WO2016075561A1

WO2016075561A1 - Bag for the preparation of liquid culture media or broths

Info

Publication number: WO2016075561A1
Application number: PCT/IB2015/057553
Authority: WO
Inventors: Francesca PIEMONTI
Original assignee: Bioside S.R.L.
Priority date: 2014-11-11
Filing date: 2015-10-02
Publication date: 2016-05-19

Abstract

A bag (100) for the preparation of liquid culture media or broths comprises at least one compartment (10,40) containing a solid (101) and at least one empty compartment (20) suitable to contain a liquid (102). The compartments (10, 20, 40) are separated by at least one separation means (50) connectable with each other by at least one connection means (30) whose opening allow the solid (101) to come into contact with the liquid (102) and dissolve in it to form the broth. The solid (101) is a set of chemical components in defined concentrations and the liquid (102) is purified water or equivalent quality or an aqueous base solution. Preferably, the solid (101) is a powder or one or more tablets, or one or more capsules.

Description

DESCRIPTION

"Bag for the preparation of liquid culture media or broths"

[0001 ] This invention relates to a bag for the preparation of liquid culture media, in particular for the cultivation in the laboratory, transport, storage, dilution or suspension (for example Peptone Salt Solution), revitalisation (for example Buffered Peptone Water) , pre-enrichment and selective or^' non-selective enrichment (for example Tryptone Soya Broth, Fraser, Half Fraser, Buffered Peptone Water) of microorganisms.

[0002] The cultivation of microorganisms in the laboratory has the purpose of making available large quantities of biological material to adequately conduct laboratory investigations and for example allow practical applications in which micro-organisms are involved.

[0003] The microorganisms are made to reproduce on special substrates, defined culture media and fed with special nutrients .

[0004] The liquid culture media, commonly called broths, are constituted by a solution of various chemical components (synthetic and/or natural), in defined concentrations, in purified water or equivalent quality or in other aqueous base solutions. These media have a different composition in relation to the specific nutritional needs of the microorganisms cultivated.

[0005] An example of such culture media is Buffered Peptone Water .(ISO), which is to say a non-selective means of pre-enrichment for the detection of Salmonella in food samples and the like. Since Salmonella bacteria are often present in small numbers or are under stress, pre- enrichment gives the bacteria time to repair cells and multiply before being introduced in appropriate selective culture media, thereby improving the possibility of recovery from the sample to be analysed.

[0006] The careful preparation of culture media is one of the basic steps for ensuring the integrity of a microbiological examination. When dehydrated media are used as the basis for obtaining broths, the operations linked to their preparation must be done properly to avoid the manifestation of alterations of the components and microbial contamination. The steps necessary for the preparation of culture media are: weighing the chemical components, determination and addition of the volume of liquid (for example purified water or equivalent quality) , dissolution and dispersion, pH measurement and adjustment and sterilisation.

[0007] In order to speed up the preparation of culture media, avoiding at least the step of weighing the chemical components, it is known to use bags containing dehydrated media (in powder form) that are different depending on the type of microorganism to be cultivated. In this case, the laboratory technician determines the volume of liquid needed, pumps the liquid into the bag (through suitable connection tubes, filters, clamps and pumping systems) and dissolves the powder to obtain the broth. This type of operation exposes the broth to the risk of contamination during the step of pumping the liquid into the bag.

[0008] In order to eliminate any risk of external contamination, it is known to use bags containing broths ready for use. However, ready to use liquid media retains its characteristics for only a rather short time, rapidly alters and, in some cases, requires refrigerated storage temperatures.

[0009] The purpose of this invention is to provide a bag for the preparation of liquid culture media, or broths, able to solve the problems of the prior art.

[0010] In particular, the purpose of this invention is to provide a bag for the preparation of liquid culture media able to reduce the time needed for the preparation of the broth, avoiding the weighing of the chemical components, determination of the volume and addition of purified water, pH measurement and adjustment and sterilisation.

[0011] In addition, the purpose of this invention is to provide a bag for the preparation of liquid culture media able to eliminate any risk of external contamination.

