HU200877B - Method and apparatus for sterile and/or partially sterile preparing, grafting and filling-down nutrient substratum of mushroom - Google Patents

Abstract

The present invention relates to a sterile or partially sterile preparation, inoculation and downloading of a fungus medium, wherein the known base material is a straw. 5 mm and / or corn cob. Cut to less than 6 mm, grind, and mix with the necessary additives, preferably ammonium nitrate. The thus prepared material is wetted by the addition of water, the wetted material is germinated by partial sterilization by heating and fermentation. The fungus medium prepared in this way is inoculated, transformed and placed in a growing area. The process is characterized in that the prepared feedstock is added to a mixing vessel closed with additives known per se, in a mixing blender having a rotation of 40-80 rpm, preferably 60 rpm, with stirring at least 60%, preferably 70%. until the moisture content is reached, the material is moistened in two stages by adding water at a temperature of at least 90 ° C in the first stage, and in the second stage to a final moisture increase of 7-10%, preferably 7.5%, at a temperature of 100-120 ° C. , Scope of description: Page 8, Figure 1 -1-

Description

Preferably, this is achieved by adding saturated water vapor at 110 'C for 5 to 30 minutes, preferably 15 minutes. The material is sterilized by steam at 100-120 ° C, preferably 115 ° C, for 30-60 min, preferably 35 min, by external heating of the closed mixing vessel. While maintaining agitation, the material is cooled to a temperature of 20-30 ° C, preferably 25 ° C, by cooling the outer surface of the closed mixing vessel while equalizing the internal pressure in the closed mixing vessel to atmospheric pressure. In a sealed mixing vessel, a particle size of 1-3 mm, preferably 1.5 mm, is delivered from the germ-free space with which the substance is further stirred for at least 5 minutes. The mixture is added to sealed sterile containers for germination and / or cultivation in sealed space and sealed.

The invention further relates to an apparatus for preparing, inoculating, or loading sterile or partially sterile fungal medium.

The device is characterized by a hermetic inner space (5) with a lid (2), a narrowing extension jacket (3) on the side, and an openable or hermetically sealed vaccine lid (5) arranged at an upper point of the hermetic interior (5). 9) equipped with a vaccine inlet opening (7) and a raw material inlet (6) with a hinged and hermetically sealed raw material cover (6) and an openable or hermetically sealed dispensing cover (28) on the underside of the narrowed jacket (3) has a medium dispensing opening (27). The cover (2) has a conveyor mixing element (12) pivotally mounted, hermetically sealed and mounted on a shaft (11), connected to a gear unit (10). A steam-water inlet (16) is provided at the hermetic interior (5). A cylindrical jacket (4) enclosing a hermetic inner space (5) with a tapered extension jacket (3) is surrounded by a heating or refrigerant flow outer jacket (18) having a fuel inlet coolant outlet (s) (20) and a coolant inlet coolant (20) an outlet arrow (s) (21) are provided. (Figure 1)

-2HU 200877 Β

The present invention relates to a process for the preparation, inoculation and loading of sterile or partially sterile fungal medium, wherein the known raw material, preferably straw, is passed through. 5 mm and / or corn cavity diameter. It is cut to size smaller than 6 mm, minced, mixed with the necessary additives, preferably ammonium nitrate, then moistened with water, germinated by heating, fermentation, partial sterilization, inoculated and stained.

The invention also relates to apparatus for preparing, inoculating and loading sterile or partially sterile fungal medium.

It is known that certain fungal species, e.g. It can also be grown on oyster mushrooms, timber, cereals and various agricultural by-products. Various methods of preparing the culture media required for cultivation are known. The purpose of each preparation method is to create favorable living conditions for the fungal mycelium to be cultivated. In doing so, the mushroom medium contains the nutrients and water needed for the mushroom. The preparation also aims to eliminate many of the competing microorganisms (molds, bacteria) in the raw material from fungal medium, or to reduce or eliminate their number. Thus, the fungal mycelium entering such a fungal medium can grow rapidly without competition. Thus, the fungal medium can be prepared from dry raw materials by moistening and then eliminating the harmful effects of competing microorganisms. The woven material is either used for further grafting or used on the crop for mushroom production (fruiting).

