Multidrug resistance among uropathogenic clonal group A E. Coli isolates from Pakistani women with uncomplicated urinary tract infections.

OBJECTIVE: Multi-drug resistance (MDR) has notably increased in community acquired uropathogens causing urinary tract infections (UTIs), predominantly Escherichia coli. Uropathogenic E. coli causes 80% of uncomplicated community acquired UTIs, particularly in pre-menopausal women. Considering this high prevalence and the potential to spread antimicrobial resistant genes, the current study was conducted to investigate the presence of clinically important strains of E. coli in Pakistani women having uncomplicated cystitis and pyelonephritis. Women belonging to low-income groups were exclusively included in the study. Seventy-four isolates from urine samples were processed, phylotyped, and screened for the presence of two Single Nucleotide Polymorphisms (SNPs) particularly associated with a clinically important clonal group A of E. coli (CgA) followed by antibiotic susceptibility testing and genome sequence analysis. RESULTS: Phylogroup B2 was most prevalent in patients and 44% of isolates were positive for the presence of CgA specific SNPs in Fumarate hydratase and DNA gyrase subunit B genes. Antibiotic susceptibility testing showed widespread resistance to trimethoprim-sulfamethoxazole and extended-spectrum beta-lactamase production. The infection analysis revealed the phylogroup B2 to be more pathogenic as compared to the other groups. The genome sequence of E. coli strain U17 revealed genes encoding virulence, multidrug resistance, and host colonization mechanisms. CONCLUSIONS: Our research findings not only validate the significant occurrence of multidrug-resistant clonal group A E. coli (CgA) in premenopausal Pakistani women suffering from cystitis and pyelonephritis but also reveal the presence of genes associated withvirulence, and drug efflux pumps. The detection of highly pathogenic, antimicrobial-resistant phylogroup B2 and CgA E. coli strains is likely to help in understanding the epidemiology of the pathogen and may ultimately help to reduce the impact of these strains on human health. Furthermore, the findings of this study will particularly help to reduce the prevalence of uncomplicated UTIs and the cost associated with their treatment in women belonging to low-income groups.

A prospective study on linking diarrheagenic E. coli with stunted childhood growth in relation to gut microbiome.

Stunted growth is an emerging global challenge affecting children under the age of 5 years in low- and middle-income countries. Despite such a high global prevalence of stunting, the mechanism of pathogenesis and the role of associated gut microbiota is poorly understood. The present study was designed to investigate the association of pathogenic strains of E. coli with the residential gut microbiota of stunted growth children. A total of 64 stool sample were collected from children aged ≤ 5 years, and were processed for isolation and molecular characterization of diarrheagenic E. coli. Selected stool samples (n = 39 including three normal controls) were then analysed for microbial community profiling using 16S ribosomal RNA (rRNA) gene sequencing. Furthermore, associations between changes in the microbiota in the presence of different E. coli strains was explored. Pathotyping of the isolated E. coli (n = 64) has shown that 39.68% belonged to one of the five pathotypes of E. coli whilst the remaining ones were non-typeable. Amongst the different pathotypes, EPEC was found to be the most prevalent (52%; n = 13), followed by EAEC (20%; n = 5), EIEC (12%; n = 3), EHEC (8%; n = 2) and ETEC 2 (8%; n = 2). Phylogrouping analysis has shown that majority of the strains belonged to B2 (28.12%). Microbial diversity is shown to be significant and varied when the samples are organized under the recovered phylogroups. Moreover, based on predictive metabolism, the colonization of these strains were found to be significantly associated with energy utilization pathways such as Denovoprine-2 and glyoxylate-by. Differential analysis has shown that Escherichia-Shigella and Enterococcus were altered for the children with stunted growth.

High prevalence of Panton-Valentine Leucocidin (PVL) toxin carrying MRSA and multidrug resistant gram negative bacteria in late onset neonatal sepsis indicate nosocomial spread in a Pakistani tertiary care hospital.