[0012] Furthermore, the purpose of this invention is to provide a bag for the preparation of liquid culture media able to maintain its characteristics unaltered even for rather long times and without the need of refrigerated storage temperatures.

[0013] This purpose is achieved by a bag for the preparation of liquid culture media according to claim 1. The dependent claims describe preferred or advantageous embodiments of the bag.

[0014] The characteristics and advantages of the bag for the preparation of liquid culture media according to this invention will be apparent from the following description, given by way of non-limiting example, in accordance with the accompanying figures, wherein:

[0015] - Figure 1 is a perspective view of a bag for the preparation of liquid culture media according to this invention;

[0016] - Figure 2 is a top view of a bag according to this invention, according to a first embodiment;

[0017] - Figure 3 is a top view of a bag according to this invention, according to a further embodiment;

[0018] - Figure 4 is a top view of a bag according to this invention, according to a still further embodiment; [0019] - Figure 5 is a top view of a bag according to this invention, according to a still further embodiment;

[0020] - Figure 6 shows a detail of a bag according to this invention;

[0021] - Figure 7 shows a further detail of a bag according to this invention;

[0022] - Figure 8 is a top view of a bag according to this invention, according to a still further embodiment.

[0023] With reference to the accompanying figures, and in particular to Figure 1, a bag for the preparation of liquid culture media is shown, indicated by the reference number 100. For the bag, 100 is defined a longitudinal axis X.

[0024] The bag 100 is made of plastic material, preferably Polyethylene (PE), Polypropylene (PP), Polyvinylchloride

(PVC), Polyethylene terephthalate (PET), Ethylene vinyl acetate (EVA) or other synthetic and natural polymers.

[0025] Preferably, the bag 100 is made of mono-layer or multi-layer material.

[0026] Preferably, the bag 100 is made of plastic material, transparent or partially transparent or non-transparent, or partly or wholly coloured.

[0027] The bag 100 includes an upper end 110 and a lower end 120.

[0028] In a variant, shown in Figures 2 to 5, the bag 100 β

is obtained from a tubular body of flexible plastic welded in correspondence of the upper end 110 and lower end 120. Preferably, the weld is a substantially transversal weld 111.

[0029] In a further variant, shown in figure 8, the bag 100 is obtained from two sheets of plastic material, welded or glued along the perimeter 115.

[0030] Preferably, the bag 100 is adapted to be hung through the upper end 110.

[0031 ] In the variant embodiment of Figure 2, the upper end 110 extends for at least 2cm and is provided with a double weld 111. Such a construction makes the upper end 110 particularly strong and suitable to be inserted into specific support means, such as a gripper. Preferably, also the lower end 120 is provided with a double weld 111.

[0032] In the variant embodiments of Figures 3, 4 and 5, the upper end 110 is provided with at least one grip element 112,112'. Preferably, the grip element is a through hole 112 or through cut 112' made in the wall. Such construction makes the upper end 110 suitable to be hooked, or hung on special support means, such as hooks. Preferably, also the lower end 120 is provided with at least one grip element 112,112'.

[0033] In the variants of Figures 3 and 5, the upper end 110 is provided with three through holes 112 and of two cuts 112' , and the lower end 120 is provided with two cuts 112' .

[0034] In the variant of Figure 4, the upper end 110 is provided with a through hole 112.

[0035] The bag 100 comprises at least one compartment 10,40 containing a solid 101 and at least one compartment 20 suitable to be filled and to contain a liquid 102, and the compartments 10,20,40 are kept separated by at least one separation means 50 and are connectable through at least one connection means 30.

[0036] The solid 101 is a set of chemical components (synthetic and/or natural) at defined concentrations and the liquid is purified water or equivalent quality or an aqueous base solution.

[0037] Preferably, the solid 101 is a powder (as in Figure 1) or one or more tablets, or one or more capsules (as in figure 8 ) .

[0038] Preferably, the separation means 50 is welding or gluing. The separation means 50 defines compartments 10,20,40 at least partially in contour.

[0039] In the variant embodiment shown in Figure 2, the bag 100 comprises a compartment 10 containing the solid 101 and a compartment 20 containing the liquid 102, kept separated by a separation means 50. [0040] Preferably, the compartment 10,40 containing the solid 101 is placed above the compartment 20 containing the liquid 102.

