Isolation, Identification and Antibiotics Resistance of <i>Corynebacterium pseudotuberculosis</i> from Goats

HUANG Li-li; LIN Yu-sheng; JIANG Jin-xiu; ZHANG Jing-peng; YOU Wei; XIAO Yi-jun; HU Qi-lin

doi:10.19303/j.issn.1008-0384.2019.08.009

Volume 34 Issue 8

Aug. 2019

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Fujian Journal of Agricultural Sciences > 2019 > 34(8): 925-932. > DOI: 10.19303/j.issn.1008-0384.2019.08.009

HUANG Li-li, LIN Yu-sheng, JIANG Jin-xiu, ZHANG Jing-peng, YOU Wei, XIAO Yi-jun, HU Qi-lin. Isolation, Identification and Antibiotics Resistance of Corynebacterium pseudotuberculosis from Goats[J]. Fujian Journal of Agricultural Sciences, 2019, 34(8): 925-932. DOI: 10.19303/j.issn.1008-0384.2019.08.009

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Isolation, Identification and Antibiotics Resistance of Corynebacterium pseudotuberculosis from Goats

1.
College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, China
2.
Institute of Animal Husbandry and Veterinary, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China

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Received Date: May 27, 2019
Revised Date: August 04, 2019

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Abstract

Abstract

Objective To identify the pathogen that caused the death of goats in a farm in Fujian for prevention and treatment of Pseudotuberculosis.
Method Aseptically collected specimens of lung tissues from the dead goats were used to isolate the microbes for morphological observation and strain identification. DNA of the isolates were extracted, and the specific and 16S rRNA universal primers selected for PCR. The 16S rRNA amplified fragment was cloned, sequenced and analyzed. Biochemical, drug sensitivity, and animal challenge tests were performed on the isolate for taxonomic identification.
Results The isolated strain was cultured on an agar medium containing 10% defibrated sheep blood to grow into a colony with a diameter of about 1 mm, a central bulge, a milky white color, an irregular edge, and a beta hemolysis ring. It did not grow well on a nutrient agar nor a MacConkey agar medium. The 16S rRNA sequencing showed an up to 100% homology with Corynebacterium pseudotuberculosis on GenBank by the NCBI online software Blast. Like the specific PCR of C.pseudotuberculosis, that of the isolate also had the same amplified targeted bands. In addition, the biochemical test on the isolate duplicated the characteristics of C. pseudotuberculosis shown in the Berger's Bacterial Identification Manual. Hence, the isolated pathogen named as FJ-PN was verified to be C. pseudotuberculosis. A challenge test on mice with an intraperitoneal or subcutaneous injection of 0.3 mL of 4×10⁵ CFU·mL^-1 of the isolate died within 40 h. Furthermore, two goats injected subcutaneously with 2 mL of 4×10⁵ CFU·mL^-1 and intranasal inoculated simultaneously with 1 mL of 4×10⁵ CFU·mL^-1 of the isolate died within 72 h. The drug sensitivity test showed that the strain was highly sensitive to clarithromycin, ciprofloxacin, penicillin, tetracycline, levofloxacin, ofloxacin, and gentamicin, slightly sensitive to erythromycin, chloramphenicol, ampicillin, cefotaxime, cefoxitin, and rifampicin, and resistant to streptomycin or trimethoprim.
Conclusion The pathogenic bacteria isolated from the diseased goats was identified as C. pseudotuberculosis. It had a strong pathogenicity with varying degrees of drug resistance to antibiotics.
- Corynebacterium pseudotuberculosis,
- goat,
- antibiotics resistance,
- isolation,
- identification

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Isolation, Identification and Antibiotics Resistance of Corynebacterium pseudotuberculosis from Goats

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