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標題: 台灣動物臨床分離株對黏菌素與碳青黴烯類抗生素抗藥性之流行病學研究
Epidemiology of Resistance to Colistin or Carbapenems in Clinical Isolates from Animals in Taiwan
作者: 劉芊懿
Qian-Yi Liu
關鍵字: mcr-1;mcr-3;碳青黴烯類抗生素抗藥性;mcr-1;mcr-3;Carbapenem resistance
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多重抗藥性細菌已在全球皆有發現會引起嚴重院內感染,在治療與控制上皆不易,且死亡率高。黏菌素是對抗革蘭氏陰性菌的最後一線藥物之一,特別是針對碳青黴烯類抗生素抗藥性的腸桿菌科所引起人類院內感染症的治療。但在2015年,中國首次發現質體介導黏菌素抗藥基因 (mcr-1),隨後在超過30個國家不論是野生動物、人類或食物上皆有陸續發現,其中包括台灣。
台灣地區對於動物來源的質體介導黏菌素抗藥基因相關文獻十分稀少,而本研究主要目的是調查台灣產食經濟動物、野生動物及伴侶動物所分離之臨床分離株mcr-1的盛行率及抗藥性概況。本試驗樣品來自國立中興大學獸醫學院動物疾病診斷中心2016年1月至2017年6月及國立中興大學獸醫教學醫院2016年1月至2017年4月的臨床分離株檢測其黏菌素抗藥基因,共蒐集了247個不重複的革蘭氏陰性菌臨床分離株,檢測全部分離株是否帶有質體介導黏菌素抗藥基因,其中包括了mcr-1、mcr-2、mcr-3、mcr-4及mcr-5。結果顯示從豬隻、家禽、野生動物及伴侶動物對於黏菌素抗藥的比例分別為33% (42/126)、43% (20/47)、24.3% (9/34)及8% (3/37),而mcr-1之陽性率分別為27.0% (34/126)、21.3% (10/47)、8% (3/37)及2.7% (1/37)。另外,在3株來自豬隻分離株以及1株來自家禽分離株檢測出mcr-3陽性,而同時帶有mcr-1及mcr-3的分離株有3株在豬隻及家禽皆有分離到,而mcr-2、mcr-4及mcr-5並沒有在任何分離株所檢測到。
對48株mcr-1陽性的革蘭氏陰性菌,其中包括了45株大腸桿菌與Aeromonas trota、Acinetobacter baumannii及Salmonella Enteritidis各一株,去除了A. trota,深入探討發現全部mcr-1陽性分離株對於黏菌素、ampicillin及tetracycline都具有抗藥性,90%以上對於trimethoprim-sulfamethoxazole及第一代頭孢子素抗生素 (cefazolin) 具有抗藥性,及35.4%之mcr-1陽性菌株會產生廣效性乙內醯胺酶。幸運的是,藥物感受性試驗中,imipenem或meropenem對於全部mcr-1陽性多重抗藥革蘭氏陰性菌皆成具有感受性,對於帶有mcr-1的質體做進一步分析發現質體屬於IncI2及IncX4,分別占43.8%及25%,但並沒有偵測到IncHI2的存在。

Bacteria with multiple drug resistance emerges globally and causes serious nosocomial infections. It is difficult to treat and control, and causes high mortality. Colistin is one of the last-resort antibiotic against most gram-negative bacteria especially carbapenem-resistance Enterobacteriaceae in treatment of human beings. In 2015, a plasmid-mediated colistin resistance gene (mcr-1) was reported for the first time in China. Also, it has been found in wildlife, humans and food over than 30 countries, including Taiwan.
Researches of plasmid-mediated colistin resistance gene from animals were few in Taiwan. This study aimed to investigate the prevalence of mcr-1 positive gram-negative isolates and overview of antibiotic resistance in clinical isolates from livestocks, wildlife and companion animals in Taiwan. Here we have screened colistin resistance gene in the clinical isolates from both the Animal Disease Diagnostic Center, College of Veterinary Medicine, and National Chung Hsing University from January 2016 to June 2017 and from National Chung Hsing University College of Veterinary Medicine from January 2016 to April 2017. We have collected 247 non-duplicate gram-negative clinical isolates. Screen all sample for plasmid-mediated colistin resistance gene (mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5). Our result showed that rate of colistin resistance from swine, poultry, wildlife and companion animal were 33% (42/126), 43% (20/47), 24.3% (9/34) and 8% (3/37) and prevalence rate of mcr-1 were 27.0% (34/126), 21.3% (10/47), 8% (3/37) and 2.7% (1/37), respectively. Also, mcr-3 was detected in 3 isolates from swine and 1 isolate from poultry. Coexistence of mcr-1 and mcr-3 (1%, three isolates) was observed from both swine and poultry. The mcr-2, mcr-4 and mcr-5 genes were not detected in any samples.
There were 48 gram-negative isolates with mcr-1, including 45 isolates of E. coli, 1 isolates of Aeromonas trota, Acinetobacter baumannii and Salmonella Enteritidis. Except A. trota, all mcr-1 positive isolates were resistant to colistin, ampicillin and tetracycline, and more than 90% resistance to trimethoprim-sulfamethoxazole and first generation cephalosporin. 35.4% mcr-1 positive isolates were ESBLs producer. Fortunately, we found that all mcr-1-positive multidrug-resistant gram-negative isolates were susceptible for imipenem or meropenem. We futher investigated type of mcr-1-carrying plasmid belonged to IncI2 and IncX4, respectively 43.8% and 25%. IncHI2 plasmid type was not detected in any isolate.
According to our result, this is first report to present of mcr-3 in Taiwan, and percentage of mcr-1 gene detected in E. coli from farm animals was high. The likely cause is that the usage of colistin for livestock is more often. Therefore, additional antibiotic stewardship program should keep monitoring.
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