Standard Biochemical Tests Research Articles

Strengthening identification and characterization of causes of perinatal deaths in Kaski district of Nepal (Perinatal MITS Nepal)

BackgroundThree million babies die in the early neonatal period while 2.6 million are stillborn per year worldwide, and one of three deaths can be prevented. The perinatal mortality rate in Nepal is around 31 per 1000 births. Although the perinatal and neonatal death rates have decreased recently, it still poses a major challenge to the health system of Nepal. The objective of the study is to determine the causes of perinatal deaths by integrating Minimally Invasive Tissue Sampling (MITS) in hospital perinatal deaths and incorporating verbal autopsy in community deaths in Kaski district of Nepal.MethodsThe study will be conducted among the perinatal deaths reported in the five hospitals implementing the Maternal and Perinatal Death Surveillance and Response (MPDSR) system in Kaski district of Nepal. We will also conduct verbal autopsy (VA) among community perinatal deaths reported in the district. All the perinatal deaths reported in the study sites will be enrolled in the first stage of the study. Minimally Invasive Tissue Sampling (MITS) will be conducted among the consenting cases of perinatal deaths to retrieve relevant tissue samples and specimens. The specimens will undergo standard histopathological, microbiological, biochemical, and molecular tests. The “Cause of Death Panel” will finalize MITS informed cause of death following the customized protocol for the project and the cause so derived will be compared with that obtained by the review of deaths by the MPDSR committees of the hospitals. The Female Community Health Volunteers will be trained and mobilized to identify community perinatal deaths and trained personnel will conduct VA. Community engagement activities will be conducted to provide awareness to prevent perinatal deaths.DiscussionThe mechanism of counting and accounting for deaths in a systematic manner is important and it can provide evidence to determine changes in clinical practice and to develop guidelines and training packages for preventive measures. The outcome will be helpful to standardize methods to establish the accurate causes of perinatal deaths and develop strategies to minimize the deaths. The selected pathological investigations can be integrated into the existing death surveillance system in order to effectively determine the causes of death.

Prevalence of Extended Spectrum Beta-Lactamase Producing Bacteria in Patients with Wound Infections Attending Tertiary Hospitals in Enugu, Nigeria

Background: The problem of antibiotics usage against bacterial infection is the modifications of such antibiotics by the bacteria thereby rendering them ineffective. Extended spectrum β-lactamase (ESBL) producing bacteria invading wound infections may lead to long term hospitalization, financial burden and limited antibiotics for therapy. The goals of this study were to determine the prevalence of ESBL-producing bacteria colonization of wound infections among individuals with non-healing wounds and the antibiotic susceptibility pattern of the ESBL isolates. Methodology: The study adopted a cross-sectional research design. A total of 266 samples were collected from different wound infections which included diabetic foot ulcers, burn wounds, post-surgical wounds, non-diabetic foot ulcers, pressure ulcers, accident and open cancer wounds that met the inclusion criteria. The patients were consecutively selected. That is, any individual that has a wound and was willing to participate was selected. A structured questionnaire was administered to the patients to obtain information on demographic characteristics, antibiotic usage, and duration of infection, herbal medication, type and site of wound. Identification of bacterial isolates was done using colony/ microscopic morphology, gram stain reaction and standard biochemical tests. Antibiotic susceptibility testing was done using modified Kirby - Bauer disc diffusion method. ESBL detection was done following the recommendations by the Clinical and Laboratory Standard Institute (CLSI) which involves a 2-step approach of initially screening for ESBL producers and phenotypic confirmatory test using a combination disc test method. Molecular screening of the genes encoding for ESBLs was done at the Central Science Laboratory, University of Nigeria, Nsukka. Results: A total of 196 isolates were recovered from the wound swabs. Pseudomonas aeruginosa 56 (28.6%) was the leading organism causing wound infection followed by Staphylococcus aureus 24 (12.2%). The bacteria isolates showed that 157 (80.1%) were gram negatives as against 39 (19.9%) that were gram positive bacteria. Among the gram-negative bacteria isolates, 21.7% (34/157) were confirmed as ESBL producers. The ESBL- producers were Pseudomonas aeruginosa, Esherichia coli, Proteus vulgaris, Proteus mirabilis, Klebsiella oxytoca, Klebsiella pneumonia and Klebsiella granulomatis at frequencies of 41.2%, 20.6%,14.7%,11.8%,5.9%,2.9% and 2.9% respectively. This showed that these isolates have the ability to resist penicillins and cephalosporins of the first, second and third generations. The ESBL-producing bacteria isolated exhibited high degree of multidrug resistance especially to tetracyclines, cefuroxime, ceftriaxone, cefotaxime and ceftazidime. The antibiotics amikacin was sensitive to most of the ESBL-producing bacteria isolated. The assessment of the risk factors showed that none of the variables was statistically significant, though those risk factors were still important in evaluating wound infections. Both hospital and community acquired infections showed no statistical significance (P= 0.072) which means that both had the same degree of pathogenicity. Among the 10 samples screened for ESBLs genes namely bla SHV, bla TEM, bla CTX-M, bla GES, and bla OXA-50, only bla OXA- 50 was detected in 8 out of the ten samples. Thus, the persistent gene circulating in this region is bla OXA-50, which confers high rate of infection and persistence. Conclusion: The presence of ESBL-producing bacteria in wounds remains a challenging issue, as the majority of the patients may suffer from long term infected wounds due to treatment failure.

