Genetic Characterization of Carbapenem Resistant Acinetobacter baumannii in Tertiary care settings of Lahore, Pakistan

Background: Acinetobacter baumannii is major cause of ventilator associated pneumoniae (VAP) as it is an opportunistic nosocomial organism. The current study was to find out the antibiotic resistance pattern of Acinetobacter baumannii, its phenotype and the genetic characterization of Metallo-β-Lactamase (MBL) genes that are responsible for carbapenem resistance.Methods: One hundred and fifty Carbapenem resistant Acinetobacter baumannii (CRAB) specimens were isolated and PCR amplification of organism specific bla-OXA-51gene was performed and antibiotic susceptibility was checked. Phenotypic susceptibility analysis was performed by Modified Hodge Test (MHT) and Imipenem-EDTA Double Disc Synergy Test (IMP-EDTA DDST). The carbapenemases and MBL producing genes were amplified by PCR.Results: CRAB showed high resistance against piperacillin/tazobactam (99.3%), cefepime and ceftazidime (99.3% each), amikacin (91.3%), ciprofloxacin (96.7%) and levofloxacin (96.7%). Only one isolate showed resistance to colistin. The isolates positive for both MHT and DDST (n=70) were further characterized to detect metallo-β-lactamase genes. Molecular characterization revealed the presence of bla-OXA-51 gene in all tested isolates (100%) followed by bla-VIM 89%, bla-OXA-23 64%, respectively and so on. Few genes coexisted with each other including bla VIM, bla OXA 23, bla OXA 51 and bla NDM-1.  None of the isolate was found positive for bla-IMP gene.Conclusion: It is concluded that CRAB isolates exhibited a high rate of resistance towards antimicrobials because of the presence of drug hydrolyzing enzymes, carbapenemases and MBLs. This is among the rare study reported recently indicating CRAB isolates co-harboring many resistant genes are very difficult to treat. There is a dire need to develop novel antibiotics against resistant A. baumannii to minimize its prevalence. Moreover, it is recommended that colistin treatment in the clinical settings should be continuously monitored in order to prevent the development of resistance

Enhanced production of butyric acid by solid-state fermentation of rice polishings by a mutant strain of Clostridium tyrobutyricum

Purpose: To enhance butyric acid production by solid-state fermentation with a hyper-producing mutant of Clostridium tyrobutyricum generated by random mutagenesis.Methods: Wild type C. tyrobutyricum was mutagenized with UV irradiation, nitrous acid, and ethidium bromide to obtain a hyper-producing strain. Various physiochemical parameters were optimized to increase the butyric acid yield.Results: The UV-induced mutant (C.TUV) produced significantly higher concentrations of butyric acid than the wild type parent, nitrous acid-induced, and ethidium bromide-induced strains. C.TUV increased butyric acid production 1.4-fold more than the parent strain. Fermentation with C.TUV with 2.5 g of rice polishings (w/w), a 2 % inoculum volume (v/v), and a 48-h incubation period at 37°C under anaerobic conditions produced 11.63 mg/100 g of butyric acid. The addition of 0.6 % corn steep liquor as a nitrogen source increased the butyric acid concentration to 26.09 mg/100 g.Conclusion: These optimized fermentation parameters on a small scale can be used on a commercial scale to mass-produce butyric acid.Keywords: Butyric acid, Mutant, Clostridium tyrobutyricum, Mutagen, Solid-state fermentatio

Reconnoitering Milk Constituents of Different Species, Probing and Soliciting Factors to Its Soundness

Milk composition and production varies from species to species, reflecting its diversified benefits on health. Lipids from caprine and ovine milk are anti-obesity and anti-atherogenic while prebiotic in the case of caprine. Higher contents of selenium from caprine and iron from camel milk play a role in immune system and oxygen transport system, respectively, whereas enriched vitamins like riboflavin, folic acid, B6, vitamin A of bovine, and foliate of cattle are effective in the synthesis of hemoglobin, and high niacin content of caprine is anti-cancerous. Camel milk is found to have characteristics of anti-carcinogenic, antidiabetic, and autoimmune therapeutic. Various processing techniques like pasteurization, skim milk powder processing, and ultra-high temperature processing are necessary for safe provision of milk to meet consumers’ demand. Change in flavor, loss of micronutrients, biofilm production, and spore-forming bacteria are prominent challenges during processing. Antimicrobial resistance and disease conditions are exaggerating factors of milk deterioration with respect to quality and quantity. Preclinical trials like somatic cell count, California mastitis test, proteomic analysis, Raman spectroscopy-based analysis, and X-ray fluorescence analysis are helpful in avoiding the spread of disease and controlling of economic losses. This chapter focuses differential functions of bioactive of milk, issues arising during processing techniques, and preclinical studies of milk for safer production and consumption of milk

Microbial profiling and risk factors assessment for Otitis Media and Otitis Externa

