| 区炳明，王嘉仪，李清青，林 雪，汤晓惠，钟炜楠，周月育，李菊琴.仔猪阶段使用过抗生素的陆川猪母猪肠道和乳汁中大肠杆菌的耐药性及毒力基因分析[J].广东农业科学,2024,51(9):30-40 |
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仔猪阶段使用过抗生素的陆川猪母猪肠道和乳汁中大肠杆菌的耐药性及毒力基因分析 |
| Analysis of Antibiotic Resistance and Virulence Genes in Escherichia coli Isolated from the Intestine and Milk of Luchuan Sows Treated with Antibiotics during the Piglet Stage |
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| DOI：10.16768/j.issn.1004-874X.2024.09.003 |
| 中文关键词: 母猪 肠道 乳汁 大肠杆菌 耐药性 毒力基因 |
| 英文关键词: sow intestinal tract milk Escherichia coli antibiotic resistance virulence gene |
| 基金项目:国家自然科学基金（32102703）；肇庆学院农村科技特派员工作站项目（2022N001）；肇庆学院大学
生创新创业培训计划项目（X202310580123）；广东省大学生创新创业培训计划项目（S202410580044） |
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| 中文摘要: |
| 【目的】尽管替抗减抗政策已出台多年，但为预防和治疗疾病，在仔猪早期使用抗生素仍然是集约
化养殖场养殖管理的一个重要环节。研究养殖场中仔猪阶段使用过抗生素的母猪体内不同部位（肠道和乳腺）大肠
杆菌的耐药性和毒力差异情况，为集约化养殖场抗生素使用的优化提供科学依据。【方法】记录广东省湛江市某集
约化陆川猪场母猪从仔猪出生后至约 420 日龄的抗生素使用情况，从同一批次约 420 日龄的健康母猪中收集初乳（分
娩后 2 h 内）、常乳（分娩后 24~72 h 内）及肛门内新鲜粪便样品，通过麦康凯鉴别培养基分离出疑似大肠杆菌。
通过药敏试验分析大肠杆菌耐药性和 PCR 检验毒力基因存在情况。【结果】通过麦康凯鉴别培养基分离出疑似大
肠杆菌共 704 株，其中粪便大肠杆菌 300 株、初乳大肠杆菌 220 株、常乳大肠杆菌 184 株。药敏试验显示，粪便大
肠杆菌对氨苄西林和四环素的耐药率为 96.3%（289/300），对其他常规抗生素的耐药率为 63.6%（7/11），总耐药
率 98.7%（296/300）；初乳大肠杆菌对氨苄西林和四环素的耐药率为 99.5%（219/220），对其他常规抗生素的耐
药率为 100%（1/1），总耐药率 100%（220/220）；常乳大肠杆菌对氨苄西林和四环素的耐药率为 40.8%（75/184），
对其他常规抗生素耐药率为 67.9%（74/109），总耐药率 81.0%（149/184）。最终鉴定出 39 株无耐药性的大肠
杆菌。PCR 检测显示， 39 株无耐药性的大肠杆菌中大多数不含猪致病性大肠杆菌常见毒力基因（占比 89.74%，
35/39）。此外，无耐药性常乳大肠杆菌中含毒力基因的菌株占比仅为 2.86%（1/35），无耐药性粪便大肠杆菌中含
毒力基因的菌株高达 75.0%（3/4）。【结论】仔猪阶段使用过抗生素防控病害的陆川猪，到生长成初产母猪后，
从其肛门和乳腺中分离的大肠杆菌菌株中仍存在少数无耐药性且无毒力基因的大肠杆菌菌株。其中，粪便中的无耐
药性且无毒力基因大肠杆菌占比极少，仅为 0.33%（1/300）；分娩 2 h 内的初乳中未能分离出无耐药性且无毒力基
因的大肠杆菌，占比 0%（0/220）；但分娩后 24~72 h 内的常乳中仍存在较高比例的无耐药性且无毒力基因的大肠
杆菌，占比 18.48%（34/184）。 |
| 英文摘要: |
| Abstract: 【Objective】Although the policies for reducing or replacing antibiotics have been in place for many years,
the use of antibiotics in the early stages of piglet rearing remains an important aspect of disease prevention and treatment in
intensive farming operations. The study of the differences in antibiotic resistance and virulence of Escherichia coli isolated
from different parts (intestinal tract and mammary gland) of sows with antibiotics in piglet stage in farms can provide a scientific
basis for the optimization of antibiotic use in intensive farms.【Method】This study recorded the antibiotic usage of sows in
a intensive Luochuan pig farm in Zhanjiang City, Guangdong Province, from the birth to around 420 days of age of piglets.
Colostrum (within 2 h after delivery), regular milk (24-72 h after delivery) and fresh fecal samples from the anus were collected
from healthy sows of the same batch at approximately 420 days of age. Suspected E. coli isolates were obtained by using
MacConkey agar plates. Antibiotic susceptibility was analyzed through antibiotic sensitivity test, and the presence of virulence
genes was examined by PCR.【Result】A total of 704 suspected E. coli isolates were obtained by using MacConkey agar plates,
including 300 isolates from fecal samples, 220 from colostrum, and 184 from regular milk. Antibiotic sensitivity test revealed
that the resistance rate of fecal E. coli to ampicillin and tetracycline was 96.3% (289/300), with a resistance rate of 63.6%
(7/11) to other common antibiotics, and an overall resistance rate of 98.7% (296/300). The resistance rate of colostrum E. coli
to ampicillin and tetracycline was 99.5% (219/220), with a resistance rate of 100% (1/1) to other common antibiotics, and an
overall resistance rate of 100% (220/220). The resistance rate of regular milk E. coli to ampicillin and tetracycline was 40.8%
(75/184), with a resistance rate of 67.9% (74/109) to other common antibiotics, and an overall resistance rate of 81.0% (149/184).
A total of 39 non-antibiotic-resistant E. coli strains were identified. PCR detection showed that most of the 39 non-antibioticresistant E. coli strains did not contain common virulence genes associated with porcine pathogenic E. coli (89.74%, 35/39).
Additionally, only 2.86% (1/35) of non-antibiotic-resistant regular milk E. coli strains carried virulence genes, while 75.0% (3/4)
of non-antibiotic-resistant fecal E. coli strains carried virulence genes.【Conclusion】From the stage where antibiotics are
used to prevent and control diseases in piglets to their growth into primiparous sows, a small number of non-virulent and nonantibiotic-resistant E. coli strains are still isolated from the anus and mammary glands. Among them, the proportion of nonvirulent and non-antibiotic-resistant E. coli in feces is extremely low, accounting for only 0.33% (1/300). No non-virulent and
non-antibiotic-resistant E. coli are isolated from colostrum within 2 hours after delivery, accounting for 0% (0/220). However, a
relatively higher proportion of non-virulent and non-antibiotic-resistant E. coli is found in regular milk collected 24-72 h after
delivery, accounting for 18.48% (34/184). |
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