Definitions

• the present invention relates to a hygiene detection device which can be used in the fields such as food, water, agriculture, medicine, biology and chemistry, and that enables determination of toxic characteristics caused by the contamination of bacteria that has the ability to form indole by decomposing tryptophan amino acid.
• the methods allowing detection under 24 hours are quick-detection methods. There are quick detection devices and quick detection sets for allowing this. The detection times have been lowered by these quick detection sets, however for the bacteria enrichment process before the detection, 18-24 hours are needed.
• Table 2 and Table 3 discloses the total number of samples arriving at the 99 private food control laboratories as of 2017, according to the annual report of 2017 of the Directorate General of Control of Ministry of Food and Livestock of
• the price list of the Ankara Food Control Laboratory Administration as of 2017 for the analysis of Coliform Bacteria / Number of fecal bacteria in food and feed for 1 kg is 171 TL and analysis of Coliform Bacteria / Number of fecal bacteria in water for 1 It is 155 TL.
• the present invention is developed by the inspiration of the state of the art, and it aims to solve the above-mentioned problems.
• the objective of the invention is to determine the toxic characteristics caused by the contamination of bacteria that has the ability to form indole by decomposing tryptophan amino acid, which can be used in the fields such as food, water, agriculture, medicine, biology and chemistry.
• Another objective of the invention is to provide insight about the consumability of foods. Yet another objective of the invention is to reveal the contamination of ready-to- eat foods.
• Yet another objective of the invention is to detect the products having low hygienic qualities by search tests.
• Yet another object of the invention is to reduce the workload of the food control laboratories.
• Yet another object of the invention is to provide added value as per community health. Yet another objective of the invention is to provide a solution that is quick, economic and portable.
• FIG. 1 is a schematic illustration of the inventive hygiene detection device. Reference numerals 1.
• Hygiene detection device
• the present invention relates to a hygiene detection device (1) which can be used in the fields such as food, water, agriculture, medicine, biology and chemistry, and that enables determination toxic characteristics caused by the contamination of bacteria that has the ability to form indole by decomposing tryptophan amino acid.
• the proposed hygiene detection device (1) enables detection of toxic characteristics caused by the contamination of bacteria that has the ability to form indole by decomposing tryptophan amino acid.
• E. coli and V. cholera can secrete indole up to 0.6 mM in rich media.
• discrimination between the types of bacteria (Salmonella (-), Edwardsiella (+), E. coli(+), Klebsiella (-), Enterobacter (+), and species (P. multocidea (+), P. haemolytica (-), P. mirabilis (-), P. vulgaris (+)) could easily be done by the inventive hygiene detection device (1).
• the inventive hygiene detection device (1) provides insight about the consumability of foods, reveals the present contamination of ready-to-consume foods, and detects the products having low hygienic qualities by search tests. By the virtue of the hygiene detection device (1) the workload of the food control laboratories is reduced and a quick, economic and portable solution which provides an added value in terms of the community health is presented.
• the hygiene detection device (1) Since the hygiene detection device (1) is used for detection of hygiene of food and water, it prevents the spreadable diseases and the food-related deaths. Renal failures, diarrhea, vomiting and the economic turmoil and social traumas caused by insufficient hygiene are reduced by means of the hygiene detection device (1). By the use of the hygiene detection device (1), the community health is improved and the length of the treatment process of the patients and the economic expenses during the treatment are reduced.
• the hygiene detection device (1) could be used for determining the contamination level of the waters and is therefore efficient in management of water resources. For this reason, the commercialization potential of the hygiene detection device (1) in the fields of food, water, agriculture and especially in health sector, and at the same time, it has a wide field of application.
• the hygiene detection device (1) provides a study about the bacteria that has the ability to form indole.
• Indole is a nucleus detecting molecule which mediates the intercellular signals in bacteria.
• Indole has various important applications in pharmaceutical industry and is a bioindicator in biological and clinical samples. Specific analysis for detecting the indole amount are compicated and they require expensive equipment and high-level education. For this reason, Kovak’s test, which non-specifically detects indole analogues in samples, is used. For example, in the presence of 5 mM of tryptophan, E. coli bacteria could produce about 3.3 mM of indole.
• the indole controls various aspects of bacterial physiology in indole-producing bacteria such as spore formation, plasmid stability, drug resistance, biofilm generation and virulence.
• Some of the Gram positive bacteria of nearly 85 indole-producing bacteria species are as follows: Bacillus alvei, Clostridium novyi, Clostridium limosum, Clostridium tetani, Corynebacterium acnes, Desulfitobacterium hafniense, Enterococcus faecalis, Symbiobacterium thermophilum.
