ES2677155B1 - PORTA-COUPONS FOR MICROBIAL BIOPELICULES, SYSTEM AND REACTOR OF BIOPELICULAS THAT INCLUDES THE COUPON-HOLDER, AND METHOD TO OPERATE THE REACTOR - Google Patents

Description

DESCRIPTION

PORTA-COUPONS FOR BIOPEL ^ ULAS MICROBIAL, SYSTEM AND REACTOR OF BIOPELFCLES THAT INCLUDES THE COUPON-HOLDER, AND METHOD TO OPERATE THE

REACTOR

Sector of the technique

The present invention relates in general, in a first aspect, to a coupon holder for microbial biopellicles, and more particularly to a coupon holder that includes an electrical transducer and / or an electrical sensor next to a target surface suitable for the growth of microbial biopellicles, and that enables the supply of electricity to the electric transducer and / or electrical sensor.

A second aspect of the present invention concerns a system that includes the cup holder of the first aspect.

A third aspect of the present invention concerns a biopelling reactor comprising the coupon holder of the first aspect.

A fourth aspect of the present invention relates to a method for operating a biopellicle reactor according to the third aspect of the invention.

State of the prior art

Biomedical devices are essential to prevent and treat a large number of diseases, however, its use is not without complications and one of the most serious is the infection associated with its use. This type of infection is related to the ability of microorganisms to colonize the surface of a biomaterial and form in it a complex microbial community called "biofilm", biocapa or biopellula. At present, there are a large number of laboratories and research centers working on the development of new technologies that prevent the formation of the microbial biopellicle on the surface of different biomaterials.

The CDC biopelling reactor is a device increasingly used today, given its obvious advantages: it has a methodology standardized by the "American Society for Testing Materials", it allows to integrate in each test multiple variables such as microorganism, time, biomaterial, drug , etc., and has been validated by the "Center for Disease Control of the United States" (CDC) as a method of choice to study the interaction between microorganisms and materials. However, the CDC biopelling reactor It has certain limitations when carrying out some studies, in particular associated with the types of coupon carriers available.

In this regard, in the state of the art, coupon carriers for microbial biopellins are known which combine the characteristics of the preamble of claim 1, that is to say they comprise an elongated body with a proximal end portion, a grip, and a distal end portion. manufactured from a biocompatible material and including at least one target surface suitable for the growth of microbial biopellicles.

All the coupon holders for microbial biopellins known in the state of the art, both those of the CDC reactors and those of other systems, are clearly improvable, since they are mere passive support elements of the coupons, membranes or equivalent elements that include to the target surface.

It appears, therefore, necessary to offer an alternative to the state of the art that covers the gaps found in it, through the provision of a coupon holder, and associated aspects, which considerably improves the known ones, in particular as regards to the benefits to be offered, which go beyond those relating to being mere passive supports.

Explanation of the invention

For this purpose, the present invention concerns, in a first aspect, a coupon holder for microbial biopellins, comprising, in a manner known per se, an elongated body with a proximal end portion, grip, and a distal end portion manufactured of a biocompatible material and including at least one target surface suitable for the growth of microbial biopellins.

Unlike the coupon carriers known in the state of the art, the one proposed by the first aspect of the present invention, characteristically, comprises an electric transducer and / or an electrical sensor arranged in said distal end portion, and means of electricity supply operatively connected with the electric transducer and / or the electric sensor.

Although, according to an example of embodiment, said target surface is part of a disc-shaped element known in the state of the art as a coupon, which is coupled to a housing defined in the distal end portion of the body of the holder -cupons, for other examples of embodiment such target surface is constituted by another class of element, such as a membrane, which is mounted directly on a region of the extreme distal portion of the body of the coupon holder or which is attached to it . In any case, In this document, the term "coupon holder" is understood as a carrier element of at least one of said target surfaces, independently whether it is through a coupon, a membrane or another equivalent element and suitable for the same purpose.

According to a preferred embodiment, the coupon holder of the first aspect of the invention comprises said electric transducer, which is an electroacoustic transducer, such as an interdigital transducer that includes a substrate of piezoelectric material (for example Lithium Niobate). ) and electrodes deposited thereon, configured and arranged to emit surface acoustic waves (SAW) on said target surface for the prevention of the formation of microbial biopellins thereon or the eradication thereof.

