Tóm tắt Luận án Biological characteristics and effects of gonadotropin on reproductive ability of common palm civets (paradoxurus hermaphroditus pallas, 1777) in captivity

The concentrations of fecal E2 in non-pregnant civets in our

study ranged from 0.05 to 7.01 µg/g df, with an average of 1.07 ± 0.84

µg/g df and a peak of 3.22 ± 0.64 µg/g df. Although faecal estradiol

value of civets have not been published, faecal estradiol values in other

animals have already been widely used to monitor their sexual

activities. For example, faeal E2 of the Siberian tiger population

ranged from 0.39 to 0.49 µg/g and the mean faecal E2 of Bengal tiger

was 0.45 µg/g, and thoat of Sumatra tiger was 2.36 µg/g [89].

Changes in E2 levels showed a cyclic fertilization. The duration

of each cycle ranged from 26.8-33.1 days with the average of 28.6 ±

2.29 days. This period was comparable with that (27.0 days) of

Siberian tigers [89] and of Bengal tigers (29.3 days) [88], but different

from that of domestic cat (21 days) or leopard (10-20 days)[120]

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ical indicators, 14 blood and urine biochemical parameters. All indicators were monitored in normal health status, by age and by gender. 5 2.3.2.3. Blood sampling and analysis - Blood samples were collected through the tail peduncle vein with 3ml syringe (needle size 25Gx1). - Blood physiological indicators were performed by using automated hematology analyzer Mindray BC 2800 Vet. - Blood biochemical parameters were measured by chemical analyzer (Abaxis Vetscan 2, Union City, CA, USA). 2.3.2.4. Urinary sampling and analysis - Urinary samples were collected from the stainless steel floor of a special barn by cylinder, 18-20 hours, 1 time/week for 1 month (for each individual). - Urinary biochemical norms were measured on an automated analyzer (model Teco TC-101, Teco diagnostics, USA). - Na+, K+ and Cl- analyzes were measured from the supernatant obtained by the centrifugation of urine samples from the civets at 3000 rpm for 10 min (Roto x 32®-Hettich) and were performed on an ion selective device (model Roche 9180, Roche Diagnostics, Switzerland). 2.3.3. Study on changes in sex hormone content of female civets in captivity 2.3.3.1. Materials Fecal samples were collected from 12 mature female civets for 16 months. 2.3.3.2. Survey targets Changes in the endocrine index: estradiol (E2), progesterone (P4) of adult female civets in the following cases: non-pregnant, pregnant and pseudopregnancy. 2.3.3.3. Fecal sampling and extraction 6 Fecal samples were collected an average of 3 days, about 18h - 20h in 14 months. Fresh samples (5g) were collected in a plastic bag and stored (-200C) until analysis. After thawing, 0.2 g was weighed and placed in a glass jar containing 2 ml of 90% methanol. After shaking for 30 min, the suspension was centrifuged at 1,700 x g for 20 min (on a shaker EAB 20). After centrifugation, approximately 1 ml of the aqueous solution was extracted into 1.5 ml eppendorf vial and frozen (-200C) until using (Frederick et al., 2010). The remainder is returned to the glass vial and dried to determine the dry weight of the stool. 2.3.3.4. Hormone assays The fecal contents of P4 and E2 were determinned by using fully automatic ELISA Dynex DS2 (Dynex, USA), Progesterone and Estradiol ELISA Kit. 2.3.4. Study the effect of gonadotropin (PMSG, HCG) on reproductive ability of the civets 2.3.4.1. Materials A total of 54 mature female civets, after the survey were classified into 3 groups: - Group 1: Civets were slow to breed for the first time (after 24 months of age, there was no estrus manifestation); n = 14. - Group 2: Slowly rejuvenating civets (after 12 months after birth, no estrus have been seen); n = 15. - Group 3: Low productivity breeding civets (1 litter/year; number of civets per litter was small, 1-2 newborn/litter); n = 25. 