• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
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    • 专利摘要:
    Porta-coupons for microbial biofilms, biofilm system and reactor that includes the coupon holder, and method to operate the reactor. The coupon holder (P) comprises an elongated body (1) with a proximal end portion (1a), gripper, 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, an electrical transducer (Z) and/or an electrical sensor arranged in the distal end portion (1b), and electricity supply means operatively connected to the electrical transducer (Z) and/or a electrical sensor. The system comprises the coupon holder (P) of the present invention and a remote electric power generator. The reactor (R) comprises the coupon holder (P) of the present invention. The method is adapted to operate a biofilm reactor (R) defined according to the present invention.
    公开号:ES2677155A1
    申请号:ES201631733
    申请日:2016-12-30
    公开日:2018-07-30
    发明作者:Mauricio MORAIS DE LIMA；Julián HEREDERO CALERO；Pascual FERNÁNDEZ GONZÁLEZ；Andrés CANTARERO SÁEZ；Paula RAMÍREZ GALLEYMORE；Marcelo Ernesto FERNÁNDEZ RIVERO；Mónica GORDÓN SAHUQUILLO；Paulo VENTURA SANTOS
    申请人:Fundacion Para La Investigacion Hospital Universitario Y Politecnico La Fe de la Comunidad Valenciana；Forschungsverbund Berlin FVB eV；Fundacion Para La Investigacion Hospital Univ Y Politecnico La Fe de la Comunidad Valenciana；Universitat de Valencia；
    IPC主号:
    专利说明:

    COUPON HOLDER FOR MICROBIAL BIOPELLS, SYSTEM AND REACTOR OFBIOPELLS INCLUDING THE COUPON HOLDER, AND METHOD FOR OPERATING THEREACTOR
    Technical sector
    The present invention generally concerns, in a first aspect, a coupon holder for biofilm microbial particles, and more particularly a coupon holder that includes an electrical transducer and / or an electrical sensor close to a suitable target surface. for the growth of microbial biofilms, and that enables the supply of electricity to the electrical 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 biofilm reactor comprising the coupon holder of the first aspect.
    A fourth aspect of the present invention relates to a method for operating a biofilm reactor according to the third aspect of the invention.
    Prior art
    Biomedical devices are essential to prevent and treat a large number of diseases, however, their use is not without complications and one of the most serious is the infection associated with its use. This type of infections is related to the ability of microorganisms to colonize the surface of a biomaterial and form in it a complex microbial community called "biofilm", biofilm or biofilm. Currently, there are a large number of laboratories and research centers working on the development of new technologies that prevent the formation of microbial biofilm on the surface of different biomaterials.
    The CDC biofilm reactor is an increasingly used device today, given its obvious advantages: it has a methodology standardized by the "American Society for Testing Materials", allows multiple variables such as microorganism, time, biomaterial, drug to be integrated into each test , 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 biofilm reactor
    It has certain limitations when carrying out some studies, particularly associated with the types of coupon holders available.
    In this sense, in the state of the art, coupon holders are known for microbial biofilms that meet the characteristics of the preamble of claim 1, that is to say they comprise an elongated body with a proximal, gripping end portion, and a distal end portion made of a biocompatible material and that include at least one target surface suitable for the growth of microbial biofilms.
    All the coupon holders for microbial biofilms known in the state of the art, both those of the CDC reactors and those of other systems, are clearly improvable, since they constitute mere passive support elements of the coupons, membranes or equivalent elements that include to the target surface.
    It appears, therefore, that it is necessary to offer an alternative to the state of the art that covers the gaps found therein, by providing a coupon holder, and associated aspects, which considerably improves the known ones, in particular as regards to the benefits to offer, that go beyond those relative to being mere passive supports.
    Explanation of the invention.
    To this end, the present invention concerns, in a first aspect, a coupon holder for biofilm microbial particles, which comprises, in a known manner, an elongated body with a proximal, gripping end portion, and a distal end portion made of a biocompatible material and that includes at least one target surface suitable for the growth of microbial biofilms.