For an example of embodiment, the outer surface of said piezoelectric substrate defines the target surface.

However, for other embodiments, the target surface is not made of piezoelectric material, but is defined by the outer face of the distal end portion of the elongate body, or by coupons, membranes or equivalent elements attached, embedded or mounted on such a distal extreme portion, proximate to the electroacoustic transducer, in contact or without contact therewith, above, below or to one side thereof, according to a disposition that allows the generated acoustic waves to propagate through the regions that are not piezo-electric and that define the target surface.

In general, the electro-acoustic transducer is configured and arranged to emit surface acoustic waves with a frequency value ranging from thousands of kHz to tens of GHz, preferably greater than 10 MHz, when receiving from the media of electricity supply at least said determined electrical signal.

According to a preferred embodiment, said frequency value is substantially within a range between 33 and 37 MHz.

Advantageously, the electro-acoustic transducer is configured and arranged to emit surface acoustic waves with a power such that a power density of between 4 and 6 mW / cm2 is obtained on the target surface.

For another example of embodiment, the coupon holder of the first aspect of the present invention comprises said electric sensor (additionally to the transducer or alternatively), which is an electro-optical or electro-chemical sensor, configured and arranged to detect microbial biopellins formed on the target surface.

According to another embodiment example, the coupon holder comprises said electric transducer or an additional electric transducer, which is an electrothermal transducer (such as a heating resistor) configured and arranged to emit heat on said target surface. That is to say that, for the present example of realization, the plug holder comprises, additionally or in a complementary manner to the inclusion of said electro-acoustic transducer, an electro-thermal transducer.

According to an embodiment, said electrical supply means comprise electric conductors running through the elongate body from its proximal end portion to its distal end portion, with first ends connected to electrical terminals of the electrical sensor and / or the electric transducer and a few second ends located in the extreme proximal portion.

Advantageously, said electric conductors run through the interior of the elongated body, in its entirety or over a large part of its length (at least 80%), duly isolated both electrically and against electromagnetic interference, in general, as they are constituted by respective coaxial cables. Alternatively, the electrical conductors run on the outside of the elongate body, also properly insulated and protected.

For a realization example, the coupon holder of the first aspect of the present invention comprises two connection terminals electrically connected to the second ends of the electric conductors, arranged in the proximal end portion of the elongated body and suitable for electrical connection with a Remote electric power generator to receive at least one electrical signal from it.

For another example of embodiment, the coupon holder of the first aspect of the present invention comprises a local electrical power generator arranged in the proximal end portion of the elongate body, and comprising respective output terminals connected to the second ends of the conductors. electrical, the local electrical power generator being configured to generate at least one electrical signal and provide it by their respective output terminals.

Alternatively, the aforementioned electricity supply means comprise a local electrical / electronic unit and a remote electrical / electronic unit communicated with one another, via cable or wirelessly, cooperating to generate at least one electrical signal and providing it via output terminals of the local electrical / electronic unit to electrical terminals of the electric sensor and / or the electric transducer connected to said output terminals.

Preferably, the coupon carrier of the first aspect of the present invention is configured and dimensioned for mounting, removably, in a head of a biopellicle reactor, by introducing it in a through opening thereof so that the The extreme end portion and the proximal end portion of said elongate body remain, respectively, inside and outside a biopellicle reactor vessel on which the head is positioned.

Even more preferably, said biopellicle reactor is of the CDC type. Different models of such a reactor are defined and marketed by the company "BioSurface Technologies Corporation", such as model CBR-2203. The definition of such models made by such company is incorporated herein by reference, for, among other purposes , defining the dimensions and dimensional relationships of all the elements of the reactor and, in particular, of the coupon holder of the present invention, whose dimensions are suitable for use in a CDC reactor.

A second aspect of the present invention concerns a system comprising the coupon holder of the first aspect according to the realization example for which it comprises two separate connection terminals, and to said remote electric energy generator, which in turn comprises respective output terminals connected to said connection terminals, the remote electric power generator being configured to generate at least one electrical signal and provide it by their respective output terminals.

