Multidrug Resistance Index Research Articles

ANTIBIOTIC SUSCEPTIBILITY PATTERN OF BACTERIA ASSOCIATED WITH CLINICAL WASTE MATERIALS IN GOKANA LOCAL GOVERNMENT AREA OF RIVERS STATE.

A significant issue in world healthcare today is the rise in illnesses brought on by bacteria that have developed resistance to widely used antibiotics. The goal of this study was to determine the pattern of antibiotic susceptibility of bacteria that were isolated from five clinical wastes that were gathered from approved health facilities in Rivers state's Gokana Local Government. Fifty (50) samples of clinical waste were collected for six months from two different health centers. Samples were subjected to Standard microbiological analysis. Total heterotrophic count, total Staphylococcal count, coliform count, and feacal coliform count were estimated. Five (5) bacteria species isolated during the study and their percentage occurrence were Staphylococcus sp. was the most frequently isolated bacteria 15 (55.6%), Bacillus sp. 8(29.6%), Klebsiella sp. 2(7.4%), Pseudomonas sp. and Escherichia coli 1 (3.7%). Susceptibility of bacteria to antibiotics revealed that the Gram-positive bacteria were 77.3% resistant to Cefexime, 68.2% resistant to Cefotaxine and 63.6% resistant to Augmentin and Cefuroxine while the Gram-negative bacteria were 100% resistant to Nalidixic acid, 80% resistant to Ampiclox, 60% resistant to Augmentin, Imipenem/Cilastatin, Cefuroxine and Nitrofurantoin. Out of the twenty-seven (27) isolates, 74% had a multidrug resistance index ≥ 0.2, 14.9% had ≥ 0.5, and 11.1% had <0.2 Multidrug Resistance Index value. This study revealed that azithromycin, ciprofloxacin, levofloxacin, imipenem/cilastatin, and ofloxacin can be used as a drug of choice for treatment of Gram-positive bacteria associated with hospital wastes while levofloxacin, ofloxacin, ceftriaxone sulbactam, gentamycin, and cefixime could be used for Gram-negative bacteria associated with hospital wastes.

Multidrug-resistance and extended-spectrum beta-lactamase-producing lactose-fermenting enterobacteriaceae in the human-dairy interface in northwest Ethiopia.

Antimicrobial resistance (AMR) is among the top public health concerns in the globe. Estimating the prevalence of multidrug resistance (MDR), MDR index (MDR-I) and extended-spectrum beta-lactamase (ESBL)-producing lactose fermenting Enterobacteriaceae (LFE) is important in designing strategies to combat AMR. Thus, this study was designed to determine the status of MDR, MDR-I and ESBL-producing LFE isolated from the human-dairy interface in the northwestern part of Ethiopia, where such information is lacking. A cross-sectional study was conducted from June 2022 to August 2023 by analyzing 362 samples consisting of raw pooled milk (58), milk container swabs (58), milker's hand swabs (58), farm sewage (57), milker's stool (47), and cow's feces (84). The samples were analyzed using standard bacteriological methods. The antimicrobial susceptibility patterns and ESBL production ability of the LFE isolates were screened using the Kirby-Bauer disk diffusion method, and candidate isolates passing the screening criteria were phenotypically confirmed by using cefotaxime (30 μg) and cefotaxime /clavulanic acid (30 μg/10 μg) combined-disk diffusion test. The isolates were further characterized genotypically using multiplex polymerase chain reaction targeting the three ESBL-encoding- genes namely blaTEM, blaSHV, and blaCTX-M. A total of 375 bacterial isolates were identified and the proportion of MDR and ESBL-producing bacterial isolates were 70.7 and 21.3%, respectively. The MDR-I varied from 0.0 to 0.81 with an average of 0.30. The ESBL production was detected in all sample types. Genotypically, the majority of the isolates (97.5%), which were positive on the phenotypic test, were carrying one or more of the three genes. A high proportion of the bacterial isolates were MDR; had high MDR-I and were positive for ESBL production. The findings provide evidence that the human-dairy interface is one of the important reservoirs of AMR traits. Therefore, the implementation of AMR mitigation strategies is highly needed in the area.

Carvacrol-loaded invasomes efficacy against multidrug resistant isolates of Enterobacteriaceae and housefly

The current study aimed to evaluate the antimicrobial activity of carvacrol-loaded invasomes (CLI) against multidrug-resistant (MDR) Enterobacteriaceae and its mechanical vector, the housefly. CLI were prepared and characterized in the laboratory. Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serovar Enteritidis, Salmonella enterica subsp. enterica serovar Typhimurium, and Klebsiella oxytoca were among the MDR enterobacteriaceae stains investigated. These strains were first isolated and identified from naturally infected chickens. The antibacterial activity of CLI against the MDR isolates was evaluated using the diffusion method. In addition, the insecticidal activity of CLI against housefly larvae and pupae was tested. The MDR index of all evaluated isolates was greater than 20%, indicating that they were all multidrug-resistant. CLI decreased the growth of all isolates except S. Typhimurium and P. aeruginosa at a dose of 0.0125%; however, pure carvacrol inhibited the growth of only Klebsiella oxytoca. Furthermore, both CLI and pure carvacrol inhibited Klebsiella oxytoca growth at different concentrations. CLI inhibited E. coli and S. enteritidis at lower concentrations than pure carvacrol, even at a doubled concentration. Carvacrol and CLI caused significant larval mortality even at low concentrations, with LC50 reached at concentartions of 2.54 and 2.19 µl/ml, respectively. Furthermore, at a low concentration of 3.125 µl/ml, both elicited a high percentage inhibition rate (PIR) in pupae. In conclusion, CLI demonstrated substantial antibacterial action, particularly against MDR isolates, as well as pesticide activity against houseflies.

