Complete genome sequence of Escherichia phage iGC_PHA_EC001.

Antimicrobial resistance (AMR) in bacteria poses a global health emergency due to limited treatment options. Here, we report a lytic bacteriophage belonging to Stephanstirmvirinae family against an AMR Escherichia coli (ST2089). Escherichia phage iGC_PHA_EC001 is of genus Phapecoctavirus and 148,445 bp in length, encoding 269 predicted protein-coding sequences and 10 tRNAs. The phage encodes two lytic proteins containing phage_lysozyme (PF00959.22) and cell wall hydrolase_2 (PF07486.15) as catalytic domains, respectively.

Whole genomes from bacteria collected at diagnostic units around the world 2020.

The Two Weeks in the World research project has resulted in a dataset of 3087 clinically relevant bacterial genomes with pertaining metadata, collected from 59 diagnostic units in 35 countries around the world during 2020. A relational database is available with metadata and summary data from selected bioinformatic analysis, such as species prediction and identification of acquired resistance genes.

Characterization of lactic acid bacteria isolated from street pickles of Dhaka, Bangladesh.

Traditionally fermented pickles are a popular street food in Bangladesh famous for their unique flavors and health benefits. Pickles are often prepared by fermentation using lactic acid bacteria (LAB) that can act as probiotics. The study was aimed to isolate and characterize lactic acid bacteria from pickle samples collected from streets of Dhaka city, as well as assess the microbial quality of pickles for food safety. A total of 30 pickle samples of different kinds were collected from streets of Dhaka city. Isolation and identification were conducted using conventional cultural and biochemical tests, followed by molecular confirmation of identity. Antibiotic susceptibility of isolates was investigated against 7 antibiotics of different groups. Antimicrobial activity of LAB isolates was analyzed by well-diffusion assay and phenotypic enterocin activity assay. Physiological characterizations of LAB were performed to determine their tolerance to temperature, salt, pH, bile, carbohydrate fermentation pattern, proteolytic activity and biofilm formation. Fifty isolates were obtained from pickle samples, of which 18% was identified as LAB, including Enterococcus faecalis (6) and Enterococcus faecium (3). The rest included S. aureus (18), E. coli (11), Klebsiella spp. (5), Salmonella (3), Shigella (3) and Pseudomonas aeruginosa (1). Antibiotic resistance pattern revealed higher occurrence of resistance against azithromycin among the non-LAB isolates, but none of the LAB isolates were found to resist any of the antibiotics used. Antimicrobial activity of LAB isolates was not observed against the foodborne isolates. All LAB isolates fermented a wide range of carbohydrates and showed adequate tolerance to salt, pH, temperature and bile. Out of 9 isolates, 5 displayed proteolytic activity, and 6 were found as strong biofilm producer. These results suggest that although the LAB isolates from street pickles collected from Dhaka does not have antimicrobial activities, they still have potential to be used as probiotics. It also shows high occurrence of antibiotic resistant foodborne pathogens in pickles, indicating that consumption of such street food can be serious health hazard.

Detecting Enteric Pathogens in Low-Risk Drinking Water in Dhaka, Bangladesh: An Assessment of the WHO Water Safety Categories.

The microbiological quality of water is usually assessed by fecal coliform bacteria, and the presence of E. coli as an indicator of fecal contamination is widely recommended by international guidelines. This study aimed to assess the prevalence of diarrheagenic pathogens, in both public and personal domain water sources and examine the reliance on the WHO drinking water risk assessment guidelines. This study was conducted in a low-income urban community in Dhaka, Bangladesh between September 2014 and October 2015. Polymerase chain reaction (PCR) was used to detect the marker and virulence genes of Escherichia coli, Vibrio cholerae, Salmonella species, and Campylobacter species, and the culture method was employed for the quantitative assessment of E. coli. According to the WHO guidelines, 48% of the public domain source water and 21% of the personal domain point-of-drinking water were classified in the low-risk group, i.e., 0 CFU of E. coli/100 mL. However, when using PCR, we detected pathogens in 39% (14/36) of the point-of-drinking water samples and 65% (74/114) of the public domain water source samples classified in the low-risk group. Our study showed that relying solely on E. coli detection as a measure of water quality may overlook the presence of other pathogens in the drinking water. In addition to the culture-based method, the detection of virulence genes by PCR should also be considered to add more scrutiny to the detection of diverse types of pathogens.

