039] The aforementioned proposed alternatives come to demonstrate that, even on a smaller scale, the toxic effect remains, with cases even including proposed compositions associating traditional biocidal substances with compounds also endowed with microbicidal and toxic characteristics, signaling that, even obtaining a synergistic effect, the purpose of reducing the environmental impact would be limited.

[0040] It should be noted that there is also a clear trend in the sense that biocide compositions, according to recent proposals, aiming at meeting environmental restrictions and safety requirements, add to the commercial biocides substances of natural origin known for their popular use. In some cases, it is proposed that these natural products also act as biocides as an alternative to commercial biocides, but it is noted that the limitations due to toxicity persist. Thus, there remains a need for a biocidal composition that fully meets the requirements for effective and specific antimicrobial action as to the targets to be controlled, in conjunction with the environmentally friendly characteristics and the health of the worker.

[0041] In addition to the requirements of the industry, particularly oil, there is a need for alternatives aimed at operating in the food and pharmaceutical industries, as well as in outpatient and outpatient settings.

14/113 hospitals, veterinary facilities, and in domestic use, especially as surface disinfectants.

[0042] In this context, we highlight the use of compositions containing synthetic phenolic compounds, usually halogenated, and quaternary ammonium salts. Especially in the health area, as surgical sterilizers for medical instruments and equipment, they are additionally associated with enzyme-based detergents, as well as with traditional biocidal and / or antiseptic substances, such as peracetic acid, orthophthalaldehyde, glutaraldehyde, and alcoholic preparations of general use. The HSE (Health and Safety Executive / UK), the official body of the Health System in England, developed a critical study regarding the disinfection of endoscopes, having come to a strong conclusion that “there is no single system for disinfecting endoscopes and no disinfectant. more apropriate". This source also addresses the concern about serious problems related to occupational asthma due to the use of glutaraldehyde (GA), since this biocidal substance has been commonly applied in hospital environments. It also mentions that GA would be associated with the fifth cause of asthma, according to data from health agencies (The Health and Safety Commission / HSC), and it was recommended to reduce exposure to that substance in workplaces in order to comply with regulations established (Control of Substances Hazardous to Health / COSHH). According to the same sources, commercial products based on GA (trade names: CIDEX, Activated Glutaraldehyde Solution, ASEP, Totacide) were gradually withdrawn from supply stocks in England (UK) and, in mid-2005, ceased. fully use it.

[0043] On the other hand, in the 2014 WHO report, based on data from 114 countries, it highlighted antibiotic resistance as a global threat to public health. This document warns of the risk of life of the patient affected by an infectious disease hitherto treated with such antibiotics and, consequently, points to the need to adopt urgent measures in the search for alternatives. WHO recommends that studies be carried out for the development of new antibiotic substances, requiring public efforts, as well as society,

15/113 meaning that effective measures are taken to delay the process of bacteria resistance.

[0044] It is worth noting that no important type of antibiotic has been developed since the late 1980s, according to the WHO.

[0045] From 2011 to 2013, the FDA, the body that monitors the pharmaceutical and food industry in the USA, approved only three new molecular structures to fight bacterial diseases, the lowest number since the 1940s.

[0046] On the other hand, it is recognized that the resistance of microorganisms can also be promoted by the intensive use of biocides available on the market, with a lack of substitutes for these products that are compatible with the respective needs of different areas of application, whether in industry , the environment or health.

[0047] As can be seen in the state of the art discussed above, the proposals made do not fully meet the necessary requirements for safe Antimicrobial Compositions alternative to traditional commercial compositions. This situation remains a major challenge, as it has been a long-standing demand felt by the industry.

[0048] In view of the above, the state of the art does not describe or suggest the Antimicrobial Compositions of the present invention as described and claimed below, which are effective with a broad spectrum of action, but with minimal toxicity, for application in oil industry and in the health and environmental areas.

SUMMARY OF THE INVENTION

In a first aspect, the present invention consists of an Antimicrobial Composition containing a) 1% to 98% (weight / volume) of at least one bioactive compound selected from a compound of formula (I) and / or a compound of formula (IIA ) and / or a compound of formula (IIB):

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Formula (I) in which the respective substituents can be selected from the groups:

R-ι-ΟΗ e

R ₂ -H; OH; OCH ₃ and

R ₃ -H; OHe

R ₄ - H; OH and

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R ₅ - H; OH; OCH ₃

Re-H; OH; CH ₃ ; Cl; Br

R ₇ - H; OH; OCH ₃ ; NO ₂ ; SO ₃ H

Re-OH; OCH ₃ ; F; Cl; Br; CH ₃ ; NH ₂ ; NO ₂ ; SO ₃ H; C ₄ H ₉ NH ₂ ; OC ₂ H ₅ NH _2; OCH ₂ O

R ₉ -H; OH; OCH ₃ ; OCH ₂ O

R10-H; OH

Formula (IIA) Formula (IIB)

Rn-OH;

R1-H; OH;

b) acceptable vehicle; and c) optionally one or more adjuvants.

The Antimicrobial Composition of the invention, in addition to the broad spectrum of action against bacteria, is endowed with effectiveness and specificity in inhibiting the growth of different strains of bacteria that cause damage, respectively, to the oil industry, the environment and health.

The Antimicrobial Composition is particularly suitable for use in the prevention, mitigation and control of biogenic processes; in inhibiting the growth of strains of environmental bacteria and as a medicine.

[0049] In a second aspect, the present invention consists of a method for the prevention, mitigation and control of biogenic processes comprising the application of the composition described here in times of

18/113 adequate contact with the effective reduction of the contaminating population of planktonic or sessile bacteria and with isolated or intercropped action.

[0050] In a third aspect, the present invention consists of a method for inhibiting the growth of bacteria comprising the application of the composition described herein in contact times suitable for the effective reduction of the bacteria population of the contaminated environment to be treated.

[0051] In a fourth aspect, the present invention consists in the use of the composition described herein in the manufacture of a medicament for inhibiting bacterial targets.

[0052] In a fifth aspect, the present invention consists in the use of the composition described here in the prevention, mitigation and control of biogenic processes, preferably souring, corrosion influenced by microorganisms and buffering of oil wells, or inhibiting the degradation of corrosion inhibitors in diesel transportation pipelines.

[0053] In a sixth aspect, the present invention consists of the use of the composition described here in asepsis, disinfection and sterilization of domestic, hospital and industrial environments, including the prevention of contamination and reduction of infection in a hospital environment.

BRIEF DESCRIPTION OF THE FIGURES [0054] The detailed description presented below makes reference to the attached figures, which:

[0055] FIGURES 1 TO 3 are graphs representative of antimicrobial activity (biocidal efficiency), of particular compositions of the present invention, in challenge to different target cultures of BRS and compared to traditional commercial biocides GA (100 ppm) and THPS (100 ppm), comprising: [0056] Figures 1A, 1B and 1C - Floretine in concentrations of 150 and

300 ppm in DMSO and Floretina in concentrations of 150 and 300 ppm in synthetic seawater in defiance of Desulfovibrío alaskensis, BRS pool header P-19 and BRS pool T6 Lab, respectively;

[0057] Figures 2A, 2B and 2C - β-Lapachona and QUE-2, each at concentrations of 150 and 300 ppm in DMSO in defiance of, respectively, Desulfovibrío alaskensis, BRS pool header P-19 and BRS pool T6 Lab;

19/113 [0058] Figures 3A, 3B and 3C - Dihydrochalcone-1 dimer at concentrations of 500 and 1000 ppm in DMSO in defiance of Desulfovibrio alaskensis, BRS pool header P-19 and BRS ροοΙΎ6 Lab, respectively.

[0059] FIGURES 4 to 10 are graphs representative of the antimicrobial activity (biocidal efficiency) of particular compositions of the present invention in challenge to different target cultures of BRS, under different conditions and detailed in the examples, compared to traditional commercial biocides, comprising:

[0060] Figure 4 - QUE-2 and β-Lapachona, each in concentrations of 150 ppm and 300 ppm in DMSO in defiance of BRS pool header P-19, contact time of 18 hours, compared to THPS (100 ppm) ;

[0061] Figure 5 - β-Lapachona, at a concentration of 100 ppm in

DMSO, and Floretina, in a concentration of 50 ppm in DMSO in defiance of BRS pool header P-19 and Desulfovibrio alaskensis, contact time of 18 hours, compared to GA (100 ppm);

[0062] Figure 6 - QUE-2, β-Lapachona, and Floretina, each in concentration of 300 ppm, in DMSO, in challenge to BRS pool FPSO P-32, BRS pool FPSO P-43 and BRS pool header P- 19, contact time of 18 hours, compared to THPS (100 ppm);

[0063] Figure 7 - QUE-2 at a concentration of 300 ppm in DMSO (after dissolution in microcosm of deaerated synthetic seawater, it is filtered in a deaerated environment by NLO in Millipore membrane 0, 22 pm), in defiance of the target culture BRS pool header P-19, contact time of 18 hours, compared to THPS (100 ppm);

[0064] Figure 8 - QUE-2 at a concentration of 300 ppm in DMSO (after dissolution in microcosm of deaerated synthetic seawater, it is filtered in a deaerated environment by NLO in Millipore 0, 22 pm membrane) in defiance of the target cultures of BRS pool header P-19 and BRS pool FPSO P-32, in contact time of 4 hours and 24 hours, compared to THPS (100 ppm);

[0065] Figure 9 - QUE-2, in concentration of 300 ppm, in DMSO in defiance of BRS pool header P-19 and BANHT pool header P-19, contact times of 4 hours and 24 hours, compared to THPS (100 ppm);

20/113 [0066] Figure 10 - QUE-2 and β-Lapachona, each in concentrations of 100 ppm, 150 ppm and 300 ppm in DMSO and Floretina, in concentrations of 150 ppm and 300 ppm in DMSO in defiance of the BRS pool header P-19, contact times of 4 hours and 24 hours, compared to THPS (100 PPm).

[0067] FIGURES 11 (A and B) illustrate the control of microbial growth of planktonic and sessile BRS according to newly formed MES-biofilm, comprising:

[0068] Figure 11A - Representative graph of the antimicrobial activity (biocidal efficiency) of particular compositions of the present invention containing QUE-2, β-Lapachona, and Floretina, each in concentrations of 1000 ppm in defiance of the BRS pool header of P-19 in the process of adhering to SAE 1020 carbon steel biocoupons (newly formed biofilm) arranged in microcosm consisting of de-aerated sea water - (re-disclosure applied to MES) - in front of GA (500ppm) and THPS (500ppm);

[0069] Figure 11B - Representative graph of the antimicrobial activity (biocidal efficiency) of particular compositions of the present invention containing QUE-2, β-Lapachona, and Floretina, each in concentrations of 1000 ppm in defiance of the control culture Desulfovibrío alaskensis in process adherence to SAE 1020 carbon steel biocoupons (newly formed biofilm) arranged in a microcosm consisting of de-aerated sea water (disclosure applied to MES) - against GA (500 ppm) and THPS (500 ppm).

[0070] Figures 12 (A, B, C and D) are graphs that illustrate the evaluation of parameters associated with the corrosive process influenced by microorganisms (CIM) - MES (newly formed biofilm) by mass loss: particular compositions of the present invention - QUE-2, β-Lapachona, and Floretina, each in a concentration of 1000 ppm, traditional commercial biocides GA and THPS, both in the concentration of 500 ppm, and their respective chemical and biological controls, for two contact periods (15 and 45 days) in front of two BRS cultures and there is a classificatory character of degrees of risk, which is applied to the assessment of the aggressiveness of injection waters, in terms of generalized corrosion and localized corrosion, comprising:

21/113 [0071] Figure 12A - Representative graphs of the uniform Corrosion Rate (mm / year) evaluated by mass loss in defiance of the BRS pool header of P-19 against the referred compositions in a contact time of 15 days;

[0072] Figure 12B - Representative graphs of the Uniform Corrosion Rate (mm / year) evaluated by loss of mass in defiance of Desulfovibrio alaskensis compared to the mentioned compositions in a contact time of 15 days;

[0073] Figure 12C - Representative graphs of the uniform Corrosion Rate (mm / year) evaluated by loss of mass in defiance of the BRS pool header of P-19 compared to the referred compositions in contact time 45 days;

[0074] Figure 12D - Representative graphs of the uniform Corrosion Rate (mm / year) evaluated by loss of mass in defiance of Desulfovibrio alaskensis compared to the referred compositions in contact time 45 days.

[0075] Figures 13 (A and B) are graphs that illustrate the expectation of uniform corrosion protection, comprising:

[0076] Figure 13A - Expectation of uniform corrosion protection (biocoupons - SAE 1020 steel) against particular compositions of the present invention containing QUE-2, β-Lapachona, and Floretina (each at a concentration of 1000 ppm), as well as traditional commercial biocides GA and THPS (each at a concentration of 500 ppm) in the presence of the target culture BRS pool header P-19 in seawater (MES- contact 45 days);

[0077] Figure 13B - Expectation of uniform corrosion protection (biocoupons - SAE 1020 steel) against particular compositions of the present invention containing QUE-2, β-Lapachona, and Floretina (each at a concentration of 1000 ppm), as well as traditional commercial biocides GA and THPS (each at a concentration of 500 ppm) in the presence of the control culture Desulfovibrio alaskensis in seawater (MES- contact 45 days).

[0078] FIGURE 14 is a graph representing the Biocidal Efficiency of a particular composition of the present invention containing QUE-2 (1000 ppm) and the traditional commercial biocide THPS (500 ppm) in defiance of the BRS pool header of planktonic and sessile P-19 (preformed biofilm) (re-development applied to MES).

22/113 [0079] FIGURE 15 is a graph representing the Biocidal Efficiency of a particular composition of the present invention containing QUE-2 (1000 ppm) in challenge to planktonic and sessile BRS (biofilm-preformed) originating from pool header culture P-19.

[0080] FIGURES 16 (A, B, C, D, E, F and G) represent photomicrographs relating to the ultrastructural characterization by Transmission Electron Microscopy (MET) of the target culture Desulfovibrio alaskensis showing:

[0081] Figure 16A - CB1 (Biological Control 1) - target culture in deaerated sea water, with well preserved membrane and cytoplasm and dimensions of approximately 1.0 pm;

[0082] Figure 16B - CB2 (Biological Control 2) - target culture in

DMSO (300 ppm) in de-aerated sea water, with preserved morphology compared to the control situation CB1, and with characteristic dimension of BRS;

[0083] Figure 16C - Action of the commercial biocide GA (100 ppm) - target culture with disorganized external membrane (thick arrow) and slightly extracted cytoplasmic content, with a smaller dimension (~ 0.8 pm) compared to the control situation CB1 in the presence of depleted sea water;

[0084] Figure 16D - Action of the commercial biocide THPS (100 ppm) target culture with very disorganized external membrane (thick arrow) and slightly extracted cytoplasmic content, with a smaller dimension (~ 0.8 pm) compared to the control situation CB1 in presence of depleted sea water;

[0085] Figure 16E - Action of the composition containing Floretine (300 ppm) - target culture with well-preserved external membrane, showing, however, extensive cytoplasmic extraction (star) and smaller dimension (~ 0.8 pm) compared to the control situation CB2 in the presence of de-aerated sea water with DMSO;

[0086] Figure 16F - Action of the composition containing β-Lapachona (300 ppm) - target culture with external membrane detached from the internal membrane (thick arrow) and fragmented cytoplasmic content (star);

23/113 [0100] Figure 16G - Action of the composition containing QUE-2 (150 ppm)

- target culture with a membrane that has blebs (thin arrow) and highly extracted cytoplasmic content (star), indicating that the molecule permeates the membrane and internally triggers the bacterial death process.

[0101] FIGURES 17 (A, B, C, D, E, F and G) represent photomicrographs relating to the ultrastructural characterization by Transmission Electron Microscopy (MET) of the target culture BRS pool header P-19 showing:

[0102] Figure 17A - CB1 (Biological Control 1) - target culture in de-aerated seawater, with well preserved membrane and cytoplasm and characteristic BRS dimension (~ 1.0 pm to 1.5 pm);

[0103] Figure 17B - CB2 (Biological Control 2) - target culture in

DMSO (300 ppm) in de-aerated seawater, with a well-preserved membrane and intact cytoplasmic content, and with a characteristic BRS dimension (—1.0 pm to 1.5 pm);

[0104] Figure 17C - Action of the commercial biocide GA (100 ppm) - target culture with disorganized external membrane (thick arrow), with a smaller dimension (between 0.6 pm and 0.7 pm) compared to the control situation CB1 in the presence of depleted sea water;

[0105] Figure 17D - Action of the commercial biocide THPS (100 ppm) target culture with external membrane that detaches from the internal membrane (thick arrow) and slightly extracted cytoplasmic content, with a smaller dimension (—0.8 pm), compared to control situation CB1 in the presence of depleted sea water;

[0106] Figure 17E - Action of the composition containing Floretine (300 ppm) - target culture with well-preserved membrane, but showing small blebs (thin arrow), and with very extracted cytoplasmic content (star);

[0107] Figure 17F - Action of the composition containing β-Lapachona (300 ppm) - target culture showing altered membrane (thin arrow) and extracted cytoplasmic content (star), in addition to reduced size (~ 0, 6 pm) compared to control situation CB2 in the presence of de-aerated sea water plus DMSO;

24/113 [0108] Figure 17G - Action of the composition containing QUE-2 (150 ppm)

- target culture with a membrane with a large bleb (thin arrow) and very extracted cytoplasmic content (star).

[0109] Figures 18 (A, B, C, D and E) represent photomicrographs relating to the ultrastructural characterization by Transmission Electron Microscopy (MET) of the target culture BRS pool T6 Lab (antimicrobial test of example 1) showing:

[0110] Figure 18A - CB1 (Biological Control 1) - target culture in de-aerated seawater, with well preserved membrane and cytoplasm and characteristic BRS dimension (~ 1.0 pm to 1.5 pm);

[0111] Figure 18B - CB2 (Biological Control 2) - target culture in

DMSO (300 ppm) in de-aerated sea water, with a well-preserved membrane and intact cytoplasmic content, and with a characteristic BRS dimension (~ 1.0 pm to 1.5 pm) in de-aerated sea water;

[0112] Figure 18C - Action of the commercial biocide GA (100 ppm) - target culture with disorganized external membrane (thick arrow) and slightly extracted cytoplasmic content, and with a smaller dimension (—0.6 pm) compared to the control situation CB1 in presence de-aerated sea water;

[0113] Figure 18D - Action of the commercial biocide THPS (100 ppm) target culture with disorganized external membrane (thick arrow), condensed cytoplasmic content, with a smaller dimension (—0.6 pm) paralleling the control situation CB1 in the presence of deserted sea water;

[0114] Figure 18E - Action of the composition containing Floretine (300 ppm) - target culture with slightly wavy membrane and discreet presence of blebs (thin arrow), with extracted cytoplasmic content (star), and with reduced dimension (—0.5 pm) compared to the control situation CB2 in the presence of de-aerated sea water with DMSO.

[0115] Figures 19 to 22 represent photomicrographs related to the ultrastructural characterization by Scanning Electron Microscopy (SEM) of BRS pool header P-19 biofilms recently formed in carbonocarbon biocoupons, after being removed from the test solutions (contacts) a specified below:

25/113 [0116] Figure 19 - SEM photomicrograph of newly formed BRS pool header P-19 in carbon steel biocoupon, after 18 hours of contact in the presence of de-aerated sea water (control situation CB1).

[0117] Figure 20 - SEM photomicrograph of a newly formed BRS pool header P-19 in carbon steel biocoupon, after 18 hours of contact in the presence of de-aerated sea water plus DMSO (control situation CB2).

[0118] Figure 21 - SEM photomicrograph of newly formed BRS pool header P-19 in carbon steel biocoupon, after 18 hours in seawater in the presence of the commercial biocide THPS (500 ppm).

[0119] Figure 22 - SEM photomicrograph of newly formed BRS pool header P-19 in carbon steel biocoupon, after 18 hours of contact in seawater in the presence of the particular composition of the present invention containing QUE-2 ( 1000 ppm).

[0120] FIGURES 23 to 25 represent photomicrographs relating to the ultrastructural characterization by SEM of BRS pool header P-19 biofilms preformed in carbon steel biocoupons, after being removed from the test solutions (contacts) specified below:

[0121] Figure 23 - SEM photomicrograph of BRS pool header P-19 biofilms preformed in carbon steel biocoupon, after 24 hours of contact in the presence of de-aerated sea water plus DMSO (control situation CB2).

[0122] Figure 24 - SEM photomicrograph of BRS pool header P-19 preformed in carbon steel biocoupon, after 24 hours of contact in seawater in the presence of the traditional commercial biocide THPS (500 ppm).

[0123] Figure 25 - SEM photomicrograph of BRS pool header P-19 preformed in carbon steel biocoupon, after 24 hours of contact in seawater in the presence of the particular composition of the present invention containing QUE-2 ( 1000 ppm).

DETAILED DESCRIPTION OF THE INVENTION [0124] Preliminarily, it is emphasized that the following description will start from preferred embodiments of the invention. As will become evident to

26/113 any person skilled in the art, however, the invention is not limited to those particular embodiments.

[0125] Unless defined differently, all technical and scientific terms used herein have the same meaning as understood by a technician in the subject to which the present invention belongs. The terminology used in describing the invention is intended to describe particular embodiments only, and is not intended to limit the scope of its teachings.

[0126] Unless otherwise indicated, all numbers expressing quantities, percentages and proportions, and other numerical values used in the specification and in the claims, should be understood as being modified, in all cases, by the term “ about". Thus, unless otherwise indicated, the numerical parameters shown in the specification and in the claims are approximations that may vary depending on the properties to be obtained.

