The Antimicrobial Resistance of Enterotoxigenic Escherichia coli from Diarrheal Patients and the Environment in Two Geographically Distinct Rural Areas in Bangladesh over the Years.

Antimicrobial resistance (AMR) is an unprecedented global health challenge, involving the transfer of bacteria and genes between humans and the environment. We simultaneously and longitudinally determined the AMR of enterotoxigenic Escherichia coli (ETEC) strains isolated from diarrheal patients and an aquatic environment over two years from two geographically distinct locations, Coastal Mathbaria and Northern Chhatak in Bangladesh. A total of 60% and 72% of ETEC strains from the patients in Mathbaria and Chhatak, respectively, were multi-drug resistant (MDR) with a high proportion of ETEC resistant to nalidixic acid (80.7%), macrolides (49.1-89.7%), ampicillin (57.9-69%), and trimethoprim/sulfamethoxazole (55.2%). From the surface water, 68.8% and 30% of ETEC were MDR in Mathbaria and Chhatak, respectively, with a high proportion of ETEC strains resistant to macrolides (87.5-100%), ampicillin (50-75%), ceftriaxone (62.5%), and nalidixic acid (40%). Notably, 80-100% of the ETEC strains were susceptible to tetracycline and quinolones (ciprofloxacin and norfloxacin), both in clinical and aquatic ETEC. The AMR varied by the ETEC toxin types. The patterns of excessive or limited consumption of drugs to treat diarrhea over time in Bangladesh were reflected in the ETEC AMR from the patients and the environment. The high prevalence of MDR-ETEC strains in humans and the environment is of concern, which calls for vaccines and other preventative measures against ETEC.

Epidemiology of Enterotoxigenic Escherichia coli among Children and Adults Seeking Care at Hospitals in Two Geographically Distinct Rural Areas in Bangladesh.

Enterotoxigenic Escherichia coli (ETEC) infections undeniably continue to have substantial morbidity and mortality in younger children; however, limited data are available on the disease burden of older children and adults and on ETEC epidemiology by geographical location at the subnational level. Facility-based surveillance over the years was established to identify patients with ETEC diarrhea in two geographically distinct areas in rural Bangladesh, Chhatak in the north and Mathbaria in the southern coastal area. ETEC was highly prevalent in both areas, while the proportions, toxin types and colonization factors varied by location, season and age groups. Children < 5 years old and adults between 20 and 60 years old were at the highest risk of ETEC diarrhea which required urgent care. This study underscores the importance of capturing subnational and seasonal variations in ETEC epidemiology. ETEC vaccine developers and public health stakeholders may need to target adults between 20 and 60 years of age in addition to young children as new vaccines currently under development become licensed and introduction begins.

Biofilms Comprise a Component of the Annual Cycle of Vibrio cholerae in the Bay of Bengal Estuary.

Vibrio cholerae, an estuarine bacterium, is the causative agent of cholera, a severe diarrheal disease that demonstrates seasonal incidence in Bangladesh. In an extensive study of V. cholerae occurrence in a natural aquatic environment, water and plankton samples were collected biweekly between December 2005 and November 2006 from Mathbaria, an estuarine village of Bangladesh near the mangrove forests of the Sundarbans. Toxigenic V. cholerae exhibited two seasonal growth peaks, one in spring (March to May) and another in autumn (September to November), corresponding to the two annual seasonal outbreaks of cholera in this region. The total numbers of bacteria determined by heterotrophic plate count (HPC), representing culturable bacteria, accounted for 1% to 2.7% of the total numbers obtained using acridine orange direct counting (AODC). The highest bacterial culture counts, including toxigenic V. cholerae, were recorded in the spring. The direct fluorescent antibody (DFA) assay was used to detect V. cholerae O1 cells throughout the year, as free-living cells, within clusters, or in association with plankton. V. cholerae O1 varied significantly in morphology, appearing as distinctly rod-shaped cells in the spring months, while small coccoid cells within thick clusters of biofilm were observed during interepidemic periods of the year, notably during the winter months. Toxigenic V. cholerae O1 was culturable in natural water during the spring when the temperature rose sharply. The results of this study confirmed biofilms to be a means of persistence for bacteria and an integral component of the annual life cycle of toxigenic V. cholerae in the estuarine environment of Bangladesh.IMPORTANCEVibrio cholerae, the causative agent of cholera, is autochthonous in the estuarine aquatic environment. This study describes morphological changes in naturally occurring V. cholerae O1 in the estuarine environment of Mathbaria, where the bacterium is culturable when the water temperature rises and is observable predominantly as distinct rods and dividing cells. In the spring and fall, these morphological changes coincide with the two seasonal peaks of endemic cholera in Bangladesh. V. cholerae O1 cells are predominantly coccoid within biofilms but are rod shaped as free-living cells and when attached to plankton or to particulate matter in interepidemic periods of the year. It is concluded that biofilms represent a stage of the annual life cycle of V. cholerae O1, the causative agent of cholera in Bangladesh.

