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. 2018 Aug 29;14(1):251.
doi: 10.1186/s12917-018-1564-3.

Epidemiological investigation of non-albicans Candida species recovered from mycotic mastitis of cows in Yinchuan, Ningxia of China

Jun Du  1   2 , Xiaoyu Wang  1   2 , Huixia Luo  1   2 , Yujiong Wang  3   4 , Xiaoming Liu  5   6 , Xuezhang Zhou  7   8
Affiliations

Epidemiological investigation of non-albicans Candida species recovered from mycotic mastitis of cows in Yinchuan, Ningxia of China

Jun Du et al. BMC Vet Res. .

Abstract

Background: Candida spp. is the vital pathogen involved in mycotic mastitis of cows. However the epidemiology and infection of Candida species in mycotic mastitis of cow in Ningxia province of China has not been explored. In the present study, the epidemiology, antimicrobial susceptibility and virulence-related genes of non-albicans Candida (NAC) species were investigated.

Methods: A total of 482 milk samples from cows with clinical mastitis in four herds of Yinchuan, Ningxia were collected and used for the isolation and identification of mastic pathogens by phenotypic and molecular characteristics, and matrix-assisted laser desorption ionization-time of flight mass spectrometry. The antimicrobial susceptibility to antifungal agents was also determined by a disk diffusion assay. The presence of virulence-related genes was determined by polymerase chain reaction (PCR).

Results: A total of 60 isolates from nine different Candida species were identified from 256 (60/256, 23.44%) milk samples. The most frequently identified species in cows with clinical mastitis groups were Candida krusei (n = 14) and Candida parapsilosis (n = 6). Others include Candida lipolytica, Candida lusitaniae, Cryptococcus neoformans. But no Candida albicans was identified in this study. Interestingly, All C. krusei isolates (14/14) were resistant to fluconazole, fluorocytosine, itraconazole and ketoconazole, 2 out of 14 C. krusei were resistant to amphotericin, and 8 out of the 14 were resistant to nystatin. Similarly, all six C. parapsilosis isolates were resistant to fluorocytosine, but susceptible to fluconazole, ketoconazole and nystatin; two of the six were resistant amphotericin and itraconazole. Molecularly, all of the C. parapsilosis isolates carried eight virulence-related genes, FKS1, FKS2, FKS3, SAP1, SAP2, CDR1, ERG11 and MDR1. All of the C. krusei isolates contained three virulence-related genes, ERG11, ABC2 and FKS1.

Conclusion: These data suggested that Candida species other than C. albicans played a pathogenic role in mycotic mastitis of cows in Yinchuan, Ningxia of China. The high incidence of drug-resistant genes in C. parapsilosis and C. krusei also highlighted a great concern in public and animal health in this region.

Keywords: Antimicrobial susceptibility; Candida krusei; Candida parapsilosis; Mycotic mastitis; Virulent gene.

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Conflict of interest statement

Ethics approval and consent to participate

This study was conducted in accordance with the Law on Animal Protection and Welfare of Ningxia Hui Autonomous Region of China. Samples were recovered after acquiring the permission from study participants. Since only the milk samples but not mammary tissues were harvested in this study, all owners of farmers who participated in this provided their verbal consent to participate in this study. This study was submitted to and approved by the Ethics Committee of Animal Study in Ningxia University.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Representative culture and Morphology of C. krusei and C. parapsilosis isolates. a Representative culture of C. krusei on CHROMagar plate; a’ the morphology of Giemsa staining for yeast and pseudohyphae of a C. krusei isolate in a; b Representative culture of C. parapsilosis on CHROMagar plate; b’ the morphology of Giemsa staining for yeast and pseudohyphae of a C. parapsilosis isolate in (b)
Fig. 2
Fig. 2
Representative antimicrobial susceptibility tests by disk diffusion assay for C. krusei and C. parapsilosis isolates. a Representative disk diffusion assay for C. krusei isolate cultured on CHROMagar plate with indicated antimicrobial disks; b Representative disk diffusion assay for a C. parapsilosis isolate cultured on CHROMagar plate with indicated antimicrobial disks. Am, amphotericin; Fs, fluorocytosine; Fz, fluconazole; It, itraconazole; Ke, ketoconazole; Ny, nystatin
Fig. 3
Fig. 3
Virulence-related genes of C. krusei isolates determined by PCR assay. The indicated virulence-related genes of (a) FSK1, b ABC2 and (c) EGR11 in 14 field C. krusei isolates of this report were detected by PCR assay. C, control C. krusei ATCC6258 strain; lanes 1 to 14 represented C. krusei isolate 1–14. M, 50 bp DNA ladders
Fig. 4
Fig. 4
Virulence-related genes of C. parapsilosis isolates determined by PCR assay. The indicated virulence-related genes of (a) FSK1, b FSK2, c FSK3 (d) SAPP1, e SAPP2, f CDR1, g MDR1 and (h) EGR11 in 6 field C. parapsilosis isolates of this report were detected by PCR assay. c, C. parapsilosis ATCC22019 control strain; lanes 1 to 6 represented C. parapsilosis isolate 1–6. M, 2 kb DNA ladders
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