Extended spectrum betalactamase and Carbapenemase producing gram negative bacteria from healthcare workers gowns at Debre Berhan Comprehensive Specialized Hospital, Ethiopia

Extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing (CP) gram-negative bacteria are the major public health concerns. Gowns used by healthcare workers (HCWs) in daily practice are a source of hospital-acquired infections in hospital settings. The study aimed to determine the prevalence of extended-spectrum beta-lactamase and carbapenemase-producing gram-negative bacteria from gowns of healthcare workers at Debre Berhan Comprehensive Specialized Hospital, Amhara Regional State, Ethiopia. A hospital-based cross-sectional study was conducted from August to October 2022. A total of 226 swab samples were collected from gowns of HCWs and cultured on MacConkey agar. Bacterial identifications were done by standard biochemical tests. Screening for ESBL and carbapenemase production was done using CHROME agar. Confirmation of ESBL and carbapenemase production was done by the combination disk method and modified carbapenem inactivation method, respectively. Data were analyzed by using SPSS version 25. The overall contamination rate of gowns was 46.9% (106/226). Among 226 swab samples, a total of 117 (51.7%) gram-negative bacteria were isolated. Among these, the most frequent isolates were E. coli, accounted 36 (30.8%) followed by K. pneumoniae, 26(22.2%). The overall multi-drug resistance (MDR) rate was 65 (55.6%). Of the 117 isolates, 17(14.5%) and 12(10.3%) were ESBL and carbapenemase producers respectively. Gown type (p = 0.041), laundering practice (p = 0.045), number of gowns (p = 0.002), and gown washing frequency per week (p = 0.017).were significantly associated with bacterial contamination of gowns. In this study, the prevalence of ESBL and CP Gram-negative bacteria from gowns of healthcare workers was found to be alarming. Therefore, strict infection prevention and control practices, as well as good hygienic practices, should be implemented to reduce and prevent cross-contamination and the spread of antimicrobial-resistant strains in hospital settings.

Clinical and molecular analysis of ESBL, carbapenemase, and colistin-resistant bacteria in UTI patients.

Uropathogens, particularly bacteria, can infect any part of the urinary tract and cause bacteriuria. Our study aimed to examine the antibiotic-resistant profile, associated risk factors, and phenotypic and genotypic features of ESBL, carbapenemase, and mcr resistance genes in multidrug-resistant bacteria. Samples were inoculated on culture media, identified using standard biochemical tests, and species confirmation was performed via 16S rRNA gene amplification. Furthermore, antibiotic susceptibilities were evaluated using the Kirby-Bauer disc diffusion method. The phenotypically confirmed resistant strains were further inspected for ESBL, carbapenemases, and mcr variants using PCR. Merely 57.24% (83/145) of the samples exhibited growth. Of these, 39.70% (33/83) were identified as Klebsiella pneumoniae, 27.70% (23/83) as Escherichia coli, 10.80% (9/83) as Pseudomonas aeruginosa, 9.60% (8/83) as Staphylococcus aureus, 7.20% (6/83) as Proteus mirabilis, and 4.80% (4/83) as Staphylococcus saprophyticus. Overall, 22.54% (16/71) of the gram-negative strains were confirmed molecularly to have resistant genes. The ESBL - producers accounted for 21.74% (5/23) of E. coli, 21.21% (7/33) of K. pneumoniae, and 22.22% (2/9) of P. aeruginosa. Likewise, carbapenemase-harboring strains included 6.06% (2/33) of K. pneumoniae, 4.35% (1/23) of E. coli, and 11.11% (1/9) of P. aeruginosa. Notably, 3.03% (1/33) of K. pneumoniae, 8.70% (2/23) of E. coli, and 11.11% (1/9) of P. aeruginosa strains tested positive for the mcr-1 gene. None of the Proteus strains showed any resistant genes. The most common variants were blaSHV-11 (non-ESBL) and blaCTX-M-15 (ESBL) accounted for 28.57% (4/14) each, blaTEM-116 accounted for 14.29% (2/14), blaSHV-1, blaSHV-75, blaTEM-1 and blaOXA-1 accounted for 7.14% (1/14) each of the ESBL. Similarly, the carbapenemase variants included blaOXA-48, blaNDM-1, blaVIM-1, and blaKPC-2, each accounting for 25.0% (1/4), while 37.50% (6/16) of the strains exhibited co-existence of different gene variants. Based on our findings, it can be concluded that females, especially those in middle age, were more infected. These pathogens exhibited a wide range of ESBL, carbapenemase, and mcr-1 variants. Imipenem was suggested as the preferred medication.