2.3.4.2. Survey targets - Changes in endocrine indexes: estradiol (E2), progesterone (P4) after injection of PMSG and HCG (from 2 days before injection (day 7 -2) to the injection date (day 0) and 8 days later treatment. - The duration of the civets’ oestrus after injection of PMSG and HCG. - Ratio of estrus civets (estrus civets/total number of treated civets). - Percentage of pregnancy civets (number of pregnant civets/total number of treated civets). - The average of newborn civets on the litter. - Average weight of newborn civets. - The proportion of live civets after 24 hours and after 1 month (number of surviving civets /total number of newborn civets). 2.3.4.3 Type of reproductive hormones Mixed PMSG / HCG (ratio 2: 1): trade name is Gestavet (United Kingdom). Each vial contained 400 IU PMSG/200 IU HCG and vial containing 5ml solvent for injection solution. 2.3.4.4. The treatments for injecting reproductive hormones Control (ĐC): 0 IU PMSG + 0 IU HCG (No injection) Treatment 1 (CT1): 20 IU PMSG + 10 IU HCG Treatment 2 (CT2): 30 IU PMSG + 15 IU HCG Treatment 3 (CT3): 40 IU PMSG + 20 IU HCG The experiment was performed all treatments for civets in 3 groups. Each batch has 3 or more civets. 2.3.4.5. Experimental layouts Table 2. 2. Experimental layout diagram Group ĐC CT1 CT2 CT3 Group 1 (n=14) 3 4 4 3 Group 2 (n=15) 3 4 4 4 Group 3 (n=25) 3 7 7 8 2.3.4.6. Injection process 8 The female civets are injected intramuscularly (IM) at 8 AM. 2.3.4.7. Methods of determining hormone changes Sampling procedure, stool extract, hormone test is done similarly 2.3.3.3 and 2.3.3.4. 2.3.4.8. Method of determining oestrus - Observation method. - Clinical method: monitoring pregnancy and childbirth of female civets. 2.4. Data analysis From the data obtained, conduct the calculation of statistical parameters: average value (X̅), standard deviation (SD); ANOVA analysis of a factor with a level of α = 0.05. Calculations were processed by MS-Excel 2013 software. CHAPTER 3. RESULTS AND DISCUSSION 3.1. Results of studying biological characteristics 3.1.1. Morphological characteristics and some behaviors of civets in culture conditions In terms of morphology, the civets in the captivity also has the characteristics of the species as in natural conditions. The coat is gray or moldy, sucking black fur; Vertebral vertebrae, black-brown ribs or often forming three stripes along the spine from the shoulders to the base of the ears. The tail has unknown streaks or is black at the base of the tail, the tail is usually black, but in some civets it may be white; the nose, cheeks, ears, lower thighs and four legs are black; gray belly. When the civets are still young, the black brown stripe is not clear, the hair is rough. The more you grow, the smoother the fur and the 9 dark brown the clearer. The face has 2-3 bright spots on the forehead or the edge of the eyes. Regarding nutritional characteristics, the civets are omnivorous. Survey results showed that civets eaten many fruits. The favorite fruit of civets is bananas. In breeding, depending on the season, it is possible to change different food sources. In terms of animal feed, civets especially like to eat meat, fish and eggs. According to Dang Huy Huynh et al., (2010); Duckworth (2016), the civets is omnivorous, they can eat most of the food that humans eat [7, 111]. The civets mainly act at night, sleep at day. Excretion activities are mainly carried out at the beginning of the evening operation phase. Civets has very high protective behavior. When grazing another child into a barn, they will bite each other. The civet is only paired when the offspring has arousal. 