A feed for adult male chinese pangolin

Abstract

The application discloses adult male Chinese pangolin feeding food, according to the mass percentage of dry matter, the total chitin content in the feeding food is 5.5%-6.5%, which comprises bee pupa 9.30%, silkworm pupa dry 6.47%, Polyrhachis vicina 30.00%, black soldier fly larvae dry 21.30%, breadworm dry 10.64%, cricket dry 1.30%, earthworm dry 1.70%, housefly larvae 5.30%, soil 11.92%, chitin 2.07%. The total chitin content in the preferred formula is 6.00%. The application sets five chitin gradient levels of 2%-10%, and systematically investigates the multi-dimensional influence on the growth performance, blood biochemistry, antioxidant immunity, nutrient digestion rate and intestinal flora of adult male Chinese pangolin, and for the first time determines that the optimal chitin content in the feeding food is 6%. The feeding food can meet the daily nutritional needs of adult male Chinese pangolin, realizes stable weight gain, optimizes intestinal health and immune function, and provides technical support for artificial feeding and ex-situ protection of Chinese pangolin.

Description

Technical Field

[0001] This invention belongs to the field of animal nutrition and feed science, and specifically relates to a diet for adult male Chinese pangolins. Background Technology

[0002] The Chinese pangolin (Manis pentadactyla) belongs to the order Pholidota, family Manidae, and genus Manis. In 2019, it was listed as Critically Endangered (CR) on the IUCN Red List of Threatened Species, and in 2020, my country upgraded its national key protection level to Level I. With the continued decline in wild populations, artificial rescue and captive breeding have become core intervention methods for protecting this species. In the wild, the Chinese pangolin exclusively feeds on termites and ants. These natural food sources provide not only conventional nutrients such as protein and fat, but also a large amount of chitin. Chitin, also known as chitosan, is composed of N-acetyl-D-glucosamine and D-glucosamine linked by a β-1,4 glycosidic bond (Dai et al., 2020) and is widely found in the exoskeletons of crustaceans, insect exoskeletons, and fungal cell walls (Satitsriet et al., 2020). Existing studies have confirmed that chitin and its derivative chitosan have multiple physiological functions, including regulating immunity, anti-oxidation, and improving gut health. It is considered the seventh core nutrient element after protein, fat, carbohydrates, vitamins, minerals, and water (Liu Yuepeng et al., 2024). In animal nutrition, adding 0.1% chitin fermentation broth to the feed of grass carp and silver carp juveniles can significantly improve growth performance (Wang Juhua et al., 2016); adding 3% chitin to the basal diet of broiler chickens can improve weight gain and survival rate while reducing feeding costs (Wang Lixin et al., 2003); adding 1.0 g / kg chitin to the feed of Nile tilapia can simultaneously promote growth and enhance immunity and disease resistance (Kumaran et al., 2021).

[0003] However, the formulation of artificial diets for Chinese pangolins in captivity faces unique challenges. On the one hand, the supply of natural ants is difficult to maintain consistently, and existing artificial feed formulations often use conventional insect raw materials as substitutes, resulting in significant differences in chitin content compared to natural food sources. On the other hand, research on chitin nutrition in pangolins is still not systematic. Lin et al. (2015) reported that adding 5% chitin to the feed of Chinese pangolins can improve fecal characteristics and reduce the digestibility of dry matter, protein, and energy, which has a positive effect on obesity control in captive individuals. Cabana et al. (2019) found in a feeding experiment of Malayan pangolins (Manis javanica) that when the chitin content in the diet was 10%, it could assist in the digestion and absorption of organic matter, crude protein, and fiber without affecting the palatability of the feed. The above studies provide a preliminary basis for the application of chitin in pangolin feed, but there are still the following shortcomings: (1) The research subjects involve different species (Chinese pangolin and Malayan pangolin), and the differences in digestive physiology between species have not been clarified; (2) Existing studies have only examined single or limited chitin levels, and have not systematically compared the comprehensive effects of different addition amounts on pangolins; (3) Most importantly, the appropriate amount of chitin required by Chinese pangolins has not been determined to date, and whether additional chitin should be added to artificial feed, and how much should be added to balance growth performance, digestive health and immune function, are all lacking systematic experimental data support.