Three methods of heat treatment to reduce harmful microorganisms following wetting are known and are commonly used. The first method of heat treatment is to propagate thermophilic bacteria into the culture medium to provide the necessary protection. During the heat treatment, the wetted raw material is max. It is heated to 60'C - against insect pests, possibly for 2-3 hours to 70'C, but then cooled to 60'C and kept for 2-3 days at 50-55 with proper aeration. At this point, the proliferation of very high levels of thermophilic bacteria in the material will target the nutrients of molds and other microorganisms, which will result in some protection in the medium. In this method, the wetted raw material can be heated by composting, steam, immersion in hot water or so-called mass heat treatment. The heat-treated medium can be woven and used after being cooled to below 30 ° C, under no particular hygiene conditions, inoculated with fungal mycelium and then filled into containers. The procedure for this method can be found in Sándor Balázs's "Mushroom Cultivation" (Agricultural Publisher, 1979), pp. 59-60. side.

In the second heat treatment method, during the partial sterilization process, the wetted raw material is heated to a temperature of 70-100 ° C and maintained for 1-6 hours. At this point, the vermin and the vegetative forms of competing microorganisms in the raw material are destroyed, but the more resistant forms of spores are not. After cooling, the medium is rapidly cooled to below 30 ° C and inoculated with the mycelium of the selected fungus. Strict hygiene conditions must be ensured at the time of inoculation, since partial sterilization of the culture medium does not provide protection against spores in the form of spores and infectious organisms from air or other sources. After inoculation and filling into beads, strict hygienic conditions must be maintained during the transfusion. Therefore, the vaccine should be sealed after vaccination to ensure proper germination of the container. The woven fabric already protects itself from infections. Thus, the interweaving may occur under natural - normal - conditions.

The third method of heat treatment is the sterile method. Here, the wetted raw material is filled and sealed in a heat-resistant container, then heated together with the container in an autoclave under pressure to a temperature of 110-130 ° C. This temperature kills pests and all microorganisms (molds, spores, bacteria) in the raw material. The medium is then cooled to a certain temperature in the autoclave, and then the containers are cooled below 30 ° C except for the autoclave. The medium is then inoculated with fungal mycelium under very strict hygiene conditions and then sealed again. The medium is then woven in a suitable location and used for cultivation. Mostly, this method produces only vaccines.

As is known, the following methods and devices are used for the technical implementation of the preparation, inoculation and download of the sterile medium.

The crushed or small-grained (wheat grain) raw material is first moistened or pre-cooked to ensure adequate water uptake. Wetting is usually performed by manual force, infusion and mixing, using large quantities of conveyors (belts). A variety of batch and continuous cooking equipment is used to pre-cook the grain raw material. The pre-wetted and cooked material, after cooling and desiccation, is usually filled by hand into containers, most often canned jars, and then sealed by hand in a germ-poor closure. Filled bottles are autoclaved by hand or basket lifter, sterilized at appropriate pressure and temperature. After being kept cool, the filled bottles are allowed to cool to a certain temperature in the autoclave and then, except for the autoclave, disintegrated in a place where it is naturally cooled to cool to the extinguishing temperature. Then, under sterile conditions, the vials are manually reopened and the inoculum or inoculum is manually inoculated with the sterile medium and the vials are closed again by hand. The medium is then transfixed at the appropriate location. Such procedures are described in Sándor Balázs' s "Our Cultivated Mushrooms" (Akadémiai Kiadó, 1982), pp. 99 and 245-246. pages.

The near sterile mode heat treatment of the fungal medium can also be carried out in an autoclave in smaller amounts, just like the previous ones, but at lower temperatures. Vaccination and transfusion are both identical to those described for the sterile method. For larger quantities, insulated steaming he3

EN 200877 Β the humidified raw material filled in boxes or foil bags is steam-heated to the desired temperature and then kept in heat. It is then cooled as rapidly as possible by the air flow. The heat-treated medium is then removed from the steamer. The vaccine is poured into a machine or 5 hands to mix the vaccine, then filled again in a crate or bag. Inoculation may also be carried out by inserting a dipstick into the container used with the steamer or by manual force inoculation. After inoculation, the packages are sealed with a germ-less closure. In both cases, vaccination is carried out under strict hygiene conditions. The medium is then woven under conditions of enhanced hygiene (Sándor Balázs 'Our Cultivated Mushrooms', Akadémiai Kiadó, 1982, p. 249).