[0127] All patent or non-patent references cited in this application are hereby incorporated by reference in their entirety.

[0128] The Antimicrobial Compositions of the present invention constitute alternatives to commercially available biocides and / or antibiotics and / or their associations with other substances. The compositions described here have selective and specific action regarding the microbial target that is intended to be controlled and are effective in a short response time, minimizing risks to health and the environment.

[0129] In a non-limiting way, the compositions of the present invention can be applied in several industrial sectors, such as: improved recovery systems by water injection, especially sea water, and fresh water, re-produced oily water, oil reservoirs inshore and offshore, production systems, storage systems in terminals and / or transportation of oil (crude), natural gas and / or derivatives, vessel reservoirs and / or platforms that treat and store oil and / or oily water, systems gas washing, completion, stimulation, and drilling fluids applied to oil reservoirs, seawater and / or water based fluids

27/113 sweet for hibernation of pipelines in the commissioning phase (Hydrostatic Test), industrial lubricants, traditional fuels and / or biofuels, specific membrane filtration units, as in the case of removal of sulfate from seawater for injection into wells oil, and desalination of sea water for human use, paints and / or internal and external coatings of tanks and / or ships and / or platforms, cutting fluids, heat transfer systems, and other industrial and / or service segments intended for the preservation of different environments, such as: water collection and preservation systems for drinking and / or recreation, superficial and subsurface water sources, water-powered reactors, cooling systems in open or closed circuits, consisting of water sweet and / or salt water, industrial waters such as water for use in the manufacture of paper and cellulose, and food, systems sterilization and / or control of biological agents including home environments, public and private services, outpatient, laboratory, hospital, animal husbandry and / or food processing and also of vegetables, their packaging, including agribusiness, and their sanitizing uses, control of etiological agents with a focus on human and / or animal health and quality of life. The compositions described here can also be used in the pharmaceutical industry, being an important alternative to traditional antimicrobials for the treatment of bacterial infectious processes.

[0130] The Antimicrobial Composition of the present invention is comprised of (a) 1% to 98% (weight / volume) of at least one bioactive compound from the group of compounds selected from a compound of formulas (I) and / or (IIA) and / or (IIB):

Formula (I)

11/28

Formula (IIA)

Formula (IIB) in which the respective substituents can be selected from the groups:

Ri-OH and

R ₂ -H; OH; OCH ₃ and

R ₃ -H; OH and

R ₄ -H; OH and

R ₅ -H; OH; OCH ₃

11/29

or

R ₆ -H; OH; CH ₃ ; Cl; Br

R ₇ - H; OH; OCH ₃ ; NO ₂ ; SO ₃ H

Re-OH; OCH ₃ ; F; Cl; Br; CH ₃ ; NH ₂ ; NO ₂ ; SO ₃ H; C ₄ H ₉ NH ₂ ; OC ₂ H ₅ NH _2; OCH ₂ O

R ₉ -H; OH; OCH ₃ ; OCH ₂ O

Rio-H; OH

Rn-OH;

R1-H; OH;

b) acceptable vehicle; and c) optionally one or more adjuvants.

[0131] The compounds of formula (I) are composed of phenolic skeleton of the class of flavonoids (C6-C3-C6) and diflavonoids (C6-C3-C6) ₂ . Preferably, said compounds are in the category of chalcones, dihydrochalcones and dimeric dihydrochalcones. More preferably, said compounds are dihydrochalcones selected from 3- (4-hydroxyphenyl) -1- (2,4,6-trihydroxyphenyl) -1-propanone (Floretine) and 3- (4-methoxyphenyl) -1 - (2,4,6-trihydroxyphenyl) ) -1-propanone (QUE-2), dimeric dihydrochalcones such as 1.1 (2.2 ', 4.4', 6.6'-hexahydroxy [1,1 '-biphenyl] -3,3'-diyl ) bis [3- (4-methoxyphenyl) -1-propanone (dihydrochalcone dimer-1) and chalcones selected from 2 ', 3,4,4'-tetrahydroxychhalcona (butein) and 2', 4,4 ', 6'- tetrahydroxychalcona (chalcone naringenin).

[0132] The compounds of formulas (IIA) and (IIB) are phenolic skeletal compounds of the naphthoquinone class (C6-C4). Preferably, said compounds are 2-hydroxy-3- (3-methyl-2-butenyl) -1,4-naphthalenedione (Lapachol) and the compound of formula (IIB) is 2,2-dimethyl-3,4- dihydro-2Hbenzo [h] chromene-5,6-dione (β-Lapachona).

30/113 [0133] Table 2 illustrates the structure, nomenclature and registration number (CAS) reported in Chemical Abstracts of some preferred compounds of the present invention.

TABLE 2: Examples of preferred compounds of the invention

β-Lapachona (CAS 4707-32-8) (2H-naphtho [1,2-b] pyran-5,6-dione, 3,4dihydro-2,2-dimethyl)

Floretine (CAS 60-82-2) [3- (4-hydroxyphenyl) -1 - (2,4,6-trihydroxyphenyl) 1-propanone]

Dihydrochalcona-1 dimer

Dihydrochalcona-1 (QUE-2) (CAS 76172-68-4) [3- (4-methoxyphenyl) -1 -2,4,6trihydroxyphenyl) -1 -propanone] (CAS 198137-78-9) ( 1-propanone, 1, Γ- (2,2 ', 4,4', 6,6'hexahydroxy [1,1'-biphenyl] -3,3'-diyl) bis [3- (4 methoxyphenyl) [0134] In a first embodiment, the composition described here is used in the prevention, mitigation and control of biogenic processes.

[0135] When used for this purpose, at least one bioactive compound is comprised in concentrations in the range of 10 to 100,000 ppm, preferably from 10 to 5,000 ppm, more preferably from 50 to 1,000 ppm.

31/113 [0136] In a preferred embodiment, said at least one bioactive compound is selected from QUE-2, β-Lapachona, Floretina or dihydrochalcona-1 Dimero.

[0137] In a more preferred embodiment, the bioactive compound is QUE-2 comprised at a concentration in the range of 100 to 1,000 ppm, preferably about 300 ppm; or the bioactive compound is β-Lapachona comprised in a concentration in the range of 100 to 1,000 ppm; or the bioactive compound is Floretine comprised in a concentration in the range of 50 to 300 ppm, preferably from 150 to 300 ppm; or the bioactive compound is Dimero dihydrochalcona-1 comprised in a concentration in the range of 150 to 1,000 ppm.

[0138] In a preferred embodiment, said acceptable vehicle comprises one or more vehicles selected from DMSO; sea water; fresh water; brackish water; isopropanol; glycerol and / or other higher alcohols, such as propylene glycol; sodium chloride and / or other salts.

[0139] In a second embodiment, the composition described herein is used to inhibit the growth of strains of environmental bacteria.

[0140] When used for this purpose, at least one bioactive compound is comprised in concentrations in the range of 1 to 10,000 ppm.

[0141] In a preferred embodiment, said at least one bioactive compound is selected from QUE-2, β-Lapachona, Floretina or dihydrochalcona-1 Dimero.

[0142] In a more preferred embodiment, the bioactive compound is QUE-2 comprised in a concentration in the range of 1 to 50 ppm, preferably from 1 to 30 ppm, more preferably from 5 to 30 ppm, even more preferably from 10 to 25 ppm; or the bioactive compound is β-Lapachona comprised in a concentration in the range of 1 to 50 ppm, preferably from 1 to 30 ppm, more preferably from 1 to 15 ppm; or the bioactive compound is Floretine comprised in a concentration in the range of 1 to 50 ppm, preferably from 1 to 30 ppm.

[0143] In a preferred embodiment, said acceptable vehicle comprises one or more vehicles selected from DMSO; natural water

32/113 low salinity; sea water; fresh water; glycerol and / or other alcohols such as ethanol, propanol or isopropanol; higher alcohols such as isoamyl or propylene glycol; sodium chloride and / or other salts.

[0144] In a third embodiment, the composition described herein is used as a medicine.

[0145] When used for this purpose, at least one bioactive compound is comprised in concentrations in the range of 1 to 8,000 ppm.

[0146] In a preferred embodiment, said at least one bioactive compound is selected from QUE-2, β-Lapachona, Floretina or Dihydrochalcona-1 dimer.

[0147] In another more preferred embodiment, the bioactive compound is QUE-2 comprised in a concentration in the range of 15 to 1500 ppm, preferably from 20 to 700 ppm, even more preferably from 15 to 100 ppm; or the bioactive compound is β-Lapachona comprised in a concentration in the range of 50 to 1,500 ppm, preferably from 80 to 1,300 ppm, even more preferably from 70 to 100 ppm.

[0148] In another more preferred embodiment, the bioactive compound is QUE-2 comprised in a concentration in the range of 1 to 50 ppm, preferably from 5 to 30 ppm; even more preferably from 10 to 25 ppm; or the bioactive compound is β-Lapachona comprised in a concentration in the range of 1 to 50 ppm, preferably from 1 to 30 ppm, still more preferably from 1 to 15 ppm; or the bioactive compound is Floretine comprised in a concentration in the range of 1 to 50 ppm, preferably from 1 to 30 ppm.

[0149] In a preferred embodiment, said acceptable vehicle comprises one or more vehicles selected from DMSO, lactose, stearate, starch, carbonates and / or, bicarbonates, methylcellulose, sodium alginate, ethanol, glycerol, sorbitol, sterile distilled water, sterile saline, osmolarity buffers, pH and / or phosphate; zinc oxide and / or cellulose gum.

[0150] In one embodiment, the adjuvant is selected from chelating and surfactant agents, corrosion inhibiting substances, solid supports,

33/113 carriers of active substances, membrane permeabilizers, preservatives, and / or metabolic BRS inhibitors.

[0151] In a preferred embodiment, the chelating and surfactant agents are ethylenediamino tetraacetic acid (EDTA), ethylene diaminodisuccinate (EDDS) and molecular structures of the chemical category of saponins.

[0152] In a preferred embodiment, the corrosion inhibiting substances are selected from capsaicin and its analogs.

[0153] In a preferred embodiment, the solid supports are selected from silica, polymers derived from silica and polypropylene.

[0154] In a preferred embodiment, the carriers of the active substances are selected from cyclodextrins and derivatives.

[0155] In a preferred embodiment, metabolic inhibitors of

BRS are selected from selenite, selenate, monofluorophosphate, molybdate, nitrite, nitrate and their associations.

[0156] In one embodiment, said compositions are used in a method for the prevention, mitigation and control of biogenic processes, in which the compositions of the present invention are applied in contact times suitable for the effective reduction of the contaminating population of bacteria, which they can be planktonic or sessile and with isolated or intercropped action.

[0157] Preferably, said adequate contact time is about 4 to about 24 hours for planktonic bacteria and about 15 days for sessile bacteria.

[0158] Preferably, said bacteria are heterotrophic, mesophilic, aerobic, facultative or anaerobic, Gramnegative or Gram-positive, non-pathogenic or pathogenic bacteria.

[0159] In a preferred embodiment, said bacteria are sulfate-reducing bacteria forming biofilms originating from different sites in the oil industry, both surface and subsurface, of process and storage vessels for oil and gas, of collected fluid in an oil production system subject to souring and other inshore and offshore oil installations, and / or BANHT consortium with BRS.

34/113 [0160] This method promotes the prevention, mitigation and control of biogenic processes associated with said anaerobic organisms. In a non-limiting way, biogenic processes can be the acidification (souring) of oil reservoirs by the unwanted biogenic production of H2S, Corrosion Influenced by Microorganisms (CIM) and the plugging of oil wells, which are usually associated the presence of biofilms, generally related to FeS deposition.

[0161] In particular, the composition is used to prevent or significantly reduce the process of adhesion of BRS to carbon steel metallic surfaces, a previous step necessary for the formation of biofilms, controlling the growth of these bacteria in sessile form, and also acting preventively in the control of biogenic phenomena. Just as they act in the control of generalized corrosion, when the biofilm is preformed.

[0162] In another embodiment, the compositions of the present invention are used in a method for inhibiting the growth of bacteria, in which the compositions are applied at contact times suitable for effectively reducing the population of bacteria in the contaminated environment to be treated.

[0163] Preferably, said adequate contact time is 24 hours.

[0164] In the context of the present invention, the compositions described herein are effective over a broad spectrum of strains of environmental bacteria.

[0165] In a preferred embodiment, said bacteria are characterized as aerobic Gram-positive bacteria including some found in petroleum reservoirs located in deep sea waters.

[0166] In a more preferred embodiment, said bacteria are of the genus Bacillus, preferably Bacillus licheniformis, Bacillus amyloliquefasciens, Bacillus subtilis, Bacillus cereus, or Paenibacillus macerans.

35/113 [0167] The action of the compositions described here against the

Bacillus cereus due to the ability of this microorganism to degrade aliphatic and aromatic hydrocarbons present in corrosion inhibitors influencing corrosion in diesel transport ducts.

[0168] It was found that the compositions of the present invention have efficacy and specificity at low concentrations (Minimum Bactericidal Concentration - CMB), significant when seeking to reduce toxicity, highlighting the fact that the said microorganisms already show resistance to traditional antimicrobials.

[0169] The use of lower mass amounts of the composition to be used in the environment is of great benefit when seeking to reduce toxicity and alternatives that act to attenuate the antimicrobial resistance.

[0170] Thus, in one embodiment, the compositions described here are intended for asepsis, disinfection and sterilization of different environments, and can be used, for example, in the prevention of contamination and in the reduction of hospital infection and in the decontamination of environments in general, including domestic, industrial and services.

[0171] In another embodiment, the compositions of the present invention act in inhibiting pathogens as an alternative to antibiotics, and also in challenging bacteria of the genus Bacillus with impact on health and / or other bacterial targets of medicinally significant interest for human health and animal, for example, Enterococcus faecalis and Staphylococcus aureus.

[0172] The compositions of the present invention are still effective against a broad spectrum of bacteria of health interest comprising pathogenic strains, such as: S. aureus; E. coli; E. faecalis; P. aeruginosa; S. Enteritidis; S. Typhi; S. flexneri. Noteworthy is the performance at reduced concentration levels compatible with antibiotic compositions, against Enterococcus faecalis and Staphylococcus aureus.

[0173] Bacteria of the genus Bacillus of medicinally significant interest were also susceptible to the compositions of the present invention. Emphasis is given to the bacterium Bacillus cereus that causes food poisoning similar to that caused by Staphylococcus, which is a bacterium

36/113 admittedly pathogenic [Ryan K.J., Ray C.G. (editors). Sherris Medical Microbiology (4th ed.). McGraw Hill.ISBN 0-8385-8529-9. 2004], [0174] The significant effectiveness of the compositions of the invention against Gram-positive pathogens known to be resistant to antibiotics, such as S. aureus and E. faecalis, is also emphasized in reduced CMB values representative of antibiotic activity, as well as their efficacy against strains of Gram-negative health bacteria at higher CMB values.

[0175] The following examples show bioactivity, expressed in

CMB of the composition containing a bioactive compound as described herein, demonstrating reduced values indicative of antibiotic activity against bacterial pathogens, including all bacteria of the genus Bacillus of medicinally significant interest.

[0176] Particularly, the composition containing the bioactive compound

QUE-2 shows reduced values of CMB at levels compatible with antibiotic compositions acting, respectively, against Enterococcus faecalis (80.32 ppm) and Staphylococcus aureus (20.08 ppm). Likewise, the composition containing the bioactive compound Floretina acts against Enterococcus faecalis (6.0 ppm) and Staphylococcus aureus (12.0 ppm), as well as, similarly, the composition containing the compound β-Lapachona against Enterococcus faecalis (80, 78 ppm).

[0177] Thus, in another embodiment, the compositions described herein are used for the manufacture of a medicament for inhibiting bacterial targets. Preferably, said bacterial targets are S. aureus, E. coli, E. faecalis, P. aeruginosa, S. Enteritidis, S. Typhi, S. flexneri, or Paenibacillus macerans or species of the genus Bacillus, preferably Bacillus licheniformis, Bacillus amyloliquefasciens , Bacillus subtilis, or Bacillus cereus.

[0178] In the context of the present invention, casts of target bacteria

Gram-positive and Gram-negative, mesophilic, aerobic and anaerobic, were tested, under conditions of different protocols, and in terms of their susceptibility to the active substances of the said compositions of the present invention.

37/113 [0179] The first consisted of the interest group of the oil industry, classified as anaerobic Gram-negative organisms, of the BRS, including four cultures of different origins (injection systems, production, oil and rock separation units) deep reservoir, in addition to a reference-control culture Desulfovibrio alaskensis, planktonic and sessile), as well as anaerobic bacteria (BANHT) intercropped with BRS, assessing abiotic and biotic corrosivity.

[0180] The second group of target bacteria consisted of 14 (fourteen) certified standard strains, Gram-positive and Gram-negative, of health bacteria and environmental bacteria, originating from Fiocruz reference banks. Among the strains of bacteria in the environmental area are two found in the deep reservoir rock.

[0181] The results obtained by the different applied techniques, made compatible, as shown in the following examples, allowed to select and attest, the antimicrobial activity of the substances that compose the compositions described here, with expected specificity in relation to the targets to be controlled. These results are therefore certified, as well as making it possible to prove the activity of the compositions of the present invention in preventing corrosion and inhibiting the formation of biofilms.

[0182] The compositions of the present invention are endowed with a wide potential spectrum for application in the health area, causing less environmental impact and / or being the object of easier deactivation, when compared to traditional commercial biocidal substances, as a rule classified as pesticides.

[0183] The compositions of the present invention can also be used in systems that apply alternatives of new technologies, such as those of seawater injection for improved recovery of oil equipped with membrane sulfate removal units (SRU), coupling dosage of metabolic inhibitors specific to BRS such as molybdate ion. Thus, it acts with the desired specificity, both in the viability of the biocidal action itself on BRS through the compositions of the present invention, as well as in blocking the sulfate-reducing metabolism of

38/113 referred bacteria, already in favorable stoichiometric conditions in the sulfate / molybdate ratio, which should minimize the concentration of H ₂ S in the fluids. [0184] The compositions of the present invention have the ability to act effectively, specifically and safely in the inhibition of BRS and, consequently, in the elimination of harmful effects resulting from the presence of these target microorganisms, as they meet the following technical requirements in the evaluations a that was submitted:

- Bactericidal efficiency of 99.90% and, or 99.99%, or higher, respectively, with minimum reduction in the concentration of the BRS bacterial population (planktonic and, or sessile) on the decimal logarithmic scale between three (3) and, or four (4) units and, or more than these; and / or total death understood as the non-detection of the target microorganism tested according to the applied technique;

Chemical compatibility according to the characteristics of the fluid to be treated (such as, for example, fresh water, sea water, oily water produced);

Stability in application and storage.

[0185] The compositions of the present invention also have the following particularities:

Antimicrobial bioactivity against BRS, both surface and subsurface, present in different scenarios in the oil industry, and in the environment;

- Inhibition of the generation of H ₂ S, the main metabolite of the sulfate reduction process, which is recognized as a highly corrosive substance and associated with souring, toxic character and water and air pollutant;

- Antioxidant characteristics useful in corrosion control;

- CIM inhibition, with minimal risk of corrosivity to carbon steel in the presence of de-aerated sea water.

[0186] The technical advantages with the application of the composition of the present invention particularly in the oil industry are:

Specificity of biocidal action against BRS to control microbiological corrosion and souring-,

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- Ability to prevent the formation and disorganization of bacterial biofilms;

- Broad spectrum to control other relevant groups of strains of environmental bacteria and health bacteria

- .Efficiency against each group of bacteria to be controlled, with a minimum reduction of three to four units on the logarithmic scale in growth;

- Minimal risk of corrosivity to carbon steel (abiotic and biotic) in the presence of de-aerated sea water;

- Chemical compatibility with fluids (especially fresh water and de-aerated seawater) and dosed products, such as oxygen scavengers;

- Potential for application in different scenarios of the oil industry;

- Potential for application in different scenarios in the areas of health and environment;

- Characteristic friendly to the safety and health of the worker and the environment; and

- Easy handling and stability in storage and application.

[0187] In summary, the compositions of the present invention are safe when compared to traditionally known commercial biocides, thus guaranteeing conditions of use compatible with the work safety requirements in industrial units.

EXAMPLES [0188] The following examples are illustrative and are intended to demonstrate the specific properties (biocidal performance and other characteristics) of particular compositions of the present invention, comprising preferred bioactive compounds and cannot, therefore, serve as a basis for restricting the scope of the present invention. .

[0189] The examples described below comprise tests demonstrating Biocidal Efficiency compared to the Antimicrobial Compositions of the present invention, in relation to target organisms in the oil industry, focusing on planktonic and sessile BRS cultures, including MIC control, as well as for relevant groups of strains of environmental bacteria and health bacteria.

40/113 [0190] The evaluation of the Biocidal Efficiency of the compositions of the present invention against target cultures of BRS, originating in the oil industry, was performed using the Most Likely Number (NMP) technique, according to protocols established from Postgate (1984 ), against these compositions and their respective biological controls, with certification tests in the presence of traditional commercial biocides glutaraldehyde (GA) and / or tetrakis (hydroxymethyl) phosphonium (THPS) sulfate. Optionally simulate the natural conditions of the target microorganisms, such as the typically anaerobic and saline environment, which is characteristic, for example, of overwintering fluids for Hydrostatic Testing and also seawater injection systems for recovery improved oil, especially at flow points downstream of the deaeration towers of such systems.