[0004] This technological gap directly restricts the development of artificial compound feeds that meet the nutritional needs of Chinese pangolins and have appropriate chitin levels, becoming a major technical bottleneck in the current work of artificial rescue and captive breeding of pangolins.

[0005] In view of this, this invention systematically investigated the multidimensional effects of chitin on the growth performance, blood biochemistry, antioxidant immunity, nutrient digestibility and intestinal flora of adult Chinese pangolins by setting gradient chitin levels (2%~10%). For the first time, the optimal chitin content in their diet was quantitatively determined to be 6%, providing direct experimental evidence and formulation scheme for overcoming the above-mentioned technical bottlenecks. Summary of the Invention

[0006] To improve the nutritional requirements of Chinese pangolins and explore systematic research on chitin nutrition, this invention provides a chitin-optimized diet for adult male Chinese pangolins, thereby improving the survival quality of pangolins in current artificial rescue and captive breeding programs.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] This invention first discloses a diet for adult male Chinese pangolins, comprising the following ingredients by weight percentage:

[0009] Bee pupae 9.30%, dried silkworm pupae 6.47%, black spiny ant 30.00%, dried black soldier fly larvae 21.30%, dried mealworm 10.64%, dried cricket 1.30%, dried earthworm 1.70%, housefly larvae 5.30%, soil 11.92%, chitin 2.07%.

[0010] Furthermore, the total chitin content in the diet is 5.5% to 6.5%.

[0011] Furthermore, the total chitin content in the diet is 6.0%.

[0012] The present invention also discloses a feeding method for adult male Chinese pangolins according to any of the above-mentioned feeds, comprising: crushing and mixing adult male Chinese pangolin feed and water at a mass ratio of 1:1.8 and feeding the adult male Chinese pangolins once a day, allowing them to eat freely.

[0013] The beneficial effects of this invention are as follows:

[0014] 1. The optimal total chitin level in the diet of adult Chinese pangolins was determined for the first time. By setting five chitin gradient levels (2%, 4%, 6%, 8%, and 10%), the effects of different chitin contents on 15 core physiological indicators of adult male Chinese pangolins were systematically investigated. The results showed that the dietary chitin level and the overall health score exhibited a typical unimodal nonlinear relationship (fitting equation: y = -0.030161χ). 2 + 0.389949χ - 0.456240, R 2 = 0.9993), the theoretical optimal value is 6.46%, and the experimental verification shows that the optimal value is 6%.

[0015] 2. This invention is the first to discover that the effect of chitin levels on pangolin health is not a simple linear relationship: when the chitin content is below 5.5%, the body's antioxidant capacity is insufficient and the number of beneficial intestinal bacteria decreases; when it is above 6.5%, the digestibility of crude fat decreases significantly, catalase activity is inhibited, and the number of opportunistic pathogens in the intestine increases; only within the range of 5.5% to 6.5% does growth performance, digestive function, immune defense, and intestinal microecology simultaneously reach an optimal balance. This discovery of the "optimal point" cannot be reasonably predicted by those skilled in the art based on existing technology.

[0016] 3. Comprehensive improvement of multiple physiological indicators. Using the diet of this invention with 6% total chitin, adult Chinese pangolins can achieve stable weight gain, with significantly better daily feed intake and apparent digestibility of crude fat than other levels. Feces have good shape and appropriate moisture content, serum antioxidant capacity and lysozyme activity are optimal, beneficial bacteria in the intestine are enriched and harmful bacteria are reduced, and core metabolic pathways such as lipids and amino acids are normal. Detailed Implementation

[0017] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0018] The following examples were conducted at the Pangolin Conservation and Research Center of the National Forestry and Grassland Administration (hereinafter referred to as the "Pangolin Conservation and Research Center"). Since 2020, the center has been engaged in the rescue and breeding of pangolins, achieving a rescue success rate of 80%, with many Chinese pangolins successfully becoming pregnant and giving birth to cubs. Unless otherwise specified, the methods used in the following examples are conventional methods; and unless otherwise specified, the raw materials used are commercially available products.

