Multiple Antibiotic Resistance Index Research Articles

Water Treatment Stage Impacts on the Occurrence of Bacteriological Indicators and their Multiple Antibiotic Resistance Index

Water Treatment Stage Impacts on the Occurrence of Bacteriological Indicators and their Multiple Antibiotic Resistance Index

Safety assessment of enterocin-producing Enterococcus strains isolated from sheep and goat colostrum

BackgroundThis study investigates the safety evaluation of enterocin-producing 11 E. mundtii and two E. faecium strains previously isolated from small livestock colostrums. Enterococcus species do not possess Generally Recognized as Safe (GRAS) status. Hence, it is critical to scrutinize enterococci’s antibiotic resistance, virulence characteristics, and biogenic amine production capabilities in order to assess their safety before using them as starter or adjunct cultures.ResultsEnterococcus strains showed susceptibility to medically significant antibiotics. Multiple-drug resistance (MDR) was found in only E. faecium HC121.4, and its multiple antibiotic resistance (MAR) index was detected to be 0.22. The tetL and aph(3')-IIIa were the most commonly found antibiotic resistance genes in the strains. However, E. mundtii strains HC56.3, HC73.1, HC147.1, and E. faecium strain HC121.4 were detected to lack any of the antibiotic resistance genes examined in this study. Only E. mundtii HC166.3 showed hemolytic activity, while none of the strains engage in gelatinase activity. The strains were identified to have virulence factor genes with a low rate. None of the virulence factor genes could be detected in E. mundtii HC26.1, HC56.3, HC73.1, HC165.3, HC166.8, and E. faecium HC121.4. The E. mundtii HC73.2 strain displayed the highest presence of virulence factor genes, namely gelE, efaAfs, cpd, and ccf. Similarly, the E. mundtii HC112.1 strain showed a significant presence of genes efaAfm, ccf, and acm. There was no decarboxylation of histidine, ornithine, or lysine seen in any of the strains. Nevertheless, E. faecium HC121.4 and HC161.1 strains could decarboxylate tyrosine, but E. mundtii HC26.1, HC56.3, HC73.1, HC73.2, HC112.1, HC147.1, HC155.2, HC165.3, HC166.3, HC166.5, and HC166.8 strains only showed a limited capacity for tyrosine decarboxylation. None of the strains possessed the hdc, odc, or ldc genes, but all of them had the tdc gene.ConclusionThe E. mundtii HC56.3 and HC73.1 strains were deemed appropriate for utilization in food production. Using the remaining 11 strains as live cultures in food production activities could pose a possible risk to consumer health.

Wastewater-based surveillance of Vibrio cholerae: Molecular insights on biofilm regulatory diguanylate cyclases, virulence factors and antibiotic resistance patterns

Vibrio cholerae is an inherent inhabitant of aquatic ecosystems. The Indian state of West Bengal, especially the Gangetic delta region is the highest cholera affected region and is considered as the hub of Asiatic cholera. V. cholerae were isolated from publicly accessible wastewater of Midnapore, West Bengal, India. Serotyping determined all isolates to be of non-O1/non-O139 serogroups. Moderate biofilm-forming abilities were noticed in most of the isolates (74.7 %) while, high biofilm formation was recorded for only 6.3 % isolates and 19 % of isolates exhibited low/non-biofilm-forming abilities. PCR-based screening of crucial diguanylate cyclases (DGCs) involved in cyclic-di-GMP-mediated biofilm signaling was performed. cdgH and cdgM were the most abundant DGCs among 93.7 % and 91.5 % of isolates, respectively. Other important DGCs, i.e., cdgK, cdgA, cdgL, and vpvC were present in 84 %, 75.5 %, 72 % and 68 % of isolates, respectively. Besides, the non-O1/non-O139 isolates were screened for the occurrence of virulence factor encoding genes. Moreover, among these non-O1/non-O139 isolates, two strains (3.17 %) harbored both ctxA and ctxB genes, which encode the cholera toxin associated with epidemic cholera. ompU was the most prevalent virulence factor, present in 24.8 % of isolates. Other virulence factors like, zot and st were found in 4.7 % and 9.5 % of isolates. Genes encoding tcp and ace were found to be PCR-negative for the isolates. Additionally, crucial virulence factor regulators, toxT, toxR and hapR were found to be PCR-positive in all the isolates. Antibiotic resistance patterns displayed further vulnerabilities with decreased sensitivity towards commonly used antibiotics with multiple antibiotic resistance index ranging between 0.37 and 0.62. The presence of cholera toxin-encoding multi-drug resistant (MDR) V. cholerae strains in environmental settings is alarming. High occurrence of DGCs are considered to encourage further investigations to use them as alternative therapeutic targets against MDR cholera pathogen due to their unique presence in bacterial systems.

