Standard Microbiological Techniques Research Articles

Correlation of procalcitonin and c-reactive protein levels with pathogen distribution and infection localization in urinary tract infections

Aimed to explore the relationships between infection localization, bacterial species, and procalcitonin (PCT) and C-reactive protein (CRP) levels in urinary tract infections (UTIs). A retrospective study included 314 UTI hospitalized patients divided into two groups (268 with lower UTI, 46 with upper UTI) in a tertiary care hospital. PCT and CRP were performed. Bacterial isolates were identified using standard microbiological techniques, and statistical analyses were performed to assess associations between infection localization, bacterial species, PCT, and CRP levels. Age and gender showed no significant differences between the lower and upper UTIs. Escherichia coli dominated as the leading UTI pathogen. A positive correlation (r = 0.646, P < 0.001) between PCT and CRP levels was found. The subgroup with ureteritis in the upper UTI category exhibited the highest PCT and CRP levels. PCT and CRP exhibited favorable diagnostic potential in predicting upper UTIs, with AUCs of 0.644 and 0.629, respectively. The optimal cutoff values were 0.21 ng/mL for PCT and 60.77 mg/L for CRP. Sensitivities were 69.03% and 77.99%, while specificities were 56.52% and 47.83%, respectively. E. coli emerged as the predominant bacterium in UTIs. PCT and CRP demonstrated moderate diagnostic efficacy in distinguishing between upper and lower UTIs. Notably, PCT and CRP exhibited enhanced utility in identifying ureteritis.

A Study of Spontaneous Bacterial Peritonitis in Cirrhosis of Liver with Ascites at a Tertiary Care Hospital in North India: Prevalence, Clinical, and Microbiological Profile

Background and Objectives: In patients with chronic liver disease and ascites, spontaneous bacterial peritonitis (SBP) is a common complication. This can progress gradually and subtly or go undiagnosed until signs start to show. Early diagnosis is essential for effective treatment and the prevention of subsequent episodes because the mortality rate following a single episode varies from 20% to 40%. The goal was to study the incidence, clinical profile, and microbiological characteristics of SBP and its variants in people with liver cirrhosis and ascites. Materials and Methods: This observational study was done on 104 cirrhotic patients with ascites admitted at a tertiary care hospital. Basic demographics, symptoms, and clinical signs of patients were recorded. Diagnostic paracentesis was done aseptically and immediately sent for ascitic fluid cytology, biochemistry, and culture. Bacterial examination and antibiotic sensitivity tests were carried out by standard microbiological techniques. Results: Out of a total of 104 cirrhotic patients, 28 (26.9%) were diagnosed with SBP. Alcohol was the most common etiological factor, followed by hepatitis C infection. Among the SBP patients, the most common presentation was culture-negative neutrocytic ascites (71.4%), followed by classical SBP (17.8%), then mono bacterial non neutrocytic bacterascites MNB (10.7%). Among the symptoms, the most common was abdominal distension, followed by icterus, pedal edema, and abdominal pain. Majority of the patients belonged to Child–Pugh’s Grade C. Of 28 cases of SBP, 8 were culture positive. Gram-negative isolates were predominant (75%) and Escherichia coli were the most common isolate. Interpretation and Conclusion: The prevalence of SBP in cirrhotic patients was 26.9%, and Gram-negative isolates were predominant. Ascitic fluid culture and susceptibility testing can lead to accurate diagnosis of SBP and can guide for treatment as resistance to antibiotics is common.

Dissemination of Multidrug-Resistant, Extensively Drug Resistant and Pandrug-Resistant Pseudomonas aeruginosa Isolates among In-Patients and Out-Patients in a Multi-Profile Health Care Settings

Pseudomonas aeruginosa is one of the most life-threatening pathogens, especially in healthcare settings, and a main contributor to multi-drug resistance (MDR), extensive-drug resistance (XDR), and pan-drug resistant (PDR) phenotypes. However, there is limited data on the degree of resistance of these isolates in this region. This study seeks to determine the distribution of MDR, XDR, and PDR Pseudomonas aeruginosa strains from different patient groups. A total of five hundred (500) non-duplicated strains of Pseudomonas aeruginosa of human clinical samples were collected from the Microbiology Laboratory Unit of Alex Ekwueme Teaching Hospital Abakaliki. The isolates were identified and re-characterized by standard microbiology techniques. MDR, XDR, and PDR were determined using the Kirby–Bauer disk diffusion method, and the results were analyzed using the Clinical Laboratory Standard Institute (CLSI) zone diameter breakpoints. The result shows that the overall resistant phenotype was MDR 50. 7%, XDR 20.5%, and PDR 9.6% while in samples from in-patients and out-patients, resistant phenotype proportions were MDR 43.2%, XDR 32.4%, PDR 10.1% and MDR 61.2%, XDR 29.7%, and PDR 18.5% respectively. Worrisomely, only a few tested antimicrobial agents (Amikacin, cefepime) were active against the test organism, presenting a limited therapeutic option. It is therefore imperative to establish strong regulative measures and guidelines that would help in curtailing the increasing dissemination of these superbugs in healthcare institutions in Nigeria.