Antibiotic Susceptibility Pattern of Bacterial Isolates from Infected Diabetic Foot Ulcer in Patients of Type 2 Diabetes Mellitus Presenting at Hayatabad Medical Complex Peshawar

Objectives: This study aimed to determine the antibiotic resistance profiles of bacterial isolates obtained from diabetic foot ulcers (DFUs) in type 2 diabetes patients at Hayatabad Medical Complex, Peshawar, by identifying the most prevalent bacterial species and their corresponding resistance patterns. Materials and Methods: This cross-sectional study was conducted over six months from March to September 2024, involving 120 clinically diagnosed patients with infected diabetic foot ulcers. Bacterial isolates were obtained from wound swabs and identified using standard cultural and biochemical tests. Antibiotic susceptibility was determined through the disc diffusion method. Results: The results indicated that 93.3% of wound swabs showed positive bacterial growth, predominantly gram-negative bacteria, with Escherichia coli (28%) and Klebsiella pneumoniae (22%) being the most prevalent isolates. Polymicrobial infections were found in 18% of samples. Resistance rates were notably high for ampicillin (72%) and ciprofloxacin (55%), while carbapenems and piperacillin-tazobactam demonstrated higher sensitivity. Among gram-positive isolates, methicillin-resistant Staphylococcus aureus (MRSA) was detected in 60% of cases. Conclusion: The research underscores the significant prevalence of multidrug-resistant (MDR) bacteria in diabetic foot ulcers, emphasizing the critical need for innovative approaches to antibiotic treatment and robust infection control strategies.

Identification of beta-lactamase genes and molecular genotyping of multidrug-resistant clinical isolates of Klebsiella pneumoniae

BackgroundKlebsiella pneumoniae is a clinically relevant pathogen that has raised considerable public health concerns. This study aims to determine the presence of beta-lactamase genes and perform molecular genotyping of multidrug-resistant (MDR) K. pneumoniae clinical isolates.MethodsClinical isolates of MDR K. pneumoniae were collected from educational hospitals affiliated with Babol University of Medical Sciences. The isolates of K. pneumoniae were identified through standard microbial and biochemical tests. Antibiotic resistance was assessed using disk diffusion, modified Hodge test (MHT), combined disk, and polymerase chain reaction (PCR) methods. Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR was performed for molecular typing.ResultsA total of 42 MDR K. pneumoniae isolates were obtained from various clinical specimens. The highest antibiotic resistance was observed for ampicillin (100%), while the lowest resistance was noted for amikacin (19.04%). The MHT indicated that 38.09% of K. pneumoniae isolates produced carbapenemase enzymes. Metallo-beta-lactamase (MBL) production was found in 54.76% of isolates. Molecular detection of beta-lactamase genes revealed the presence of blaNDM (21.42%), blaKPC (42.85%), blaTEM (76.19%), blaSHV (47.16%), and blaCTX−M (80.95%) genes. ERIC-PCR molecular typing identified seven distinct genetic patterns among the isolates.ConclusionsThis investigation demonstrates the high resistance levels of K. pneumoniae strains. The beta-lactamase genes with the highest and lowest frequencies correspond to blaCTX−M and blaNDM genes, respectively. ERIC-PCR dendrograms suggest a common origin for K. pneumoniae clinical isolates and the propagation of similar clones within hospital wards. These findings indicate that K. pneumoniae isolates are highly virulent, necessitating the development of more effective resistance-fighting techniques and gene transfer research.Clinical trial numberNot applicable.

Detection and Molecular Characterization of Escherichia coli from Wastewater Environments in Two University Campuses in Nigeria.

Escherichia coli (E. coli) is the most prominent bacterial pathogen that causes urinary tract infections (UTIs), and the rate of resistance to most used antibiotics is alarmingly increasing. This study assessed the hostel gutters of two Nigerian universities, the University of Nigeria, Nsukka (UNN) and Kogi State University, Anyigba (KSU), for E. coli and its antimicrobial resistance genes (ARGs). Oxoid Chromogenic UTI agar was used to isolate uropathogenic E. coli (UPEC), identified using standard biochemical tests. The virulence and resistance genes of the isolates were further characterized using molecular techniques. A total of 906 UPEC were isolated in this study, of which 63 isolates were selected for antimicrobial susceptibility tests. The UPEC isolates showed 100% resistance to amoxicillin/clavulanic acid, vancomycin, and penicillin G, while a complete sensitivity of the isolates to meropenem and ciprofloxacin was observed. The index of isolates showing multidrug resistance ranged from 0.33 to 0.73. The level of multiple drug resistance (MDR) exhibited by the UPEC isolates from effluent was significantly higher compared to those from influent (p < 0.05). The ARGs detected were blaOXA-1 8 (38.1%), blaCTX-M3 8 (38.1%), and ant(2)-la 20 (95.2%). Virulence genes encodings beta-glucuronidase (uidA) and hemolysin A (hlyA) were detected in 95.2% of UPEC isolates. The current study showed that UPEC is widely distributed in the environment of two Nigerian universities. The index range of MDR and the circulation of ARGs and virulence genes in the environment suggest a potential health concern, thus warranting further investigation.