BACKGROUND: Neonatal sepsis has high incidence with significant mortality and morbidity rates in Pakistan. We investigated common etiological patterns of neonatal sepsis at a tertiary care setup. METHODS: 90 pus and blood, gram negative and gram positive bacterial isolates were analyzed for virulence and antibiotic resistance gene profiling using PCR and disc diffusion methods. RESULTS: Staphylococcus aureus showed strong association with neonatal sepsis (43 %) followed by Citrobacter freundii (21 %), Pseudomonas aeruginosa (13 %), Escherichia coli (15 %) and Salmonella enterica (8 %). Molecular typing of E. coli isolates depicted high prevalence of the virulent F and B2 phylogroups, with 4 hypervirulent phylogroup G isolates. 76.9 % S. aureus isolates showed presence of Luk-PV, encoding for Panton-valentine leucocidin (PVL) toxin with majority also carrying MecA gene and classified as methicillin resistant S. aureus (MRSA). ecpA, papC, fimH and traT virulence genes were detected in E. coli and Salmonella isolates. 47 % Citrobacter freundii isolates carried the shiga like toxin SltII B. Antimicrobial resistance profiling depicted common resistance to cephalosporins, beta lactams and fluoroquinolones. CONCLUSION: Presence of PVL carrying MRSA and multidrug resistant gram negative bacteria, all isolated from late onset sepsis neonates indicate a predominant nosocomial transmission pattern which may complicate management of the disease in NICU setups.

Colibactin possessing E. coli isolates in association with colorectal cancer and their genetic diversity among Pakistani population.

Colorectal cancer (CRC) is the third most prevalent cause of tumorigenesis and several pathogenic bacteria have been correlated with aggressive cases of cancer i.e., genotoxin (colibactin) producing Escherichia coli (E. coli). This study was designed to investigate the genetic diversity of clb+clb+ E. coli strains and their association with CRC. Pathogenic E. coli isolates from colorectal biopsies were characterized based on phylotypes, antibiotic resistance pattern, and (Enterobacterial Repetitive Intergenic Consensus Sequence-based Polymerase Chain Reaction) ERIC-PCR. Furthermore, isolates were screened for the presence of the Pks (polyketide synthase) Island specifically targeting colibactin genes A and Q. The selective clb+clb+ isolates were subjected to cytotoxicity assay using Human embryonic kidney (HEK) cell lines. We revealed that 43.47% of the cancer-associated E. coli isolates were from phylogroup B2 comparatively more pathogenic than rest while in the case of healthy controls no isolate was found from B2. Moreover, 90% were found positive for colibactin and pks (polyketide synthase) island, while none of the healthy controls were found positive for colibactin genes. All healthy and cancer-associated isolates were tested against 15 antibiotic agents, we observed that cancer-associated isolates showed a wide range of resistance from 96% against Nalidixic acid to 48% against Doxycycline. Moreover, E. coli isolates were further genotyped using ERIC-PCR, and selected clb+clb+ E. coli isolates were subjected to cytotoxicity assay. We recorded the significant cytotoxic activity of clb+clb+ E. coli phylogroup B2 isolates that might have contributed towards the progression of CRC or dysbiosis of healthy gut microbiota protecting against CRC pathogenesis. Our results revealed a significant p<0.023 association of dietary habits and hygiene p<0.001with CRC. This is the first study to report the prevalence of E. coli phylogroups and the role of colibactin most virulent phylogroup B2 among Pakistani individuals from low socioeconomic setup.

In Silico and In Vitro Analysis of Helicobacter pullorum Type Six Secretory Protein Hcp and Its Role in Bacterial Invasion and Pathogenesis.