Introduction of the Personal Domain in Water Sanitation and Hygiene (WASH), a New Approach to Identify Missing Health Impacts.

The water sanitation and hygiene (WASH) sector has provided beneficiaries in low and middle-income countries with latrines and clean water for decades. However, we still need good evidence documenting the expected health impact. This paper investigates why we lack this evidence and ways to move forward. Using mTEC agar, we monitored E. coli contamination on selected "hotspot" surfaces within the kitchen environments of 32 low-income households in Dhaka, Bangladesh, every six weeks for two years. Despite being washed, the highest average contamination was found on food plates, at 253 cfu/10 cm2, followed by cutting knives, with 240 cfu/10 cm2. The drinking vessel surfaces and the latrine doorknobs had the lowest contaminations, with E. coli means of 167 and 73 cfu/10 cm2, respectively. These findings imply a need to measure an individual's pathogen exposure as close to the mouth as possible to estimate the true pathogen exposure. The paper proposes introducing the new "personal domain"-the point of consumption-as the physical sphere in which WASH interventions should be assessed. With this approach, we can observe and quantify the different pathogen exposure routes and, with this, further improve WASH interventions.

A coordinated approach for managing polypharmacy among children with medical complexity: rationale and design of the Pediatric Medication Therapy Management (pMTM) randomized controlled trial.

BACKGROUND: Children with medical complexity (CMC) often rely upon the use of multiple medications to sustain quality of life and control substantial symptom burden. Pediatric polypharmacy (≥ 5 concurrent medications) is prevalent and increases the risk of medication-related problems (MRPs). Although MRPs are associated with pediatric morbidity and healthcare utilization, polypharmacy is infrequently assessed during routine clinical care for CMC. The aim of this randomized controlled trial is to determine if a structured pharmacist-led Pediatric Medication Therapy Management (pMTM) intervention reduces MRP counts, as well as the secondary outcomes of symptom burden and acute healthcare utilization. METHODS: This is a hybrid type 2 randomized controlled trial assessing the effectiveness of pMTM compared to usual care in a large, patient-centered medical home for CMC. Eligible patients include all children ages 2-18 years old, with ≥ 1 complex chronic condition, and with ≥ 5 active medications, as well as their English-speaking primary caregivers. Child participants and their primary parental caregivers will be randomized to pMTM or usual care before a non-acute primary care visit and followed for 90 days. Using generalized linear models, the overall effectiveness of the intervention will be evaluated using total MRP counts at 90 days following pMTM intervention or usual care visit. Following attrition, a total of 296 CMC will contribute measurements at 90 days, which provides > 90% power to detect a clinically significant 1.0 reduction in total MRPs with an alpha level of 0.05. Secondary outcomes include Parent-Reported Outcomes of Symptoms (PRO-Sx) symptom burden scores and acute healthcare visit counts. Program replication costs will be assessed using time-driven activity-based scoring. DISCUSSION: This pMTM trial aims to test hypotheses that a patient-centered medication optimization intervention delivered by pediatric pharmacists will result in lower MRP counts, stable or improved symptom burdens, and fewer cumulative acute healthcare encounters at 90 days following pMTM compared to usual care. The results of this trial will be used to quantify medication-related outcomes, safety, and value for a high-utilization group of CMC, and outcomes may elucidate the role of integrated pharmacist services as a key component of outpatient complex care programs for this priority pediatric population. TRIAL REGISTRATION: This trial was prospectively registered at clinicaltrials.gov (NCT05761847) on Feb 25, 2023.

Abundance and biofilm formation capability of Vibrio cholerae in aquatic environment with an emphasis on Hilsha fish (Tenualosa ilisha).