Isolation and Molecular Identification of Lipolytic Bacteria from Oil Contaminated Soil

Soil, the upper layer of the Earth's surface, a complex and dynamic ecosystem that supports plant growth, provides water and nutrients, and purifies the air and water. Comprising a mixture of organic matter, minerals, gases, liquids, and organisms, soil is a natural resource that plays a vital role in many ecosystem functions. However, human activities such as land use change, urbanization, deforestation, and intensive agriculture can lead to soil degradation and loss of soil resources. Soil organisms, including bacteria, fungi, and earthworms, contribute to organic matter decomposition, nutrient cycling, and soil structure improvement, but can also compete with plants for nutrients. Soil pollution, caused by industrial activities, agricultural chemicals, and waste disposal, poses a significant threat to human health and the environment. Bioremediation, using microorganisms to break down pollutants, offers a promising solution for soil remediation. This study focuses on the isolation and identification of lipolytic bacteria from oil-contaminated soil, highlighting the importance of these microorganisms in degrading lipids and pollutants.In this study, the soil sample was examined for the presence of lipase-positive strains using a screening method and strains were identified based on the standard physical, biochemical and molecular tests. The 16S rRNA gene sequence analysis used identified the strains as Acinetobacter juniiand Bacillus tropicus.

Diarrhoeagenic Escherichia Coli Isolated from Children with Acute Diarrhoea by Culture and Multiplex PCR in Tertiary Care Hospitals of Dhaka, Bangladesh

Background: Diarrhoeal diseases are a major childhood health problem and an important cause of morbidity and mortality, especially in developing countries. Objective: This study aimed to detect Diarrhoeagenic Escherichia coli (DEC) by detecting virulence genes (lt, st, bfp, and aat) and to determine the antimicrobial susceptibility pattern of all identified DEC isolates in fecal samples. Methodology: This cross-sectional study was carried out in the Department of Microbiology at Dhaka Medical College Hospital, Dhaka, Bangladesh and Dhaka Shishu Hospital, Dhaka, Bangladesh from July, 2012 to June, 2013 for around one year. Identification of Diarrhoeagenic Escherichia coli (DEC) in fecal samples from children under 5 years old was done by culture and standard biochemical tests. Virulence factor genes for DEC were detected by Polymerase Chain Reaction (PCR). Susceptibility to antimicrobial agents of all identified DEC isolates was observed by disc diffusion method. Results: Out of total 200 stool samples, E. coli was isolated in 130(65.0%). The prevalence of DEC was 52(40.0%) and the most common was enterotoxigenic Escherichia coli (55.8%) followed by enteropathogenic Escherichia coli (34.6%) and enteroaggregative Escherichia coli (26.9%). Among 52 DEC, 12(23.1%) contained more than one pathogenic genes of DEC in various combinations. Colistin, imipenem and amikacin were found to be the most effective against isolated DEC. Most of the strains were resistant to ampicillin, erythromycin, tetracycline and nalidixic acid. Conclusion: Differentiation between the Diarrhoeagenic Escherichia coli pathotypes is of great importance since they are involved in acute diarrhoeal diseases and may require specific antimicrobial chemotherapy. Bangladesh Journal of Medical Microbiology, January 2024;18(1):17-21

Profile of Bacteria Isolated from the Cell Phones of Health Care Providers in a Hospital Setting in Cameroon

Health care providers are an integral part of the era of mobile phones. During various activities linked to health care services, they come in contact with their cell phones regularly. These cell phones act as a perfect substrate for nosocomial pathogens, especially in hot–humid conditions, and may serve as a vehicle in transmitting nosocomial infections. This study aimed at determining the profile of bacteria isolated from the cell phones of healthcare providers. A descriptive cross-sectional study was carried out from 1 April to 3 June 2023, where 115 swab samples were collected from the cell phones of health care providers (laboratory personnel, nurses/midwives and doctors) at the Regional Hospital Bamenda. These swabs were inoculated on blood, chocolate and Mac Conkey plates, and the bacteria were identified according to standard microbiological methods and biochemical tests to the genus/specie level. Data were statistically analyzed using the Statistical Package for Social Sciences (SPSS) version 23. The results were presented in frequencies and proportions. The chi square test was used to compare proportions between variables, and the results were considered statistically significant when p &lt; 0.05. The bacteria isolated from these cell phones included coagulase-negative Staphylococci (CoNS), Staphylococcus aureus, Streptococcus species, Pseudomonas aeroginosa, Escherichia coli, Bacillus species and Neisseria species. All the cell phones of the laboratory personnel were contaminated, followed by those of the nurses/midwives (38; 33.0%) and, lastly, by those of the medical doctors (24; 20.9%). No statistically significant difference was observed between the three categories of health care providers with respect to the presence or absence of bacteria on their cell phones. This study presented that the mobile phones of health care providers are a risk of nosocomial pathogens. The result implies that there is an urgent need to implement and emphasize strategies such as hand washing and decontamination of mobile phones to limit nosocomial infections in the hospital.

Extended-spectrum β-lactamase and carbapenemase producing Enterobacteriaceae among patients suspected with surgical site infection at Hospitals in Southern Ethiopia.