Helicobacter pullorum is a human zoonotic pathogen transmitted through poultry where it is associated with vibrionic hepatitis and colitis. Hemolysin co-regulated protein (Hcp) is an important structural as well as effector protein of type six secretory system; however, its role in H. pullorum invasion and pathogenesis has not been elucidated. In this study, we predicted the Helicobacter pullorum Hcp (HpuHcp) structure and identified Campylobacter jejuni Hcp (CjHcp) as its nearest homologue. Analysis of the predicted structure shows several common bacterial Hcp motifs like Protein kinase C phosphorylation site, Casein kinase II phosphorylation site, N-myristoylation site, cAMP-and cCGMP-dependent protein kinase phosphorylation site, N-glycosylation site. The presence of unique microbodies C-terminal targeting signal domain was present in HpuHcp which was seen for the first time in CjHcp. This could indicate that Hcp is a structural protein as well as a secretory protein. Moreover, the presence of a deamidase domain, similar to the tecA of Burkholderia cenocepacia an opportunistic pathogen, may help in bacterial internalization as it depolymerises the membranous actin by deamidation of the host cell Rho GTPases cdc42 and Rac1, which was supported by increased invasion of hepatocytes by Hcp-positive isolates.

Capillary-Driven Flow Microfluidics Combined with Smartphone Detection: An Emerging Tool for Point-of-Care Diagnostics.

Point-of-care (POC) or near-patient testing allows clinicians to accurately achieve real-time diagnostic results performed at or near to the patient site. The outlook of POC devices is to provide quicker analyses that can lead to well-informed clinical decisions and hence improve the health of patients at the point-of-need. Microfluidics plays an important role in the development of POC devices. However, requirements of handling expertise, pumping systems and complex fluidic controls make the technology unaffordable to the current healthcare systems in the world. In recent years, capillary-driven flow microfluidics has emerged as an attractive microfluidic-based technology to overcome these limitations by offering robust, cost-effective and simple-to-operate devices. The internal wall of the microchannels can be pre-coated with reagents, and by merely dipping the device into the patient sample, the sample can be loaded into the microchannel driven by capillary forces and can be detected via handheld or smartphone-based detectors. The capabilities of capillary-driven flow devices have not been fully exploited in developing POC diagnostics, especially for antimicrobial resistance studies in clinical settings. The purpose of this review is to open up this field of microfluidics to the ever-expanding microfluidic-based scientific community.

HriGFP Novel Flourescent Protein: Expression and Applications.

Biosensors based on microbial cells have been developed to monitor environmental pollutants. These biosensors serve as inexpensive and convenient alternatives to the conventional lab based instrumental analysis of environmental pollutants. Small monomeric naturally occurring fluorescent proteins (fp) can be exploited by converting them as small biosensing devices for biomedical and environmental applications. Moreover, they can withstand exposure to denaturants, high temperature, and a wide pH range variation. The current study employs newly identified novel fluorescent protein HriGFP from Hydnophora rigida to detect environmental contaminants like heavy metals and organo-phosphorous (pesticide) compounds such as methyl parathion. The HriGFP was initially tested or its expression in bacterial systems (Gram positive and Gram negative) and later on for its biosensing capability in E coli (BL21DE3) for detection of heavy metals and methyl parathion was evaluated. Our results indicated the discrete and stable expression of HriGFP and a profound fluorescent quenching were observed in the presence of heavy metals (Hg, Cu, As) and methyl parathion. Structural analysis revealed heavy metal ions binding to HriGFP via amino acid residues. In-silico-analysis further revealed strong interaction via hydrogen bonds between methyl parathion phosphate oxygen atoms and the amino group of Arg119 of HriGFP. This study implies that HriGFP can act as a biosensor for detecting harmful carcinogenic pesticide like methyl parathion in water resources in the vicinity of heavily pesticide impregnated agricultural lands and heavy metal contaminated water bodies around industrial areas.

Identification of novel bacterial urease inhibitors through molecular shape and structure based virtual screening approaches.