Phylogenetic Characterization of Resistant Salmonella Strains in Typhoid Fever Patients in Nigeria.

Salmonella species are Enterobacteriaceae associated with typhoid fever. In this study, the distribution of broad-spectrum β-lactamase regulatory genes and genetic relatedness of isolates was determined. Stool samples (400) were collected from patients with fever in Dalhatu Araf Specialist Hospital (DASH), Lafia, Nigeria, between March 2020 and April 2021. Salmonella species were isolated and extended-spectrum β-lactamase distribution was determined among resistant isolates using polymerase chain reaction (PCR). Genetic relatedness of Salmonella species resistant to the 10 first-line antibiotics administered was determined among S typhi isolated. Of the 60 isolates that were confirmed to belong to the genus Salmonella, 12 (20.0%) isolates with bla SHV genes were the most prevalent, blaOXA-1 and blaCTX-M-9 were present in 5 isolates each, while blaCTX-M-4 and blaTEM genes with a prevalence of 1.7% each were the least obtained in the isolates. Two isolates had a multidrug-resistant index (MDRI) of 1, and 2 others were positive with the S typhi staG gene. Sequencing to determine their diversity showed that isolates ST36 and ST138, respectively, had MDRI = 1 and are clustered in a group with a similarity coefficient of 0.00634. The 2 isolates had the highest genetic similarity, which indicates that the genetic diversity between the isolates is low, while Salmonella strain ST313L2 had a high level of genetic distance from the other isolates. The most resistant isolates are closely related which calls for concern.

RESULTS OF BACTERIOLOGICAL MONITORING OF URINARY TRACT INFECTIONS IN CHILDREN OF ZAPORIZHZHIA REGION IN 2023

The aim of the study. To investigate the structure of pathogens of acute urinary tract infections in children of Zaporizhzhia region in 2023 and their sensitivity to antibacterial agents. Materials and methods. The analysis of the structure of pathogens in acute urinary tract infections and the determination of their sensitivity to antibiotics based on the results of bacteriological urine cultures of 322 children with acute urinary tract infections treated at the Zaporizhzhia Regional Children's Hospital in 2023 was performed. For each isolate, the multidrug resistance index was calculated, which allowed to judge the relative integral resistance of the isolated microorganisms to antibacterial drugs. Results. It was established that the most common cause of urinary tract infections in children were gram-negative microorganisms (61.0%), including Escherichia coli (37.4%) and Klebsiella pneumoniae (10.8%) which were most often verified. Gram-positive bacteria were identified in 38.5% of patients, among which the Enterococcus bacteria (29.2%) predominated. High resistance of Escherichia coli to amoxicillin/clavulanate (31.5%), ciprofloxacin (30.1%), cefepime (20.1%), and ceftazidine (20.5%) was detected, including 29 (39.7%) multidrug-resistant isolates of Escherichia coli. The highest resistance of Klebsiella pneumoniae was found to amoxicillin/clavulanate (76.2%), cefepime (33.3%), ceftazidine (33.3%), ciprofloxacin (42.8%), and phosphamycin (28.6%). Multidrug resistance was shown by 76.2% of Klebsiella pneumoniae isolates. The highest susceptibility of Enterococcus bacteria was found to ampicillin (98.2%), linezolid (98.2%), vancomycin (100%), nitrofurantoin (100%), and tigecycline (100%). In no case microorganisms showed complete sensitivity to imipenem, and 17.9% of Enterococcus isolates were resistant to this antibiotic. Conclusions: At the present stage, the dominance of gram-negative microflora among the pathogens of urinary system infections in children remains, among which Escherichia coli (37.4%) continues to be the leading pathogen. The growing role of Enterococcus faecalis (29.2%) as an etiologic factor of urinary tract infections in children was noted, which should be taken into account during the empirical use of antibiotics, especially if there is no effect within the first two days after the start of therapy.

Evaluation of Antibiotic Resistant Bacteria from Organic Fertilized Farm Soils and Waterleaf (Talinum triangulare) in Aluu, Rivers State, Nigeria