The potentially deadly and sporadic diarrhea-causing agent, Vibrio cholerae, is present in a great number in the freshwater aquatic environment and can be transmitted to humans by different aquatic organisms. In the perspective of Bangladesh, an anadromous fish species Hilsha (Tenualosa ilisha) can act as a transmission vehicle of V. cholerae from the aquatic to the household kitchen environment. The present study was carried out to investigate the presence of V. cholerae in the aquatic habitat of Bangladesh with a major emphasis on freshly caught Hilsha fish, along with river water and plankton samples from the fish capture site. The study also detected the biofilm formation capability of V. cholerae within Hilsha fish that might help the transmission and persistence of the pathogen in aquatic habitat. Twenty out of 65 freshly caught fish (30.8%) and 1 out of 15 water samples (6.67%) showed the presence of V. cholerae and none of the plankton samples were positive for V. cholerae. The isolated strains were identified as non-O1 and non-O139 serogroups of V. cholerae and contain some major toxin and virulence genes. A few strains showed cellular cytotoxicity on the HeLa cell line. All strains were able to form biofilm on the microtiter plate and the detection of three genes related to biofilm formation (vpsA, vpsL, and vpsR) were also assayed using qPCR. In this study, the in vitro biofilm formation ability of the isolated strains may indicate the long-term persistence of V. cholerae in different parts of Hilsha fish. The abundance of V. cholerae only in freshly caught Hilsha fish and the absence of the pathogen in the surrounding aquatic environment could stipulate the role of Hilsha fish as one of the major transmission routes of V. cholerae from the freshwater aquatic environment of Bangladesh to the household kitchen environment.

Escherichia coli Ingested via Food May Overshadow the Positive Effects of Clean Drinking Water: An Example from Dhaka.

The minimal health impact observed in large-scale water sanitation and hygiene (WASH) intervention studies motivated us to investigate the contribution of contaminated food and drinking water to the total daily Escherichia coli load ingested by the average adult in a low-income, urban area. Leftover food (food left at room temperature for more than 6 hours) from 32 households was collected eight times at 6-week intervals in 2014-2015 in the low-income area of Arichpur, Dhaka, Bangladesh. In total, 117 samples were obtained from four food types: fish, lentils, rice, and vegetables, which comprise approximately 85.2% of the average adult's personal daily food consumption. Samples were analyzed for E. coli using selective chromogenic media. For an average adult, the daily consumption of the four food types at mean contamination levels of E. coli can contribute 4.45 log colony-forming units (cfu)/day (95% confidence interval 4.06-4.84). Drinking water quality was measured 211 times at the point of drinking, with a mean, median, and maximum contamination of 1.9, 1.2, and 2.82 log E. coli cfu/100 mL, respectively. If the typical adult in Arichpur were able to drink water with 0 E. coli cfu/100 mL, it would only remove < 5.2% of the total E. coli ingested per day with a mean-contaminated diet. These approximations may suggest why insignificant effects have been observed for water quality interventions in similar, low-hygiene settings. In Arichpur, the E. coli contribution from drinking water to the total E. coli load was insufficient to exert a substantial effect.

Is It Human or Animal? The Origin of Pathogenic E. coli in the Drinking Water of a Low-Income Urban Community in Bangladesh.

This study aimed to investigate the origin of diverse pathotypes of E. coli, isolated from communal water sources and from the actual drinking water vessel at the point-of-drinking inside households in a low-income urban community in Arichpur, Dhaka, Bangladesh, using a polymerase chain reaction (PCR). Forty-six percent (57/125, CI 95%: 41-58) of the isolates in the point-of-drinking water and 53% (55/103, CI 95%: 45-64) of the isolates in the source water were diarrheagenic E. coli. Among the pathotypes, enterotoxigenic E. coli (ETEC) was the most common, 81% (46/57) of ETEC was found in the point-of-drinking water and 87% (48/55) was found in the communal source water. Phylogenetic group B1, which is predominant in animals, was the most frequently found isolate in both the point-of-drinking water (50%, 91/181) and in the source (50%, 89/180) water. The phylogenetic subgroup B23, usually of human origin, was more common in the point-of-drinking water (65%, 13/20) than in the source water (35%, 7/20). Our findings suggest that non-human mammals and birds played a vital role in fecal contamination for both the source and point-of-drinking water. Addressing human sanitation without a consideration of fecal contamination from livestock sources will not be enough to prevent drinking-water contamination and thus will persist as a greater contributor to diarrheal pathogens.

