Multi Antibiotic Resistance Index Research Articles

Evaluation of Multi-antibiotic Resistance Index (MAR) and Molecular Characterization of Pseudomonas aeruginosa Isolated in Pathological Products from Chad

Pseudomonas aeruginosa is a bacterium present in several ecological niches causing opportunistic infections in humans. In developing countries like Chad, this bacteria is responsible for various human pathologies. High-priority pathogens, such as Salmonella and Shigella, represent a particularly heavy burden in low- and middle-income countries, as do Pseudomonas aeruginosa and Staphylococcus aureus, which pose serious problems in health centers. The objective of this study was to evaluate the multi-antibiotic resistance index and to carry out a molecular characterization of Pseudomonas aeruginosa isolated in pathological products from Chad using by rep-PCR (GTG)5. Antibiotic resistance phenotypes were obtained by interpreting the results of our previous study on antimicrobial resistance of Pseudomonas aeruginosa isolated from human infections in N'Djamena, Chad. In this study, two phenotypes were obtained that is XDR and MDR. Thus, 24/36 of strains came from urine and 29/36 strains were MDR. The multi-antibiotic resistance (MAR) indexes varied from 0.33 to 0.86. The rep-PCR allowed good discrimination of Pseudomonas aeruginosa strains in this study. These isolates were grouped based on characteristic bands appearing in their genetic fingerprint profiles. In this study, Pseudomonas aeruginosa strains isolated from urinary tract infections and pus were classified as XDR and MDR type strains.

Phenotypic and genotypic characterization of antimicrobial resistance in Enterococcus species from dairy niches

The present work was carried out to investigate the prevalence of antimicrobial resistance (AMR) in dairy enterococci. Dairy samples comprising of raw milk and traditionally fermented milks were collected from local surrounding areas and a total of 140 enterococci strains were isolated from the dairy samples and further, 104 isolates were tested for antimicrobial susceptibility. The most prevalent resistance was observed against cephalosporins followed by fosfomycin, rifampicin, erythromycin etc. Multi-antibiotic resistance index of the tested isolates ranged from 0.09 to 0.36. Minimum inhibitory concentration testing showed very high cut-offs against erythromycin, tetracycline, cefepime and cefotaxime in certain isolates. Further, Extended Spectrum β-lactamase (ESBL) production tested by double disc synergy method confirmed 9 isolates as ESBL positive along with the presence of blaTEM gene in certain isolates. In addition, AMR genes against aminoglycoside, tetracyclines, macrolides, chloramphenicol along with multi-drug transporter gene (emeA) was also detected in many isolates. The resistant enterococci isolates also showed a high frequency of the presence of mobile genetic elements viz. Transposons (Tn-5397 and Tn-916/Tn-1545 family) and Integrons (intI1, intI3). Finally, virulence traits such as gelatinase and hemolysin production along with gelE and cylA genes was also detected in many of the resistant enterococci isolates. Although, the virulence traits could not be directly correlated with the AMR resistance of the isolates, the results from the study demonstrated the wide prevalence of AMR in dairy enterococci that could greatly contribute to the problem of antimicrobial resistance in the food chain.

Multidrug resistant profiles of clinical isolates of Escherichia coli, Salmonella typhi and Shigella species

In order to determine the antibiotic susceptibility pattern and multi antibiotic resistance index of E.coli, Salmonella typhi and Shigella species, a total of 32 pre-identified clinical isolates were obtained and subjected to conventional biochemical and gram staining reaction for confirmation. Antibiotic susceptibility testing to ten commonly prescribed antibiotics (Amplicin (PN:30 mcg), Gentamicin (GEN:10 mcg), Streptomycin (STR:30 mcg), Septrin (SXT:30 mcg), Nalidixic acid (NAL:30 mcg), Ciprofloxacin (CIP:10 mcg), Augmentin (AU:30 mcg), Reflacin (PEF:10 mcg), Tarivid (OFX:10mcg), and Ceporexin (CEP:10 mcg) was considered using disc agar diffusion method and reported with the Clinical and Laboratory Standard Institute Interpretative chart. Zones of inhibition were measured and Multiple Antibiotic Resistance index was calculated. Salmonella isolates demonstrated a high rate of susceptibility to Tarivid (90%), Reflaxin (90%), Gentamycin (80%), ciprofloxacin (60%), Augmentin (80%) and moderately susceptible to Streptomycin (40%) and Septrin (20%) with high resistance to Nalidixic acid (90%), Amplicin (90%) and Ceporexin (70%). Nearly all Shigella isolates were susceptible to Augmentin (80%), Reflaxin (90%), Tarivid (80%), intermediate susceptibility was also noted in Streptomycin (40%) and Ceporexin (50%) while high resistance to ciprofloxacin (80%), Amplicin (90%), Septrin (70%) and Nalidixi acid (90%). High rate of resistance was observed to nearly five antibiotics in E,coli with low susceptibility rate to all the antibiotics tested. All the isolates had Multiple Antibiotic Resistance index greater than 0.2 and above. This may be as a result of previous exposure to antibiotics and development of resistance to commonly prescribed antibiotics, hence, antimicrobial susceptibility testing is imperative in selecting therapeutic options.