The use of antibiotics in agriculture is believed to contribute to the spread of antibiotic resistance, and consumption of vegetables represents a route of direct human exposure to resistant bacteria found in soil. This study was carried out to isolate and characterize antibiotic resistant bacteria from poultry dropping fertilized farm soils in Aluu community Rivers State. Thirty-six (36) soil and waterleaf (Talinum triangulare) samples were collected for a period of three months from the vegetable farms and subjected to standard microbiological procedures such as standard plate counts, identification, sensitivity testing using Kirby-Bauer disk diffusion method and molecular identification. The total heterotrophic bacterial (THB), Staphylococcal, and total coliform counts ranged from2.72±0.52x106 to 7.30±3.54x106Cfu/g; 0x105to 4.30±0.28x105Cfu/g; 0x105 to 21.40±2.26x105Cfu/g and 2.65±0.21x105 to 3.10±0.85x105Cfu/g for FarmsA, B and C respectively. There was a significant difference (p˂0.05) in the total heterotrophic, coliform bacterial count but no significant difference in Staphylococcal and Salmonella-Shigella count between the different vegetable farms sampled in month one. THB, Staphylococcal, Faecal coliform (FC), coliform, Salmonella-Shigella counts ranged from3.80±1.83x106 to 4.69±1.69x106cfu/g; 1.08±0.04x105 to 1.40±0.11x105Cfu/g; 2.00±0.42x104 to 8.30±2.12x104Cfu/g; 3.95±0.21x105 to 5.95±0.21x105Cfu/g and 1.20±0.28x104to 2.40±0.85x104Cfu/g for FarmsA and B respectively. There was no significant difference (p˂0.05) in the THB, Salmonella-Shigella, Total coliform (TC) counts, but there was a significant difference in the Staphylococcal and faecal coliform counts between the different vegetable farms sampled for month two. The bacterial isolates identified were as follows; Escherichia coli, Enterobacter spp, Shigella spp, Salmonella spp, Pseudomonas spp Proteus spp, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis, Serratia marcesens and Serratia nematodiphila. Forty-six (46) bacterial isolates were isolated from the soils and vegetables. Escherichia coli had the highest occurrence (75%) and Bacillus spp (9.09%) had the least occurrence. The susceptibility pattern of Bacillus spp, Staphylococcus spp, Enterobacter spp, Salmonella spp, Escherichia coli, Shigella spp, Pseudomonas sppand Proteus sppwere all susceptible to Ofloxacin, and gentamicin (100%) and resistant to Ceftazidime, Cefixime and Augmentin (100%). Hundred-percent (100%) of the bacterial isolates had multidrug resistance index greater than 0.2 and QnrA resistant gene were found in the resistant bacterial isolates. In conclusion, the use of antibiotic indiscriminately for agricultural purposes should be discouraged because of multiple antibiotic resistance.

Poultry slaughterhouse wastewater as reservoirs for spreading extended-spectrum beta-lactamase-producing Escherichia coli in Abakaliki, Nigeria

Abstract. Ugbo EN, Jacob JI, Effendi MH, Witaningrum AM, Agumah BN, Ugbo AI, Moses BI. 2023. Poultry slaughterhouse wastewater as reservoirs for spreading extended-spectrum beta-lactamase-producing Escherichia coli in Abakaliki, Nigeria. Biodiversitas 24: 4960-4966. The emergence of antimicrobial and multi-resistance extended-spectrum beta-lactamase-producing organisms (ESBL) from the environment and hospitals is a serious global health concern. This research focused on the role of poultry slaughterhouse wastewater as a reservoir for spreading extended-spectrum beta-lactamase-producing Escherichia coli. Exactly 110 poultry slaughterhouses wastewater was collected using sterile universal containers from abattoirs in Abakaliki. The samples were analyzed microbiological for the presence of E. coli using standard techniques. The E. coli isolates found were subjected to a sensitivity test to antibiotics using the Kirby-Bauer disc diffusion method and further tested for ESBL-producing E. coli using a double disc synergy test and chromogenic media Brilliance ESBL agar. Of 110 wastewater samples analyzed, 55 (50.0%) were positive for E. coli contamination. The E. coli isolates showed antibiotic resistance ranging between 21.8% (amoxicillin clavulanic acid) to 69.1% (tetracycline), and good susceptibility was observed on cefepime (72.7%). Thus, among 55 E. coli isolates, 11(20.0%) were ESBL-producing E. coli with resistance patterns as follows: TET-STX-CAZ-CTX-FEP (four isolates); TET-STX-CAZ-CTX (four isolates); TET-STX-CAZ-CTX-FEP-CRO (three isolates), and average multidrug resistance index of 0.7. This study revealed that poultry slaughterhouse wastewater was a reservoir for multidrug resistance ESBL-producing E. coli. Therefore, properly treating wastewater from poultry production or farms before disposing into the drainage channel or water bodies is highly recommended to avoid spreading ESBL-producing organisms from animal waste products to humans and their environment.

Prevalence and Antimicrobial Susceptibility Pattern of Salmonella among Food and Food Vendors in Port Harcourt, Nigeria

Salmonella contamination in ready-to-eat food is seen as a health risk, and improper food processing puts customers at risk. This study therefore was carried out to investigate the prevalence and antimicrobial susceptibility pattern of Salmonella isolated from food vendor’s hands and work bench in Port Harcourt, Rivers State. Ninety (90) samples were collected for a period of three months from three different location in Port Harcourt with five sampling points in each locations. Samples were analyzed for the presence of Salmonella organism using standard microbiological procedure for enumeration and identification. Salmonella-Shigella (SSC) counts ranged from 1.9+4.9x103cfu/g to 3.8+1.4x103cfu/g for choba and Aggrey road respectively in cooked rice, 0.2+0.3x103cfu/g to 0.9+0.4x103cfu/g for Mile 3 and Aggrey road respectively in Moi Moi, 0.6+0.5x103cfu/g to 1.5+1.5x103cfu/g for Aggrey road and Mile 3 in Salad, and 1.6+0.7x103cfu/ml to 2.6+1.7x103cfu/ml for Choba and Aggrey road in Egusi soup. Six (6) Salmonella spp were isolated in Salad and workbench from Mile 3 and Choba respectively. Salmonella showed a decreasing trend of resistance in the order: Ceporex, Gentamycin and Augmentin (100%) > Nalidixic acid (83.3%) > Septrin and Streptomycin (66.7%) > Tarivid, Pefloxacin and Ampicilin (33.3%). The molecular identification of the Six (6) isolates using polymerase chain reaction confirmed 100% Salmonella spp isolates. The six (6) Salmonella isolates were identified with blaTEM and blaSHV gene 6(100%). Out of the Six (6) isolates, 100% had multidrug resistance index ≥ 0.2 while 0.00% had <0.2 as their Multidrug Resistance Index. Conclusively, this study revealed prevalence of Salmonella spp in food and food vendors in Rivers State, posing a serious threat to consumers. Indiscriminate use of antibiotics should be discouraged to reduce the prevalence of resistant strains of Salmonella.