Four New Sequence Types and Molecular Characteristics of Multidrug-Resistant Escherichia coli Strains from Foods in Thailand.

The presence of antibiotic-resistant Escherichia coli in food is a serious and persistent problem worldwide. In this study, 68 E. coli strains isolated from Thai food samples were characterized. Based on antibiotic susceptibility assays, 31 of these isolates (45.59%) showed multiple antibiotic resistance (MAR) index values > 0.2, indicating high exposure to antibiotics. Among these, strain CM24E showed the highest resistance (it was resistant to ten antibiotics, including colistin and imipenem). Based on genome sequencing, we identified four isolates (namely, CF25E, EF37E, NM10E1, and SF50E) with novel Achtman-scheme multi-locus sequence types (STs) (ST14859, ST14866, ST14753, and ST14869, respectively). Clermont phylogrouping was used to subtype the 68 researched isolates into five Clermont types, mainly A (51.47%) and B1 (41.18%). The blaEC gene was found only in Clermont type A, while the blaEC-13 gene was predominant in Clermont type B1. A correlation between genotypes and phenotypes was found only in Clermont type B1, which showed a strong positive correlation between the presence of an afa operon and yersiniabactin-producing gene clusters with the colistin resistance phenotype. Strain SM47E1, of Clermont type B2, carried the highest number of predicted virulence genes. In summary, this study demonstrates the pressing problems posed by the prevalence and potential transmission of antimicrobial resistance and virulence genes in the food matrix.

Whole genome sequencing of multidrug-resistant Klebsiella pneumoniae from poultry in Noakhali, Bangladesh: Assessing risk of transmission to humans in a pilot study

BackgroundMulti-drug resistant (MDR) Klebsiella pneumoniae is a public health concern due to its presence in Bangladeshi poultry products and its ability to spread resistance genes. This study genetically characterizes a distinct MDR K. pneumoniae isolate from the gut of poultry in Noakhali, Bangladesh, offering insights into its resistance mechanisms and public health impact. MethodsKlebsiella pneumoniae isolates from broiler and layer poultry were identified using biochemical and molecular analyses. Eleven isolates were tested for antibiotic sensitivity and categorized by their Multiple Antibiotic Resistance Index (MARI) profiles. The isolate with the highest MARI was selected for whole-genome sequencing using Illumina technology. The sequencing data were analyzed for genome annotation, pan-genome analysis, genome similarities, sequence type identification, and the identification of genetic determinants of resistance and virulence genes. ResultWe identified 10 MARI profiles among 11 K. pneumoniae isolates, with values ranging from 0.64 to 0.94. The highest MARI of 0.94 was found in an isolate from a layer poultry. This isolate's genome, 5401,789 base pairs long with 89.6 % coverage, showed potential inter-species dissemination, as indicated by core genome phylogenetic analysis. It possessed genes conferring resistance to fluoroquinolones, aminoglycosides, β-lactams, folate pathway antagonists, fosfomycin, macrolides, quinolones, rifamycin, tetracyclines, and polymyxins, including colistin. ConclusionPoultry serve as reservoirs for MDR K. pneumoniae, which can spread to other species and pose significant health risks. Rigorous monitoring of antibiotic use and genetic characterization of MDR bacterial isolates are essential to mitigate this threat.

Detection and characterization of multidrug resistant Escherichia coli carrying virulence gene isolated from broilers in Bangladesh.