Investigation of the Domestic Reservoirs of Diarrheagenic Escherichia coli in Diarrhea Case Households of Urban Bangladesh.

This study collected rectal swabs from diarrheal patients and in-house environmental samples from low-income households in Dhaka City, Bangladesh, over a 4-month period and investigated these to determine the domestic transmission pathways of Escherichia coli-associated diarrhea. The environmental samples included swabs from four frequently touched surfaces, drinking water and food. Both the rectal swabs and environmental samples were examined for virulence genes characteristic of diarrheagenic E. coli pathotypes by PCR. In addition, each sample was cultured for E. coli, and the strains were analyzed for virulence profile and subjected to multilocus sequence typing (MLST). The results showed that 31% (73 of 233) of all samples including rectal swabs and household samples were positive for one or more of the diarrheagenic E. coli virulence factors. PCR analyses showed that 28% (10/36) of the rectal swabs, 43% (58/136) of household swabs, 9% (3/32) of the food, and 7% (2/29) of the water samples were positive for various virulence genes. 6 Out of the 36 rectal swab samples and associated household samples were shown to have similar E. coli pathotypic genes, and the drinking vessel surface was identified as the major source of contamination. EAEC and CTEC were the most commonly identified pathotypes in the cultured isolates. The phylogenetic tree constructed by MLST data showed that the diarrheagenic isolates were clustered in several diversified lineages. This study supports the hypothesis that there are high-risk hotspots, particularly those surfaces associated with food consumption, for diarrheagenic E. coli contamination within the household environments of Bangladesh.

Historical and contemporary views on cholera transmission: are we repeating past discussions? Can lessons learned from cholera be applied to COVID-19?

Cholera, a devastating diarrheal disease that caused several global pandemics in the last centuries, may share some similarities with the new COVID-19. Cholera has affected many populations in history and still remains a significant burden in developing countries. The main transmission route was thought to be predominantly through contaminated drinking water. However, revisiting the historical data collected during the Copenhagen 1853 cholera outbreak allowed us to re-evaluate the role of drinking-water transmission in a city-wide outbreak and reconsider some critical transmission routes, which have been neglected since the time of John Snow. Recent empirical and cohort data from Bangladesh also strengthened the dynamic potentiality of other transmission routes (food, fomite, fish, flies) for transmitting cholera. Analyzing this particular nature of the cholera disease transmission, this paper will describe how the pattern of transmission routes are similar to COVID-19 and how the method of revisiting old data can be used for further exploration of new and known diseases.

Water usage, hygiene and diarrhea in low-income urban communities-A mixed method prospective longitudinal study.

Epidemiological studies considered water use and hygiene practices as central risk factors for diarrhea. Few studies focused on independent association of water quantity with diarrheal diseases. This study aimed to describe the methodological protocol that adapted multidisciplinary and mixed-method research approach to assess how water usage including water quantity influences the attributable risk for diarrhea in a low-income urban community in Bangladesh. The quantitative, anthropological and microbiological approaches were threaded together to provide a greater understanding of the infrastructural, behavioral and microbial interactions to fathom the dimensions of fecal oral transmission pathways within the households. The use of the 'Choleraphone' (i.e. a mobile phone based real time diarrheal reporting system) was a contemporary approach intended to cut down on resources, reduce research fatigue and provide more accurate data compared to the 'gold standard' (i.e. visiting a household of diarrhea cases within 48 hours) for measuring diarrhea incidence. Development of methods to measure water quantity using qualitative and quantitative approach within a setting where meter water connection is rare was another unique feature of this protocol. This protocol provided guidance and insight on how multiple methods of different disciplines can be combined to enrich understanding of waterborne diseases.

