A microbial composition, a microecological preparation and its application in breeding sea cucumbers

Abstract

The present invention belongs to the field of microorganisms, and relates to a microbial composition, a microecological preparation and an application thereof in raising sea cucumbers. The microbial composition comprises Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris, wherein the ratio of the number of live bacteria of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1:(0.1-10):(0.1-10). Raising sea cucumbers with the microbial composition provided by the present invention can effectively improve the indicators such as fatness, skin emergence rate, body wall thickness, wart foot height and muscle width, thereby greatly improving the appearance quality of sea cucumbers (especially sea cucumbers).

Description

Technical Field

[0001] The invention belongs to the field of microorganisms and relates to a microbial composition, a microecological preparation and application thereof in breeding sea cucumbers. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] The body wall of sea cucumber is its main edible part and an important part rich in various nutrients. It contains polysaccharides, saponins, collagen, cerebrosides, gangliosides and other bioactive substances. Further studies have shown that it has the effects of enhancing immunity, improving memory, delaying gonadal aging, anti-coagulation, anti-fatigue, anti-tumor, and preventing atherosclerosis and diabetes. The increasing demand for sea cucumber food has led to the booming development of sea cucumber farming. The quality of sea cucumbers, especially the appearance quality, determines the price of sea cucumbers. The body wall and muscle are the main edible parts of sea cucumbers, and are also the main aspects for consumers to evaluate the quality of sea cucumbers. However, the current research and reports on the improvement of sea cucumber quality are still very limited.

[0004] Probiotics refer to live microbial preparations that are beneficial to the host by improving the balance of gastrointestinal microorganisms in animals. Currently, probiotics have been widely used in aquaculture and have the functions of stimulating the development of the host's gastrointestinal tract, improving digestive function, accelerating animal growth, and improving animal immune response and disease resistance. In the breeding process of sea cucumbers, it has been proven that the growth, immunity and disease resistance of sea cucumbers can be improved by adding probiotics to feed or breeding water environment, but there are no reports on the research of improving the quality of sea cucumbers by using probiotics. Summary of the invention

[0005] In order to address the deficiencies of the prior art, the purpose of the present invention is to provide a microbial composition, a microecological preparation and their use in raising sea cucumbers. Raising sea cucumbers with the microbial composition provided by the present invention can greatly improve the appearance quality of sea cucumbers (especially sea cucumbers).

[0006] In order to achieve the above object, the technical solution provided by the present invention is as follows:

[0007] In a first aspect, a microbial composition comprises Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris, wherein the ratio of the number of live bacteria of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1: (0.1~10): (0.1~10).

[0008] The Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris described in the present invention are all microorganisms with biological activity.

[0009] Preferably, the ratio of the viable counts of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1:(0.4~2.5):(0.4~2.5).

[0010] Furthermore, preferably, the ratio of the number of live bacteria of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1:(1.9~2.1):(0.9~1.1).

[0011] In some embodiments, in the microbial composition of the present invention, Pediococcus pentosaceus ( Pediococcus pentosaceus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 11119.

[0012] In some embodiments, in the microbial composition of the present invention, Streptococcus thermophilus ( Streptococcus thermophilus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 10213.

[0013] In some embodiments, in the microbial composition of the present invention, Rhodopseudomonas palustris ( Rhodopseudomonas palustris ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC10649.

[0014] In some embodiments, the present invention feeds the control with the basic feed, and the sea cucumbers fed with the basic feed containing the microbial composition of the present invention and other microorganisms or other microbial compositions are tested for weight and length of each group of sea cucumbers for 56 days, and the fatness is obtained according to the weight and length data. The results show that the fatness of the sea cucumbers fed with the basic feed containing the microbial composition of the present invention is higher, indicating that the microbial composition provided by the present invention can effectively improve the fatness of the sea cucumbers.

[0015] In some embodiments, the present invention uses the basic feed as a control, and the body weight of each group of sea cucumbers fed with the basic feed containing the microbial composition of the present invention and other microorganisms or other microbial compositions is detected after 56 days of body weight and the tare weight of the body wall after absorbing water, and the skin emergence rate is obtained according to the body weight and tare weight data. The results show that the skin emergence rate of sea cucumbers fed with the basic feed containing the microbial composition of the present invention is higher, indicating that the microbial composition provided by the present invention can effectively improve the skin emergence rate of sea cucumbers.