[0191] The Antimicrobial Compositions of the present invention, evaluated in the examples, comprise bioactive substances in concentrations close to the recommended range (10 mg / mL to 20 mg / mL) for laboratory tests and compatible vehicle such as, for example, dimethyl sulfoxide (DMSO ), from Sigma or equivalent, according to sterile conditions by filtration in Millipore membrane (0.22 pm), with preservation carried out in an atmosphere of oxygen-free nitrogen (NLO), in the absence of light, and under refrigeration. The DMSO vehicle was considered suitable for use after previous evaluation in terms of inhibitory concentration against the cultures BRS pool P-19 and Bacillus mycoides, a species previously isolated in consortium with BRS present at a sampling point close to the header of the referred system. injection of sea water. Alternatively, absolute ethanol, of Sigma origin or equivalent, and synthetic seawater itself (from Standard Methods) are applied as a vehicle for the direct dissolution of bioactive substances that are components of the compositions of the invention.

[0192] The BRS target cultures, characterized as Gram-negative bacteria, in planktonic and / or sessile forms, present themselves as typical organisms of the oil industry responsible for the generation of biogenic processes that are difficult to control, and are presented below:

Culture BRS pool header P-19, which originates from an injection system

41/113 sea water from Platform P-19, from Campo de Marlim, from Petrobras. BANHT pool cultures were detected in association with this BRS culture. Said BRS culture constitutes a test standard;

- Culture BRS Desulfovibrio alaskensis, which was isolated and identified in a study (FEIO, MJ et al. Desulfovibrio alaskensis sp. Nov., A sulphatereducing bacterium from a soured oil reservoir. International Joumal of Systematic and Evolutionary Microbiology, 54 (Pt 5) : 1747-52 September 2004) conducted in fluid collected in an oil production system (USA) subject to the occurrence of souring, belonging to the collection of strains of the University of Portsmouth (UK), being made available to Fiocruz and applied as a control culture ;

- Culture BRS pool T6 Lab, which originates from a core of deep oil rock from a well in the Campo de Marlim, Campos Basin, of Petrobras, being considered an indigenous culture, typical of a subsurface environment (SEBASTIÁN, G.V., 1999);

- BRS pool P-32 and BRS pool P-43 cultures, which originate from the respective Petrobras' FPSO (Floating Production Storage and Offloading) units in the Campos Basin, which process hydrocarbons, proceeding to their separation and treatment oily water produced for disposal according to release standards permitted by environmental agencies. In this system, in the tanks of these units, the constant presence of H2S, a typical BRS metabolite, is monitored and controlled.

[0193] The target cultures tested are maintained in the laboratory, according to a standardized protocol, by successive raising techniques in modified Postgate C medium, under anaerobic and mesophilic conditions (30 ° C), with monitored growth (range 10 ⁶ to 10 ⁸ cells / mL) by incubation in modified Postgate E medium. For the quantification of cell density (viability), the Most Likely Number (NMP) technique, developed from Postgate (1984) for desulfovibrios, is applied, with prior successive decimal dilutions (up to 10 ' ⁸ to 10' ⁹ ) in reducing saline solution (SSR), in sets of triplicates, after vortexing, for a period of about 30 seconds. The concentration of the BRS target culture, expressed as

42/113 viable bacterial cells, is determined as NMP per ml_, after consulting the table (W.F.Harringan), according to 95% confidence limits. Analogous quantification procedure is performed for target cultures of BANHT, in this case, applying specific culture medium for growth. The salinity of the culture media and other solutions is equivalent to that of the sea.

[0194] In summary, the antimicrobial assay targeting planktonic BRS consists of the contact step of the composition, added at the defined concentration, in the presence of a microcosm consisting of synthetic seawater (Standard Methods), and the resazurin oxirreduction indicator (0, 04% w / v), which is previously subjected to de-aeration by stripping with oxygen-free nitrogen (NLO). Defined aliquot (10% v / v) of BRS target culture, with a culture age of 18 hours in modified Postgate C medium, is inoculated, immediately, confirming the concentration (time = zero) in the contact in the range of 10 ⁶ - 10 ⁸ NMP / fnL, followed by hibernation for a defined period, generally of 18 hours and / or another reported, under anaerobic conditions, at 30 ° C. Biological controls in the presence of the bacterium and each of the constituents of the microcosm (de-aerated sea water, and de-aerated sea water plus an equivalent volume of the DMSO or other vehicle reported), as well as chemical controls (sterile), in the absence of the bacteria, are rehearsed. The tests and their respective controls are performed in triplicate, being pooled after the defined period of contact and, immediately, the BRS growth evaluation is carried out by incubation in modified Postgate E medium, according to the NMP technique, comparing the tests in the presence of the compositions to their respective biological controls, defining the Biocidal Efficiency. All tests are certified against traditional commercial biocides THPS and / or GA, in the concentration ranges referenced in the specialized literature for application in the oil industry. All steps of the procedure are performed under sterile conditions, in a vertical laminar flow chamber, and under NLO flow. The evaluation of the concentration of the BRS target culture is carried out, as previously reported, using the NMP technique (three tubes), the inoculation being done in decimal dilutions in modified Postgate E medium (from Postgate, 1984), with incubation in anaerobic conditions at a temperature of 30 ° C, and observation for a period of up to 28 days. Efficiency

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Biocide compared to the compositions of the invention and traditional commercial biocides is determined, after the fixed contact period, by the decrease in concentration (NMP / mL) relative to the respective microcosm in the presence of the vehicle in terms of its volumetric equivalent in seawater (Control Biological CB2, in the case of DMSO or other mentioned), being desirable to present in the order of 10 ³ to 10 ⁴ or higher, being represented as a percentage. Upon reaching a population reduction of the order of 10 ⁶ , it can be admitted that a sterilizing character (sterility assurance took (SAL) of 1CT ⁶ ) was achieved [von WOEDTKE et al. “The limits of sterility assurance”. GMS Krankenhhyg Interdiszip. 3 (3): Doc19. 2008] reported under the conditions applied to the test.

EXAMPLE 1 [0195] Antimicrobial compositions related, respectively, to the synthesized bioactive substances β-Lapachona, Floretina and QUE-2, using DMSO as the vehicle, were dosed in contact microcosm consisting of de-aerated sea water, in concentrations of 150 ppm and 300 ppm and the compositions related to the dihydrochalcona-1 dimer in concentrations of 500 ppm and 1000 ppm in DMSO with target in cultures of planktonic BRS pool header P-19, Desulfovibrio alaskensis and ροοΙΎδ Lab, in contact time of 18 hours. Compositions containing Floretine and synthetic sea water as a vehicle, in concentrations of 150 ppm and 300 ppm were also evaluated against the referred BRS cultures. It should also be noted that compositions containing this substance as an active compound, preserved for periods of about two years according to the procedure already reported, were also subjected to the test. The results of the tests to evaluate the Biocidal Efficiency of said compositions of the invention are presented in Tables 3 to 5 in sequence.

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TABLE 3: Results of tests to evaluate the Biocidal Efficiency of compositions containing Floretine, in concentrations of 150 ppm and 300 ppm in

DMSO and in concentrations of 150 ppm and 300 ppm in synthetic seawater

Planktonic BRS BRS target culture pool header P-19 BRS target culture Alaskensis desulfovibrio Target culture BRS pool T6 Lab Biological Controls BRS concentration (NMP / mL) Culture, dil 10 ¹ inoculated 1.4x10 ' 4.5 x 10 " 1.4 x 10 " CB1 - sea water 4.5 x 10 " 1.1 x 10 " 1.1 x 10 " CB2 - sea water + DMSO 150 ppm 1.4 x 10 " 1.4 x 10 " 1.4 x 10 " CB2 - sea water + DMSO 300 ppm 1.1 x 10 " 1.5 x 10 " 4.5 x 10 ⁴ Compositions BRS concentration (NMP / mL) Antimicrobials (Biocidal efficiency%) Floretine 150 ppm in <0.3 <0.3 4.5 x 10 sea water * (100%) (100%) (99.96%) Floretine 300 ppm in <0.3 <0.3 0.9 X 10 sea water * (100%) (100%) (99.99%) Floretine 150 ppm in <0.3 <0.3 <0.3 DMSO (100%) (100%) (100%) Floretine 300 ppm in <0.3 <0.3 <0.3 DMSO (100%) (100%) (100%) Floretine 150 ppm in <0.3 <0.3 <0.3 DMSO * (100%) (100%) (100%) Floretine 300 ppm in <0.3 <0.3 <0.3 DMSO * (100%) (100%) (100%) GA 100 ppm certification <0.3 <0.3 <0.3 (100%) (100%) (100%) THPS 100 certification <0.3 <0.3 <0.3 PPm (100%) (100%) (100%) ‘Composition containing Floretine Ensaiac the after preservation time of two

years

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TABLE 4: Results of tests to evaluate the Biocidal Efficiency of compositions containing QUE-2, β-Lapachona in concentrations of 150 ppm and

300 ppm in DMSO

Planktonic BRS Target culture BRS pool header P-19 Target culture BRS Desulfovibrio alaskensis Target culture BRS pool T6 Lab Biological Controls BRS concentration (NMP / mL) Inoculated dil 10 ' ¹ culture 1.4 x 10 ⁸ 1.1 x 10 ⁶ 1.1 x 10 ⁷ CB1 - sea water 4.5 x 10 ⁵ 1.1 x 10 ⁵ 1.4 x 10 ⁵ CB2 - sea water + DMSO 150 ppm * 2.5 x 10 ⁵ 1.4 x 10 ⁵ 1.1 x 10 ⁵ CB2 - sea water + DMSO 300 ppm * 4.5 x 10 ⁵ 1.4 x 10 ⁵ 1.1 x 10 ⁵ Compositions Antimicrobials BRS concentration (NMP / mL) (Biocidal efficiency%) QUE-2 150 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) QUE-2 300 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) β-Lapachona 150 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) β-Lapachona PS 300 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) GA 100 ppm certification <0.3 (100%) <0.3 (100%) <0.3 (100%) THPS 100 certification PPm <0.3 (100%) <0.3 (100%) <0.3 (100%)

* CB2 common to the two compositions tested, respecting the applied concentrations

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TABLE 5: Results of tests to evaluate the Biocidal Efficiency of compositions containing Dihydrochalcona-1 Dimer in concentrations of 500 ppm and 1000 ppm in DMSO.

Planktonic BRS Target culture BRS pool header P-19 BRS target culture Alaskensis desulfovibrio Target culture BRS pool T6 Lab Biological Controls BRS concentration (NMP / mL) -1 Culture, inoculated dil 10 1.4 x 10 ⁷ 1.1 x 10 ⁶ 4.5 x 10 ⁶ CB1 sea water 1.4 x 10 ⁶ 3.0 x 10 ⁵ 4.5 x 10 ⁴ CB2 sea water + DMSO 500 ppm 4.5 x 10 ⁵ 2.5 x 10 ⁵ 4.5 x 10 ³ CB2 sea water + 1000 ppm DMSO 4.5 x 10 ⁵ 1.4 x 10 ⁵ 1.4 x 10 ⁴ Compositions Antimicrobials BRS concentration (NMP / mL) (Biocidal efficiency%) Dimer dihydrochalcona-1 500 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) Dimer dihydrochalcona-1 1000 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) THPS 50 ppm certified <0.3 (100%) <0.3 (100%) <0.3 (100%) GA 50 ppm certification <0.3 (100%) <0.3 (100%) <0.3 (100%)

[0196] The results demonstrate the antimicrobial bioactivity of the composition containing Floretine in synthetic seawater in the concentration range 150-300 ppm, with the Biocidal Efficiency of 100% in the assay against cultures of BRS Desulfovibrio alaskensis and pool header P-19 and with Biocidal efficiency above 99.96% when tested against culture of BRS pool T6 Lab. Also the composition containing Floretine, in the concentration range 150-300

47/113 ppm in DMSO, presents antimicrobial bioactivity with Biocidal Efficiency of 100% in the assay against the three BRS cultures tested Desulfovibrio alaskensis, pool header P-19 and pool T6 Lab. It is worth mentioning the fact that the composition containing the substance Floretina maintained antimicrobial bioactivity even two years after its preparation. This translates into a significant indication of the half-life of said substance, that is, its stability, attesting to the preservation defined in the test protocol.

[0197] Relevant is the evidence that compositions previously described against the target cultures of BRS demonstrated Biocidal Efficiency of 100% in concentrations in the range of 150-300 ppm (β-Lapachona, Floretina, QUE-2) and in concentrations in the range 500-1000 ppm (Dihydrochalcona-1 dimer), all in the presence of the DMSO vehicle. There was also a significant Biocidal Efficiency (99.93%) against target culture pool header P-19, in planktonic form, which was previously evidenced by the composition containing the substance Dihidrochalcona-1 dimer, in the concentration of 172 ppm in the presence of the DMSO vehicle. This composition was exceptionally diluted directly in the microcosm of de-aerated seawater, in the said concentration, in the presence of a more conservative inoculum (10 ⁷ NMP / mL) of the target culture BRS (respective Biological Control), in the proportion of 20% v / v , featuring biocidal activity against BRS, in the usual range of antibiotics, as shown in Table 6 below.

TABLE 6: Test result for evaluating the Biocidal Efficiency of compositions containing Dihydrochalcone -1 dimer at a concentration of 172 ppm in DMSO.

Antimicrobial Composition Target culture BRS planktonic pool header P-19 CB2 Biological Control Concentration (NMP / mL) Composition * Concentration (NMP / mL) (Biocidal Efficiency%) * Dihydrochalcone dimer - 1 172 ppm * 1.0 x 10 ⁷ 0.7 x 10 ³ (99.93%)

48/113 [0198] The results presented in Tables 3 to 5 were certified by the biocides GA and THPS, in concentration in the range 50 -100 ppm, with the Biocidal Efficiency range of 100%.

[0199] These results are represented graphically, showing the antimicrobial bioactivity against the cultures of BRS Desulfovibrio alaskensis, pool header P-19 and pool T6 Lab, respectively, for each of the compositions: Floretine in synthetic seawater and in DMSO ( Figures 1A, 1B and 1C), β-Lapachona and QUE-2 each in DMSO (Figures 2A, 2B and 2C) and Dihydrochalcone-1 dimer in DMSO (Figures 3A, 3B and 3C), compared to traditional commercial biocides GA and THPS.

[0200] Ultrastructural analyzes were performed by Microscopy

Transmission Electronics (MET) of the target cultures of BRS, in the planktonic form Desulfovibrio alaskensis, pool header P-19 and pool T6 Lab, compared to the compositions of the invention containing QUE-2, Floretine and β-Lapachona and traditional commercial biocides GA and THPS, as well as the corresponding biological controls, in which such compositions showed growth inhibition by the NMP technique, in order to verify possible structural changes and evaluate the mechanism of cell death.

[0201] the methodology for processing the bacterial samples used fixation for 24 h at 4 ° C, in 2.5% w / v glutaraldehyde in 0.1 M cacodylate buffer, pH 7.6. The samples were washed in the same buffer, for 3 times for 20 min each time, post-fixed in OsO ₄ 1% w / v solution at room temperature for 1.5 h, in the dark and washed in buffer. To preserve extracellular polymeric substances (SPE), ruthenium red (VR), a marker for anionic glycosylated polymeric substance, the buffers were added in a concentration of 0.14% w / v during the fixation and post-fixation steps and 0.07% w / v during the washing steps. The samples were dehydrated in increasing concentrations of acetone (30%, 70%, 80%, 90% and 100% acetone - 2 times of 15 min). The samples were included in Epon resin and, after 72 hours of polymerization, the material was sectioned (ultrathin sections) in a Leica ultramicrotome. The grids were contrasted in uranyl acetate and lead citrate and then

49/113 the Zeiss EM10C transmission electron microscope was observed.

[0202] The results of the analysis of Electron Microscopy of

Transmission (MET), referring to the three (3) cultures of BRS Desulfovibrio alaskensis (Figures 16 A, B, C, D, E, F, G), pool header P-19 (Figures 17 A, B, C, D, E, F, G) and T 6 Lab pool (Figures 18 A, B, C, D, E), in planktonic form, tested against the compositions of the invention containing QUE-2, Floretina and β-Lapachona, and biocides traditional commercial GA and THPS, when compared to their respective Biological Controls, demonstrate that the morphological changes observed express full compatibility with the high Biocidal Efficiency and specificity achieved, as described in example 1. In addition, they prove the absence of DMSO interference in antimicrobial activity of these compositions by showing that it does not cause significant ultrastructural changes in the target cultures.

[0203] Figures 16 and 17 indicate that the molecule permeates the membrane and internally triggers the bacterial death process.

EXAMPLE 2 [0204] Evaluation of the antimicrobial activity of compositions containing QUE-2 and β-Lapachona, in concentrations of 150 ppm and 300 ppm in DMSO, in a microcosm consisting of deaerated synthetic seawater, against the target culture of BRS pool header P-19, in planktonic form, with the purpose of assessing whether these compositions meet the needs of the oil industry for effective biocidal action in lower concentrations, compared to Biological Controls (CB2), in challenge to the target culture in higher concentrations elevated, in the order of 10 ⁶ NMP / mL, and in contact time of 18 hours.

[0205] The results observed in Table 7 below demonstrate the achievement of high degrees of Biocidal Efficiency, indicating values equal to or greater than 99.99%, compared to the composition containing β-Lapachona, and 100% or total death (no detection using the NMP technique) compared to the composition containing QUE-2 (Figure 4). The certification was carried out against the traditional commercial biocide THPS at a concentration of 100 ppm.

50/113 [0206] It should be noted that, in this test, the susceptibility of that culture was demonstrated, in high bacterial concentration, compared to both compositions also in the lowest concentration of these tested, of 150 ppm, with emphasis on the reach of total death from the composition containing QUE-2, which characterizes the potential for sterilization in contact time of 18 hours with a target in the referred planktonic BRS culture tested.

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TABLE 7: Results of the evaluation of the antimicrobial activity of the compositions comprising QUE-2, β-Lapachona, in concentrations of 150 ppm and 300 ppm in DMSO.

Planktonic BRS BRS pool target culture header P-19 Biological Controls Concentration (NMP / mL) Inoculated control culture (without dilution) 1.4x 10 ⁷ CB1 - sea water 4.5 x 10 ⁶ CB2 - sea water + DMSO Control for QUE-2 150 ppm CB2 - sea water + DMSO Control for β-Lapachona 150 ppm 1.4 x 10 ^b 1.1 x 10 ⁶ CB2 - sea water + DMSO Control for QUE-2 300 ppm CB2 - sea water + DMSO Control for β-Lapachona 300 ppm 4.5 x 10 ⁶ 1.4 x 10 ⁶ Antimicrobial Compositions Concentration (NMP / mL) (Biocidal Efficiency%) QUE-2 150 ppm <0.3 (100%) QUE-2 300 ppm <0.3 (100%) β-Lapachona 150 ppm 9.5 x 10 (99.99%) β-Lapachona 300 ppm 0.9 x 10 (> 99.99%) THPS 100 ppm certified <0.3 (100%)

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EXAMPLE 3 [0207] Evaluation of antimicrobial activity against cultures of

Planktonic BRS pool header P-19, and Desulfovibrio alaskensis, compared to compositions containing β-Lapachona, in concentration of 100 ppm, and Floretina, in concentration of 50 ppm, both having DMSO as vehicle, in contact time of 18 hours in microcosm consisting of deaerated synthetic seawater, proceeding to certification against the traditional commercial biocide GA at a concentration of 100 ppm.

[0208] The results presented in Table 8 and Figure 5 demonstrate relevance in the case of the composition containing Floretine, which presents Biocidal Efficiency, in a concentration of 50 ppm, in defiance of the two cultures tested BRS pool header P-19 and Desulfovibrio alaskensis, respectively, of 99.98% and 99.99% in the contact time of 18 hours.

[0209] Regarding the composition containing β-Lapachona, in a concentration of 100 ppm, relevant for this same period of contact, it was also observed to the extent that 100% Biocidal Efficiency was achieved for both cultures, in equivalence to assay certification against the traditional commercial biocide GA, in identical concentration.

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TABLE 8: Results of tests to evaluate the Biocidal Efficiency of compositions containing Floretine, β-Lapachona, in concentrations of 50 ppm and 100 ppm, respectively, in DMSO.

Planktonic BRS BRS Culture poolheaderP19 BRS Culture Disulfovibrium alaskensis Biological Controls BRS concentration (NMP / mL) Control of diluted 10 ' ¹ inoculated culture 1.4 x 10 ⁷ 4.5 x 10 ⁵ CB1 - sea water 1.4 x 10 ⁶ 1.4 x 10 ⁵ CB2 - sea water + DMSO Control for Floretina 50 ppm 4.5 x 10 ⁵ 1.4 x 10 ⁵ CB2 - sea water + DMSO Control for β-Lapachona 100 ppm 1.4 x 10 ⁶ 1.4 x 10 ⁵ Antimicrobial Compositions BRS concentration (NMP / mL) (Biocidal Efficiency%) Floretina 50 ppm 9.5 x 10 ¹ (99.98%) 0.7 x 10 ¹ (99.99%) β-Lapachona 100 ppm <0.3 (100%) <0.3 (100%) GA 100 ppm certification <0.3 (100%) <0.3 (100%)

EXAMPLE 4 [0210] Evaluation of antimicrobial activity in the presence of planktonic BRS pool cultures originating from PPS 32 and P-43 FPSO (Floating Production, Storage and Offloading) oil platform and BRS culture pool header P-19, compared to the compositions containing, respectively, QUE-2, β-Lapachona, and Floretina, having DMSO as a vehicle, each in a concentration of 300 ppm, in a contact time of 18 hours in a microcosm constituted by de-aerated synthetic sea water (Figure 6).

54/113 [0211] The results, presented in Table 9, demonstrate the biocidal action range, as well as inhibition in the H ₂ S producing metabolism attested by lead acetate indicator strips, in defiance of the BRS culture pools originating from the respective sea water injection systems, and treatment and separation of hydrocarbons and oily ballast from FPSO units.