Extended-spectrum β-lactamase and carbapenemase-producing Enterobacteriaceae are an increasing problem for patients today. Data on clinical samples for ESBL and carbapenemase-producing Enterobacteriaceae for surgical site infection patients in developing countries are limited, including Ethiopia, mainly due to resource constraints. Hence, this study aimed to determine the prevalence of extended-spectrum beta-lactamase- and carbapenemase-producing Enterobacteriaceae among patients suspected to have surgical site infection at Hospital in Southern Ethiopia. A hospital-based cross-sectional study was conducted on 422 suspected surgical site infections from June 1, 2022 to August 30, 2022 at Hospitals in Southern Ethiopia. Sociodemographic and clinical data were obtained by using a structured questionnaire. Clinical samples (pus, pus aspirates, and wound swabs) were collected aseptically and processed within 30 min by placing the swabs in sterile test tubes containing sterile normal saline (0.5 mL). Samples were cultured on blood and MacConkey agar plates. All positive cultures were characterized by colony morphology, Gram staining, and standard biochemical tests. Antimicrobial sensitivity tests were performed using Kirby Baur disk diffusion on Mueller-Hinton agar. ESBL production was confirmed using a double-disc synergy test (DDST) method. Carbapenemase production was assessed using the modified Hodge test. Logistic regression analysis was used to determine associated factors. A P-value < 0.05 were considered statistically significant. Bacteria belonging to the order Enterobacterales were cultured in 23.7% out of 422 patients with suspected surgical wound infection. Of all the isolates, Enterobacteriaceae (69 isolates) were the most frequent, with E. coli (29/69) followed by K. pneumoniae (14/69). Of 69 Enterobacteriaceae isolates, 66.6 % (46/69) were positive for ESBL production, and 21.7 (15/69) were positive for carbapenemase-producing Enterobacteriaceae. The majority of Enterobacteriaceae isolates showed sensitivity to meropenem (72.1%); however revealed 63.9% and 70.5% were resistant to gentamicin and ciprofloxacin, respectively. Similarly, a higher resistance rate to cefepime (91.8%), amoxicillin-clavulanic acid (98.4%), ceftriaxone (95.1%), and ceftazidime (91.8%). MDR rate of Enterobacteriaceae isolates was 25/61 (41%) among patients suspected for surgical site infection. The Multivariable analysis revealed that length of hospital stay in hospital [AOR = 3.81 (95% CI 2.08-6.95)] remained statistically significant factor associated with surgical site infection due to ESBL producing Enterobacteriaceae. Study results showed the severity of ESBL-producing Enterobacteriaceae is critical and CPE is alarming. Meropenem is the most effective antibiotic against the ESBL-producing Enterobacteriaceae. MDR rate of Enterobacteriaceae isolates was 61 (61%) among patients suspected for surgical site infection. Therefore, antibiotic selection should be based on the results of the culture and sensitivity tests.

Investigating the Prevalence of Bacterial Infections in Patients with Coronavirus Disease 2019 Hospitalized in Intensive Care Unit and Determining their Antibiotic Resistance Patterns.

COVID-19 patient hospitalization, particularly in intensive care units, exposes them to bacterial and fungi co-infections, which can have very serious consequences, including increased mortality. In addition, antibiotic resistance among pathogens is a hidden threat behind COVID-19. In the period from 2020 September to 2021 August, bacterial isolates from COVID-19 patients admitted to the ICU of Sina Hospital in Hamadan, Iran, were collected and identified based on standard biochemical tests. COVID-19 cases were confirmed based on clinical symptoms, computed tomography, and polymerase chain reaction. Antimicrobial susceptibility tests were conducted using disc diffusion and broth microdilution methods. In total, 207 bacterial isolates were collected, with Klebsiella pneumoniae account-ing for 69 (33.33%) and Acinetobacter baumannii accounting for 59 (28.15%). The frequency and percentage of isolated bacteria were as follows: Alcaligenes species 28 (13.59%), Staphylococcus aureus 18 (8.73%), Pseudomonas aeruginosa 15 (7.28%), Escherichia coli 11 (5.33%), Stenotrophomonas maltophilia 3 (1.45%), Enterococcus species 3 (1.45%), and Serratia species 1 (0.48%). About 95.38% resistance to ceftazidime and cefotaxime and 92.31% resistance to ciprofloxacin and cefepime were found in K. pneumoniae isolates. A. baumannii isolates were 100% resistant to cefotaxime, ceftriaxone, and cefepime. About 22.22% resistance to vancomycin and 66.67% resistance to clindamycin, erythromycin, and cefoxitin were seen in S. aureus isolates. Knowledge of bacterial co-infections and their antibiotic resistance pattern in COVID-19 patients can help in choosing effective antibiotics for the treatment and prevention of antibiotic resistance.