The enzyme urease is an essential colonizing factor of the notorious carcinogenic pathogen Helicobacter pylori (H. pylori), conferring acid resistance to the bacterium. Recently, antibiotic resistant strains have emerged globally with little to no alternative treatment available. In this study we propose novel urease inhibitors capable of controlling infection by H. pylori and other pathogenic bacteria. We employed hierarchal computational approaches to screen new urease inhibitors from commercial chemical databases followed by in vitro anti-urease assays. Initially ROCS shape-based screening was performed using o-chloro-hippurohydroxamic acid followed by molecular docking studies. Out of 1.83 million compounds, 1700 compounds were retrieved based on having a ROCS Tanimoto combo score in the range of values from 1.216 to 1.679. These compounds were further screened using molecular docking simulations and the 100 top ranked compounds were selected based on their Glide score. After structural classification of the top ranked compounds, eight compounds were selected and purchased for biological assays. The plausible binding modes of the most active compounds were also confirmed using molecular dynamics (MD) simulations. Compounds 1, 2 and 3 demonstrated good urease inhibitory properties (IC50 = 0.32, 0.68 and 0.42 µM) compared to the other compounds. Enzyme kinetic studies revealed that compounds 1 and 3 are competitive inhibitors while 2 is a mixed type inhibitor of the urease enzyme. Cell based urease inhibition and MTT assay showed that these compounds blocked H. pylori urease activity, affecting bacterial growth and acid tolerance.

Transmission of multidrug-resistant Campylobacter jejuni to children from different sources in Pakistan.

OBJECTIVES: Due to the rapid emergence of multidrug-resistant isolates, Campylobacter jejuni (C. jejuni) has been classified as a member of the priority pathogens group. This study aimed to determine the prevalence, antibiotic resistance patterns and source tracking of clinical C. jejuni isolates from paediatric diarrhoeal patients in Pakistan. METHODS: A total of 150 stool samples from children were processed for the presence of C. jejuni using culture, biochemical tests and species-specific PCR. Antibiotic susceptibility of the isolates was determined by the disc diffusion method, and metallo-ß-lactamase (MBL) producers were detected using gene-specific PCR. Source tracking was performed using source-predictive PCR. RESULTS: C. jejuni was present in 54.6% of the processed samples. More than 80% of the isolated strains were resistant to seven of 12 tested antibiotics. High levels of susceptibility were observed against imipenem (12.2%) and TGC (9.7%). Six isolates (7.3%) were MBL producers and positive for at least one of the five MBL genes. Source tracking showed that 57.3% of the isolates belonged to livestock-associated clusters (C1-C6) and 42.8% were assigned to non-livestock/environmental clusters (C7-C9). Isolates belonging to livestock clusters had a high Multiple Antibiotic Resistance (MAR) index (P < 0.001) as compared with non-livestock. CONCLUSION: A high prevalence of multidrug-resistant C. jejuni among paediatric diarrhoeal patients was observed. Moreover, the association of these isolates to livestock clades suggests transmission to human populations via the food chain. The presence of imipenem-resistant MBL-producing C. jejuni can lead to serious public health concerns.

Magnitude of Rotavirus A and Campylobacter jejuni infections in children with diarrhea in Twin cities of Rawalpindi and Islamabad, Pakistan.

BACKGROUND: Acute diarrhea is a leading cause of morbidity and mortality in children particularly in developing countries of Asia and Africa. The present study was conducted to detect the two most important pathogens, rotavirus and Campylobacter Jejuni in children suffering with diarrhea in Rawalpindi and Islamabad, Pakistan in 2014. The clinical and epidemiological aspects of the disease were also investigated. METHODS: A total of 500 stool samples were collected from children presented with clinical signs and symptoms of acute diarrhea. The samples were initially screened for the presence of rotavirus A (RVA) via ELISA (Enzyme-linked immunosorbent assay) and RT-PCR (Reverse Transcriptase PCR) and then were analysed for C. jejuni by using species specific PCR assay. RESULTS: The detection rate of RVA was 26.4% (132/500) while, Campylobacter was detected in 52% (260/500) of samples with C. jejuni accounted for 48.2% (241/500) of all study cases. Co-infection of C. jejuni with RVA was identified in 21.8% of all cases. Children with RVA and C. jejuni co-infection showed a higher probability (p = 0.01) to be dehydrated. A significant association (p = 0.02) was found between C. jejuni positive status and fever in children. The median age of children with both RVA and C. jejuni infection was 6-11 months. The RVA detection rate was high in winter months of the year while, C. jejuni infections were documented high in summer over 1 year study period. CONCLUSIONS: The overall results have demonstrated the high prevalence of C. jejuni in Rawalpindi, Islamabad, Pakistan in 2014. The results of present study will not only help to calculate disease burden caused by C. jejuni and rotavirus but also will provide critical information to health authorities in planning public health care strategies against these pathogens.