3.1.2. Growth characteristics of the civet civet in captivity 3.1.2.1. Volume growth Survey results of mass growth rate of 64 civets were shown in Table 3.1. Table 3. 1. Growth rate of the civets’ weight in captivity Table 3.1 showed that the amount of civets that was monitored has an uneven growth rate over the ages of months, which was consistent with the periodic growth rules. Volume growth tended to increase gradually from 3-12 months of age, then gradually slow down X̅(g) Sx Cv% A (g/head/day) Rw% X̅(g) Sx Cv% A (g/head/day Rw% 3 782 118.5 15.15 727 82.7 11.38 55 >0,05 6 1,152 126.1 10.95 4,11 a 38,26 a 975 125.3 12.85 2,76 b 29,14 b 177 <0,05 9 1,735 109.1 6.29 6.48 40.39 1,456 117.8 8.09 5.34 39.57 279 <0,05 12 2,644 113.5 4.29 10.10 41.52 2,225 113.7 5.11 8.54 41.78 419 <0,05 15 3,245 128.1 3.95 6.68 20.41 2,848 120.7 4.24 6.92 24.56 397 <0,05 18 3,533 108.5 3.07 3.20 8.50 3,175 118.9 3.74 3.63 10.86 358 <0,05 21 3,743 116.3 3.11 2.33 5.77 3,335 84.7 2.54 1.78 4.92 408 <0,05 24 3,925 105.2 2.68 2.02 4.75 3,516 93 2.63 2.01 5.28 409 <0,05 X̅ 5.14 20.22 4.71 21.16 Age (month) Male (n=32) (1) Female (n=32) (2) X̅1 - X̅2 P 10 from 12-24 months. The period with the highest growth rate was from 9 to 12 months with the average absolute growth of each male being 10.1 g/head/day, growing relatively R% = 41.52%, These indicators corresponded to females of 8.54 g/head/day and R% of 41.78%. 3.1.2.2. Growth in body length The survey results of growth of civets’ body length were shown in Table 3.2. Table 3 2. Growth rate of body length of the civets in captivity Table 3.2 showed that the growth rate of the civets’ body length was slow down through the months of age. The fastest growth phase of body length was the period of 3-6 months with absolute growth of 2.87 cm/head/month, relative growth R% = 19.46% (for the period of 3 months) in males and 2.57 cm/head/month, relative growth of R% = 18.15% in females. After the 15th month, when the stem length was close to the good limit of the species, the growth in the period of 18-24 months was very small (0.10-0.76 cm/head/month). 3.1.2.3. Growth in tail length The survey results of the civets’ tail length growth were shown in Table 3.3. Table 3. 3. Growth rate of the civets’ tail length in captivity X̅(g) Sx Cv% A (cm/head/m) Rw% X̅(g) Sx Cv% A (cm/head/m) Rw% 3 39.89 0.38 0.95 38.67 0.32 0.83 1.22 >0,05 6 48.49 0.44 0.91 2.87 19.46 46.39 0.46 0.99 2.57 18.15 2.10 >0,05 9 56.27 0.46 0.82 2.59 14.85 53.24 0.53 1.00 2.28 13.75 3.03 <0,05 12 62.55 0.55 0.88 2.09 10.57 59.01 0.55 0.93 1.92 10.28 3.54 <0,05 15 68.92 0.45 0.65 2.12 9.69 65.12 0.54 0.83 2.04 9.84 3.80 <0,05 18 70.36 0.77 1.09 0.48 2.07 67.41 0.31 0.46 0.76 3.46 2.95 <0,05 21 71.33 0.41 0.57 0.32 1.37 68.68 0.29 0.42 0.42 1.87 2.65 <0,05 24 71.62 0.41 0.57 0.10 0.41 69.03 0.30 0.43 0.12 0.51 2.59 <0,05 X ̅ 1.51 8.35 1.45 8.27 Age (month) Male (n=32) (1) Female (n=32) (2) X̅1 - X̅2 P X̅(g) Sx Cv% A (cm/head/m) Rw% X̅(g) Sx Cv% A (cm/head/m) Rw% 3 36.44 0.93 2.55 36.02 1.18 3.28 0.42 >0,05 6 38.24 0.95 2.48 0.60 4.82 37.68 1.28 3.40 0.55 4.50 0.56 >0,05 9 41.45 0.91 2.20 1.07 8.06 40.82 1.18 2.89 