• Some of the Gram-negative bacteria are as follows: Aeromonas hydrophila, Aeromonas sivilar, Bacteroides sp., Chromobacterium violaceum, Citrobacter freundii, Citrobacter koseri, Edwardsiella tarda, Enterobacter aerogenes, Escherichia albertii, Escherichia coli, Escherichia fergusonii, Flavobacteria, Fusobacterium polymorphum, Haemophilus influenzae, Klebsiella ornithinolytica, Klebsiella planticola, Pantoea topaklan, Pasturella pneumotropica, Proteus vulgaris, Pseudovibrio sp., Shigella boydii, Shigella flexneri, Sphaerophorus varius, Vibrio cholerae, Yersinia intermedia, Yersinia enterocolitica, Yersinia frede
• the bacteria to be detected are the bacteria that has the ability to form indole using the tryptophan. These bacteria are indole positive bacteria. The most important ones of these bacteria, Edwadsiella, Klebsiella, Vibrio and especially Escherichia coli are indicators of fecal contamination. These bacteria have the ability to form indole by decomposing tryptophan, which is an amino acid. Indole is a test enabling to distinguish between the genus and species of bacteria, especially most coliform bacteria. Indole test is as follows: The inventive hygiene detection device (1) takes advantage of biological reactions during its utilization. The inventive hygiene detection device (1) has determined that the bacteria synthesize sufficient indole in 4-8 hours.
• the microorganisms are incubated at 37 °C for 24-58 hours after being passaged to a suitable broth (generally TB).
• a suitable broth generally TB.
• the reason for keeping it at 37 °C is that it provides a suitable temperature condition for both bacterial reproduction and for the bacteria to synthesize indole. As the bacteria in the sample reach a certain number at the end of incubation (4-8 hours), sufficient indole is produced.
• the hygiene detection device (1) has a major importance for testing the convenience of hygiene and sanitation conditions, during the harvest and transportation of food products in farms and ranches or during its processing in factories, during packaging, during its distribution at markets, restaurants and food serving places or homes.
• a quick and practical testing is provided based on the traceability of the hygiene of food, water and feed in this chain. Not being able to provide this safeness causes distrust and loss of customers, reduction in consumption and sales, loss of reputation and reduction in market share. More important from these economical losses is the harm done on the human beings by these non-hygienic conditions.
• the detection of the hygiene of food and water in short time will make it easier for the food products to be produced and served for consumption in suitable conditions and in conformity with the regulations.
• the hygiene detection device (1) could also be used in institutions such as medical institutions, food and water producing companies, hotels, cleaning and sanitation companies, restaurants, military units, recreational facilities, businesses selling raw meat, businesses producing ready- to-consume food, textile factories and such entities.
• the hygiene detection device (1) can detect fecal bacteria in analysis of drinking water and utility water and food, in a short time.
• the hygiene detection device (1) schematically illustrated in Figure 1 comprises;
• ⁇ at least one battery (2) which provides the electrical power required by the components in order for them to work
• ⁇ at least one test cartridge (3) which amplifies the bacteria level from 10 3 to 10 5 such that the indole reaction takes place and the red ring occurs
• ⁇ at least one resistor (4) for heating the tube having the food or water sample passaged with a broth (generally TB) having tryptophan amino acid, and obtaining optimal temperature, thereby allowing the bacterial reproduction and preserve aliveness
• thermometer setup (5) enabling the incubation continuously for 4-8 hours at the temperature, 37 °C, obtained by the said resistor (4), enabling the enzyme produced by the bacteria in this incubation to react with tryptophan amino acid, and thereby enabling the production of sufficient indole from the tryptophan amino acid by way of deamination,
• ⁇ at least one optical reader (6) for sensing the red color formed on the upper part of the tube upon the reaction of the indole produced by the bacteria with the Kovak's reactive, after adding on the tube a sufficient amount of the Kovak's reactive which is a biochemical consisting of isoamyl alcohol, paradimethylaminobenzaldehyde and hydrochloric acid, after the bacteria reaches a certain level after the incubation period and produces indole, if there are bacteria present in the food or water sample which are kept below a certain temperature,
• thermometer setup (5) which enables to hold the food or water sample at an optimum temperature by determining the operating manner of the said resistor (4) according to the said data, which analyses the frequency of the red color from the data it receives from the optical reader (6), which detects upon the said analysis that the said food or water sample contains a bacteria capable of forming indole or not, and which distinguishes the genus and species of a bacteria present in the said sample, and
• the number of bacteria is amplified from a level of 10 3 to a level of 10 8 .
• the number of bacteria is amplified from a level of 10 3 to a level of 10 5 . This amount is sufficient for the indole reaction to take place and for the red ring.
• TB is a broth containing tryptophan.
• the tryptophanase enzyme produced by E. coli or such bacteria reacts with the tryptophan, and pyruvic acid, ammonia and indole is formed from the tryptophan by deamination.
• an incubation environment of 37 C is provided for the bacteria, and the bacteria is left for incubation at 37 C for 4-8 hours.
• 0.5 ml of Kovak's reactive is dripped on the indole that is produced by a bacteria such as E. coli in a sufficient amount, that is, it contains sufficient indole for detecting the maroon layer formed on upper surface of the Eppendorf in which the reaction takes place.
• the Kovak's reactive is a biochemical reactive consisting of isoamyl alcohol, paradimethylaminobenzaldehyde and hydrochloric acid.
• the indole produced by E. coli and such bacteria reacts with the para-dimethylaminobenzaldehyde, and they form a maroon colored layer on the surface of the Eppendorf in which they are located.
• the optical reader (6) senses the frequency of this maroon color, and the presence of bacteria, if any, is shown to the user via the LCD screen (8).

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• 2019
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