Fecal Carriage of Colibactin-Encoding Escherichia coli Associated With Colorectal Cancer Among a Student Populace.

Fecal carriage of the colibactin (clb) gene in Escherichia coli is described as a source that could promote carcinogenesis, progressing to colorectal cancer. The present study investigated the demographic, dietary, and antibiotic consumption variables as correlates for fecal carriage of clb+/E coli among the student populace. In a randomized cross-sectional survey, E coli (N = 136) from the fecal samples of eligible students were characterized and evaluated for antibiotic resistance, β-lactamase (blm), biofilm, virulence factor production, and strain tryptophan reverse mutagenic activity. The encoded clb+/E coli were analyzed for correlates with principal component analysis. Of all the E coli strains, a low rate of 2 clb+/E coli (1.5%) and higher rates of biofilm (13.2%) and blm producers (11.8%) were recorded among the mutant strains as compared with the nonmutant types. All the clb+/E coli showed complete resistance to amoxicillin, Augmentin (amoxicillin and clavulanate), gentamicin, and trimethoprim/sulfamethoxazole. The fecal clb-encoded E coli (1.5%) were not associated with demographic status, fiber-based food (odds ratio [OR], 1.03; 95% CI, 56.74-138.7; P = .213), alcohol (OR, 1.27; 95% CI, 61.74-147.1; P = .221), antibiotic consumptions (OR, 1.11; 95% CI, 61.29-145.3; P = .222), and handwashing (OR, 1.17; 95% CI, 60.19-145.5; P = .216). The hierarchical cluster of blm+/E coli revealed high-level resistance with a multiantibiotic resistance index ≥0.2 (P < .05). Only 12% of all strains were tryptophan mutant/blm+, and 1.5% of clb+/ECblm+ were observed in fecal samples with a 452-base pair size. Trimethoprim/sulfamethoxazole and biofilm production positively regressed with clb expression (P > .05). Principal component analysis score plot indicated an association of clb+/ECblm+ with dietary pattern, alcohol, blm, and hemolysin production. The combined activity of blm and biofilm production in the gut microbiota could promote clb+/E coli colonization, facilitating genotoxin production and possible colorectal cancer induction.

Microbiota and its antibiotic resistance profile in avocado Guatemalan fruits (Persea nubigena var. guatemalensis) sold at retail markets of Ibarra city, northern Ecuador.

Avocados are typically sold in unsanitary conditions at the retail markets in Ecuador, which can raise the risk of microbial contamination. These microorganisms could exhibit multi-antibiotic resistance (MAR), being a serious threat concern to human health. In this study, we aimed to evaluate the microbiota and its antibiotic resistance profile in avocado Guatemalan fruits (Persea nubigena var. guatemalensis), at ripe stage: immature, firm light green (ready to eat in 4 days), peel (AFPE) and pulp (AFPU), and mature intense green (ready to eat) peel (AMPE) and pulp (AMPU), to gain baseline information on the prevalence of MAR bacteria. Culture-independent (16S rRNA metagenomics) and culture-dependent approach (to detect specific indicator microorganisms) were used. Moreover, antibiotic susceptibility of selected target indicator bacteria was assessed providing information about the antibiotic resistance (AR) among the groups. Based on 16S rRNA gene metagenomic analysis, over 99.78% of reads were classified as bacteria in all samples. Shannon diversity index varies from 1.22 to 2.22, with the highest bacterial population assigned to AFPE samples (1327 species). The highest microbial counts of indicator Staphylococcus spp. (STAPHY), Enterobacter spp. (ENT), and Listeria spp. (LIST), were detected in AMPE samples. Thirty percent of the selected STAPHYs, and 20.91% of Enterobacter (ENT) clones were resistant to various classes of antibiotics. The MAR index varies between 0.25 to 0.88 and was clone-, and fruit ripe stage-dependent. The results indicated that ready to eat avocados contained detectable levels of MAR bacteria, including methicillin resistant (MR)-STAPHY, which may act as a potential vector for the spread of antibiotic resistance. To achieve the increase of the production and marketing of Fuerte cultivar in Ecuador, it is vitally important to consider valuable strategies to protect the fruits at the early ripe stage in future. Thus, it is crucial to set up efficient control measures and develop coordinated strategies to guarantee the microbiological quality of the food.