In a third aspect, the present invention concerns a biopellicle reactor, comprising, in a manner known per se:

- a vessel for containing a culture medium in continuous agitation;

- coupon holders for microbial biopellins, each of which comprises an elongate body with a proximal end portion, grip, and a distal end portion made of a biocompatible material and including at least one target surface suitable for growth of microbial biopellins; Y

- a support head coupled to the mouth of said vessel, in the manner of a lid, and comprising through openings for the passage and mounting, removably, of said coupon holders.

Characteristically, in the biopellier reactor proposed by the third aspect of the present invention, at least one of said coupon carriers is a coupon carrier defined according to the first aspect of the invention.

According to a preferred embodiment, the biopellicle reactor of the third aspect of the present invention is of the CDC type.

The present invention also concerns, in a fourth aspect, a method for operating a biopellicle reactor according to the second aspect, comprising:

- introducing a culture medium and a microbial inoculum into the biopellicle reactor vessel;

- Attach the support head to the mouth of the vessel, closing it,

- mounting at least one coupon holder for microbial biopellins, defined according to the first aspect of the present invention, in the support head, previously or after coupling the support head in the mouth of the vessel;

- supplying at least one electrical signal to the electric transducer and / or electrical sensor arranged in the distal end portion of the elongate body of the coupon holder;

- incubating the microbial inoculum for a certain time, subjected to continuous agitation within the culture medium, and maintaining said supply of said electrical signal to the electric transducer and / or electrical sensor; and

- inspect the target surface, at least after said determined time, in order to detect possible microbial biopellins on it.

For a preferred embodiment, the method of the fourth aspect of the present invention comprises using a coupon holder with an electro-acoustic transducer, and said step of supplying at least one electrical signal comprises supplying the electro-acoustic transducer for at least one electrical signal determined to cause the transducer to emit surface acoustic waves on the target surface for the prevention of the formation of microbial biopellins thereon or the eradication thereof.

In general, the aforementioned electrical signal is an alternating signal configured to cause the electro-acoustic transducer to emit surface acoustic waves within a range from thousands of kHz to tens of GHz, preferably greater than 10 MHz.

According to a preferred embodiment, the alternating signal is configured to cause the electro-acoustic transducer to emit surface acoustic waves with a frequency value substantially within a range of between 33 and 37 MHz.

Advantageously, the determined electrical signal is an alternating signal configured to cause the electro-acoustic transducer to emit surface acoustic waves with a power such that a power density of between 4 and 6 mW / cm2 is obtained on the target surface, preferably substantially 4. , 9 mW / cm2.

Other frequency ranges, different from those previously indicated as preferred, as! like other values of power density, they are also covered by the present invention (in its four aspects), for other realization examples, adapting these to the type of transducer used. For example, for the case that this is an interdigital transducer, the frequency range will depend on the material of the piezoelectric substrate, its geometry and dimensions and the geometry and disposition of the electrodes on it. The greater the number of fingers of the comb-shaped electrodes, the narrower the frequency range, which will require a lower power density to generate the same prevention / eradication effect of microbial biopellicles. The upper limit of the range of working frequencies is given by the ability to place electrodes on the piezoelectric substrate with sufficiently small separation between the fingers.

In any case, according to the present invention, the power density will be sufficient to prevent / eradicate the formation of microbial biopellins.

According to an example of embodiment, said microbial inoculum includes at least one clinical isolate of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, or a combination thereof.

According to an example of embodiment, the reaction volume, culture medium plus microbial inoculum, is such that it does not cover the target surface, generating a humid chamber environment contaminated inside the biopellicle reactor vessel, although the method of fourth aspect of the present invention is also valid for the case that the target surface is immersed in the reaction volume.

With regard to the aforementioned determined time, this is within a range of several hours to several days, being the necessary for the formation of biopellicles on surfaces that were not subject to the effects of the transducer and / or electrical sensor, and it will be a function of the type of bacteria and the conditions used.