    Unlike the coupon holders known in the state of the art, the one proposed by the first aspect of the present invention, typically comprises an electric transducer and / or an electric sensor disposed in said distal end portion, and means of electricity supply operatively connected to the electrical transducer and / or the electrical sensor.
    Although, according to an exemplary embodiment, said target surface is part of a disk-shaped element known in the prior art as a coupon, which is coupled to a housing defined in the distal end portion of the body of the holder - coupons, for other embodiments such target surface is constituted by another kind of element, such as a membrane, which is mounted directly on a region of the distal end portion of the body of the coupon holder or that is fixed thereto . In any case, in this document the term coupon holder should be understood as a carrier element of at least one of such target surfaces, regardless of whether it is through a coupon, a membrane or another equivalent and suitable element therefor. finish.
    According to a preferred embodiment, the coupon holder of the first aspect of the invention comprises said electrical 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 biofilms on it or the eradication thereof.
    For an exemplary embodiment, the outer surface of said piezoelectric substrate defines the target surface.
    However, for other embodiments, the target surface is not of piezoelectric material, but is defined by the outer face of the distal end portion of the elongated body, or by coupons, membranes or equivalent elements attached, embedded or mounted on such an end distal portion, close to the electroacoustic transducer, in contact or without contact with it, above, below or to one side thereof, according to an arrangement that allows the generated acoustic waves to propagate through the regions that are not piezo -electric and defining the target surface.
    In general, the electro-acoustic transducer is configured and arranged to emit surface acoustic waves with a frequency value within a range from thousands of kHz to tens of GHz, preferably greater than 10 MHz, when receiving from the media of supplying electricity 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 embodiment, the coupon holder of the first aspect of the present invention comprises said electrical sensor (additionally to the transducer or alternatively), which is an electro-optical or electro-chemical sensor, configured and arranged to detect microbial biofilms formed on the target surface.
    According to another embodiment, the coupon holder comprises said electrical transducer or an additional electrical transducer, which is an electrothermal transducer (such as a heating resistor) configured and arranged to emit heat on said target surface. That is, for the present embodiment, the coupon holder comprises, additionally or in a complementary manner to the inclusion of said electro-acoustic transducer, an electro-thermal transducer.
    According to an exemplary embodiment, said electricity supply means comprise electrical conductors that run through the elongated 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 proximal end portion.
    Advantageously, said electrical conductors run through the inside of the elongated body, in whole or for a large part of its length (at least 80%), duly insulated both electrically and against electromagnetic interference, in general, being constituted by means of respective coaxial cables. Alternatively, the electrical conductors run on the outside of the elongated body, also properly insulated and protected.
    For an exemplary embodiment, the coupon holder of the first aspect of the present invention comprises two connection terminals electrically connected with the second ends of the electrical 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 exemplary embodiment, the coupon holder of the first aspect of the present invention comprises a local electric power generator disposed in the proximal end portion of the elongated body, and comprising respective output terminals connected to the second ends of the conductors. electrical, the local electric power generator being configured to generate at least one electrical signal and provide it by their respective output terminals.
    Alternatively, the said electricity supply means comprise a local electrical / electronic unit and a remote electrical / electronic unit communicated with each other, via cable or wirelessly, cooperating to generate at least one electrical signal and provide it through output terminals of the local electrical / electronic unit to electrical terminals of the electrical sensor and / or of the electrical transducer connected to said output terminals.
    Preferably, the coupon holder of the first aspect of the present invention is configured and sized for mounting, removably, in a head of a biofilm reactor, by inserting it into a through opening thereof so that the distal end portion and the proximal end portion of said elongated body remain, respectively, inside and outside a vessel of the biofilm reactor on which the head is placed.
