Effect of Endophytic Serratia marcescens Isolated from Bryophyllum pinnatum against Clinical Bacterial Isolates

The world's health is being threatened by antimicrobial resistance (AMR). According to the World Health Organization, it is one of the top ten worldwide public health problems facing humanity. Serratia marcescens is an opportunistic pathogen that can be isolated from soil, plants, water, and air. Additionally, Serratia species offer a valuable supply of secondary metabolites that are comparatively underutilised and may have anti-MDR pathogenic properties. The present research aimed to determine the antibacterial potential of Serratia marcescens isolated from the leaves of Bryophyllum pinnatum against clinical bacterial isolates. The leaves of Bryophyllum pinnatum were collected, surface sterilised, cultured at 37 °C for 24 hours and identified utilising viteks 2 automated techniques and molecular methods. The crude metabolites extract of Serratia marcescens were extracted and utilised for antibacterial susceptibility testing using agar healthy diffusion methods. The data were measured in the diameter zone of inhibition. This study revealed six endophytic bacteria were isolated from Bryophyllum pinnatum following standard microbiological culture methods. The endophytic bacteria isolate tag L03 was found to be Gram-Negative Rod. The isolate was tentatively identified as Serratia ficaria and molecularly identified as Serratia marcescens. The metabolites of Serratia marcescens endophytes revealed a significant antibacterial activity on Klebsiella pneumoniae with a diameter zone of inhibition of 17.7 mm at 100% concentration, followed by Staphylococcus sciuri with a 12.7 mm diameter zone of inhibition. These results suggested that endophytic bacteria Serratia marcescens were isolated from the leaves of Bryophyllum pinnatum and had shown potent antibacterial activity that could be employed to create new antibacterial agents

Neonatal sepsis and mortality in low-income and middle-income countries from a facility-based birth cohort: an international multisite prospective observational study

Background Neonatal sepsis is a primary cause of neonatal mortality and is an urgent global health concern, especially within low-income and middle-income countries (LMICs), where 99% of global neonatal mortality occurs. The aims of this study were to determine the incidence and associations with neonatal sepsis and all-cause mortality in facility-born neonates in LMICs. Methods The Burden of Antibiotic Resistance in Neonates from Developing Societies (BARNARDS) study recruited mothers and their neonates into a prospective observational cohort study across 12 clinical sites from Bangladesh, Ethiopia, India, Pakistan, Nigeria, Rwanda, and South Africa. Data for sepsis-associated factors in the four domains of health care, maternal, birth and neonatal, and living environment were collected for all mothers and neonates enrolled. Primary outcomes were clinically suspected sepsis, laboratory-confirmed sepsis, and all-cause mortality in neonates during the first 60 days of life. Incidence proportion of livebirths for clinically suspected sepsis and laboratory-confirmed sepsis and incidence rate per 1000 neonate-days for all-cause mortality were calculated. Modified Poisson regression was used to investigate factors associated with neonatal sepsis and parametric survival models for factors associated with all-cause mortality. Findings Between Nov 12, 2015 and Feb 1, 2018, 29 483 mothers and 30 557 neonates were enrolled. The incidence of clinically suspected sepsis was 166·0 (95% CI 97·69–234·24) per 1000 livebirths, laboratory-confirmed sepsis was 46·9 (19·04–74·79) per 1000 livebirths, and all-cause mortality was 0·83 (0·37–2·00) per 1000 neonate-days. Maternal hypertension, previous maternal hospitalisation within 12 months, average or higher monthly household income, ward size (>11 beds), ward type (neonatal), living in a rural environment, preterm birth, perinatal asphyxia, and multiple births were associated with an increased risk of clinically suspected sepsis, laboratory-confirmed sepsis, and all-cause mortality. The majority (881 [72·5%] of 1215) of laboratory-confirmed sepsis cases occurred within the first 3 days of life. Interpretation Findings from this study highlight the substantial proportion of neonates who develop neonatal sepsis, and the high mortality rates among neonates with sepsis in LMICs. More efficient and effective identification of neonatal sepsis is needed to target interventions to reduce its incidence and subsequent mortality in LMICs. Funding Bill & Melinda Gates Foundation

Effects of antibiotic resistance, drug target attainment, bacterial pathogenicity and virulence, and antibiotic access and affordability on outcomes in neonatal sepsis: an international microbiology and drug evaluation prospective substudy (BARNARDS)

