All of the procedures conducted in this study were in accordance with the Guide for the Animal Care and approved by the Institutional Animal Care and the Use Committee of the Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, China.

The experiment was carried out from March to July 2018 in a small-scale local farm based on 200 does (the Xiangdong black goat, a local meat breed) in Liuyang, Hunan Province, China. The average ambient temperature during the study was 25°C (range: 18–31°C). A group housing system consist of half-open sheds featuring with no walls on one side and the size was 20 ± 4 animals per shed. Levamisole (8 mg/kg BW) was monthly mixed into the feed for expelling parasite in the local farm. Mature goats except for pregnant ewes received additional ivermectin at its recommended dose of 0.2 mg/kg BW by subcutaneous injection to further expelling parasite in annual March and September. Goats are vaccinated (Combine Ovine/Caprine Braxy, Struck, Lamb Dysentery and Enterotoxaemia Vaccine, inactivated; Chongqing Auleon Biologicals Co., Ltd., China) in annual mid-February, except for goat kids under 2 months of age and does in late pregnancy.

In later pregnancy, does were fed twice per day at 08:30 and 17:30 with concentrate (per kg DM basis: 74.1 g whey powder, 211 g corn flour, 320 g soybean meal, 65 g fish meal, 220 g fat powder, 51 g milk powder, 8.6 g CaCO3, 25.3 g CaHPO4, 5 g NaCl, and 20 g premix) to meet the normal physiological requirements and grazed for 6 h daily. The premix (per kilogram) was composed of 119.0 g MgSO4·H2O, 2.5 g FeSO4·7H2O, 0.8 g CuSO4·5H2O, 3.0 g MnSO4·H2O, 5.0 g ZnSO4·H2O, 10.0 mg Na2SeO3, 40.0 mg KI, 30.0 mg CoCl2·6H2O, 95,000 IU vitamin A, 17,500 IU vitamin D, and 18,000 IU vitamin E. The forage was fed ad libitum with the same hay composed of twitch grass (dominant), ryegrass, and clover. One week before expected delivery, all pregnant dams were moved from the group housing system and housed in conventional individual pens with straw bedding located within an indoor animal facility with an average temperature of 24 ± 1°C and natural lighting. Straw beddings were replaced once a month.

The kids were ear tagged and individually reared together with their mothers in pens. The feeding trough was accessible at all times and the kids could also eat from the trough. All does and kids had free access to clean water throughout the entire experimental period. Three different investigators with extensive work experience in farm were jointly involved to judge the health of the kid goats. Health scores were evaluated by using fecal scores, nasal scores and attitude scores adapted from the University of WisThe model used for the analysis was as followsconsin calf health scoring chart (https://fyi.extension.wisc.edu/heifermgmt/files/2015/02/calf_health_scoring_chart.pdf). Nasal scores were categorized as 0: no discharge; 1: a small amount of cloudy discharge from one nostril; 2: cloudy discharge from both nostrils; and 3: excessive thick cloudy discharge from both nostrils. Fecal scores were categorized as 0: normal; 1: semi-formed, pasty; 2: loose, but stays on top of bedding; and 3: watery, sifts through bedding. Attitude scores were 0: active; 1: dull; 2: depressed; and 3: no response. Both a nasal score ≥ 1 and a fecal score ≥ 1 considered as diarrhea. And each individual aspect received a score 0 was considered as healthy.

Numerous studies have reported the highest incidence of diarrhea is the first 15 days of life (16, 17). Twin kid goats from the same does (one healthy and the other diagnosed as diarrhea) with the same age under 14 days after birth were used in this study, and a total of 10 pairs were successfully matched. The basic information of experimental kids was shown in Table 1. Once the goat kids were successful matched based on clinical experience, animals were allocated to two groups (diarrhea and healthy control experiment; n = 10/group) and euthanized at the same age and in a same way by intravenous overdose of sodium pentobarbital (50 mg/kg BW) and then started the slaughter test.

The basic information of experimental goat kids.

C means control group; D means diarrhea group; BW means birth weight; SBW means slaughter body weight; NA means data is missing.

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