TABLE 9: Results of tests to evaluate the Biocidal Efficiency of and inhibition in the H2S-producing metabolism of compositions containing QUE-2, βLapachona, Floretina in a concentration of 300 ppm, in DMSO.

Planktonic BRS Target culture BRS pool header P19 Target culture BRS pool P-32 Target culture BRS pool P-43 Biological Controls BRS concentration (NMP / mL) Control of diluted 10 ' ¹ inoculated culture 1.4 x 10 ⁷ - - Culture control without inoculated dilution - 4.5 x 10 ⁶ 4.5 x 10 ⁵ CB1 - sea water 1.4 x 10 ⁶ 1.4x 10 ⁵ 1.1 x 10 ⁵ CB2 - sea water + DMSO 300 ppm * 4.5 x 10 ⁵ 1.1 x 10 ⁵ 4.5 x 10 ⁴ Antimicrobial Compositions BRS concentration (NMP / mL) (Biocidal Efficiency%) QUE-2 300 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) β-Lapachona 300 ppm 9.5 x 10 ¹ (99.98%) <0.3 (100%) <0.3 (100%) Floretina 300 ppm <0.3 (100%) 2.5 x 10 ² (99.77%) <0.3 (100%) THPS 100 ppm certified <0.3 (100%) <0.3 (100%) <0.3 (100%)

* CB2 common to the three Antimicrobial Compositions tested [0212] The high specificity of antimicrobial action is confirmed and evidenced in relation to the BRS presented by the composition containing QUE-2, in contact time of 18 hours, and concentration of 300 ppm, demonstrating Biocidal efficiency of 100% (indicative of total death reach as assessed by the NMP technique) in relation to the three cultures tested, with emphasis on the susceptibility presented by the two cultures

55/113 originating from FPSO units, a scenario considered critical in the oil industry due to constant contamination by H ₂ S. The composition containing βLapachona, in a concentration of 300 ppm, also stands out for achieving a 100% Biocidal Efficiency in challenge to both cultures originating from FPSO units and the composition containing Floretine, in the same concentration of 300 ppm, presented Biocidal Efficiency of 100% in challenge to the target culture BRS pool P-43, of the respective FPSO unit and also in challenge to the BRS pool culture header P-19. It is evident, therefore, that the three BRS cultures tested were susceptible to the three compositions of the invention, with assay certification by the traditional commercial biocide GA at a concentration of 100 ppm, presenting 100% Biocidal Efficiency against the three target cultures in contact time 18 hours.

EXAMPLE 5 [0213] Evaluation of the antimicrobial activity of the composition containing

QUE-2 in the presence of the DMSO vehicle, after its dissolution in microcosm of deaerated synthetic seawater at a concentration of 300 ppm, in an environment also deaerated by NLO, and subsequent filtration in Millipore 0, 22 μιτι membrane, ensuring its sterilization, in challenge to the target culture BRS pool header P-19 in 18 hours (Figure 7).

[0214] Under the conditions traditionally applied to antimicrobial tests, the referred composition was tested, the results being shown in Table 10. These attest to the high Biocidal Efficiency of 100% (total death range by the NMP technique) compared to the composition containing QUE -2, even after filtration, indicating that the active substance is present in soluble form, and well preserved, in the contact microcosm constituted by de-aerated sea water, making sure the test against the traditional commercial biocide THPS.

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TABLE 10: Test results for assessing the Biocidal Efficiency of QUE-2 composition at a concentration of 300 ppm in DMSO.

Planktonic BRS BRS pool target culture header P-19 Biological Controls BRS concentration (NMP / mL) Control culture without dilution 1.1 x 10 ^b CB1 - sea water 1.1 x 10 ⁵ CB2 - sea water + DMSO Control for QUE-2 300 ppm (0.22pm membrane filtration) 4.5 x 10 ⁴ Antimicrobial Composition BRS concentration (NMP / mL) (Biocidal efficiency%) QUE-2 300 ppm (membrane filtration 0.22pm) <0.3 (100%) THPS 100 ppm certified <0.3 (100%)

EXAMPLE 6 [0215] The purpose of the test is to assess whether the composition containing QUE-2 in the presence of the DMSO vehicle meets the needs of the oil industry for effective biocidal action, at a concentration of 300 ppm, when in pre-solubilization conditions , simulating the marine environment itself, in shorter response times, such as 4 hours and, comparatively, in 24 hours, challenging the BRS pool culture typical of the FPSO unit environment.

[0216] In this experiment, the composition containing QUE-2 in DMSO at a concentration of 300 ppm, after dissolving in microcosm of synthetic sea water deaerated by NLO, in an environment also deaerated by NLO, is subjected to filtration in Millipore 0 membrane, 22 pm, certifying its sterilization, and subsequent evaluation of antimicrobial activity in defiance of the target cultures BRS pool header P-19 and BRS pool FPSO P-32, in planktonic form, in contact times of 4 hours and 24 hours , making sure against the traditional commercial biocide THPS at a concentration of 100 ppm.

[0217] The results observed in Table 11 demonstrate the achievement of high degrees of Biocidal Efficiency, indicating values of 100% or death

57/113 total (no detection of viable bacteria by the NMP technique) compared to the composition containing QUE-2 (Figure 8) filtered after its pre-dissolution in sea water at a concentration of 300 ppm. It is worth mentioning that the demonstration of susceptibility to that composition was obtained, under the conditions and contact times tested in relation to the two cultures of planktonic BRS. In addition to the expected operational advantage regarding the availability of the active substance soluble in sea water, these results are of great relevance for the oil industry, as the response time has been shortened to 4 hours, with identical effectiveness in the biocidal action.

TABLE 11: Results of tests to evaluate the Biocidal Efficiency of a composition containing QUE-2, at a concentration of 300 ppm in DMSO under pre-solubilization conditions in synthetic seawater.

Planktonic BRS Target culture BRS pool header P-19 Target culture BRS pool FPSO P-32 Biological Controls BRS concentration (NMP / mL) Control culture without inoculated dilution NR 1.4 x 10 ⁷ Culture-control dilution 10 ' ¹ inoculated 1.4 x 10 ⁶ NR Contact (hours) 4 h 24 h 4 h 24 h CB1 - sea water 2.5 x 10 ⁴ 1.1 x10 ^b 1.4 x 10 ^b 1.4 x 10 ^b CB2 - sea water + DMSO Control for QUE-2 300 ppm (membrane filtration 0.22pm) 4.5 x 10 ⁴ 4.5 x 10 ⁴ 4.5 x 10 ⁵ 4.5 x 10 ⁵ Antimicrobial Composition Concentration (Bioc Efficiency BRS (NMP / mL) : one way%) Contact (hours) 4 h 24 h 4 h 24 h QUE-2 300 ppm (0.22pm membrane filtration) <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%) THPS 100 certification PPm <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%)

NR - not done

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EXAMPLE 7 [0218] Evaluation of the antimicrobial activity of the composition containing

QUE-2, in a concentration of 300 ppm, in a microcosm consisting of synthetic seawater, against the targets BRS pool header P-19 and BANHT pool header P-19, in order to assess whether the said composition meets the need of the biocidal oil industry in shorter contact times.

[0219] The results observed in Table 12 demonstrate the achievement of the same degree of Biocidal Efficiency in the contact times of 4 hours and 24 hours, both against BRS and against BANHT (Figure 9), making sure the test against the biocide traditional commercial THPS (100 ppm). The target culture BRS pool header P-19 had also been shown to be susceptible to this composition, repeatedly, as in examples 1 to 3, when the contact period of 18 hours in the same microcosm was evaluated. It is also worth mentioning that the composition containing QUE-2 reached sterilizing potential (population reduction of 10 ⁶ with total death) for the BRS pool header P-19 culture in the contact times of 4 hours and 24 hours and, for the BANHT pool header culture P-19, said potential was reached in 24-hour contact. The relevance, therefore, of the results achieved, points to the potential application of the Composition containing QUE-2, in short response times (4 hours) to control consortium-forming planktonic organisms (BRS and BANHT), which as a rule enhance the generation of biofilms, also contributing to reduce the performance of biocidal substances.

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TABLE 12: Test result to evaluate the Biocidal Efficiency of Composition QUE-2 at a concentration of 300 ppm in DMSO.

BRS and BANHT Planktonic BRS pool target culture header P-19 BANHT target culture pool header P-19 Biological Controls Concentration (NMP / mL) Concentration (NMP / mL) Control cultures without dilution 1.5 x 10 ⁷ 1.1 x 10 ⁷ Contact (hours) 4 hours 24 hours 4 hours 24 hours CB1 - sea water 4.5 x 10 ⁵ 4.5 x 10 ⁵ 1.4χ10 ^δ 1.1 x 10 ^b CB2 - sea water + DMSO Control for QUE-2 300 ppm 1.1 x 10 ⁶ 1.4 x 10 ⁶ 4.5 x 10 ⁵ 1.1 x 10 ⁶ Composition QUE-2 QUE-2 300 ppm <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%) THPS 100 certification PPm <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%)

EXAMPLE 8 [0220] For the purpose of evaluating the performance of the compositions containing QUE-2, β-Lapachona, and Floretina, each with DMSO as a vehicle, in order to meet the needs of the oil industry with an efficient and fast biocidal action, that is, in lower concentrations combined with shorter contact times, an assay was developed to assess the antimicrobial activity of the referred compositions QUE-2, β-Lapachona, in concentrations of 100 ppm, 150 ppm and 300 ppm, and Floretine in concentrations 150 ppm and 300 ppm, and contact times of 4 hours and 24 hours in a microcosm consisting of synthetic seawater de-aerated by stripping by NLO, in defiance of the target culture BRS pool header P-19 in planktonic form. The certification was carried out against the traditional commercial biocide THPS, at a concentration of 100 ppm.

60/113 [0221] The results observed in Table 13 demonstrate the achievement of high degrees of Biocidal Efficiency, all with values higher than 99.9%, still achieving total death (100% in Biocidal Efficiency) in some of the experiments ahead to the referred compositions tested in the contact times of 4 hours and 24 hours, in the concentration range evaluated in defiance of the target culture of BRS pool header P-19 in planktonic form (Figure 10). This culture had already been shown to be susceptible to compositions containing QUE-2, Floretina and β-Lapachona, in the same microcosm, as in previous tests, when the contact period of 18 hours was evaluated, in concentrations of 150 ppm and 300 ppm. The relevance, therefore, of the results obtained, expressed in Biocidal Efficiency, points to the potential application of the compositions containing QUE-2 and β-Lapachona, in lower concentrations, reaching 100 ppm, as well as the composition containing Floretina in the concentration of 150 ppm in DMSO, obtaining efficacy also in short response times (4 hours) for controlling planktonic BRS.

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TABLE 13: Results of tests to evaluate the Biocidal Efficiency of compositions containing QUE-2, β-Lapachona, in concentrations of 100, 150 and 300 ppm in DMSO, and Floretina, in concentrations of 100 and 150 ppm in DMSO.

Planktonic BRS Target culture BRS pool header P-19 Biological Controls BRS concentration (NMP / mL) Control culture without inoculated dilution 4, 5x 10 ⁷ Contact (hours) 4 hours 24 hours CB1 - sea water 4.5 x 10 ⁶ 4.5x10 ⁶ CB2 - sea water + DMSO Control for QUE-2 and β-Lapachona 100 ppm 4.5 x 10 ⁶ 4.5 x 10 ⁶ CB2 - sea water + DMSO Control for QUE-2 and β-Lapachona 1.1 x 10 ⁷ 4.5 x 10 ⁶ 150 ppm CB2 - sea water + DMSO Control for Floretina 150 ppm 1.1 x 10 ⁶ 1.4 x 10 ⁶ CB2 - sea water + DMSO Control for QUE-2 and β-Lapachona 1.1 x 10 ⁷ 1.1 x 10 ⁷ 300 ppm CB2 - sea water + DMSO Floretine Control 300 ppm 1.1 x 10 ⁶ 1.4 x 10 ⁶ Antimicrobial Compositions BRS concentration (NMP / mL) (Biocidal Efficiency%) Contact (hours) 4 hours 24 hours QUE-2 100 ppm 1.4 x 10 ³ (99.97%) 7.5 x 10 (> 99.99%) QUE-2 150 ppm 2.5 x 10 ² (> 99.99%) <0.3 (100%) QUE-2 300 ppm <0.3 (100%) <0.3 (100%) β-Lapachona 100 ppm 9.5 x 10 (> 99.99%) 6.0 x 10 (> 99.99%) β-Lapachona 150 ppm <0 3 <0 3 (100%) (100%) β-Lapachona 300 ppm <0.3 (100%) <0.3 (100%) Floretina 150 ppm 2.5 x 10 ¹ (> 99.99%) <0.3 (100%) Floretina 300 ppm 0.6 x 10 ¹ (> 99.99%) <0.3 (100%) THPS certified (100 ppm) <0.3 (100%) <0.3 (100%)

62/113 [0222] This test consolidates the results previously presented, in terms of Biocidal Efficiency (%) in challenge to the target culture of BRS pool header P-19, in planktonic form, demonstrating its susceptibility to the compositions of present invention comprising QUE-2, β-Lapachona, and Floretina, which were tested in conservative profiles, both in concentrations and response times to biocidal action, with remarkable results in the control of planktonic BRS originating from water injection system of the oil industry. It should also be noted that, according to strict criteria in the literature for the medical-pharmaceutical area (sterility assurance I took (SAL) of 1 (T ⁶ ) [von WOEDTKE et al. “The limits of sterility assurance”. GMS Krankenhhyg Interdiszip. 3 ( 3): Doc19. 2008], the Sterilizing Character (CE) was achieved by certifying death in the presence of compositions, under determined conditions (contact time and concentration) through non-detection of the target culture BRS pool header P19 according to the population decrease of 10 ⁶ , using the NMP technique, when compared to the respective Biological Control (CB2) of the culture, Table 14 illustrates this important characteristic common to compositions containing QUE-2, β-Lapachona, and Floretina, in DMSO, in challenge to the BRS pool header P19 culture in planktonic form.

TABLE 14: Sterilizing character of the compositions comprising QUE-2, βLapachona, Floretina, in concentrations of 150 ppm and 300 ppm in DMSO.

STERILIZING CHARACTER Composition Antimicrobial Concentration 150 ppm Concentration 300 ppm Contact (hours) 4 h 24 h 4 h 24 h QUE-2 AT CE CE CE β-Lapachona CE CE CE CE Floretina AT CE AT CE

NA - Sterilizing character not achieved according to the defined criteria

CE- Sterilizing Character achieved according to the defined criteria [0223] Great relevance was also observed as the results compared to the compositions of the invention, especially when they present Sterilizing Character (CE), in the test conditions explained

63/113 above, which would be comparable to the traditional commercial biocide THPS. Therefore, the feasibility of applying the compositions of the present invention to injection and production systems is pointed out, as well as to other inshore and offshore oil installations, aiming at the prevention, mitigation and / or control of biogenic processes mediated by metabolic action and / or uncontrolled growth of BRS, such as CIM, souring and plugging wells. Table 15 presents a synthesis of results related to Biocidal Efficiency (%) achieved in defiance of the target culture of BRS pool header P-19 in planktonic form compared to compositions containing QUE-2 and β-Lapachona in DMSO, according to profiles concentration (100 ppm, 150 ppm and 300 ppm) and Floretine (150 and 300 ppm) in DMSO in varying contact time (4 h, 18 h and 24 h) in de-aerated sea water microcosm, compared to the biocide traditional commercial THPS (100 ppm).

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TABLE 15: Results of tests to evaluate the Biocidal Efficiency of compositions containing QUE-2, β-Lapachona, in concentrations of 100, 150 and 300 ppm in DMSO, and Floretina, in concentrations of 100 and 150 ppm in DMSO.

BIOCIDIC EFFICIENCY (%) Contact 4 hours Contact 18 hours Contact 24 hours Composition containing QUE-2 QUE-2 (100 ppm) 99.97% NR > 99.99% QUE-2 (150 ppm) > 99.99% 100% 100% QUE-2 (300 ppm) 100% 100% 100% Composition β-Lapachona β-Lapachona (100 PPm) > 99.99% NR > 99.99% β-Lapachona (150 Ppm) 100% 99.99% 100% β-Lapachona (300 PPm) 100% > 99.99% 100% Composition Floretina Floretine (150 ppm) > 99.99% 100% 100% Floretine (300 ppm) > 99.99% 100% 100% Trac commercial biocide icional - Certification THPS (100 ppm) 100% 100% 100%

NR - Not done

EXAMPLE 9 [0224] Evaluation of the Biocidal efficiency of compositions of the present invention and their influence on CIM in defiance of the BRS target culture in the process of adhesion to SAE 1020 carbon steel biocoupons arranged in a microcosm composed of de-aerated seawater by stripping with NLO, through the Sensor Assay Methodology (MES - newly formed biofilm).

65/113 [0225] This Methodology (ANDRADE, AC et al. “Conditioning seawater for hydrostatic testing in pipelines. 15 ^to MR-MIC Cenpes-Petrobras. 1993), simulates, under static conditions, a typical marine anoxic environment of the oil industry, when a biocidal substance is added to the respective fluid (deaerated synthetic seawater, for example), in the presence of BRS originally planktonic in a possible process of adhesion to the metallic surface (biocoupon). This examines the potential for inhibiting BRS adhesion, in sessile form, compared to the compositions of the invention.

[0226] This protocol allows comparing, in a comparative way, compositions in terms of the reach of Biocidal Efficiency in relation to planktonic BRS, and sessile, in the period of contact of fifteen (15) days, in conjunction with the ultrastructural evaluation by Microscopy technique Scanning Electronics (SEM) of the newly formed biofilm on the surface of biocoupons for 18 hours of contact. As well as allowing to determine: parameters of uniform corrosion (loss of mass), which are evaluated in periods of contact fifteen (15) days and forty-five (45) days, and parameters of localized corrosion (presence and depth of pits) , which are observed, and evaluated, if detected, in both periods.

[0227] The laboratory procedure (handling of cultures, biocoupons, compositions of the present invention and traditional commercial biocides, solutions and additional inputs) is performed under sterile conditions in a vertical laminar flow chamber, under continuous purge of oxygen-free nitrogen (NLO ). All experiments are performed in triplicate and in relation to their respective Biological Controls (deaerated synthetic seawater and deaerated synthetic seawater added, in defined volumetric equivalents of the DMSO vehicle), which is applied to the compositions of the invention, and also, comparatively, in relation to positive controls compared to traditional commercial biocides THPS and GA. Sterile controls (Chemical Controls / QC) are designed for each experiment.

[0228] Briefly, the MES (newly formed biofilm) assay is performed by immersing the pre-sterilized carbon steel specimen (biocoupon) by electron beam technique (E-beam) in the microcosm of

66/113 contact constituted by pre-sterilized sea water deaerated by NLO and in the presence of resazurin, visual indicator of oxy-reduction, with a view to the observation of anaerobiosis. In sequence, in separate sets of experiments, each of the compositions of the invention is added to the concentration (1000 ppm) and, comparatively, each of the traditional commercial biocides (500 ppm), followed by immediate inoculation, in independent experiments, of the culture of BRS pool header of P-19 and of the control culture Desulfovibrio alaskensis, both in planktonic form, and with a culture age of 18 hours after growth in modified Postgate C medium, and in a concentration equal to or greater than 10 ⁷ organisms / mL, according to the NMP technique. This is followed by the Contact step carried out by incubation at a temperature of 30 ° C in anaerobic conditions according to the determined periods, of 15 days and 45 days, in order to also allow the evaluation of the corrosive process. In sequence, to determine Biocidal Efficiency with respect to sessile and planktonic BRS, respectively, proceed to the Microbial Growth Revelation stage, by incubation at 30 ° C in modified Postgate E medium, of biocoupons (newly formed biofilms) ), and the liquid phase of the crops, according to the NMP technique, recommending observation of growth for a period of 28 (twenty-eight) days (from Postgate, 1984). The number of replicates is defined in order to integrate, and to carry out, all experiments, and their respective Chemical (absence of culture) and Biological (absence of substances with biocidal characteristics) controls, in conjunction with contact times (15 and 45 days) compared to the compositions of the invention.

[0229] Table 16 summarizes information related to the identification applied to Biological (CB) and Chemical (CQ) Controls, to the characteristics and specifications of the metallic specimen (biocoupon), to the experiments matrix and other applied codes.

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TABLE 16: Information on the identification applied to biological and chemical controls, biocoupon, experimental matrix and Characteristics of the metallic specimen (biocoupon).

Biological and Chemical Controls

CB1 - Sea water + BRS culture

CB2 - Sea water + BRS culture + DMSO (vehicle volumetric equivalent)

CQ1 - Sea water

CQ2 - Sea water + DMSO (vehicle volumetric equivalent)

CQ3 - Seawater + Composition Biocide or traditional commercial biocide

Characteristics of the metallic specimen (biocoupon) SAE 1020 steel specification - Elemental chemical composition (% w / w) Race number Carbon Manganese Silicon Phosphor Sulfur 89406481 0.190 0.520 0.160 0.012 0.012 Dimensions: (39.0 x 11.0 x 3.0) mm Treatment: jet of glass microspheres (0 = 600/320 pm), with pre-pickling and weighing according to ASTM G1 - 03 (2011) - Standard Practice for Preparing, Cleaning, and Evaluating Corrosion Test Specimens Sterilization: The sterilization of biocoupons is performed using the electron beam exposure technique (E-beam) using two linear accelerators with 18 kW of power and 10 MeV of energy in each one with an applied dose of 5.0 kGy, the which was previously defined in laboratory experiments focusing on the effective dose of death in relation to the said target cultures. The certification was carried out by determining the effective dose of sterilization at a value equal to or greater than 2.5 kGy, which was defined by the reach of total death (detection level below 0.3 organisms / mL, according to the NMP technique) of said BRS target cultures in sessile form. The technique (E-beam) is approved by the FDA / USA for use in medical equipment, including, and its principle is based on scanning the flow of electrons through the substrate to be sterilized, with the death of the contaminating population possibly present by mechanism of direct breaking of microbial DNA chains, with the generation of secondary particles, such as free radicals, which are highly reactive compounds that react with microorganisms, preventing their reproduction.

