Virulence Factors and Antimicrobial Resistance of Uropathogenic Escherichia coli EQ101 UPEC Isolated from UTI Patient in Quetta, Balochistan, Pakistan.

Infectious diseases have been tremendously increasing as the organisms of even normal flora become opportunistic and cause an infection, and Escherichia coli (E. coli EQ101) is one of them. Urinary tract infections are caused by various microorganisms, but Escherichia coli is the primary cause of almost 70%-90% of all UTIs. It has multiple strains, possessing diverse virulence factors, contributing to its pathogenicity. Furthermore, these virulent strains also can cause overlapping pathogenesis by sharing resistance and virulence factors among each other. The current study is aimed at analyzing the genetic variants associated with multi-drug-resistant (MDR) E. coli using the whole genome sequencing platform. The study includes 100 uropathogenic Escherichia coli (UPEC) microorganisms obtained from urine samples out of which 44% were multi-drug-resistant (MDR) E. coli. Bacteria have been isolated and antimicrobial susceptibility test (AST) was determined by disk diffusion method on the Mueller-Hinton agar plate as recommended by the Clinical and Laboratory Standards Institute (CLSI) 2020, and one isolate has been selected which shows resistance to most of the antibiotics, and that isolate has been analyzed by whole genome sequencing (WGS), accompanied by data and phylogenetic analysis, respectively. Organisms were showing resistance against ampicillin (10 µg), cefixime (5 µg), ceftriaxone (30 µg), nalidixic acid (30 µg), ciprofloxacin (5 µg), and ofloxacin (5 µg) on antimicrobial susceptibility test. WGS were done on selected isolate which identified 25 virulence genes (air, astA, chuA, fyuA, gad, hra, iha, irp2, iss, iucC, iutA, kpsE, kpsMII_K1, lpfA, mchF, ompT, papA_F43, sat, senB, sitA, terC, traT, usp, vat, and yfcV) and seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Among resistance genes, seven genes (TolC, emrR, evgA, qacEdelta1, H-NS, cpxA, and mdtM) were identified to be involved in antibiotic efflux, three AMR genes (aadA5, mphA, and CTX-M-15) were involved in antibiotic inactivation, and two genes (sul1 and dfrA14) were found to be involved in antibiotic drug replacement. Our data identified antibiotic resistance and virulence genes of the isolate. We suggest further research work to establish region-based resistance profile in comparison with the global resistance pattern.

Hospital Acquired Pathogenic Escherichia coli from Clinical and Hospital Water Samples of Quetta Balochistan.

A study was conducted to determine the prevalence and drug resistance of Escherichia coli present in urinary tract infected patients and hospital drinking water. A total of eighty urine samples from clinically suspected patients and thirty tap water samples from hospital vicinity were collected and analyzed for the presence of E. coli. The isolates were preliminary identified based on morphological characteristics, biochemical test and further confirmed by polymerase chain reaction (PCR) using uidA primer. Isolates were subjected to antibiogram studies and analyzed for the presence of drug resistance (ESBL blaCTX-M-15, tetA, and TMP-SMX dfrA1) and pathogenicity associated pyelonephritis-associated pili (PAP) and Heat-labile (LT) toxin genes. Urine samples 19/80 (23.75%) and water samples 8/30 (26.7%) were found contaminated with E. coli. It was found that 12/19 (63%) bacterial isolates were extended spectrum beta-lactamase (ESBL) producers in clinical and 6/8 (75%) in water isolates whereas tetracycline resistance in clinical and water isolates was 11/19 (58%) and 6/8 (75%), respectively. The trimethoprim resistance gene was confirmed in 12/19 (63%) in clinical and 2/8 (25%) in water isolates. All the clinical and water isolates were found carrying pili PAP gene. It was concluded that the presence of drug resistant and pathogenic E. coli in clinical and water samples is extremely alarming for public health due to cross contamination and bacterial transfer from clinical samples to water and vice versa.