[0016] In some embodiments, the present invention uses the basic feed as a control, and detects the body wall thickness of each group of sea cucumbers fed with the basic feed containing the microbial composition of the present invention and other microorganisms or other microbial compositions on day 56. The results show that the body wall thickness of the sea cucumbers fed with the basic feed containing the microbial composition of the present invention is thicker, indicating that the microbial composition provided by the present invention can effectively increase the body wall thickness of sea cucumbers.

[0017] In some embodiments, the present invention uses the basic feed as a control, and the tubercle height of each group of sea cucumbers fed with the basic feed containing the microbial composition of the present invention and other microorganisms or other microbial compositions is detected on the 56th day. The results show that the tubercle height of the sea cucumbers fed with the basic feed containing the microbial composition of the present invention is higher, indicating that the microbial composition provided by the present invention can effectively increase the tubercle height of sea cucumbers.

[0018] In some embodiments, the present invention uses the basic feed as a control, and the longitudinal muscle width of each group of sea cucumbers fed with the basic feed containing the microbial composition of the present invention and other microorganisms or other microbial compositions is detected on the 56th day. The results show that the longitudinal muscle width of the sea cucumbers fed with the basic feed containing the microbial composition of the present invention is wider, indicating that the microbial composition provided by the present invention can effectively increase the longitudinal muscle width of sea cucumbers.

[0019] In summary, the microbial composition provided by the present invention can effectively improve the appearance quality of sea cucumbers. Therefore, the second aspect of the present invention provides an application of the microbial composition described in the first aspect of the present invention in breeding sea cucumbers.

[0020] Furthermore, breeding sea cucumbers is used to improve the appearance quality of sea cucumbers.

[0021] Furthermore, the sea cucumber is Stichopus japonicus.

[0022] The microbial composition of the present invention can be made into a feed additive or veterinary medicine for use alone, or can be mixed with feed and fed directly.

[0023] In the third aspect, a microbial preparation comprises the microbial composition described in the first aspect of the present invention.

[0024] In some embodiments, the microbial preparation may be in a solid state or in a liquid state.

[0025] Specifically, the solid microbial preparation is a mixture of bacterial powder of the microbial composition and auxiliary materials (such as fillers, etc.) to form powder or granules.

[0026] Specifically, the liquid microbial preparation is a mixture of fermentation broths of each strain of the microbial composition.

[0027] In a fourth aspect, a method for improving the appearance quality of sea cucumbers is provided, comprising mixing the microbial composition described in the first aspect of the present invention with a basic feed to obtain a mixed feed, and then feeding the mixed feed to the sea cucumbers; wherein the total viable count of the microbial composition in the mixed feed is not less than 0.5×10 9 CFU / g.

[0028] In some embodiments, the total viable count of the microbial composition in the mixed feed is (0.5-10)×10 9 CFU / g, preferably (0.5-2)×10 9 CFU / g, more preferably (0.9-1.1)×10 9 CFU / g.

[0029] In some embodiments, the mass of the mixed feed fed is 0.5-2% of the total mass of the sea cucumber.

[0030] In some embodiments, the sea cucumber is Stichopus japonicus.

[0031] The beneficial effects of the present invention are:

[0032] Experiments have shown that feeding a microbial composition provided by the present invention can effectively improve the fatness, skin emergence rate, body wall thickness, papilla height and muscle width of sea cucumbers (especially sea cucumbers), thereby significantly improving the appearance quality of sea cucumbers (sea cucumbers), thereby increasing the market value and economic value of sea cucumbers. DETAILED DESCRIPTION

[0033] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0034] Example 1: Screening of a single probiotic that improves the quality of sea cucumbers