Brief description of the drawings

The foregoing and other advantages and features will be more fully understood from the following detailed description of some realization examples with reference to the accompanying drawings, which must be taken by way of illustration and not limitation, in which:

Figures 1a and 1b are two perspective views, taken from below and from above, respectively, of the coupon holder of the first aspect of the present invention, for an example of embodiment;

Figure 2 shows, schematically, an embodiment of the coupon holder and the biofilm reactor proposed by the present invention, where the electrical conductors have been drawn by discontinuous lines;

Figure 3 shows, schematically, the biofilm reactor proposed by the present invention, for an example of embodiment for which includes two coupon carriers defined according to the first aspect of the present invention, only the transducer of one of the which has been supplied with electricity, during an experimental trial;

Figures 4, 5 and 6 are graphs showing the results obtained by the aforementioned experimental test, in terms of quantification of viable cells of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, respectively.

Figures 7a to 7d are graphs showing the conversion of electrical energy in surface acoustic waves as a function of the frequency applied to the transducer of the cup holder of the first aspect of the present invention, for an example of embodiment, during a process of assembly, sterilization , operation and finalization of the experimental trial.

Detailed description of some examples of realization

In Figures 1a, 1b and 2, the coupon holder P for microbial biofilms is illustrated, for exemplary embodiments, for which the coupon holder P comprises an elongate body 1 with a proximal proximal portion 1a, grip, and a distal end portion 1b made of a biocompatible material and including at least one target surface D suitable for the growth of microbial biofilms, as well as an electroacoustic transducer Z, configured and arranged to emit surface acoustic waves on the target surface D for prevention of the formation of microbial biofilms on the same or the eradication thereof, and electricity supply means comprising electrical conductors W running inside the elongate body 1 from the proximal end portion 1a to the distal end portion 1b, with a few first ends connected to electrical terminals of the electric transducer Z (connection not shown) and second ends located in the proximal end portion 1a.

In Figures 1a, 1b and 2, the target surface D has been indicated as defined by the transducer Z itself. This is because in this case, the transducer Z is an interdigital transducer comprising metal electrodes E in the form of a comb. , interleaved, deposited on a piezoelectric substrate L, whose outer surface defines said target surface D.

For the embodiment illustrated in Figures 1a, 1b and 2, the transducer Z is mounted (by the screws T for the embodiment of Figure 2) on an insulating support plate S, for example of Nylon.

The present inventors have manufactured a prototype of the coupon holder P of the first aspect of the present invention according to the design of Figures 1a and 1b, where the elongate body 1 has been manufactured in Nylon (although various materials can be used, such as for example PET or stainless steel) and having the same external dimensions as a standard coupon holder for a CDC biopeaker reactor, in particular according to the CBR-2203 model of "BioSurface Technologies Corporation".

As shown in Figures 1a and 1b (and more schematically, in Figure 2), at the top, the coupon holder P has two terminals b of the SMA or BNC type that allow electrical connection with a external equipment The connection terminals b are electrically connected to the second ends of the electric conductors W and are suitable for electrical connection with a remote electric power generator (not shown) to receive the signal or electrical signals from it. The electric conductors W are generally constituted by two coaxial cables that allow the transmission of alternating electrical signals of high frequency with low attenuation and reduced noise, and run the central axis of the coupon holder P carrying the electric current from the upper terminals b to some lower terminals r.

As indicated above, in the distal end portion 1b of the elongate body 1, an insulating support plate S, for example of Nylon, is coupled to the lower terminals r of the coupon holder P by means of four manufactured metal contacts p, for example, in gold. On this insulating support plate S the transducer or transducers Z, in general of the interdigital type, is mounted (for example by means of plastic screws T, see Figure 2), electrically connecting (connection not shown) to the lower terminals r, in general through at least part of the metal contacts p. The electric current supplied by the remote generator accedes to the surface of the transducer or Z transducers from the lower terminals r, passing through the contacts p and of various metal filaments (not shown), for example aluminum, gold, silver or copper.

The prototype coupon holder P manufactured by the present inventors can be used in any standard CDC biopeaker reactor unit, and allows to bring electricity to the biomaterial under study. The presented design is adapted to concretely study the effect of surface acoustic waves (SAWs) generated on the surface of a piezoelectric material, ie the piezoelectric substrate L of the transducer Z, and its possible benefits in the eradication of the microbial biopellicle.

In its design and fabrication the following fundamental ideas have been taken into account:

2) It must withstand sterilization by autoclaving at 121 ° C, 20 minutes.

3) It must have the same dimensions as the standard coupon holder in such a way that the new modified coupon holder can be used in any CDC-type biopelling reactor unit.