The emergence and dissemination of multidrug resistant (MDR) bacteria pose a severe threat to public health by limiting clinical treatment and prophylactic options. This study investigates the prevalence of Escherichia coli in broilers, their phenotypic antimicrobial resistance (AMR) profiles and the presence of virulence-associated genes (VAGs) and antimicrobial resistance genes (ARGs) using polymerase chain reaction (PCR). A total of 216 pooled cloacal samples were collected from 1080 broilers across six districts of Bangladesh. Each pooled sample comprised randomly selected cloacal swabs from five birds per farm. E. coli isolates were identified using standard bacteriological approach, followed by biochemical assays and PCR. Antimicrobial susceptibility was assessed using the Kirby-Bauer disc diffusion method, and the presence of ARGs and VAGs was determined via PCR. Five selected isolates were partially sequenced for five VAGs using Sanger sequencing. A total of 177 E. coli isolates (81.94%, 95% confidence interval: 76.24%-86.53%) were identified. The isolates showed the highest resistance to ampicillin (93.79%), followed by tetracycline (91.53%), erythromycin (89.27%) and ciprofloxacin (87%). Conversely, ceftriaxone (80.79%) showed highest susceptibility, followed by gentamicin (37.29%) and neomycin (31.07%). All isolates were MDR, with a multiple antibiotic resistance indexes were <0.3. A significant percentage (16.38%) of E. coli isolates were MDR to five antimicrobial classes and harboured blaTEM, sul1, ere (A), tetA, tetB and tetC genes. The highest prevalent ARGs were blaTEM (88.14%) followed by ere (A) (83.62%) and sul 1 (72.32%). The prevalence of VAGs was astA (56.50%), iucD (31.07%), iss (21.47%), irp2 (15.82%) and cva/cvi (3.39%), respectively. This study highlights the presence of ARGs contributing to the development of MDR in E. coli carrying VAGs in broilers. Effective monitoring and surveillance of antimicrobial usage in poultry production systems are urgently required to prevent emergence and dissemination of AMR.

Multidrug-Resistant Proteus mirabilis and Other Gram-Negative Species Isolated from Native Egyptian Chicken Carcasses.

Poultry carcasses may be reservoirs for the zoonotic transmission of antimicrobial-resistant bacteria to humans and pose a major public health hazard. During the isolation of Salmonella from poultry and other foods, many of the presumptive typical Salmonella colonies on xylose lysine deoxycholate (XLD) agar were found to lack the invA gene, which is the specific target gene for Salmonella spp. Therefore, the current study aimed to estimate the prevalence and antimicrobial resistance profiles of extensively drug-resistant invA-negative non-Salmonella isolates recovered from native Egyptian chicken carcasses as presumptive Salmonella colonies on XLD agar. The non-Salmonella isolates were detected in 84% (126/150) of the examined native Egyptian chicken carcasses and classified into five genera, with prevalence rates of 64% (96/150), 14% (21/150), 6.7% (10/150), 3.3% (5/150), and 1.3% (2/150) for Proteus, Citrobacter, Shigella, Pseudomonas, and Edwardsiella, respectively. One hundred and ninety-five invA-negative, non-verified presumptive Salmonella isolates were recovered and classified at the species level into Proteus mirabilis (132/195; 67.7%), Proteus vulgaris (11/195; 5.6%), Citrobacter freundii (26/195; 13.3%), Shigella flexneri (8/195; 4.1%), Shigella sonnei (6/195; 3.1%), Shigella dysenteriae (3/195; 1.5%), Pseudomonas fluorescens (6/195; 3.1%), and Edwardsiella tarda (3/195; 1.5%). All (195/195; 100%) of these isolates showed resistance against cefaclor and fosfomycin. Additionally, these isolates showed high resistance rates of 98%, 92.8%, 89.7%, 89.2%, 89.2%, 86.7%, 80%, 78.5%, 74.4%, and 73.9% against cephalothin, azithromycin, vancomycin, nalidixic acid, tetracycline, sulfamethoxazole/trimethoprim, cefepime, gentamicin, cefotaxime, and ciprofloxacin, respectively. Interestingly, all (195/195; 100%) of the identified isolates were resistant to at least five antibiotics and exhibited an average MAR (multiple antibiotic resistance) index of 0.783. Furthermore, 73.9% of the examined isolates were classified as extensively drug-resistant, with an MAR index equal to 0.830. The high prevalence of extensively drug-resistant foodborne Proteus, Citrobacter, Shigella, Pseudomonas, and Edwardsiella isolated from native chicken carcasses poses a great hazard to public health and necessitates more monitoring and concern about the overuse and misuse of antibiotics in humans and animals. This study also recommends the strict implementation of GHP (good hygienic practices) and GMP (good manufacturing practices) in the chicken meat supply chain to protect consumer health.

