Investigating Antimicrobial Resistance and ESBL Producing Gene in Klebsiella Isolates among Neonates and Adolescents in Southern Bangladesh.

Background: Multidrug-resistant (MDR) clones of Klebsiella pneumoniae (Kpn) have been increasingly documented in community-acquired and nosocomial infections all around the globe. Extended-spectrum ß-lactamases (ESBLs) are a rapidly evolving group of ß-lactamase enzymes derived from SHV genes by mutations. This research work aimed to investigate and analyze the widespread prevalence of Kpn antibiotic resistance in different areas of the southern part of Bangladesh. Methods: This particular study was executed and implemented by using 501 clinical samples or isolates from two different hospitals in Chattogram. The disk diffusion method was used to detect Kpn's sensitivity to 16 antibiotics in a drug susceptibility test. By using the PCR technique, the widespread prevalence of antibiotic-resistant gene blaSHV-11 was studied. Sequencing along with phylogenetic analysis was utilized to verify isolates with the blaSHV-11 gene. Results: Almost all of the Kpn isolates were spotted to be antibiotic-resistant. These Kpn isolates were resistant to ß-lactams, aminoglycosides, and quinolones at high levels. The spatial analysis displayed that infections involving Kpn were more common in the urban areas (70%) than in the rural areas (30%). Neonates had substantially higher levels (p < 0.001) of resistance to multidrug than other age groups. Cefepime was identified as the most frequent antibiotic-resistant to all age groups (56.68%). The highest numbers of resistant isolates (36.92%) were found in urine samples. The ESBL gene blaSHV-11 was found in 38% isolates. Conclusion: The significant frequency of MDR Kpn harboring ß-lactamases and AMR genes strongly suggests the requirement to develop effective antimicrobial resistance control and prevention measures in Bangladesh.

Prevalence and molecular characterization of antibiotic resistance and associated genes in Klebsiella pneumoniae isolates: A clinical observational study in different hospitals in Chattogram, Bangladesh.

OBJECTIVE: This study was performed to investigate the prevalence of multidrug resistance and molecular characterization of Klebsiella pneumoniae (KPN) from clinical isolates in the southern region of Bangladesh. Additional analysis of the prevalence of blaNDM-1, blaSHV-11, uge genes of KPN was also carried out among these clinical isolates. METHOD: The study was carried out using 1000 clinical isolates collected from two different hospitals of Chattogram. A drug susceptibility test was performed by the disk diffusion method to detect KPN's response to 16 antibiotics. The presence of antibiotic-resistant and (or) virulent genes blaNDM-1, blaSHV-11, uge were investigated using the PCR technique. Isolates having blaNDM-1, blaSHV-11, uge gene were further validated by sequencing followed by phylogenetic analysis. Phylogenetic relationships among these isolates were determined by Clustal omega and MEGA7. RESULT: A total of 79%, 77%, 74.9%, 71%, 66% and 65% isolates exhibited resistance against cefuroxime, cefixime, cefotaxime, ceftazidime, cefepime and ceftriaxone respectively. The frequency of resistance to other antibiotics varied from 26.5% to 61.8%. PCR analysis showed that 64% of strains harbored blaNDM-1 gene, and 38% strains harbored blaSHV-11 gene. Moreover, 47% of samples were carrying uge gene, and 19% of samples carried blaNDM-1, blaSHV-11, uge genes together. CONCLUSION: In this study, we've analysed the pattern of expression as well as prevalence of blaNDM-1, blaSHV-11, and uge genes in Klebsiella isolates. Upon molecular and statistical analysis, we found a high prevalence of multi-drug resistance KPN strains in the isolates. The Klebsiella isolates were confirmed to harbor multiple ESBL genes and 64% of the isolates were found to be producing NDM-1. As multidrug resistance is an alarming issue, continuous surveillance and routine clinical detection of resistant bacteria and plasmids are necessary to prevent catastrophic public health incidents.

Slow-release nitrogen fertilizers enhance growth, yield, NUE in wheat crop and reduce nitrogen losses under an arid environment.