[0019] Example 1

[0020] Experiment to determine the optimal level of total chitin in the diet of adult male Chinese pangolins

[0021] 1. Materials and Methods

[0022] 1.1 Experimental subjects and experimental design

[0023] The Chinese pangolin is a critically endangered species with a small captive population. This study used the Pangolin Conservation and Research Center of the National Forestry and Grassland Administration as its experimental base, which houses the largest captive population of Chinese pangolins in China. Ten healthy male adult pangolins of similar weight were selected as subjects. Five dietary chitin levels were established: 2% (Group A), 4% (Group B), 6% (Group C), 8% (Group D), and 10% (Group E). The experiment was conducted from September to December 2025, consisting of five 14-day phases, including a 7-day pre-trial period and a 7-day main trial period.

[0024] A 7-day washout period was set between each stage, during which the basal diet was fed to eliminate the residual effect of the previous stage treatment.

[0025] 1.2 Experimental diets and feeding management

[0026] Table 1 shows the composition and measured nutrient levels of the experimental diets in their air-dried state (i.e., dry matter) as percentages by mass. In accordance with pangolin husbandry and management regulations, all experimental animals were housed individually during the experiment, with daily cleaning and regular disinfection. Feeding was conducted once daily at 16:00, with each feeding amount being 160 g (diet ingredients:water = 1:1.8), allowing free access to food and water. Actual feed intake was recorded at 09:00 the following day. Body condition monitoring was performed on the experimental animals during the experiment, and fasting body weight was measured daily.

[0027] Feed preparation method: Weigh the prepared feed ingredients according to the feed composition ratio in Table 1, add the weighed feed ingredients and water in a mass ratio of 1:1.8 into a blender and crush and mix evenly.

[0028] Table 1. Dietary composition and measured nutrient levels (air-dried basal, %)

[0029]

[0030] Note: The "Chitin" column in Table 1 represents the additional amount added; after measuring and converting the background chitin content of the raw materials, the total chitin content of the diets in each experimental group was 2%, 4%, 6%, 8%, and 10%, respectively. The proportion of chitin-containing raw materials such as bee pupae and dried silkworm pupae was consistent in each group, and the total chitin level was controlled only by adjusting the proportion of sterilized soil and additional chitin added.

[0031] 1.3 Sample Collection and Index Testing

[0032] 1.3.1 Stool score and water content

[0033] Pangolin feces were observed and recorded daily, and a fecal score was assigned. The fecal scoring criteria were developed based on the methods of Grellet (2012), Clark (2016), etc., as shown in Table 2. Fecal moisture content was determined using the conventional drying method, and the mass (W) of fresh fecal samples was accurately weighed. 鲜粪 The product was dried in an oven at 65°C until constant weight, and then weighed after cooling (W). 绝干粪 The water content of feces is calculated according to formula (1).

[0034] Table 2 Stool Scoring Criteria

[0035]

[0036] (1)

[0037] 1.3.2 Apparent digestibility of nutrients

[0038] Feed samples were collected during the trial period, and feces were collected continuously for 7 days using the full collection method. After weighing and recording, the feces were dried at 60℃ and stored for later testing.

