Contribution of the gyrA and waaG mutants to fluoroquinolones resistance, biofilm development, and persister cells formation in Salmonella enterica serovar Typhi.

Fluoroquinolone resistance in Salmonella has been reported worldwide and poses a serious public health threat in developing countries. Multiple factors contribute to fluoroquinolone resistance, including mutations in DNA gyrase and the acquisition of antimicrobial resistance genes. Salmonella enterica serovar Typhi (S. Typhi) causes typhoid fever in humans, which is highly prevalent in counties with poor sanitation and hygiene standards. Here, we reported S. Typhi clinical isolates that showed varying degrees of susceptibility to fluoroquinolones and were characterized by Analytical Profile Index 20E test kit and 16S rRNA sequencing. S. Typhi strain S27 was resistant to fluoroquinolones and had multiple mutations in the gyrA gene. The gyrA lies in the quinolone resistance determining region of S. Typhi and has mutations at codon 83 (Ser83Phe), codon 87 (Asp87Gly), codon 308 (Lys308Glu), and codon 328 (Val328Ile). S. Typhi strain S6 has no gyrA mutations and is sensitive to fluoroquinolones but forms a strong biofilm relative to S. Typhi S27. Transcriptional analysis of biofilm associated genes revealed that the waaG gene was significantly downregulated. The ΔwaaG mutant showed a significant decrease in persister cells and a strong biofilm formation relative to wild type and gyrA mutant. The gyrA tetra mutant persister assay revealed a significant increase in persister cells compared to wild type and ΔwaaG. Collectively, this is the first report of S. Typhi's two key genes and their roles in antibiotic tolerance, biofilm formation, and fluoroquinolone resistance that can help in understanding the mechanism of persister formation and eradication.

Teicoplanin associated gene tcaA inactivation increases persister cell formation in Staphylococcus aureus.

Staphylococcus aureus is part of normal human flora and is widely associated with hospital-acquired bacteremia. S. aureus has shown a diverse array of resistance to environmental stresses and antibiotics. Methicillin-resistant S. aureus (MRSA) is on the high priority list of new antibiotics discovery and glycopeptides are considered the last drug of choice against MRSA. S. aureus has developed resistance against glycopeptides and the emergence of vancomycin-intermediate-resistant, vancomycin-resistant, and teicoplanin-resistant strains is globally reported. Teicoplanin-associated genes tcaR-tcaA-tcaB (tcaRAB) is known as the S. aureus glycopeptide resistance operon that is associated with glycopeptide resistance. Here, for the first time, the role of tcaRAB in S. aureus persister cells formation, and ΔtcaA dependent persisters' ability to resuscitate the bacterial population was explored. We recovered a clinical strain of MRSA from a COVID-19 patient which showed a high level of resistance to teicoplanin, vancomycin, and methicillin. Whole genome RNA sequencing revealed that the tcaRAB operon expression was altered followed by high expression of glyS and sgtB. The RNA-seq data revealed a significant decrease in tcaA (p = 0.008) and tcaB (p = 0.04) expression while tcaR was not significantly altered. We knocked down tcaA, tcaB, and tcaR using CRISPR-dCas9 and the results showed that when tcaA was suppressed by dCas9, a significant increase was witnessed in persister cells while tcaB suppression did not induce persistence. The results were further evaluated by creating a tcaA mutant that showed ΔtcaA formed a significant increase in persisters in comparison to the wild type. Based on our findings, we concluded that tcaA is the gene that increases persister cells and glycopeptide resistance and could be a potential therapeutic target in S. aureus.

The functional repertoire of AmpR in the AmpC ß-lactamase high expression and decreasing ß-lactam and aminoglycosides resistance in ESBL Citrobacter freundii.