[0041 ] In the variant embodiments of Figure 3, 4 and 5, the bag 100 comprises a compartment 10, a further compartment 40 and a compartment 20, kept separated by separation means 50.

[0042] In the variant embodiment shown in Figures 3 and 5, the compartment 10 and the further compartment 40 are kept separated from the compartment 20 by a first separation means 50 (by a first weld) and are kept separated from each other by a further separation means 50 (by a further weld) .

[0043] In the variant embodiment shown in Figure 4, the compartment .10 and the further compartment 40 are kept separated from the compartment 20 by a first separation means 50 (by a first weld) and are kept separated from each other by a by-pass duct 21, constituted by a portion of the compartment 20.

[0044] Preferably, the compartment 10 contains a solid, the further compartment 40 contains a further solid and the compartment 20 contains a liquid.

[0045] Preferably, the solids contained in the two compartments contain different chemical components.

[0046] In a variant, the compartment 10 contains a solid, the compartment 20 contains a liquid and the further compartment 40 contains a further liquid.

[0047] Preferably, the liquid contained in the two compartments is different.

[0048] In a further variant embodiment not shown, the bag 100 comprises a plurality of compartments containing a solid and/or a plurality of compartments containing a liquid .

[0049] Preferably, the compartment ( s ) containing a solid are placed above the compartment ( s ) containing a liquid. In further variants, the compartment ( s ) containing a solid are placed below or alongside the compartment ( s ) containing a liquid.

[0050] Preferably, each compartment 10, 20, 40 is provided with at least one filling channel 60, suitable to connect the compartment with the exterior of the bag 100.

[0051] Figure 7 shows in detail an embodiment of the filling channel 60, provided for example in correspondence of one end 110,120 of the bag 100. The filling channel 60 comprises a tubular body 61, provided with an inner lumen 62, and is placed astride between the compartment and the exterior of the bag 100. Preferably, the body 61 is welded in position through a weld, preferably the weld 111 provided at the ends 110,120.

[0052] In the variant embodiments of Figures 2, 3 and 5, the filling channel 60 of the compartment 10,40 containing the solid 101 is provided at the upper end 110 and the filling channel 60 of the compartment 20 containing the liquid 102 is provided at the lower end 120 of the bag 100.

[0053] In the variant embodiment of Figures 4, the filling channel 60 of the compartment 10,40 containing the solid

101 is provided at the upper end 110 and also the filling channel 60 of the compartment 20 containing the liquid

102 is provided at the upper end 120 of the bag 100. In this variant embodiment, wherein the compartments 10,40 containing the solid 101 are placed above the compartment 20 containing the liquid 102, the compartment 20 is provided with a by-pass duct 21, placed between the compartments 10 and 40, suitable to allow the outflow of the liquid from the upper end 110 of the bag 100 to the compartment 20.

[0054] Figure 5 shows a further embodiment of the filling channel 60, provided in correspondence of a wall of the bag 100. In particular, the filling channel 60 is a valve equipped with a cap 80, provided in correspondence of the compartment 20 containing the liquid. The valve is suitable to be connected to appropriate receiving means (for example needle free systems or other similar systems ) . [0055] During the production of the bag 100, the filling channel 60 allows filling, from the outside, each compartment with the related solid or liquid necessary for the preparation of the culture medium or broth. Once filled, the filling channels 60 are hermetically sealed with suitable closure means 80.

[0056] In the variant embodiment of Figure 5, the compartments 10,40 containing the solid are without filling channel. In this variant embodiment, during production of the bag, compartment 10,40 it is not fully closed (for example one side of the compartment remains open) in such a way as to allow easy loading of the solid. Once the filled, the bag is immediately welded (for example by means of the weld 111 at the upper end 110) to completely close the compartments 10,40 (now filled) and ensure the integrity of the contents.

[0057] Once filled with the solid component 101 (in a first production step) , and before filling with the liquid component 102, the bag 100 is subjected to sterilisation prior to marketing. Namely, the bag 100 in which the compartment 10 is filled with a solid 101 and the compartment 20 is empty (and thus the bag 100 contains no liquid 102) is subjected to sterilisation.