Synergistic influence of probiotic and florfenicol on embryonic viability, performance, and multidrug-resistant Salmonella Enteritidis in broiler chickens

This study was conducted during the period of August 2021 to April 2022 and divided into two parts. The first part involved the isolation and characterization of Salmonella from 200 diseased broiler chickens collected from farms in Dakahlia Governorate, Egypt, with the detection of its antimicrobial susceptibility. The second experimental part involved in ovo inoculation of probiotics and florfenicol to evaluate their effects on hatchability, embryonic viability, growth performance traits and the control of multidrug-resistant Salmonella Enteritidis infections post hatching. The point prevalence of Salmonella in the internal organs of diseased chickens was 13% (26/200), including 6 serotypes: S. Enteritidis, S. Typhimurium, S. Santiago, S. Colindale, S. Takoradi and S. Daula. Multidrug resistance was found in 92% (24/26) of the isolated strains with a multiantibiotic resistance index of 0.33–0.88 and 24 antibiotic resistance patterns. The in ovo inoculation of probiotic with florfenicol showed significant improvement in the growth performance parameters compared with other groups and had the ability to prevent colonization of multidrug resistant S. Enteritidis in the majority of the experimental chicks, and the remaining chicks showed very low colonization, as detected by RT‒PCR. These findings suggested the application of in ovo inoculation techniques with both probiotics and florfenicol as a promising tool to control multidrug-resistant S. Enteritidis in poultry farms.

The potential role of migratory birds in the transmission of pathogenic Campylobacter species to broiler chickens in broiler poultry farms and live bird markets

BackgroundCampylobacter species (spp.) are one of the most important zoonotic bacteria possessing potential hazards for animal and human health worldwide. Migratory birds are implicated as significant carriers for microbes and a play very important role in the dissemination of Campylobacter to broiler chickens and their environment. The purpose of this investigation was to detect the prevalence, antibiotic resistant patterns, virulence and diversity of pathogenic Campylobacter spp. in 7 migratory bird species (Northern shoveler, Common pochard, Common teal, Northern pintail, Eared Grebe, Great Crested Grebe and Garganey) and broiler chickens that were collected from broiler poultry farms and live bird markets.ResultsThe prevalence of Campylobacter was 12.5% (25/200), of which 15% (15/100) was recovered from 5 migratory bird species only and 10% (10/100) from broiler chickens. At the level of migratory birds, eight isolates (53.3%) were Campylobacter jejuni (C. jejuni) and 7 isolates (46.7%) were Campylobacter coli (C. coli) meanwhile, in broiler chickens C. jejuni and C. coli were 50% (5/10) for each. All isolated strains had phenotypic resistance to doxycycline, while all of the isolates were susceptible to amikacin. The multidrug resistance to three, four or five antimicrobial classes was found in 72% (18/25) of the isolated strains. The multiantibiotic resistance index between the examined isolates was 0.22–0.77, with 10 antibiotic resistance patterns. The virulence of isolated Campylobacter strains (from both migratory birds and broiler chicken birds) was detected by targeting the VirB11, ciaB and iam genes which were recorded at 16%, 52% and 100%, respectively. Additionally, 100% and 84% of the antibiotic resistance genes were identified as tetA and BlaOXA-61, respectively.ConclusionsThe results of this study revealed the diversity between all the isolated strains from migratory birds and their similarity to broiler chicken isolates. The findings of the present study highlight the impact of migratory birds visiting Egypt and other countries on pathogenic Campylobacter spp. carrying pathogenic virulence and resistance genes, necessitating the application of biosecurity measures to prevent migratory birds from entering farms during their migration period.

The growing threat of antibiotic resistance in wound infections: Evidence from tertiary care in Pakistan

The present study analyzed 361 non-duplicated wound swab samples from 187 males and 174 females, ranging in age from 0 to 100 years with a mean age of 37.1?1.9 years, and to determine the prevalence of bacterial wound infections and the diversity of antibacterial susceptibility patterns of the isolated bacteria to detect the presence of unique/rare resistance types. Of these, 53.46% (193) were found to have wound infections. Most of the infected patients fell in the age group II (21-40 years). A total of 14 bacterial species were identified, with Staphylococcus aureus and Escherichia coli being the most common Gram-positive and Gram-negative bacteria, respectively. Linezolid and vancomycin were the most effective antibiotics against the isolated Gram-positive bacteria, while most Gram-negative bacteria were sensitive against colistin and polymyxin-B. Based on antibiotic resistance, 129 types of resistance were detected. Multi-resistance was detected in 157 (81.3%) bacterial strains, while 162 strains had a multi-antibiotic resistance index (MAR) of 0.2. Simpson and Shannon diversity indices indicated high bacterial diversity in the wound samples. The study provides valuable insight into the prevalence of bacterial infections in wounds and that antibiotic resistance patterns can be useful in guiding the development of effective treatment strategies.