Occurrence of Virulence Genes among Methicillin-Resistant Staphylococcus aureus Isolated from Subclinical Bovine Mastitis.

The occurrence of Staphylococcus aureus-induced subclinical mastitis holds significant implications for public health. This specific microorganism possesses a wide array of pathogenic factors that enable it to adhere to, colonize, invade, and infect the host. The objective of the current study was to assess the prevalence of S. aureus, determine antimicrobial resistance patterns, and identify virulence genes of methicillin-resistant S. aureus (MRSA) strains responsible for subclinical mastitis in bovines. A total of 249 milk samples were collected from various farms in the district of Faisalabad. The presence of subclinical mastitis was assessed by using the California mastitis test. Positive milk samples (n = 100) were then subjected to standard microbiological techniques for isolation and identification of S. aureus. Antibiogram analysis was conducted by using the disc diffusion method to assess antimicrobial resistance. For the molecular detection of S. aureus and its virulence genes, the polymerase chain reaction (PCR) was performed with species-specific primers. The overall prevalence of S. aureus was found to be 40% (40/100), which was confirmed through molecular detection of the nuc gene in 40/40 (100%) of samples using PCR. Antimicrobial susceptibility tests indicated the highest susceptibility to vancomycin, sulfamethoxazole/trimethoprim, erythromycin, gentamicin, ciprofloxacin, and chloramphenicol, while the highest resistance rate was observed against tetracycline. Additionally, 30% of samples (12/40) tested positive for methicillin resistance. PCR analysis revealed that 100% of MRSA-tested isolates harbored the mecA and clfA genes. Furthermore, the MRSA isolates showed the presence of pvl, hla, hlb, sec, icaA, icaD, icaB, and icaC genes at rates of 92, 75, 67, 42, 42, 75, 8, and 25%, respectively. These findings underscore the need for stricter aseptic control in dairy farms to prevent disease transmission between animals and ensure the production of safe and uncontaminated food for human consumption.

Comparative inhibitory effects of sodium and potassium chloride salts on Staphylococcus aureus from fermented Pentaclethra macrophylla

Aim: The study was conducted to evaluate comparatively the individual effects of sodium (NaCl) and potassium (KCl) chloride salts against Staphylococcus aureus isolated from fermented Pentaclethra macrophylla. Method: A total of twenty (20) traditionally fermented food samples (Pentaclethra macrophylla) were purchased from a market in Umuahia metropolis. Isolation and identification of Staphylococcus aureus were conducted using standard microbiological techniques. Staphylococcus aureus was identified using standard morphological and biochemical tests. The inhibitory effect of both salts (NaCl and KCl) were analyzed on modified nutrient agar and nutrient broth (Media + 5% salt conc., Media + 10% salt conc., Media + 15% salt conc., Media + 20% salt conc.). The study measured culture growth using optical density readings at 425nm over 48 hours with different NaCl and KCl concentrations, while growth on plates was visually assessed on colony counter. Results: The highest optical density recorded as a result of contact with varying concentration of sodium chloride was 0.48 at 5% concentration after 48hours incubation while the least was 0.04 at 20% concentration after 12hours incubation while the highest optical density recorded as a result of contact with varying concentration of potassium chloride was 0.41 at 5% concentration after 48hours incubation while the least was 0.01 at 20% concentration after 12hours incubation. Conclusion: Higher salt concentrations resulted in osmotic shock, suppressing growth in Staphylococcus aureus, but increased salt concentration partially alleviated growth inhibition over time.

ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF COMMON BACTERIAL UROPATHOGENS, A COMPREHENSIVE STUDY AT TERTIARY CARE HOSPITAL OF BALOCHISTAN

Objective: To assess the frequency of the bacterial agents responsible for UTIs and their antibiotic susceptibility patterns in a tertiary care hospital. Material and Methods: This Laboratory- based cross- sectional study was conducted at The Microbiology Department, Combined Military Hospital, Quetta in time duration of one year from April 2022 to March 2023. This study encompassed 512 bacterial isolates that were cultured positive from a collection of 1282 urine samples obtained from patients receiving care both in indoor and outdoor settings. The specimens were inoculated on CLED agar. Identification of isolated colonies was done by standard microbiological techniques and the Kirby Bauer disc diffusion method was used for antibiotic susceptibility testing. Quality control was ensured by using reference strains, and antibiotics were tested according to the CLSI 2022 recommendations. Results: Bacterial uropathogens were identified in urine samples of 33.5% of male and 44.7% of female patients, with an overall infection rate of 39.93%. Of the 512 bacterial isolates, 91.79% were Gram-negative and 8.21% were Gram-positive. Escherichia coli was the most common isolate (69.53%), followed by Klebsiella pneumoniae (12.1%), Enterococci (7.6%), Pseudomonas (2.92%), among other bacterial species. The gram-negative isolates were sensitive to Nitrofurantoin (96%), Fosfomycin (93%), Amikacin (79%), Imipenem (75%), and Meropenem (74%), while gram positive isolates were highly sensitive to Linezolid (100%), Vancomycin (95%) and Nitrofurantoin (90%). Conclusion: E. coli was the most common cause of UTIs at our hospital, followed by Klebsiella pneumonia with Nitrofurantoin and Fosfomycin being effective treatment options. Keywords: E coli, Nitrofurantoin, Uropathogens.

Comparative Effects of Prepared and Vended Herbs on Selected Diarrhoegenic Bacteria in Ekiti State

Herbs contamination by some pathogenic bacteria poses a major threat to the usage as a treatment option for diarrheagenic bacteria. This study was carried out to determine the comparative effects of prepared and vended herbs sold within Ado-Ekiti metropolis on selected diarrhogenic bacteria. Herb was prepared in the laboratory under a strict aseptic condition, and a total of 50 vended herbs samples were purchased from vendors across ten different locations within Ado-Ekiti, Ekiti State. All herb samples were analyzed using standard microbiological techniques, total bacteria count, Staphylococcal counts, coliform counts and fungal counts were carried out. Bacteria colonies on the agar plates were identified using morphological and biochemical characterization. The susceptibility testing of the vended and prepared herbs on selected diarrheagenic bacteria; Shigella dysenteriae, Escherichia coli and Salmonella typhi were carried out using agar well diffusion method. The haematological and histological effects of the herbs were done using standard haematological and histological procedures respectively. The data generated were analysed using analysis of variance (ANOVA) and the level of significance was set at 0.05. There was a low microbial count in the prepared herb compared to the vended herbs, Staphylococcal counts, coliform counts and fungal counts of the vended herbs ranged from 15.50±0.65 to 52.00±0.36 cfu/100ml, 7.00±0.40 to 22.00±0.13 cfu/100ml, 8.50±0.15 to 36.50±0.20 cfu/100ml and 1.50±0.50 to 8.00±0.40 sfu/100ml respectively while the total bacterial counts of prepared herbs ranged from 1.00±0.01 to 2.00±0.00 cfu/100ml. There was a low microbial count in the herbs vended by those with more than 9 years herb vending experiences compared to others. Nine different bacteria species were isolated from vended herb samples with Escherichia coli and Staphylococcus aureus 8(22.2%) being the most frequently occurred bacteria and followed by Enterobacter aerogenes and Pseudomonas aeruginosa was 4(11.11%) occurrence. The laboratory prepared herbs exhibited a better antibacterial activity against the tested diarrheagenic bacteria (S. dysenteriae (6.00±0.00 mm), E. coli (10.00±0.10 mm) and S. typhi (8.00±0.40 mm) compared to the vended herbs. The bacteria isolated from the vended herbs showed higher resistant against augmentin and amoxicillin. The haematological profile of the rats showed a significant increase in the white blood cell counts. There were also multiple foci of hepatic necrosis with cellular aggregation around the same foci in the liver and diffuse tubular degeneration and necrosis with the tubular epithelium completely eroded in the kidney of the rat. However, occurrence of these bacteria in vended herbs could predispose consumers to the risk of infections with pathogenic and antimicrobial resistant bacteria. Also, the herbs could be a source of toxicity showed in both histological and haematological parameters, which could pose a life-threatening disease to the consumer. Therefore, steps involve in the preparation of the herb must be standardize to prevent the microbial contamination.