4) It must be able to transmit an electrical impulse from an external generator to the surface of the biomaterial under study.

In Figure 2 is schematically illustrated also the CDC biopellier reactor that includes the coupon holder P, according to a realization example, for which the reactor R comprises:

- a vessel V for containing a culture medium in continuous agitation;

- a support head H coupled to the mouth of the cup V, in the manner of a lid, and comprising through openings Ap for the passage and assembly, removably, of holders of coupons P; Y

- a coupon holder P for microbial biopellins, defined as explained above, and mounted, removably, on the head H, by introducing it in one of the through openings Ap thereof so that the distal end portion 1b and the proximal end portion 1a of the elongated body 1 remain, respectively, inside and outside the vessel V of the biopellier reactor R on which the head H. is placed.

Experimental test:

The present inventors carried out an experimental test with the coupon holder P and reactor R of the present invention, also applying the method of the present invention, as described below:

For the test, a CDC biopelling reactor unit was equipped with two standard coupon carriers, loaded with titanium and teflon control wafers (not shown) that define respective target surfaces for the growth of biopellicles, and two carrier-defined coupons according to the first aspect of the present invention loaded with wafers A, B of Lithium Niobate forming respective piezoelectric substrates L (of respective Z transducers) defining respective target surfaces D. Surface acoustic wave generation tests were performed before sterilization and after sterilization, with positive results. The electro-acoustic transducer Z of one of the lithium niobate wafers, in particular of the one identified in Figure 3 as B, remained disconnected throughout the experiment, as a control, while the Z transducer of the rest of the wafers, in particular of the wafer A in Figure 3, it was kept electrically powered throughout the entire test.

The test was performed using clinical isolates of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, which were previously characterized and classified as strongly biopellula forming, according to the crystal violet staining in 96-well plates.

50ml of culture medium was added to the CDC Biofilm Reactor and 1ml of the microbial inoculum that contributed a total of 108 cfu / ml. As illustrated schematically in Figure 3, in this situation, the level Lv of the reaction volume V was such that it did not cover the wafers A, B, generating a contaminated wet chamber environment.

The frequency of the Z-transducer (not shown in Figure 3) of the Wafer A was adjusted in a range between 33 and 37 MHz and an amplitude of 20 dBm and was maintained in this situation for 24 hours, maintaining the reaction volume in agitation Continue using the agitator S.

After 24 hours of incubation in reactor R of CDC biopellins, wafers A and B were removed and subjected to a vortexing procedure at 2500 rpm and sonication at 50 kHz to release the biopellicle. The number of viable cells adhered to the surface of wafers A, B and control biomaterials was determined by a quantified culture of the broth after sonication on BHI agar. This culture-after-sonication procedure is a standard protocol in clinical microbiology for the study of infections related to biomedical devices such as prostheses, catheters, etc. This protocol allows to free the cells adhered to a surface and collect them in a liquid medium for later, by quantitative microbiological culture, proceed to determine the number of them that was present on the surface.

In Figures 4, 5 and 6, in conjunction with tables 1, 2 and 3 associated therewith, which are included below, the viable cell count of C. albicans, P. aeruginosa and S. aureus can be observed. respectively, present on the surface of the control biomaterials (titanium and PTFE), of the connected transducer (Wafer A) and of the disconnected transducer (Wafer B).

Table 1 (relative to Figure 4):

Biomaterial Count (ufc / cm2) Count (log ufc / cm2)

TITANIUM 951 2.98

TEFLON 618 2.79

OBLEA TO 0 0.00

OBLEA B 1471 3.17

Table 2 (relative to Figure 5):

Biomaterial Count (ufc / cm2) Count (log ufc / cm2)

TITANIUM 2705882 6.43

TEFLON 2098039 6.32

OBLEA TO 0 0.00

OBLEA B 3019608 6.48

Table 3 (relating to Figure 6):

Biomaterial Count (ufc / cm2) Count (log ufc / cm2)

TITANIUM 313725 5.50

TEFLON 7255 3.86

OBLEA TO 0 0.00

OBLEA B 7059 3.85

Observing Tables 1, 2 and 3, and Figures 4, 5 and 6, it can be stated that the results obtained clearly demonstrate that the implementation of the present invention in the experimental test described here has been able to prevent the colonization of the surface of the A wafer with an efficiency of 100% using a frequency range between 33 and 35 MHz and an amplitude of 20 dbm. The Z-transducer has been effective in preventing colonization by three different microorganisms: a fungus (Candida albicans), a gram-positive bacterium (Staphylococcus aureus) and a gram-negative bacterium (Pseudomonas aeruginosa).