Citrobacter freundii is characterized by AmpC ß-lactamases that develop resistance to ß-lactam antibiotics. The production of extended-spectrum ß-lactamase (ESBL) is substantially high in Escherichia coli, C. freundii, Enterobacter cloacae, and Serratia marcescens, but infrequently explored in C. freundii. The present investigation characterized the ESBL C. freundii and delineated the genes involved in decrease in antibiotics resistance. We used the VITEK-2 system and Analytical Profile Index (API) kit to characterize and identify the Citrobacter isolates. The mRNA level of AmpC and AmpR was determined by RT-qPCR, and gel-shift assay was performed to evaluate protein-DNA binding. Here, a total of 26 Citrobacter strains were isolated from COVID-19 patients that showed varying degrees of antibiotic resistance. We examined and characterized the multidrug resistant C. freundii that showed ESBL production. The RT-qPCR analysis revealed that the AmpC mRNA expression is significantly high followed by a high level of AmpR. We sequenced the AmpC and AmpR genes that revealed the AmpR has four novel mutations in comparison to the reference genome namely; Thr64Ile, Arg86Ser, Asp135Val, and Ile183Leu while AmpC remained intact. The ΔAmpR mutant analysis revealed that the AmpR positively regulates oxidative stress response and decreases ß-lactam and aminoglycosides resistance. The AmpC and AmpR high expression was associated with resistance to tazobactam, ampicillin, gentamicin, nitrofurantoin, and cephalosporins whereas AmpR deletion reduced ß-lactam and aminoglycosides resistance. We conclude that AmpR is a positive regulator of AmpC that stimulates ß-lactamases which inactivate multiple antibiotics.

Eco-friendly textile desizing with indigenously produced amylase from Bacillus cereus AS2.

Starch is added to the fabric surface to secure weaving process. During finishing these sized particles are removed from the fabric and prepared it for printing and dyeing. Chemicals de-sizing agents damage fabric surfaces and reduce the quality of the product. An alternative to these conventional desizing agents is the use of biological molecules i.e. enzymes. The current study compares traditional de-sizing to bio-based de-sizing methods, as well as the optimization of fabric desizing settings using crude amylase. Amylase-producing Bacillus cereus AS2 was isolated from indigenous soil samples. The maximal fermentative de-sizing capability was discovered at 72 h, with no fabric surface degradation. Chemical desizing showed that the fabric lost all sizing agents to TEGEWA scale 9 within 1 h in presence of 5N HCl. Optimal studies for desizing showed that 1000 IU/ml of amylase resulted in maximum de-sizing within 15 h at 60 °C and 0.5% Triton-X. Water absorbance and weight loss, both parameters were used to check the desizing efficacy and it was found that de-sizing to same scale was occurred in the case of enzyme as well as commercially desized fabric. Enzyme desized cloth was found to be free of any starch particles in SEM micrographs, identical to industrially de-sized fabric, ensuring bioprocess efficacy.

Clinical manifestations of active tuberculosis patients coinfected with severe acute respiratory syndrome coronavirus-2.

Summary background: The coronavirus 2019 pandemic was caused by a new single-strand RNA virus that originated from Wuhan, China, and infected more than 190 countries. The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) coinfection with tuberculosis posed a serious public health concern and complicated the prognosis and treatment of patients. Since both are respiratory diseases, the sign and symptoms may overlap and could have synergistic effects on the host that can increase mortality during coinfection. The present investigation reported the clinical characteristics of patients having coinfection of COVID-19 and tuberculosis (COVID-TB). Methods: We performed a retrospective investigation on COVID-19 infection in tuberculosis patients between the years 2020 and 2021. The SARS-CoV-2 was confirmed by PCR and the COVID-TB epidemiological and clinical findings were recorded on the day of admission and followed up for 25 days. Results: The mean age of the COVID-19 patients was 50 ± 15 years, 76.36% were male and 23.64% were female. Weight loss, sore throat, whooping cough, chest pain, and vomiting were common symptoms, and asthma, diabetes, arthritis, and hypertension were found as co-morbidities in COVID-TB. The D-dimer, lactate dehydrogenase, C-reactive protein, erythrocyte sedimentation rate, and creatine kinase levels increased 14-fold, 12.5-fold, 11-fold, 10-fold, and 7-fold respectively during COVID-TB. The patients suffered from hyperferritinemia and lymphocytopenia which increased the likelihood of death. The levels of D-dimer, lactate dehydrogenase, C-reactive protein, erythrocyte sedimentation rate, and creatinine kinase were positively correlated with patient age. The chest radiograph showed the infectious agents have consolidated opacity and peripheral dissemination in the lungs. Conclusion: Tuberculosis coinfection augmented the severity of COVID-19 and the likelihood of death, and high vigilance is recommended for respiratory pathogens in COVID-19.