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EXAMPLE OF EXPERIMENT MATRIX 0 MES - newly formed biofilm (15 and 45 days) T = 30 C, sets of triplicates Antimicrobial compositions of the invention or traditional commercial biocides x BRS target cultures Test identification * MES - identification of Compositions Antimicrobials of the invention in DMSO Concentration in sea water (PPm) Culturactrol BRS Desulfovibrio alaskensis BRS pool target culture header P-19 ALQ HQ -> Q 1000 ALB HB -> B 1000 ALPH HPH XPH 1000 Culture control Desulfovibrio alaskensis BRS pool target culture header P-19 MES - identification of biocides commercials Concentration (PPm) SEAWEED HGA Glutaraldehyde -> GA 500 ALT HT THPS AT 500 Culture control Desulfovibrio alaskensis BRS pool target culture header P-19 MES - identification of Controls Biological DMSO added - ALCB! HCBi Sea water desaerada -> CBi ALCB ₂ hcb ₂ De-aerated seawater + DMSO equiv. -> CB ₂ Equivalent volume MES - identification of Chemical Controls (sterile) CQ1 Sea water shameless CQ2 Sea water deaerated + DMSO equiv. Equivalent volume CQ3Q Sea water desaerada + Q Equivalent volume CQ3B Sea water shabby + B Equivalent volume CQ3PH Sea water desaerada + PH Equivalent volume CQ3 GA Biocide GA Equivalent volume CQ3T Biocide THPS Equivalent volume

* Identification symbols:

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AL- control culture BRS Desulfovibrio alaskensis

H- target culture BRS pool header P-19

Q- Composition containing QUE-2 in DMSO B- Composition containing β-Lapachona in DMSO PH- Composition containing Floretine in DMSO CQ- Chemical Controls (sterile)

CB- Biological Controls

CB1- Microcosmic Biological Control in de-aerated seawater with NLO CB2- Microcosm Biological Control in de-aerated seawater with NLO + DMSO equivalent

Control of microbial growth of planktonic BRS according to newly formed MESbiofilm [0230] Table 17 (reduction of the population of planktonic BRS) shows significant growth control of the two cultures of planktonic BRS, in contact time of 15 days, under anoxic conditions of the fluid (synthetic sea water de-aerated with NLO), at a temperature of 30 ° C, and in the presence of metallic specimens (biocoupons) of SAE 1020 carbon steel. The reduction in the bacterial population, measured by the NMP technique, was in the range from 10 ⁴ to 10 ^{6 in} relation to the respective control population, when compared to the compositions of the present invention, showing the following classification in decreasing order of Biocidal Efficiency: QUE-2, βLapachona and Floretina. It should be noted that the Biocidal Efficiency of the composition containing QUE-2 was equivalent to those presented by the traditional commercial biocides GA and THPS for the target culture BRS pool header P-19 and, superior in challenge to the control culture BRS Desulfovibrio alaskensis, regarding their respective Biological Controls. The concentration of BRS initially present in the contact (NMP in the range of 10 ⁶ to 10 ⁷ NMP / mL) in defiance of the referred compositions and the population decrease achieved (total death) in relation to them are conservatively compatible with the requirements and norms of the NCCLS [ Methods for antimicrobial susceptibility testing of anaerobic bacteria; approved standard-sixth edition (M11-A6). Wayne: NCLLS. 2004)] applied to the selection of antibiotics for human use in the control of diseases caused by pathogenic anaerobic bacteria and, still, to the recommendations of the reference article

70/113 (COS, P. et al. “Anti-infective potential of natural products: how to develop a stronger in vitro proof-of-concept.” J. Ethnopharmacol. Vol. 19; 106 (3): 290-302 2006) about the applied inoculum concentration, confirming the conservative character of the assay. Also relevant is the fact that the population decrease of 10 ⁶ was achieved compared to the Composition containing QUE-2, when in defiance of the two planktonic cultures of BRS. The present result can be compared to the sterility assurance level (SAL) of 1CT ⁶ [von WOEDTKE et al. "The limits of sterility assurance". GMS Krankenhhyg Interdiszip. 3 (3): Doc19. 2008], indicating the reach of the Sterilizing Character (CE).

TABLE 17: Evaluation of the compositions of the invention and commercial biocides for reducing the population of planktonic BRS.

Antimicrobial Compositions invention and traditional commercial Biocides (contact time - 15 days) Reduction of planktonic BRS population (target culture pool header P-19) Reduction of planktonic BRS population (control culture Desulfovibrio alaskensis) Q 6 10 * 6 10 * BL 5 10 * 5 10 * PH 5 10 * 4 10 * GA 6 10 ** 5 10 ** T 6 10 ** 5 10 **

* Population Reduction (MPN / mL) in the presence of the Compositions

Antimicrobials of the invention in relation to CB2 Biological Control ** Population Reduction (NMP / mL) in the presence of traditional commercial biocides in relation to CB1 Biological Control

Evaluation of Biocidal Efficiency against the compositions of the invention and traditional commercial biocides in inhibiting sessile BRS (MES - newly formed biofilm)

71/113 [0231] It was observed, by visual analysis performed after the

Disclosure in the presence of the modified Postgate E medium, positive indication of BRS growth in sessile form, associated with the blackening of the biocoupon (characteristic biofilm formation in the presence of FeS), with delay in the detection time to 148 hours (six days) compared to the tested compositions of the invention, when compared to the respective Biological Controls, which showed positive detection in 48 hours, which would indicate less activity in the growth (inhibition) of sessile BRS compared to the referred compositions. According to the same visual analysis, the traditional commercial biocides GA and THPS showed insignificant partial growth after 15 days (a triplicate of a set) only in the case of the culture control BRS Desulfovibrio alaskensis, maintaining inhibition at 28 days. The results in the presence of traditional commercial biocides confirm previous studies carried out (laboratory and field) that indicate acceptable performance of these products in combating BRS in the process of adhesion to metallic surfaces, especially GA (ANDRADE, CA et al. “Additives used in water for hydrostatic test in pipelines - Part I "and ANDRADE, CA et al." Selection of additives used in sea water for hydrostatic test in pipelines - Phase II ". Cenpes- Petrobras. 11 ^to MR-MIC. 1989). Table 18 presents the referred results in the Disclosure stage.

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TABLE 18: Results of the evaluation of Biocidal Efficiency against compositions of the invention and traditional commercial biocides in inhibiting sessile BRS.

Revealing the growth of sessile BRS in the presence of the modified Postgate E medium (visual observation of the biocoupon after 15 days of contact with the Antimicrobial Compositions of the invention and traditional commercial biocides)

Controls Biological, Antimicrobial Compositions of the Invention and Biocides traditional commercials Target culture BRS pool header P-19 BRS control culture Alaskensis desulfovibrio Triplicates Triplicates THE B Ç THE B Ç CBi (+) 48 H (+) 48 h (+) 48 h (+) 48 h (+) 48 h (+) 48 h cb ₂ (+) 48 H (+) 48 h (+) 48 h (+) 48 h (+) 48 h (+) 48 h Q (+) 148 h (+) 148 H (+) 148 h (+) 148 h (+) 148 h (+) 148 H BL (+) 148 h (+) 148 h (+) 148 H (+) 148 h (+) 148 h (+) 148 H PH (+) 148 h (+) 148 h (+) 148 H (+) 148 h (+) 148 h (+) 148 H GA (-) 28 days (-) 28 days (-) 28 days (+) 15 days (-) 28 days (-) 28 days THPS (-) 28 days (-) 28 days (-) 28 days (-) 28 days (+) 15 days (-) 28 days

Visual observation of blackening of the biocoupon, attributing to the formation of FeS, indicating possible association with the adhesion of BRS originally inoculated to the MES experiment in planktonic form, becoming sessile

73/113 (+) positive growth after the reported period (-) negative growth until the reported period [0232] In order to confirm, by anaerobic microbiology methods previously applied to the tests challenging the planktonic BRS cultures, the results relative to the visual observation of the growth of sessile BRS, a step was taken to re-reveal the biocoupons originating from the experiments against the compositions of the invention and traditional commercial biocides in the Methodology of the Sensing Tests (MES), and their respective Chemical Controls (QC) and Biological (CB1 and CB2), after incubation in modified Postgate E medium. This protocol was thus designed in an experimental way, being applied in sequence to the Revelation stage and the respective visual observations, being presented, briefly, below.

[0233] The re-development stage was carried out in a laminar flow chamber under sterile conditions, with each biocoupon being washed with a reducing saline solution (SRR) and scraping the apparent biofilm under NLO flow, with preservation of the suspension. resulting in SSR pre-purged with NLO and subsequent inoculation of defined aliquots of said homogeneous suspension in Postgate E medium modified for evaluation according to the NMP technique of the growth of BRS supposedly adhered to the biocoupon, confirming, or not, the presence and concentration of BRS in sessile form at this stage of the MES experiment. Briefly, in this experimental stage, the analysis of planktonic bacteria was carried out directly in the liquid phase of the experiment pool, which consisted of an equal volume of 10 mL of two of three triplicates. The analysis of sessile bacteria was performed in the liquid phase consisting of the suspension of the biofilm in a reducing saline solution (SSR), which was obtained from scraping each respective biocoupon (total area 11, 58 cm ² ) under NLO volume flow 10 mL each. The evaluation, both of the growth of planktonic BRS (liquid phase) and of sessile (biofilm in adhesion to biocoupon), was carried out according to the NMP technique (from Postgate, 1984) after the contact period of 15 days in front of the respective compositions of the invention (concentration of 1000

74/113 ppm) and traditional commercial biocides (500 ppm), followed by the incubation period with growth observation in modified Postgate E medium in up to 30 days. It was considered that the detected viable BRS concentrations, when <0.3 NMP / mL, because they are below the detection limit of the NMP technique, would not be measurable and can therefore be considered, when compared to Biological Controls in bands recommended (10 ⁴ to 10 ⁷ NMP / mL), as tending to zero and thus expressing the Biocidal Efficiency achieved of total bacterial death. In Table 19 and Figures 11A (for BRS pool header P-19) and 11B (BRS Desulfovibrio alaskensis), the results are presented, observing the Biocidal Efficiency for each respective Biological Control. The results obtained from this experimental protocol demonstrate that the compositions of the invention are effective in terms of the control of planktonic and sessile bacteria (newly formed biofilm) in relation to the two cultures tested.

TABLE 19: Results of tests to evaluate the Biocidal Efficiency of compositions comprising QUE-2, β-Lapachona, Floretina, in a concentration of 1000 ppm in DMSO against planktonic and sessile bacteria.

(*) Target culture BRS pool header P-19 Control culture BRS Desulfovibrio alaskensis (**) Planktonic BRS a Sessile BRS (biocoupon) (suspension) Planktonic BRS a Sessile BRS (biocoupon) (suspension) Biological Controls BRS concentration (NMP / mL) CB1 1.5 x 10 ^ϋ 4.5 x 10 ⁴ 1.4 x 10 ^b 4.5 x 10 ⁴ CB2 1.4 x 10 ⁵ 1.4 x 10 4.5 x 10 ⁴ 1.1 x 10 ^b Antimicrobial Compositions of the Invention BRS concentration (NMP / mL) (Biocidal Efficiency%) QUE-2 1000 ppm <0.3 (100%) <0.3 (100%) 0.4 (> 99.99%) <0.3 (100%) β- Lapachona 1000 ppm 9.5 x 10 ¹ (99.93%) 0.4 (> 99.99%) 2.5 (99.99%) 1.4 x 10 ³ (98.73%) Floretine 1000 ppm <0.3 (100%) 9.5 x 10 ¹ (99.93%) NR 2.5 x 10 ³ (97.73%) GA 500 ppm certification <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%) THPS 500 ppm certified <0.3 (100%) <0.3 (100%) <0.3 (100%) <0.3 (100%)

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NR- not carried out due to the non-availability of samples of the referred culture (*) MES experiments (15-day contact against Antimicrobial Compositions and traditional commercial biocides) (**) Experimental re-development test carried out after incubation period 30 days (Development in modified Postgate E medium), when growth is assessed visually) [0234] The compositions containing QUE-2 and βLapachona were therefore classified for the control of the two cultures of BRS tested, in planktonic form, and for the control of culture pool header P-19, also the composition containing Floretine, according to criteria of Biocidal Efficiency equal to or greater than 99.9%. To control the sessile form, for the two cultures tested, the Biocidal Efficiency of the composition containing QUE-2 is confirmed and, in the case of the target culture pool header P-19, the Biocidal Efficiency of the compositions containing β- is also confirmed. Lapachona and Floretina. The traditional commercial biocides GA and THPS demonstrated Biocidal Efficiency of 100% for the two cultures tested, in planktonic and sessile forms. Thus, it is considered that the proposed experimental protocol (re-disclosure), applied to MES (newly formed biofilm), proved to be certified in relation to the two BRS cultures challenged against the compositions of the present invention (Figures 11A and 11B ).

MES (newly formed biofilm) - Evaluation of parameters associated with the corrosive process influenced by microorganisms (CIM)

Sulfide control in the liquid phase [0235] An experimental technique of Sulfide Evolution by acid attack was applied, as described in the patent specification PI 0904216-4, which, briefly, includes the absorption of the solubilized sulfide by acid action (HCI 6N, hot), followed by NLO drag, precipitation in alkaline cadmium sulfate solution, and spectrophotometric quantification of the resulting sulfide by the Methylene Blue method. This protocol was carried out in the liquid phase (pool of triplicates) of contacts with the compositions of the invention and traditional commercial biocides, immediately after the removal of the respective biocoupons for analysis.

76/113 concerning the parameters of the corrosive process. The estimate of the percentage of inhibition in the generation of sulfide, a typical BRS metabolite, was carried out by comparing the tests in the presence of the compositions of the invention and traditional commercial biocides to their respective Biological and Chemical Controls. Table 20 shows, considering the contact period of 15 days, a trend of greater reach in the inhibition of sulfide of the liquid phase compared to the compositions of the present invention by the control culture BRS Desulfovibrio alaskensis, with little inhibition, or null inhibition. in the case of traditional commercial biocides, as was also the case for the BRS pool header P-19 culture more clearly. It is worth mentioning that, in this period, the growth control of planktonic bacteria was also observed in relation to the compositions of the present invention and to traditional commercial biocides (results previously presented and discussed). It is believed that, in this initial phase, the process of adhesion of BRS originally planktonic to the biocoupon may have started when in the presence of compositions of the present invention and / or of the traditional biocides GA and THPS, more attenuated when compared to the Biological Controls related. In the contact period of 45 days, however, there is a tendency towards stabilization regarding the inhibition of sulfide in the liquid phase, for the traditional commercial biocides GA and THPS and for the compositions of the present invention, except for the composition containing Floretine, in this case. only in relation to the control culture BRS Desulfovibrio alaskensis, which showed less inhibition (26.92%), and the biocide THPS (57.89%) in this case for the target culture BRS pool header P-19. It is worth noting the high reach in the inhibition of the liquid phase sulfide in the presence of the composition containing QUE-2 for both tested cultures, Desulfovibrio alaskensis (76.19%) and pool header P19 (78.57%), as well as in the presence of composition containing β-Lapachona for the culture Desulfovibrio alaskensis (78.94%). It should also be noted that, during this 45-day period, corrosion reactions should stabilize in an anoxic environment, which does not tend to occur in a shorter contact period of 15 days. In Table 20, the referred results are presented.

TABLE 20: Results of tests to evaluate parameters associated with the corrosive process influenced by microorganisms (MIC), based on

77/113 percentage of inhibition in the generation of sulfide.

Inhibition of sulfide generation in the presence of the Antimicrobial Compositions of invention and traditional commercial biocides (%) Experiment identification 15 days 45 days HQ 42.42 78.57 HBL 18.68 69.23 HPH 70.9 69.70 HGA 3.4 76.19 HT ~ zero 57.89 ALQ 63.26 76.19 ALBL 53.22 78.94 ALPH 70.05 26.92 SEAWEED 16.05 74.13 ALT ~ zero 76.78

Note: evaluation carried out considering the detection of evolved H + sulfide (triplicate pool - mass base) of each Sensor Assay in the presence of the Antimicrobial Compositions of the present invention and the traditional commercial biocides in relation to the sum of the respective Chemical (QC) and Biological Control (CB) (triplicate pool - mass base) in each contact period.

Evaluation of Corrosion Influenced by Microorganisms (CIM) [0236] The corrosion parameters evaluated from the protocol of the

MES (newly formed biofilm) compared to the compositions of the present invention and, compared to traditional commercial biocides, in the presence of BRS (biotic assays) and in absence (abiotic assays), which will be presented and discussed below, were obtained from of ASTM Standards (ASTM G1 - 03 (2011) - “Standard Practice for Preparing, Cleaning, and Evaluating Corrosion Test Specimens and ASTM G31-72 (2004)“ Standard Practice for Laboratory Immersion Corrosion Testing of Metals, with necessary adaptations to the characteristics of the assay in the presence of bacteria.

Mass loss test [0237] Briefly, the specimens (biocoupons), with determined surface area of contact with the liquid media (microcosm), were

78/113 cleaned with acetone for 15 minutes, on ultrasound, and weighed on a calibrated digital analytical balance, accurate to 0.01 mg, to record its initial mass. The packaging was done in a desiccator with NLO atmosphere, to preserve its surface, followed by sterilization by electron beam technique (E-beam). These biocoupons were subjected to tests previously described in the Methodology of the Sensing Assays (MES - newly formed biofilm), having been incubated at a temperature of 30 ° C, under anoxic conditions (purging with NLO), against the compositions of the present invention and the biocides traditional commercials, in microcosm consisting of synthetic seawater deaerated by purging with NLO (biotic and abiotic experiments, and their respective Biological and Chemical Controls) during contact periods of 15 and 45 days, in static regime, after which they were evaluated according to ASTM standard. Three (3) metallic specimens (biocoupons) were tested for each condition / experiment.

[0238] The results of the uniform corrosion rate (mm / year) evaluated by weight loss, related to the Sensing Tests in relation to the compositions of the invention and the traditional commercial biocides, and to their respective Chemical and Biological Controls, expressed in their average values , weighting the standard deviations, can be seen in Table 21 for the two periods of contact with the said biocidal solutions, 15 and 45 days, and for the two BRS cultures tested.

[0239] According to the classificatory character of degrees of risk, which is applied to the assessment of the aggressiveness of injection waters, in terms of generalized corrosion and localized corrosion (pits) as established in Table 22, graphs were constructed (Figures 12 A, BC and D), demonstrating the trend presented by the results regarding these criteria.

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TABLE 21: Results of tests to assess the corrosive process influenced by microorganisms (MIC) based on mass loss.

MES - newly formed biofilm Experiment identification Average uniform corrosion rate ± standard deviation (mm / year) 15 days 45 days Controls Biological HCB ₁ 0.02473 ± 0.00456 0.00728 ± 0.00201 AICB ₁ 0.02816 ± 0.00493 0.00747 ± 0.00243 hcb ₂ 0.02349 ± 0.00198 0.01018 ± 0.00333 aicb ₂ 0.02634 ± 0.00590 0.00965 ± 0.00091 Controls Chemicals QC ₁ 0.05727 ± 0.00744 0.02092 ± 0.01858 cq ₂ 0.05089 ± 0.01726 0.01051 ± 0.00218 cq ₃ q 0.02192 ± 0.00479 0.02192 ± 0.00255 cq ₃ bl 0.04172 ± 0.01408 0.01298 ± 0.00592 CQ ₃ PH 0.02803 ± 0.00444 0.02875 ± 0.00344 CQ ₃ GA 0.06910 ± 0.01807 0.05337 ± 0.00852 cq ₃ t 0.04519 ± 0.01470 0.03972 ± 0.01240 Antimicrobial compositions of the present invention and traditional commercial biocides HQ 0.02517 ± 0.00139 0.00653 ± 0.00026 ALQ 0.02030 ± 0.00174 0.00581 ± 0.00067 HBL 0.03142 ± 0.00043 0.00996 ± 0.00000 ALBL 0.02978 ± 0.00761 0.01176 ± 0.00272 HPH 0.03293 ± 0.00311 0.01113 ± 0.00132 ALPH 0.02045 ± 0.00059 0.01527 ± 0.00235 HGA 0.01902 ± 0.00072 0.00758 ± 0.00265 SEAWEED 0.02029 ± 0.01125 0.00810 ± 0.00510 HT 0.02858 ± 0.00365 0.00739 ± 0.00140 ALT 0.02301 ± 0.00866 0.00935 ± 0.00298

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TABLE 22 :. Injection water aggressiveness criteria

Injection water aggressiveness criteria Uniform corrosion rate Corrosion rate by pits Fluid corrosivity - degrees of risk mm / year mm / year <0.025 <0.305 LOW 0.025-0.125 0.305-0.610 MODERATE 0.126-0.254 0.635 - 2.438 SEVERE > 0.254 > 2,438 VERY SEVERE

(Injection Water Specification. 2- Technical Meeting on Wastewater Treatment and Injection Water. Cenpes-Petrobras. 1997)

Localized corrosion [0240] The tendency to localized corrosion, with the observation of the presence of pits, was detected only in an exposure time of 45 days. Table 23 presents a demonstration of their occurrence and their respective depths in relation to the Sensing Assays against the compositions of the present invention and traditional commercial biocides in the presence of the two cultures tested and their respective Biological and Chemical Controls. An estimate of the “equivalent” localized corrosion rate was established, based on the exposure time (period of 45 days) in which the referred pits were observed, converting them to base-year according to the reported depth (μ), defining, in this way considered conservative, the possible degrees of risks in the light of the criteria previously presented to assess the aggressiveness of the injection water.