[0058] The sterilisation step is carried out for example by means of gamma irradiation and, more generally, by irradiation, or with moist heat or other sterilisation techniques .

[0059] Advantageously, the production method that provides for the sterilisation of the bag 100 in which the compartment 10 is filled with a solid 101 and the compartment 20 is still empty, allows considerably reducing transport and storage costs for performing the sterilisation .

[0060] When the sterilisation step is completed, the bag 100 containing only the solid component 101 a provided with a compartment 20 still empty, is ready for marketing. In this variant, therefore, the laboratory technician (i.e., the end user) will fill the bag 100 with the liquid component and, in particular, insert the liquid 102 in the compartment 20 through the filling channel 60.

[0061] In an alternative variant, in a second production step, the bag 100 (containing only the solid component 101) now sterilised is filled with the liquid component. Namely, the liquid element 102 is inserted into the compartment 20 through the filling channel 60. In this second production step, it is possible to use sterile raw materials and components and perform production in conditions such as to ensure the sterility of the finished product. [0062] When also the second production step is completed, the bag 100, now containing both the solid component 101 and the liquid component 102 (and thus provided with a filled compartment 20), is ready for marketing. In this variant, therefore, the bag 100 is ready for the laboratory technician (i.e., the end user) to use.

[0063] At the time of preparation of the culture medium, the laboratory technician will open the connection means 30 thereby allowing the solid 101 to come into contact with the liquid 102 and dissolve in it to form the broth.

[0064] In a variant embodiment, shown in Figures 2 to 4, the connection means 30 is a valve. Figure 6 shows in detail the connection means in the form of a valve. The valve comprises a fixed part 31, equipped with an inner lumen 33, located astride the compartments to be connected. Preferably, the fixed part 31 is welded in position through a weld, preferably the weld that defines the separation means 50.

[0065] The valve comprises an openable portion 32, positioned to close the inner lumen 33, suitable to be opened to form the connection between the compartments. In the variant of Figure 6, the openable portion 32 is suitable to be detached, separated from the valve to form the connection between the compartments. In a variant, the openable portion 32· is suitable to be broken to form the connection between the compartments. In a further variant, the openable portion 32 is a cap inserted in the lumen 33 and suitable to be removed to form the connection between the compartments.

[0066] In the variants shown in Figures 2 to 4, the laboratory technician will break the seal created by the valve 30, thus allowing the solid 101 to come into contact with the liquid 102 and dissolve in it to form the broth.

[0067] In a variant embodiment, shown in Figure 5 and 8, the connection means 30 is a yieldable weld or gluing realised between the two walls of the bag 100. Preferably, the yieldable weld is the weld that defines the separation means 50 between the compartments. In this variant, the yieldable weld or gluing is suitable to be opened, unglued, to form the connection between the compartments .

[0068] In the variant shown in Figures 5 to 8, the laboratory technician will break the seal created by the yieldable weld or gluing and, in particular, separate the two walls of the bag previously joined to each other, thus allowing the solid 101 to come into contact with the liquid 102 and dissolve in it to form the broth.

[0069] In another variant embodiment, the connection means 30 are positioned both between the compartments 10,40 and the compartment 20 and between the compartment 10 and the further compartment 40.

[0070] The bag 100 comprises at least one emptying channel 60, suitable to allow the escape of the broth to the outside of the bag 100. Preferably, the emptying channel 60 is positioned in correspondence of the compartment 20 containing the liquid. Preferably, the emptying channel 60 is provided at the lower end 120 of the bag 100.

[0071 ] Preferably, the emptying channel 60 is the filling channel 60.

[0072] Preferably_? the emptying channel 60 is provided with a tubular body 61 and an inner lumen 62 (as in Figure 7) or is a valve with a cap (as in Figure 5) .

[0073] After the culture broth is obtained, the laboratory technician will put the emptying channel 60 in connection (for example by piercing the cap 80 with special sterile spikes, needle free systems or other similar systems) with adequate receiving means, thus allowing the escape of the broth from the bag 100 and its use for the cultivation of microorganisms in the laboratory.