Co-occurrence of antibiotic and metal resistance in long-term sewage sludge-amended soils: influence of application rates and pedo-climatic conditions

Urban sewage sludge (USS) is increasingly being used as an alternative organic amendment in agriculture. Because USS originates mostly from human excreta, partially metabolized pharmaceuticals have also been considered in risk assessment studies after reuse. In this regard, we investigated the cumulative effect of five annual USS applications on the spread of antibiotic-resistant bacteria (ARB) and their subsequent resistance to toxic metals in two unvegetated soils. Eventually, USS contained bacterial strains resistant to all addressed antibiotics with indices of resistance varying between 0.25 for gentamicin to 38% for ampicillin and azithromycin. Sludge-amended soils showed also the emergence of resistome for all tested antibiotics compared to non-treated controls. In this regard, the increase of sludge dose generally correlated with ARB counts, while soil texture had no influence. On the other hand, the multi-antibiotic resistance (MAR) of 52 isolates selected from USS and different soil treatments was investigated for 10 most prescribed antibiotics. Nine isolates showed significant MAR index (≥ 0.3) and co-resistance to Cd, As and Be as well. However, events including an extreme flash flood and the termination of USS applications significantly disrupted ARB communities in all soil treatments. In any case, this study highlighted the risks of ARB spread in sludge-amended soils and a greater concern with the recent exacerbation of antibiotic overuse following COVID-19 outbreak.

Effects of packaging material on microbial load and safety of ready–to-eat Voandzeia subterranean cake (Okpa)

Background: Food-borne diseases remain a big challenge to food regulatory agencies. This study evaluated the microbiological safety of Voandzeia subterranean cake also called Bambara groundnut cake (BGC) packaged with cellophane and banana leaves. Methodology: Twelve samples, 6 packaged in cellophane and 6 packaged in banana leaves were randomly selected from 6 strategic locations including, Government House Area (GHA), Eke- Awka market (EAM), UNIZIK junction (UZJ), Aroma junction (ARJ), and Nwagu junction, Agulu (NWJ). Two other samples were prepared fresh at School of Pharmacy, Agulu (SOP) which served as control. All samples were analyzed for total aerobic and fungi counts. Antimicrobial susceptibility evaluation was done on the isolates to determine their antibiogram. Results: Microbial load of V. subterranean cake packaged in cellophane and banana leaves at 24 h post-preparation were &lt;104 CFU.g-1 which is within safe limit of consumption. However, all samples packaged with banana leaves exhibited heavy contaminations with microbes at 48 h (above 105 CFU.g-1). Conversely, there were no growths of fungi in samples packaged with cellophane. The organisms: Micrococcus spp, Escherichia coli, Lactobacillus spp, S. aureus and Salmonella spp were found as contaminant. All the bacterial isolates were resistant to Co-amoxiclav and Cloxacillin. Salmonella spp isolates were resistant to all the antibiotics tested. Only the E. coli isolates had a Multi-Antibiotic Resistance Index (MARI) of less than 30%. Conclusion: The BGC packaged with cellophane are within the acceptable microbiological standard within the first day of production and compared better, microbiologically, than those packaged with banana leaves. However, the isolation of E. coli and Salmonella spp from the products questions the safety of the products and could spread Multi-Antibiotic Resistant isolates. From this study, good sanitary condition is recommended during preparation and packaging processes of V. subterranean cake with banana leaves.

Antibiotic Resistance Profiles of Bacteria Isolated from Hotspot Environments in Bahir Dar City, Northwestern Ethiopia.