1.05 8.00 0.63 >0,05 12 45.36 0.91 2.01 1.30 9.01 44.68 1.33 2.98 1.29 9.03 0.68 >0,05 15 48.64 0.91 1.87 1.09 6.98 48.19 1.34 2.78 1.17 7.56 0.45 >0,05 18 51.37 0.87 1.69 0.91 5.46 50.88 1.22 2.40 0.90 5.43 0.49 >0,05 21 54.25 0.80 1.47 0.96 5.45 53.57 1.25 2.33 0.90 5.15 0.68 >0,05 24 56.31 0.78 1.39 0.69 3.73 55.62 1.03 1.85 0.68 3.75 0.69 >0,05 X ̅ 0.95 6.21 0.93 6.20 Age (month) Male (n=32) (1) Female (n=32) (2) X̅1 - X̅2 P 11 Table 3.3 shows that the growth of the tail length of the civets increased relatively evenly over the period of age, however, increased faster in the period of 6-12 months (R% from 8-10, 14%). The absolute growth rate (A) averageed 0.95 cm/head/month, the relative growth rate (R%) was 6.21% (in males) and A = 0.93 cm/head/month, R% = 6.20% (in females). 3.1.2.4. Chest growth The results of monitoring the dimension of chest girth of the civets in captivity were shown in Table 3.4. Table 3. 4. Growth rate of the civets’ chest girth in captivity Table 3.4 showed that the chest girth growth rate was high in the period of 3-12 months of age and highest in the 9-month period (A = 1.27 cm/head/month, R = 15.07% in males) and A = 1.06 cm/ head/month, R = 13.21% in females). This was in accordance with the rules of fast chest growth rate in the sexual maturity stage in animals. Age of sexual maturity of civets was in the period of 9-12 months of age [111]. 3.1.3. Reproductive characteristics of incense civet in captivity 3.1.3.1. Age of sexual maturity and estrus manifestations Results of monitoring the age of sexual maturity of civets (32 female civets, 34 male civets) in captivity in the study were shown in Table 3.5. X̅(g) Sx Cv% A (cm/head/m) Rw% X̅(g) Sx Cv% A (cm/head/m) Rw% 3 21.21 0.92 4.34 20.83 0.81 3.89 0.38 >0,05 6 23.43 0.83 3.54 0.74 9.95 22.56 0.71 3.15 0.58 7.97 0.87 >0,05 9 27.25 0.75 2.75 1.27 15.07 25.75 0.70 2.72 1.06 13.21 1.50 <0,05 12 29.36 0.93 3.17 0.70 7.45 27.64 0.70 2.53 0.63 7.08 1.72 <0,05 15 29.87 0.92 3.08 0.17 1.72 28.24 0.74 2.62 0.20 2.15 1.63 <0,05 18 30.32 0.90 2.97 0.15 1.50 28.68 0.66 2.30 0.15 1.55 1.64 <0,05 21 30.54 0.86 2.82 0.07 0.72 28.93 0.82 2.83 0.08 0.87 1.61 <0,05 24 30.66 0.85 2.77 0.04 0.39 29.12 0.78 2.68 0.06 0.65 1.54 <0,05 X ̅ 0.45 5.26 0.39 4.78 Age (month) Male (n=32) (1) Female (n=32) (2) X̅1 - X̅2 P 12 Table 3. 5. Age of sexual maturity of the civets in captivity Note: Different characters in the same row are statistically significant differences (P <0.05). The table 3.5 showed that the female civets were starting to show oestrus in the period of 10-14 months with an average weight of 2.38 - 2.62 kg. The average maturity was 11.96 months, with an average weight of 2.50 kg. According to Nelson (2013), the estrus of civets was between 11 and 12 months of age [112]. 3.1.2.2. Mating activity, pregnancy rate and pregnancy time During the study period, we monitored the reproductive results of 42 female civets, with 84 pairings. Results of monitoring pregnancy rate and gestation period were presented in Table 3.6. Table 3. 