Pseudomonas isolates from raw milk with high level proteolytic activity display reduced carbon substrate utilization and higher levels of antibiotic resistance

Milk is an ideal food but can be easily contaminated during milking, transportation and storage by Pseudomonas spp. That secrete proteases resulting in milk spoilage. However, genomic features and functional metabolic diversity of Pseudomonas with different protease activity have not been extensively studied. With this aim, the genomic features and functional metabolic diversity of Pseudomonas with differing proteolytic activity were evaluated using whole genome sequencing and metabolomic approaches, respectively as well as, the presence of genes encoding the aprX protease and antibiotic resistance were screened. Pseudomonas strains with higher protease activity possessed more carbohydrate transport and metabolism, signal transduction, secondary metabolite synthesis, polysaccharide lyase and carbohydrate esterase genes. These isolates were more efficient at carbon source utilization and this phenotype was positively related to incubation temperature. The high protease activity group also displayed a multidrug resistance index that was significantly higher than that for the low protease activity group. This is the first report comparing the gene function and metabolic diversity of Pseudomonas with differing levels of proteolytic activity in raw milk.

Antibiogram of Staphylococcus Species Isolated from Some Abattoirs in Rivers State, Nigeria

Foodborne contamination especially by Staphylococcus species is of concern as many isolates of the genus have been reported for their multidrugs resistance. The study was aimed at determining the antimicrobial resistance of Staphylococcus species isolated from abattoir. A total of 100 samples were collected from five different surfaces; bucket swab, butchers hand swab, table swab, knife swab and meat swab from four abattoirs in of Rivers State, Nigeria. The samples were transported aseptically to laboratory for immediate analysis. The samples were cultured on mannitol agar (MSA) using standard microbiology technique to isolate Staphylococcus species. Hemolysis test was carried out and susceptibilities of isolates against a panel of 10 antibiotics were determined using the Kirby-Bauer disk diffusion method, and the multiple antibiotic resistance (MAR) index of the isolates were determined. Out of the 43 isolates of Staphylococcus from the abattoirs, 51.1% were coagulase positive while 48.9% were coagulase negative. 18.6% of the Staphylococcus species produced beta hemolysis and 16.28% showed alpha hemolytic activity. Staphylococcus species produced resistance to amoxicillin (88.4%), zinnacef (88.4%), ampiclox (88.4%), receptrin (60.5%), streptomycin (37.2%), septrin (37.2%), erythromycin (34.5%), and gentamicin (39.5%), ciprofloxacin (20.9%) and perfloxacin (16.8%). The multidrug resistance index (MAR index) showed that 2.35% of the isolates showed complete resistance to all the 10 antibiotics tested with MAR index of 1, 4.6% recorded MAR index of 0.1, 4.6% recorded MAR index of 0.2. Majority of the isolates of Staphylococcus species associated with meat and materials or equipment (such as bucket, knife) used in abattoir and the hands of butcher are multidrug resistance and is of great medical concern hence there is need to regulate the indiscriminate use of antibiotics.

Prevalence of Salmonella in Sea Foods and Its Resistance to Drugs

Salmonella are non-spore forming, predominantly motile enterobacteria with peritrichous flagella (all around the cell body). They are intracellular pathogens causing illness. They usually invade only the gastrointestinal tract and cause Salmonellosis. Humans become infected most frequently through contaminated food or water. Sea foods are mostly associated with Salmonella infections. This study therefore was carried out to isolate Salmonella and determine the antibiotic sensitivity patterns of the Salmonella isolates from sea foods associated with food-borne diseases. These sea foods were obtained from three different selected markets within Port Harcourt Metropolis, Rivers State. A total of three (3) sea food samples were collected at random from each of the market. The samples were put through standard microbiological techniques. The result of total heterotrophic bacterial counts showed that Prawn from Mile 1 market had high count of 2.06x108 cfu/g while Crab from Creek Road market had the least count of 1.15x108 cfu/g. The result of total coliform count showed that Crab from Mile 1 market had the highest count of 2.86x106 cfu/g while Periwinkle from Creek Road market had the least count of 2.30x106 cfu/g. Also, the result of salmonella shigella count showed that Periwinkle from Mile 1 market had the least count of 0.76x105 cfu/g while Crab from Creek Road market had the highest count of 2.80x105 cfu/g. Characteristics of bacterial isolates from the sea foods showed that Proteus sp., Bacillus spp., Pseudomonas sp., Micrococcus sp., Staphylococcus aureus, Escherichia coli, and Salmonella spp. were present and identified from the samples. The result of the antibiotic susceptibility pattern of the Salmonella isolates showed that all the isolates were 100% susceptible to Tarivid, Peflacine, Septrine and Ciprofloxacin while they exhibited 100% intermediate sensitivity or resistance to the other antibiotics such as Ceporex, Nalidixic acid, Ampicillin, Gentamycin, Streptomycin, and Augmentin. The result of the multidrug resistance index of 3 (100%) of the 3 Salmonella isolates had a MAR index ≥ 0.2. The presence of these organisms in sea food could pose threat of food-borne infection as well as resistance to some commonly used antibiotics. Enforcement of existing laws associated with food storage, preparation and hygiene should be done in order to prevent or reduce these food-borne diseases such as salmonellosis.