[0039] Dry matter (DM) in feed and feces was determined according to "Determination of Moisture in Feed" (GB / T 6435-2006); crude ash (Ash) was determined according to "Determination of Crude Ash in Feed" (GB / T 6438-2007); crude protein (CP) was determined according to "Determination of Crude Protein in Feed - Kjeldahl Method" (GB / T 6432-2018); crude fat (EE) was determined according to "Determination of Crude Fat in Feed" (GB / T 6433-2006); calcium (Ca) was determined according to "Determination of Calcium in Feed" (GB / T 6436-2018); total phosphorus (TP) was determined according to "Determination of Total Phosphorus in Feed - Spectrophotometric Method" (GB / T 6437-2018); and total energy (GE) was analyzed using an HXR-6000 fully automatic oxygen bomb calorimeter (Hunan Huaxing Energy Technology Co., Ltd.). The calculation formulas for the relevant indicators are as follows:

[0040] Formula (2)

[0041] DMD%—apparent digestibility of dry matter in fecal samples;

[0042] DM i —Dry matter intake, g;

[0043] DM e —Mass of dry matter excreted in feces, g.

[0044] Formula (3)

[0045] GED% — Apparent energy digestibility in fecal samples;

[0046] GE i —Total energy intake, kJ;

[0047] GE e —Total energy of feces, kJ.

[0048] Formula (4)

[0049] ND% — Apparent digestibility of crude protein, crude fat, crude ash, and other substances in fecal samples;

[0050] N i —Nutrient intake, g;

[0051] N e —Amount of nutrients excreted in feces, g.

[0052] Formula (5)

[0053] C d —Chitin content in the diet, %

[0054] C f —Chitin content in feces, %

[0055] F—fecal dry matter output, g;

[0056] D—Dietary dry matter intake, g.

[0057] 1.3.3 Blood biochemistry, antioxidant and immune indicators

[0058] The prepared plasma was analyzed using a fully automated biochemical analyzer to determine its biochemical parameters. The reagents used were 17 biochemical parameters from IDEXX Corporation (USA), including total protein (TP), albumin (ALB), globulin (GLOB), albumin / globulin ratio (ALB / GLOB), urea (UREA), creatinine (CREA), blood urea nitrogen / creatinine ratio (BUN / CREA), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), total bilirubin (TBIL), glucose (GLU), cholesterol (CHOL), amylase (AMYL), lipase (LIPA), calcium (Ca), and phosphorus (PHOS).

[0059] 1.3.4 Antioxidant Indicators

[0060] Separated serum samples were used for antioxidant and immune marker detection. A total of seven serum antioxidant markers were measured, including: total antioxidant capacity (T-AOC), malondialdehyde (MDA), oxidized glutathione (GSSG), reduced glutathione (GSH) levels, and the activities of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). One immune-related marker, lysozyme (LZM) activity, was also measured. All procedures were strictly performed according to the kit instructions.

[0061] 1.3.5 Gut microbiota analysis

[0062] Fresh fecal samples were collected, total microbial DNA was extracted, the V3–V4 region of the 16S rRNA gene was amplified, and high-throughput sequencing was performed to analyze microbial diversity and species composition.

[0063] 1.3.6 Construction of Comprehensive Evaluation Model and Curve Fitting To determine the optimal chitin addition level, indicators with clear biological significance and significant inter-group differences were selected from growth performance, feeding behavior, fecal characteristics, nutrient digestibility, blood biochemistry, antioxidant activity, immune activity, and gut microbiota structure to construct a multi-dimensional comprehensive evaluation system. All indicators were standardized to the 0-1 range, and the comprehensive score for each group was calculated using an equal-weighted assignment method. With chitin levels (2%, 4%, 6%, 8%, 10%) as the independent variable (χ) and the comprehensive score as the dependent variable (y), a quadratic polynomial model was fitted: y = aχ 2 + bχ + c, passing through the extreme point χ opt = -b / (2a) to solve for the theoretically optimal addition level.