Study on awareness, knowledge, and practices towards antibiotic use among the educated and uneducated people of Khyber Pakhtunkhwa Province, Pakistan

INTRODUCTION: Antimicrobial resistance developed through the inadequate use of antibiotics; is an overriding task for global public health. OBJECTIVE: To explore awareness, knowledge, and practices, and compare the elements associated with antibiotic misuse in different University students and uneducated people of Khyber Pakhtunkhwa Province, Pakistan. METHODS: Cross-sectional study was conducted from July to December 2020 using a validated questionnaire. Data were collected from eleven different university students and uneducated people of Khyber Pakhtunkhwa, Pakistan. RESULTS: 3,600 questionnaires were completed, consisting of 56.9% Male and 43.0% Female. 1,999 (55.5%) of the antibiotic users reported through the survey used non-prescription antibiotics within a one-month study period. Out of the participants, 230 (6.3%) were uneducated or their education level was below matric rest were university students. 1999 (55.5%) reported buying Antibiotics with Medical Prescription. Most self-medicated participants (56.9%) stop taking antibiotics when they feel better. More than 90% of the respondents answered that doctors and pharmacist staff do not guide them well that how to use antibiotics. 2,171 (60.03%) respondents mistakenly believed that antibiotics improve restoration from coughs and colds. Only 720 (20%) respondents knew that antibiotics also disturb normal flora and 547 participants (15.9%) agree that unnecessary use of antibiotics causes bacterial resistance. CONCLUSION: Finding from this study may have important implications for public health policy in Khyber Pakhtunkhwa, Pakistan given the growing global resistance to antibiotics and the reported health issues related to their improper use.

Myogenesis and Analysis of Antimicrobial Potential of Silver Nanoparticles (AgNPs) against Pathogenic Bacteria.

The widespread and indiscriminate use of broad-spectrum antibiotics leads to microbial resistance, which causes major problems in the treatment of infectious diseases. However, advances in nanotechnology have opened up new domains for the synthesis and use of nanoparticles against multidrug-resistant pathogens. The traditional approaches for nanoparticle synthesis are not only expensive, laborious, and hazardous but also have various limitations. Therefore, new biological approaches are being designed to synthesize economical and environmentally friendly nanoparticles with enhanced antimicrobial activity. The current study focuses on the isolation, identification, and screening of metallotolerant fungal strains for the production of silver nanoparticles, using antimicrobial activity analysis and the characterization of biologically synthesized silver nanoparticles by X-ray diffraction (XRD) spectroscopy, energy-dispersive X-ray spectroscopy (EDX), and scanning electron microscopy (SEM). In total, 11 fungal isolates were isolated and screened for the synthesis of AgNPs, while the Penicillium notatum (K1) strain was found to be the most potent, demonstrating biosynthetic ability. The biologically synthesized silver nanoparticles showed excellent antibacterial activity against the bacteria Escherichia coli (ATCC10536), Bacillus subtilis, Staphylococcus aureus (ATCC9144), Pseudomonas aeruginosa (ATCC10145), Enterococcus faecalis, and Listeria innocua (ATCC13932). Furthermore, three major diffraction peaks in the XRD characterization, located at the 2θ values of 28.4, 34.8, 38.2, 44, 64, and 77°, confirmed the presence of AgNPs, while elemental composition analysis via EDX and spherical surface topology with a scanning electron microscope indicated that its pure crystalline nature was entirely composed of silver. Thus, the current study indicates the enhanced antibacterial capability of mycologically synthesized AgNPs, which could be used to counter multidrug-resistant pathogens.

