EXECUTIVE SUMMARY Candida spp. especially Candida auris is a newly emerging human pathogenic multi-drug-resistant pathogen that causes nosocomial transmission between hospitals worldwide. Latest reports call attention to current challenges due to the misidentification of Candida spp. and high resistance to antimycotic drugs, and mortality of these organisms. This study was designed to investigate the status of Candida and Candidiasis among the immunocompromised patients admitted to different wards (Special Care and Intensive Care Units, Burn ward, Neuro-ICU, Special-Care Dialysis unit, and General Dialysis ward) of four different hospitals in Quetta City, Balochistan. Swab samples from the ear, axilla, groin, and oral samples of saliva/sputum were collected from a total of 500 patients (1600 samples) from Sandeman Provincial, Fatima Jinnah Chest Hospitals, Bolan Medical complex, Balochistan Institute of Nephro-Urology Quetta (SPHQ, FJCH, BMC & BINUQ) Quetta irrespective of their a

Summary of the Project: 1. Isolated human pathogenic Candida species associated from affected patients. 2. Cultured the associated pathogenic Candida species on culture media 3. Observed the various morphological features of colony and vegetative yeast cells using compound light microscope. 4. Identified the human pathogenic Candida species using Conventional DNA PCR technique and sequencing the Internal Transcribe Spacer (ITS) regions. 5. Designed the species-specific primers for targeted identification of human pathogenic Candida species using Real Time PCR Technique 6. Established clinical diagnostics of Candida species based on Real Time PCR Technique for direct benefits to affected patients.

Summary Postharvest diseases of fruits which are mainly caused by fungi cause severe losses. There are various means for protecting fruits from fungal attack. The use of environment friendly antagonistic yeasts are gaining particular attention. Several bio- fungicides based of yeast formulation are now available in market. In the present study, therefore, we have focused to isolate various fungal pathogens and environment friendly yeasts associated with the fresh fruits especially in the province of Balochistan. Investigating fresh fruits such as kinnow, apples, pomegranate and peach was found several pathogenic fungi including Aspergillus fumigatus, A. neoellipticus, A. tubingensis, A. albamensis, A. terreuis, Alternaria alternata, A. arbobrescens, A. brassicae, Corynespora cassiicola, , Alternaria porri, Lasiodiplodia theobromae, Chaetomiun sp., Penicilium dipodomyicola, P. granulatum, P. polonicum, P. adametzioides, P. bussumense, P. commune, P. spinulosum, Stemphylium solan

Summary: Postharvest fungal diseases cause significant losses to harvested fruits during transportation and storage. Fungi are ubiquitous in both natural and human-made environments. They play important roles in the health of plants, animals, and humans, and in broad ecosystem functions. The study of fungal pathogens has been happening from olden days to prevent epidemics, food spoilage, losses in agricultural production, and loss of lives. Thus, having an effective species-level identification system could considerably boost our ability to treat fungal diseases. DNA barcode is a very small, standardized DNA sequence in a familiar gene. It gives a way to identify the species to which a plant, animal or fungus belongs. DNA barcoding is a good concept of identifying unknown pathogenic fungi based on species-specific DNA regions. The nuclear ribosomal Internal Transcribed Spacer ITS region is widely used as a DNA barcoding marker to characterize the biodiversity and composition of funga

Summary In the present study, therefore, various fungal pathogens associated with fresh fruits such as kinnow, apples, pomegranate and peach in Balochistan province wer isolated using standard isolation techniques. Aspergillus fumigatus, A. neoellipticus, A. tubingensis, A. albamensis, A. terreuis, Alternaria alternata, A. arbobrescens, A. brassicae, Corynespora cassiicola, , Alternaria porri, Lasiodiplodia theobromae, Chaetomiun sp., Penicilium dipodomyicola, P. granulatum, P. polonicum, P. adametzioides, P. bussumense, P. commune, P. spinulosum, Stemphylium solan, Trichoderma lixi and Verticillium nubilum. All the isoalted pathogens were primarily identified by observing their macroscopic and microscopic features and finally by DNA sequencing of their PCR amplified products of ITS regions.

In the present, clinical specimens of urine were collected and analyzed for the presence of Candida species using CHROMagar technique, RapID Yeast Plus System, PCR-RFLP pattern and DNA sequencing. A total of 5 Candida species viz., C. albicans, C. glabrata, C. tropicalis, C. krusei and C. parapsilosis were identified, where occurrence of C. albicans was highest as compared to other species. Candida glabrata also recorded as an emerging fungal pathogen in fungiuria. It was observed that CHROMagar technique is reliable in routine laboratory testing but not in case of C. glabrata that produced conflicting colony colors. The RapID yeast Plus System found to be rapid but not much suitable for C. krusei and C. parapsilosis as it gives somewhat ambiguous results. PCR-RFLP and DNA sequencing of PCR products appeared to be the most reliable techniques for Candida species identification.

In this study we focused not only on the isolation and identification of pathogenic yeast species but also examined the frequency of candidiasis in different clinical specimens from affected patients. A total of 345 clinical specimens including urine, blood, sputum, vaginal swabs and pus were collected and cultured on YM agar culture medium for the isolation of associated pathogenic yeasts. This study describes the use of a simple and rapid method by PCR using universal primers ITS1 and ITS4 for the identification of clinical yeast isolates. Ninety (90) yeast strains from clinical samples were identified among which the most abundant species was Candida albicans. This species alone account 71.1% (64 isolates) as compared to C. glabrata (28.9%). The samples of blood, pus and sputum were 100% found to be affected by C. albicans; whereas the it was also found to be as a major causal agent in vaginal (74.3%) and urine (57.5%) infections as compared to C. glabrata.

Studies on killer and antagonistic activity of yeasts against plant pathogenic sclerotial fungi.

Identification and screening of yeasts showing killer and antagonistic activity against plant pathogenic fungi

Cultural and cytological studies on yeasts isolated from various ecological niches.

A survey of capsicum fields of Mirpur Khas to determine the possible cause(s) in yield reduction. Several plant pathogenic fungi were isolated from affected plants and soil. in vitro experiments were performed to check pathogenicity of certain plant pathogenic fungi and documented.