Comprehensive insight on multidrug resistance and virulence genes of ESBL-producing E. coli from different surface water sources in Bangladesh

ABSTRACT Environmental dissemination of multidrug resistance (MDR) has emerged as a global concern worldwide, particularly in developing countries like Bangladesh where the waste disposal system is inadequate. The present investigation was intended to comprehend the drug resistance and virulence pattern of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) in surface water samples of Bangladesh, which are adjoining poultry farms, agricultural fields, pharmaceutical companies, and textile industries. A total of 61 E. coli with ESBL production were identified from 16 water samples based on phenotypic and genotypic approaches. MDR was observed in 96% (n = 59) of the isolates with the multiple antibiotic resistance (MAR) index in the range of 0.3–0.9. The most prevalent MDR phenotype was non-susceptible to Ampicillin–Azithromycin–Nalidixic acid, with the highest percentage in the isolates from samples near agricultural fields. The frequencies of three ESBL resistance genes were detected as blaTEM (63%), blaCTX-M-13 (60%), and blaSHV (14%). Approximately 11% of the E. coli isolates were revealed as virulent gene positive, with the predominant one (eagg) specific for Enteroaggregative E. coli (EAEC). This study implies that the aquatic environment could be a potent means of exposure and transmission of bacterial antibiotic resistance and their genetic determinants in Bangladesh.

Plastic Debris in the Aquatic Environment: An Emerging Substratum for Antimicrobial Resistant (AMR) Biofilms.

Plastic pollution has quadrupled over the past years and has become a global concern due to its direct impact on life forms. The present study analysed whether the plastic debris in aquatic environments could act as the substratum for the antimicrobial-resistant (AMR) bacteria to form biofilm for survival. We have collected various plastic debris (n = 32) from six sites of the Periyar River, the drinking water source for the entire city and one of the most polluted rivers in Kerala (India). The chemical composition of plastics was screened via FTIR analysis and found that they comprised two types, viz., polyethylene and polypropylene. Bacteria isolated from the samples were screened for the AMR characteristics towards eight different classes of antibiotics. All isolates showed 100% resistance towards colistin and obtained the MAR index value of 0.1-0.4 range. Six representative bacterial isolates with high multiple antibiotic resistance (MAR) index were selected and identified by 16sRNA sequencing as Lysinibacillus mangiferihumi, Bacillus pumilus, Bacillus safensis, Bacillus cereus, Bacillus altitudins and Bacillus pumilus. In vitro biofilm formation was experimented on the purchased plastic samples in artificial media and river water using two selected strains, Bacillus pumilus and Bacillus cereus. Significant variations were observed in biofilm growth in different media (P < 0.05) regardless of plastic types (P > 0.05). The extracellular polymeric substances (EPS) and the characteristic holes on the surface morphology were visualized in SEM analysis, thus indicating the conditioning of the plastics by the isolates for biofilm formation.

Unravelling the complex mechanisms of multidrug resistance in bovine mastitis pathogens: Insights into antimicrobial resistance genes, biofilm dynamics, and efflux systems

Mastitis remains a paramount economic threat to dairy livestock, with antibiotic resistance severely compromising treatment efficacy. This study provides an in-depth investigation into the multidrug resistance (MDR) mechanisms in bacterial isolates from bovine mastitis, emphasizing the roles of antimicrobial resistance genes (ARGs), biofilm formation, and active efflux systems. A total of 162 Staphylococci, eight Escherichia coli, and seven Klebsiella spp. isolates were obtained from 215 milk samples of clinical and subclinical mastitis cases. Antibiotic susceptibility testing identified Twenty Staphylococci (12.35 %), six E. coli (75 %) and seven Klebsiella (100 %) identified as MDR displaying significant resistance to β-lactams and tetracyclines The Multiple Antibiotic Resistance (MAR) index of these isolates ranged from 0.375 to 1.0, highlighting extensive resistance. Notably, 29 of the 33 MDR isolates produced biofilms on Congo red agar, while all exhibited biofilm formation in the Microtitre Plate assay. Critical ARGs (blaZ, blaTEM, blaCTX-M, tetM, tetA, tetB, tetC, strA/B, aadA) and efflux pump genes (acrB, acrE, acrF, emrB, norB) regulating active efflux were identified. This pioneering study elucidates the synergistic contribution of ARGs, biofilm production, and efflux pump activity to MDR in bovine mastitis pathogens. To our knowledge, this comprehensive study is the first of its kind, offering novel insights into the complex resistance mechanisms. The findings underscore the imperative need for advanced antibiotic stewardship and strategic interventions in dairy farming to curb the rise of antibiotic-resistant infections, thereby protecting both animal and public health.

