鉴别显色培养基在奶牛乳房炎病原菌快速鉴定中的应用

doi:10.11843/j.issn.0366-6964.2019.04.018

Abstract

Abstract:

The present study aimed to develop a new differential chromogenic medium for fast identification of pathogens associated with mastitis in dairy cows, and check its identification consistency with two traditional approaches including normal isolation and identification media combined with biochemical identifications and 16S rRNA sequencing. Specific enzymes and the only carbon source that could be used to produce acid in bacteria of each genus were screened via bioinformatics tools. Synthesized chromogenic substrate of specific enzyme, carbon and acid-base indicator were mixed into basal enrichment medium to prepare differential chromogenic media. Standard strains and wild-type strains of related pathogens were inoculated on differential chromogenic media, and morphology of colonies and color changes of colony and medium that could be considered as identification standards were observed. Milk samples from mastitic quarters (samples diagnosed as subclinical mastitis by California Mastitis Test or samples of clinical mastitis with presence of flakes, clots, or serous milk; n=482) were cultured aerobically at 37℃ and identified using differential chromogenic media and traditional culture methods. Colonies from differential chromogenic media were purified and DNA was extracted for 16S rRNA sequencing (n=194). The reliability of identification by differential chromogenic media was diagnosed referring to results of two traditional approaches, based on which consistency inspection parameter, namely, simple Cohen's kappa coefficient was calculated via FREQ program in SAS Version 9.4. The colony and color of medium matrix from common bovine mastitis-associated pathogens of different genus grown on differential chromogenic media, regardless of standard strains or wild-type strains, were morphologically distinct and visible to the naked eyes. Consistency inspection parameter of differential chromogenic media referred to normal isolation and identification media combined with biochemical identifications and 16S rRNA sequencing was seperately κ=0.70 and κ=0.96. In conclusion, differential chromogenic media showed great potential in identifying common pathogens of bovine mastitis.

Key words: dairy cow, mastitis, fast identification of pathogens, differential chromogenic medium

CLC Number:

S852.61

Lü Tianxing, LI Songjian, HAO Yongqing. Application of a New On-farm Differential Chromogenic Medium for Fast Identification of Pathogens Associated with Mastitis in Milk of Dairy Cows[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 851-860.

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Application of a New On-farm Differential Chromogenic Medium for Fast Identification of Pathogens Associated with Mastitis in Milk of Dairy Cows

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