BackgroundWastes generated from hotspot environments contain a wide range of antibiotics and pathogens that play a significant role in the dissemination of antibiotic-resistant bacteria in the environment. This study was carried out to isolate bacteria from hotspot environments and determine their resistance profiles to commonly used antibiotics in Bahir Dar city, Ethiopia.MethodsA cross-sectional study was conducted from October 2020 to June 2021 in Bahir Dar City. A total of 126 waste and wastewater samples were aseptically collected, transported, and processed for bacteriological isolation and susceptibility testing following standard procedures.ResultsA total of 411 bacterial isolates were recovered and the highest value of 122 (29.7%) bacterial isolates were obtained from medical wastewater samples, and the most frequently isolated bacteria were assigned to the species Escherichia coli with 82 strains (19.5%). The results revealed that the highest resistance profile of 69 (95.8%) was obtained in Staphylococcus aureus against ampicillin and 46 (86.8%) Citrobacter spp. against tetracycline. Two hundred and sixteen (52.6%) of bacteria showed multi-drug resistance and the highest multi-drug resistance was observed in Pseudomonas spp. 47 (65.3%), followed by Escherichia coli 51 (62.2%). The highest resistance profile of 12 (85.7%) and 60 (74.1%) for tetracycline were obtained from beef waste and wastewater and medical wastewater samples. The highest multi-drug resistance was recorded in isolates isolated from beef waste and wastewater samples 11 (64.7%), followed by medical wastewater samples 84 (64.1%). Even though a higher (>0.2) multi-antibiotic resistance index was found in all hotspot environments, the highest multi-antibiotic resistance index (0.477) was recorded in bacteria isolated from medical wastewater.ConclusionIt was concluded that wastes generated from hotspot environments and released in the environment contain large numbers of antibiotic-resistant, multidrug, extensively, and pan-drug-resistant bacteria. Proper waste management strategies should be established.

Occurrence and antibiogram of multidrug-resistant Salmonella enterica isolated from dairy products in Libya.

Background and Aim:Foodborne illnesses are a serious challenge to human health and the economic sector. For example, salmonellosis remains a burden in developed and developing nations. Rapid and reliable molecular methods to identify Salmonella strains are essential for minimizing human infection. This study aimed to identify Salmonella spp. in raw milk and dairy products using conventional and molecular techniques and to test the antibiotic susceptibility of the isolated strains.Materials and Methods:One hundred and thirty-one milk and dairy product samples were randomly collected from different localities in Libya. Samples were examined for the presence of Salmonella by conventional culture techniques, including cultivation in Rappaport-Vassiliadis broth and streaking on xylose lysine deoxycholate agar. Identification also used polymerase chain reaction and partial sequencing of 16S rDNA. Twenty-four antibiotics were used for the examination of antimicrobial resistance of Salmonella spp. isolates with the agar disk diffusion method (Kirby–Bauer technique). Multi-antibiotic resistance index and antibiotic resistance index (ARI)for Salmonella enterica isolates were calculated.Results:Twenty-one of 131 samples (16%) were positive for Salmonella spp. recovered from 9 (16%), 2 (11%), 4 (22.2%), and 6 (46%) samples of raw cow milk, fermented raw milk, and fresh locally made soft cheeses, Maasora and Ricotta), respectively. Samples of ice cream, milk powder, and infant formula showed no Salmonella spp. contamination. Only 9 of 21 (42.8%) isolates were confirmed as S. enterica by partial sequence 16S rDNA analysis. All isolates were resistant to amoxycillin, bacitracin, penicillin G, lincomycin, vancomycin, clindamycin, and cloxacillin with an ARI of 0.042. In contrast, all tested strains were sensitive to levofloxacin, doxycycline, and ciprofloxacin. In addition, all of the tested isolates (100%) were resistant to more than one antibiotic.Conclusion:This study demonstrated the applicability of molecular techniques, compared with conventional methods, as preferable for the identification of Salmonella in milk and dairy products and thus reduction of milk-borne transmission to the consumers. From the view of public health, isolation and identification of Salmonella multidrug-resistant strains from raw cow`s milk and locally prepared dairy products sold in the Libyan markets indicate the need to improve the handling and processing of milk and dairy products to minimize the prevalence of Salmonella, one of the most important foodborne microorganisms that cause food poisoning.

Epidemiological study on Listeria monocytogenes in Egyptian dairy cattle farms’ insights into genetic diversity of multi-antibiotic-resistant strains by ERIC-PCR