Bioactive, antioxidant and antimicrobial properties of chemically fingerprinted essential oils extracted from Eucalyptus globulus: in-vitro and in-silico investigations.

Innovative approaches are urgently required to treat divestating bacterial diseases in the face of rising bacterial resistance rates. The current investigative work focused on hydro-distilling Tasmanian blue gum (Eucalyptus globulus) to isolate the essential oil, which was then tested for bioactivity, antioxidant capacity, and antibacterial activity using in-vitro and in silico assays. The antioxidant activity was avualated against DPPH and FRAP. With results of 69.63 RSA (%) (µL/L AAE) at a concentration of 5 mL/L and 51.56 (µL/L AAE) at concentration of 90 ppm in the 2,20-diphenyl-1-picryl hydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, respectively, the extracted oil indicated considerable antioxidant activity. The extracted oil demonstrated powerful antibacterial activity in in-vitro tests against both Gram-positive and Gram-negative bacterial strains, including Bordetella bronchiseptica (21 mm), Staphylococcus epidermidis (19 mm), and Staphylococcus aureus (19 mm), with significant minimal inhibitory (MIC) and minimum bactericidal (MIB) concentrations. Additionally, GC-MS analysis of the oil from E. globulus identified several low-molecular-weight compounds, including Eucalyptol, γ-Terpinene, Shisool acetate, 1,3-trans,5-cis-Octatriene, 2,6-Dimethyl-1,3,5,7-octatetraene, E,E, Cyclohexene, 1-methyl-4-(1-methylethylidene), Benzene, 1-methyl-4-(1-methylethenyl), Butanoic acid, 3-methyl-, 3-methylbutyl ester, and 1,3,8-p-Menthatriene. Several other compounds were also identified, including Fenchol, 2-Methyl-trans-3a,4,7,7a-tetrahydroindane, (E,E,E)-2,4,6-Octatriene, 1,2,3,6-Tetrahydrobenzylalcohol, acetate, Alloaromadendrene, Phenol, 2-ethyl-4,5-dimethyl, Phenol, 2-methyl-5-(1-methylethyl)-, p-Cymen-7-ol, 1,5,5-Trimethyl-6-methylene-cyclohexene, 1,3-Cyclohexadiene, 1-methyl-4-(1-methylethyl)-, 2,6-Octadien-1-ol, 3,7-dimethyl-, acetate, (Z), and more. The bioactive potential of Eucalyptus globulus essential oil against 1AJ6 and 1R4U was highlighted by molecular docking analyses, suggesting its utility as a natural source of antioxidant and antibacterial compounds with the potential to replace chemical disinfectants in a variety of applications.

High-Molecular-Weight Dextran-Type Exopolysaccharide Produced by the Novel Apilactobacillus waqarii Improves Metabolic Syndrome: In Vitro and In Vivo Analyses.

Metabolic syndrome is a leading medical concern that affects one billion people worldwide. Metabolic syndrome is defined by a clustering of risk factors that predispose an individual to cardiovascular disease, diabetes and stroke. In recent years, the apparent role of the gut microbiota in metabolic syndrome has drawn attention to microbiome-engineered therapeutics. Specifically, lactic acid bacteria (LAB) harbors beneficial metabolic characteristics, including the production of exopolysaccharides and other microbial byproducts. We recently isolated a novel fructophilic lactic acid bacterium (FLAB), Apilactobacillus waqarii strain HBW1, from honeybee gut and found it produces a dextran-type exopolysaccharide (EPS). The objective of this study was to explore the therapeutic potential of the new dextran in relation to metabolic syndrome. Findings revealed the dextran's ability to improve the viability of damaged HT-29 intestinal epithelial cells and exhibit antioxidant properties. In vivo analyses demonstrated reductions in body weight gain and serum cholesterol levels in mice supplemented with the dextran, compared to control (5% and 17.2%, respectively). Additionally, blood glucose levels decreased by 16.26% following dextran supplementation, while increasing by 15.2% in non-treated mice. Overall, this study displays biotherapeutic potential of a novel EPS to improve metabolic syndrome and its individual components, warranting further investigation.