A multi-pronged approach to assessing antimicrobial resistance risks in coastal waters and aquaculture systems

Antimicrobial resistance (AMR) is a global challenge that has impacted aquaculture and surrounding marine environments. In this study, a year-long monitoring program was implemented to evaluate AMR in two different aquaculture settings (i.e., open cage farming, recirculating aquaculture system (RAS)) and surrounding marine environment within a tropical coastal region. The objectives of this study are to (i) investigate the prevalence and co-occurrence of antibiotic-resistant bacteria (ARB), antibiotic resistance genes (ARGs), antibiotics (AB) and various associated chemical compounds at these study sites; (ii) explore the contributing factors to development and propagation of AMR in the coastal environment; and (iii) assess the AMR risks from different perspectives based on the three AMR determinants (i.e., ARB, ARGs and AB). Key findings revealed a distinct pattern of AMR across the different aquaculture settings, notably a higher prevalence of antibiotic-resistant Vibrio at RAS outfalls, suggesting a potential accumulation of microorganisms within the treatment system. Despite the relative uniform distribution of ARGs across marine sites, specific genes such as qepA, blaCTX−M and bacA, were found to be abundant in fish samples, especially from the RAS. Variations in chemical contaminant prevalence across sites highlighted possible anthropogenic impacts. Moreover, environmental and seasonal variations were found to significantly influence the distribution of ARGs and chemical compounds in the coastal waters. Hierarchical cluster analysis that was based on ARGs, chemical compounds and environmental data, categorized the sites into three distinct clusters which reflected strong association with location, seasonality and aquaculture activities. The observed weak correlations between ARGs and chemical compounds imply that low environmental concentrations may be insufficient for resistance selection. A comprehensive risk assessment using methodologies such as the multiple antibiotic resistance (MAR) index, comparative AMR risk index (CAMRI) and Risk quotient (RQ) underscored the complexity of AMR risks. This research significantly contributes to the understanding of AMR dynamics in natural aquatic systems and provides valuable insights for managing and mitigating AMR risks in coastal environments.

Occurrence, virulence, and AMR profile of Vibrio parahaemolyticus isolated from shellfish growing areas located along the south-west coast of India.

Vibrio parahaemolyticus is a leading cause of human gastroenteritis associated with seafood consumption. The present study aimed to investigate the occurrence and risk assessment of V. parahaemolyticus isolated from live Indian black clams, sediment, and water samples collected from shellfish harvesting areas located along the south-west coast of India. Out of the total 72 samples collected, 55.6% revealed the presence of V. parahaemolyticus; the highest occurrence was observed in shellfish samples. The presence of tdh and trh virulence genes was screened by multiplex PCR. Virulence genes could be detected in 25.8% of the strains; 19.35% of them were trh positive and 3.2% were tdh positive, while 3.2% of strains exhibited the coexistence of both virulence genes. Antimicrobial resistance (AMR) determined by the disk diffusion method revealed that 87% of the strains were multiple drug resistant and exhibited 21 diverse resistance patterns. The overall multiple antibiotic resistance (MAR) index values ranged from 0 to 0.8. To the best of our knowledge, this is the first report to document the presence of pathogenic and multidrug-resistant V. parahaemolyticus in shellfish harvesting areas of the Indian sub-continent. The study reveals possible health hazards associated with consuming shellfish harvested from the study area.

Antibiotic resistance and virulence characteristics of Vibrio vulnificus isolated from Ningbo, China.