Fecal carriage of extended spectrum beta-lactamase and fluoroquinolone resistant gene in non-typhoidal Salmonella enterica isolates from food-producing animals and humans

This study seeks to determine the feacal carriage of extended spectrum beta-lactamase and fluoroquinolone resistant non-typhoidal Salmonella enterica isolates from food-producing animals and humans. A total of three hundred (300) fecal samples were collected using sterile universal containers from food-producing animals namely (Chicken [100], Pig [100] and humans (100) from Onicha Local Government Area of Ebonyi State and analyzed for the presence of non-typhoidal Salmonella enterica using standard microbiological techniques. Phenotypic detection of extended-spectrum beta-lactamase (ESBL) were done by disc diffusion and Double Disk Synergy Test. Molecular characterization for ESBL and fluoroquinolone-resistant genes were done by PCR with specific primers. The result shows that non-typhoidal Salmonella species (NTS) accounted for 25 % and 17 % in poultry and pig fecal sample respectively while 60 % and 40% were phenotypic ESBL producers respectively. When compared statistically there is significant difference among isolates confirmed ESBL-positive (P˂ 0.05). Also, none of the 16 (58 %) NTS isolated from humans harbored ESBL phenotype. PCR analysis with β-lactam specific primer detected the presence of blaOXA 50 % and 50 %, blaSHV 36 %, and 64 %, blaTEM 43 % and 57 %, blaCTX-M 36 % and 64 % in poultry and pig respectively. Fluoroquinolone resistant gene QnrA was present in 0 and 100 % of poultry and pig respectively. QnrB was 40 % and 60 % in poultry and pig isolates respectively. QnrS was present in 64 % isolates of poultry and 13 % isolates in pig. The high prevalence of genes encoding beta-lactamases and fluoroquinolone resistance (TEM, SHV, CTX-M and OXA, (qnrA, qnrB and qnrS) were present more in poultry and pig than in humans and demonstrate a significant public health threat from consumption of food-producing animal harboring such pathogenic resistant genotype if not properly controlled.&#x0D; Keywords: Extended spectrum beta-lactamase, Fluoroquinolone, non-typhoidal Salmonella enterica, feacal carriage

Bacterial Examination in Spoiled Vegetables, Its Isolation, Identification and Antibacterial Activity of Turmeric, Pongamia and Betel Leaf

Microbial contamination is a major concern in the food and pharmaceutical industries, leading to economic losses and public health risks. Natural products such as turmeric, betel leaf, and Pongamia leaves have been traditionally used for their antimicrobial properties. This thesis aimed to examine, identify, and isolate bacteria from selected spoiled vegetable samples and to evaluate the antibacterial activity of turmeric, betel leaf and Pongamia leaves. Bacterial isolation was performed using standard microbiological techniques, and the identification was done based on cultural characteristics. The antibacterial activity was assessed using disc diffusion method. The results showed the presence of bacterial contaminants in the tested samples, including Staphylococcus sp. And Escherichia coli. The turmeric, betel leaf, and Pongamia leaves extracts exhibited significant antibacterial activity against the tested bacteria. The phytochemical analysis of the extracts revealed the presence of alkaloids, flavonoids, phenols, tannins, and saponins, which may contribute to their antibacterial properties. This research provides valuable insights into the microbial contamination of food and pharmaceutical samples and the potential of natural products as antimicrobial agents. The findings of this study can contribute to the development of new antimicrobial agents based on natural products and the improvement of food and pharmaceutical safety.

Prevalence, molecular characterization of integrons and its associated gene cassettes in Klebsiella pneumoniae and K. oxytoca recovered from diverse environmental matrices

The high prevalence of infections arising from Klebsiella species is related to their ability to acquire and disseminate exogenous genes associated with mobile genetic elements such as integrons. We assessed the prevalence, diversity, and associated gene cassettes (GCs) of integrons in Klebsiella species. The isolates recovered from wastewater and hospital effluents, rivers, and animal droppings were identified using the conventional Polymerase Chain Reaction (PCR) with primers targeting the gryA, pehX, and 16S–23S genes. The antimicrobial resistance profile and the Extended-Spectrum and Metallo β-lactamases production were carried out using standard microbiological techniques. PCR, DNA sequencing analyses, and Restriction Fragment Length Polymorphism were used to characterize the integrons and their associated GCs. Furthermore, the genotypic relationships between the different isolated K. pneumoniae were determined using Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR. About 98% (51/52) of the confirmed isolates harboured an integrase gene, with 80% intI1, while the remaining 20% concurrently harboured intI1 and intI2, with no intI3 observed. About 78% (40/51) of the bacterial strains were positive for a promoter, the P2R2, investigated, while 80% (41/51) harboured at least one of the qacEΔ1 and sul1. Three different GCs arrangements identified were aac(6′)-Ib, aadA1-dfrA1, and dfrA1-sat2. At a similarity index of 60%, the ERIC-PCR fingerprints generated were categorized into nine clusters. Our study is the first to reveal the features of integrons in Klebsiella spp. recovered from environmental sources in the Eastern Cape Province, South Africa. We conclude that the organisms' sources are repositories of integrons harbouring various gene cassettes, which can be readily mobilized to other microorganisms in similar or varied niches.

