Method for quantifying quality of wasp larvae and bee pupae

Abstract

The invention relates to the technical field of biological identification, in particular to a method for quantifying the quality of wasp larvae and bee pupae, and mainly aims to solve the problem that the artificial wasp breeding industry lacks objective quality judgment standards. The method comprises the following steps: freezing a wasp honeycomb so as to conveniently take down larvae and bee pupae, weighing, drying, grinding and the like on a sample, measuring the content of aspartic acid and glutamic acid by utilizing an amino acid analyzer, performing comprehensive calculation by combining indexes such as individual weight and the like, and finally determining the content of aspartic acid and glutamic acid according to a plurality of character values. The quality is judged according to the average content and the total weight of glutamic acid and aspartic acid and the excrement residue condition. The method can effectively distinguish quality differences of wasps of different insect states and species, provides a fair basis for market transactions, promotes sustainable development of the wasp breeding industry, and the effectiveness of the method is verified by specific embodiments.

Description

Technical Field

[0001] The invention relates to the technical field of biological identification, in particular to a method for quantifying the quality of wasp larvae and wasp pupae. Background Art

[0002] Artificial wasp farming is a significant aquaculture industry that has developed in recent years. Wasp larvae and pupae are common agricultural products, and fried versions are popular. Artificial wasp farming has greatly increased the supply of larvae and pupae in the market. However, when wasp farmers trade larvae and pupae in the market, they often lack the quality assessment methods and techniques to determine the quality of different larvae and pupae. They are also unable to assess the quality of larvae and pupae from the same wasp species at different times or from different sources. This lack of assessment methods and techniques hinders the fair trade of larvae and pupae and the healthy and sustainable development of the wasp farming industry.

[0003] Since there is currently no method to quantify the quality of hornet larvae and pupae, market transactions have never taken into account the differences in quality of hornet larvae and pupae. The lack of relevant technology is not conducive to the fairness of transactions and undermines the enthusiasm of both hornet breeders and hornet consumers. Summary of the Invention

[0004] Based on this, the object of the present invention is to provide a method for quantifying the quality of wasp larvae and wasp pupae to solve the technical problem.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A method for quantifying the quality of wasp larvae and pupae comprising:

[0007] (1) Place freshly collected hornet combs in a -20°C refrigerator and quickly freeze the larvae and pupae for 8 hours to easily remove them from the combs;

[0008] (2) Remove the larvae and pupae, randomly select 10 of them and weigh them, then immediately place them in a 70°C oven to dry for 12 hours until there is no moisture left and weigh them again to calculate the dry weight;

[0009] (3) Ten dried larvae or pupae were mixed together, ground into powder, and 20 mg of the sample was randomly taken for amino acid determination;

[0010] (4) The sample was hydrolyzed and digested with 6 mol / L concentrated hydrochloric acid at 110°C under nitrogen for 22 h until all proteins were decomposed into amino acids. The aspartic acid and glutamic acid contents in mg / g were then determined using an amino acid analyzer.

[0011] (5) Based on the average value of glutamic acid content g4_1 and the average value of aspartic acid content g4_2, the sum of the two is calculated as g5=g4_1+g4_2; then, based on the weight of the individual wasp, the total amount of glutamic acid and aspartic acid in the individual wasp is calculated;

[0012] (6) The quality of wasp larvae and pupae was quantified by integrating the mean values ​​of glutamate and aspartate g4_1 and g4_2 in each individual, the sum of the mean values ​​of glutamate and aspartate in each individual, the total weight of glutamate and aspartate in each individual, and the presence of larval feces in each individual.

[0013] (7) The quality of wasps can be determined comprehensively based on the trait values, without relying on a single trait value:

[0014] 1. Average glutamate and aspartate content in individuals (g4);

[0015] 2. The sum of the average content of glutamate and aspartate in an individual (g5);

[0016] 3. The total weight of glutamate and aspartate in the individual;

[0017] 4. Whether there are residual larval feces in the individual;

[0018] 5. The higher the calculated glutamate and aspartic acid values, the better the quality and the more popular it is with consumers. Glutamate and aspartic acid content can be used to evaluate the quality of different wasp stages and the quality of different wasps within the same stage.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 1) The quality of wasps has long received little attention. Through preliminary experiments, we have developed a method to quantify wasp quality based on glutamate and aspartic acid content. This method could potentially allow researchers to quantify or evaluate the quality of wasp larvae and pupae based on these two amino acids, particularly glutamate.