Vibrio vulnificus (V. vulnificus) is a deadly opportunistic human pathogen with high mortality worldwide. Notably, climate warming is likely to expand its geographical range and increase the infection risk for individuals in coastal regions. However, due to the absence of comprehensive surveillance systems, the emergence and characteristics of clinical V. vulnificus isolates remain poorly understood in China. In this study, we investigate antibiotic resistance, virulence including serum resistance, and hemolytic ability, as well as molecular characteristics of 21 V. vulnificus isolates collected from patients in Ningbo, China. The results indicate that all isolates have been identified as potential virulent vcg C type, with the majority (16 of 21) classified as 16S rRNA B type. Furthermore, these isolates exhibit a high level of antibiotic resistance, with 66.7% resistance to more than three antibiotics and 61.9% possessing a multiple antibiotic resistance (MAR) index exceeding 0.2. In terms of virulence, most isolates were categorized as grade 1 in serum resistance, with one strain, S12, demonstrating intermediate sensitivity in serum resistance, belonging to grade 3. Whole genome analysis disclosed the profiles of antibiotic resistance genes (ARGs) and virulence factors (VFs) in these strains. The strains share substantial VF genes associated with adherence, iron uptake, antiphagocytosis, toxin, and motility. In particular, key VFs such as capsule (CPS), lipopolysaccharide (LPS), and multifunctional autoprocessing repeats-in-toxin (MARTX) are prevalent in all isolates. Specifically, S12 possesses a notably high number of VF genes (672), which potentially explains its higher virulence. Additionally, these strains shared six ARGs, namely, PBP3, adeF, varG, parE, and CRP, which likely determine their antibiotic resistance phenotype. Overall, our study provides valuable baseline information for clinical tracking, prevention, control, and treatment of V. vulnificus infections.

Microbiological and Molecular Investigation of Antimicrobial Resistance in Staphylococcus aureus Isolates from Western Romanian Dairy Farms: An Epidemiological Approach.

Antimicrobial therapy is the most frequently used medical intervention for bovine mastitis in the dairy industry. This study aims to monitor the extent of the antimicrobial resistance (AMR) problem in Staphylococcus aureus in the dairy industry in Western Romania. Twenty farms were selected by random sampling in a transverse epidemiological study conducted across four counties in Western Romania and divided into livestock units. This study assessed the association between the resistance genes to phenotypic expression of resistance and susceptibility. Isolates of S. aureus were identified and q-PCR reactions were used to detect antibiotic resistance genes. One hundred and fifty bovine and 20 human samples were positive for S. aureus. Twenty five percent of bovine isolates (30/120) and none(0/30) of the human isolates were methicillin-resistant S. aureus (MRSA). All isolates were susceptible to fosfomycin, ciprofloxacin, netilmicin, and resistant to ampicillin and penicillin. S. aureus isolates regarded as phenotypically resistant (R) were influenced by the origin of the samples (human versus bovine, χ2 = 36.510, p = 0.013), whether they were methicillin-resistant S. aureus (χ2 = 108.891, p < 0.000), the county (χ2 = 103.282, p < 0.000) and farm of isolation (χ2 = 740.841, p < 0.000), but not by the size of the farm (χ2 = 65.036, p = 0.306). The multiple antibiotic resistance index was calculated for each sample as the number regarded as phenotypically resistant (R)/total antibiotics tested (MARI = 0.590 ± 0.023) was significantly higher (p < 0.000) inmethicillin-resistant S. aureus (0.898 ± 0.019) than non-methicillin-resistant S. aureus (0.524 ± 0.024) isolates. For the antibiotics tested, the total penetrance (P%) of the resistance genes was 59%, 83% for blaZ, 56% for cfr, 50% for erm(B), 53% for erm(C), 57% for mecA and 32% for tet(K). Penetrance can be used as a parameter for guidance towards a more accurate targeting of chemotherapy. P% in S. aureus was strongly positively correlated with the multiple antibiotic resistance index (r = +0.878, p < 0.000) with the potential to use the same limit value as an antibiotic management decision criterion. Considering cow mastitis, the penetrance value combined with the multiple antibiotic resistance index suggests that penetrance could serve as a useful parameter for more precise targeting of chemotherapy for S. aureus.