Higher demands of food led to higher nitrogen application to promote cropping intensification and produce more which may have negative effects on the environment and lead to pollution. While sustainable wheat production is under threat due to low soil fertility and organic matter due to nutrient degradation at high temperatures in the region. The current research explores the effects of different types of coated urea fertilizers and their rates on wheat crop under arid climatic conditions of Pakistan. Enhancing nitrogen use efficiency by using eco-friendly coated urea products could benefit growers and reduce environmental negative effects. A trial treatment included N rates (130, 117, 104, and 94 kg ha-1) and coated urea sources (neem coated, sulfur coated, bioactive sulfur coated) applied with equal quantity following split application method at sowing, 20 and 60 days after sowing (DAS). The research was arranged in a split-plot design with randomized complete block design had three replicates. Data revealed that bioactive sulfur coated urea with the application of 130 kg N ha-1 increased chlorophyll contents 55.0 (unit value), net leaf photosynthetic rate (12.51 µmol CO2 m-2 s-1), and leaf area index (5.67) significantly. Furthermore, research elucidates that bioactive sulfur urea with the same N increased partial factor productivity (43.85 Kg grain Kg-1 N supplied), nitrogen harvest index (NHI) 64.70%, and partial nutrient balance (1.41 Kg grain N content Kg-1 N supplied). The neem-coated and sulfur-coated fertilizers also showed better results than monotypic urea. The wheat growth and phenology significantly improved by using coated fertilizers. The crop reached maturity earlier with the application of bioactive sulfur-coated urea than others. Maximum total dry matter 14402 (kg ha-1) recorded with 130 kg N ha-1application. Higher 1000-grain weight (33.66 g), more number of grains per spike (53.67), grain yield (4457 kg ha-1), and harvest index (34.29%) were obtained with optimum N application 130 kg ha-1 (recommended). There is a significant correlation observed for growth, yield, and physiological parameters with N in the soil while nitrogen-related indices are also positively correlated. The major problem of groundwater contamination with nitrate leaching is also reduced by using coated fertilizers. Minimum nitrate concentration (7.37 and 8.77 kg ha-1) was observed with the application of bioactive sulfur-coated and sulfur-coated urea with lower N (94 kg ha-1), respectively. The bioactive sulfur-coated urea with the application of 130 kg N ha-1 showed maximum phosphorus 5.45 mg kg-1 and potassium 100.67 mg kg-1 in the soil. Maximum nitrogen uptake (88.20 kg ha-1) is showed by bioactive sulfur coated urea with 130 kg N ha-1 application. The total available NPK concentrations in soil showed a significant correlation with physiological attributes; grain yield; harvest index; and nitrogen use efficiency components, i.e., partial factor productivity, partial nutrient balance, and nitrogen harvest index. This research reveals that coating urea with secondary nutrients, neem oil, and microbes are highly effective techniques for enhancing fertilizer use efficiency and wheat production in calcareous soils and reduced N losses under arid environments.

Role of salicylic acid in regulating ethylene and physiological characteristics for alleviating salinity stress on germination, growth and yield of sweet pepper.

BACKGROUND: During a preliminary study, effects of 0, 20, 40, and 60 mM NaCl salinity were assessed on germination rate in relation to electrolyte leakage (EL) in sweet pepper. Results explored significant rises in ethylene evolution from seeds having more EL. It was, therefore, hypothesized that excessive ethylene biosynthesis in plants due to salinity stress might be a root cause of low crop productivity. As salicylic acid is one of the potent ethylene inhibitors, thus SA was used to combat effects of ethylene produced under salinity stress of 60 mM NaCl on different physiological and morphological characteristics of sweet pepper. METHODOLOGY: The effect of 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 mM SA was evaluated on seed germination, growth and yield of sweet pepper cv. Yolo wonder at salinity stress on 60 mM NaCl. Seeds were primed with SA concentrations and incubated till 312 h in an incubator to study germination. Same SA concentrations were sprayed on foliage of plants grown in saline soil (60 mM NaCl). RESULTS: Seeds primed by 0.2 to 0.3 mM SA improved germination rate by 33% due to suppression of ethylene from 3.19 (control) to 2.23-2.70 mg plate-1. Electrolyte leakage reduced to 20.8-21.3% in seeds treated by 0.2-0.3 mM SA compared to 39.9% in untreated seeds. Results also explored that seed priming by 0.3 mM improved TSS, SOD and chlorophyll contents from 13.7 to 15.0 mg g-1 FW, 4.64 to 5.38 activity h-1 100 mg-1 and 89 to 102 ug g-1 compared to untreated seeds, respectively. Results also explore that SA up to 0.2 mM SA applied on plant foliage improved LAI (5-13%), photosynthesis (4-27%), WUE (11-57%), dry weight (5-20%), SOD activity (4-20%) and finally fruit yield (4-20%) compared to untreated plants by ameliorating effect of 60 mM NaCl. Foliar application of SA also caused significant increase in nutrient use efficiency due to significant variations in POD and SOD activities. CONCLUSION: Salicylic acid suppressed ethylene evolution from germinating seeds up to 30% under stress of 60 mM NaCl due to elevated levels of TSS and SOD activity. Foliar application of SA upgraded SOD by lowering POD activity to improve NUE particularly K use efficiency at salinity stress of 60 mM NaCl. Application of 0.2 and 0.3 mM SA emerged as the most effective concentrations of SA for mitigating 60 mM NaCl stress on different physiological and morphological characteristics of sweet pepper.

Presentation and management outcome of tibial infected non-union with Ilizarov technique.