A disadvantage of the known and hitherto used methods and devices is that only certain stages of operation are known in the preparation, inoculation and loading of both sterile and near sterile mushroom media. A further disadvantage is that each operation stage has so far been implemented in 20 separate rooms with separate equipment. Accordingly, these operations required a large amount of material handling and manual workflows. A further disadvantage of the methods used, which is related to the infectious sensitivity of the heat-treated medium from the environment, is that there is an increased risk of infection at all stages of the operation - reloading, unfolding. A particular disadvantage of autoclave processes is that due to unfavorable heat transfer conditions, only small units (about 5 liters) can be heat treated, but also with disproportionately high energy consumption. Cleaning the bottles before they are reused also causes additional costs. A further disadvantage of the methods used is that the large amount of manual labor and handling of the necessary material handling tasks adversely affect the costs and the economics of these methods. These disadvantages are more pronounced in large-scale production. Therefore, the sterile process on an industrial scale was only economical for the production of fungal germ. Large-scale preparation, inoculation and retrieval of 40 culture media for mushroom cultivation could not be accomplished by either sterile or partially sterile methods without proper procedures.

SUMMARY OF THE INVENTION It is an object of the present invention to overcome the above disadvantages by providing a process and apparatus for preparing, inoculating and loading sterile and partially sterile fungi medium, which involves or provides raw material moistening, heat treatment, medium inoculation and loading operations with low material handling it can be done with minimal work effort, reducing the risk of medium contamination, thereby ensuring economical, large-scale plant applicability.

The present invention therefore relates to a process for preparing, inoculating and loading sterile or partially sterile fungal medium, wherein the known raw material is straw. 5 mm and / or corn cavity diameter. Cut to size smaller than 6 mm, minced, mixed with the necessary additives, preferably ammonium nitrate, then moistened with water 60, germinated by heating, fermentation, partial sterilization, inoculated and strain disposed in such a manner that the prepared feedstock is added to a closed mixing vessel with additives according to the well-known re4 formulations in which it is stirred at 40-80 rpm, preferably 60 rpm, with an agitator-agitator operating at least 60%, preferably 70 until the moisture content is reached, the material is wetted in two stages by adding water at a temperature of at least 90 ° C in the first stage, and in the second stage by a final increase in humidity of 7-10%, preferably 7.5%, 100-120 '. C temperature is achieved by adding a saturated water vapor needle, preferably 110 ° C, for 5 to 30 minutes, preferably 15 minutes, followed by 100-120 ° C, preferably 115 ° C, steam. and sterilized by external heating of a small bowl for 30-60 min, preferably 35 min. While maintaining the agitation, the material is cooled to 20-30 ° C, preferably 25 ° C, by means of a refrigerant applied to the outer surface of the closed mixing vessel while equalizing the internal pressure in the closed mixing vessel to atmospheric. From the germinated space, 1-3 mm, preferably 1.5 mm, of inoculant is delivered to the closed mixing vessel for further agitation for at least 5 minutes, and the mixture is added to the sterilized space for germination and / or cultivation in closed sterile containers and is closed.

The invention also relates to apparatus for preparing, inoculating and loading sterile or partially sterile fungal medium. A hermetic inner space of the apparatus surrounded by a cap, a cylindrical jacket enclosed by a narrowing extension jacket, which is an opening and a hinged opening for a vaccine, a hinged opening and a hinged opening, a hinged opening and a hinged opening has a media outlet with an opening and hermetically sealed dispensing lid, a motor-connected, rotatably mounted hermetically sealed shaft, a hermetically sealed inner and auxiliary fluid, and a water inlet 45 is provided, and the welded jacket having a tapered extension jacket to limit the hermetic interior is flowed by a heating or cooling medium The lake is surrounded by an outer jacket on which the fuel inlet coolant outlet (s) and the refrigerant inlet fuel outlet (s) are (are) formed.

In a preferred embodiment, the cylindrical sheath has a hermetically sealed scraper axle pivotally mounted on the longitudinal axis of the cylindrical housing on which the scraper blades and the scraper disc are fixed.