Finally, in the graphs of Figures 7a to 7d can be seen the conversion of electrical energy in surface acoustic waves as a function of the frequency applied to the transducer Z. The optimal operating was between 33 and 37 MHz, a value that did not suffer significant changes during the assembly process (Figure 7a), autoclave sterilization (Figure 7b), operation (Figure 7c) and completion of the experimental test at 24 hours (Figure 7d).

The experimental test was repeated for other times of incubation, in particular for 48 and 72 hours, offering in both cases also positive results (not illustrated).

A person skilled in the art could introduce changes and modifications in the described embodiment examples without departing from the scope of the invention as defined in the appended claims.

Claims (26)

1. - Coupon carrier (P) for microbial biopellins, comprising an elongated body (1) with a proximal end portion (1a), grip, and a distal end portion (1b) made of a biocompatible material and including the less a target surface (D) suitable for the growth of microbial biopellins, characterized in that it comprises an electric transducer (Z) and / or an electrical sensor disposed in said distal end portion (1b), and electricity supply means operatively connected to said electrical transducer (Z) and / or said electric sensor. 2. - coupon holder (P) according to claim 1, comprising said electric transducer (Z), which is an electro-acoustic transducer, configured and arranged to emit surface acoustic waves on said target surface (D) for prevention of the formation of microbial biopellins on the same or the eradication of them. 3. - Coupon carrier (P) according to claim 2, wherein said electro-acoustic transducer (Z) is configured and arranged to emit surface acoustic waves with a frequency value within a range of thousands of kHz up to the tens of GHz, when receiving from said means of electricity supply at least one determined electrical signal. 4. - coupon holder (P) according to claim 3, wherein said electro-acoustic transducer (Z) is configured and arranged to emit surface acoustic waves with a frequency value greater than 10 MHz, when receiving from the means of electricity supply at least said certain electrical signal. 5. - Coupon carrier (P) according to claim 4, wherein said electro-acoustic transducer (Z) is configured and arranged to emit surface acoustic waves with a frequency value substantially within a range of between 33 and 37 MHz , when receiving from said means of electricity supply at least one determined electrical signal. 6. - coupon holder (P) according to any one of claims 2 to 5, wherein said electro-acoustic transducer (Z) is configured and arranged to emit surface acoustic waves with a power such that a power density of between 4 and 6 mW / cm2 on the target surface (D). 7. - Coupon carrier (P) according to any one of claims 2 to 6, wherein the electro-acoustic transducer (Z) is an interdigital transducer comprising metal electrodes (E) in the form of a comb, interlaced, deposited on a piezo-electric substrate (L) whose outer surface defines said target surface (D). 8. - Coupon carrier (P) according to any one of the preceding claims, comprising said electrical sensor, which is an electro-optical or electro-chemical sensor, configured and arranged to detect microbial biopellins formed on the target surface (D) ). 9. - coupon holder (P) according to any one of the preceding claims, comprising said electric transducer (Z) or an additional electric transducer, which is an electro-thermal transducer configured and arranged to emit heat on said target surface ( D). 10. - coupon holder (P) according to any one of the preceding claims, wherein said electricity supply means comprise electrical conductors (W) that run through said elongate body (1) from said proximal end portion (1a) to said distal end portion (1b), with first ends connected to electrical terminals of said electrical sensor and / or said electrical transducer (Z) and second ends located in the proximal end portion (1a). 11. - coupon holder (P) according to claim 10, comprising two separate terminals (b) electrically connected to said second ends of said electrical conductors (W), arranged in the proximal end portion (1a) of the elongated body ( 1) and suitable for electrical connection with a remote electric power generator to receive at least one electrical signal from it. 12. - Coupon carrier (P) according to claim 10, comprising a local electrical power generator arranged in the proximal end portion (1a) of the elongated body (1), and comprising respective output terminals connected to said second ends of said electrical conductors (W), said local electrical power generator being configured to generate at least one electrical signal and provide it by said respective output terminals. 