Antimicrobial Susceptibility Pattern of Bacterial Isolates in Patients with Lower Respiratory Tract Infections

Lower respiratory tract infection (LRI) is one of the global public health problems accounting for millions of deathsannually. The present study aim is to determine bacterial pathogens' bacteriological profile and antibiotic susceptibility patternaffecting the patients with LRI attending a tertiary care hospital. Two hundred and eight sputum samples were collected frompatients with signs and symptoms of LRI. The samples were processed in Microbiology laboratory for bacterial pathogens.Bacterial isolates were identified using standard microbiological techniques and antibiotic susceptibility testing was performedusing the Kirby-Bauer disc diffusion method. Klebsiella pneumoniae (39.6%), followed by Pseudomonas (32%), were the mostpredominant bacteria isolated in the present study. The other bacteria isolated were Escherichia coli (16.9%), Staphylococcusaureus (5.6%), Streptococcus species (1.8%), and Acinetobacter (3.7%). Gram-negative bacteria showed high drug resistance toampicillin and cephalosporins and maximum susceptibility to piperacillin-tazobactam, amikacin, and gentamicin. Most of theisolates showed resistance to the Beta-lactam group of antibiotics. The isolated bacterial pathogens' resistance profile indicatesthe need for the appropriate diagnosis and antibiotic susceptibility testing before managing the cases.

Antimicrobial Activity of Acacia Nilotica and Ziziphus Mauritiana against Clinical Isolates of Escherichia Coli and Klebsiella Aerogenes

Klebsiella aerogenes is an opportunistic pathogen linked to several nosocomial illnesses, including pneumonia, sepsis, and infection of the urinary tract. Escherichia coli is frequently the source of numerous common bacterial illnesses, including bacteraemia, cholecystitis, cholangitis, urinary tract infections (UTI), traveler's diarrhoea, and other medical conditions such as meningitis in babies and pneumonia. This study aimed at determining the antimicrobial activity of Acacia nilotica and Ziziphus mauritania on clinical isolate of E. coli and K. aerogenes. The phytochemical constituents of Z. mauritiana and A. nilotica were determined. E. coli and K. aerogenes isolates were obtained from the Barau Dikko Teaching Hospital, Kaduna and reconfirmed using standard microbiological techniques. The antibacterial activity of Z. mauritiana and A. nilotica extracts against the isolates was determined using Agar well diffusion assay, the MIC and MBC was also determined and recorded. The findings of this study revealed that tannins, quinone, phenols, terpenoids, and steroids are present in Z. mauritiana extract while tannins, saponin, quinones, and terpenoids in A. nilotica extract. There was no activity of Z. mauritiana against any of the test isolates at all concentrations used in this study. Whereas A. nilotica exhibited an antibacterial activity against both E. coli and K. aerogenes recording respectively a zone of inhibition of 24 mm and 25 mm with MIC and MBC value of 600 mg/ml. This research displayed an antibacterial activity of A. nilotica and no activity of Z. mauritiana against E. coli and K. aerogenes.

Microorganisms, proximate analysis, nutritional contents and amino acids profile of naturally opened ackee apple fruits (Blighia sapida)

Fruits are important foods that contain nutrients which contribute to the growth and mental health. A total of 100 naturally opened Ackee apple (Blighia sapida) fruits were collected in Ado-Ekiti. The samples were analyzed using standard microbiological techniques examination. The proximate analysis, nutritionally viable minerals contents, and functional properties of the amino acid profile were determined. Fungal isolated were Asperrgillus niger (23 %), Mucor species (11 %), Rhizopus species (15 %) and Aspergillus flavus (35 %) while Bacterial isolates were Bacillus laterusporus (7 %), Staphylococcus aureus (5 %) and Pasteurella sp. (4 %). The sources of microorganisms that contaminated these fruits may include air, birds, insects, rabbits, rats and other animals. The proximate analysis of Ackee Apple Fruits were Ash(3.62 %), Moisture content (40.23%), Crude Protein (5.5 %), Fat (11.55 %), Fibre (3.49 %) and Carbohydrate (35.63%); Mineral contents were Sodium (201.49 mg/100 g), Potassium (652.91 mg/100g), Calcium (8.24 mg/100g), Zinc (258.68 mg/100 g), Magnesium (20.86 mg/100 g), Manganese (0.05 mg/100 g), Iron (7.87 mg/100g) and Phosphorus (48.6 mg/100g); Functional properties were oil absorption (144.5 W/V), Water absorption (378.06 W/V), Foaming capacity (14.50 W/V) and Least gelation concentration (6 W/V) while the amino acid contents ranged from 16 to 132 g/100 g. Crude protein Glutamic acid was the highest Amino acid while Tyrosine was the least. Ackee apple fruits provide nutritive values for health since it contains essential minerals. The health implications of isolated microorganisms are that they can cause mycotoxin and cause food-borne diseases. It is imperative for the consumers of Ackee apple fruits should ensure proper washing and maintain a high standard of hygiene.