[0074] Preferably, the bag 100 comprises at least two emptying channels 60 (as shown in Figures 2, 3 and 8), each provided with a cap 80. This variant embodiment allows drawing the broth ready in the bag 100 at a later moments: in a first moment, the laboratory technician will put the first emptying channel in connection with suitable receiving means, to draw a portion of the broth from the bag, and to close the first emptying channel by means of, for example, a clamp; in a second moment, the laboratory technician will put the second emptying channel (that remained intact and sealed) in connection with suitable receiving means, to draw a further portion of the broth from the bag.

[0075] The bag 100 is suitable for preparing at least one litre of broth. Preferably, the bag 100 is suitable to be produced in different sizes: 5 litres, 4.5 litres, 3 litres, 2.25 litres and 1 litre. The bag 100 is also suitable to be produced in further sizes: 7.5 litres, 10 litres, 15 litres and 20 litres.

[0076] In an example embodiment, the bag 100 is suitable for the preparation of a pre-enrichment medium to be used for the isolation of Salmonella and Shigella from foods. In this embodiment, the solid element is a powder that contains the following components, unit of measurement g/litre of distilled water: Peptone (10.0), Sodium chloride (5.0), sodium phosphate monoacid (3.5), potassium dihydrogen phosphate (1.5). PH 7.2 ± 0.2.

[0077] Advantageously, a bag comprising at least one compartment containing a solid and at least one empty compartment suitable to be filled and contain a liquid allows reducing transport and storage costs due to its small size. Advantageously, the use of bags comprising at least one compartment containing a solid and at least one empty compartment suitable to be filled and contain a liquid gives the laboratory technician the possibility of performing, in a single loading session (for example weekly, monthly, quarterly, etc.), to fill the compartment for the liquid of several bags on site. In this way, laboratories can use only one filter, one time to fill "n" bags of small size. This is possible due to the fact that, in this variant of the bag, loading of the water does not correspond to the reconstitution of the broth, but only to the step of filling the compartment for the liquid. Filling the compartment for the liquid makes it possible to complete the bag in the laboratory, which until then, was equivalent to an intermediate processing. This solution completely solves the negative factor of the cost of the filter, which is fully amortised since used to fill a large number of bags in a single session. For example, it is possible to load 180 litres of liquid, divided into 60 bags of 3 litres each, rather than 36 bags of 5 litres each. Furthermore, since the broth is still to be reconstituted, the bags are maintained long-term at room temperature. Advantageously, loading the liquid directly in the laboratory and that does not involve the reconstitution of the broth allows reducing the critical contamination points (the loading can be carried out in special sessions scheduled periodically) , to reduce the costs of the capsule filter (a single filter is used for filling several bags), to obtain bags of manageable volumes in terms of daily consumption even by small laboratories, to obtain a bag that is stable and sterile over time, since the broth is yet to be reconstituted.

[0078] Innovatively, a ready to use bag for the preparation of liquid culture media according to this invention allows to considerably reducing broth preparation times.

[0079] Advantageously, a ready to use bag for the preparation of liquid culture media according to this invention allows completely eliminating the risks of external contamination in the broth preparation step.

[0080] Advantageously, a ready to use bag for the preparation of liquid culture media according to this invention allows can be stored for long times, even at room temperature, without altering its characteristics.

[0081] Advantageously, a ready to use bag for the preparation of liquid culture media according to this invention is particularly suitable for the preparation of broths for use in the food, environmental, cosmetic, pharmaceutical, veterinary and other industries and in other microbiological diagnostic contexts.

[0082] Advantageously, a ready to use bag for the preparation of liquid culture media according to this invention does not require: the addition of water and other reagents; sterilisation before use (sterilisation of the finished product is performed (or assured) during production by irradiation or other techniques suitable for the purpose and avoids the risk of overheating and the loss of performance) ; refrigerated storage (with consequent energy savings, during both transport and storage); of aseptic procedures in the laboratory (thanks to the use of a dual output channel equipped with seals acts to ensure sterility; manipulation of glass objects (with a consequent increase in operator safety) .

[0083] Advantageously, a ready to use bag for the preparation of liquid culture media according to this invention can be manufactured in various easy to handle sizes.