Response Surface Methodology for Optimization of Operational Parameters To Remove Ciprofloxacin from Contaminated Water in the Presence of a Bacterial Consortium.

Ciprofloxacin (CFX) is a broad-spectrum fluoroquinolone antibiotic that is widely used to treat bacterial infections in humans and other animals. However, its unwanted occurrence in any (eco)system can affect nontarget bacterial communities, which may also impair the performance of the natural or artificially established bioremediation system. The problem could be minimized by optimization of operational parameters via modeling of multifactorial tests. To this end, we used a Box-Behnken design in response surface methodology (RSM) to generate the experimental layout for testing the effect of the CFX biodegradation for four important parameters, that is, temperature (°C), pH, inoculum size (v/v %), and CFX concentration (mg L-1). For inoculation, a consortium of three bacterial strains, namely, Acenitobacter lwofii ACRH76, Bacillus pumilus C2A1, and Mesorihizobium sp. HN3 was used to degrade 26 mg L-1 of CFX. We found maximum degradation of CFX (98.97%; initial concentration of 25 mg L-1) at 2% inoculum size, 7 pH, and 35 °C of temperature in 16 days. However, minimum degradation of CFX (48%; initial concentration of 50 mg L-1) was found at pH 6, temperature 30 °C, and inoculum size 1%. Among different tested parameters, pH appears to be the main limiting factor for CFX degradation. Independent factors attributed 89.37% of variation toward CFX degradation as revealed by the value of the determination coefficient, that is, R 2 = 0.8937. These results were used to formulate a mathematical model in which the computational data strongly correlated with the experimental results. This study showcases the importance of parameter optimization via RSM for any bioremediation studies particularly for antibiotics in an economical, harmless, and eco-friendly manner.

Pathophysiology of Methicillin-Resistant Staphylococcus aureus Superinfection in COVID-19 Patients.

The global spread of the coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has infected humans in all age groups, deteriorated host immune responses, and caused millions of deaths. The reasons for individuals succumbing to COVID-19 were not only the SARS-CoV-2 infection but also associated bacterial infections. Antibiotics were largely used to prevent bacterial infections during COVID-19 illness, and many bacteria became resistant to conventional antibiotics. Although COVID-19 was considered the main culprit behind the millions of deaths, bacterial coinfections and superinfections were the major factors that increased the mortality rate in hospitalized patients. In the present study, we assessed the pathophysiology of methicillin-resistant Staphylococcus aureus (MRSA) superinfection in COVID-19 patients in Pakistan. A total of 3492 COVID-19 hospitalized patients were screened among which 224 strain were resistant to methicillin; 110 strains were tazobactam-resistant; 53 strains were ciprofloxacin-resistant; 23 strains were gentamicin-resistant; 11 strains were azithromycin-resistant; 3 strains were vancomycin-resistant. A high frequency of MRSA was detected in patients aged ≥50 with a prevalence of 7.33%, followed by patients aged >65 with a prevalence of 5.48% and a 5.10% prevalence in patients aged <50. In addition, pneumonia was detected in the COVID-19-associated MRSA (COVID-MRSA) that showed decreased levels of lymphocytes and albumin and increased the mortality rate from 2.3% to 25.23%. Collectively, an MRSA superinfection was associated with increased mortality in COVID-19 after 12 to 18 days of hospitalization. The study assessed the prevalence of MRSA, mortality rate, pneumonia, and the emergence of antibiotic resistance as the main outcomes. The study summarized that COVID-MRSA aggravated the treatment and recovery of patients and suggested testing MRSA as critical for hospitalized patients.