[0035] 1. Experimental methods

[0036] 1.1 Bacteria processing and feed preparation

[0037] The source information of the 10 probiotics studied in this example is shown in Table 1. Activated with MRS liquid medium, streaked on the activated bacterial plate, cultured at 25°C for 48 h, picked a single colony in MRS liquid medium, and cultured at 25°C overnight for growth curve determination. Dilute the bacterial solution to OD600 = 0.399, inoculate 1 mL into 100 mL MRS liquid medium, culture at 25°C and 110 rpm, measure OD600 and spread at 6, 12, 18, 24, 32, 48, 60, and 72 hours, and calculate the number of colonies. The results showed that the concentration of the bacterial solution was 10 at 18h.10 CFU / mL. After centrifugation of the bacterial solution, live bacteria were collected, washed twice with sterile seawater, and an appropriate amount of sterile seawater was added to mix with feed and made into flakes; after centrifugation of the bacterial solution, the supernatant was used as the metabolite of live bacteria for the experiment. First, a part of the supernatant was taken to make dough with feed, and the remaining part was directly added to the culture tank.

[0038] Table 1 Information on the sources of the 10 probiotics used in the experiment

[0039]

[0040] Note: All were purchased from China Agricultural Microbiological Culture Collection Center.

[0041] 1.2 Sea cucumber farming and sample collection

[0042] The experiment was divided into a control group, a Bacillus licheniformis group (BL group), a Bacillus subtilis group (BS group), a Pediococcus acidilactici group (PA group), a Pediococcus pentosaceus group (PP group), a Streptococcus thermophilus group (ST group), an Enterococcus faecalis group (EF group), a Lactobacillus lactis group (LL group), a Lactobacillus plantarum group (LP group), a Rhodopseudomonas palustris group (RP group), and a Candida utilis group (CU group). Each group had three replicates, and 30 sea cucumbers of similar size (25.5 ± 0.4 g, mean ± standard error) were randomly released in each replicate. The control group was fed with a basic feed (commercial formula feed); the experimental group was fed with a feed mixed with the basic feed and the corresponding strains, and the number of live bacteria in the feed of the experimental group was 1×10 9 CFU / g. Add appropriate amount of sterilized seawater, make into flakes, and divide into small pieces for feeding. The feeding amount is 1% of the total body weight of sea cucumbers. Feed at 18:00 every afternoon. Absorb the bottom and replace the seawater before feeding. The temperature, salinity and pH value of seawater are 14-18℃, 30-32‰ and 7.8-8.0 respectively. The experiment lasted for 56 days.

[0043] 1.3 Index detection

[0044] After the experiment, the sea cucumbers were fasted for 24 hours and then weighed (g). A breeding barrel was used as a replicate, and 6 sea cucumbers of uniform size were taken from each barrel. They were placed in a plastic basin in the dark and allowed to stretch. The body length (cm) and the height of the papilla (cm) were measured in water. After the sea cucumbers were dissected and the internal organs were removed, the longitudinal muscle width (cm) was measured with a steel ruler, and the body wall was partially dried to measure the tare weight (g). The longitudinal muscle was then cut off, and the body wall thickness (cm) was measured at 3 points parallel to the back of the body, avoiding the papilla.

[0045] Fullness (g / cm 3 ) = weight / body length 3 .

[0046] Skin removal rate (%) = skin weight / body weight.

[0047] 1.4 Statistical methods

[0048] SPSS 18.0 statistical software was used for data statistics, and one-way ANOVA was used to analyze the treatments. P The difference was considered significant when <0.05.

[0049] 2. Experimental results

[0050] The experimental results are shown in Table 2. The fatness, skinning rate, body wall thickness, papilla height and longitudinal muscle width of the sea cucumbers in the PP, ST and RP groups were significantly higher than those in the control group ( P <0.05, and there were no significant differences in the above indicators between the other treatment groups and the control group ( P> 0.05).

[0051] Table 2 Effects of single probiotic preparation on the quality of sea cucumber

[0052] Note: Each value is mean ± standard error. Different letters in the same row indicate significant differences. P <0.05.

[0053] 3. Experimental conclusion

[0054] From the 10 probiotics selected in this example, three strains were selected that have the effect of improving the quality of sea cucumbers, namely Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris. Whether the above three strains can further improve the quality of sea cucumbers through synergistic effects still needs further research.