[0021] 2) Quantifying the quality of wasp larvae and pupae using specific amino acid content provides wasp breeders, operators, and consumers with more specific information about their quality. This not only improves transaction fairness but also provides a theoretical basis for resolving disputes in market transactions. DETAILED DESCRIPTION

[0022] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0023] The following describes an embodiment of the present invention based on its overall structure.

[0024] Artificial wasp farming is a significant aquaculture activity that has developed in recent years. Wasp larvae and pupae are common agricultural products, and fried versions are popular. Artificial wasp farming has greatly increased the supply of larvae and pupae in the market. However, when wasp farmers trade these agricultural products, they often lack the basis and technology to determine the quality of different larvae and pupae. They are also unable to determine the quality of larvae and pupae from the same wasp species at different time periods or from different geographic subspecies. This lack of quality assessment methods and techniques hinders the fair trade of larvae and pupae products and the healthy and sustainable development of the wasp farming industry. Therefore, there is an urgent need to explore methods for determining the quality of agricultural products such as larvae and pupae. This patent application proposes a method and technology for quantifying the quality of larvae and pupae, providing technical support for sustainable wasp farming.

[0025] Currently, there are no existing methods for evaluating agricultural products such as wasp larvae and bee pupae. Their quality is not considered during the trading process. While consumers can perceive the differences between fried wasp products, such as the bicolor wasp having a better taste than the golden-ringed wasp, there is no specific method to quantify this. This application proposes a method for quantifying the quality of wasp larvae and bee pupae.

[0026] After searching relevant databases, we did not find any patents that provide technologies and methods for quantifying the quality of wasp larvae and pupae. Based on this, we proposed a method to quantify the quality of wasp larvae and pupae based on the amino acid and aspartic acid content in them.

[0027] A search of existing patent databases revealed a patent application for quick freezing of wasp larvae and bee pupae (application number: CN202410946930.X). This patent application proposes a method for freezing wasp larvae and bee pupae. The first step also requires quick freezing of wasp larvae and bee pupae. However, in terms of content, this application is completely different from the patent (CN202410946930.X).

[0028] In daily life, people often use glutamate to evaluate food quality, but no one has used glutamate content to evaluate the quality of wasp larvae and bee pupae. In academia, glutamate is often used to evaluate food freshness (e.g., Zhang Songhong, 2008. Freshness Evaluation of Monosodium Glutamate. Chinese Condiments, 6, 31-33; Gu Yanjun, 2012. On the Quantification of Umami and Its Application in Compound Seasonings. Food Industry, 33, 141-143; Xue Song, 2010. Effects of Ice Storage on the Freshness and Free Amino Acids of Chicken. Jiangsu Agricultural Sciences, 6, 411-413). However, existing research has not established a relationship between glutamate content and the quality of wasp pupae and larvae. Through extensive preliminary research, we have found that glutamate and aspartic acid are correlated with the quality of wasp larvae and bee pupae. This serves as a theoretical basis for this patent. Based on this, we propose a method for quantifying the quality of wasp larvae and bee pupae using glutamate and aspartic acid.

[0029] Currently, there is no method for quantifying the quality of wasp larvae and pupae. This patented technology, based on our own measured data and combined with existing research, proposes, for the first time, a simple method for quantifying the quality of wasp larvae and pupae. This method allows wasp breeders and market traders to accurately and conveniently evaluate wasp quality and can also provide insights for researchers conducting research on wasp pupae.

[0030] 1. Freezing of honeycomb

[0031] Place freshly collected hornet combs in a -20°C freezer and rapidly freeze the larvae and pupae for at least 8 hours. Remove the larvae and pupae from the frozen combs. Separate the larvae and pupae based on the test subject's needs. Immediately proceed with the next step of processing the collected material.

[0032] 2. Determination and calculation of fresh wasp weight

[0033] From the larvae and pupae taken out, 10 larvae or pupae were randomly selected as one sample, and 5 replicates were set up (thus a total of 50 samples were analyzed).

[0034] After weighing (g1) each sample, immediately place it in a 70°C oven and continue drying for 12 hours until there is no moisture and weigh it again (g2). The dry weight is calculated as g3 = g1-g2.

[0035] 3. Grind the sample into powder

[0036] Ten larvae (or pupae) from each sample were mixed and thoroughly ground into a powder in a clean container. 20 mg of the powder was randomly sampled for amino acid determination.