    Even more preferably, said biofilm reactor is of the CDC type. Different models of such a reactor are defined and marketed by the company "Bio $ urface Technologies Corporation", such as the CBR-2203 model. 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 reactor elements 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 exemplary embodiment for which it comprises two connection terminals, and said remote electric power 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 biofilm reactor, comprising, in a manner known per se:
    - a glass to contain a culture medium in continuous agitation;
    - coupon holders for microbial biofilms, where each one comprises an elongated body with a proximal end portion, of grip, and a distal end portion made of a biocompatible material and which includes at least one target surface suitable for biofilm growth microbial; Y
    - a support head attachable to the mouth of said vessel, as a cover, and comprising through openings for the passage and assembly, removably, of said coupon holders.
    Typically, in the biofilm reactor proposed by the third aspect of the present invention, at least one of said coupon holders is a coupon holder defined according to the first aspect of the invention.

    According to a preferred embodiment, the biofilm reactor of the third aspect of the present invention is of the COCo type.
    The present invention also concerns, in a fourth aspect, a method for operating a biofilm reactor according to the second aspect, which comprises:
    - introducing a culture medium and a microbial inoculum into the biofilm reactor vessel;
    - attach the support head to the mouth of the vessel, closing it,
    - mount at least one coupon holder for microbial biofilms. defined according to the first aspect of the present invention, in the support head, before or after coupling the support head in the mouth of the vessel;
    - supply at least one electrical signal to the electrical transducer and / or electrical sensor disposed in the distal end portion of the elongated 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 electrical transducer and / or electrical sensor; and
    - inspect the target surface, at least after said determined time, in order to detect possible microbial biofilms 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 supply stage of at least one electrical signal comprises supplying the electro-acoustic transducer by 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 biofilms on it or the eradication thereof.
    In general, the said determined 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, other than those indicated above as preferred, as well as other power density values, are also covered by the present invention (in all four aspects), for other embodiments, these being adapted to the type of transducer used . For example, in 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 arrangement of the electrodes on it. The greater the range of frequencies of the comb-shaped electrodes, the frequency range will be narrower, which will require a lower power density to generate the same prevention / eradication effect of microbial biofilms. The upper limit of the working frequency range is given by the ability to arrange electrodes with a 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 biofilms.
    According to an exemplary embodiment, said microbial inoculum includes at least one clinical isolate of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus,
    or a combination thereof.
    According to one embodiment, the reaction volume, culture medium plus microbial inoculum, is such that it does not cover the target surface, generating a contaminated humid chamber environment inside the biofilm reactor vessel, although the method of Fourth aspect of the present invention is also valid in the event 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 biofilms to form on surfaces that were not subject to the effects of the transducer and / or electrical sensor, and It will depend on 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 examples of embodiment with reference to the attached drawings, which should be taken by way of illustration and not limitation, in which:
    Figures 1 a and 1 b 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 exemplary embodiment;
    Figure 2 shows, in a schematic way, an embodiment example of the coupon holder and the biofilm reactor proposed by the present invention, where the electrical conductors have been drawn by broken lines;
    Figure 3 shows, schematically, the biofilm reactor proposed by the present invention, for an exemplary embodiment for which it includes two coupon holders defined according to the first aspect of the present invention, only to 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 said 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 into 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 exemplary embodiment, during an assembly, sterilization process , operation and completion of the experimental trial.
    Detailed description of some embodiments
    In Figures 1a, 1 and 2, the coupon holder P for microbial biofilms is illustrated, for some embodiments, for which the coupon holder P comprises an elongated body 1 with an end portion proximal 1 a, of grip, and a distal end portion 1 b made of a biocompatible material and which includes 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 the prevention of the formation of microbial biofilms thereon or the eradication thereof, and electricity supply means comprising electric conductors W that run inside the elongate body 1 from the proximal end portion1a to the distal end portion1b, with a few first
    ends connected to electrical terminals of the electrical transducer Z (connection not shown) and second ends located in the proximal end portion1a.
    In Figures 1 a, 1 b and 2, the target surface O has been indicated as defined by the transducer Z itself. This is so because in such a case, the transducer Z is an interdigital transducer comprising metal electrodes E in shape. comb, interlaced, deposited on a piezo-electric 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 means of the screws T for the embodiment of Figure 2) on an insulating support plate S, for example 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 1 a and 1 b, where the elongated body 1 has been manufactured in Nylon (although various materials can be used, as per eg PET or stainless steel) and which has the same external dimensions as a standard coupon holder for an eDe biofilm reactor, in particular according to the CBR-2203 model of "BioSurface Technologies Corporalion".