Variation in antibiotic susceptibility and presence of type VI secretion system (T6SS) in Campylobacter jejuni isolates from various sources.

Campylobacter jejuni is a major cause of infectious diarrhea in humans. The bacterium can be transmitted through contaminated poultry meat and waste water. We report the presence of C. jejuni from potential transmission sources including egg shells, poultry waste, waste water and migratory bird droppings with a prevalence rate of 78%, 66%, 86% and 70% respectively. Antibiotic resistance profile showed high number of isolates resistant to multiple antibiotics including 4th generation cephalosporins. C. jejuni isolates were further screened for presence of T6SS, an important virulence factor. None of the C. jejuni isolates from migratory birds carried a T6SS, whereas highest prevalence of T6SS isolates was observed in waste water samples, followed by poultry waste and egg shells. To determine virulence potential of the isolates, hemolytic activity of isolates was compared. Although variation in hemolytic potential between isolates from different sources was noted, higher hemolytic activity was observed for isolates possessing hcp, a T6SS gene. Furthermore, presence of T6SS affords the bacterium some survival advantage when compared to T6SS competent Helicobacter pullorum which occupies the same niche. Taken together our findings indicate that C. jejuni with T6SS have a fitness advantage increasing their isolation frequency from waste water and poultry waste.

Prevalence and role of Type six secretion system in pathogenesis of emerging zoonotic pathogen Helicobacter pullorum from retail poultry.

Helicobacter pullorum is an emerging zoonotic pathogen that causes gastroenteritis in chickens and inflammatory bowel disease in humans ingesting contaminated meat. However, the mechanism by which the bacterium causes disease is unclear. Type six secretion system (T6SS) plays a major role in bacterial pathogenesis and adaptation. Haemolysin coregulated protein (Hcp) plays a central role in the structure of the T6SS pilus and acts as effector protein in certain bacteria. In this study, H. pullorum isolated from 156 caecal samples of broiler chickens was screened for the presence of T6SS Hcp gene via PCR amplification. 30.7% of caecal and 18.3% of liver samples tested positive for presence of H. pullorum. From these positive samples, 29.7% possessed the T6SS gene. In bacterial co-culture experiments, significant loss of viability (81.6-39.1%) was observed for H. pullorum-infected hepatocytes and presence of Hcp did not contribute to the loss of cell viability. Nevertheless, infection of erythrocytes with Hcp-positive isolates was associated with greater haemolytic activity compared to infection with Hcp-negative isolates. Therefore, presence of T6SS could be indicative of virulent strains meriting further studies to characterize this virulence factor in H. pullorum infection.

Visible light sensitive Ag/TiO2/graphene composite as a potential coating material for control of Campylobacter jejuni.

Infectious diarrhea caused by the food borne pathogen, Campylobacter jejuni, is a major threat to public health worldwide leads high incidence of child mortality each year. In the present study, hydrothermal synthesis of Silver-Graphene-TiO2 nanocomposites along with TiO2, TiO2-Graphene and TiO2-silver nanocomposites was done and the samples were characterized using X-ray diffraction (XRD), tunneling electron microscopy (TEM) and UV-Vis Spectroscopy. Effect of silver and graphene addition on the broad spectrum antibacterial ability of TiO2 was studied under visible light. Moreover, the effects on bacterial survival, membrane integrity, cellular motiltiy and biofilm formation of C. jejuni were also evaluated. A synergetic effect of silver and graphene on Silver-Graphene-TiO2 nanocomposites was observed as indicated by its increased visible light sensitivity and enhanced antibacterial activity under visible light compared to its parent derivatives. Silver-Graphene-TiO2 composites effectively reduced growth and caused leakage of protein and DNA from C. jejuni cell. Atomic Force Microscopy was used to confirm bacterial cell damage. Besides, it also reduced motillity, hydrophobicity and autoaggregation of C. jejuni and showed excellent inhibition of biofilm formation. Furthermore, no significant cytotoxicity of synthesized nanoparticles was observed in human cell lines. We propose that Silver-Graphene-TiO2 composites can be used as effective antimicrobial agents to control the spread of C. jejuni by preventing both bacterial growth and biofilm formation.