[0037] 4. Amino acid determination

[0038] Determine the amino acid content of powdered samples according to the national standard GB / T5009.124-2016. Hydrolyze and digest the sample with concentrated hydrochloric acid (6 mol / L) at 110°C under nitrogen for 22 hours until all proteins are broken down into amino acids. Determine the aspartic acid and glutamic acid content (mg / g) using an amino acid analyzer (e.g., Biochrom 30+).

[0039] 5. Calculation of total glutamate and aspartic acid in samples

[0040] Based on the average value of glutamic acid and / or aspartic acid content (g 4_1 and g 4_2 ), calculate the sum of the two (g5=g 4_1 +g 4_2 ); then calculate the total amount of glutamate and aspartic acid in the individual wasp based on its individual weight (g6=g3*g5).

[0041] 6. Quality evaluation of wasp pupae

[0042] In order to accurately quantify the quality of wasp larvae and pupae, the following traits are integrated for comprehensive judgment rather than relying on a single trait value:

[0043] A. Mean values ​​of glutamate and aspartate in individuals g 4-1 and g 4-2 ;

[0044] B, the sum of the mean values ​​of glutamate and aspartate in an individual g5;

[0045] C, the total weight of glutamate and aspartate in the individual g6;

[0046] D. Whether there are residual larval feces in the individual.

[0047] The following is a detailed description of a specific case:

[0048] 1. Quality Assessment of Vespa velutina Larvae and Pupae

[0049] In production practice, the yellow-legged vespa is the most commonly cultivated species. Four treatments were selected, including young larvae, old larvae, young pupae, and old pupae of the yellow-legged vespa. In this example, three replicates were set for each treatment, and 10 individuals were selected from each treatment.

[0050] The contents of 17 common amino acids (aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, arginine, and proline) were determined using the same method as above. The results are shown in Table 1.

[0051] Table 1. Contents of 17 common amino acids in young larvae, old larvae, young pupae, and old pupae of yellow-legged wasps*

[0052]

[0053] *The maximum values ​​of glutamate and aspartate contents are shown in bold.

[0054] To judge the quality of yellow-legged wasp larvae and pupae, we use the following traits to make a comprehensive judgment, rather than relying on a single trait:

[0055] 1. Mean values ​​of glutamate and aspartate in individuals g 4_1 and g 4_2 ;

[0056] 2. The sum of the mean values ​​of glutamate and aspartate in an individual, g5;

[0057] 3. The total weight of glutamate and aspartate in the individual;

[0058] 4. Whether there are any residual larvae and feces in the individual.

[0059] The quantitative indicators are shown in Table 2.

[0060] Table 2 Calculated values ​​of quality-related traits of young larvae, old larvae, young pupae and old pupae of Vespa lutea

[0061]

[0062] *Estimated based on half the value of yellow-legged wasp feces; ※ Higher value.

[0063] Based on the above four criteria, as calculated in Table 2, the quality of younger pupae and older larvae of the yellow-legged wasp is higher than that of younger larvae and older pupae. This quantitative result is consistent with the common understanding that younger pupae are more popular in the market. The patent application provides a specific quantitative method.

[0064] 2. Quality Assessment of Vespa basalis Larvae and Pupae

[0065] Four treatments were selected for the young larvae, the old larvae, the young pupae, and the old pupae of the basal wasp. Three replicates were set for each treatment, with 10 individuals selected from each. The contents of 17 common amino acids (aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, arginine, and proline) were measured. The assay methods were as described above, and the results are shown in Table 3.

[0066] Table 3 Contents of 17 common amino acids in young larvae, old larvae, young pupae and old pupae of the basal wasp*

[0067]

[0068]

[0069] *The maximum values ​​of glutamate and aspartate contents are shown in bold.

[0070] We quantified the quality of larvae and pupae of the basic wasp using the above indicators. The results of the quantitative indicators are shown in Table 4.

[0071] Table 4. Calculated values ​​of quality-related traits of young larvae, old larvae, young pupae, and old pupae of Vespa basilis

[0072]

[0073] According to the weight ratio, it is estimated to be 1.5 times that of the yellow-legged wasp;

[0074] *Estimated based on half the value of basal wasp feces;

[0075] ※ Higher value.

