Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP

Mastitis is caused by various microorganisms like bacteria, viruses, mycoplasma, algae and yeasts. Of them, fungal infections contribute to 2–13% of the cases, with Candida as the most common genus, although wide variations in prevalence and species are identified. In the present study, 06.66% (n =...

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Main Authors: DEVENDER SINGH RANA, PANKAJ KUMAR, PAWAN KUMAR, JASLEEN KAUR, SAURABH J TALUKDAR, RAHUL YADAV, MAHAVIR SINGH, JINU MANOJ, RAJESH CHHABRA
Format: Article
Language:English
Published: Indian Council of Agricultural Research 2025-07-01
Series:Indian Journal of Animal Sciences
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Online Access:https://epubs.icar.org.in/index.php/IJAnS/article/view/156881
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author DEVENDER SINGH RANA
PANKAJ KUMAR
PAWAN KUMAR
JASLEEN KAUR
SAURABH J TALUKDAR
RAHUL YADAV
MAHAVIR SINGH
JINU MANOJ
RAJESH CHHABRA
author_facet DEVENDER SINGH RANA
PANKAJ KUMAR
PAWAN KUMAR
JASLEEN KAUR
SAURABH J TALUKDAR
RAHUL YADAV
MAHAVIR SINGH
JINU MANOJ
RAJESH CHHABRA
author_sort DEVENDER SINGH RANA
collection DOAJ
description Mastitis is caused by various microorganisms like bacteria, viruses, mycoplasma, algae and yeasts. Of them, fungal infections contribute to 2–13% of the cases, with Candida as the most common genus, although wide variations in prevalence and species are identified. In the present study, 06.66% (n = 40/600) prevalence was observed in mastitic milk samples of cattle and buffalo on the basis of culture examination on blood agar (BA), sabouraud dextrose agar (SDA) and candida differentiation agar (CDA). On SDA, the isolates produced white to creamy, smooth, pasty and convex colonies. However, different isolates could not be differentiated on the basis of colony characteristic on SDA medium. Chromogenic agar such as candida differential agar was able to differentiate different species of Candida on the basis of production of different coloured colonies. In the VITEK®2 compact system, different isolates were identified with diverse level of identification ranging from excellent to acceptable. Furthermore, all of the isolates were confirmed by amplification of internal transcribed spacer (ITS) region by polymerase chain reaction (PCR). For molecular characterization of Candida spp., the PCR products were digested by restriction enzymes (HaeIII and TaqI) for restriction fragment length polymorphism (RFLP) and various patterns were obtained. Furthermore, in-silico RFLP analysis was done with restriction enzymes HaeIII and TaqI, for different Candida species to obtain accurate fragment sizes. PCR-RFLP proved to be a simple, cost effective and accurate method as compared to the phenotypic based methods which are often difficult and time consuming for rapid and species level differentiation of Candida species involved in mycotic mastitis.
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spelling doaj-art-89e01324d8904716942e43030db28fa12025-08-21T12:25:48ZengIndian Council of Agricultural ResearchIndian Journal of Animal Sciences0367-83182394-33272025-07-0195410.56093/ijans.v95i4.156881Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLPDEVENDER SINGH RANA0PANKAJ KUMAR1PAWAN KUMAR2JASLEEN KAUR3SAURABH J TALUKDAR4RAHUL YADAV5MAHAVIR SINGH6JINU MANOJ7RAJESH CHHABRA8Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, HaryanaLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana Mastitis is caused by various microorganisms like bacteria, viruses, mycoplasma, algae and yeasts. Of them, fungal infections contribute to 2–13% of the cases, with Candida as the most common genus, although wide variations in prevalence and species are identified. In the present study, 06.66% (n = 40/600) prevalence was observed in mastitic milk samples of cattle and buffalo on the basis of culture examination on blood agar (BA), sabouraud dextrose agar (SDA) and candida differentiation agar (CDA). On SDA, the isolates produced white to creamy, smooth, pasty and convex colonies. However, different isolates could not be differentiated on the basis of colony characteristic on SDA medium. Chromogenic agar such as candida differential agar was able to differentiate different species of Candida on the basis of production of different coloured colonies. In the VITEK®2 compact system, different isolates were identified with diverse level of identification ranging from excellent to acceptable. Furthermore, all of the isolates were confirmed by amplification of internal transcribed spacer (ITS) region by polymerase chain reaction (PCR). For molecular characterization of Candida spp., the PCR products were digested by restriction enzymes (HaeIII and TaqI) for restriction fragment length polymorphism (RFLP) and various patterns were obtained. Furthermore, in-silico RFLP analysis was done with restriction enzymes HaeIII and TaqI, for different Candida species to obtain accurate fragment sizes. PCR-RFLP proved to be a simple, cost effective and accurate method as compared to the phenotypic based methods which are often difficult and time consuming for rapid and species level differentiation of Candida species involved in mycotic mastitis. https://epubs.icar.org.in/index.php/IJAnS/article/view/156881Buffalo, Candida spp., Cattle, RFLP, Mastitis
spellingShingle DEVENDER SINGH RANA
PANKAJ KUMAR
PAWAN KUMAR
JASLEEN KAUR
SAURABH J TALUKDAR
RAHUL YADAV
MAHAVIR SINGH
JINU MANOJ
RAJESH CHHABRA
Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
Indian Journal of Animal Sciences
Buffalo, Candida spp., Cattle, RFLP, Mastitis
title Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
title_full Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
title_fullStr Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
title_full_unstemmed Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
title_short Phenotypic and molecular characterization of Candida spp. isolated from intramammary infections in dairy animals by PCR-RFLP
title_sort phenotypic and molecular characterization of candida spp isolated from intramammary infections in dairy animals by pcr rflp
topic Buffalo, Candida spp., Cattle, RFLP, Mastitis
url https://epubs.icar.org.in/index.php/IJAnS/article/view/156881
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