6. Pregnancy ratio and duration in the civets Note: the characters in the same column are different, the difference is statistically significant (P <0.05). Age (month) Number Rate (%) Weight (kg) Age (month) Number Rate (%) Weight (kg) 9 0 0 9 2 5.88 1.84 10 4 12.50 2.38 10 9 26.47 2.45 11 5 15.63 2.43 11 14 41.18 2.56 12 9 28.13 2.51 12 6 17.65 2.61 13 6 18.75 2.58 13 3 8.82 2.86 14 3 9.38 2.62 14 0 0.00 Not mature 24-30 5 15.63 3.26 0 0 X̅ 11,96a 2,50 10,97b 2,52 SD 1.22 0.08 1.03 0.04 Parameter Female (n=32) Male (n=34) Mature Place No of mating civets (n=84) No of pregnancy civets (heads) R (%) Gestation length time (days) Dong nai 30 14 46,67a 61,2 Thu Duc 54 42 77,78b 60,8 X̅ 66.67 60.9 SD 1.3 13 3.1.3.3. Number of births per litter, survival rate, neonatal characteristics and weaning age Results of monitoring the number of young civets on litter, neonatal characteristics and survival rates of 56 monitored parities were presented in Table 3.7. Table 3 .7. The number of piglets born on the litter and the survival rate Note: the characters in the same column are different, the differences are statistically significant (P <0.05). Table 3.7 showed that the births of each litter were range from 1 - 4. The average number of civets per litter of all 56 litters was 2.38. In nature, according to Dang Huy Huynh et al., (2010), civets lay 2 to 4 newborns [7]. According to Nelson (2013), 2-5 newborns, an average of 3.4 newborn civets /litter [112]. Newborn civets were very small and weak, unable to stand firmly, with an average weight of 95.16 grams. After a period of 7-10 days, the rim of the ear opened, from 12-15 days the eyes opened. At birth 24h 48h 1 week 1 month At Weaning Weight at birth Weight at weaning 1 4 2 6 3 3 4 1 X̅ 1 1,86a 96,93 585,36 SD 0.77 7.45 26.85 1 6 2 15 3 14 4 7 X̅ 2 2,55b 94,57 59142 SD 0.92 6.27 19.38 X ̅ 2.38 96.15 90.03 87.22 81.53 80.83 80.83 95.16 590.83 Dong Nai (n= 14) 22 20 20 17 Place Number of newborn civets (con) Number of (con) n=56 Live newborn civets (n / %) Weight (X̅ , gr) 17 17 96.93 585.36 84.62 76.92 76.92 65.38 65.38 65.38 Thu Duc (n=42) 107 101 97 93 92 94.57 592.65 100.00 94.39 90.65 86.92 85.98 85.98 92 14 3.2. Research results of some physiological and biochemical indicators of blood of civets in captivity 3.2.1. Hematological parameters of common palm civets by gender The results of the study on the physiological parameters of 186 blood samples from 62 civets (30 males and 32 females) were presented in Table 3.8. Table 3.8.Hematological data of common palm civets by gender group 15 3.2.2. Hematological parameters of common palm civets by age The results of the study on the physiological parameters of 186 blood samples from 62 civets by age were presented in Table 3.9. Table 3.9. Hematological data of common palm civets by age group 3.2.3. Serum biochemical parameters of common palm civets The results of blood biochemical parameters by sex and gender group were shown in Table 3.10. and Table 3.11. Table 3.10. showed that the total serum protein (TP), globulin, albumin and phosphorus levels in males were higher than females, the difference was statistically significant (P <0.05). 16 Table 3.10. Serum biochemical parameter of commom palm civets by sex group Table 3. 11. Serum biochemical parameter of commom palm civets by age group 17 3.2.4. Urinary biochemical parameters of common palm civets by sex The results of the study on the urinary biochemical parameters from 60 civets were presented in Table 3.12. Table 3.12. Urinary biochemical data of commom palm civets by sex group 18 3.2.5. Urinary biochemical parameters of common palm civets by age The results of urine biochemical indexes of civets in the age group were presented in Table 3.13. Table 3.13. Urinary biochemical data of commom palm civets by age group Parameter 3-12 months (n=31) Mean ± SD Mean ± SD Weight (g) 1.735±109,1a 3.335 ±84,7b Body legth (mm) 56,27±0,46a 71,62±0,41b Urobilinogen (µmol/L) 10,44 ±1,06 10,88 ±1,27 Glucose (mmol/L) Neg1a 0,47±0,22b Billirubin (µmol/L) 0,46±0,06 0,35±0,05 Ketone (mmol/L) 