[0055] Example 2: Screening of composite microecological preparations for improving the quality of sea cucumbers

[0056] 1. Experimental methods

[0057] 1.1 Bacteria processing and feed preparation

[0058] The strain treatment and feed preparation were the same as in Example 1.

[0059] 1.2 Sea cucumber farming and sample collection

[0060] The experiment was divided into a control group, a Pediococcus pentosaceus group (PP group), a Streptococcus thermophilus group (ST group), a Rhodopseudomonas palustris group (RP group), a combination of Pediococcus pentosaceus and Streptococcus thermophilus group (live bacteria ratio 1:1) (PP-ST group), a combination of Pediococcus pentosaceus and Rhodopseudomonas palustris group (live bacteria ratio 1:1) (PP-RP group), a combination of Streptococcus thermophilus and Rhodopseudomonas palustris group (live bacteria ratio 1:1) (ST-RP group), and a combination of the three bacteria (live bacteria ratio 1:1:1) (PP-ST-RP group). Each group had three replicates, and 30 sea cucumbers of similar size (28.1 ± 0.3 g, mean ± standard error) were randomly released in each replicate. The control group was fed a basic feed (commercial formula feed), and the experimental groups were fed a feed mixed with 1×10 9 CFU / g of live bacteria, the amount of bacteria in the composite group was added according to the proportion of live bacteria, and the total amount of bacteria in the single group was kept consistent. Add an appropriate amount of sterilized seawater, make it into sheets, and divide it into small pieces for feeding. The feeding amount is 1% of the total body weight of the sea cucumber. Feed at 18:00 every afternoon. Absorb the bottom and replace the seawater before feeding. The temperature, salinity and pH value of the seawater are 14-18℃, 30-32‰ and 7.8-8.0 respectively. The experiment lasted for 56 days.

[0061] 1.3 Index detection

[0062] The index detection method is the same as that in Example 1.

[0063] 1.4 Statistical methods

[0064] The statistical method is the same as in Example 1.

[0065] 2. Experimental results

[0066] As can be seen from Table 3, the PP-ST-RP treatment group had the highest fatness, skinning rate, body wall thickness, papilla height and longitudinal muscle width compared with other treatment groups. And the fatness index was significantly higher than that of the PP group ( P <0.05, and the skin peeling rate was significantly higher than that of the PP group, RP group and PP-RP group ( P <0.05, and the body wall thickness was significantly higher than that of the PP group, ST group, PP-ST group and PP-RP group ( P <0.05), and the height of the papilla was significantly higher than that of the PP group, ST group, RP group, PP-RP group and ST-RP group ( P <0.05), and the longitudinal muscle width was significantly higher than that of the PP group, RP group, and ST-RP group ( P <0.05).

[0067] Table 3 Effects of compound microecological preparations on the quality of sea cucumbers

[0068] Note: Each value is mean ± standard error. Different letters in the same row indicate significant differences. P <0.05.

[0069] 3. Experimental conclusion

[0070] After Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris were mixed in pairs and in a ratio of 1:1:1 in terms of viable bacteria count, it was found that the combined addition group had a better effect on improving the quality of sea cucumbers than the single addition group, among which the combination of the three strains in a ratio of 1:1:1 in terms of viable bacteria count had the best effect. Whether the above three strains can further improve the quality of sea cucumbers by optimizing the matching ratio requires further research.

[0071] Example 3 Optimization of the compatibility ratio of the composite microecological preparation for improving the quality of sea cucumbers

[0072] 1. Experimental methods

[0073] 1.1 Bacteria processing and feed preparation

[0074] The strain treatment and feed preparation were the same as in Example 1.

[0075] 1.2 Sea cucumber farming and sample collection

[0076] The experiment set up a control group, a composite group of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris, and the mixture was mixed according to the ratio of viable bacteria count of 1:1:1 (PP-ST-RP group), 1:2:1 (PP-2ST-RP group), 1:1:2 (PP-ST-2RP group), and 2:1:1 (2PP-ST-RP group). Each group had 3 replicates, and 30 sea cucumbers of similar size (35.5± 0.4 g, mean ± standard error) were randomly released in each replicate. The control group was fed with a basic feed (commercial formula feed), and the experimental groups were fed with a feed mixed with 1×10 9 CFU / g of live bacteria, the bacterial amount of the composite group was added according to the proportion of live bacteria, and the bacterial amount of each treatment group was kept consistent. Add an appropriate amount of sterilized seawater, make it into sheets, and divide it into small pieces for feeding. The feeding amount is 1% of the total body weight of the sea cucumber. Feed at 18:00 every afternoon. Absorb the bottom and replace the seawater before feeding. The temperature, salinity and pH value of the seawater are 14-18℃, 30-32‰ and 7.8-8.0 respectively. The experiment lasted for 56 days.