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TABLE 23: Test results for the evaluation of localized corrosion.

MES - newly formed biofilm Assay identification Localized corrosion (pitting) (contact 45 days) Observation of pit (s) deepest (s) (μ) “Equivalent” localized corrosion rate (mm / year) - degrees of risk Chemical Controls QC, 4 and 6 cps = 40 μ 0.3244 MODERATE CQ _Z cp 6 = 30 μ 0.2433 LOW CQ ₃ Q cp 4 = 30μ 0.2433 LOW cq ₃ BL 4 and 5 cps = 30 μ 0.2433 LOW cq ₃ ph OND OND OND cq ₃ ga OND OND OND cq ₃ t cp 4 = 20 μ 0.1622 LOW Biological Controls HCBt 4, 5 and 6 cps = 40μ 0.3244 MODERATE hcb ₂ OND OND OND ALCBi cp 4 = 50 μ 0.4056 MODERATE ALCB2 cp 4 and 5 = 40 μ 0.3244 MODERATE Antimicrobial compositions of the present invention and traditional commercial biocides HQ cp 5 = 40 μ 0.3244 MODERATE HBL cp 4 = 50 μ 0.4056 MODERATE HPH 4 and 5 cps = 45 μ 0.3650 MODERATE HGA 5 and 6 cps = 45μ 0.3650 MODERATE HT cp 4 = 50 μ 0.4056 MODERATE ALQ cp 6 = 40 μ 0.3244 MODERATE ALBL 4.5 and 6 cps = 40 μ 0.3244 MODERATE ALPH 4 and 5 cps = 45 μ 0.3650 MODERATE SEAWEED cps 4, 5 and 6 = 45 μ 0.3650 MODERATE ALT cp 6 = 65 μ 0.5272 MODERATE

cp- identification of the metallic specimen (biocoupon) in which the pit was observed, measuring its depth (μ)

OND - observation not available

Evaluation of the uniform corrosive process

82/113 [0241] The 45-day exposure period applied to both the determination of uniform corrosion (loss of mass - »mm / year) and the estimate of localized corrosion (depth of pits -» equivalence in mm / year risk of perforation), proved to be more suitable for assessing the aggressiveness of the compositions of the present invention examined in relation to their respective controls, as well as traditional commercial biocides. In this period of 45 days, Tables 24 and 25 with results of the uniform corrosion rate (mm / year) show that the referred compositions in the presence of the two evaluated cultures showed a tendency towards uniform corrosion characterized as low (values of corrosion rate <0.025 mm / year according to Table 22 of Injection water aggressiveness criteria), especially in the case of the composition containing QUE-2 (corrosion rates in mm / year of 0.00653 ± 0.00026 and 0.00581 ± 0.00067 relative , respectively, to the BRS pool header P-19 and Desulfovibrio alaskensis cultures), observed in relation to their respective Biological Controls and also when compared to the other compositions of the present invention and / or to traditional commercial biocides.

[0242] It should be noted that in the case of localized corrosion (occurrence of pitting) the composition of the present invention comprising QUE-2, observed in relation to their respective Biological Controls and compared to the other compositions of the present invention and / or traditional commercial biocides, presented a lower “equivalent” corrosion rate per pitting for both tested cultures (0.3244 mm / year), considering only the highest pitting detected, which gives a moderate degree of risk. Expectation of uniform corrosion protection [0243] The expectation of uniform corrosion protection according to the performance of the compositions containing QUE-2, β-Lapachona, and Floretina, as well as the traditional commercial biocides GA and THPS, was evaluated by determining indices that weigh values of uniform corrosion rates (loss of mass), in relation to the respective standard deviations (table previously presented), obtaining the average, minimum and maximum values, to their respective Biological Controls, defining 9 (nine) different comparison possibilities, from the most conservative to the least {

83/113 conservative, for each of the tested BRS cultures. This comparison model, applied to MES (newly formed biofilm), points to a guiding criterion for comparison between different compositions of the present invention and traditional commercial biocides, in the presence of the bacteria, regarding the expectation of uniform corrosion protection insofar as would compute the deviations obtained by determining the corrosion rate by loss of mass, being able to select a more conservative factor, such as, for example, TC (ES) max / TC (CB2) min (ie: ratio established in percentage by maximum corrosion rate in the presence of the biocidal substance, and of the bacterium, evaluated in relation to the minimum corrosion rate in the presence of the bacterium and in the absence of the biocidal substance and in the presence of the DMSO vehicle). In Tables 24 and 25 and Figures 13 A and B for each of the tested cultures, compositions of the present invention and traditional commercial biocides, these comparative criteria are presented. It is noteworthy that the composition containing QUE-2 proved to be, unequivocally, the most efficient in protecting uniform corrosion according to any criterion (more or less, conservative) applied in relation to the two BRS cultures, demonstrating protection capacity (% in relation to the respective Biological Controls in the absence of composition) to corrosion by weight loss between 0.8% and 53.6%, compared to the target culture BRS pool header P-19 and between 25.9% and 51.3% against the culture control BRS Desulfovibrio alaskensis. The composition containing β-Lapachona showed degree of protection according to less conservative indexes in the case of the target culture BRS pool header P-19. The composition containing Floretina and also the traditional commercial biocides have shown a reduced capacity to protect against corrosion by loss of mass, according to less conservative criteria in comparison with the respective Biological Controls.

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TABLE 24: Expectation of uniform corrosion protection for compositions comprising QUE-2, β-Lapachona, Floretina

MES newly formed biofilm CODE Expectation of uniform corrosion protection (SAE 1020 steel biocoupons) against the Antimicrobial Compositions of the present invention in the presence of BRS in seawater (MES - contact 45 days) Mean TC + / SD (mm / year) Minimum CT (mm / year) Maximum CT (mm / year) CORROSION INHIBITION (%) TC (ES) / TC (CB2) - respective culture ES max / CB2 max (%) ES max / CB2 med (%) ES max / CB2 min (%) without med / CB2 max (%) ES med / CB2 med (%) ES med / CB2 min (%) ES min / CB2 max (%) ES min / CB2 med (%) ES min / CB2 min (%) Biological Controls (de-aerated sea water + DMSO) HCB2 0.01018 ± 0.00333 0.00685 0.01351 ALCB2 0.00965 ± 0.00091 0.00874 0.01056 Antimicrobial Compositions in invention HQ 0.00653 ± 0.00026 0.00627 0.00679 49.7 33.3 0.8 51.6 35.9 4.7 53.6 38.4 8.5 ALQ 0.00581 ± 0.00067 0.00514 0.00648 38.6 32.8 25.9 45.0 39.8 33.5 51.3 46.7 41.2 HBL 0.00996 ± 0.00000 0.00996 0.00996 26.3 2.2 -45.4 26.3 2.2 -45.4 26.3 2.2 -45.4 ALBL 0.01176 ± 0.00272 0.00904 0.01448 -37.1 -50.1 -65.7 -11.4 -21.9 -34.6 14.4 6.3 -3.4 HPH 0.01113 ± 0.00132 0.00981 0.01245 7.8 -22.3 -81.8 17.6 -9.3 -83.8 27.4 3.6 -43.2 ALPH 0.01527 ± 0.00235 0.01292 0.01762 -86.9 -82.6 101.6 -44.6 -58.2 -74.7 -22.3 -33.9 -47.8

I ·

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TABLE 25: Expectation of uniform corrosion protection for traditional commercial biocides

MES - newly formed biofilm CODE Expectation of uniform corrosion protection (SAE 1020 steel biocoupons) compared to traditional commercial biocides in the presence of BRS in seawater (MES - contact 45 days) TC (mm / year) average + / sd Minimum CT (mm / year) Maximum CT (mm / year) CORROSION INHIBITION (%) TC (ES) / TC (CB1) - respective culture ES max / CB1 max (%) ES Ί max / CB1 med (%) ES max / CB1 min (%) ES med / CB1 max (%) ES med / CB1 med (%) ES med / CB1 min (%) ES min / CB1 max (%) ES min / CB1 med (%) ES min / CB1 Min (%) Biological Controls (de-aerated sea water) HCB1 0.00728 + - 0.0021 0.00518 0.00938 ALCB1 0.00747 + - 0.00243 0.00504 0.0099 Traditional commercial biocides HGA 0.00758 ± 0.00265 0.00493 0.01023 -9.1 -40.5 -97.5 19.2 -4.1 -46.3 47.4 32.3 4.8 SEAWEED 0.00810 ± 0.00510 0.003 0.0132 -33.3 -76.7 161, 9 18.2 -8.4 -60.7 69.7 59.8 40.5 HT 0.00739 ± 0.00140 0.00599 0.00879 6.3 -20.7 -69.7 21.2 -1.5 -42.7 36.1 17.7 15.6 ALT 0.00935 ± 0.00298 0.00637 0.01233 -24.5 -65.1 144, 6 5.6 -25.2 -85.5 35.7 14.7 26.4

[0244] The example presented shows that compositions of the present invention are efficient in inhibiting planktonic and sessile BRS. In addition, QUE-2 stands out significantly in protecting against uniform corrosion, by controlling the corrosive process by loss of mass.

[0245] Microscopy ultrastructural analysis was also carried out

Transmission Electronics (SEM) of the surface of biocoupons in the presence of newly formed BRS pool header P-19 biofilms after contacts for 18 hours in front of de-aerated sea water (CB1), in front of de-aerated sea water added with the volumetric equivalent of DMSO vehicle of the composition containing QUE-2 (1000 ppm) (CB2), compared to THPS (500 ppm) in de-aerated sea water and compared to the Composition containing QUE-2 (1000 ppm) previously dissolved in DMSO in de-aerated sea water.

[0246] The biocoupons were removed from their contacts for 18 hours and immediately immersed in synthetic and sterile seawater pre-purged with NLO, after which they were fixed for 24 hours at 4 ^o C in a 2.5% glutaraldehyde solution in 0.1M sodium cacodylate buffer, pH 7.2, with 0.14% ruthenium red and 2.5% NaCI. Subsequently, the biocoupons were washed

86/113 three times for 10 minutes each time in 0.1 M sodium cacodylate buffer solution, pH 7.2, with 0.07% ruthenium red and 2.5% NaCI. Then, the biocoupons were post-fixed for 2 h at room temperature in a 1% osmium tetroxide solution in 0.1M sodium cacodylate buffer, pH 7.2, with 0.07% ruthenium red and 2.5% NaCI . After this step, the biocoupons were washed three times for 10 minutes each time in 0.1 M sodium cacodylate buffer solution, pH 7.2, with 0.07% ruthenium red and 2.5% NaCI. Then, the desalination step was carried out in solutions of the same buffer containing decreasing concentrations of NaCI until complete exhaustion. Subsequently, the biocoupons were dehydrated for 15 min in each of the solutions containing increasing concentrations of acetone (30% to 100%). After dehydration, the preserved biocoupons were subjected to subsequent drying steps by the critical point apparatus and metallization with gold. The specimens were observed in the scanning electron microscope Quanta 450 FEG, from the FEI Company.

[0247] The ultrastructural characterization by Electron Microscopy of

Scanning (SEM) of the newly formed BRS pool header P-19 biofilms in carbon steel biocoupons, as previously mentioned, is shown in the photomicrographs of Figures 19 to 22.

[0248] Figure 19 shows a bacterial biofilm covering the surface of the carbon steel biocoupon, recently formed during contact with CB1, with bacteria (black arrows), whose volume and surface are preserved, adhered to the biofilm structure and associated with the formation of FeS. The bacteria have an approximate diameter of 0.3 pm and a length between 1.5 pm and 2.5 pm, which is compatible with the typical dimensions reported for BRS.

[0249] In contact with CB2 - Figure 20 -, bacterial biofilm is observed covering the surface of the carbon steel biocoupon. Countless bacteria (black arrows), whose volume and surface are preserved, are attached to the biofilm structure and associated with the formation of FeS. The bacteria have an approximate diameter of 0.3 pm and a length between 1.5 pm and 2.5 pm, which is compatible with the dimensions

87/113 typical reported for BRS. These observations demonstrate that the DMSO solvent (vehicle) does not cause significant ultrastructural changes.

[0250] After 18 hours of contact with the newly formed biofilm against the commercial biocide THPS (500 ppm) - Figure 21, compared to the control situation CB1 in the presence of de-aerated seawater, it is observed that the bacteria (black arrows) they are in lesser quantity, but present volume, diameter and length still compatible with what is observed in the referred control situation (CB1). It is also observed an association of bacteria with abundant FeS film (white star).

[0251] Figure 22 shows that, compared to the control situation

CB2 in the presence of de-aerated sea water added with DMSO, although the newly formed biofilm in front of the composition containing QUE-2 (1000 ppm) can still be observed covering the surface of the carbon steel biocoupon, it is disorganized, showing a lesser amount of adhered bacteria (black arrows), which are twisted and quite distended, with length (around 5.0 pm) increased by up to two times, compared to the bacteria observed in the control situation (CB2).

[0252] It is evident, therefore, that, even in the short contact time (18 hours), the composition containing QUE-2 showed, compared to THPS, a higher level of effectiveness of antimicrobial activity in the ultrastructural disorganization of the newly biofilm - formed of BRS.

EXAMPLE 10 [0253] Evaluation of the efficiency of the QUE-2 containing composition of the present invention in the culture control of BRS pre-adhered to SAE 1020 carbon steel biocoupons in microcosm consisting of de-aerated sea water using the NLO Sensor Assay Methodology (MES-preformed biofilm) [0254] Biocidal Efficiency was evaluated against the composition containing

QUE-2 (1000 ppm) targeting BRS and anaerobic consortium bacteria (BANHT) originating from the P-19 injection system pool, in sessile form, structured as preformed biofilm on the surface of the SAE 1020 steel biocoupon , and also in planktonic form.

τ

88/113 [0255] The composition containing QUE-2 was selected for the said test due to the good results achieved, previously presented (MES - newly formed biofilm) and indicative of BRS population control in the sessile form and excellent role in the control of the corrosive process by loss of mass in the presence of both tested cultures.

[0256] The BRS header P-19 pool culture maintained in the laboratory, at 10% v / v, in penicillin flask (100 mL) was carried out in the presence of 90.0 mL of modified Postgate C medium. Volume aliquots of 5.0 ml of the said culture, in the growth phase at the age of 18 hours, were transferred to penicillin flasks (50 ml), in a quantity defined for the tests and their respective controls, in the presence of 45.0 ml of modified Postgate C medium where pre-sterilized biocoupons were already disposed, as defined in example 9. The vials already in the presence of modified Postgate C medium, sterile biocoupons and the inoculated culture at a final concentration of 10 ⁷ MPN / mL were incubated for a period of five (5) days to stimulate the formation of the biofilm, which is now observed visually. After this period, a flask with biocoupon for Biological Control was chosen at random, both for planktonic bacteria and for sessile bacteria that might already be attached to the biocoupon (COUTINHO, CMLM, 1991). In the liquid phase, the quantification of planktonic organisms was performed by the NMP technique, in Postgate E medium modified for BRS and by the same technique in BANHT medium for anaerobic bacteria intercropped with BRS.

[0257] The corresponding biocoupon was scraped and the resulting biofilm received in a reducing saline solution (SSR), already as a suspension, which was vortexed, to quantify sessile BRS using the NMP technique, by incubation in Postgate E medium modified. This phase is called pre-formation of the biofilm. After this same period, the remaining pre-filmed biocoupons were removed from the modified Posgate C incubation medium and, in sequence, washed with 9.0 mL of reducing saline and introduced into the 50.0 mL flasks to perform the contact phase. with sea water de-aerated by NLO plus resazurin, whose flasks had previously been identified for each test, which were carried out in

89/113 triplicate for controls CB1 and CB2, and also for tests against the composition containing QUE-2 (concentration 1000 ppm) in DMSO. This phase is called Activation and occurs in the presence of pre-filmed biocoupon.

[0258] In the referred Contact phase, the pre-filmed biocoupons were then incubated for a period of 24 hours in front of the composition containing QUE2, in an oven at a temperature of 30 ° C, after which they were removed, washed with 9 , 0 ml_ of SSR and introduced in penicillin vials with 50.0 ml of Postgate E medium modified to stimulate, and visually reveal, the growth of BRS, in sessile form, by hypothesis still present in each biocoupon. This phase is called Revelation. The liquid phase of the contact step with the biocidal substance for each set of triplicates was pooled, being incubated in Postgate E medium modified according to the NMP technique in modified Postgate E medium and in medium called BANHT, for quantification, respectively, BRS and anaerobic bacteria (BANHT) in consortium, in planktonic form. In addition, a qualitative assay was performed to detect BRS for each triplicate, also by incubation in modified Postgate E medium, at a temperature of 30 C, for a period of up to 28 days.

[0259] The visual observation of the growth indicator due to the possible presence of iron sulfide (blackish color), or its absence, during the Developing phase, follows an experimental procedure to prove the referred visual observation, quantifying BRS present in the liquid phase of the modified Postgate E incubation medium, as planktonic organisms, and as sessile, on the surface of the biocoupon subjected to the Contact with the biocidal substance and Revelation phases already in the presence of the modified Postgate E medium. Briefly, after this phase has ceased (16 days), the aspect of each biocoupon of each triplicate (assay against the biocidal substance and its respective controls) was reported, with each pre-filmed specimen removed and scraped and washing its surface in order to collect the biofilm, possibly present, in the form of a suspension in a defined volume of 10 mL of reducing saline solution (SSR). The suspension is homogenized by vortex, being incubated, according to the same conditions already reported, in Postgate E modified to check the

90/113 presence of bacteria adhered to the biocoupon. This phase is called re-disclosure, already described in a previous example (MES - newly formed biofilm) and aims to carry out a protocol aiming at the quantification of organisms still viable in the preformed biofilm, after stimulating growth in the Developing phase, and , perhaps, resistant to the biocidal substance, when compared to their respective Biological Controls.

[0260] It was observed, after a period of five days, under the conditions previously described as adequate for the growth of mesophilic BRS, the formation of stabilized biofilm on the surface of the biocoupons exposed to the modified Postgate C medium, allowing the performance of the test against the composition of the invention comprising QUE-2 and the traditional commercial biocide THPS with a focus on Biocidal Efficiency relative to BRS in planktonic and sessile forms.

[0261] In Tables 26 and 27 in sequence, we can verify control parameters of the inoculated culture, confirming that it remained active, both in the liquid phase of the growth medium, and in the biocoupon itself, already as a preformed biofilm. , as we can see in the results related to the Revelation phase. A point to note concerns the concentration of the planktonic culture inoculated in a concentration range indicated as conservative (1.4 x 10 ⁷ NMP / mL), which allowed the concentration of 4.5 x 10 ⁴ NMP / mL of scraped biofilm suspension (SSR), equivalent to the concentration of BRS in sessile form at about 4.0 x 10 ⁴ NMP / cm ² relative to the pre-filmed biocoupon area. This culture control parameter is adequate and can be considered conservative for use in the Contact phase in relation to the composition. Additionally, it appears that the anaerobic culture (growth in BANHT medium) combined with BRS follows the same growth trend when evaluated in planktonic form.

[0262] The results related to the evaluation carried out immediately after the contact phase of the biocidal substance with the biofilm, presented in Tables 26 and 27, demonstrate that both the composition containing QUE-2 and the traditional commercial biocide THPS, were capable of control growth in the liquid phase relative to the target crops BRS and BANHT under the

91/113 planktonic form. It is worth noting that the concentration of the referred planktonic targets BRS and BANHT, in their respective controls (CB1) for THPS, reached low values, such as 2.5 x 10 ¹ and 2.5 x 10 ² NMP / mL. Conversely, for the composition containing QUE-2, these respective values (CB2) are 9.5 x 10 ¹ and 2.5 x 10 ¹ NMP / ml. It is noteworthy, therefore, that contact, after 24 hours in front of the two substances, generated non-detection (<0.3 NMP / mL) of planktonic organisms. It is considered that the modified Postgate C medium favored the formation of the biofilm adhered to the surface of the biocoupon to the detriment of the growth of planktonic bacteria in the liquid phase, possibly in solution, those bacterial cells that would be released from it in the Contact phase; considering, this way, that this would constitute the central objective to be reached with the development of the protocol. It is recognized that the referred culture medium, applied to the pre-formation stage of biofilm, is indicated by the classic reference Postgate (1984) as the most suitable for use in research with BRS because its composition allows low generation of solids in solution throughout the experiment due to the presence of sodium citrate chelating agent and low iron content, unlike traditional means applied to detection and isolation, such as Postgate E and B media, which are richer in iron and do not contain said chelating substance in its original formulations. The greater availability in solution of the H ₂ S metabolite, typical of the BRS metabolic action in the anaerobic reduction of sulfates, in the aforementioned medium applied to this protocol, would tend to be better controlled as it would stimulate the deposition of a sulfide film blackened iron (FeS) on the biocoupon surface, which would support the creation of the microenvironment necessary for the propagation of anaerobic bacteria in its sessile form, making the protocol more conservative. Regarding the results related to the Developing phase (Table 27), when the growth by incubation in modified Postgate E medium associated with the surface of the biocoupons after the Contact phase (24 h) is evaluated through visual analysis over time, it was verified It is understood that both the composition containing QUE-2 and the traditional commercial biocide THPS, when compared to their respective Biological Controls, delayed the growth of BRS in sessile form for periods of about 6 to 9 days in the case of the first and 6 to 10 days for the second. This indicative of growth inhibitory action, although

92/113 still does not indicate death of adhered bacterial cells, it allows to compare, among themselves, the effect of the tested substances, in relation to the respective Biological Controls, demonstrating, qualitatively, close bioactivities in relation to the sessile BRS with a view to mitigating its growth process when organized as biofilms.