BLOODSTREAM INFECTIONS AND ITS ANTIBIOGRAM OF INTENSIVE CARE UNITS IN A TERTIARY CARE HOSPITAL OF CENTRAL INDIA

Objective: To perform bacteriological analysis and to study drug susceptibility patterns of isolates from bloodstream infections. Methods: This retrospective study was conducted in the microbiology department of NSCB Medical College, Jabalpur, spanning from July 2023 to December 2023. The study included blood culture samples from all adult intensive care unit patients of the medical college. Blood samples were collected with aseptic guidelines and cultured for 7 days. Growths were identified using standard biochemical tests and subjected to sensitivity testing according to Modified Kirby-Bauer’s disk diffusion method. Data for the source of blood collection and duration of incubation were noted and compared. Results: A total of 100 (21.7%) pathogens were isolated from 460 bacteremia suspect patient blood specimens. Pseudomonas spp. Were predominant organisms recovered followed by Klebsiella spp. and Staphylococcus aureus, coagulase-negative staphylococci, and Acinetobacter spp. were the primary pathogens isolated. Carbapenems, glycopeptides, and aminoglycosides were the most effective drugs for treating bacteremia. Conclusions: Early diagnosis and proper antimicrobial therapy lead to successful treatment of sepsis and decreased morbidity and mortality. Antibiogram of a particular area helps in rationalizing verified treatment strategies.

Antibacterial activity of lactic acid bacteria isolated from traditionally fermented food against food pathogen

Aim: To evaluate the antibacterial activity of lactic acid bacteria against selected food pathogens (Escherichia coli and Staphylococcus aureus). Method: A total of twenty (20) traditionally fermented food samples were purchased from a market in Owerri metropolis. Isolation and identification of lactic acid bacteria (LAB) were conducted using standard microbiological techniques. LAB was identified using standard morphological and biochemical tests. They were tested against food pathogens using the Agar well diffusion method. Results: The isolated lactic acid bacteria include Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus plantarum and were label as SP1, SP2, SP3, and SP4 respectively. The isolated LAB species had a considerable inhibitory effect on food borne pathogens. Cell free supernatant of four species were tested for antagonistic activity at 24hrs and 48hrs incubation time. Cell free supernatant from 48hrs broth of SP1 showed the highest zone of inhibition on Escherichia coli (28mm) while the cell free supernatant from 24hrs broth of SP4 showed the lowest zone of inhibition on Staphylococcus aureus (13 mm). Conclusion: The antimicrobial activity increases with time. LAB demonstrated high antimicrobial properties against food borne pathogens. This potential can be employed as starters in food industry to inhibit food spoilage microbes and contaminants.

A Comprehensive Study of Bacterial Etiological Agents in Sterile Body Fluids and Antimicrobial Susceptibility Patterns Among Hospitalized Patients at an Academic Medical Center in India.

Background Sterile body fluids are devoid of any microbial presence, including commensal bacteria. However, bacterial invasion of these fluids can result in life-threatening infections, often leading to significant morbidity and mortality. Timely detection and precise identification of pathogens, along with antimicrobial susceptibility testing, are critical for optimizing therapeuticinterventions and improving patient outcomes. Objective To study the prevalence of bacterial infections in various body fluids in hospitalized patients and to determine the antimicrobial susceptibility patternand the phenotypic detection of extended-spectrum beta-lactamase (ESBL), metallo-beta-lactamase (MBL) and AmpC beta-lactamase producers within bacterial isolates. Materials and methods Sterile body fluid samples, excluding blood and urine, were collected and cultured at the Department of Microbiology, Krishna Institute of Medical Sciences, Western Maharashtra, India, from November 2022 to 2023. The microorganisms isolated from these fluids were identified using standard biochemical tests. Antibiotic sensitivity was assessed through the disc diffusion assay (zone of inhibition) and phenotypic identification of beta-lactamaseenzymes was performed using the combined disc diffusion method. Results During the study period, 180 sterile fluid specimens were collectedrepresenting 48 cerebrospinal fluid (CSF), 53 pleural fluid, 23 peritoneal fluid, and other sterile body fluid samples. Out of these, (n=32, 17.77%) samples were culture-positive. Gram-negative bacteria were oftentimes isolated at 84% (27/32), while gram-positive were 16% (5/32). Escherichia coli was frequently isolated and (n=9, 28.12%) exhibited maximum sensitivity to gentamicin and fosfomycin (n=7, 77.78%) and maximum resistance to cefoperazone-sulbactam (n=8, 88.88%). Pseudomonas aeruginosawas isolated as the second most common organism and showed maximum susceptibility to fosfomycin (n=5, 83.34%) and maximum resistance to gentamicin, cefotaxime, cefoxitin, etc. (n=5, 83.34%). Among gram-positive isolates, coagulase-positive Staphylococcus was high in prevalence rate and (n=3, 9.37%) presented 100% sensitivity to vancomycin and maximum sensitivity to tetracycline(n=2, 66.67%) and 100% resistance to ciprofloxacin, cefoxitin, erythromycin, and other antibiotics. Among gram-negative isolates, MBLproducers were 48.15%, ESBLproducers were 40.74%, and 18.51% were AmpCbeta-lactamase producerswith a multidrug-resistant (MDR) occurrence rate of 93.75%. Conclusion Infections affecting sterile body fluids are critical due to their high mortality and morbidity rates. Timely identification of the causative organisms and their antibiotic susceptibility is essential. The prompt initiation of appropriate antibiotic therapy can decreasethe duration of hospitalization and mitigate the emergence of drug resistance. The presence of MDR organisms in sterile body fluids constitutes considerable challenges in the management of critically ill patients.