BIO-AEROSOLS AND THEIR VIRULENCE FACTORS IN EDONWHII BEACH ATMOSPHERE, EASTERN OBOLO LGA, NIGERIA

Standard microbiological techniques were used to evaluate bio-aerosols and associated virulence factors in the Edonwhii Beach Atmosphere in Eastern Obolo LGA, Nigeria. The microbial components of the beach atmosphere were evaluated using Settle Plate and Metagenomics methods. All the culturable bacteria but Escherischia coli, elaborated virulence factor(s), indicating potential to cause disease. At the molecular level, pathogenic genes fliC and hlyA that code for flagellin C and hemolysin A virulence factors were detected. Also detected were strains of Fusarium verticilliodes, Candida krusei, Candida albicans, Trichophytosporon sp and Cladosporium sp and are capable of causing keratitis or other dermal infections in human. High concentrations of microbial pollutants in Edonwhii beach atmosphere are a clear evidence of air pollution which can adversely impact on human health. As a result, continual monitoring of air quality, as well as devising and developing instruments to identify contaminants, is advised. Open defecation near the beach should be avoided, and overall environmental cleanliness should be maintained.

Antibiotic resistance and virulence genes profiling of Vibrio cholerae and Vibrio mimicus isolates from some seafood collected at the aquatic environment and wet markets in Eastern Cape Province, South Africa.

The current study determines the density of Vibrio spp. and isolates V. cholerae and Vibrio mimicus from fish-anatomical-sites, prawn, crab and mussel samples recovered from fish markets, freshwater and brackish water. Virulence and antibiotic resistance profiling of isolates were carried out using standard molecular and microbiology techniques. Vibrio spp. was detected in more than 90% of samples [134/144] and its density was significantly more in fish than in other samples. Vibrio. cholerae and V. mimicus were isolated in at least one sample of each sample type with higher isolation frequency in fish samples. All the V. cholerae isolates belong to non-O1/non-O139 serogroup. One or more V. cholerae isolates exhibited intermediate or resistance against each of the eighteen panels of antibiotics used but 100% of the V. mimicus were susceptible to amikacin, gentamycin and chloramphenicol. Vibrio cholerae exhibited relatively high resistance against polymyxin, ampicillin and amoxicillin/clavulanate while V. mimicus isolates exhibited relatively high resistance against nitrofurantoin, ampicillin and polymixin. The multiple-antibiotic-resistance-index [MARI] for isolates ranges between 0 and 0.67 and 48% of the isolates have MARI that is >0.2 while 55% of the isolates exhibit MultiDrug Resistance Phenotypes. The percentage detection of acc, ant, drf18, sul1, mcr-1, blasvh, blaoxa, blatem, blaoxa48, gyrA, gyrB and parC resistance-associated genes were 2%, 9%, 14%, 7%, 2%, 25%, 7%, 2%, 2%, 32%, 25% and 27% respectively while that for virulence-associated genes in increasing other was ace [2%], tcp [11%], vpi [16%], ompU [34%], toxR [43%], rtxC [70%], rtxA [73%] and hyla [77%]. The study confirmed the potential of environmental non-O1/non-O139 V. cholerae and V. mimicus to cause cholera-like infection and other vibriosis which could be difficult to manage with commonly recommended antibiotics. Thus, regular monitoring of the environment to create necessary awareness for this kind of pathogens is important in the interest of public health.

Fecal Carriage of Multi-drug Resistant Escherichia coli in Healthy Children: a Community-based Study