Complete genome and plasmid sequence of a novel Bacillus sp. BD59S, a parasporal protein synthesizing bacterium.

Parasporal crystalline inclusion proteins of some Bacillus spp. are of paramount importance due to their insecticidal, nematocidal, and cancer cell killing capabilities. Here, we present a brief report of the complete genome sequence of Bacillus sp. BD59S, a bacterium that produced HeLa cell-killing parasporal crystalline inclusion proteins. From genome sequencing and assembly, we found that the bacterium has one circular chromosome and two large plasmids, pBTBD59S1 and pBTBD59S2. The size of the chromosome is 5283,933 bp with a 35.4% GC content, consisting of 5938 genes and 5550 protein-coding sequences (CDSs), 25 complete rRNAs (5S, 16S, 23S), 98 tRNAs, 5 ncRNAs, 260 pseudo-genes, and 356 subsystems. Complete plasmid sequence of pBTBD59S1 comprises a total size of 162,149 bp with 33.4% GC content, 192 CDSs, and 13 subsystems. The other plasmid pBTBD59S2, is 199,209 bp long with 32.9% GC content, 179 CDSs, and 11 subsystems. Analyses by NCBI microbial genome BLAST, phylogenetic genome tree, and BLAST ring image generator (BRIG) revealed that BD59S belongs to Bacillus cereus group, and is more close to B. thuringiensis. Further, the strain possesses 57.04 kDa and 54.42 kDa Cry protein-coding genes, which show significant similarities with cancer cell-killing parasporin proteins of B. thuringiensis strains.

Evaluation of adaptive low cost solar water pasteurization device for providing safe potable water in rural households.

This study was conducted to evaluate the effectiveness of a simplified, low cost, pasteurization device in inactivating the diarrheal pathogens present in pond/lake/river water in order to provide safe potable water to people living in the rural areas of low resource countries. In this process, water in polyethylene bags was exposed to sunshine, where UV radiation emissions and heat absorption from the sunshine occurs simultaneously, and maintaining the heating at <60 °C, and minimum UV radiation emissions of 996.2 W/m2 for approximately 30 minutes was found enough to inactivate diarrheal pathogens in water. The synergistic effect of heat, UV radiation emission and holding time causes the destruction of diarrheal pathogens. However, the performance of the device depends on the thickness of the insulation and the air gap between polyethylene bags. Regardless of sample sources, the highest population reduction of Escherichia coli observed in the bacterial challenge study was 6.8 ± 0.4 log CFU/ml. The physicochemical properties were found acceptable compared with USEPA potable water quality except turbidity, which is acceptable according to the BDS standard, and the shelf-life study results demonstrated that 6 months' storage of pasteurization device-treated water at room temperature is possible without compromising water quality. Therefore, this simplified pasteurization device could be useful in potable water-scarce areas of the world.

A Comparative Analysis of Vibrio cholerae Contamination in Point-of-Drinking and Source Water in a Low-Income Urban Community, Bangladesh.