13. - coupon holder (P) according to any one of the preceding claims, configured and sized for mounting, removably, in a head (H) of a biopellicle reactor (R), by introducing it in a through opening (Ap) thereof so that the distal end portion (1b) and the proximal end portion (1a) of said elongated body (1) remain, respectively, inside and outside a vessel (V) of the biopellicle reactor (R) on which the head (H) is placed. 14. - Coupon carrier (P) according to claim 13, wherein said biopellicle reactor (R) is of the CDC type. 15. - System, comprising the coupon holder (P) according to claim 11 and said remote electric power generator, which in turn comprises respective output terminals connected to said connection terminals (b), said generator being of remote electrical power configured to generate at least one electrical signal and provide it by said respective output terminals. 16. - Biopelicula reactor (R), comprising: - a vessel (V) for containing a culture medium in continuous agitation; - coupon holders (P) for microbial biofilms, where each of them comprises an elongated body (1) with a proximal end portion (1a), grip, and a distal end portion (1b) made of a biocompatible material and which includes at least one target surface (D) suitable for the growth of microbial biofilms; Y - a support head (H) attachable to the mouth of said vessel (V), in the manner of a lid, and comprising through openings (Ap) for the passage and mounting, removably, of said coupon holders (P) ); characterized in that at least one of said coupon carriers (P) is a coupon carrier defined according to any one of claims 1 to 14. 17. Biofilm reactor (R) according to claim 16, wherein the reactor (R) is of the CDC type. 18. - Method for operating a biofilm reactor (R) according to claim 16 or 17, comprising: - introducing a culture medium and a microbial inoculum into the vessel (V) of the biofilm reactor (R); - attach the support head (H) to the mouth of the vessel (V), closing it, - mounting at least one coupon holder (P) for microbial biofilms defined according to any one of claims 1 to 14 in the support head (H), previously or after coupling the support head (H) in the mouth of the vessel ( V); - supplying at least one electrical signal to the electric transducer (Z) and / or electrical sensor arranged in the distal end portion (1b) of the elongated body (1) of the coupon holder (P); - incubating the microbial inoculum for a certain time, subjected to continuous agitation in the culture medium, and maintaining said supply of said at least one electrical signal to the electric transducer (Z) and / or electrical sensor; and - inspect the target surface (D), at least after said determined time, in order to detect possible microbial biofilms on it. 19. - Method according to claim 18, comprising using a coupon holder (P) with an electro-acoustic transducer (Z), and wherein said step of supplying at least one electrical signal comprises supplying said electrode transducer. acoustic (Z) at least one electrical signal determined to cause the electro-acoustic transducer (Z) to emit surface acoustic waves on the target surface (D) for the prevention of the formation of microbial biofilms on it or the eradication thereof . 20. - Method according to claim 19, wherein said determined electrical signal is an alternating signal configured to cause the electro-acoustic transducer (Z) to emit surface acoustic waves with a frequency value within a range ranging from thousands from kHz to tens of GHz. 21. - Method according to claim 20, wherein said alternating signal is configured to cause the electro-acoustic transducer (Z) to emit surface acoustic waves with a frequency value greater than 10 MHz. 22. - Method according to claim 21, wherein said alternating signal is configured to cause the electro-acoustic transducer (Z) to emit surface acoustic waves with a frequency value substantially within a range between 33 and 37 MHz. 23. - Method according to claim 22, wherein the determined electrical signal is an alternating signal configured to cause the electro-acoustic transducer (Z) to emit surface acoustic waves with a power such that a power density of between 4 is obtained. and 6 mW / cm2 on the target surface (D). 24. Method according to any one of claims 18 to 23, wherein said microbial inoculum includes at least one classical isolate of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, or a combination thereof. 25. Method according to any one of claims 18 to 24, in which the volume of reaction, culture medium plus microbial inoculum, is such that it does not cover the target surface (D), generating a contaminated wet chamber environment inside of the vessel (V) of the biofilm reactor (R). 26. - Method according to any one of claims 18 to 25, in said determined time is within a range ranging from several hours to several days.