Isolation and Molecular Analysis of Cryptococcus laurentii from Pigeon Feces in Urban and Suburban Regions

Abstract

It was reported that invasive fungal infections result in significant morbidity and mortality rates, particularly in immunocompromised hosts. Although pigeon droppings might be a possible vector for the transmission of pathogenic yeasts, The goal of this study was to isolate and identify Cryptococcus species using genetic and phenotypic methods. The results were then confirmed by doing several tests on pigeon droppings. In this research paper, two hundred samples of pigeon droppings were collected from fifty-three diverse places in Erbil city and its suburbs between November 2021 and October 2022. It has been experimentally found that among these two hundred samples, a total of fifteen isolates of Cryptococcus were obtained from pigeon droppings in the aforementioned places. In addition, Cryptococcus laurentii was identified by culturing on Sabouraud Dextrose Agar (SDA) and Niger Bird Seed Agar (NBSA) based on capsule formation, biochemical methods using VITEK, Urease test, and Phenol oxidase test, and then confirmed by a molecular technique such as DNA extraction and PCR amplification of ITS region sequence by using universal primers. The sequence data of C. laurentii accession number of nucleotide OR509919 have been provided by GenBank available on the NCBI website. In the biofilm formation method, C. laurentii isolates produced biofilms according to both phenotypic and molecular methods in the biofilm formation method, C. laurentii isolates produced biofilms, according to both phenotypic using Congo Red Agar media (CRA), Microtiter Plate Approach (MTP) and molecular methods for amplification of the ALS1 and HWP1 genes. It has been concluded that the utilization of the multiplex-PCR technique is highly suggested as a technology that offers enhanced detection of Cryptococcus spp. with high sensitivity and specificity.