Evaluation of blaOXA-48-like point mutation carbapenemase-producing Enterobacterales in Prapokklao Hospital, Thailand.

In this study, we aimed to investigate genetic variation and CPE among blaOXA-48-like carrying isolates recovered from Prapokklao Hospital, Chanthaburi Province, Thailand, during 2016-2017. A total of 122 carbapenem-resistant Enterobacterales (CRE) were recovered from clinical samples in Prapokklao Hospital. All CRE samples were confirmed by standard biochemical tests and minimum inhibitory concentration (MIC) test strips (E-test). The carbapenemase production was determined using the modified Hodge test (MHT), the modified carbapenem inactivation method (mCIM), and EDTA-CIM (eCIM). Three single mutations (E168Q, S171A, and R214S) were characterized in this study. This mutation might reflect the hydrolysis of the modified β-lactam spectrum, especially carbapenem, by OXA-48-like. Our report provides evidence of the blaOXA-48-like point mutation and carbapenemase-producing phenotype of CRE detected in this healthcare setting. Effective control measures and active surveillance of drug resistance in nosocomial pathogens are crucial for controlling diseases associated with difficult-to-treat bacteria.

Bacterial Contamination of Donor Blood Units in a Ghanaian Blood Bank: A Critical Concern for Transfusion Safety

Abstract Introduction/Objective Background: Blood transfusions, while life-saving, pose a significant risk to patient safety, with transfusion-associated bacterial sepsis emerging as a persistent and potentially fatal threat. The prevalence and severity of bacterial contamination in donor blood units necessitate regular surveillance to ensure safe and effective transfusion therapy. Aim This study aimed to identify and assess the susceptibility of bacterial isolates in whole blood donor units at the blood bank of Cape Coast Teaching Hospital in the Central Region of Ghana. Methods/Case Report Method(s): One hundred and three (103) units of stored whole blood units, declared safe for transfusion (testing negative for HIV I&amp;II, HBsAg, HCV, and syphilis), were inoculated into Brain Heart Infusion Broth and incubated for 24 hours. The inoculated broths were then cultured on MacConkey agar (MAC), Chocolate agar (CHOC), and Blood agar. Gram-negative and Gram-positive isolates were identified using standard biochemical tests. Antimicrobial susceptibility testing was performed using the disc diffusion method. Results (if a Case Study enter NA) Results: Of the 103 donor blood units tested, 33 (32.03%) were found to have bacterial isolates. Gram-positive organisms accounted for 67.14% of the isolates, including Coagulase negative Staphylococcus, Bacillus spp, and S. aureus. Gram-negative bacteria constituted 32.86% of the isolates, with Citrobacter freundii, Serratia marcescens, E. coli, and Providencia stuartii identified. Notably, 62.9% of the bacterial isolates were skin commensals, while 25.7% were of enteric origin and 11.4% were of nosocomial origin. Resistance to commonly used antibiotics varied among isolates, with Coagulase-negative Staphylococcus exhibiting resistance to amoxicillin (92.86%) and erythromycin (76.19%), while Citrobacter freundii showed resistance to levofloxacin (75.00%) and ofloxacin (75.00%). Conclusion Conclusion(s): The high incidence of bacterial contamination in donor blood units highlights the need for improved phlebotomy procedures and decontamination practices at blood banks in Ghana to reduce the risk of sepsis in transfusion recipients. Furthermore, this study provides valuable information regarding antibiotic resistance patterns in bacterial isolates, emphasizing the importance of antibiotic stewardship in Ghana. In conclusion, this study underscores the urgency of addressing bacterial contamination in donor blood units in Ghanaian blood banks to enhance transfusion safety.

Detection of virulence factors and antimicrobial susceptibility pattern of clinically significant Klebsiella pneumoniae isolates in a tertiary care hospital

Introduction: Klebsiella pneumoniae is associated with a variety of infections across all age groups, both in community and hospital settings. Many strains of K. pneumoniae produce virulence factors such as a capsule, siderophores, biofilm, and hypermucoviscosity, which facilitate adhesion, invasion, and colonization. Additionally, the rising antibiotic resistance in K. pneumoniae strains poses a significant challenge, limiting treatment options. This study aims to identify the virulence factors and assess the antimicrobial susceptibility of clinically significant K. pneumoniae isolates. Materials and Methods: K. pneumoniae isolates were obtained from urine, sputum, blood, cerebrospinal fluid, pus swabs/aspirates, and tissue samples. Standard biochemical tests were performed for identification. Capsule formation, hypermucoviscosity, siderophore production, and biofilm formation were detected phenotypically. Antibiotic susceptibility testing was carried out using the disk diffusion method, and primary and confirmatory tests were conducted for extended-spectrum beta-lactamases (ESBL) and carbapenemase production. Results: Capsule formation, hypermucoviscosity, siderophore production, and biofilm formation were detected in 100%, 15%, 40%, and 15% of the isolates, respectively. Of the isolates, 40% were multidrug-resistant, 55% produced ESBL, 8% expressed AmpC beta-lactamase, and 10% produced carbapenemase. Conclusion: Capsule formation was the most prevalent virulence factor, followed by siderophore production. The majority of K. pneumoniae isolates demonstrated susceptibility to amikacin, piperacillin-tazobactam, and carbapenems. However, 40% of the isolates were multidrug-resistant, with ESBL production being the most common resistance mechanism.