[0077] 1.3 Index detection

[0078] The index detection method is the same as that in Example 1.

[0079] 1.4 Statistical methods

[0080] The statistical method is the same as in Example 1.

[0081] 2. Experimental results

[0082] As shown in Table 4, the fatness, skin emergence rate, body wall thickness, papilla height and longitudinal muscle width of the PP-2ST-RP group were significantly higher than those of the PP-ST-RP group, the PP-ST-2RP group and the 2PP-ST-RP group ( P <0.05).

[0083] Table 4 Effects of different proportions of compound microecological preparations on the quality of sea cucumbers

[0084] Note: Each value is mean ± standard error. Different letters in the same row indicate significant differences. P <0.05.

[0085] 3. Experimental conclusion

[0086] After mixing Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris in the ratio of viable bacteria of 1:1:1, 1:2:1, 1:1:2 and 2:1:1 respectively, it was found that the composite addition group with the ratio of viable bacteria of 1:2:1 had a better effect in improving the quality of sea cucumbers, while the other treatment groups had no further improvement effect.

[0087] In summary, Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris are microecological preparations that can improve the quality of sea cucumbers. At appropriate proportions, the combined addition of the three preparations has a better effect than single addition. Among them, the combined addition of the three preparations at a ratio of 1:2:1 in terms of live bacteria count has the best effect in improving the quality of sea cucumbers.

[0088] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of a microbial composition in raising sea cucumbers, characterized in that: It is used to improve the appearance quality of sea cucumbers in breeding; The appearance quality is fatness, skin emergence rate, body wall thickness, tubercle height and muscle width; The microbial composition comprises Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris, wherein the ratio of the number of live bacteria of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1: (0.1-10): (0.1-10); Pediococcus pentosaceus Pediococcus pentosaceus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 11119; Streptococcus thermophilus ( Streptococcus thermophilus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 10213; Rhodopseudomonas palustris ( Rhodopseudomonas palustris ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 10649.

2. The use according to claim 1, characterized in that: The sea cucumber is Apostichopus japonicus.

3. The use according to claim 1, characterized in that: The ratio of the viable counts of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris was 1: (0.4~2.5): (0.4~2.5).

4. The use according to claim 1, characterized in that: The ratio of the viable counts of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris was 1: (1.9~2.1): (0.9~1.1).

5. A method for improving the appearance quality of sea cucumbers, characterized in that: Mixing a microbial composition with a basic feed to obtain a mixed feed, and then feeding the mixed feed to sea cucumbers; The total viable count of the microbial composition in the mixed feed is not less than 0.5×10 9 CFU / g; The microbial composition comprises Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris, wherein the ratio of the number of live bacteria of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris is 1: (0.1-10): (0.1-10); The appearance quality is fatness, skin emergence rate, body wall thickness, tubercle height and muscle width; Pediococcus pentosaceus Pediococcus pentosaceus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 11119; Streptococcus thermophilus ( Streptococcus thermophilus ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 10213; Rhodopseudomonas palustris ( Rhodopseudomonas palustris ) is from China Agricultural Microbiological Culture Collection Center, with the collection number ACCC 10649.

6. The method according to claim 5, characterized in that: The mass of the mixed feed fed is 0.5~2% of the total mass of the sea cucumber.

7. The method according to claim 5, characterized in that: The ratio of the viable counts of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris was 1: (0.4~2.5): (0.4~2.5).

8. The method according to claim 5, characterized in that: The ratio of the viable counts of Pediococcus pentosaceus, Streptococcus thermophilus and Rhodopseudomonas palustris was 1: (1.9~2.1): (0.9~1.1).

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