It may also be typical that the feed inlet is provided with a feed conveyor and the vaccine inlet is provided with a feed conveyor.

It is also possible that an air outlet is provided on the underside of the tapered extension jacket, preferably an aseptic pressure equalizer connected to the air intake duct in the lid.

In a further preferred embodiment, the inlet 65 is provided with a hopper in which

EN 200877 Β a rod shaft is located above the vaccine conveyor and the hopper is fitted with a hinged opening and a hermetically sealed closure.

It is also possible that, preferably, a cleaning line is connected through the cover, on which nozzles are arranged in the airtight interior.

In a further preferred embodiment, a vapor vapor conduit and a condensate conduit are connected to the air intake conduit.

The process and apparatus of the invention have numerous advantages. The unit is capable of performing the process of humidification, heat treatment, extinguishing and downloading in a single hermetic space. This minimizes the risk of medium contamination compared to known methods. Due to the simple construction of the equipment, it can be produced at favorable investment costs. The process can also be implemented with enhanced automatic control of the equipment so that it can be operated with little labor. The duration of each work phase can be kept constant regardless of the operator. The operations are performed in a single space. Intermediate material handling is completely absent. The plant can be built for large volume processing of several m ^{3 of} raw materials. Because of its good space requirements, strict hygiene conditions need to be ensured in only a small part of the building, which reduces the additional investment costs. All this makes the process and the equipment advantageous for large-scale production.

However, the inventive device has the disadvantage that it can only be manufactured in a well-equipped manufacturing plant, as compared to conventional heat treatment devices, which require less technical preparation.

The present invention will now be described in more detail by way of example.

Example

In this example, 100 kg of Pleurotus ostretus (oyster mushroom) was prepared. 35 kg corn cobs diameter Grind to a particle size of 5 mm and add to a closed mixing vessel. The material is stirred in the apparatus at 60 rpm. Subsequently, 57 kg of water at a temperature of 95 ° C is added over a period of 10 minutes. This gives the first degree of wetting. Subsequently, 5 kg of saturated water vapor at 110 ° C is introduced into the apparatus. During this process, this amount of vapor is "condensed" into the materials there (thus providing the second stage of wetting!). After the second wetting step, begin the sterilization process, which is indirectly provided to prevent the further moisture from entering, by maintaining the temperature required for sterilization through heat transfer through the outer wall of the device. The heat retention process is maintained for 30 min after reaching an internal temperature of 100 ° C while stirring the material evenly. Once the heat is complete, the mixture is cooled (and heat is removed through the outer wall of the apparatus) while stirring is continued. This process is maintained until the internal temperature reaches 25 'C. Subsequently, the mixing vessel, under strict hygienic conditions, weighs 5 kg

Pleurotus ostreatus mushroom sprouts are added in finely crushed state with continuous mixing. Stir the inoculated material evenly for 10 min. Stirring is then stopped and the inoculated material is emptied into sterile bags under strict hygiene conditions. The bags are sealed with an airtight seal and then placed in a 25 ° C permeation space.