[0064] 1.3.8 Data Statistics and Analysis

[0065] All data were initially processed using Excel, and then statistical analysis was performed using SPSS 26.0 software (IBM, USA). Graphs were generated using R 4.2.3 (R Core Team, 2023). The specific analysis steps and methods are as follows:

[0066] First, all data were tested for normality (Shapiro-Wilk test, P > 0.05) and homogeneity of variance (Lvene's test, P > 0.05). Data that met the parametric test requirements were then used for further analysis. One-way ANOVA was used to analyze data between different feed chitin levels. Duncan's method was used for multiple comparisons when differences were significant. Kruskal-Wallis H test was used for data that did not conform to a normal distribution. Results are expressed as mean ± standard error (Mean ± SEM). P < 0.05 was considered significant, and P < 0.01 was considered highly significant.

[0067] 2 Results

[0068] 2.1 Effects of dietary crude protein levels on body weight and average daily feed intake of adult Chinese pangolins

[0069] Table 3. Effects of dietary crude protein levels on body condition of adult Chinese pangolins

[0070]

[0071] Table 3 shows that there were no significant differences in the initial and final weights of pangolins during each experimental period (P>0.05), but in groups A and E, there was a negative growth trend in body weight. The average daily food intake of each group showed significant differences (P < 0.05), with the average daily food intake of groups A and E being significantly lower than that of groups B, C, and D (P < 0.05).

[0072] In the daily feeding and management of adult pangolins, body weight and feed intake are important monitoring indicators. In this experiment, the total energy of the different crude chitin levels of the designed diets were similar (Table 1). The feeding amount of pangolins during each stage of the trial period (160 g per day, with feed ingredients: water = 1:1.8) was the same, and the average body weight of pangolins remained between 4.47 and 4.89 kg (Table 3). This indicates that the diet set in this experiment can meet the daily needs of pangolins and there is no situation of overnutrition.

[0073] 2.2 Effects of dietary crude protein level on fecal score and moisture content of adult Chinese pangolins

[0074] Table 4. Effects of dietary crude protein level on fecal scores of adult Chinese pangolins

[0075]

[0076] Note: Different lowercase letters in the superscript of the same data indicate significant differences (P < 0.05), while no letter or the same letter indicates no significant differences (P > 0.05). The same applies to the table below.

[0077] Table 4 shows that after feeding with diets of different crude protein levels, the fecal scores of pangolins ranged from 2.58 to 3.58, and the water content ranged from 49.88% to 60.32%. There were highly significant differences in fecal scores and water content among the groups (P < 0.001), and both indicators showed a consistent trend. Group A had the lowest fecal score (2.58±0.08) and the highest moisture content (60.32±1.40%), significantly different from most other groups (P < 0.05). Groups B and C had moderate fecal scores (3.07±0.11 and 3.13±0.06, respectively) and moisture content (56.08±1.66% and 57.53±1.05%, respectively), with no significant difference between the two groups (P > 0.05). Groups D and E had the highest fecal scores (3.51±0.10 and 3.58±0.12, respectively) and the lowest moisture content (52.91±1.02% and 49.88±0.82%, respectively), with no significant difference between the two groups (P > 0.05). Groups B, C, and D were in relatively ideal condition. Therefore, it can be preliminarily concluded that a diet with appropriate chitin levels has no adverse effects on the intestinal health of Chinese pangolins.

[0078] 2.3 Effects of dietary crude protein levels on plasma biochemical parameters of adult Chinese pangolins

[0079] Table 5. Effects of dietary crude protein levels on plasma biochemical parameters of adult Chinese pangolins.

[0080]

[0081]

[0082] Changes in blood biochemical parameters are closely related to the absorption and metabolism of dietary nutrients by animals. Changes in plasma TP, ALB, and BUN can reflect the animal's protein metabolism and nutritional status. Plasma TP, to a certain extent, represents the nutritional level of dietary protein and the degree of protein digestion and absorption by the animal. GLU reflects the body's glucose intake and utilization level; TG and CHOL are the main forms of lipids in the blood. TG participates in the body's substance and energy metabolism, and its content reflects the body's lipid absorption, utilization, and metabolism; LDL-C and HDL-C participate in the metabolic transport of CHOL within and outside the liver. LDH is an important indicator of kidney function; pancreatic damage is the most significant cause of increased LIP levels; ALP is an important indicator of liver damage; ALT is closely related to protein metabolism and liver function and is often used as an indicator for diagnosing liver function.