Evaluation of the Antibacterial Activity of Extracts of Betula Pendula (Silver Birch) Bark Against Uropathogenic Microorganisms

The aim of this work was to evaluate the antibacterial activity of the aqueous and ethanolic extracts of Betula pendula (silver birch) bark against various microorganisms causing urinary tract infections.Material and methods. Aqueous and ethanol extracts of biologically active compounds from Betula pendula bark were tested against ten clinical uropathogenic strains (gram-positive bacteria — Kocuria rhizophila 1542, Staphylococcus simulans 5882, Enterococcus avium 1669, Enterococcus faecalis 5960, and Corynebacterium spp. 1638; gram-negative bacteria — Enterococcus cloacae, Morganella morganii 6392, Escherichia coli 1449, Seratia mansescens 6441, and Achromobacter xylosoxidans 4892). Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 25922 were used as reference standards for Gram-positive and Gram-negative microorganisms. The sensitivity of the experimental strains to the antibiotic was evaluated by the Kirby–Bauer disk diffusion method, while the antibacterial activity of the obtained extracts was evaluated using the agar diffusion method. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were also determined via the serial dilution method.Results. The bark of B.pendula contained 78.85% of dry matter. The volume yield of the aqueous (AE) and ethanol extract (EE) was 74.66% and 86.66% (v/v), respectively, while their mass yields were 6.59% and 10.65% (w/w). K.rizophilia 1542 and Corynebacterium spp. 1638 were the most resistant bacteria with a multidrug resistance index of 0.45. AE and EE were active against all microorganisms tested. MIC of AE ranged from 8 to 32 mg/ml, while MIC of EE ranged from 2 to 16 mg/ml.Conclusion. An aqueous extract of Betula pendula bark exhibits weak antimicrobial activity, while the ethanol extract exhibits a more pronounced antimicrobial activity, but has a bacteriostatic effect.

Bacteriological Quality of Money from the Cash Registers of Banks and Markets in the Port Harcourt Metropolis and Bacterial Susceptibility to Antibiotics

Bacterial contamination of money acts as vehicle for the transmission of pathogenic and drug resistant organisms. This study was carried out to investigate the bacteriological quality and antibiotic susceptibility patterns of money bank cash-points and markets in Port Harcourt metropolis. Two hundred and eighty-eight (288) naira notes belonging to the following denominations, N5, N10, N20, N50, N100, N200, N500 and N1000 were collected for three months from banks and markets and subjected to standard microbiological procedures such as standard plate counts, identification, sensitivity testing using Kirby-Bauer disk diffusion method. Total heterotrophic bacterial count (THB) ranged from 0.6±0.00x106 to 12.80±9.19x106cfu/g; 0.65±0.21x106 to 13.05±9.55x106cfu/g; 0.65±0.20x106 to 8.05±2.48x106Cfu/g in N5 and N100 from Access Bank (AB), United Bank of Africa (UBA) and Guaranty Trust Bank (GTB), respectively. There was a significant difference (p≤0.05) in the THB between the different naira notes. Total coliform count (TCC) ranged from 0.00 x104 to 56.90±28.43x104Cfu/g; 0.00 x104 to 61.90±35.49x104Cfu/g; 0.00x104 to 17.75±12.79x104Cfu/g in N5, and N100 for AB, UBA and GTB respectively. THB ranged from 2.69±1.55x106 to 9.95±3.22x106cfu/g; 3.00±1.69x106 to 12.30±9.89x106cfu/g; 3.30±1.82x106 to 17.30±6.97x106cfu/g in N1000, N10 and N100 naira for Mile 1, Mile 3 and Creek road markets respectively. There was a significant difference (p≤0.05) in the THB between the different naira notes sampled in both Banks and markets. TCC ranged from 0.77±0.28x104 to 45.59±10.18x104Cfu/g; 0.78±0.88x104 to 40.59±3.11x104Cfu/g; 1.45±0.07x104 to 55.60±10.18x104Cfu/g in N1000 and N100 Mile 1, Mile 3 and Creek road market respectively. Faecal coliform count ranged from 0.00x103 to 1.53±0.15x103Cfu/g for only mile 1 market. There was a significant difference (p≤0.05) in the total and faecal coliform counts between the different naira notes sampled in Banks and markets. The bacterial isolates identified were; Escherichia coli, Staphylococcus aureus, Micrococcus luteus, Bacillus cereus and Serratia marcesens. Twenty-one (21) bacteria were isolated from Banks and Markets. Bacillus cereus, Staphylococcus aureus, Micrococcus leteus, Escherichia coli and Serratia marcesens were significantly high in N100 (41.67%) (66.67%), (100%), (100%) respectively. Staphylococcus spp was susceptible to Gentamicin (33.3%), Bacillus spp to Ofloxacin (100%), Micrococcus spp to Ofloxacin and Gentamicin (100%), Serratia spp and Escherichia coli were susceptible to Ofloxacin, Gentamicin and Nitrofurantoin (100%) and they were all resistant to Ceftazidime, Cefixime, Cloxacillin, Augmentin, Cefuroxime and Ceftriaxone (100%)> Erythromycin and Gentamicin (75%). Hundred-percent (100%) of the bacterial isolates had multidrug resistance index greater than 0.2. Public awareness on personal hygiene should be encouraged while drug abuse should be discouraged.

Molecular Characterization and Antimicrobial Resistance of Pathogenic Escherichia coli Strains in Children from Wolaita Sodo, Southern Ethiopia.