Acute otitis media during the first two years of life in a rural community in Bangladesh: a prospective cohort study.

The study investigated the burden of acute otitis media (AOM) during the first two years of life in a cohort of 252 newborns in rural Bangladesh using data collected on occurrences of AOM. Trained community health workers (CHWs) conducted household surveillance and picked up cases of AOM using the study algorithm. The incidence rate was 0.9 episodes per child-year observed. Forty-six percent (n=115) of the 252 subjects developed AOM: 36% (n=91) during the first year of life and 10% (n=24) during the second year of life (p<0.001). The age-specific incidence rates of AOM varied; peaks occurred in the 6-12-month age-group and the lowest in the first three months of life. In total, 20% (n=49) of the study subjects had single, 26% (n=66) recurrent, and 54% (n=137) no episode of AOM. Perforation with discharge developed in 85% (n=322) of 375 episodes. The duration of discharge from the ears was < or =6 weeks in 95% of the episodes, but in 5% of the episodes, discharge from the ears continued for >6 weeks. The incidence of AOM was higher in the monsoon season compared to the summer season (p<0.003). The study documented AOM as an important cause of morbidity among rural children up to two years of age in Bangladesh and should be addressed with strategies to overcome the burden of disease.

BUILDING A BETTER GLUTEAL BRIDGE: ELECTROMYOGRAPHIC ANALYSIS OF HIP MUSCLE ACTIVITY DURING MODIFIED SINGLE-LEG BRIDGES.

BACKGROUND: Gluteal strength plays a role in injury prevention, normal gait patterns, eliminating pain, and enhancing athletic performance. Research shows high gluteal muscle activity during a single-leg bridge compared to other gluteal strengthening exercises; however, prior studies have primarily measured muscle activity with the active lower extremity starting in 90 ° of knee flexion with an extended contralateral knee. This standard position has caused reports of hamstring cramping, which may impede optimal gluteal strengthening. HYPOTHESIS/PURPOSE: The purpose of this study was to determine which modified position for the single-leg bridge is best for preferentially activating the gluteus maximus and medius. STUDY DESIGN: Cross-Sectional. METHODS: Twenty-eight healthy males and females aged 18-30 years were tested in five different, randomized single-leg bridge positions. Electromyography (EMG) electrodes were placed on subjects' gluteus maximus, gluteus medius, rectus femoris, and biceps femoris of their bridge leg (i.e., dominant or kicking leg), as well as the rectus femoris of their contralateral leg. Subjects performed a maximal voluntary isometric contraction (MVIC) for each tested muscle prior to performing five different bridge positions in randomized order. All bridge EMG data were normalized to the corresponding muscle MVIC data. RESULTS: A modified bridge position with the knee of the bridge leg flexed to 135 ° versus the traditional 90 ° of knee flexion demonstrated preferential activation of the gluteus maximus and gluteus medius compared to the traditional single-leg bridge. Hamstring activation significantly decreased (p < 0.05) when the dominant knee was flexed to 135 ° (23.49% MVIC) versus the traditional 90 ° (75.34% MVIC), while gluteal activation remained similarly high (51.01% and 57.81% MVIC in the traditional position, versus 47.35% and 57.23% MVIC in the modified position for the gluteus maximus and medius, respectively). CONCLUSION: Modifying the traditional single-leg bridge by flexing the active knee to 135 ° instead of 90 ° minimizes hamstring activity while maintaining high levels of gluteal activation, effectively building a bridge better suited for preferential gluteal activation. LEVEL OF EVIDENCE: 3.