7010 The Incidence Of Cushing Syndrome In Wisconsin Is More Common Than Population-Based Studies From Europe

Abstract Disclosure: T. Matoska: None. T.B. Carroll: Consulting Fee; Self; Corcept Therapeutics. Research Investigator; Self; Corcept Therapeutics. T.S. Wang: None. S. Dream: None. N. Zwagerman: None. J.W. Findling: Advisory Board Member; Self; Corcept Therapeutics, Recordati, Diurnal. Introduction: Endogenous neoplastic hypercortisolism [Cushing syndrome (CS)] is considered to be a rare disorder. Population-based studies from Europe estimate the incidence of CS to range from 1.8-3.2 cases/million patient-years, and there is limited data reporting the incidence of CS in the United States. All previous studies have shown that pituitary ACTH secreting tumors [Cushing disease (CD)] are the most common cause of CS. Based on our clinical experience, we speculated that the incidence of CS was more common than reported and that adrenal CS (ACS) was more common than CD. This study reports the incidence and etiology of CS from a single tertiary care center in Wisconsin (WI). Methods: Institutional clinic records between May 1, 2017, and December 31, 2022, were queried to identify WI residents treated for CS. Only patients diagnosed and treated at our institution during the study period were included. The diagnosis of CS was established with standard guideline-supported biochemical testing and appropriate imaging. Patients diagnosed with exogenous CS and those who did not undergo therapy for CS were excluded. Results: The query identified 185 patients who were diagnosed and treated for CS. These patients resided in 27 of the 72 WI counties representing a population of 4.5 million of the total WI population of 5.9 million. This suggests that, at a minimum, the incidence of CS in WI is 7.2 cases/million patient-years. Moreover, data from the WI Hospital Association show our institution treated &amp;lt;50% of all patients with CS discharged from WI hospitals from 2019-2023. Mean age at diagnosis was 52.4 years (SD 14.7) and 135 (73%) were female. Of the total cohort, 111 (60%) had ACS, 68 (36.8%) had CD, and 6 (3.2%) had ectopic ACTH syndrome (EAS). Patients with ACS had significantly lower LNSC, DST cortisol, and UFC when compared to those with CD and EAS. ACS had mean (SD) LNSC, DST cortisol, and UFC of 0.236 µg/dL (0.367), 6.5 µg/dL (8.7), and 64.2 µg/24h (92.8), respectively. CD had mean (SD) LNSC, DST cortisol, and UFC of 0.425 µg/dL (0.403), 11.7 µg/dL (7.5), and 151.1 µg/24h (287.4), respectively. EAS had mean (SD) LNSC, DST cortisol, and UFC of 3.189 µg/dL (0.931), 42.5 µg/dL (40.5), and 1514.2 µg/24h (1278.9), respectively. Overt physical features of CS were documented in the medical record of 117 (63.2%) patients at diagnosis ((ACS: 49 (44%); CD: 62 (91%); EAS: 6 (100%)). Conclusion: This study suggests that ACS is more common than CD as a cause of CS, and patients with ACS more frequently presented without physical features of hypercortisolism, probably due to the milder degree of cortisol excess. The incidence of CS in WI is at least 7.2 cases/million patient-years. As our institution cares for &amp;lt;50% of patients with CS in WI, the true incidence is likely far greater and may approach 14-15 cases/million patient-years. Presentation: 6/1/2024

Antimicrobial Susceptibility Patterns of Bacteria Associated with Upper Respiratory Tract Infections in North and South Waziristan, Pakistan

Background: Upper respiratory tract infections (URTIs) are a significant cause of outpatient visits globally, with antimicrobial resistance (AMR) presenting a growing challenge to effective treatment. In Pakistan's tribal regions, limited healthcare access exacerbates this issue.Objective: To determine the prevalence of bacterial pathogens causing URTIs and assess their antimicrobial susceptibility patterns in North and South Waziristan, Pakistan.Methods: This cross-sectional study involved 300 throat swabs collected from outpatients between January and June 2023. Samples were inoculated on Blood, MacConkey, and Chocolate agars, followed by incubation at 37°C for 24 hours. Standard biochemical tests were used for bacterial identification, and the Kirby-Bauer disc diffusion method assessed antimicrobial susceptibility. Statistical analysis was performed using SPSS version 25.0, with p-values ≤ 0.05 considered significant.Results: Pathogens were isolated in 93% of samples, with Staphylococcus aureus (40.2%), viridans group streptococci (35.7%), and Streptococcus pyogenes (17.1%) being the most common. Resistance was highest in viridans group streptococci (33.7% to tazobactam) and lowest in Streptococcus pyogenes (26.3%).Conclusion: The high rates of resistance, particularly in viridans group streptococci, highlight the need for targeted interventions to mitigate AMR in this region.