    As can be seen 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 the electrical connection with a external equipment The connection terminals b are electrically connected to the second ends of the electrical conductors W and are suitable for electrical connection with a remote electrical power generator (not shown) to receive the electrical signal or signals from it. The electrical conductors W in general are constituted by two coaxial cables that allow the transmission of alternating electrical signals of high frequency with low attenuation and reduced noise, and run along the central axis of the coupon holder P carrying the electric current from the upper terminals b to 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 four fabricated metal contacts p, For example, in gold. The transducer or transducers Z, generally of interdigital type, are electrically connected (to the lower terminals r), in general, on this insulating support plate S (for example by means of plastic screws T, see Figure 2). through at least part of the metal contacts p. The electric current supplied by the remote generator thus accesses the surface of the transducer or
    Z transducers from the lower terminals r, passing through the contacts p and several metallic filaments (not shown), for example aluminum, gold, silver or copper.
    The prototype of coupon holder P manufactured by the present inventors can be used in any standard CDC type biofilm reactor unit, and allows electricity to be taken to the biomaterial under study. The design presented is adapted to specifically study the effect of surface acoustic waves (SAWs) generated on the surface of a piezoelectric material, that is to say the piezoelectric substrate L of the Z transducer, and its possible benefits in the eradication of the microbial biopelicula.
    The following fundamental ideas have been taken into account in its design and manufacture:
    2) Must withstand sterilization by autoclave at 121 ° C, 20 minutes.
    3) It must have the same dimensions as the standard coupon holder so that the new modified coupon holder can be used in any CDC type biofilm reactor unit
    4) Must be able to transmit an electrical impulse from an external generator to the surface of the biomaterial under study.
    Figure 2 also schematically illustrates the CDC biofilm reactor that includes the coupon holder P, according to an example of embodiment, for which the reactor R comprises:
    - a vessel V to contain a culture medium in continuous agitation;
    - a support head H coupled to the mouth of the vessel V, as a cover, and comprising through openings Ap for the passage and assembly, removably, of coupon holders P; Y
    - a coupon holder P for microbial biofilms, defined as explained above, and removably mounted on the head H, by insertion into one of the through openings Ap thereof so that the distal end portion 1 b Y the proximal end portion 1 a of the elongated body 1 remain, respectively, in and out of the vessel V of the biofilm reactor R on which the head is placed
    H.

    Experimental trial:
    The present inventors conducted 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 biofilm reactor unit was equipped with two standard coupon holders, loaded with titanium and Teflon control wafers (not illustrated) that define respective target surfaces for biofilm growth, and two coupon holders defined according to the First aspect of the present invention loaded with wafers A, S of Lithium Niobate that form respective piezoelectric substrates L (of respective Z transducers) that define respective target surfaces D. Surface sound 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 the one identified in Figure 3 as S, remained disconnected throughout the experiment, as a control, while the Z transducer of the other wafers, in particular of wafer A in Figure 3, it remained electrically powered throughout the test.
    The trial was performed using clinical isolates of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, which were previously characterized and classified as strongly biofilm-forming, according to the staining of crystal violet in 96-well plates.
    50ml of culture medium was added to the Siofilm Reactor CDC and 1 ml of the microbial inoculum which contributed a total of 108 cfu / ml. As schematically illustrated in Figure 3, in this situation, the level Lv of the reaction volume V was such that it did not cover the wafers A, S, generating a contaminated humid chamber environment.
    The frequency of transducer Z (not illustrated in Figure 3) of Wafer A was adjusted in a range between 33 and 37 MHz and an amplitude of 20 dSm and was maintained in this situation for 24 hours, keeping the reaction volume under stirring continue using the stirrer S.