Structural basis for the pathogenesis of Campylobacter jejuni Hcp1, a structural and effector protein of the Type VI Secretion System.

The Type VI Secretion System (T6SS) provides enhanced virulence to Campylobacter jejuni and has been associated with a high incidence of bloody diarrhea. The hemolysin-coregulated protein (Hcp)-the hallmark of the T6SS-can act as a structural and effector protein. Unlike other T6SS-harboring bacteria, which possess multiple Hcp proteins each performing different functions, C. jejuni possesses only one Hcp protein, and its structural and functional role has not been elucidated previously. Here, we report the structure and functional studies of Hcp from C. jejuni. We found similarities between the hexameric ring structure of Hcp-Cj and that of Hcp3 from Pseudomonas aeruginosa. Through functional studies, we showed two roles for Hcp-Cj that is, in cytotoxicity toward HepG2 cells and in biofilm formation in C. jejuni. In structure-based mutational analyses, we showed that an Arg-to-Ala mutation at position 30 within the extended loop results in a significant decrease in cytotoxicity, suggesting a role for this loop in binding to the host cell. However, this mutation does not affect its biofilm formation function. Collectively, this study supports the dual role of Hcp-Cj as a structural and effector protein in C. jejuni.

Galleria mellonella is low cost and suitable surrogate host for studying virulence of human pathogenic Vibrio cholerae.

Vibrio cholerae causes a severe diarrheal disease affecting millions of people worldwide, particularly in low income countries. V. cholerae successfully persist in aquatic environment and its pathogenic strains results in sever enteric disease in humans. This dual life style contributes towards its better survival and persistence inside host gut and in the environment. Alternative animal replacement models are of great value in studying host-pathogen interaction and for quick screening of various pathogenic strains. One such model is Galleria mellonella, a wax moth which has a complex innate immune system and here we investigate its suitability as a model for clinical human isolates of O1 El TOR, Ogawa serotype belonging to two genetically distinct subclades found in Pakistan (PSC-1 and PSC-2). We demonstrate that the PSC-2 strain D59 frequently isolated from inland areas, was more virulent than PSC-1 strain K7 mainly isolated from coastal areas (p=0.0001). In addition, we compared the relative biofilm capability of the representative strains as indicators of their survival and persistence in the environment and K7 showed enhanced biofilm forming capabilities (p=0.004). Finally we present the annotated genomes of the strains D59 and K7, and compared them with the reference strain N16961.

Resistance patterns of diversified phylogroups of Escherichia coli associated with mothers having history of preterm births in Pakistan.

OBJECTIVE: Urinary tract infections (UTIs) are caused by extraintestinal pathogenic Escherichia coli (ExPEC), and are one of the key predictors of preterm births. In the light of this fact, present study was conducted to determine the predominant Escherichia coli (E. coli) phylotypes and their associated antibiotic susceptibility patterns, isolated from pregnant mothers with the history of preterm births. METHODS: Forty seven E. coli strains were isolated out of a total of 80 urine samples of pregnant women. The isolates were phylotyped and further screened for the presence of Clonal group A. Moreover, Antimicrobial susceptibility testing and screening for Extended Spectrum Beta Lactamase (ESBL) producing strains were also performed. RESULTS: Among the 47 isolates, phylogroup B2 was found to be highly prevalent (45%), followed by group D (23%), B1 (10.64%), A (6.38%), E (6.38%), cryptic clade I (4.25%) and F (2.13%). Two isolates belonged to CgA and 41 (87.23%) isolates were found to be multidrug-resistant. Out of nine antibiotics tested in the study, the isolates displayed high resistance to Ampicillin (82.6%), Sulphamethoxazole (65.22%), Nalidixic acid (60.87%), Sulphamethoxazole-Trimethoprim, Doxycycline and Erythromycin (56.52% each). In total, 8 (17.02%) of the isolates were found to be ESBL positive. CONCLUSIONS: The prevalence of infections caused by virulent and highly drug resistant E. coli isolates constitute a risk of developing preterm birth complications in pregnant women and requires the selection of appropriate antibiotics for the treatment of infections caused during pregnancy.