Predominance of Central Asian strain (ST 26) in Mycobacterium tuberculosis isolates from Balochistan by spoligotyping.

INTRODUCTION: Tuberculosis is a chronic debilitating infectious disease causing a severe challenge to public health, especially in developing countries. The aim of this study was to examine genetic diversity in Mycobacterium tuberculosis strains circulating in the Balochistan region of Pakistan. METHODOLOGY: One hundred isolates collected from patients visiting the Fatima Jinnah TB Hospital in Quetta were subjected to genotype analysis by spoligotyping. RESULTS: Three main genotypes were identified: Central Asian Strain 1 (CAS1) (n = 89), East African Indian (EAI) strain (n = 7) and Latin American Mediterranean (LAM) strain (n = 3). The CAS1 clade (ST 26) had high genetic diversity represented by seven different spoligopatterns, of which one had major predominace (n = 75). CONCLUSIONS: This is the first insight into the genotype of M. tuberculosis strains in the Balochistan region that might serve as a base line study for control of tuberculosis in the community.

Hematological profile and risk factors associated with pulmonary tuberculosis patients in Quetta, Pakistan.

OBJECTIVES: Tuberculosis (TB) is a chronic debilitating infectious disease affecting more than one third of the global population. This study was designed to investigate different peripheral blood parameters and risk factors in TB patients. METHODS: A total of 600 (Male, 238 and Female, 362) aging 20-80 Years patients with clinical signs of prolonged cough, chest pain and fever, were evaluated for peripheral blood parameters using hematology analyzer. All the informations related to the disease were collected from the patients and recorded using predesigned questionnaire. RESULTS: Erythrocytic Sedimentation Rate (ESR), Hemoglobin (Hb) and lymphocytes were markedly changed in both sexes. Hemoglobin was recorded lower than normal value in 55% and 53% of male and female population respectively. Total leukocyte count was also lower than normal values in 8% and 6% of male and female respectively. Similarly neutropenia was observed in 5% and 8% cases, while neutrophilia was recorded as 60% and 64% in male and female patients respectively. Lymphocytopenia was also observed in 59% and 43% patients in male and female respectively. Illiteracy, smoking habits, overcrowding and living in shared houses were the main associated risk factors contributing in the enhancement of the disease. CONCLUSION: The disease was present significantly more in females and was relatively higher in older patients. Different hematological parameters like Erythrocytic sedimentation Rate (ESR), platelets and leukocytes work as hallmark and help the clinicians in early diagnosis of the disease. Malnutrition, smoking tobacco, living in shared houses, illiteracy and poverty were the common risk factors contributing to the dissemination of the tuberculosis in the target area population.

Epidemiology, sero-diagnosis and therapeutic studies on nematodes infection in balochi range-sheep at district quetta, balochistan, pakistan.

BACKGROUND: Among the infectious organisms of parasitic origin, gastrointestinal nematodes are very important as they have been reported worldwide. The main aim of the present research study to highlight the annual epidemiological contributing factors associated with the prevalence of gastrointestinal nematodes and their control in sheep. METHODS: A total 1200 faecal samples (100 per month) were collected from farmers holding Balochi-sheep (either sexes, 1-5 years old) during January-December 2012 and analyzed to determine the prevalence of nematodes based on microscopy and ELISA based diagnostic assay. Therapeutic efficacies of different synthetic and herbal medicines against these nematodes were assessed by field trials. RESULTS: Results showed that 23.92% Balochi-sheep were infected with nematodes. Five nematodes infections were recorded with highest prevalence of Haemonchus (7.75%) followed by Nematodirus (7.58%), Strongyloides (4.42%), Trichostrongylus (2.33%) and Trichuris (1.83%). The younger and older ewes (one and five years) presented higher nematodes prevalence with peak during March/April and August/September. Haemonchus and Trichuris positive samples based on coprological examination were also showed 92-100% positive sensitivity for these nematodes by the ELISA. Sheep treated with Ivermectin showed higher reduction (97.76%) in nematode egg counts followed by Atreefal deedan (96.42%) and Oxfendazole (95.44%), respectively. CONCLUSION: The gastro-intestinal nematodes are prevalent in all age and either sex of Balochi-sheep with peak during summer. The ELISA based diagnosis is more accurte. The synthetic and herbal products are very effective against sheep nematodes.