The apparatus according to the invention will be described in connection with an exemplary embodiment by way of a drawing. 1 is a schematic sectional view of an assembly of the apparatus of the present invention. The hermetic interior 5 of the apparatus is defined by a lid 2 from above, a tapered extension jacket 3 from below, and a cylindrical jacket 4 from the side. At one of the upper points of the hermetic interior 5, on the cylindrical sheath 4, the raw material inlets 6 and the vaccine inlets 7 are formed. The inlet openings 6 are closed by an openable or hermetically sealed stock cover 8. The vaccine inlet opening 7 is closed by an openable or hermetically sealed vaccine lid 9. On the underside of the tapered sheath 3 there is provided a media dispensing opening 27 with an openable or hermetically sealed dispensing cover 28. In the longitudinal axis of the cylindrical sheath 4, the hermetically sealed shaft 11, which is hingedly sealed in the lid 2 and engages with a gear 10 from above, extends into the hermetic interior. The shaft 11 is provided with a conveyor mixing element 12, in this embodiment a screw. A vapor-water inlet port 16 is formed in the cylindrical jacket 4 through steam-water closure fittings 17 connected to the airtight interior 5. A cylindrical jacket 4 with a tapered extension jacket 3 is surrounded by an outer jacket of fuel or coolant stream 18, which is provided with coolant inlet coolant outlets 20 and coolant inlet coolant outlet openings 21. Between the cylindrical jacket 4 with the tapered solution 3 jacket and the outer jacket 18 there is a jacket space 19. A hermetically sealed scraper shaft 15, which is coupled to a centerline of the cylindrical jacket 4, is mounted on a hermetically sealed scraper 13 and has a plurality of scraper blades 14 and a scraper disc 34 thereon. A feed conveyor 47 is arranged in the feed hopper 47 for the feed inlet 6. The vaccine inlet 7 is provided with a vaccine conveyor screw located in a hopper 48. Above the vaccine transport screw 31, a shaft 32 is disposed in the hopper 48. The hopper 48 is covered by a closure 33 which can be opened or hermetically closed from above. An air outlet 24 is provided at the lower end of the tapered extension jacket 3 and is provided with an air closure assembly 25. An aseptic pressure equalizer 35 is formed in the lid 2 via an air inlet conduit 54 through the venting closure 50. The cover 2 passes through a cleaning line 37 provided with a cleaning fluid closure assembly 38, in which nozzles 36 are arranged in the hermetic interior 5.

In the case of cooling, a vapor barrier 49 with a vapor barrier closure 51 is connected to the air intake duct 54 and a condensate conduit 55 with a condensate barrier duct 52 to the purification line 37. The fuel supply fluid 20 is 5

EN 200877 22 22 locking fittings are connected to the outlet. A conduit 53 is connected to the refrigerant inlet and outlet outlet 21 via valve 23. The thermometer sensor 46 extends into the hermetic interior 5. The outer sheath 18 and the cover 2 are covered by a heat-insulating layer 26. On the outside, 1 scaffold is connected to the tapered sheath 3. When the dispensing lid 28 is open, a container 29 is connected to the medium dispensing opening 27.

The operation of the apparatus according to the invention may be summarized as follows. The raw material is conveyed into the feed hopper 47 of the feed conveyor 30 from where the feed conveyor 30 is transported through the feed inlet 6 into the airtight interior 5 with the feed lid 8 open and filled to about two-thirds volume. The raw material inlet opening 6 is hermetically sealed with the raw material cover 8. The gear 10 and the scraper 13 are then started. The scraping gear 13 rotates the scraping strips 14 connected thereto through the scraping shaft 15. The pitch and direction of rotation of the conveyor mixing element 12 is such that it conveys the raw material upwards, which then slides downwardly along the tapered extension jacket 3 into something. This results in a continuous mixing of the raw material. The material adhering to the surface of the hermetic inner space 5 of the cylindrical jacket 4 and the tapered extension jacket 3 is continuously scraped off by the scraping strips 14. The air shut-off valve 25 is opened, and then the steam-water shut-off valve 17, through which hot water is injected into the hermetic blister space 5. The vapor of the incoming hot water displaces the air there and exits through the air outlet 24. The air closure assembly 25 is sealed and saturated steam is introduced through the vapor-water closure assembly 17 into the airtight interior 5 to achieve the desired degree of humidity. For sterilization, saturated steam is also introduced through the closure assembly 22 into the jacket space 19, the condensate of which is discharged through the refrigerant inlet and outlet 23 and the valve 23. The temperature of the hermetic material 5 is shown by the thermometer 46. Cooling of the material is provided by the cooling water discharged through the valve 23 and the refrigerant inlet and outlet 21 and the closure 22. As a result of the cooling, the depression created in the hermetic inner space 5 is compensated by the aseptic pressure equalizer 35 to the pressure of the outer space. The injection of the vaccine into the airtight inner space 5 is carried out by the vaccine conveying screw 31 via the hopper 48 after the closure has been lifted. The pre-shredded vaccine will enter into the hermetic space 5 and will be sufficiently fine through the crushing wheel. After mixing the vaccine with the fungal medium, the drive 10 and the scraper 13 are stopped. To lower the inoculated material into the container 29, the dispensing closure 28 is opened and the container 29 is placed under the medium dispensing opening 27. Subsequently, both the gear unit 10 and the scraper gear 13 are rotated in opposite directions, whereby the conveyor mixing element 12 forces the material towards the container and the material uniformly mixed with the vaccine is discharged into the container 29. After emptying, the hermetic interior 5 is flushed with the cleaning fluid introduced through the cleaning line by opening the closure of the cleaning fluid 38 and scattered by the nozzles 36.