Introduction Pathogenic Escherichia coli strains cause diarrheal infection in children due to their virulence factors. A nonanalytical observational study followed by a purposive sampling technique was conducted from October 2017 to June 2018, to determine the antimicrobial susceptibility patterns and molecularly detect pathogenic Escherichia coli strains in under-five children at Wolaita Sodo town using molecular and the Kirby-Bauer disc diffusion method. Result In the current investigation, out of 110 stool samples, Escherichia coli was isolated in 68 (61.8%) (95% CI: 52.1–70.9%). Out of 68 Escherichia coli isolates, 61.9% of Escherichia coli isolates were resistant, 9.4% were intermediately resistant, and 28.7% were susceptible. Among the antimicrobial agents, 91.2% of Escherichia coli isolates were highly sensitive to ciprofloxacin followed by norfloxacin (86.8%). Virulence genes were detected in 55.9% (38/68) (95% CI 52.1%–70.9%) of isolates. The following genes were detected: stx1 (6 (8.8%)), stx2 (4 (5.9%)), eaeA (5 (7.4%)), eaat (14 (20.5%)), and St (4 (5.9%)). Out of 68 Escherichia coli isolates, 43 (63.2%) isolates had shown multidrug resistance patterns. Furthermore, 11.8% of Escherichia coli isolates had shown resistance to eight different drugs. The multidrug resistance index value of Diarrheagenic Escherichia coli pathotypes was greater than or equal to 0.4, which indicates the high risk of resistance. Conclusion This study demonstrated important pathogenic Escherichia coli strains and multidrug resistance in isolates containing virulence genes. Wise use of antimicrobials and improving the hygienic practices amongst parents of children reduce its occurrence. Therefore, appropriate usage of antimicrobial agents should also be highly practiced in hospitals.

Assessment of Antimicrobial Activity of Ethanolic and Aqueous Extracts of Aesculus hippocastanum L. (Horse Chestnut) Bark against Bacteria Isolated from Urine of Patients Diagnosed Positive to Urinary Tract Infections

The search for new antimicrobials is essential to address the worldwide issue of antibiotic resistance. The present work aimed at assessing the antimicrobial activity of Aesculus hippocastanum L. (horse chestnut) bark against bacteria involved in urinary tract infections (UTIs). Bioactive compounds were extracted from A. hippocastanum bark using water and ethanol as solvents. The extracts were tested against 10 clinical uropathogenic strains including five Gram-positive and five Gram-negative bacteria. Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 25922 were used as reference bacteria. The susceptibility to antibiotics was assessed using the Kirby Bauer disc diffusion method and the antibacterial activity of the extracts was evaluated using the well diffusion method. The Minimum inhibitory concentration (MIC) and the minimum bactericidal concentrations (MBC) were asseded by the microdilution method. A. hippocastanum bark possessed a dry matter content of 65.73%. The aqueous extract (AE) and ethanolic extract (EE) showed a volume yield of 77.77% and 74.07% (v/v), and a mass yields of 13.4% and 24.3% (w/w) respectively. All the bacteria were susceptible to amoxiclav, imipenem and ceftriaxone but the clinical strains were resistant to at least one antibiotic. Kocuria rizophilia 1542 and Corynebacterium spp 1638 were the most resistant bacteria both with multidrug resistance index of 0.45. Except AE on Proteus Mirabilis 1543 and Enterococcus faecalis 5960 (0 mm), both AE and EE were active against all the microorganisms tested with inhibition diameters (mm) which ranged from 5.5-10.0 for AE and 8.0-14.5 for EE. The MICs of EEs varied from 1-4 mg/mL while those of AEs varied from 4-16 mg/mL. The ethanolic extracts (EE) were overall more active than the aqueous ones. The A. hippocastanum bark extracts had overall weak antibacterial activity (MIC ≥0.625 mg/mL) and bacteriostatic potential (MBC/MIC ≥16) on both Gram-positive and Gram-negative bacteria.

Multidrug-resistant and enterotoxigenic methicillin-resistant Staphylococcus aureus isolated from raw milk of cows at small-scale production units.

Objective:Staphylococcus aureus (S. aureus) has evolved as one of the most significant bacteria causing food poisoning outbreaks worldwide. This study was carried out to investigate the prevalence, antibiotic sensitivity, virulence, and enterotoxin production of S. aureus in raw milk of cow from small-scale production units and house-raised animals in Damietta governorate, Egypt.Material and Methods:The samples were examined bacteriologically, and antimicrobial sensitivity testing was carried out. Moreover, isolates were characterized by the molecular detection of antimicrobial resistance, virulence, and enterotoxin genes.Results:Out of 300 milk samples examined, S. aureus was isolated from 50 samples (16.7%). Antibiotic sensitivity testing revealed that isolates were resistant to β-lactams (32%), tetracycline (16%), and norfloxacin (16%); however, they showed considerable sensitivity to ceftaroline and amikacin (72%). Multidrug-resistance (MDR) has been observed in eight isolates (16%), with a MDR index (0.5) in all of them. Of the total S. aureus isolates obtained, methicillin-resistant S. aureus (MRSA) has been confirmed molecularly in 16/50 (32%) and was found to carry mecA and coa genes, while virulence genes; hlg (11/16, 68.75%) and tsst (6/16, 37.5%) were amplified at a lower percentage, and they showed a significant moderate negative correlation (r = −0.59, p-value > 0.05). Antibiotic resistance genes have been detected in resistant isolates relevant to their phenotypic resistance: blaZ (100%), tetK (50%), and norA (50%). Fifty percent of MRSA isolates carried the seb enterotoxin gene.Conclusion:High detection rate of MRSA and MDR isolates from milk necessitates the prompt implementation of efficient antimicrobial stewardship guidelines, especially at neglected small-scale production units.