Listeria monocytogenes (L. monocytogenes) is frequently detected in ruminants, especially dairy cattle, and associated with the sporadic and epidemic outbreak of listeriosis in farms. In this epidemiological study, the prevalence, virulence, antibiotic resistance profiles, and genetic diversity of L. monocytogenes in three Egyptian dairy cattle farms were investigated. The risk factors associated with the fecal shedding of L. monocytogenes were analyzed. The L. monocytogenes strains from the three farms were categorized into distinct genotypes based on sampling site and sample type through enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). A total of 1896 samples were collected from animals, environments, and milking equipment in the three farms. Results revealed that 137 (7.23%) of these samples were L. monocytogenes positive. The prevalence of L. monocytogenes in the animal samples was high (32.1%), and the main environmental source of prevalent genotypes in the three farms was silage. For all sample types, L. monocytogenes was more prevalent in farm I than in farms II and III. Risk factor analysis showed seasonal variation in production hygiene. For all sample types, L. monocytogenes was significantly more prevalent in winter than in spring and summer. The level of L. monocytogenes fecal shedding was high likely because of increasing age, number of parities, and milk yield in dairy cattle. Two virulence genes, namely, hlyA & prfA, were also detected in 93 strains, whereas only one of these genes was found in 44 residual strains. Conversely, iap was completely absent in all strains. The strains exhibited phenotypic resistance to most of the tested antibiotics, but none of them was resistant to netilmicin or vancomycin. According to sample type, the strains from the animal samples were extremely resistant to amoxicillin (95.2%, 80/84) and cloxacillin (92.9%, 78/84). By comparison, the strains from the environmental samples were highly resistant to cefotaxime (86.95%, 20/23). Furthermore, 25 multi-antibiotic resistance (MAR) patterns were observed in L. monocytogenes strains. All strains had a MAR index of 0.22–0.78 and harbored antibiotic resistance genes, including extended-spectrum β-lactamase (blaCTX-M [92.7%] and blaDHA-1 [66.4%]), quinolones (qnrS [91.2%], qnrA [58.4%], parC [58.4%], and qnrB [51%]), macrolides (erm[B] [76.6%], erm(C) [1.5%], and msr(A) [27%]), trimethoprim (dfrD [65.7%]), and tetracyclines (tet(M) [41.6%], tet(S) [8%], and int-Tn [26.3%]). ERIC-PCR confirmed that the strains were genetically diverse and heterogeneous. A total of 137 isolated L. monocytogenes strains were classified into 22 distinct ERIC-PCR groups (A–V). Among them, ERIC E (10.2%) was the most prevalent group. These results indicated that environment and milking equipment served as reservoirs and potential transmission ways of virulent and multidrug-resistant L. monocytogenes to dairy animals, consequently posing threats to public health. Silage is the main environmental source of prevalent genotypes on all three farms. Therefore, hygienic measures at the farm level should be developed and implemented to reduce L. monocytogenes transmission inside dairy cattle farms.

Clonal diversity and spatial dissemination of multi-antibiotics resistant Staphylococcus aureus pathotypes in Southwest Nigeria

Spread of genetically diverse Staphylococcus aureus characterized with multi-antibiotic resistance and regulated by high level agr functionalities in several communities in southwest Nigeria was investigated and evaluated for infection control. Staphylococcus aureus pathotypes recovered from 256 cases including purulent pus from skin infections, soft tissue aspirates, wounds, otorrhea, eye, throat and endocervical infections were assayed for biofilm and antibiogram. Further genotyped with micro-array, mapped for geospatial distribution and evaluated for clonal diversity and functional accessory gene regulators (agr). Significant Staphylococci infection among the ages (OR:0.021, CI:0.545–1.914) and female gender with prevalence rate of MSSA (53.0%) and MRSA (1.5%) (OR:1.021, CI:0.374–1.785) were observed. More than 52.5% resistance rates to tetracycline and amoxicillin with significant median resistance were observed in all the infection cases (p = 0.001). Resistance rate of 78.8% at MIC50 32μg/ml and MIC90 128μg/ml to amoxicillin-clavulanate, and more than 40% resistance to ceftazidime, ciprofloxacin and tetracycline of MIC90 and MIC50 at 32 μg/ml were observed. Strains with multi-antibiotic resistance index above 0.83, high beta-lactamase and strong biofilm clustered into separate phylo-group. Heterogeneous t442 (wound and pus), t657 (wound), t091 (ear) and t657 (ear and wound) revealed high phylogenetic diversity. Only 4.6% pvl+ MSSA-CC1 agrI, pvl+ MSSA-CC5 (13.6%) and pvl+ MRSA-CC7 agrII (4.6%), expressed enterotoxin, leukocidins, proteases and resistance gene determinants. Livestock clonal types clustered with identified community-associated strains. Clonal dissemination of resistant pvl+ MSSA-CC1 and MRSA-CC5 encoding agr were predominant in several peri-urban communities where adequate geno-surveillance, population-target antimicrobial stewardship, extensive community structured infection control programs are needed to prevent further focal dissemination.

Antibacterial and Antioxidant Activity of Different Extracts of Some Wild Medicinal Mushrooms from Nigeria.

