{"title":"Molecular detection and genetic analysis of <i>Candida</i> species isolated from bovine clinical mastitis in India.","authors":"Nivedha Devanathan, Mouttou Vivek Srinivas, Jayalakshmi Vasu, Shashank Chaluva, Hirak Kumar Mukhopadhyay","doi":"10.30466/vrf.2024.2008702.3968","DOIUrl":null,"url":null,"abstract":"<p><p>Bovine clinical mastitis is an economically important disease in dairy industry worldwide resulting in reduction of milk yield and quality. Among mycotic mastitis, <i>Candida</i> spp. are commonly occurring opportunistic mycosis in immunocompromised animals. The micro-organism's causing mastitis has high zoonotic potential and has been linked with rapid growth and introduction of antimicrobial resistance between animals and humans. The present study was conducted to isolate and identify the common pathogenic <i>Candida</i> spp. from bovine mastitis cases in India. The isolates were phenotypically characterized by culturing on Sabouraud's dextrose agar, Hichrome <i>Candida</i> differential agar and germ tube production test. Antibiogram was also performed to determine their antifungal activities. The phenotypically positive isolates were confirmed by polymerase chain reaction (PCR) and genetically analyzed by targeting 18S-ITS1-5.8S-ITS2-28S region specific for <i>Candida</i> spp. and identified the yeast at the species level. The antibiogram showed the isolates were highly sensitive with ketoconazole, clotrimazole and miconazole. The PCR assay identified <i>C. lusitaniae</i> and <i>C. tropicalis</i> based on the two distinctive amplicon sizes (592bp and 737bp) respectively. Also, the sequence analysis and phylogeny confirmed <i>C. lusitaniae</i> in six sequences and <i>C. tropicalis</i> in one sequence. It is worth noting that in this study, the species identification was consistent among PCR and genetic analysis. Therefore, the PCR based identification system of the fungal species performed in this study could be an efficient and time saving tool for early diagnosis of clinical mastitis in milch animal, which allows prompt control and application of speedy effective treatment.</p>","PeriodicalId":23989,"journal":{"name":"Veterinary Research Forum","volume":"15 9","pages":"509-514"},"PeriodicalIF":0.8000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11571044/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary Research Forum","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.30466/vrf.2024.2008702.3968","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/9/15 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"ZOOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Bovine clinical mastitis is an economically important disease in dairy industry worldwide resulting in reduction of milk yield and quality. Among mycotic mastitis, Candida spp. are commonly occurring opportunistic mycosis in immunocompromised animals. The micro-organism's causing mastitis has high zoonotic potential and has been linked with rapid growth and introduction of antimicrobial resistance between animals and humans. The present study was conducted to isolate and identify the common pathogenic Candida spp. from bovine mastitis cases in India. The isolates were phenotypically characterized by culturing on Sabouraud's dextrose agar, Hichrome Candida differential agar and germ tube production test. Antibiogram was also performed to determine their antifungal activities. The phenotypically positive isolates were confirmed by polymerase chain reaction (PCR) and genetically analyzed by targeting 18S-ITS1-5.8S-ITS2-28S region specific for Candida spp. and identified the yeast at the species level. The antibiogram showed the isolates were highly sensitive with ketoconazole, clotrimazole and miconazole. The PCR assay identified C. lusitaniae and C. tropicalis based on the two distinctive amplicon sizes (592bp and 737bp) respectively. Also, the sequence analysis and phylogeny confirmed C. lusitaniae in six sequences and C. tropicalis in one sequence. It is worth noting that in this study, the species identification was consistent among PCR and genetic analysis. Therefore, the PCR based identification system of the fungal species performed in this study could be an efficient and time saving tool for early diagnosis of clinical mastitis in milch animal, which allows prompt control and application of speedy effective treatment.
牛临床乳腺炎是全球乳制品行业的一种重要经济疾病，会导致牛奶产量和质量下降。在霉菌性乳腺炎中，念珠菌属是免疫力低下动物常患的机会性霉菌病。引起乳腺炎的微生物具有很高的人畜共患潜力，并且与动物和人类之间抗菌素耐药性的快速增长和引入有关。本研究旨在从印度的牛乳腺炎病例中分离和鉴定常见的致病性念珠菌属。通过在沙保鲁葡萄糖琼脂、Hichrome 念珠菌差异琼脂和菌管生产试验上进行培养，对分离物进行表型鉴定。还进行了抗菌谱分析，以确定它们的抗真菌活性。通过聚合酶链式反应（PCR）确认了表型阳性的分离物，并通过针对念珠菌属特异的 18S-ITS1-5.8S-ITS2-28S 区域进行基因分析，确定了酵母菌的种类。抗生素图谱显示，分离物对酮康唑、克霉唑和咪康唑高度敏感。根据两个不同的扩增子大小（592bp 和 737bp），PCR 检测确定了 C. lusitaniae 和 C. tropicalis。此外，序列分析和系统发育证实了六个序列中的 C. lusitaniae 和一个序列中的 C. tropicalis。值得注意的是，在本研究中，PCR 和基因分析的物种鉴定结果是一致的。因此，本研究中基于 PCR 的真菌物种鉴定系统可作为一种高效、省时的工具，用于乳畜临床乳腺炎的早期诊断，以便及时控制和应用快速有效的治疗。
期刊介绍：
Veterinary Research Forum (VRF) is a quarterly international journal committed to publish worldwide contributions on all aspects of veterinary science and medicine, including anatomy and histology, physiology and pharmacology, anatomic and clinical pathology, parasitology, microbiology, immunology and epidemiology, food hygiene, poultry science, fish and aquaculture, anesthesia and surgery, large and small animal internal medicine, large and small animal reproduction, biotechnology and diagnostic imaging of domestic, companion and farm animals.
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