Multiple drug resistance in hookworms infecting greyhound dogs in the USA.
Multiple drug resistance in hookworms infecting greyhound dogs in the USA.
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DOI：
10.1016/j.ijpddr.2021.08.005
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发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Kaplan RM
中科院分区：
文献类型：
--
作者：
Jimenez Castro PD;Venkatesan A;Redman E;Chen R;Malatesta A;Huff H;Zuluaga Salazar DA;Avramenko R;Gilleard JS;Kaplan RM
犬钩虫是犬最常见的线虫寄生虫。我们证实了多个A.犬分离株与美国批准用于治疗犬钩虫的所有驱虫药类别一致。MDR钩虫病例在灰狗中的比例似乎过高。本研究的目的是评估耐药表型和基因型的A。感染灰狗的犬科动物美国灰狗的粪便样本来自两个灰狗收养狗舍、一个活跃的灰狗比赛狗舍和三个兽医诊所。对219只灰狗的粪便样本进行粪便虫卵计数（FEC），尽管用驱虫剂治疗，平均FEC为822.4个虫卵/克（EPG）。分别采用卵孵化试验（EHA）和幼虫发育试验（LDA）测定了对苯并咪唑类和大环内酯类的抗性。我们对代表54只动物的单个或合并粪便进行了23次EHA和22次LDA。EHA的平均和中位IC 50和IC 95值分别为5.3 μM、3.6 μM和24.5 μM、23.4 μM。对于LDA，中位IC 50值>1000 nM。这些值比我们的敏感实验室分离株高62-81倍。仅可获得治疗后样本。对于阿苯达唑、莫昔克丁或非班太尔-噻嘧啶-莫昔克丁联合给药后<10天采集的样本，平均FEC分别为349、333和835 EPG。我们从70个粪便样本中分离出钩虫卵，其中包括60只狗和10个池。同种型1 β-微管蛋白基因的深度测序仅揭示了99%的这些样品中存在F167 Y（TTC>TAC）抗性多态性。这些临床、体外和遗传数据提供了强有力的证据，证明美国的灰狗感染了MDR A。犬在患病率和感染强度非常高的水平。结论性证据表明，大多数赛狗在美国感染了MDR A。犬的79%的赛狗或退役赛狗样本呈阳性。BZ和ML的IC 50值在灰狗中高62-81倍。在99%的样本中检测到F167 Y SNP，并且在超过2/3的样本中频率≥ 75%。
Ancylostoma caninum is the most prevalent nematode parasite of dogs. We confirmed multiple-drug resistance (MDR) in several A. caninum isolates to all anthelmintic drug classes approved for the treatment of hookworms in dogs in the USA. Cases of MDR hookworms appear to be highly overrepresented in greyhounds. The aims of this study were to evaluate the drug-resistant phenotypes and genotypes of the A. caninum infecting greyhounds. Fecal samples from greyhounds of the USA were acquired from two greyhound adoption kennels, one active greyhound racing kennel, and three veterinary practices. Fecal egg counts (FECs) were performed on fecal samples from 219 greyhounds, and despite treatment with anthelmintics, the mean FEC was 822.4 eggs per gram (EPG). Resistance to benzimidazoles and macrocyclic lactones were measured using the egg hatch assay (EHA) and the larval development assay (LDA), respectively. We performed 23 EHA and 22 LDA on either individual or pooled feces, representing 54 animals. Mean and median IC50 and IC95 values for the EHA were 5.3 μM, 3.6 μM, and 24.5 μM, 23.4 μM, respectively. For the LDA, the median IC50 value was >1000 nM. These values ranged 62–81 times higher than our susceptible laboratory isolate. Only post-treatment samples were available. For samples collected <10 days post-treatment with albendazole, moxidectin, or a combination of febantel-pyrantel-moxidectin, the mean FEC were 349, 333, and 835 EPG, respectively. We obtained DNA from hookworm eggs isolated from 70 fecal samples, comprised of 60 individual dogs and 10 pools. Deep sequencing of the isotype 1 β-tubulin gene only revealed the presence of the F167Y (TTC>TAC) resistance polymorphism in 99% of these samples. These clinical, in vitro, and genetic data provide strong evidence that greyhound dogs in the USA are infected with MDR A. caninum at very high levels in prevalence and infection intensity. Conclusive evidence that most racing greyhounds in the USA are infected with MDR A. caninum. 79% of the samples from racing or retired greyhounds were positive. IC50 values for BZs and MLs were 62–81 times higher in greyhounds. The F167Y SNP was detected in 99% of samples, and in more than 2/3 the frequency was ≥ 75%.
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