Effect of ellagic acid and mesocarp extract of Punica granatum on productive and reproductive performances of laying hens.

In the present study, we determined the potential effects of ellagic acid and mesocarp extract of Punica granatum on the productive and reproduction performance of laying hens. Five treatment groups were setup: (1) control group (without ellagic acid), (2) 50 mg of ellagic acid, (3) 100 mg of ellagic acid, (4) 200 mg of ellagic acid, and (5) mesocarp extract of P. granatum. All the groups were investigated for feed intake, body weight, egg production, egg quality, fertility, hatchability, antioxidant status of serum and liver, lipid peroxidation, and antibacterial activities. Egg production, feed intake, and bodyweight were significantly increased (p < 0.05) with 100 mg of ellagic acid and P. granatum extract while no significant effect was observed on albumen and yolk weight, yolk index, yolk color, egg-shape index, and Haugh unit. Both ellagic acid and P. granatum extract significantly improved hatchability while 100 and 200 mg/kg of ellagic acid numerically decreased fertility. Besides, ellagic acid (100 mg/kg) and P. granatum extract significantly decreased malondialdehyde concentration and increased total antioxidant capacity, glutathione peroxidase, and total superoxide dismutase in serum and liver samples of laying hens (p < 0.05). The lipid peroxidation was decreased among the treatment groups, with 100 mg of ellagic acid and P. granatum extract showed the best activity. Moreover, ellagic acid demonstrated strong killing activity against Escherichia coli and Staphylococcus aureus while it was ineffective against methicillin-resistant S. aureus. Our results conclude that ellagic acid and P. granatum promoted egg production, hatchability, and antioxidant enzyme activities of the laying hens.

Production of bimodal molecular weight levan by a Lactobacillus reuteri isolate from fish gut.

An exopolysaccharide (EPS) synthesizing potentially probiotic Gram-positive bacterial strain was isolated from fish (Tor putitora) gut, and its EPS was structurally characterized. The isolate, designated as FW2, was identified as Lactobacillus reuteri through 16S rRNA gene sequencing and phylogenetic analysis. This isolate produces fructan-type EPS using sucrose as a substrate. Based on 13C-NMR spectroscopy, methylation analysis and monosaccharide composition, the EPS was identified as a linear levan polymer with fructose as main constituent linked via ß(2 → 6) linkages. Based on molecular weight (MW) distribution, two groups of levan were found to be produced by the isolate FW2: one with high MW (4.6 × 106 Da) and the other having much lower MW (1.2 × 104 Da). The isolate yielded about 14 g/L levan under optimized culturing parameters including aeration conditions, pH, temperature and substrate concentration. The obtained bimodal molecular weight linear levan is the first of its type to be synthesized by a L. reuteri isolate from fish gut. Bimodal molecular weight prebiotic levan together with the probiotic potential of the producing strain would provide a new promising synbiotic combination for use in aqua culture.

Growth Inhibition of Common Enteric Pathogens in the Intestine of Broilers by Microbially Produced Dextran and Levan Exopolysaccharides.