The Antimicrobial Pattern of E. coli Isolated from Various Animal Sources at El Behera Governorate

The emergence of multidrug resistant E. coli isolated from animal and animal products become the main wary of food safety and public health authorities worldwide. The present study was undertaken to recognize the most important serovars and the antimicrobial sensitivity patterns of E. coli isolated from animals and animal products. Results showed that out of the 300 examined samples (75 cows mastitic milk, 77 buffalo's mastitic milk, 40 cattle meat, 43 buffaloes meat and 65 chicken meat), 60 isolates (20 %) were confirmed as E. coli based on cultural characteristic and biochemical identification. These isolates were further identified serologically, the obtained isolates were categorized as O26:H11 (4/22, 18%), O91:H21 (3/22, 13.6%), O111:H2 (2/22, 9%), O146:H21 (2/22, 9%), O1:H7 (1/22, 4.5%), O55:H7 (1/22, 4.5%), O119:H6 (1/22, 4.5%), O121:H7 (1/22, 4.5%), O78 (2/22, 9%), O125: H21 (2/22, 9%), O128 : H2 (1/22, 4.5%), O159 (1/22, 4.5%), and O44:H18 (1/22, 4.5%). The isolates were further tested for antimicrobial resistance against 14 commercial antibiotics. The obtained results showed that the isolates were resistant to Erythromycin (22/22, 100٪), Streptomycin (21/22, 95.5٪), Nalidixic acid (17/22, 80٪), Clindamycin (14/22, 63.6٪) and Sulphamethozole (11/22, 50٪). Lower resistance rate was observed in case of Cephalothin (9/22, 40.9٪), Tetracycline (7/22, 31.8٪), Ampicillin (6/22, 27.3٪), Colistin (5/22, 22.7٪), Ciprofloxacin (5/22, 22.7٪), Amikacin (4/22, 18.2٪), Levofloxacin (2/22, 9.1٪), Gentamycin (2/22, 9.1٪) and Meropenem was (1/22, 4.5٪). In conclusion, there a wide variation in E. coli serogroups isolated from different animals and animal products. These serogroups exhibited a high multidrug resistance index against the commonly used antibiotics. Therefore, regular monitoring of the potential sources of human infection with E. coli will help establish accurate preventive and curative measures.

Antibiotic resistance pattern of uropathogenic Escherichia coli isolated from children with symptomatic urinary tract infection in Moscow, Russia

Background and Aim: Uropathogenic Escherichia coli (UPEC) is commonly involved in urinary tract infections (UTIs), which are generally treated with antibiotics. However, the emergence of multidrug-resistant (MDR) strains of UPEC has made the treatment difficult. There is thus a need to continuously assess their sensitivity to antibiotics. This study aimed to determine the antibiotic resistance patterns and MDR phenotypes of UPEC strains isolated from children diagnosed with UTIs at the Russian Children's Clinical Hospital in Moscow, Russia. Materials and Methods: Kirby–Bauer's disc diffusion method was used to study the sensitivity to antibiotics of 106 UPEC isolates from urine specimens from children (aged from 9 months to 18 years old) diagnosed with UTIs. The results were interpreted in accordance with the Clinical and Laboratory Standards Institute guidelines and the correlations of variables with the degree to which each antibiotic inhibited the UPEC strains in terms of diameter on the disc were determined using Spearman's rank correlation test. A t-test and principal component analysis were performed to visualize the correlations of the susceptibility of UPEC to antibiotics with the age and sex of the patients. Statistical significance was set at p≤0.05. Results: Among the 106 UPEC strains tested, none (0%) showed resistance to fosfomycin (FO), while 84 (79.2%) were resistant (R) to at least one antibiotic. The highest rates of resistance were observed to amoxicillin (69.8%), ampicillin (62.3%), cefazolin (39.6%), trimethoprim (TR) (37.7%), ceftriaxone (34.9%), and tetracycline (33.0%). Interestingly, 22 (20.8%) strains were R to imipenem. UPEC isolates from males aged 1-6 years were more R to antibiotics than those from the other groups, with the exception of TR, to which UPEC isolates from females aged 13-18 years old were less sensitive (S). The multidrug-resistance (MDR) index ranged between 0.00 and 0.75 and we found that more than a quarter of UPEC (31/106) had an MDR index ≥0.5 and only 22 (20.7%) strains were S to all antibiotics tested (MDR index=0). Finally, Spearman's rank correlation test showed that, with the exception of FO, there were correlations between the inhibition diameters of all other antibiotics. Conclusion: FO is the only antibiotic to which all UPECs were S and may be suggested as the first line of treatment for UPEC. Further research is needed to continue monitoring antibiotic resistance and to investigate the genetic features associated with such resistance observed in this study.

Multidrug resistance, biofilm formation, and virulence genes of Escherichia coli from backyard poultry farms.