[0083] As shown in Table 5, there were no significant differences among the groups in terms of chitin content in the diet for conventional biochemical indicators such as blood glucose, creatinine, blood urea nitrogen, calcium ions, total protein, albumin, and globulin, as well as liver function-related indicators such as alanine aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase, metabolic indicators such as total bilirubin and cholesterol, and digestive enzyme indicators such as pancreatic amylase and lipase (P > 0.05). The blood urea nitrogen / creatinine ratio was significantly lower in group C than in groups A, B, and E (P < 0.05), while there was no significant difference between group D and the other groups (P > 0.05). Overall, the trend was C < D < A, B, and E. Phosphate ion content was highest in group E, significantly higher than in groups A, B, C, and D (P < 0.05), while there was no significant difference among groups A, B, C, and D (P > 0.05). The albumin / globulin ratio was significantly lower in group B than in groups C, D, and E (P < 0.05), while there was no significant difference between group A and the other groups (P > 0.05), and no significant difference among groups C, D, and E (P > 0.05). These results indicate that dietary chitin levels have no significant adverse effects on the core physiological metabolism of adult Chinese pangolins, mainly regulating protein metabolism, mineral balance, and the proportion of immune proteins through dose-effect regulation. Appropriate chitin levels can optimize protein metabolism and maintain immune balance, while high chitin levels may lead to phosphate ion accumulation.

[0084] 2.4 Effects of dietary crude protein levels on serum antioxidant markers in Chinese pangolins

[0085] Table 6. Effects of dietary crude protein levels on serum antioxidant capacity of adult Chinese pangolins

[0086]

[0087] Table 6 shows that there were no significant differences in serum total antioxidant capacity (T-AOC), malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD) among the groups (P>0.05). Oxidized glutathione (GSSG) content differed significantly among the groups (P = 0.003), with group A having the highest level, groups C and D significantly lower than group A, and groups B and E falling in between. Catalase (CAT) activity differed extremely significantly among the groups (P < 0.001), with no significant differences among groups A, B, and C, a significant decrease in group D, and the lowest activity in group E, significantly lower than all other groups. Glutathione peroxidase (GSH-Px) activity differed extremely significantly among the groups (P < 0.001), with group A having the lowest activity, group C significantly higher than group A but lower than groups B, D, and E, and no significant differences among groups B, D, and E.

[0088] Changes in the body's antioxidant capacity reflect, to some extent, the animal's health and physiological condition. In this embodiment, group C enhanced the body's antioxidant capacity by reducing GSSG accumulation and increasing GSH-Px activity; while high chitin levels (groups D and E) significantly inhibited CAT activity. Although no significant lipid peroxidation damage was induced (no significant difference in MDA), it may weaken the body's ability to scavenge H2O2. Long-term intake may have potential effects on the body's antioxidant system, indicating that the dietary chitin level setting in this embodiment was reasonable, and group C had stronger antioxidant capacity.

[0089] 2.5 Effects of dietary crude protein levels on apparent nutrient digestibility in adult Chinese pangolins

[0090] Table 7. Effects of dietary crude protein levels on the apparent digestibility of nutrients in Chinese pangolins.

[0091]

[0092] Table 7 shows that the level of chitin in the feed had no significant effect on the apparent digestibility of dry matter (DM), crude protein (CP), and total energy (GE) of Chinese pangolins (P > 0.05). The level of chitin in the feed had a highly significant effect on the apparent digestibility of crude fat (EE), crude ash (Ash), and chitin (P < 0.001). Specifically, the EE digestibility was highest in groups C and D with no significant difference, lowest in group B, and there was no significant difference between groups A and E.