    After 24 hours of incubation in the R reactor of CDC biofilms, wafers A and S were removed and subjected to a vortex agitation procedure at 2500 rpm and sonication at 50 kHz to release the biofilm. The number of viable cells adhered to the surface of wafers A, S and the control biomaterials was determined by a quantified culture of the broth after sonication on SHI agar. This culture procedure after sonication 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 release the cells adhered to a surface and collect them in a liquid medium and then, by quantitative microbiological culture, proceed to determine the number of them that was present on the surface.
    5 In Figures 4, 5 and 6, in conjunction with tables 1, 2 and 3 associated thereto, which are included below, the count of viable cells of C. albicans can be observed,
    P. aeruginosa and S. aureus respectively, present on the surface of the control biomaterials (titanium and PTFE), the connected transducer (Wafer A) and the disconnected transducer (Wafer B).
    10 Table 1 (relative to Figure 41:
    Biomaterial Count (cfu / cm2)Count (Iog cfu / cm2)
    TITANIUM 9512.98
    TEFLON 6182.79
    OBLEA A °0.00
    OBLEA B 14713.17
    Table 2 (relative to Figure 51:Biomaterial Count (cfu / cm2) Count (Iog cfu / cm2)
    TITANIUM 27058826.43
    TEFLON 20980396, 32
    °
    OBLEA A 0.00
    OBLEA B 30196086.48
    Table 3 (relative to Figure 51:Biomaterial Count (cfu / cm2) Count (Iog cfu / cm2)
    TITANIUM 3137255.50
    TEFLON 72553.86
    OBLEA A or0.00
    OBLEA B 70593.85
    Observing tables 1, 2 and 3, and Figures 4, 5 and 6, it can be affirmed that the results obtained clearly demonstrate that the implementation of the present invention in the experimental test described here, has been able to prevent surface colonization of wafer A with 100% efficiency using a frequency range between 33 and 35
    5 MHz and an amplitude of 20 dbm. The Z transducer has been effective in preventingcolonization by three different microorganisms: a fungus (Candida albicans), agram positive bacteria (Staphylococcus aureus) and a gram negative (Pseudomonasaeruginous).
    Finally, in the graphs of Figures 7a to 7d the conversion of
    10 electrical energy in surface acoustic waves as a function of the frequency applied to the Z transducer. The operating optimum was between 33 and 37 MHz, a value that did not undergo significant changes during the assembly process (Figure 7a), autoclave sterilization (Figure 7b), operation (Figure 7c) and completion of the experimental trial at 24 hours (Figure 7d).
    15 The experimental test was repeated for other incubation times, 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 embodiments without departing from the scope of the invention as defined in the appended claims.
    权利要求:
    Claims (26)
    [1]
    1.-Coupon holder (P) for biofilm microbial particles, comprising an elongated body (1) with a proximal end portion (1a), of grip, and a distal end portion (1 b) made of a biocompatible material and which includes at least one target surface (D) suitable for the growth of microbial biofilms, characterized in that it comprises an electric transducer (Z) and / or an electric sensor disposed in said distal end portion (1b), and operatively connected electricity supply means with said electrical transducer (Z) and / or said electrical sensor.
    [2]
    2. Coupon holder (P) according to claim 1, comprising said electrical 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 biofilms on it Or the eradication thereof.
    [3]
    3.-Coupon holder (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 from thousands of kHz to tens of GHz, upon receiving from said electricity supply means at least one determined electrical signal.
    [4]
    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, upon receiving at least said determined electrical signal from the electricity supply means.
    [5]
    5.-Coupon holder (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, upon receiving from said electricity supply means at least one determined electrical signal.
    [6]
    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]
    7. Coupon holder (P) according to any one of claims 2 to 6, wherein the electro-acoustic transducer (Z) is an interdigital transducer comprising comb-shaped, interlocking, deposited metal electrodes (E) on a part-electric substrate (l) whose outer surface defines said target surface (D).
    [8]
    8.-Coupon holder (P) according to any one of the preceding claims, comprising said electric sensor, which is an electro-optical or electro-chemical sensor, configured and arranged to detect microbial biofilms formed on the target surface (D ).
