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Short Communication Volume 11 Issue 2
First report of a yeast pathogen Diutina rugosa (syn. Candida rugosa) in an outbreak of mastitis in Holstein Friesian dairy heard in India
Amit Kumar,
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Anamika Bhordia, Surendra Upadhyay, Malyaj R Prajapati, Jitender Singh, Ravindra Kumar
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Division of Animal Biotechnology, College of Biotechnology, Sardar Vallabhbhai Patel University of Agriculture & Technology (SVPUAT), India
Correspondence: Amit Kumar, Division of Animal Biotechnology, College of Biotechnology, Sardar Vallabhbhai Patel University of Agriculture & Technology (SVPUAT), Meerut-250110, India, Tel +91-9412120813
Received: March 03, 2023 | Published: March 21, 2023
Citation: Bhordia A, Kumar A, Upadhyay S, et al. First report of a yeast pathogen Diutina rugosa (syn. Candida rugosa) in an outbreak of mastitis in Holstein Friesian dairy heard in India. J Microbiol ExpM. 2023;11(2):43-44. DOI: 10.15406/jmen.2023.11.00384
Short Communication
Mastitis is one of the most prevalent causes of economic losses to dairy sector.1 It involves multiple etiological agents and that makes it difficult to cure.2,3 It is therefore, important to isolate, identify the causing agent for its earliest best effective treatment to avoid or to minimize losses.1 Accordingly, when a dairy, with 30 pure bred Holstein Friesian cattle, was reported with an outbreak of mastitis, the samples were received from local veterinarian to suggest the best possible antibiotic treatment but failed to control mastitis. The milk samples were submitted to than Department of Immunology & Defense Mechanism, College of Biotechnology, SVPUAT, Meerut for the isolation of bacterial pathogens and also to suggest suitable antibiotic treatment based on antibiotic sensitivity assay. None of the sample revealed bacterial growth. The fresh milk samples were collected from the untreated animals and cultured on different culture media including Potato Dextrose Agar (PDA).4 After 24 hours of incubation, PDA revealed some creamish white, round, smooth and raised colonies over the plate. Figure 1.
Figure 1 Creamy to whitish colonies of Diutina rugosa (syn. Candida rugosa).
The staining of these colonies revealed the yeast like appearance on Gram staining. Figure 2
Figure 2 Presence of budding cells suggesting the presence of yeast.
As the monoculture was obtained, it was subjected to DNA extraction and amplification of 26sRNA using ITS primer.5 The PCR revealed the presence of 500bp amplicons. Figure 3
Figure 3 ThreeITS primers based PCR revealed 500bp amplicons.
The PCR products were outsourced to Biokart Pvt Ltd (India) for senger sequencing. The processing and BLAST of obtained sequences revealed Diutina rugosa (syn. Candida rugosa). The sequenced data has been submitted to NCBI database and assigned an accession number MT936353. The study for phylogeny linkage revealed Indian isolate of Diutina rugosa in close homology with the isolate of Thailand and both share a same route followed by the isolates of China, Ghana and Mexico. It reveals the ancestral linkage with Indian and Thai isolates. Figure 4
Figure 4 Phylogenetic tree showing the evolutionary relationship of the isolated species Diutina rugosa (marked with ) with the same species isolated from different countries.
Diutina rugosa (syn. Candida rugosa) are considered as “rare yeasts” and has been reported from different clinical conditions of animals. The drug susceptibility of isolates revealed susceptibility to azole compounds like fluconazole and less or no susceptibility to amphotericin B. Further, it is considered as an emerging fungal pathogen. A 10-year multi-center study reported 10 fold increase in the number of D. rugosa clinical cases.6 However, there is no such report of its involvement in lactating animals in India in available literature. Thus, it seems first report of an outbreak of mastitis due to Diutina rugosa (syn. Candida rugosa) from Indian subcontinent.
Ethical Approval: The ethical approval for the present study was obtained vide letter no. IAEC/ SVPUAT/2019/18 dated 25/07/2019 and CPCSEA, New Delhi vide letter no. V-11011(13)/2/2020 CPCSEA-DAFA dated 10/02/2020.
Conflicts of interest
Authors declare no conflict of interest.
Acknowledgments
Authors are thankful the Director Research and Hon’ble Vice Chancellor, SVPUAT, Meerut for providing all the facilities required to conduct the study.
References
- Kumar A, Rahal A, Dwivedi SK. Prevalence and antibiotic profile of bacterial isolates from bovine mastitis in Mathura. Egyptian J Dairy Sci. 2010;38(1):31‒34.
- Kumar A, Verma AK, Rahal A. Mycoplasma bovis, A multi disease producing pathogen: an overview. Asian J Anim Vet. Advances. 2011;6(6):537‒546.
- Kumar A, Verma AK, Gupta MK, et al. Outbreak of mastitis in sheep flock due to Streptococcus agalactiae and unusual neonatal lamb mortality. Adv Anim Vet Sci. 2014;1(4):120‒122.
- Bergey DH, Holt JG, Krieg P. Bergey's manual of determinative bacteriology. Williams and Wilkins, Baltimore, MD, USA; 1994.
- Kwiatkowski NP, Babiker WM, Merz WG, et al. Evaluation of nucleic acid sequencing of the D1/D2 region of the large subunit of the 28S rDNA and the internal transcribed spacer region using smart gene idn software for identification of filamentous fungi in a clinical laboratory. J Mol Diagnostics. 2012;14:393–401.
- Pfaller MA, Diekema DJ, Gibbs DL, et al. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: a 10.5-year analysis of susceptibilities of Candida Species to fluconazole and voriconazole as determined by CLSI standardized disk diffusion. J Clin Microbiol. 2010;48:1366–1377.
©2023 Bhordia, et al. This is an open access article distributed under the terms of the, which permits unrestricted use, distribution, and build upon your work non-commercially.
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