Background Sepsis is a major contributor to neonatal mortality, particularly in low-income and middle-income countries (LMICs). WHO advocates ampicillin–gentamicin as first-line therapy for the management of neonatal sepsis. In the BARNARDS observational cohort study of neonatal sepsis and antimicrobial resistance in LMICs, common sepsis pathogens were characterised via whole genome sequencing (WGS) and antimicrobial resistance profiles. In this substudy of BARNARDS, we aimed to assess the use and efficacy of empirical antibiotic therapies commonly used in LMICs for neonatal sepsis. Methods In BARNARDS, consenting mother–neonates aged 0–60 days dyads were enrolled on delivery or neonatal presentation with suspected sepsis at 12 BARNARDS clinical sites in Bangladesh, Ethiopia, India, Pakistan, Nigeria, Rwanda, and South Africa. Stillborn babies were excluded from the study. Blood samples were collected from neonates presenting with clinical signs of sepsis, and WGS and minimum inhibitory concentrations for antibiotic treatment were determined for bacterial isolates from culture-confirmed sepsis. Neonatal outcome data were collected following enrolment until 60 days of life. Antibiotic usage and neonatal outcome data were assessed. Survival analyses were adjusted to take into account potential clinical confounding variables related to the birth and pathogen. Additionally, resistance profiles, pharmacokinetic–pharmacodynamic probability of target attainment, and frequency of resistance (ie, resistance defined by in-vitro growth of isolates when challenged by antibiotics) were assessed. Questionnaires on health structures and antibiotic costs evaluated accessibility and affordability. Findings Between Nov 12, 2015, and Feb 1, 2018, 36 285 neonates were enrolled into the main BARNARDS study, of whom 9874 had clinically diagnosed sepsis and 5749 had available antibiotic data. The four most commonly prescribed antibiotic combinations given to 4451 neonates (77·42%) of 5749 were ampicillin–gentamicin, ceftazidime–amikacin, piperacillin–tazobactam–amikacin, and amoxicillin clavulanate–amikacin. This dataset assessed 476 prescriptions for 442 neonates treated with one of these antibiotic combinations with WGS data (all BARNARDS countries were represented in this subset except India). Multiple pathogens were isolated, totalling 457 isolates. Reported mortality was lower for neonates treated with ceftazidime–amikacin than for neonates treated with ampicillin–gentamicin (hazard ratio [adjusted for clinical variables considered potential confounders to outcomes] 0·32, 95% CI 0·14–0·72; p=0·0060). Of 390 Gram-negative isolates, 379 (97·2%) were resistant to ampicillin and 274 (70·3%) were resistant to gentamicin. Susceptibility of Gram-negative isolates to at least one antibiotic in a treatment combination was noted in 111 (28·5%) to ampicillin–gentamicin; 286 (73·3%) to amoxicillin clavulanate–amikacin; 301 (77·2%) to ceftazidime–amikacin; and 312 (80·0%) to piperacillin–tazobactam–amikacin. A probability of target attainment of 80% or more was noted in 26 neonates (33·7% [SD 0·59]) of 78 with ampicillin–gentamicin; 15 (68·0% [3·84]) of 27 with amoxicillin clavulanate–amikacin; 93 (92·7% [0·24]) of 109 with ceftazidime–amikacin; and 70 (85·3% [0·47]) of 76 with piperacillin–tazobactam–amikacin. However, antibiotic and country effects could not be distinguished. Frequency of resistance was recorded most frequently with fosfomycin (in 78 isolates [68·4%] of 114), followed by colistin (55 isolates [57·3%] of 96), and gentamicin (62 isolates [53·0%] of 117). Sites in six of the seven countries (excluding South Africa) stated that the cost of antibiotics would influence treatment of neonatal sepsis

Biofilm forming Enterococci and their Status as Emerging Multidrug Resistant Bacteria

Urine specimens were collected aseptically and inoculated directly onto CLED and MacConkey agar medium and incubated overnight. Single target colonies were sub cultured, and identification was based on culture, morphology, and biochemical characteristics. Enterococci isolates were subjected to biofilm assay (Tube method), and resistance pattern was determined for both biofilm-formers and non-biofilm-formers. Genomic DNA was extracted using the Bioneer kit, and the gene for virulence was detected by PCR and agarose gel electrophoresis. 148 Patients having one or more urinary symptoms were the study population. The aim of the study was to determine the multidrug resistant ability of biofilm-forming Enterococci. A number of growths were encountered; Enterococci accounted for only 9.46% (14), the highest prevalence was seen in the age group 21-30 (35.71%), and females (64.29%) were more prone to enterococcal infection than their males (35.71) counterpart. There is a relationship between biofilm production and antibiotic resistance because multidrug resistant Enterococci isolate produced bands against esp gene with an average of 510kbp. Biofilm forming strains showed the highest resistance to gentamicin and penicillin (83.3%), and vancomycin can be considered a good alternative therapy in enterococcal UTIs because of its lower resistance (75%)