0,13±0,02 0,17±0,03 Specific Gravity 1,02±0,01 1,02±0,01 Blood (Ery/µL) neg neg pH 7,55±0,17 7,51±0,52 Protein (g/L) 16,01±1,27 15,88±1,31 Nitrite neg neg Leukocytes (Leu/µL) 3,82 ± 0,12a 5,35 ± 0,37b Ascorbic acid (mmol/L) 0,15±0,03 0,17±0,05 K (mmol/L) 173,23±43,12 185,27±51,25 Na (mmol/L) 69,86±11,07a 83,75±16,32b Cl (mmol/L) 149,58±43,42 157,47±32,24 19 3.4. The results on the changes of sex hormone of female civets in captivity 3.4.1. The changes of estradiol and progesterone in non-pregnant civets 3.4.1.1. Faecal estradiol in non-pregnant civets The concentrations of fecal E2 in non-pregnant civets in our study ranged from 0.05 to 7.01 µg/g df, with an average of 1.07 ± 0.84 µg/g df and a peak of 3.22 ± 0.64 µg/g df. Although faecal estradiol value of civets have not been published, faecal estradiol values in other animals have already been widely used to monitor their sexual activities. For example, faeal E2 of the Siberian tiger population ranged from 0.39 to 0.49 µg/g and the mean faecal E2 of Bengal tiger was 0.45 µg/g, and thoat of Sumatra tiger was 2.36 µg/g [89]. Changes in E2 levels showed a cyclic fertilization. The duration of each cycle ranged from 26.8-33.1 days with the average of 28.6 ± 2.29 days. This period was comparable with that (27.0 days) of Siberian tigers [89] and of Bengal tigers (29.3 days) [88], but different from that of domestic cat (21 days) or leopard (10-20 days)[120]. 3.4.1.2. Faecal progesterone in non-pregnant civets In this study group, non-pregnant female civets’ fecal progesterone (P4) metabolites levels ranged from 0.15 to 12.32 µg/g df with the overall mean of 1.72 ± 2.16 µg/g df . For comparison, faecal progesterone level of Siberian tigers varies from 0.27 to 38.19 µg/g and that of the Sumatra tigers ranged from 0.09 to 18.52 µg/g, and the level in Bengal Tigers was 36.05µg/g [89]. Faecal progesterone levels of civets also changed over time. The peak of faecal progesterone level ranged from 6.03-12.32 µg/g df with an average of 7.26 ± 1.11 µg/g df. The cycle of change in progesterone level ranged from 26.6 to 31.0 days with an average of 27.8 ± 2.80 days. 20 3.4.2. The changes of estradiol and progesterone in pregnant civets During pregnancy, the civets’ faecal P4 level ranged from 6.21 to 23.12 µg/g df with an average of 15.17 ± 5.22 µg/g df. This value was approximately 5 to 7 times higher (P <0.05) than non-pregnant and post-fertilization periods. In individuals with conception, P4 increased significantly for a period between 60 and 63 days after fertilization. Faecal E2 concentration of a juvenile fennel during pregnancy was relatively lower than that in other periods. Fecal E2 ranged from 0.22 to 1.05 µg/g df with an average of 0.74 ± 0.23 µg/g df. After parturition, E2 increased and marked the recovery of ovarian activity from 25-30 days. In contrast, in fetal predisposed individuals, there was a negligible E2 change (range of 0.35-1.99 µg/g df) compared to non-pregnant ones (P>0.05), and distinctly lower than that of the pregnancy period (P <0.05). This result was similar to that observed in the Pallas' cat, the clouded leopard, Tiger, in these an increase in estrogen excretion in faeces was not observed during pregnancy [120]. 