This study assessed the antioxidant properties and antibacterial effects of hot water, cold water, and ethanol extracts of Agaricomycetes mushrooms Trametes betulina, Pleurotus ostreatus, Agaricus bisporus var. albidus, and A. bisporus var. avellaneus against Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. The mushroom extracts were assessed for total phenolic content as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydrogen peroxide scavenging activities. The multiantibiotic resistance index (MARI) of the bacterial isolates was assessed and the antimicrobial activity of the mushroom extracts was assayed with the agar disc diffusion method. The agar well diffusion assay was performed to determine their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The test bacteria were multidrug resistant against antibiotics, with P. aeruginosa having the highest MARI followed by K. pneumoniae and E. coli. Cold water extracts showed a better antibacterial effect compared to the others and a concentration/effect correlation was observed (from 5 to 500 mg/mL). Ethanol extracts from P. ostreatus and hot water extracts from T. betulina were the most effective in inhibiting and lysing E. coli and K. pneumoniae, respectively, at a MIC and MBC of 3.90 mg/mL. The tested extracts, especially P. ostreatus extract, showed high DPPH and hydrogen peroxide scavenging potential as well as high total phenolic content compared to standard antioxidants. This study showed that the screened agaricomycetous mushrooms possess antibacterial activity against bacterial pathogens as well as antioxidant activity due to their high phenolic content.

Prevalence, virulence genes, and antimicrobial resistance of Vibrio species isolated from diseased marine fish in South China

Here, 70 potential Vibrio pathogens belonging to nine species, dominated by Vibrio harveyi, were isolated and identified from diseased aquacultured marine fish in South China. Subsequently, the prevalence of 11 virulence genes and the resistance to 15 antibiotics in these strains were determined. Most strains possessed atypical virulence genes in addition to typical virulence genes. Notably, hflk and chiA originating from V. harveyi, and flaC associated with V. anguillarum were detected in more than 40% of atypical host strains. Multidrug resistance was widespread: 64.29% strains were resistant to more than three antibiotics, and the multi-antibiotic resistance index ranged from 0.00 to 0.60. The proportions of strains resistant to the antibiotics vancomycin, amoxicillin, midecamycin, and furazolidone all exceeded 50%; nevertheless, all strains were sensitive to florfenicol, norfloxacin, and ciprofloxacin. Furthermore, both virulence genes and antibiotic resistance were more prevalent in Hainan than in Guangdong, owing to the warmer climate and longer annual farming time in Hainan. These results therefore suggest that warming temperatures and overuse of antibiotics are probably enhancing antibiotic resistance and bacterial infection. This study reveals that pathogenic Vibrio spp. with multi-antibiotic resistance are highly prevalent among marine fish in South China and thus warrant further attention. The results will provide helpful guidance for ecological regulation and local antibiotic use in the control of marine fish farming’ Vibrio diseases in South China, facilitating the implementation of national green and healthful aquaculture.

Aquaculture at the crossroads of global warming and antimicrobial resistance

In many developing countries, aquaculture is key to ensuring food security for millions of people. It is thus important to measure the full implications of environmental changes on the sustainability of aquaculture. We conduct a double meta-analysis (460 articles) to explore how global warming and antimicrobial resistance (AMR) impact aquaculture. We calculate a Multi-Antibiotic Resistance index (MAR) of aquaculture-related bacteria (11,274 isolates) for 40 countries, of which mostly low- and middle-income countries present high AMR levels. Here we show that aquaculture MAR indices correlate with MAR indices from human clinical bacteria, temperature and countries’ climate vulnerability. We also find that infected aquatic animals present higher mortalities at warmer temperatures. Countries most vulnerable to climate change will probably face the highest AMR risks, impacting human health beyond the aquaculture sector, highlighting the need for urgent action. Sustainable solutions to minimise antibiotic use and increase system resilience are therefore needed.

Prevalence of Multi-Antibiotic Resistant Escherichia coli and Klebsiella species obtained from a Tertiary Medical Institution in Oyo State, Nigeria.