[0076] According to the above four criteria, as calculated from Table 4, the quality of young pupae of Vespa biocolor is higher than that of old larvae, young larvae, and old pupae. The results of the patent application are consistent with the common understanding that young pupae are more popular in the market. The patent application provides a specific quantitative method. III. Quality determination of larvae and pupae of Vespa biocolor

[0077] Four treatments were selected for the bicolor wasp: young larvae, old larvae, young pupae, and old pupae. Three replicates were set for each treatment, with 10 individuals selected from each. The contents of 17 common amino acids (aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, arginine, and proline) were measured. The assay method is as described above, and the results are shown in Table 5.

[0078] Table 5 Contents of 17 common amino acids in young larvae, old larvae, young pupae and old pupae of Wasp bicolor*

[0079]

[0080]

[0081] *The maximum values ​​of glutamate and aspartate contents are shown in bold.

[0082] We quantified the above indicators for the quality of bicolor hornet larvae and pupae. The results of the quantitative indicators are shown in Table 6.

[0083] Table 6 Calculated values ​​of quality-related traits of young larvae, old larvae, young pupae and old pupae of Wasp bicolor

[0084]

[0085] Based on the body weight ratio, the estimate is 1.0 times that of the yellow-legged vespa. *Estimated based on half the value of the vespa feces. ※ Higher value.

[0086] Based on the above four criteria, as calculated in Table 6, the quality of younger pupae of the bicolor wasp is higher than that of older larvae, younger larvae, and older pupae. This quantitative result is consistent with the common understanding that younger pupae are more popular in the market. This provides a specific quantitative method.

[0087] 4. Quality determination of larvae and pupae of different species

[0088] Based on the previously quantified results of the quality of the yellow-legged wasp, basic wasp and two-colored wasp, the young pupae of the three types of wasps were placed together for comparative analysis to see which wasp had better quality.

[0089] Table 7 Quality evaluation of different wasp larvae and pupae

[0090]

[0091] ※ Higher value.

[0092] Based on the above four criteria, as calculated from Table 7, the glutamic acid content and aspartic acid content per unit weight of young pupae of Vespa basilis are the highest, as well as the total weight of glutamic acid and aspartic acid per individual. Therefore, it can be concluded that the quality of young pupae of Vespa basilis is higher than that of Vespa fusca and Vespa bicolor. Specific quantification method.

[0093] The above-described embodiments are only used to illustrate the technical solutions of the present application, rather than to limit them. Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. These modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present application, and should all be included in the scope of protection of the present application.

Claims

1. A method for quantifying the quality of wasp larvae and wasp pupae, characterized in that: include: (1) Place freshly collected hornet combs in a -20°C refrigerator and quickly freeze the larvae and pupae for 8 hours to easily remove them from the combs; (2) Remove the larvae and pupae, randomly select 10 of them and weigh them g1, immediately place them in a 70°C oven and continue drying for 12 hours until there is no moisture, and weigh them again g2, from which the dry weight g3 is calculated as g1-g2; (3) Ten dried larvae or pupae were mixed together, ground into powder, and 20 mg of the sample was randomly taken for amino acid determination; (4) The sample was hydrolyzed and digested with 6 mol / L concentrated hydrochloric acid at 110°C under nitrogen for 22 h until all proteins were decomposed into amino acids. The aspartic acid and glutamic acid contents in mg / g were then determined using an amino acid analyzer. (5) Based on the average value of glutamic acid content g 4_1 and the average value of aspartic acid content g 4_2 , calculate the sum of the two g5 = g 4_1 +g 4_2 ; Then, based on the weight of the individual wasp, calculate the total amount of glutamic acid and aspartic acid in the individual g6 = g3 * g5; (6) The mean values ​​of glutamate and aspartate in the integrated individual g 4_1 and g 4_2 The quality of wasp larvae and pupae was quantified by the following characteristics: the sum of the average values ​​of glutamate and aspartic acid in individuals g5, the total weight of glutamate and aspartic acid in individuals g6, and whether there was residual larval feces in the individuals. (7) The quality of wasps can be determined comprehensively based on the trait values, without relying on a single trait value:

1. Average glutamate and aspartate content in individuals (g4); 2. The sum of the average content of glutamate and aspartate in an individual (g5); 3. The total weight of glutamate and aspartate in the individual; 4. Whether there are residual larval feces in the individual; 5. The higher the calculated glutamate and aspartic acid values, the better the quality and the more popular it is with consumers. Glutamate and aspartic acid content can be used to evaluate the quality of different wasp stages and the quality of different wasps within the same stage.

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