3.4.3. The changes of estradiol and progesterone in pseudopregnancy civets In pseudopregnancy civets, there were also significant changes in P4 after fertilization, but the change duration was only 26- 30 days. The faecal P4 level at this stage ranged from 8.02 to 11.47 µg/g df with an average of 9.73 ± 1.73 µg/g df. This value was significantly higher than that of non-pregnant civets but was significantly lower than those of pregnant animals (P<0.05). In other studies, leopard cats, clouded leopards, snow leopards and cheetahs have been reported to have increased duration of P4 contents during presumed pseudopregnancy [89]. Thus, the main indicator to distinguish between pregnancy and fake pregnancies is both the duration and the degree of the increase in the fecal P4. 21 3.5. Results of studying the effects of sex hormones (PMSG, HCG) on fertility of the civets 3.5.1. The changes of estradiol and progesterone after injecting sex hormones Results of monitoring E2 and P4 changes showed that E2 content in the feces of the civets in the treatments began to increase at day 1 after injection, peaking on day 2 and then gradually decreasing from day to day. 3rd, the average E2 content of day 2 when peaking in formula 1 was 2.59 µg/g df, treatment 2 was 2.69 µg / g df and treatment 3 was 3.16 µg/g df ; in which the highest was in treatment 3. Thus, when the higher treatment dose (40IU PMSG) increased the amount of sex hormones in the treated civet group (P <0.05). Meanwhile in the control group, E2 content in the stool changed over the days, but this change was not significant (P> 0.05). By the 8th day, the hormone content in the stool in all treatments decreased to close to the value before sexual stimulation (day -2 and day -1). 3.5.2. Time appears manifestations and prolonged estrus In all experimental groups, the average duration of oestrus was 1.1 days (CT3 in group 2) to 2.6 days (CT1 in group 2 and group 3) after injection of sex hormones. In particular, CT3 always has the earliest time of estrus (from 1-1.5 days). The duration of oestrus was strongly correlated with the peak time of E2 (R = 0.82). The duration of oestrus in all formulas ranged from 2.9 to 3.9 days, of which the shortest time was in CT1 and the longest was in CT3. This time was longer than the time of expression of oestrus in the civet without sexual stimulation during the breeding season (2-3 days). ANOVA analysis showed that there was a statistical difference (P <0.05) on the duration of oestrus and duration of oestrus between the formulas in each experimental civet group. CT3 always has the earliest occurrence of oestrus and the longest period of oestrus. 22 3.5.3. The results of using hormone formulas on some reproductive indicators Results of monitoring reproductive efficiency after injection of sex hormones were presented in table 3.19. Table 3. 19. Results of reproductive performance monitoring after injecting sex hormones Treatment ĐC CT1 CT 2 CT 3 n 9 15 14 15 Estrus civets Civets 1 12 12 14 % 11,11a 80,00b 85,71c 93,33d Pregnant civets Civets 1 10 10 12 % 11,11a 66,67b 71,43c 86,67d Newborn/ litter (X̅ ± SD) Civets/litter 2±0a 3,04± 0,47b 3,41± 0,66c 3,53± 0,90c Weight at birth (X̅ ± SD) (g) 95,5±0 96,15± 2,14 95,72± 2,17 94,02± 3,35 Survivall rate after 48h (%) 100,00a 86,60b 86,35b 90,00b Survivall rate after 1 month (%) 100,00a 83,57b 82,57b 82,83b Note: The difference of the characters in the same row is statistically different (P <0.05), according to the T-test test with the significance level α = 0, 05. CONCLUTIONS AND RECOMMENDATIONS 1. Conclusion 1.1. Biological characteristics of the civets in captivity - Description of morphological characteristics, nutritional characteristics, weight growth, body length, tail length and chest girth of the civets in the period of 3-24 months. - Identified so

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