OBJECTIVE: To document the presentation of tibial infected non-union and analyze the management outcome with Ilizarov technique in terms of bone results, functional outcome, bone transport time, external fixation time, external fixation index and any complications. METHODS: This case series was conducted at the Departments of Orthopedic Surgery, National Institute of Rehabilitation Medicine (NIRM), Islamabad and Civil hospital, Quetta over a period of 3-years. RESULTS: There were 56 patients with 53(94.64%) males and 3(5.35%) females. The age range was 16-50 years with a mean of 32.58±9.98years. According to ASAMI criteria, bone results were excellent in 37(66%), good in 10(17.85%), fair in 6(10.71%) and poor in 3(5.35%). The functional results were excellent in 37(66%), good in 9(16%), fair in 7(12.5%) and poor in 3(5.35%). The bone union rate was 98.21%. CONCLUSION: Ilizarov method beautifully addresses the formidable issue of infected non union of tibia with good outcome in terms of bone healing and infection eradication. The treatment period is relatively lengthy and hence patience on part of patient as well as the surgical team is imperative for achieving favourable outcomes.

Safety and efficacy of alternative antibiotic regimens compared with 7 day injectable procaine benzylpenicillin and gentamicin for outpatient treatment of neonates and young infants with clinical signs of severe infection when referral is not possible: a randomised, open-label, equivalence trial.

BACKGROUND: Severe infections remain one of the main causes of neonatal deaths worldwide. Possible severe infection is diagnosed in young infants (aged 0-59 days) according to the presence of one or more clinical signs. The recommended treatment is hospital admission with 7-10 days of injectable antibiotic therapy. In low-income and middle-income countries, barriers to hospital care lead to delayed, inadequate, or no treatment for many young infants. We aimed to identify effective alternative antibiotic regimens to expand treatment options for situations where hospital admission is not possible. METHODS: We did this randomised, open-label, equivalence trial in four urban hospitals and one rural field site in Bangladesh to determine whether two alternative antibiotic regimens with reduced numbers of injectable antibiotics combined with oral antibiotics had similar efficacy and safety to the standard regimen, which was also used as outpatient treatment. We randomly assigned infants who showed at least one clinical sign of severe, but not critical, infection (except fast breathing alone), whose parents refused hospital admission, to one of the three treatment regimens. We stratified randomisation by study site and age (<7 days or 7-59 days) using computer-generated randomisation sequences. The standard treatment was intramuscular procaine benzylpenicillin and gentamicin once per day for 7 days (group A). The alternative regimens were intramuscular gentamicin once per day and oral amoxicillin twice per day for 7 days (group B) or intramuscular procaine benzylpenicillin and gentamicin once per day for 2 days, then oral amoxicillin twice per day for 5 days (group C). The primary outcome was treatment failure within 7 days after enrolment. Assessors of treatment failure were masked to treatment allocation. Primary analysis was per protocol. We used a prespecified similarity margin of 5% to assess equivalence between regimens. This study is registered with ClinicalTrials.gov, number NCT00844337. FINDINGS: Between July 1, 2009, and June 30, 2013, we recruited 2490 young infants into the trial. We assigned 830 infants to group A, 831 infants to group B, and 829 infants to group C. 2367 (95%) infants fulfilled per-protocol criteria. 78 (10%) of 795 per-protocol infants had treatment failure in group A compared with 65 (8%) of 782 infants in group B (risk difference -1.5%, 95% CI -4.3 to 1.3) and 64 (8%) of 790 infants in group C (-1.7%, -4.5 to 1.1). In group A, 14 (2%) infants died before day 15, compared with 12 (2%) infants in group B and 12 (2%) infants in group C. Non-fatal relapse rates were similar in all three groups (12 [2%] infants in group A vs 13 [2%] infants in group B and 10 [1%] infants in group C). INTERPRETATION: Our results suggest that the two alternative antibiotic regimens for outpatient treatment of clinical signs of severe infection in young infants whose parents refused hospital admission are as efficacious as the standard regimen. This finding could increase treatment options in resource-poor settings when referral care is not available or acceptable.

Safety and efficacy of simplified antibiotic regimens for outpatient treatment of serious infection in neonates and young infants 0-59 days of age in Bangladesh: design of a randomized controlled trial.

BACKGROUND: Because access to care is limited in settings with high mortality, exclusive reliance on the current recommendation of 7-10 days of parenteral antibiotic treatment is a barrier to provision of adequate treatment of newborn infections. METHODS: We are conducting a trial to determine if simplified antibiotic regimens with fewer injections are as efficacious as the standard course of parenteral antibiotics for empiric treatment of young infants with clinical signs suggestive of severe infection in 4 urban hospitals and in a rural surveillance site in Bangladesh. The reference regimen of intramuscular procaine-benzyl penicillin and gentamicin given once daily for 7 days is being compared with (1) intramuscular gentamicin once daily and oral amoxicillin twice daily for 7 days and (2) intramuscular penicillin and gentamicin once daily for 2 days followed by oral amoxicillin twice daily for additional 5 days. All regimens are provided in the infant's home. The primary outcome is treatment failure (death or lack of clinical improvement) within 7 days of enrolment. The sample size is 750 evaluable infants enrolled per treatment group, and results will be reported at the end of 2013. DISCUSSION: The trial builds upon previous studies of community case management of clinical severe infections in young infants conducted by our research team in Bangladesh. The approach although effective was not widely accepted in part because of feasibility concerns about the large number of injections. The proposed research that includes fewer doses of parenteral antibiotics if shown efficacious will address this concern.