Introduction: Escherichia coli is enteric Gram-negative bacilli, earlier recognized as non-invasive commensal. Invasive strains of E. coli have been identified and are associated with a variety of human infections. Increasing drug resistance among E. coli has been reported. Multidrug resistance is the ability of an organism to resist at least one agent in three or more antimicrobial categories. The aim of this study was to detect fecal carriage of MDR E. coli isolates among healthy school children.&#x0D; Methods: A total of 139 stool samples were collected from Rastriya Secondary School, Purano Tudikhel, Pokhara-1. Isolation, identification, and Antibiotic susceptibility test pattern of E. coli isolates were done by standard microbiological techniques. Further Extended Spectrum Beta-Lactamase test was performed by Combined Disc Test and data analysis was done by using Microsoft Excel.&#x0D; Results: A total of 106 E. coli were isolated from the stool samples. MDR E. coli were found to be 8 (7.55%) and 7 E. coli isolates were resistant to Ceftazidime which was further tested for ESBL and 5 isolates were found to be ESBL producers. The resistance of E. coli to Ampicillin, Nitrofurantoin, Ceftriaxone, Ceftazidime, Ciprofloxacin, and Gentamicin was 27.36%, 43%, 10.38%, 6.60%, 6.60%, and 2.83% respectively. Only one isolate was resistant to all antibiotics used except Imipenem.&#x0D; Conclusion: E. coli is the main Gram-negative bacteria isolated from the stool sample and identified as the main carrier of antimicrobial resistance in species comparison of Enterobacteriaceae isolated from fecal samples. Therefore, quick surveillance is needed in order to effectively handle such MDR strains. The E. coli population susceptible to all antibiotics used was a much more diverse group than the resistant and ESBL-producing E. coli.

Pseudodesulfovibrio thermohalotolerans sp. nov., a novel obligately anaerobic, halotolerant, thermotolerant, and sulfate-reducing bacterium isolated from a western offshore hydrocarbon reservoir in India

ObjectiveCharacterization and documentation of strain MCM B-1480T, a novel sulfate-reducing bacterium isolated from produced water of India's western offshore hydrocarbon reservoir. MethodStrain MCM B-1480T was unequivocally identified using a polyphasic approach routinely followed in bacterial systematics. The morphological and biochemical characterization of strain MCM B-1480T was carried out using standard microbiological techniques. ResultsMCM B-1480T was a Gram-stain-negative, motile, non-spore-forming, curved-rod-shaped bacterium. MCM B-1480T could grow at temperatures between 20 and 60 °C (optimum 37 °C), pH 6–8 (optimum 7), and required 1–6% NaCl (optimum 3%) for growth. Strain MCM B-1480T was reducing sulfate to produce hydrogen sulfide during growth. This strain used lactate and pyruvate as prominent electron donors, whereas sulfate, sulfite, thiosulfate, and nitrate served as electron acceptors. MCM B-1480T shared maximum 16S rRNA gene sequence homology of 98.65% with the members of the genus Pseudodesulfovibrio. The G + C content of the 3.87 Mb MCM B-1480T genome was 60.39%. Digital DDH (27.7%) and average nucleotide identity (ANI 84%) with the closest phylogenetic affiliate (less than 70% and 95%, respectively) reaffirmed its distinctiveness. The major cellular fatty acids components, namely iso-C15:0, anteiso-C15:0, C16:0, and anteiso-C17:0, differentiated strain MCM B-1480T from other species of Pseudodesulfovibrio. Genome annotation revealed the presence of genes encoding dissimilatory sulfate reduction and nitrate reduction in strain MCM B-1480T. ConclusionThe polyphasic studies, including SSU rRNA gene sequencing, average nucleotide identity, Digital DNA-DNA hybridization, cell wall fatty acids analysis, etc., identified strain MCM B-1480T as a novel taxon and Pseudodesulfovibrio thermohalotolerans sp. nov. was proposed (= JCM 39269T = MCC 4711T).

Antimicrobial susceptibility and molecular characteristics of beta-lactam- and fluoroquinolone-resistant E. coli from human clinical samples in Nigeria

This study determined the antimicrobial resistance profiles and molecularly characterized beta-lactam- and fluoroquinolone-resistant E. coli recovered from human clinical samples. Three hundred (300) clinical samples collected from two major tertiary healthcare institutions were analyzed using standard microbiological techniques. Antimicrobial susceptibility testing and phenotypic detection of extended-spectrum beta-lactamase (ESBL) were done by disc diffusion and Double Disk Synergy Test. Molecular characterization for ESBL and fluoroquinolone-resistant genes were done by PCR with specific primers. In total, 44 (14.7%) ESBL-producing E. coli were recovered from human clinical specimens. The recovered isolates were multidrug-resistant with multiple antibiotics resistance index values ranging from 0.5–0.8. Isolates exhibited resistance to sulfamethoxazole/trimethoprim 44 (100%), cefoxitin 42 (95.5%), tetracycline 42 (95.5 %), cefuroxime 41 (93.2%), ciprofloxacin 40 (90.9%), cefotaxime 37 (84.1%), amoxicillin 37 (84.1%), amoxicillin/clavulanic acid 37 (84.1), and ceftazidime 35 (79.5%). BlaTEM 41 (93.2%) ESBL gene was the most predominant gene among the isolates, followed by blaCTX-M 9 (20.5%), and blaSHV 1 (2.3%) genes. Co-existence of blaTEM +blaSHV, blaTEM + blaCTX-M, and blaTEM +blaSHV + blaCTX-M genes was observed in 1 (2.3%), 7 (15.9%), and 4 (9.1%) isolates, respectively. All the fluoroquinolone-resistant isolates harbored aac-lb-6-cr gene while Qnr gene was absent. Association of fluoroquinolone resistance- and β-lactam resistance genes; aac-lb-6-cr + blaTEM + blaSHV + blaCTM was observed in 5 (11.4%) isolates. Our study showed high frequency of ESBL-producing E. coli harboring ESBL and fluoroquinolone resistance genes from human clinical samples in Ebonyi State, Nigeria. The high frequency of multidrug-resistant E. coli in our study is very worrisome and could have significant public health impact such as treatment failures, and possibly death, if not properly managed. It is therefore imperative to establish strong regulative measures and guidelines that would help in curtailing the increasing dissemination of these superbugs in healthcare institutions in Nigeria.