If vacuum cooling is desired for quicker cooling of the material, the evacuated vapors may be removed by opening the vapor barrier assembly 51, the condensate of which is returned through the nozzles 36 through the nozzles 36 to the hermetic interior 5.

Claims (8)

A process for the preparation, inoculation and loading of sterile or partially sterile fungal medium, wherein the known raw material is straw; 5 mm and / or corn cavity diameter. It is cut, minced and then mixed with the necessary known additives, such as ammonium nitrate, moistened with water, germinated by heating, fermentation, partial sterilization, inoculated, placing the material into a growing space, characterized in that the prepared feedstock is added to the mixing vessel in a closed mixing vessel with additives according to known formulas, which are stirred at 40-80 rpm, preferably 60 rpm, with at least 60%, preferably To reach a moisture content of 70%, the material is wetted in two stages by adding water at a temperature of at least 90 ° C in the first stage and then, in the second stage, to the final stage. A 7-10%, preferably 7.5%, increase in humidity is achieved by adding saturated water vapor at 100-120 ° C, preferably 110 ° C, for 5 to 30 minutes, preferably 15 minutes. , the material is sterilized by steam at 100-120 ° C, preferably 115 ° C, for 30-60 min, preferably 35 min, by external heating of the closed mixing vessel, followed by cooled to a temperature of 20-30 ° C, preferably 25 ° C, by cooling the outer surface of the closed mixing vessel while equilibrating the internal pressure in the closed mixing vessel to atmospheric pressure, followed by 1-3 mm, preferably 1.5 mm, of germ-free space in the closed mixing vessel , delivering a granule-sized vaccine with which the substance is further agitated for at least 5 minutes and then the mixture is germinated, and sealed. Apparatus for preparing, inoculating and loading sterile or partially sterile mushroom medium, characterized in that it has a hermetic inner space (5) surrounded by a lid (2), a cylindrical sheath (3) bordered by a tapering extension (3). 5) at an upper point EN 200877 Β a vaccine inlet (7) with openable or hermetically sealed vaccine cover (9), and a feed inlet (6) with an openable or hermetically sealed access cover (8) and a narrower extension sleeve (3) having a medium dispensing opening (27) with a hermetically sealed dispensing lid (28), a conveyor element (10) connected to the gear (10) on the lid (2) and mounted on a rotatably mounted hermetically sealed shaft (11) 12) and a vapor-water inlet (16) connected to the hermetic interior (5) and a heating or cooling medium flow for the cylindrical jacket (4) with a tapered jacket (3) delimiting the hermetic interior (5) enclosed by an outer jacket (18) having a fuel inlet (s) (20) and a refrigerant inlet fuel outlet (s) (21) are provided. Apparatus according to claim 2, characterized in that the cylindrical jacket (4) has a hermetically sealed scraper shaft (15) which is pivotally mounted on the longitudinal axis of the cylindrical casing and has a scraper bar (14) and a grinding wheel (34). recorded. Apparatus according to claim 2 or 3, characterized in that a feed screw (30) is connected to the feed inlet (6) and a feed screw (31) to the vaccine inlet (7). . 5. Apparatus according to any one of claims 1 to 3, characterized in that an air outlet (24) is formed on the underside of the tapered extension jacket (3), preferably an aseptic pressure equalizer (35) connected to the air intake duct (54). 6. Apparatus according to any one of claims 1 to 3, characterized in that a hopper (48) is connected to the vaccine inlet (7), in which a rod shaft (32) is arranged above the vaccine transport screw (31) and the hopper (48) can be opened or hermetically provided with a lockable locking member (33). 7. Apparatus according to any one of claims 1 to 3, characterized in that nozzles (36) arranged in the hermetic interior (5) are connected to the cleaning line (37). 8. Apparatus according to any one of claims 1 to 3, characterized in that a vapor conduit (49) is connected to the air inlet conduit (54) and a condensate conduit (55) is connected to the purification conduit (37).