Bangladesh is a cholera endemic country with a population at high risk of cholera. Toxigenic and non-toxigenic Vibrio cholerae (V. cholerae) can cause cholera and cholera-like diarrheal illness and outbreaks. Drinking water is one of the primary routes of cholera transmission in Bangladesh. The aim of this study was to conduct a comparative assessment of the presence of V. cholerae between point-of-drinking water and source water, and to investigate the variability of virulence profile using molecular methods of a densely populated low-income settlement of Dhaka, Bangladesh. Water samples were collected and tested for V. cholerae from "point-of-drinking" and "source" in 477 study households in routine visits at 6 week intervals over a period of 14 months. We studied the virulence profiles of V. cholerae positive water samples using 22 different virulence gene markers present in toxigenic O1/O139 and non-O1/O139 V. cholerae using polymerase chain reaction (PCR). A total of 1,463 water samples were collected, with 1,082 samples from point-of-drinking water in 388 households and 381 samples from 66 water sources. V. cholerae was detected in 10% of point-of-drinking water samples and in 9% of source water samples. Twenty-three percent of households and 38% of the sources were positive for V. cholerae in at least one visit. Samples collected from point-of-drinking and linked sources in a 7 day interval showed significantly higher odds (P < 0.05) of V. cholerae presence in point-of-drinking compared to source [OR = 17.24 (95% CI = 7.14-42.89)] water. Based on the 7 day interval data, 53% (17/32) of source water samples were negative for V. cholerae while linked point-of-drinking water samples were positive. There were significantly higher odds (p < 0.05) of the presence of V. cholerae O1 [OR = 9.13 (95% CI = 2.85-29.26)] and V. cholerae O139 [OR = 4.73 (95% CI = 1.19-18.79)] in source water samples than in point-of-drinking water samples. Contamination of water at the point-of-drinking is less likely to depend on the contamination at the water source. Hygiene education interventions and programs should focus and emphasize on water at the point-of-drinking, including repeated cleaning of drinking vessels, which is of paramount importance in preventing cholera.

Transmission and Toxigenic Potential of Vibrio cholerae in Hilsha Fish (Tenualosa ilisha) for Human Consumption in Bangladesh.

Fish have been considered natural reservoirs of Vibrio cholerae, the deadly diarrheal pathogen. However, little is known about the role of fish in the transmission of V. cholerae from the Bay of Bengal to the households of rural and urban Bangladesh. This study analyzes the incidence and pathogenic potential of V. cholerae in Hilsha (Tenualosa ilisha), a commonly caught and consumed fish that exhibits a life cycle in both freshwater and marine environments in Bangladesh. During the period from October 2014 to October 2015, samples from the gills, recta, intestines, and scale swabs of a total of 48 fish were analyzed. The fish were collected both at local markets in the capital city Dhaka and directly from fishermen at the river. PCR analysis by targeting V. cholerae species-specific ompW gene revealed that 39 of 48 (81%) fish were positive in at least one of the sample types. Real-time PCR analysis demonstrated that the cholera-causing ctxA gene was detected in 20% (8 of 39) of V. cholerae-positive fish. A total of 158 V. cholerae isolates were obtained which were categorized into 35 genotypic groups. Altogether, 25 O1 and 133 non-O1/O139 strains were isolated, which were negative for the cholera toxin gene. Other pathogenic genes such as stn/sto, hlyA, chxA, SXT, rtxC, and HA-P were detected. The type three secretion system gene cluster (TTSS) was present in 18% (24 of 133) of non-O1/O139 isolates. The antibiotic susceptibility test revealed that the isolates conferred high resistance to sulfamethoxazole-trimethoprim and kanamycin. Both O1 and non-O1/O139 strains were able to accumulate fluid in rabbit ileal loops and caused distinctive cell death in HeLa cell. Multilocus sequence typing (MLST) showed clonal diversity among fish isolates with pandemic clones. Our data suggest a high prevalence of V. cholerae in Hilsha fish, which indicates that this fish could serve as a potential vehicle for V. cholerae transmission. Moreover, the indigenous V. cholerae strains isolated from Hilsha fish possess considerable virulence potential despite being quite diverse from current epidemic strains. This represents the first study of the population structure of V. cholerae associated with fish in Bangladesh.

Can Escherichia coli fly? The role of flies as transmitters of E. coli to food in an urban slum in Bangladesh.