Background: The development of multi-antibiotic resistant bacteria, especially Gram-negative bacteria which are the major cause of hospital-acquired infections worldwide, had been increasing. Escherichia coli and Klebsiella sp. had become more resistant to different classes of antibiotics, and the treatment of infections caused by these bacteria had developed into a challenge in both developed and developing countries. This study had determined the multi-antibiotic resistance (MAR) patterns of Escherichia coli and Klebsiella sp. isolated from clinical inpatient and outpatient samples. Method: The present study had used 50 E. coli and 48 Klebsiella sp. isolates. Antibiotic susceptibility test had been carried out by using disk diffusion method, and the interpretation of results of the zones of inhibition had accorded with Clinical Laboratory Standards Institute (CLSI). The antibiotics used had included the following: streptomycin, ciprofloxacin, erythromycin, nitrofurantoin, amikacin, gentamicin, ofloxacin, cefepime, oxacillin, colistin sulfate, cefotaxime, ceftazidime, pefloxacine, and cloxacillin. Results: E. coli and K. pneumoniae had shown high-resistance patterns. E. coli had exhibited high resistance against cloxacillin (96%), oxacillin (96%), erythromycin (88%), and most especially streptomycin (98%). Similarly, K. pneumoniae had presented a high resistance to streptomycin (88%), cloxacillin (92%), oxacillin (92%), and colistin (92%). E. coli had presented the highest multidrug resistance with a MAR index of 1.00. A total of 17 E. coli isolates had shown resistance to the 14 antibiotics tested. Conclusion: E. coli and Klebsiella sp. in clinical isolates in outpatients and inpatients in Ibadan, Western Nigeria had demonstrated high antimicrobial resistance. Thus, such condition should be considered a major public health concern, and measures must be taken to establish the sources and drivers of this problem.

Geospatial Investigation of Nigerian Honey and Detection of Anti-Enteric Biomarker.

Geospatial mapping and antibacterial biomarkers were investigated in Nigerian honey used for therapeutic purposes in several communities affected with prevalent antibiotic-resistant enteric bacilli. Randomly collected enteric bacilli from faecal samples were biotyped and phenotypically assayed for antibiotic resistance and profiled for R plasmids. R plasmid molecular weight and multiantibiotic resistance index (MARI) relatedness were evaluated for resistance among phylogroups. Honey cidal activity, time kill kinetics, and bioactive markers were determined and analysed for geospatial distribution. More than 30% enteric biotypes were resistant to cotrimoxazole, ciprofloxacin, and tetracycline at MIC ≥16 μg/ml (P=0.004). Two unrelated cluster complexes with diverse antibiotic resistance indices expressed high molecular weight plasmid (14.17 kbp) with 0.73 MARI to two classes of antibiotics. Among the resistant bacilli, only 24.3% (MIC90 500 mg/mL) and 8.1% (MBC90 1000 mg/mL) were susceptible to honey with evidence of 14.85% and 5.94% significant viable reduction at 2 × MIC to less than 2.50 Log10 CFU/mL (P < 0.05). Only alkaloids significantly regressed (P=0.028) with susceptibility of resistant bacilli significantly correlate with bacteria inhibition (r = 0.534, P=0.049) at optimal cutoff limit of 0.32 mg/ml. Antibacterial honey with significant alkaloid biomarkers was detected at 3°10′0–3°30′0E and 6°30′0–7°30′0N of Southwest Nigeria. Spatial mapping evidently indicated variation in honey physicochemical and bioactive compounds and identified geographical locations suitable for production of anti-enteric honey rich in alkaloids marker required for prevention and treatment of resistant enteric bacilli infections.

High level Multi-Resistant and Virulent Escherichia coli in Abeokuta, Nigeria

Multi-resistant Escherichia coli (E. coli) strains co-harboring virulence genes is a cause of high morbidity in Abeokuta, Nigeria. This study was designed to determine some virulent factors among enteropathogenic E. coli in Abeokuta, Nigeria. Approximately non-repetitive 102 isolates of E. coli were recovered from clinical samples from two health facilities in Abeokuta. Biotyping using API and antibiotic susceptibility was determined, and eae and flic genes were assayed by PCR. Antibiotic resistance relatedness was performed by DendroUPGMA. Results showed that 48.0% and 52.0 % were intestinal and extra-intestinal E. coli, ampicillin recorded 100% resistance, amoxycilli/clavulanic acid 64.7%, cotrimoxazole 57.8% and 56.8% resistance against cefotaxime, at MIC >16 ug/mL, 100%, 57.8%, and 50% have MIC50 to ampicillin, tetracycline, and ceftazidime, while 74.5% and 48.0% have MIC90 to ampicillin and ceftazidime. Significant rates of 4.9%, 7.8%, and 9.8% flic, eae, and flic/eae genes were found in intestinal isolates, while 2.9%, 2.0%, and 3.9% were found in extra-intestinal (P < 0.05). Two major clades of the resistant isolates reveal significant antibiotic relatedness among intestinal and extra-intestinal isolates, at 54% resistance similarities with very high multi-antibiotic resistance index of 1.0 (MARI). A high rate of undetected virulent E. coli pathotypes with high resistance could trigger unprecedented morbidity and mortality, mostly among children and the elderly.