Antibiotics are generally applied for treatment or as subtherapeutic agents to overcome diseases caused by pathogenic bacteria including Escherichia coli, Salmonella and Enterococcus species in poultry. However, due to their possible adverse effects on animal health and to maintain food safety, probiotics, prebiotics, and synbiotics have been proposed as alternatives to antibiotic growth promoters (AGPs) in poultry production. In this study, the effects of prebiotics on the augmentation of broiler's indigenous gut microbiology were studied. Day old 180 broilers chicks were divided into four treatment groups: G, L, C1, and C2. The groups G and L were fed with basal diet containing 3% dextran and 3% levan, respectively. Control groups were fed with basal diets without antibiotic (C1) and with antibiotics (C2). The experimental groups showed decreased mortality as compared to control groups. After 35 days, the chickens were euthanized and intestinal fluid was analyzed for enteric pathogens on chromogenic agar plates and by 16S rRNA gene sequencing. Inhibition of the growth of E. coli and Enterococcus was observed in groups G and L, respectively, whereas Salmonella was only present in group C1. Also, high populations of lactic acid bacteria were detected in the intestine of prebiotic fed birds as compared to controls. These results depict that dextran and levan have the potential to replace the use of antibiotics in poultry feed for inhibiting the growth of common enteric pathogens. To the best of our knowledge, this is the first study where effects of dextran and levan on intestinal microbiota of broilers have been reported.

Burn Wound Microbiology and the Antibiotic Susceptibility Patterns of Bacterial Isolates in Three Burn Units of Abbottabad, Pakistan.

Infection is the leading cause of morbidity and mortality among burn patients and is accentuated multifold by the emergence of antimicrobial resistance among the nosocomial isolates. It is vital to know the common organisms involved in infected burn wound etiology and their respective antibiotic susceptibility patterns. These crucial findings can help in formulating a better and more efficient antimicrobial therapy plan for controlling burn wound infections. The current study was conducted to identify the common bacteria involved in causing infections in wounds of burn patients and their respective antibiotic susceptibility patterns in three hospitals of Abbottabad, Pakistan. A total of 100 patients were included from the burn units of three hospitals in Abbottabad. Wound swabs were taken from the deepest portions of infected burns, and the organisms involved were isolated via standard microbiological techniques. The Kirby-Bauer disc diffusion technique was used to monitor antibiotic susceptibility. Gram-positive organisms were found readily in infected burn wounds. Staphylococcus aureus (46%) was the most common isolate followed by Staphylococcus epidermidis (17%), Escherichia coli (16%), Proteus spp. (12%), Klebsiella pneumoniae (10%), and Pseudomonas aeruginosa, which was only 7%. Gram-positive bacteria were sensitive to amikacin, gentamicin, cefotaxime, and norfloxacin. In contrast, the gram-negative isolates were sensitive to amikacin, chloramphenicol, and nalidixic acid. Pseudomonas was resistant to most of the antibiotics tested in the present study.

Fasciola hepatica infection in children actively detected in a survey in rural areas of Mardan district, Khyber Pakhtunkhawa province, northern Pakistan.

Human fascioliasis is a freshwater snail borne, zoonotic disease caused by Fasciola liver flukes which are widely spread throughout Pakistan and has recently proved to be endemic in humans of the Punjab province. To verify whether fasciolids are also affecting humans outside this province, studies were conducted in four communities comprising rural and urban areas of Mardan district, Khyber Pakhtunkha province, northern Pakistan. Activities comprised two surveys, a coprological one to look for Fasciola infection and another to get information on potential human infection sources and risk factors by means of a questionnaire. Out of 540 subjects of all ages surveyed, only 4 children (0.74%) were found positive for Fasciola eggs: two 9- and 13-year old boys and two 7- and 16-year old girls. Burden per child ranged 73-146 eggs per gram of faeces (epg), with an average of 106 ±â€¯15.4 epg. Based on the egg size, the infection was diagnosed to be caused by F. hepatica. Serum analyses showed eosinophilia and elevated enzyme values in liver function tests. The Mardan area fits well to all transmission and epidemiological requirements of F. hepatica, namely the presence of infected livestock reservoirs throughout, the appropriate climatic factors (mainly monthly temperature variation along the year), and the existence of adequate freshwater collections. The existence of the specific lymnaeid snail vector species for F. hepatica transmission was reported in the neighbouring Swat valley time ago. Results show that human fascioliasis may be widespread in Pakistan and that both F. hepatica and F. gigantica may be involved.