Characterization of enteropathogenic Escherichia coli of clinical origin from the pediatric population in Pakistan.

BACKGROUND: Enteropathogenic Escherichia coli (EPEC) is one of the leading causes of watery diarrhea among children. METHODS: In this study EPEC isolates from the pediatric population of Pakistan (2010-2012) were subjected to phylotyping, antibiotic susceptibility, extended-spectrum beta-lactamase (ESBL) profiling and evaluation of one representative strain from each panel of phylotypesin Galleria mellonella, infection model. RESULTS: A total of 46/225 (20.4%) stool samples were positive for EPEC. Isolates mainly belong to D phylogroup (18, 39%) followed by nontypeable (10, 22%), B1 (9, 20%), B2 (8, 17%) and A (1, 2%). High resistance was observed for ampicillin (42, 91%), erythromycin (41, 89%), cefaclor (37, 80%), trimethoprim/sulfamethoxazole (36, 78%), tetracycline (36, 78%). Among nalidixic acid resistant isolates 13 (28%) showed presence of single nucleotide polymorphism (SNP) in parC (C330-T330) whereas 1 (2%) isolate showed gyrB (A660-T660) SNP. Furthermore, 27 (59%) isolates were ESBL producers. Representative isolates of phlyotypes A and B2 showed enhance killing of G. mellonella compared to ones belonging to phylotypes B1 and D. CONCLUSIONS: Non-typeable EPEC strains were frequently observed. ESBL production in ESBL producers was found to be plasmid mediated. No significant association of antibiotic resistance profile with specific phylogroup of EPEC was found, however G. mellonella infection model differentiated representative phylotypes.

Antibacterial, Structural and Optical Characterization of Mechano-Chemically Prepared ZnO Nanoparticles.

Structural investigations, optical properties and antibacterial performance of the pure Zinc Oxide (ZnO) nanoparticles (NPs) synthesized by mechano-chemical method are presented. The morphology, dimensions and crystallinity of the ZnO NPs were controlled by tweaking the mechanical agitation of the mixture and subsequent thermal treatment. ZnO nanoparticles in small (< 20 nm) dimensions with spherical morphology and narrow size distribution were successfully obtained after treating the mechano-chemically prepared samples at 250°C. However, higher temperature treatments produced larger particles. TEM, XRD and UV-Vis spectroscopy results suggested crystalline and phase pure ZnO. The NPs demonstrated promising antibacterial activity against Gram negative foodborne and waterborne bacterial pathogens i.e. Enteropathogenic E. coli (EPEC), Campylobacter jejuni and Vibrio cholerae as well as Gram positive methicillin resistant Staphylococcus aureus (MRSA), thus potential for medical applications. Scanning electron microscopy and survival assay indicated that most probably ZnO nanoparticles cause changes in cellular morphology which eventually causes bacterial cell death.

Draft Genome Sequence of the Enteropathogenic Bacterium Campylobacter jejuni Strain cj255.

The enteropathogen Campylobacter jejuni is a global health disaster, being one of the leading causes of bacterial gastroenteritis. Here, we present the draft genome sequence of C. jejuni strain cj255, isolated from a chicken source in Islamabad, Pakistan. The draft genome sequence will aid in epidemiological studies and quarantine of this broad-host-range pathogen.