Background and Aim:Backyard chicken flocks have traditionally been regarded as an essential food source in developed countries; however, they may act as reservoirs and spread various zoonotic bacterial pathogens. This study was designed to investigate the prevalence, phenotypic resistance, biofilm formation (BF), and pathotypes of Escherichia coli isolates from backyard poultry farms.Materials and Methods:Cloacal swabs (n=150) and internal organs (n=150) were collected from 30 backyard chicken flocks; 20 of them were experiencing systemic infection, and the other ten were apparently healthy. Samples were bacteriologically examined for E. coli isolation. Isolates were identified biochemically by the VITEK® 2 COMPACT system (BioMérieux, France). For molecular identification, 16S rRNA was amplified and sequenced. Ten antimicrobials were selected for E. coli antimicrobial susceptibility testing. The minimum inhibitory concentration for each antimicrobial was determined. The extended-spectrum β-lactamase activity in isolates was investigated using cephalosporin/clavulanate combination disks. The ability of isolates for BF was determined by the microtiter plate method. Thirteen virulence genes linked to different E. coli pathotypes and two serotype-related genes were investigated by real-time polymerase chain reaction.Results:Eighty-six E. coli strains were isolated from 30 backyard chicken flocks. The isolates were biochemically identified to the species level. Genetically, sequences of the 16S rRNA gene showed >98% identity with E. coli in the National Center for Biological Information database. The frequency of isolation from diseased flocks was significantly higher (p<0.05) than apparently healthy flocks; 63.9% of the isolates were recovered from cloacal swabs and 36.04% were recovered from internal organs. E. coli isolates showed high resistance to ampicillin (AMP; 75.6%), gentamicin (39.5%), and tetracycline (29.1%). However, none of the isolates were resistant to imipenem. A variable drug resistance profile for E. coli isolates was reported. Twenty-one (24.4%) isolates were sensitive to all ten antimicrobials. Seven (8.1%) isolates were resistant only to AMP, and 28 (32.6%) were resistant to two antimicrobials, whereas the remaining 30 (34.9%) isolates showed multidrug resistance (MDR). Of the 86 isolates, 8 (9.3%) were confirmed as extended-spectrum β-lactamase (ESBL)-producing E. coli by the combination disk diffusion method. All ESBL isolates were MDR with an MDR index of 0.5-0.6. Fifty-seven (66.3%) isolates were capable of forming biofilms; 22 (25.6%) of them were strong biofilm producers, 24 (27.9%) moderate producers, and 11 (12.8%) weak producers. A statistically significant pairwise correlation was obtained for MDR versus BF (r=0.512) and MDR index versus BF (r=0.556). Based on virulence gene profiles, five pathotypes were identified, including enteropathogenic E. coli (39.5%), avian pathogenic E. coli (32.53%), enterohemorrhagic E. coli (EHEC; 9.3%), enterotoxigenic E. coli (ETEC; 5.8%), and enteroaggregative E. coli (EAEC; 1.2%). The lower frequency of EAEC and ETEC was statistically significant than other pathotypes. Three isolates were identified as O157 based on the detection of the rbfO157 gene.Conclusion:This study reported a high prevalence of MDR, suggesting the misuse of antimicrobials in backyard chicken farms. The emergence of ESBL and EHEC isolates in backyard chickens is a public health concern. Furthermore, the backyard flocks environment may harbor different pathogenic bacteria that may enhance the persistence of infection and the transmission to in-contact humans. Regular monitoring for the occurrence of MDR and the zoonotic pathotypes among E. coli in backyard chicken flocks is recommended, as these bacteria can transmit to humans through food products or contaminated environments.

Prevalence of Shigella spp and Methicillin-Resistant Staphylococcus aureus in Select Commercially Processed Meat Products

Aim: The aim of this study was to determine the prevalence of Shigella spp and methicillin resistant Staphylococcus aureus in some selected commercially processed ready-to-eat snail (Archachatina marginata) and edible worm (Rhynchophorus phoenicis).&#x0D; Study Design: Cross-sectional analysis.&#x0D; Place and Duration of Study: Samples were obtained from vendors along the Benin-Sapele express road, South-South Nigeria, over a two months period (November to December, 2019).&#x0D; Methodology: The enumeration of total heterotrophic counts, total Shigella counts and total Staphylococcus aureus counts were done using nutrient agar, Salmonella Shigella agar and mannitol salt agar respectively. All isolates were further identified by their cultural, morphological and biochemical characteristics. Methicillin resistant Staphylococcus aureus were identified using methicillin-oxacillin agar, while the antibiogramic profile of selected isolates and their multidrug resistant profile were done according to the CSLI guideline.&#x0D; Results: The mean heterotrophic bacterial counts (THCs) obtained in Rhynchophorus phoenicis ranged from 0.00 × 10³ ± 0.00 cfu/g to 500.00 × 10³ ± 0.00 cfu/g, while the mean THCs obtained in Archachatina marginata ranged from 13.3 × 10³ ± 1.15 cfu/g to 500 × 10³ ± 0.00 cfu/g. The total Staphylococcus aureus obtained in Rhynchophorus phoenicis was between 0.00 × 10³ ± 0.00 cfu/g to 294 × 10³ ± 4.7 cfu/g and the Shigella bacteria counts from 0.00 × 10³ ± 0.00 cfu/g to 258 × 10³ ± 14.64 cfu/g. The antibiogram of Staphylococcus aureus isolated from both Rhynchophorus phoenicis and Archachatina marginata, found the majority of isolates (96.67%) to be resistant to ceftazidime and cefuroxime, while 83.33% of the isolates were sensitive to ofloxacin. All Staphylococcus isolates were resistant to ceftriaxone, cloxacillin and amoxycillin clavulanate. The multidrug resistant index recorded was between 37.5 and 100. All Staphylococcus (9) isolates tested for methicillin resistance was observed to be positive.&#x0D; Conclusion: This study demonstrated that RTE vended meat products sold in along Benin City By-pass, contains Shigella and methicillin resistant Staphylococcus species, which are potential foodborne pathogens and efforts should be made at eliminating them from these vended meat products.