Background: Otitis media and otitis externa are common otological manifestations in all generations especially in children. There is lack of accurate identification of the causative agent and thus poor diagnosis for such infections. Therefore, it leads to permanent anatomical disabilities including poor speech and defects in balancing and hearing. The study was conducted to isolate, characterize and identify the microbes causing otitis media and otitis externa. Methods: A total of 250 patients having otitis media and otitis externa were enrolled in the study from March 2011 to October 2011. All patients were examined through clinical examination and detailed history was collected. Pus samples from the discharging ears were plated on MacConkey’s, Chocolate and Blood agar for 24 to 48 hours. Isolates were identified on the basis of morphology, staining reactions and various biochemical tests. Results: In this study, only 6% cases yielded no growth, 14% yielded mixed cultures while 80% cases yielded pure cultures. The presumptive diagnosis for ear swabbing was otitis media (76%) and otitis externa (24%). The most common bacterial isolates obtained were Staphylococcus aureus (43.3%) followed byPseudomonas aeruginosa (25%) in the diagnosed cases of otitis media. While for the cases of otitis externa,Pseudomonas aeruginosa was the predominant organism with 52.2%. Infection of otitis media was most common among children and the persons having low socioeconomic conditions. Conclusion: Pseudomonas aeruginosa was identified as the principal pathogen followed by Staphylococcus aureus. To circumvent the painful effects of acute and chronic ear infections, an accurate microbial profiling may play pivotal role

Factors Affecting the Service Quality and Customer Satisfaction in Telecom Industry of Pakistan

The current study focuses on the service quality factors which influence customer satisfaction in telecom industry of Pakistan. Customer satisfaction can be built up when brand satisfies the requirements and cravings of clients. Therefore, we took five service quality dimensions (empathy, reliability, assurance, responsiveness & tangibility) as predicting variables towards criterion variable customer satisfaction. This research was carried out on customers of telecom sector. An aggregate of 320 questionnaires were distributed among the customers of telecom industry of Pakistan through electronic forms as well as simple forms out of which 255 forms were returned i.e. response rate was79.7%. The outcomes of statistical tests demonstrate that three out of five predictor variables i.e. assurance, responsiveness and reliability have positive impact on customer satisfaction while empathy and tangibility have no significant influence on customer satisfaction. Research limitations and future plans also discussed in this research

Etiological and Clinical Spectrum of Acute Liver Failure of Infancy in Pakistan

  Objective: To describe the aetiology and clinical spectrum of acute liver failure of infancy at a tertiary care hospital Study Design: Cross-sectional study. Place and Duration of Study: Department of Paediatric Gastroenterology, Hepatology & Nutrition, Children Hospital and Institute of Child Health, Lahore, from Nov 2020 to May 2021. Methodology: Infants under 12 months of age were enrolled having liver-based coagulopathy (not corrected after two doses of parenteral vitamin K, 10 mg) with INR > 2, whether encephalopathy was present or not. Encephalo-pathy is difficult to identify in infants, so it was not essential for the diagnosis of ALFI in our study. Infants diagnosed with chronic liver disease at presentation or those without final etiological diagnosis were excluded. Results: A total of 31 infants were enrolled fulfilling the criteria of acute liver failure of infancy and were studied about aetiology and clinical presentation. The mean age of presentation was 4.64±3.16 months, and males predominated in the study group (64.5%). Common clinical features were in descending order ascites in 29 (93.5%), jaundice in 28 (90.3%), pallor in 24 (77.4%) and peripheral oedema in 21 (67.7%). Metabolic liver diseases were the common cause of ALFI, constituting around(18, 58%) followed by sepsis (9, 29%).Galactosemia (11, 35.5%) stands out among the metabolic causes. Conclusion: Metabolic disorders followed by sepsis are the most common cause of ALFI

Augmented cytotoxic, mutagenic and genotoxic response triggered by carvedilol and celecoxib combinations

It is understood that drugs regardless of their order of administration can exhibit drug interactions. Established on the fact that treatment of hypertension may last for decades and prolong usage of multiple drug regimen may induce substantial pathophysiological changes. Hence, This study was designed to evaluate the possible synergistic toxic effects of anti-hypertensive (carvedilol), and anti-inflammatory drug (celecoxib) alone and in combinations. Well-established MTT assay, Single Cell Gel Electrophoresis (SCGE) and Ames assay were employed to evaluate the toxicity at cellular level. Results from MTT assay on Vero cell line revealed that drug combinations have more pronounced anti-proliferative activity with combine IC50 value of 13.7:47.8 µg/mL. Likewise, exposure of peripheral blood mononuclear cells with drug combinations revealed significant (

Prevalence of Black Quarter in Association with Patho-Morphological Alterations in Affected Tissues at Different Zones of Punjab-Pakistan

Background: Blackleg or Black Quarter (BQ) is a serious bacterial disease caused by Clostridium chauvoei. It causes edematous and gaseous changes in skeletal muscles of animals. The study was designed to find prevalence of BQ in 6 districts of Punjab, Pakistan from June 2018 to June 2019.Methods: Animals were randomly monitored, and selected in each union council regardless of age, sex and species, but lameness and hyperthermia were the selection parameters. A proforma was used to record the experimental readings of each animal. A total of 1500 swab samples from bovines (cattle and buffaloes) were processed through conventional PCR and culture sensitivity tests to examine the comparative diagnostic efficacy and suitability of the test.Results: Sensitivity and specificity of culture tests were 87.6% and 34.19% respectively, whereas PCR demonstrated 81% and 47.3% results for the said aspects, respectively. Alpha toxin gene (Ccta) was amplified at 52.2oC yielding an amplicon size of 1400 base pairs. Area wise and season wise prevalence of each animal was also determined. The prevalence of BQ in cattle was significantly higher (P 0.05) in Lahore (0%) and Nankana Sahib (8%) which belonged to the canal irrigated zones.Conclusion: There was higher prevalence of BQ in different areas of Punjab, Pakistan that show the higher risk of disease transmission. A systematic surveillance system is essential to regularly monitor the disease incidence and prevalence in these areas of Pakistan.Keywords: Black Quarter; Prevalence; Pathological; Ruminants;  Zone