[0093] Both crude ash and chitin digestibility showed a significant decreasing trend with increasing chitin levels in the feed (P < 0.05). Crude ash digestibility was highest in group A, with no significant difference between groups B and C, group D falling between groups C and E, and group E having the lowest digestibility. Chitin digestibility showed no significant difference among groups A, B, and C, but group D was significantly lower than the other three groups and higher than group E, with group E having the lowest digestibility. In summary, appropriate chitin levels can maintain normal digestion and metabolism in adult Chinese pangolins, without interfering with the digestibility and utilization of dry matter, crude protein, and total energy, and can optimize crude fat digestibility. Low chitin levels cannot meet intestinal physiological needs, while high chitin levels lead to an excessive digestive burden and a decrease in both ash and chitin digestibility.

[0094] 2.6 Effects on gut microbiota

[0095] There were no significant differences in gut microbiota richness and diversity indices among the treatment groups (P > 0.05). At the genus level, group C (6%) had the most balanced gut microbiota composition, with the lowest abundance of opportunistic pathogens (Escherichia-Shigella) and beneficial bacteria (Romboutsia, Clostridium sensu stricto 1) maintained at a high level.

[0096] 2.7 Determination of the optimal total chitin level

[0097] The comprehensive evaluation scores of the 15 core difference indicators showed a unimodal distribution: Group C (6%) > Group D (8%) > Group B (4%) > Group E (10%) > Group A (2%). The quadratic polynomial fitting equation is: y = -0.030161χ 2 + 0.389949χ -0.456240, R 2 = 0.9993. The theoretically optimal chitin level is 6.46%. Based on experimental data, 6% is determined to be the optimal total chitin level.

[0098] Conclusion: The suitable total chitin content in the diet of captive adult male Chinese pangolins is 5.5%~6.5%, with 6.00% being preferred.

[0099] Example 2

[0100] Preparation and feeding of the preferred formula

[0101] 1. Formula

[0102] The following raw materials were weighed out by dry matter percentage: bee pupae 9.30%, dried silkworm pupae 6.47%, black ant 30.00%, dried black soldier fly larvae 21.30%, dried mealworms 10.64%, dried crickets 1.30%, dried earthworms 1.70%, housefly larvae 5.30%, sterilized soil 11.92%, and additionally added chitin 2.07%. The sum of all raw materials is 100.00%, and the total chitin content of this formula is determined to be 6.00%.

[0103] 2. Preparation method

[0104] Add the weighed raw materials and water to a blender at a mass ratio of 1:1.8, blend and mix evenly to obtain the final product.

[0105] 3. Feeding method

[0106] Adult male Chinese pangolins were fed once daily at 16:00, allowing them free access to food. The results showed that pangolins fed this formula had significantly better daily feed intake and apparent crude fat digestibility than the 2% and 10% groups, better fecal formation (score 3.13±0.06), optimal serum antioxidant capacity and lysozyme activity, and a healthier gut microbiota.

Claims

1. A diet for adult male Chinese pangolins, comprising the following ingredients in weight percentages: Bee pupae 9.30%, dried silkworm pupae 6.47%, black spiny ant 30.00%, dried black soldier fly larvae 21.30%, dried mealworm 10.64%, dried cricket 1.30%, dried earthworm 1.70%, housefly larvae 5.30%, soil 11.92%, chitin 2.07%.

2. The diet for adult male Chinese pangolins according to claim 1, wherein: The total chitin content of this diet is 5.5% to 6.5%.

3. The diet for adult male Chinese pangolins according to claim 2, wherein: The total chitin content in the diet is 6.0%.

4. A method for feeding adult male Chinese pangolins according to any one of claims 1 to 3, comprising: Adult male Chinese pangolins were fed food and water at a mass ratio of 1:1.8, which were then crushed and mixed evenly. The adult male Chinese pangolins were fed once a day and allowed to eat freely.

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