TABLE 26: Results of evaluation of growth control of planktonic and sessile bacteria performed immediately after the contact phase of the composition comprising QUE-2 and the traditional commercial biocide THPS with the preformed biofilm

MES - preformed biofilm BRS Culture pool header P19 (medium Postgate E mod) concentration (NMP / mL) BANHT culture pool header P-19 (BANHT medium) concentration (NMP / mL) Biological controls of planktonic and sessile culture - Pre-formation phase of biofilm in modified Postgate C medium (time = 5 days) Control of cultures (planktonic BRS and BANHT) inoculated in Postgate C medium in the presence of pre-sterilized SAE 1020 steel biocoupon (time = zero days) 1.4 x 10 ⁷ 4.5 x 10 ⁵ Control of liquid culture (planktonic BRS and BANHT) in the presence of pre-filmed biocoupon (time = 5 days) 4.5 x 10 ⁷ 2.5 x 10 ⁶ Growth control on the surface of the pre-filmed biocoupon (sessile BRS evaluated in liquid phase of the scraped biofilm suspension in SSR) (time = 5 days) 4.5x10 ⁴ (*) NR Biological Controls - Contact Phase (time = 24 hours) CBt - pool of planktonic bacteria in the presence of depleted seawater 2.5 x 10 ¹ 2.5x10 ² CB ₂ - pool of planktonic bacteria in the presence of depleted sea water + DMSO 9.5 x 10 ¹ 2.5 x 10 ¹ Composition containing QUE-2 and traditional commercial biocide THPS - Contact Phase (time = 24 hours) Composition containing QUE-2 1000 ppm <0.3 <0.3 THPS 500 ppm certified <0.3 <0.3

NR- not performed (*) - determination made on the suspension of the biofilm adhered to the biocoupon (scraping the total area and homogenizing in a volume of 10 mL of SSR)

TABLE 27: Results related to the Development phase in the presence of compositions comprising QUE-2 and traditional commercial biocide THPS.

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MES - preformed biofilm Development Phase in Postgate E medium modified from pre-filmed biocoupons after Contact Phase (24 hours) in the presence of the Composition containing QUE-2 and the traditional commercial biocide THPS (”) Biological Controls CBi Positive triplicates in 5 days CB ₂ Positive triplicates in 5 days Composition QUE-2 and traditional commercial biocide THPS Composition containing QUE-2 1000 ppm Positive duplicates in 11 days and all in 14 days THPS 500 ppm certified Positive duplicates in 11 days and all in 15 days

() Information about visual observation (blackening of pre-filmed biocoupon

Grades:

a- results related to the trial ended in 16 days, with positive growth being observed by visual analysis;

b- detection tests by qualitative technique referring to inoculation in triplicate at 10% v / v of the liquid phase of the tests (Contact phase after 24 h) against the Composition containing QUE-2 (1000 ppm) and the traditional commercial biocide THPS ( 500 ppm) in modified Postgate E medium showed growth in both controls over 7 days. In the case of the assay against Composition QUE-2, no change in the color of the medium was observed (not indicating reduction) and, therefore, pointing to biocidal action on the planktonic phase of the experiment. In the case of the substance THPS (500 ppm), a reduction in the medium was observed, which, in this case, may also be associated with the antioxidant activity presented by that substance.

[0263] Aiming at a quantitative analysis that can allow the comparison with control tests and also with the tests against the two

94/113 biocidal substances (Composition containing QUE-2 and traditional commercial biocide THPS), with the aim of evaluating Biocidal Efficiency in defiance of sessile BRS (preformed biofilm), proceeded to the Revelation, as briefly described previously (newly formed biofilm). The results, commented on below, are shown in Table 28 and Figure 14.

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TABLE 28: Results related to the re-development phase in the presence of compositions comprising QUE-2 and traditional commercial biocide THPS.

MES - preformed biofilm - Re-Development Phase Biocidal efficiency in challenge to planktonic and sessile BRS cultures Culture BRS pool header P19 - planktonic bacteria present in the liquid phase of the modified Postgate E medium Culture BRS pool header P-19 - sessile bacteria present in the suspension of the biofilm adhered to the biocoupon in modified Postgate E medium Controls Biological Concentration (NMP / mL) CBj {pool) 1.1 x 10 ⁵ 4.5 x 10 ⁴ CB ₂ {pool) 4.5x10 ’ 1.1 x 10 ’ Composition QUE-2 and commercial biocide THPS BRS concentration (NMP / mL) (Biocidal efficiency%) Composition containing QUE-2 1000 ppm Disclosure 1 - liquid phase 0.9 x 10 ’(> 99.99%) Revelation 2 - liquid phase 0.9 x 10 ¹ (> 99.99%) Development 3 - 2.5 x 10 'liquid phase (> 99.99%) Revelation 1 (*) - biocoupon <0.3 (100%) Revelation 2 (*) - biocoupon 0.9 x 10 ¹ (99.99%) Revelation 3 (*) - biocoupon <0.3 (100%) THPS certification 500 ppm Revelation 1 '- liquid phase 1.4 x 10 ⁴ (87.27%) Revelation 2 '- liquid phase 1.4 x10 ⁴ (87.27%) Revelation 3 '- liquid phase 2.5 x 10 ³ (97.73%) Revelation 1 '(*) -biocoupon 4.5 x 10 ² (99.00%) Revelation 2 '(*) - 4.5 x 10 ² biocoupon (99.00%) Revelation 3 '(*) - biocoupon 2.5 x 10 ² (99.44%)

(*) determination made in the suspension of the biofilm adhered to each biocoupon (scraping the exposed area, followed by homogenization in a volume of 10 mL of SSR) of each set of experiments in triplicate.

[0264] Through a purely comparative analysis of the results obtained, from the re-Disclosure applied to the Methodology of the Sensing Tests (MES), it becomes possible to demonstrate, through the determination of the Biocidal Efficiency (see table presented above) achieved in challenge to Planktonic and sessile BRS (preformed biofilm) originating from P-19 pool header culture, that the composition containing QUE-2 (1000 ppm) showed biocidal activity significantly superior to that presented by the traditional commercial biocide THPS (500 ppm) (Figure 15).

[0265] It should be noted that the Biocidal Efficiency results achieved in relation to the composition containing QUE-2 are relevant in

96/113 in relation to those previously presented, which are related to the application of the Sensor Assay Methodology (MES) in the presence of newly formed biofilm, when the traditional commercial biocide THPS and GA showed the same death efficiency demonstrated compared to the composition containing QUE- 2. In the experiment in the presence of preformed biofilm, a condition that can be considered more conservative for the oil industry, the composition of the invention comprising QUE-2 also presents itself as an alternative of interest for controlling uniform corrosion in the presence of BRS in microcosm consisting of de-aerated sea water. A relevant comparative advantage was identified for the composition containing QUE-2 (1000 ppm) over the traditional commercial biocide THPS (500 ppm).

[0266] It should be clarified that the concentration of the composition containing

QUE-2 observes the same range used in the case of the traditional commercial biocide THPS, under conditions similar to those in the example, as shown in the following reference (http://msdssearch.dow.com/PublishedLiteratureDOWCOM/dh_030b/0901b803 8030b48f.pdf? Filepath? = biocides / pdfs / noreg / 253-01944.pdf & fromPage = GetDoc). [0267] The microbiological evaluation of the composition containing QUE-2 (1000 ppm) in defiance of the preformed BRS biofilm in carbon steel biocupons was integrated with the ultrastructural evaluation (assays and respective controls) by Scanning Electron Microscopy (SEM). ) with specimens preserved for analysis. Such evaluation included, initially, immersion of carbon steel biocoupons, pickled and pre-sterilized by electron beam (E-beam), in the presence of a concentrated BRS pool header P-19 culture (10 ⁸ bacteria / mL) in Postgate C medium modified for a period of five days at 30 ° C, after which they were removed from the medium, washed in sterile reducing saline (SSR) in a vertical laminar flow hood under constant purging of oxygen-free nitrogen (NLO). Biocoupons resulted in preformed biofilms, which were then placed in contact for 24 h with the composition containing QUE-2 (1000 ppm) in microcosm consisting of de-aerated seawater for antimicrobial bioactivity tests in defiance of the preformed biofilm of BRS, in relation to the respective biological controls and also comparing to the biocide

97/113 commercial THPS (500 ppm). In parallel to these procedures for evaluating the growth of bacteria by microbiological techniques, which demonstrate the effectiveness of the said composition, four biocoupons were separated and, after 24 hours, removed from their respective test solutions (contacts), specified below, for analysis by Scanning Electron Microscopy (SEM): 1) CB1 - Biological Control 1 - contact in sterile seawater; 2) CB2 - Biological Control 2 - contact with sea water added with the solvent (vehicle) DMSO in a concentration equivalent to that used to prepare the composition containing QUE-2 (1000 ppm); 3) contact in sea water in the presence of the traditional commercial biocide THPS (500 ppm); 4) contact in sea water in the presence of the composition containing QUE-2 (1000 ppm). All tests were performed at a temperature of 30 ° C.

[0268] Immediately after withdrawal, said biofouled biocoupons were immersed in synthetic and sterile seawater pre-purged with NLO followed by preservation by fixing for 24 hours at 4 ° C in 2.5% glutaraldehyde solution. 0.1 M sodium cacodylate buffer, pH 7.2, with 0.14% ruthenium red and 2.5% NaCI. After this step, they were washed three times for 10 minutes each time in 0.1 M sodium cacodylate buffer solution, pH 7.2, with 0.07% ruthenium red and 2.5% NaCI. Then, the desalination step was carried out in solutions of the same buffer containing decreasing concentrations of NaCI until complete exhaustion. Subsequently, the biocoupons were dehydrated for 15 min in each of the solutions containing increasing concentrations of acetone (30% to 100%). After dehydration, the preserved biocoupons were subjected to subsequent drying steps by the critical point apparatus and metallization with gold. The samples were observed in the Quanta Inspect Scanning Electron Microscope.

[0269] The ultrastructural characterization by Electron Microscopy of

Scanning (SEM) of BRS pool header P-19 biofilms preformed in carbon steel biocoupons, previously mentioned, is shown in the photomicrographs of Figures 23 to 25.

[0270] Figure 23 shows a typical bacterial biofilm covering the carbon steel biocoupon surface after 24 hours of contact with the

98/113 biological control CB2. Countless bacteria (white arrows), whose volume and surface are well preserved, are attached to the structure of the biofilm, associated with microformations of FeS (white star) and other compounds. The bacteria have an approximate diameter of 0.3 pm and a length between 1.5 and 2.0 pm, attesting to typical BRS dimensions. This evidence indicates that the DMSO solvent (vehicle) does not cause significant ultrastructural changes in the preformed BRS biofilm.

[0271] Already in Figure 24, it is observed in preformed biofilm, treated with commercial biocide THPS (500 ppm) for 24 hours of contact, that the bacteria (white arrows), compared to the control situation CB1 in the presence of water from the deserted sea, they are less frequent, but still show dimensions, such as volume, diameter and length (~ 2.0 pm), compatible and typical of BRS.

[0272] In Figure 25, the preformed biofilm, treated for 24 hours with the Composition containing QUE-2 (1000 ppm), shows a smaller amount of adhered bacteria (white arrows), compared to what is observed in the biological control CB2 in the presence of de-aerated sea water with DMSO, which presents with altered morphology, very rough and distended surface, with more than twice the length (around 4.5 pm) typical of BRS.

[0273] In short, the photomicrographs demonstrate, in the respective control CB2, the preservation of the preformed biofilm and associated with the surface of the carbon steel biocoupon, as well as the disorganization of the preformed biofilms when in the presence of the mentioned substances endowed with characteristics biocides, highlighting the greater effectiveness of the Composition containing QUE-2 (1000 ppm) compared to THPS (500 ppm).

EXAMPLE 11 [0274] Evaluation of Antimicrobial Activity of Compositions

Antimicrobials of the invention comprising Butein, Lapachol and βLapachona against microorganisms, particularly strains of environmental bacteria and health bacteria.

[0275] The target microorganisms selected for the assays against the compositions of the invention included facultative aerobic bacteria (Gram

99/113 positive and Gram-negative). In the first list of target bacteria, which is classified as from the environmental area, two strains originate from the oil industry, forming part of the Petrobras / Cenpes Collection, which were isolated and identified (SEBASTIÁN, GV, 1999), from testimony of deepwater reservoir in Campos Basin, RJ, Brazil: Bacillus licheniformis T3.1 and Bacillus licheniformis T3.2. The other bacterial strains originate from the American Type Culture Collection (ATCC) and National Type Culture Collection (NTCC), belonging to the Collection of Cultures of the Genus Bacillus and Correlates (CCGB), of the Laboratory of Bacterial Physiology (LFB) / Fiocruz:

- Bacillus amyloliquefasciens CCGB 452 (LFB-FIOCRUZ 452) - ATCC 23892

- Bacillus licheniformis CCGB 125 (LFB-FIOCRUZ 125) - ATCC 14580

- Bacillus subtilis CCGB 732 (LFB-FIOCRUZ 732) - ATCC 6633

- Bacillus cereus CCGB 406 (LFB-FIOCRUZ 406) - NTCC 2599

- Paenibacillus macerans CCGB 126 (LFB-FIOCRUZ 126) - ATCC 8244 [0276] In the second list of target bacteria, which is typical of the health area, all the strains tested come from the Collection of Bacterial Cultures of the National Institute of Control of Quality in Health (INCQS / Fiocruz), preserved and preserved by the LFB / Fiocruz:

- S. aureus ATCC 25923

- E. coli'ATCC 25922

- E. faecalis ATCC 29212

- P. aeruginosa ATCC 278530

- S. Enteritid is ATCC 13076

- S. Typhi ATCC 19214

- S. flexneri ATCC 12022 [0277] In a first step, the antimicrobial activity of the compositions of the present invention was evaluated by the Disk Diffusion Assay in Müeller-Hinton medium (physiological salinity culture medium) and / or, in sequence, these pre-selected compositions were evaluated using the CMB technique. The tests performed are briefly described below.

[0278] The inoculum concentration of the bacterial culture has been adjusted to

100/113 adequate value (1 x 10 ⁸ UFC / mL) for the next stage of disk diffusion, observing the standard defined by the Clinicai and Laboratory Standards Institute (CLSI, 2014; http://clsi.org/standards/micro /). In this case, the bacterial strain is grown under conditions such as to achieve the standard turbidity of the Mc Farland Scale 0.5 tube and through a UV-visible spectrophotometer (reading from 0.08 to 0.10 at absorbance of 625 nm). adjustment of the inoculum with sterile physiological saline.

[0279] Cultures of the bacteria selected at the above concentration were transferred to Müeller-Hinton Agar plates. Filter paper discs (Whatman AA Discs, 6.0 mm) impregnated with 10 pL of the respective composition of the invention after drying at 33 ° C (CLSI, 2014) were placed on the agar surface, the plates were closed and maintained for 18 -24h in an oven at 37 ° C. Then, each of these plates was observed for the formation of a bacterial growth inhibition halo around the paper discs, halos resulting from the antimicrobial bioactivity of the compositions of the invention. Qualitatively identified the antimicrobial activity, the CMB was determined for the compositions of the invention, as reported below.

[0280] The inoculum for each culture was adjusted to a bacterial concentration of 1 x 10 ⁸ CFU / mL and then diluted 1:10 in 9mL of Broth Müeller-Hinton to 1 x 10 ⁷ CFU / mL .

[0281] The tests were performed on a Microdilution Plate. To each of the wells of this plate containing 100pL of Müeller-Hinton broth, 100pL of the composition of the invention were added in the different concentrations to be tested (serial dilutions), followed by the addition of 5pL of the diluted inoculum (bacterial concentration of 1 x 10 ⁷ CFU / mL).

[0282] The preparation of dilutions of the compositions of the invention follows the CLSI standard, comprising final concentrations in the range of 512 pL to 0.25 pL. The assay includes controls containing Broth Müeller-Hinton, Broth MüellerHinton plus each of the compositions of the invention (11th well) and Broth Müeller-Hinton plus adjusted inoculum of the strains tested. After inoculation, the Microdilution Plate was kept in a 37 ° C oven for 18-24h. After this time, the final bacterial concentration was 5 x 10 ⁵ CFU / mL. It was done,

101/113 then, transferring a 10 pL aliquot from the first well, which did not show turbidity, to a test tube containing 10 ml_ of Broth Müeller-Hinton. This dilution operation was repeated for the other wells without turbidity. The final bacterial concentration obtained corresponded to 5 x 10 ² CFU / mL. An aliquot of 0.1 ml was removed from each of these dilutions for dispensing on a Blood Agar plate for the determination of CMB. In the example that follows and in the others in sequence, the protocol described above was adopted.

[0283] The results of the challenges of compositions containing Butein,

Lapachol and β-Lapachona of the present invention against strains of environmental bacteria and human and animal health, according to the tests described above, are shown in Table 29 below. The composition containing β-Lapachona showed strong evidence of inhibition by the Disk Diffusion assay, being, in sequence, subjected to the assay to determine the CMB.

TABLE 29: Results of challenges of compositions comprising

Butein, Lapachol, β-Lapachona against different bacterial strains

Target bacteria Butein Lapachol β-Lapachona β-Lapachona CMB (ppm) Strains of environmental bacteria Inhibition halos (diameter CMB Bacillus amyloliquefasciens * 6mm 12mm 20mm Bacillus licheniformis Cenpes 3.1 * 0 6 / 8mm 20mm £ 36.7 Bacillus licheniformis Cenpes 3.2 * 0 7mm 21mm Bacillus licheniformis 0 11mm 22mm Paenibacillus macerans 0 7mm 25mm Bacillus subtilis * 6mm 7 / 9mm 19mm Bacillus cereus 6 / 8mm 20mm £ 4.6 Strains of health bacteria Staphylococcus aureus 8mm 15mm 21mm £ 73.4 Escherichia coli 0 0 0 Enterococcus faecalis 6mm 0 18mm Pseudomonas aeruginosa 0 0 0 Salmonella Enteritidis 0 0 0 Salmonella Typhi 0 0 0 Shigella flexnerí 0 0 0

[0284] Table 29 shows that there was the formation of an inhibition halo indicative of the antimicrobial bioactivity of the compositions of the invention against

102/113 certain strains of environmental bacteria and health bacteria. It is worth mentioning the bioactivity, expressed by the low CMB value of the composition containing β-Lapachona of the invention against Bacillus cereus.

EXAMPLE 12 [0285] Evaluation of the antimicrobial activity of the compositions of the invention comprising QUE-2, β-Lapachona and Floretina against microorganisms, particularly strains of environmental bacteria and health bacteria by determining CMB.

[0286] Tables 30 to 32 show the antimicrobial activity of the said Compositions of the invention against a wide spectrum of strains of health and environmental bacteria.

TABLE 30: Evaluation of the antimicrobial activity of the composition comprising QUE-2 against different bacterial strains by determining the CMB.

Target bacteria QUE-2 Strains of environmental bacteria Inhibition halos (diameter) CMB (ppm) Bacillus amyloliquefasciens 13 mm 10.04 Bacillus licheniformis T3.1 Cenpes 11 mm 20.08 Bacillus licheniformis T3.2 Cenpes 10 mm 20.08 Bacillus licheniformis 10 mm 20.08 Paenibacillus macerans 7 mm 20.08 Bacilllus cereus 10 mm 10.04 Bacillus subtilis 11 mm 10.04 Strains of health bacteria Inhibition halos (diameter) CMB (ppm) Pseudomonas aeruginosa X 321.26 Salmonella Typhi X 642.53 Salmonella Enteritidis X 642.53 Shigella flexneri X 642.53 Enterococcus faecalis X 80.32 Staphylococcus aureus X 20.08 Escherichia coli X 321.26

X - test not performed

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TABLE 31: Evaluation of the antimicrobial activity of the composition comprising β-Lapachona against different bacterial strains by determining the CMB.

Target bacteria β-Lapachona Bacterial strains environmental Inhibition halos (diameter) CMB (ppm) Bacillus amyloliquefasciens 14 mm 2.52 Bacillus licheniformis T3.1 Cenpes 12 mm 10.1 Bacillus licheniformis T3.2 Cenpes 15 mm 2.52 Bacillus licheniformis 17 mm 1.26 Paenibacillus macerans 9 mm 10.1 Bacilllus cereus 13 mm 10.1 Bacillus subtilis 13 mm 2.52 Strains of health bacteria Inhibition halos (diameter) CMB (ppm) Pseudomonas aeruginosa 0 mm 1292.5 Salmonella Typhi 0 mm 1292.5 Salmonella Enteritidis 0 mm 1292.5 Shigella flexneri 0 mm 1292.5 Enterococcus faecalis 12 mm 80.78 Staphylococcus aureus 17 mm 20.2 Escherichia coli 0 mm 1292.5

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TABLE 32: Evaluation of the antimicrobial activity of the composition comprising Floretine against different bacterial strains by determining the CMB.