Prevalence and risk factors of hepatitis B and C viruses among haemodialysis patients: a multicentric study.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are important causes of morbidity and mortality among haemodialysis (HD) patients and create problems in the management of patients in the renal dialysis units, as patients with chronic renal failure do not absolutely clear these viral infections. AIM: The aim of the study is molecular detection of HBV and HCV and their possible risk factors among the HD patients in northern Pakistan. MATERIALS AND METHODS: A cross-sectional study was conducted from November 2013 to June 2014. The infections were investigated through serological and molecular techniques. RESULTS: The overall prevalence of HBV among the five HD centres was 7.5%. The main risk factors were HD centre (26.66%), history of blood transfusion (20%), dental procedure (13.33%) and time duration on HD (6.66%). However, the overall prevalence of HCV among the five HD centres was 19.58%. The main risk factors included HD centre (25.53%), history of blood transfusion (25.53%), dental procedure (10.64%), surgical treatment (6.38%), patients treated abroad (6.38%) and time duration on HD (4.25%). CONCLUSION: The high prevalence of hepatitis viruses among HD patients of northern Pakistan indicates a close relation between HD centres and hepatitis virus transmission. Therefore, preventive control measures are essential to reduce hepatitis transmission in HD centres.

Antibiotic susceptibility profile of bacterial isolates from post-surgical wounds of patients in tertiary care hospitals of Peshawar, Pakistan.

Postoperative wound infections are the infections of the operating site within thirty days after surgery. The infections that develop after surgery are a major problem throughout the world leading to, increased morbidity and mortality. This study was carried out to determine the prevalence of bacterial pathogens causing wound infection in the surgical wards and to determine the antimicrobial sensitivity patterns of the isolated bacteria. A total of 250 wound samples were collected over a period of 6 months from July-December, 2016. The pathogenic bacteria were isolated, identified and their antibiotic susceptibility was determined through disc diffusion method. Among 250 cases, 210 (84%) were culture positive for bacterial pathogens, while 40 (16%) were bacteriologically sterile (Negative). Rate of infection was high in males (55.6%) than females (44.4%). The predominant isolates were E.coli 55 (26.19%), followed by S.aureus 51 (24.28%), Pseudomonas spp. 43(20.47%), S.aureus MRSA 21 (10%), Proteus Marbillis 15 (7.14%), E.coli ESBL producer was 8 (3.81%), Acinetobacter 7 (3.33%) Proteus valgaris 5 (2.38%), b-Streptococci 3 (1.43%) and Klebsella pneumonia were the least, 2 (0.95%). Linezolid, Vancomycin, Amoxycillin, Cefoperazone and Meropenem are the most effective antibiotics for treating post-surgical wound infections.

Prevalence of Hepatitis C virus and its risk factors in blood donors in district Peshawar.

The current study was designed in order to elucidate the most sensitive method for daily practice as well as to evaluate the risk factors for HCV infection associated with blood transfusion in District Peshawar. A total of 1400 healthy volunteer blood donors were tested for Anti-HCV. A questionnaire was used to evaluate the risk factors. Initial testing of all blood samples was done by Immuno Chromatographic Technique (ICT) and confirmed by micro particle enzyme immunoassay (MEIA) and Enzyme Linked Immunosorbent Assay (ELISA). The comparison among ICT, ELISA and MEIA techniques was also evaluated for the purpose of sensitivity. Among 1400 blood donors, 26 (1.85%) cases were found positive for Anti-HCV. These 26 cases were positive on MEIA, 16 individuals were positive on ELISA while 14 were positive on ICT. These 26 cases had different histories of dental treatment (50%), traveled abroad (23.07%), surgery (11.53%), blood transfusion (7.69%) and unknown reason (7.69%). Among all these different histories of dental treatment and blood transfusion were the main risk factors for HCV infection. The results revealed that MEIA is a quick and reliable technique for routine screening of blood donors particularly for controlling the spread of HCV.