    [9]
    9.-Coupon holder (P) according to any one of the preceding claims, comprising said electrical transducer (Z) or an additional electrical transducer, which is an electro-thermal transducer configured and arranged to emit heat on said target surface ( D).
    [10]
    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 elongated 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 ]
    11. Coupon holder (P) according to claim 10, comprising two connection terminals (b) electrically connected with 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]
    12. Coupon holder (P) according to claim 10, comprising a local electric power generator disposed in the proximal end portion (1a) of the elongated body (1), and comprising respective output terminals connected to said seconds ends of said electrical conductors (W), said local electric power generator being configured to generate at least one electrical signal and provide it by said respective output terminals.
    [13]
    13.-Coupon holder (P) according to any one of the preceding claims, configured and sized for mounting, removably, in a head (H) of a biofilm reactor (R), by insertion into a through opening (Ap) thereof such that the distal end portion (1b) and the proximal end portion (1a) of said elongated body (1) remain, respectively, in and out of a vessel (V) of the biofilm reactor (R) on which the head (H) is placed.
    [14]
    14.-Coupon holder (P) according to claim 13, wherein said biofilm reactor
    (R) is of the CDC type.
    [15]
    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]
    16.-Biofilm reactor (R), comprising:
    - a vessel (V) to contain a culture medium under continuous agitation;
    - coupon holders (P) for microbial biofilms, where each one comprises an elongated body (1) with a proximal end portion (1a), of grip, and a distal end portion (1 b) 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), as a cover, and comprising through openings (Ap) for the passage and assembly, removably, of said coupon holders (P) ;
    characterized in that at least one of said coupon holders (P) is a coupon holder defined according to any one of claims 1 to 14.
    [17]
    17.-Biofilm reactor (R) according to claim 16. wherein the reactor (R) is of type
    CDC
    [18]
    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,
    - mount at least one coupon holder (P) for microbial biofilms defined according to any one of claims 1 to 14 on the support head (H), before or after coupling the support head (H) in the mouth of the vessel (V );
    - supply at least one electrical signal to the electrical transducer (Z) and / or electrical sensor disposed in the distal end portion (1 b) of the elongated body (1) of the coupon holder (P);
    - incubating the microbial inoculum for a certain time, subjected to continuous agitation within the culture medium, and maintaining said supply of said at least one electrical signal to the electrical 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]
    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 electro- 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 ]
    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 from thousands from kHz to tens of GHz.
    [21 ]
    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]
    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 of between 33 and 37 MHz.
    [23]
    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]
    24. Method according to any one of claims 18 to 23, wherein said microbial inoculum includes at least one clinical isolate of Candida albicans, Pseudomonas aeruginosa and Staphylococcus aureus, or a combination thereof.
    [25]
    25. Method according to any one of claims 18 to 24, wherein the reaction volume, culture medium plus microbial inoculum, is such that it does not cover the target surface (D), generating a humid chamber environment contaminated within of the vessel (V) of the biofilm reactor (R).
    [26]
    26. Method according to any one of claims 18 to 25, in said determined time is within a range from several hours to several days.
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    同族专利:
    公开号 | 公开日
    WO2018122442A1|2018-07-05|
    ES2677155B1|2019-05-08|
    引用文献:
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    ES201631733A|ES2677155B1|2016-12-30|2016-12-30|PORTA-COUPONS FOR MICROBIAL BIOPELICULES, SYSTEM AND REACTOR OF BIOPELICULAS THAT INCLUDES THE COUPON-HOLDER, AND METHOD TO OPERATE THE REACTOR|ES201631733A| ES2677155B1|2016-12-30|2016-12-30|PORTA-COUPONS FOR MICROBIAL BIOPELICULES, SYSTEM AND REACTOR OF BIOPELICULAS THAT INCLUDES THE COUPON-HOLDER, AND METHOD TO OPERATE THE REACTOR|
    PCT/ES2017/070869| WO2018122442A1|2016-12-30|2017-12-29|Coupon holder for microbial biofilms, system and biofilm reactor including said coupon-holder, and method for operating the reactor|
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