[0084] It is clear that one skilled in the art, in order to meet contingent needs, may make changes to the bag described above, all contained within the scope of protection defined by the following claims.

Claims

1. Sack (100) for the preparation of liquid culture media or broths, comprising at least one compartment (10, 40) containing a solid (101), and at least one empty compartment (20) suitable to be filled with a liquid (102), said compartments (10, 20, 40) being separated by at least one means of separation (50) and connectable by at least one means of connection (30), wherein, when the compartment (20) is filled with liquid, the opening of the connection means (30) allows the solid (101) to come into contact with the liquid (102) and dissolve to form the broth.

2. Sack (100) according to claim 1, wherein the means of separation (50) is a weld or gluing which at least partly defines the contour of the compartments (10, 20, 40) .

3. Sack (100) according to claim 1 or 2, wherein the connection (30) is at least a yielding weld or gluing provided between the two walls of the sack (100), suitable to be opened or separated to make the connection between the compartments.

4. Sack (100) according to any of the previous claims, comprising an upper end (110) and a lower end (120), wherein the upper end (110) and/or the lower end (120) is suitable to be attached or hung on appropriate support means by at least one grip element (112, 112') ·

5. Sack (100) according to claim 4, where the grip element is a through-hole (112) or a through incision (112') made in the walls of the sack (100) .

6. Sack (100) according to any of the previous claims, wherein the compartment (10, 40) containing the solid (101) is placed above the compartment (20) for the liquid (102) .

7. Sack (100) according to any of the previous claims, comprising at least an additional compartment (40) for a further solid or liquid.

8. Sack (100) according to any of the previous claims, wherein the compartment (10, 20, 40) has at least one filling channel (60) suitable to connect it with the outside of the sack (10) for the filling of the compartment (10, 20, 40) .

9. Sack (100) according to claim 8, wherein the filling channel (60) of the compartment (10, 40) containing the solid (101) is provided at the upper end (110) and the filling channel (60) of the compartment (20) for the liquid (102) is provided at the lower end (120) of the sack (100) .

10. Sack (100) according to claim 8, wherein the filling channel (60) of the compartment (10, 40) containing the solid (101) is provided at the upper end (110) and the filling channel (60) of the compartment (20) for the liquid (102) is also provided at the upper end (110) of the sack (100) .

11. Sack (100) according to claim 10 when dependent on claim 7, wherein the compartments (10,40) containing the solid (101) are placed above the compartment (20) for the liquid (102), and the compartment (20) for the liquid (102) is fitted with a by-pass duct (21), placed between the compartments (10,40) containing the solid (101), suitable to allow, during filling, the passage of liquid (102) inside the compartment (20).

12. Sack (100) according to any of the claims 1 to 10, wherein the compartment (10, 40) containing the solid

(101) is devoid of any filling channel (60).

13. Sack (100) according to any of the previous claims, comprising at least one emptying channel (60), placed at the compartment (20) for the liquid (102), suitable to permit the output of the broth to the outside of the sack (100) .

14. Sack (100) according to claim 13, wherein the emptying channel (60) corresponds to the filling channel (60) .

15. Sack (100) according to claim 13 or 14, comprising two emptying channels (60).

16. Sack (100) according to any of the previous claims, made of plastic material, preferably in polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), ethylene vinyl acetate (EVA) or other synthetic and natural polymers.

17. Sack (100) according to any of the previous claims, where the compartment (20) contains a liquid (102), and the liquid (102) is purified water or of equivalent quality or an aqueous solution.

18. Sack (100) according to any of the previous claims, wherein the solid (101) is a collection of chemical components in defined concentrations, and is a powder, or one or more tablets or one or more capsules.

19. Production method of a sack (100) according to any of claims 1 to 18, comprising the steps of:

- fill the compartment (10, 40) with a solid (101),

- sterilize the sack (100) before the filling of the compartment (20) with a liquid (102), in such a way that the sack (100) is sterilized when the compartment (20) is still empty.

20. Production method of a sack (100) according to claim 19, comprising the further step of:

- fill the compartment (20) with a liquid (102) by using sterilized tools.