Typhoid fever in paediatric patients in Quetta, Balochistan, Pakistan.

OBJECTIVES: To determine the seropositivity of typhoid fever in febrile pediatric patients presenting to tertiary care center. METHODS: This observational study was conducted at Children Hospital Quetta (CHQ) from July 2011 to March 2012. The children with three or more days fever, no obvious focus of infection and clinically suspected of typhoid fever were screened. Sterile Blood samples were obtained from febrile patients and Widal and Typhidot® tests were performed for the diagnosis of Typhoid fever in the suspected populations. RESULTS: Total of 2964 clinically suspected patients were screened for typhoid fever. Of these, 550 (18.6%) patients were positive serologically. The higher prevalence of the disease in hot summer season and increasing pattern of the disease was observed in summer days. The disease was higher in school age children under 5-10 years. Although non-significant association was observed on sex basis. CONCLUSION: The findings highlight the considerable burden of typhoid fever in pre-school and school-aged children. The variation in the disease pattern has also been observed under seasonal variation and different age groups, all of which need to be considered in deliberations to control the typhoid fever.

Genetic diversity of Newcastle disease virus in Pakistan: a countrywide perspective.

BACKGROUND: Newcastle disease (ND) is one of the most deadly diseases of poultry around the globe. The disease is endemic in Pakistan and recurrent outbreaks are being reported regularly in wild captive, rural and commercial poultry flocks. Though, efforts have been made to characterize the causative agent in some of parts of the country, the genetic nature of strains circulating throughout Pakistan is currently lacking. MATERIAL AND METHODS: To ascertain the genetics of NDV, 452 blood samples were collected from 113 flocks, originating from all the provinces of Pakistan, showing high mortality (30-80%). The samples represented domesticated poultry (broiler, layer and rural) as well as wild captive birds (pigeons, turkeys, pheasants and peacock). Samples were screened with real-time PCR for both matrix and fusion genes (1792 bp), positive samples were subjected to amplification of full fusion gene and subsequent sequencing and phylogenetic analysis. RESULTS: The deduced amino acid sequence of the fusion protein cleavage site indicated the presence of motif (112RK/RQRR↓F117) typical for velogenic strains of NDV. Phylogenetic analysis of hypervariable region of the fusion gene indicated that all the isolates belong to lineage 5 of NDV except isolates collected from Khyber Pakhtunkhwa (KPK) province. A higher resolution of the phylogenetic analysis of lineage 5 showed the distribution of Pakistani NDV strains to 5b. However, the isolates from KPK belonged to lineage 4c; the first report of such lineage from this province. CONCLUSIONS: Taken together, data indicated the prevalence of multiple lineages of NDV in different poultry population including wild captive birds. Such understanding is crucial to underpin the nature of circulating strains of NDV, their potential for interspecies transmission and disease diagnosis and control strategies.

Prevalence of bovine brucellosis in organized dairy farms, using milk ELISA, in quetta city, balochistan, pakistan.

A total of 200 milk samples from cattle (n = 86) and buffalo (n = 114) were evaluated using milk ring test (MRT) and indirect enzyme-linked immunosorbent assay (i-ELISA). The overall prevalence was found to be 3% and 8.5% in cattle and buffaloes using MRT and i-ELISA, respectively. The prevalence was 4.6% and 1.7% in cattle and buffalo using MRT, respectively, while i-ELISA exhibited 20% and 0% in cattle and buffalo, respectively. The prevalence was higher in government dairy farm, compared to privately owned dairy farm. This paper points out an alarming situation in the target area with respect to the public health significance.