DETECTION OF EXTENDED SPECTRUM BETA LACTAMASES-PRODUCING KLEBSIELLA PNEUMONIAE ISOLATES IN CHRONIC OBSTRUCTIVE PULMONARY DISEASES PATIENTS

Objective: Chronic obstructive pulmonary disease (COPD) is the leading cause of morbidity and mortality worldwide which includes chronic bronchitis and emphysema. 80% of acute exacerbation of COPD (AECOPD) are triggered by respiratory viruses, atypical bacteria, and aerobic Gram-positive and Gram-negative bacteria. The aim of our study was to analyze the prevalence and antimicrobial susceptibility testing of Klebsiella pneumoniae and detection of extended spectrum beta lactamases (ESBL)-producing K. pneumoniae isolates from COPD patients. Methods: Sputum samples received from seventy-five diagnosed cases of COPD from the department of General Medicine and Chest Medicine, TSRM MCH and RC, Trichy, from March 2022 to August 2022. Direct Gram stain was done for all sputum samples. The suitable sputum samples were cultured. The identification of organism and antimicrobial susceptibility testing was done by standard microbiological techniques. Results: Out of seventy-five cases, 64% were males and 36% were females. Forty-seven were sputum positive, and the prevalence of Gram- negative bacteria was 64%. K. pneumoniae was the most common bacteria isolated (42.2%), followed by Pseudomonas aeruginosa (15.5%). Antimicrobial susceptibility testing of K. pneumoniae was sensitive to imipenem (78.94%), aminoglycosides (78.94%), amoxycillin-clavulinic acid (63.15%), ciprofloxacin (57.89%), Piperacillin-Taxobactam (57.89%), and the phenotypic tests showed 13 (68%) isolates were ESBL producers. Conclusion: This study showed that K. pneumoniae and P. aeruginosa are the most common organisms associated with AECOPD and increased rate of ESBL producers were observed.

Evaluation of Fungal Contamination on Livestock Feed Ingredients of Plants origin Stored for Five Years

The purpose of this experiment was to evaluate the extent of Fungal contamination of livestock feed stuffs of plants origin stored for five years at Plateau State College of Agriculture, Garkawa. The feed ingredients were: wheat offals, cowpea husks, cotton seeds, kidney beans seeds, baobab seeds, bambara nut husks, palm kernel cake, baobab seed meal, rice offals, yam peels, ground nut cake and sweet orange peels. Microbiological quality of these feedstuffs were conducted using standard microbiological technique for food and animal feeding stuffs at Microbiology Unit of Central Diagnostic Laboratory at National Veterinary Research Institute Vom, Plateau State, Nigeria. The study revealed a total of eight fungi spp: Aspergillus niger, Aspergillus fumigatus, Penicillum spp, Rhizopus spp, Mucor spp, Trichophyton rubrum, Aspergillus spp and Aspergillus flavus. From the findings, cowpea husks, cotton seed, kidney beans seed, baobab seed and bambara nut husks were contaminated by Aspergillus niger (28.57%) as the dominant species of fungi followed by: Aspergillus fumigatus (19.05%) (baobab seed, bambara nut husks, blood meal, and baobab seed meal) and baobab seed (Aspergillus niger, Aspergillus fumigatus and Penicillium spp). Wheat offals and palm kernel cake had the highest total fungi counts of 4.0 x 102 cfu/g followed by yam peels, sweet orange peel and bone meal with 3.0 x 102 cfu/g and baobab seed meal had 2.2 x 102 cfu/g. To reduce the growth of mould on livestock feed ingredients stored over long periods of time, regular monitoring of moisture levels of these ingredients should be checked before embarking on storage.