Target bacteria Floretina Strains of environmental bacteria CMB (ppm) Bacillus amyloliquefasciens 24.6 Bacillus licheniformis T3.1 Cenpes 3.07 Bacillus licheniformis T3.2 Cenpes 6.15 Bacillus licheniformis 1.54 Paenibacillus macerans 12.3 Bacilllus cereus 3.07 Bacillus subtilis 6.15 Strains of health bacteria CMB (ppm) Pseudomonas aeruginosa X Salmonella Typhi 1173.03 Salmonella Enteritidis X Shigella flexneri X Enterococcus faecalis 6.0 Staphylococcus aureus 12.0 Escherichia coli 2346.05

X- test not performed [0287] The set of tests performed against the compositions containing QUE-2, Floretina and β-Lapachona, according to the protocol previously described in challenge to the strains of health and environmental bacteria also already reported, show results , according to the tables above, which show the efficiency and specificity of the referred compositions against the target bacteria, differing from the state of the art in most cases. In this case, the composition containing QUE-2 stands out, since its bioactivity against all tested cultures is unknown to the state of the art. It should be noted that the compositions containing β-Lapachona and Floretina also differ from the state of the art, in view of the bioactivity against most of the cultures tested and, in some cases, this bioactivity is predicted in the state of the art for the substances that make up the composition of the

105/113 invention.

[0288] The bioactivity of the compositions containing QUE-2, Floretina and β-Lapachona, is summarized, in general in Table 33, presenting the respective CMB ranges.

TABLE 33: Bioactivity of the compositions comprising QUE-2, Floretina and β-Lapachona.

Biocidal Compositions Strains of inhibited environmental bacteria CMB (ppm) Strains of inhibited health bacteria CMB (ppm) QUE-2 7 10.04-20.08 7 20.08-642.53 Floretina 7 3.07 - 24.6 4 * 6.0-2346.05 β-Lapachona 7 1.26-10.1 7 20.2-1292.5

* 4 strains were tested in the reported inhibition range [0289] In Table 34, the most outstanding results are presented compared to the compositions containing β-Lapachona, Floretina and QUE-2, according to the CMB determination protocol, with their respective ranges .

TABLE 34: Bioactivity of the compositions comprising QUE-2, Floretina and β-Lapachona

Biocidal Compositions Strains of environmental bacteria CMB (ppm) Strains of health bacteria CMB (ppm) QUE-2 10.04-20.08 20.08-80.32 * Floretina 3.07 - 24.6 12.0-6.0 * β-Lapachona 1.26-10.1 20.2-80.78 *

* inhibition of Staphylococcus aureus and Enterococcus faecalis [0290] The results presented in the previous Tables show as specific characteristics of the compositions of the invention significant bioactivity over a wide spectrum of strains of environmental bacteria and of human and animal health bacteria, in view of the performance evidenced by the respective CMBs in low concentrations, which bring them closer to substances with antibiotic characteristics. This being a relevant data for the Antimicrobial Compositions of the invention when looking for toxicity and cost reduction.

106/113 [0291] Table 35 summarizes the bioactivity of particular compositions of the invention for inhibiting the growth of strains of environmental bacteria and health bacteria.

TABLE 35: Bioactivity of compositions comprising QUE-2, βLapachona and Floretina against different strains of environmental bacteria and also with an impact on health.

Bacterial strains environmental Antimicrobial Compositions QUE-2 CMB (ppm) β- Lapachona CMB (PPm) CMB Floretine (ppm) Bacillus amyloliquefasciens 10.04 2.52 24.6 Bacillus licheniformis T3.1 Cenpes 20.08 10.1 3.07 Bacillus licheniformis T3.2 Cenpes 20.08 2.52 6.15 Bacillus licheniformis 20.08 1.26 1.54 Paenibacillus macerans 20.08 10.1 12.3 Bacilllus cereus 10.04 10.1 3.07 Bacillus subtilis 10.04 2.52 6.15

[0292] We highlight an important application of these compositions of the invention against the target Bacillus cereus, in view of the reported capacity for said microorganism to degrade aliphatic and aromatic hydrocarbons present in corrosion inhibitors influencing corrosion in diesel transport ducts. It is also worth mentioning the inhibition of two strains of environmental bacteria in the petroleum area, Bacillus licheniformis T 3.1 and Bacillus licheniformis T 3.2.

[0293] Another aspect to be highlighted is the bioactivity evidenced by the referred compositions of the present invention against all seven strains of Gram-positive environmental bacteria of the genus Bacillus that were the target of the evaluation, when all these microorganisms already show indications of resistance to traditional antimicrobials. normally available, such as

107/113 can be seen in the antibiogram in Table 36 below. In order of relevance, the bioactivity of compositions containing β-Lapachona, Floretina and QUE-2 with CMBs varying, respectively, from 1.26 to 10.1 ppm, from 1.54 to 24.6 ppm and from 10.04 to 20.08 ppm. This result is of great importance for the constitution of compositions destined for the environment, for asepsis, disinfection and / or sterilization, in domestic, private, public and industrial services, including the oil industry, and as alternatives to reduce hospital infections. .

[0294] The composition of the present invention shows, as substances with antibiotic characteristics, significant bioactivity at low concentrations expressed in the respective CMBs. These low levels demonstrate the effectiveness and specificity in the bactericidal action against a wide spectrum of bacteria of the genus Bacillus. It should be noted that strains of environmental bacteria also have an impact on health, as in the case of Bacillus cereus, for example, which causes food poisoning similar to that caused by Staphylococcus sp. The resistance of these Bacillus bacteria to certain traditional antimicrobials is shown in the antibiogram in Table 36.

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TABLE 36: Antibiogram showing the resistance of bacteria of the genus Bacillus to certain traditional antimicrobials.

Antibiogram Multi-resistance profile and susceptibility to traditional antimicrobials presented by strains of environmental bacteria \ Strains Antimicrobials \ / Concentration \ B. licheniformis T3.1 B- licheniformis T3.2 B. subtilis B. licheniformis B. amyloliquefasciens B. cereus Paenibacillus macerans ATCC 8244 Nalidixic Acid (AN) - 30 pg s R s s s R s Ampicillin (AP) - 10 μ g R R R s s R s Bacitracin (BC) - 0.04U R R s R R R R Kanamycin (CN) - 30 μ g s R s s s s s Cephalothin (CF) - 30 μ g R R R s s R s Cefoxitin (CT) - 30 μ g R R R s s R s Clindamycin (CL) - 2 μ g R s s R s R s Chloramphenicol (CO) - 30 μ g R s s s s s s Erythromycin (ER) -15 μ g s R s s s s R Streptomycin (ET) -10 μ g s s s s s s R Phosphomycin (FO) - 200 μ g R R s R s R R Gentamicin (GM) -10 μ g R s s s s s s Lincomycin (LN) - 2 μ g R R R R R R R Polymyxin B (PL) - 300U R R R s R R s Penicillin G (PN) - 10U R R R s s R s Rifampicin (RFM) - 5 μ g R R R s s R s Sulfonamide (SF) - 300 μ g s s s s s s s Sulfazotrin (SXT) - 1.25 / 23.75 μ g R R s s s R s Tetracycline (TT) - 30 μ g s s s s s R s

The interpretation of the inhibition halos was performed according to the criteria of the

Clinical and Laboratory Standards Institute (2014), for antimicrobials in

109/113 overall. The standard sample S. aureus ATCC 25923 was used as control of the assay. R - resistance; S - sensitivity [0295] Table 37 summarizes the bioactivity of Compositions

Particular antimicrobials of the present invention against strains of health bacteria.

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TABLE 37: Bioactivity of Antimicrobial Compositions of the invention comprising QUE-2, β-Lapachona and Floretine against bacterial pathogenic targets.

Strains of health bacteria Antmicrobial Compositions QUE-2 CMB (ppm) β- Lapachona CMB (ppm) CMB Floretine (ppm) Pseudomonas aeruginosa 321.26 1292.5 X Salmonella Typhi 642.53 1292.5 1173.03 Salmonella Enteritidis 642.53 1292.5 X Shigella flexneri 642.53 1292.5 X Enterococcus faecalis 80.32 80.78 6.0 Staphylococcus aureus 20.08 20.2 12.0 Escherichia coli 321.26 1292.5 2346.05

X- test not performed [0296] High efficacy of the composition containing Floretine is observed in defiance of the target bacteria Enterococcus faecalis and Staphylococcus aureus evidenced by the respective CMBs of 6.0 ppm and 12.0 ppm. As well as the composition containing QUE-2 in defiance of the target bacteria Staphylococcus aureus and Enterococcus faecalis, it has CMBs of 20.08 ppm and 80.32 ppm, respectively. The composition containing β-Lapachona presented a CMB of 80.78 ppm in defiance of the target bacteria Enterococcus faecalis. Another important characteristic is the broad spectrum of target bacteria, Gram-negative, with emphasis on Gram-positive bacteria, which proved to be susceptible to the referred compositions. In order of relevance, the bioactivity of the compositions containing QUE-2 (seven strains), β-Lapachona (7 strains) and Floretina (4 strains) with CMBs varying from 20.08 to 642.53 ppm, respectively. 20.2 to 1292.5 ppm and from 6.0 to 2346.05 ppm.

[0297] Therefore, the compositions of the invention are important alternatives to traditional antimicrobials for the treatment of bacterial infectious processes typical of human and / or animal health, and may act

111/113 as drugs also, for example, for topical use. These results, together with the others obtained, constitute particularities of the present invention in relation to the state of the art.

EXAMPLE 13 [0298] Determination of the antioxidant activity of Compositions

Particular antimicrobials of the invention [0299] Evaluation of the antioxidant activity, by the DPPH method, of the Antimicrobial Compositions of the present invention comprising substances of the flavonoid class, and of the monomeric dihydrochalcone category, Floretine and QUE-2, in the presence of the DMSO vehicle. These compositions were selected for the test because they perform well as biocides, against BRS cultures in the oil industry, and also against strains of environmental bacteria and health bacteria, as shown in the previous examples.

[0300] The DPPH method consists of a reduction test, under standardized conditions, of the stable free radical 2,2 diphenyl-1-picrylhydrazyl (DPPH), for the determination of the antioxidant activity (AO) of substances potentially donating hydrogen. The DPPH radical has a very intense absorption in the visible spectrum region (517 nm), which can be easily determined by spectrophotometry until exhaustion, thus attesting to its reduction [MOLYNEUX, P. “The use of the stable free radical diphenylpicrylhydrazyl ( DPPH) for estimating antioxidant activity ”. Songklanakarin J. I know. Technol. Vol. 26 (2): 211-219. 2004.

http://rdo.psu.ac.th/sjstweb/journal/26-2/07-DPPH].

[0301] The tested substance has the potential to act as an antioxidant, through the inhibition or reduction of free radicals, such as DPPH, which is determined by the concentration equivalent to 50% sequestration of the DPPH radical (EC ₅ o) or that is, the concentration at which there is a 50% reduction in the characteristic absorbance of the DPPH radical compared to the white test.

[0302] Initially, a screening was carried out for preliminary comparison of the antioxidant activity (AO) of the said compositions of the invention tested each in three concentrations in DMSO (140 ppm, 70

112/113 ppm, 35 ppm), relative to their respective controls in the absence of substances. In each of these concentrations of the tested compositions, the consumption (sequestration) of the free radical DPPH (100 μΜ in ethanol) was monitored, by decreasing its absorbance, at a wavelength of 517 nm, which was measured in a UV-spectrophotometer. Vis Spectra Max Plus 394 Molecular Devices. In this step, it became possible to compare the compositions with each other, pre-defining, even more accurately, the concentration range to be applied in the tests that determine the EC50 parameter. In sequence, already in a more adjusted concentration range, the sequestration equivalence of the referred radical in the presence of each composition is estimated from obtaining absorbance readings at 517 nm for each concentration, obtaining a decreasing curve with 0 increase concentration, defining, by computational or graphic method, the parameter EC50. As each of the compositions has DMSO as its vehicle, they have been compared (respective volumetric equivalents) with the control of this vehicle in terms of DPPH sequestration. Ascorbic acid was evaluated as a standard substance of recognized antioxidant activity (AO), characterized as a positive control.

[0303] All experiments were performed in triplicate, and the results were calculated in terms of means and standard deviations (sd), being graphically expressed as means (all experiments showed deviations less than 10% from the calculated average). The reagents applied in this protocol were from Sigma, or equivalent.

[0304] The EC50 parameter θ estimated graphically or from an equation of the analytical curve elaborated according to regression analysis (determination of the correlation coefficient r) using the EXCEL program. The determination of EC ₅₀ makes it possible to compare the antioxidant activity (AO) of the compositions with each other, in relation to the reduction of the DPPH radical and with patterns of reactivity known with DPPH, as is the case of ascorbic acid (Molyneux, 2004). The results for compositions containing Floretine and QUE-2 show strong evidence of antioxidant activity as measured by the consumption (sequestration) of the DPPH radical.

113/113 [0305] It is observed that the compositions containing Floretina and QUE-2 show a strong tendency to susceptibility to the DPPH radical, promoting its reduction already in low concentrations, determining for the composition containing QUE-2, EC ₅₀ of 20.5 ppm, and for composition containing Floretine, EC50 of 13.2 ppm. These results are certified by the test against the reference standard of ascorbic acid, with EC ₅₀ of 4.29 ppm, compatible with the reference of the classic literature mentioned above (Molyneux, 2004), which reports EC ₅₀ equivalent to 5 .00 ppm for this standard substance.

[0306] The results obtained against the compositions containing QUE-2 and Floretina of the present invention stand out for the importance that the antioxidant property presents when combined with the antimicrobial bioactivity of the compositions in the industrial area, particularly in the oil industry, when looking for , in particular, non-oxidizing biocides. In the health area, compositions made up of these antioxidant molecules play a relevant role in acting on inflammation, infection, diabetes, nutraceuticals, antitumor activity, sun protection, food preservation, emphasizing, also, their potential use in foods and drinks to replace undesirable antioxidant substances.

[0307] Although several embodiments that incorporate the teachings of the present invention have been shown and described in detail here, those skilled in the art will be able to readily see many other varied embodiments that may still incorporate these teachings. Any equivalent aspects are intended to be within the scope of this invention. In fact, various modifications of the invention in addition to those shown and described here will become apparent to those skilled in the art from the preceding description. Such modifications are also intended to be within the scope of the appended claims.

Claims (31)

1. Antimicrobial composition, characterized by comprising: a) 1% to 98% (weight / volume) of at least one bioactive compound selected from a compound of formula (I) and / or a compound of formula (IIA) and / or a compound of formula (IIB): Formula (I) in which the respective substituents can be selected from the groups: Rt-OH and R ₂ -H; OH; OCH ₃ and R ₃ -H; OH and 2/6 R ₄ - H; OH and R ₅ -H; OH; OCH ₃ R’- R ₆ -H; OH; CH ₃ ; Cl; Br R ₇ - H; OH; OCH ₃ ; NO ₂ ; SO ₃ H Rs - OH; OCH ₃ ; F; Cl; Br; CH ₃ ; NH ₂ ; NO ₂ ; SO ₃ H; C ₄ H ₉ NH ₂ ; OC ₂ H ₅ NH _2; OCH ₂ O Rg-H; OH; OCH ₃ ; OCH ₂ O R10-H; OH Formula (IIA) Formula (IIB) Rn - OH; R1 ₂ -H; OH b) acceptable vehicle; and c) optionally one or more adjuvants. Composition according to claim 1, characterized in that the compound of formula (I) is a phenolic skeletal compound of the flavonoid class 3/6 preferably selected from chalcones, dihydrochalcones and dimeric dihydrochalcones. Composition according to claim 2, characterized in that the compound of formula (I) preferably comprises dihydrochalcones selected from 3- (4-hydroxyphenyl) -1- (2,4,6-trihydroxyphenyl) -1-propanone (Floretine) and 3- (4methoxyphenyl) -1- (2,4,6-trihydroxyphenyl) -1-propanone (QUE-2), dimeric dihydrochalcones such as 1.1 '- (2.2', 4.4 ', 6, 6'-hexahydroxy [1,1 '-biphenyl] -3,3'-diyl) bis [3- (4methoxyphenyl) -1-propanone (Dihydrochalcone dimer-1) and chalcones selected from 2', 3,4,4 '-tetrahydroxichalcona (butein) and 2 ^ 4.4 ^ 6 tetrahydroxichalcona (chalcone naringenin). Composition according to claim 1, characterized in that the compound of formula (IIA) and the compound of formula (IIB) are compounds of the phenolic skeleton of the naphthoquinone class. Composition according to claim 4, characterized in that the compound of formula (IIA) is preferably 2-hydroxy-3- (3-methyl-2-butenyl) -1,4naphthalenedione (Lapachol) and the compound of formula ( IIB) be 2,2-dimethyl-3,4-dihydro-2H-benzo [h] chromene-5,6-dione (β-Lapachona). Composition according to any one of Claims 1 to 5, characterized in that the adjuvant is selected from chelating and surfactant agents, corrosion inhibiting substances, solid supports, carriers of active substances, membrane permeabilizers, preservatives, and / or inhibitors metabolites of sulfate-reducing bacteria (BRS). Composition according to claim 6, characterized in that the chelating and surfactant agents are ethylenediamino tetraacetic acid (EDTA), ethylenediamine diacysuccinate (EDDS) and molecular structures of the chemical category of saponins. Composition according to claim 6, characterized in that the corrosion inhibiting substances are selected from capsaicin and its analogs. Composition according to claim 6, characterized in that the solid supports are selected from silica, polymers derived from silica and polypropylene. 4/6 Composition according to claim 6, characterized in that the carriers of the active substances are selected from cyclodextrins and derivatives. Composition according to claim 6, characterized in that the metabolic inhibitors of BRS are selected from selenite, selenate, monofluorophosphate, molybdate, nitrite, nitrate and their associations. Composition according to any one of claims 1 to 11, characterized in that it is for use in the prevention, mitigation and control of biogenic processes. Composition according to claim 12, characterized in that at least one bioactive compound is comprised in concentrations in the range of 10 to 100,000 ppm, preferably from 10 to 5,000 ppm, more preferably from 50 to 1,000 ppm. Composition according to claim 12 or 13, characterized in that the acceptable vehicle comprises one or more vehicles selected from DMSO; sea water; fresh water; brackish water; isopropanol; glycerol and / or other higher alcohols, such as propylene glycol; sodium chloride and / or other salts. Composition according to any one of claims 1 to 11, characterized in that it is for use in inhibiting the growth of strains of environmental bacteria. Composition according to claim 15, characterized in that at least one bioactive compound is comprised in concentrations in the range of 1 to 10,000 ppm, preferably from 1 to 50 ppm, more preferably from 1 to 30 ppm. Composition according to claim 15 or 16, characterized in that the acceptable vehicle comprises one or more vehicles selected from DMSO; low salinity natural water; sea water; fresh water; glycerol and / or other alcohols such as ethanol, propanol or isopropanol; higher alcohols such as isoamyl or propylene glycol; sodium chloride and / or other salts. Composition according to any one of Claims 1 to 11, characterized in that it is for use as a medicine. Composition according to claim 18, characterized in that at least one bioactive compound is comprised in concentrations in the range of 1 to 8,000 ppm, preferably from 1 to 1,500 ppm. 5/6 Composition according to claim 18 or 19, characterized in that the acceptable vehicle comprises one or more vehicles selected from DMSO, lactose, stearate, starch, carbonates and / or, bicarbonates, methylcellulose, sodium alginate, ethanol, glycerol, sorbitol , sterile distilled water, sterile saline, osmolarity buffers, pH and / or phosphate; zinc oxide and / or cellulose gum. 21. Method for the prevention, mitigation and control of biogenic processes, characterized by understanding the application of the composition as defined in any one of claims 1 to 14 in contact times suitable for the effective reduction of the contaminating population of bacteria, in which the bacteria are planktonic or sessile and with isolated or combined action. 22. Method according to claim 21, characterized in that the contact time is about 4 to about 24 hours for planktonic bacteria and about 15 days for sessile bacteria. 23. Method according to claim 21 or 22, characterized in that the bacteria are BRS originating from different sites in the oil industry, both surface and subsurface, from oil and gas process and storage vessels, from fluid collected in oil production system subject to acidification (souring) and other inshore and offshore oil installations, and / or Total Heterotrophic Anaerobic Bacteria (BANHT) consortium to BRS. 24. Method for inhibiting the growth of bacteria, characterized in that it comprises the application of the composition as defined in any one of claims 1 to 11 and 15a17 in contact times suitable for the effective reduction of the bacterial population of the contaminated environment to be treated. 25. Method according to claim 24, characterized in that the contact time is 18 to 24 hours. 26. The method of claim 24 or 25, characterized in that the bacteria are strains including some found in oil reservoirs located in deep sea waters. 27. The method of any one of claims 24 to 26, characterized in that the bacteria are of the genus Bacillus, preferably Bacillus licheniformis, Bacillus amyloliquefasciens, Bacillus subtilis, Bacillus cereus, or Paenibacillus macerans. 6/6 28. Use of the composition as defined in any one of claims 1 to 11 and 18 to 20, characterized in that it is for the manufacture of a medicament for inhibiting bacterial targets. 29. Use according to claim 28, characterized in that the bacterial targets are S. aureus, E. coli, E. faecalis, P. aeruginosa, S. Enterítidis, S. Typhi, S. flexneri, or Paenibacillus macerans or species of the genus Bacillus, preferably Bacillus licheniformis, Bacillus amyloliquefasciens, Bacillus subtilis or Bacillus cereus. 30. Use of the composition as defined in any one of claims 1 to 14, characterized in that it is for the prevention, mitigation and control of biogenic processes, preferably souring, corrosion influenced by microorganisms and buffering of oil wells, or inhibiting the degradation of corrosion inhibitors in diesel transport pipelines. 31. Use of the composition as defined in any one of claims 1 to 11 and 15 to 17, characterized in that it is in asepsis, disinfection and sterilization of domestic, hospital and industrial environments, including the prevention of contamination and reduction of infection in a hospital environment.