Viral etiology of pneumonia in a cohort of newborns till 24 months of age in Rural Mirzapur, Bangladesh.

Limited information is available on the etiology of acute lower respiratory infection (ALRI) particularly pneumonia in the rural community of developing countries since most etiological studies are carried out in the hospital settings. This study examined the etiology of pneumonia among young children in a rural community of Bangladesh. A cohort of 252 newborns was followed till 24 months of age during 1993-1996. Community health workers (CHWs) identified cases of ALRI during household surveillance and recommended hospitalization. On admission, nasopharyngeal aspirates (NPA) and blood were collected for bacterial and viral identification, and chest x-rays were done. Multiple regression analysis identified factors associated with a viral etiology. Physicians diagnosed 67 pneumonia; 45% of NPA were positive for viral agents of pneumonia, and respiratory syncytial virus (RSV) was predominant (81%); 6 of 48 blood cultures were positive. X-ray was done for 58 cases; 52% had pneumonic consolidation. Of the RSV cases, 33% were found in children without pneumonic consolidation. Children living in a one-room house were 3 times more likely to develop viral pneumonia (odds ratio (OR) = 3.67, CI 1.05-12.83) than children living in a larger house. Counseling on avoiding crowding where a newborn is accommodated might reduce pneumonia incidence.

Organization of the CTX prophage in environmental isolates of Vibrio mimicus.

The organization of the CTX prophage in environmental strains of Vibrio mimicus was investigated. Sixteen hundred non-sucrose fermenting vibrios were examined for ctx gene by hybridization. Out of 1,600 isolates, 6 V. mimicus isolates contained ctxA gene. The organization of CTX prophage was determined by RFLP using ctxA probe. The CTX prophage integrated at a single site in V. mimicus genome which was present as a single copy flanked by at least a single RS element. Ribotype pattern revealed that a particular clone of V. mimicus acquired the CTXPhi in the aquatic environment. This study demonstrated that V. mimicus could act as a reservoir of CTXPhi in the aquatic environment.

Virulence properties of rough and smooth strains of Vibrio cholerae O1.

A comparative study was carried out to see the differences in pathogenicity of rough and smooth strains. A total of 10 strains including 5 each of rough and smooth strains of Vibrio cholerae O1 were tested and found positive for toxin production by enzyme-linked immunosorbent assay (ELISA) in Richardson's and AKI media. All the smooth and rough strains, except one, showed a titre of 1: 10 and 1: 100 in Richardson's and AKI media, respectively. Both types of strains produced enterotoxin in rabbit ileal loop (RIL). The differences in multiplication abilities of smooth and rough strains in RIL were statistically significant (P <0.05). However, these differences in multiplying abilities did not influence the adherence potential or enterotoxin production as there was no significant difference (P >0.05) between these properties. This study demonstrated that the rough strains are equally pathogenic and as important as smooth strains.

Variation of toxigenic Vibrio cholerae O1 in the aquatic environment of Bangladesh and its correlation with the clinical strains.

The diversity of toxigenic V. cholerae O1 in the aquatic environment of Bangladesh is not known. A total of 18 environmental and 18 clinical strains of toxigenic V. cholerae O1 were isolated simultaneously from four different geographical areas and tested for variation by the pulsed-field gel electrophoresis method. Environmental strains showed diversified profiles and one of the profiles was common to some environmental strains and most clinical strains. It appears that one clone has an advantage over others to cause disease. These findings suggest that the study of the molecular ecology of V. cholerae O1 in relation to its environmental reservoir is important in identifying virulent strains that cause disease.