OBJECTIVE: To investigate the transmission of faecal bacteria by flies to food under natural settings. METHODS: Over a period of 2 months, paired (exposed and non-exposed) containers with cooked rice were placed on the ground in kitchen areas in an urban slum area in Dhaka, Bangladesh, and the numbers of flies landing on the exposed rice were counted. Following exposure, the surface of the rice was microbiologically and molecularly analysed for the presence of Escherichia coli and genes of diarrhoeagenic E. coli and Shigella strains. RESULTS: Rice was at greater risk (P < 0·001) of being contaminated with E. coli if flies landed on the rice than if no flies landed on the rice (odds ratio 5·4 (P < 0·001, 95% CI: 2·5-11·7). Mean contamination in exposed rice samples (n = 60) was 3·1 × 103 CFU/g (95% CI: 2·2 × 103-4·0 × 103). Furthermore, for approximately half of the observed fly landings, the average CFU per fly landing was >0·6 × 103 CFU. Genes of diarrhoeagenic E. coli and Shigella species were detected in 39 of 60 (65%) of exposed rice samples. Two fly species were identified: the common housefly (Musca domestica) and the oriental latrine fly (Chrysomya megacephala). CONCLUSION: Flies may transmit large quantities of E. coli to food under field settings. The findings highlight the importance of implementing control measures to minimise exposure of food to flies to ensure food safety. Fly control measures should be considered for the prevention of diarrhoeal diseases caused by E. coli.

Development and Validation of a Novel Real-time Assay for the Detection and Quantification of Vibrio cholerae.

Vibrio cholerae O1 and O139 has been known for its ability to cause epidemics. These strains produce cholera toxin which is the main cause of secretory diarrhea. V. cholerae non-O1 and non-O139 strains are also capable of causing gastroenteritis as well as septicemia and peritonitis. It has been proven that virulence factors such as T6SS, hapA, rtxA, and hlyA are present in almost all V. cholerae strains. It is imperative that viable but non-culturable cells of V. cholerae are also detected since they are also known to cause diarrhea. Thus, the aim of this study was to develop an assay that detects all V. cholerae regardless of their serotype, culturable state, and virulence genes present, by targeting the species specific conserved ompW sequence. The developed assay meets these goals with 100% specificity and is capable of detecting as low as 5.46 copy number of V. cholerae. Detection is rapid since neither lengthy incubation period nor electrophoresis is required. The assay had excellent repeatability (CV%: 0.24-1.32) and remarkable reproducibility (CV%: 1.08-3.7). Amplification efficiencies in the 89-100% range were observed. The assay is more economical than Taqman-based multiplex real-time PCR assays. Compared to other real-time assays, the ompW assay is specific and sensitive, has better repeatability and reproducibility, and is more economical.

Analysis of diarrheagenic potential of uropathogenic Escherichia coli isolates in Dhaka, Bangladesh.

INTRODUCTION: Urinary tract infection is the most frequently diagnosed kidney and urologic disease. METHODOLOGY: Whether the Escherichia coli strains responsible for urinary tract infection (UPEC) carry virulence properties of diarrheagenic E. coli (DEC), 56 UPEC strains were examined for the presence of DEC and UPEC characteristics (e.g. biofilm formation, hemolysis activity, virulence genes). RESULTS: Among 56 UPEC strains, 21 showed capable of biofilm formation and only 5 showed hemolysis activity on sheep blood agar. In Multiplex PCR on assessment of virulence genes related to uropathogenesis; 42% was found positive for papC gene, 27% was fim1 positive, 11% was afa positive and none was found positive for sfa. Most of the isolates were found carrying none of eight diarrhea associated genes (e.g. estA, eltB, vt1, vt2, eaeA, ea, ial and bfpA) as expected. Only seven isolates were found to harbor these genes: five genes i.e., vt2, ial, eltB, bfpA and ea were found in five different isolates and two isolates were positive for estA, among these two, one was found positive for fim1, papC along with estA, a UPEC strain containing virulent gene of ETEC strain. One isolate was found carrying fim1 and vt2 showing the property of EHEC and another isolate was found positive for fim1 and ial, the characteristic of EIEC. One isolate harboring bfpA gene characterized as EPEC and the another one was found to harbor ea gene, characterized as EAEC. CONCLUSIONS: This study observed that most UPEC strains are unique to uropathogenesis, still very few may carry the diarrheagenic property.