Silver nanoparticle with potential antimicrobial and antibiofilm efficiency against multiple drug resistant, extensive drug resistant Pseudomonas aeruginosa clinical isolates

BackgroundThe study aims to investigate the effect of combining silver nanoparticles (AGNPs) with different antibiotics on multi-drug resistant (MDR) and extensively drug resistant (XDR) isolates of Pseudomonas aeruginosa (P. aeruginosa) and to investigate the mechanism of action of AGNPs.MethodsAGNPs were prepared by reduction of silver nitrate using trisodium citrate and were characterized by transmission electron microscope (TEM) in addition to an assessment of cytotoxicity. Clinical isolates of P. aeruginosa were collected, and antimicrobial susceptibility was conducted. Multiple Antibiotic Resistance (MAR) index was calculated, and bacteria were categorized as MDR or XDR. Minimum inhibitory concentration (MIC) of gentamicin, ciprofloxacin, ceftazidime, and AGNPs were determined. The mechanism of action of AGNPs was researched by evaluating their effect on biofilm formation, swarming motility, protease, gelatinase, and pyocyanin production. Real-time PCR was performed to investigate the effect on the expression of genes encoding various virulence factors.ResultsTEM revealed the spherical shape of AGNPs with an average particle size of 10.84 ± 4.64 nm. AGNPS were safe, as indicated by IC50 (42.5 µg /ml). The greatest incidence of resistance was shown against ciprofloxacin which accounted for 43% of the bacterial isolates. Heterogonous resistance patterns were shown in 63 isolates out of the tested 107. The MAR indices ranged from 0.077 to 0.84. Out of 63 P. aeruginosa isolates, 12 and 13 were MDR and XDR, respectively. The MIC values of AGNPs ranged from 2.65 to 21.25 µg /ml. Combination of AGNPs with antibiotics reduced their MIC by 5–9, 2–9, and 3-10Fold in the case of gentamicin, ceftazidime, and ciprofloxacin, respectively, with synergism being evident. AGNPs produced significant inhibition of biofilm formation and decreased swarming motility, protease, gelatinase and pyocyanin production. PCR confirmed the finding, as shown by decreased expression of genes encoding various virulence factors.ConclusionAGNPs augment gentamicin, ceftazidime, and ciprofloxacin against MDR and XDR Pseudomonas isolates. The efficacy of AGNPs can be attributed to their effect on the virulence factors of P. aeruginosa. The combination of AGNPs with antibiotics is a promising strategy to attack resistant isolates of P. aeruginosa.

Multiple antibiotic resistance index of Klebsiella pneumoniae isolated from clinical specimens in a tertiary hospital in Benin City, Nigeria

Multiple antibiotic resistance index of Klebsiella pneumoniae isolated from clinical specimens in a tertiary hospital in Benin City, Nigeria

Analysis of molecular epidemiological characteristics and antimicrobial susceptibility of vancomycin-resistant and linezolid-resistant Enterococcus in China

BackgroundThis study investigates the distribution and characteristics of linezolid and vancomycin susceptibilities among Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) and explores the underlying resistance mechanisms.MethodsA total of 2842 Enterococcus clinical isolates from patients were retrospectively collected, and their clinical data were further analyzed. The minimum inhibitory concentrations (MICs) of vancomycin and linezolid were validated by broth dilution method. The resistance genes optrA, cfr, vanA, vanB and vanM were investigated using polymerase chain reaction (PCR). Housekeeping genes and resistance genes were obtianed through whole-genome sequencing (WGS).ResultsOf the 2842 Enterococcus isolates, 88.5% (2516) originated from urine, with E. faecium accounted for 60.1% of these. The vanA gene was identified in 27/28 vancomycin resistant Enterococcus (VRE) isolates, 4 of which carried both vanA and vanM genes. The remaining strain was vanM positive. The optrA gene was identified in all E. faecalis isolates among linezolid resistant Enterococcus (LRE). E. faecium showed a higher multiple antibiotic resistance index (MAR index) compared to E. faecalis. The multi-locus sequence typing (MLST) showed the sequence type of E. faecium mainly belongs to clonal complex (CC) 17, nearly E. faecalis isolates analyzed were differentiated into 7 characteristics of sequence types (STs), among which ST16 of CC16 were the major lineage.ConclusionUrine was the primary source of VRE and LRE isolates in this study. E. faecium showed higher levels of resistance compared to E. faecalis. OptrA gene was detected in 91.6% of LRE, which could explain linezolid resistance, and van genes were detected in all vancomycin resistant Enterococcus strains, while vanA was a key resistance mechanism in VRE identified in this study.

Enterobacteriaceae Antibiotic Resistance Identification in Slender-billed Gull Species Migrating to Libya

The Libyan coast has various types of wetlands that are passed by many migratory birds throughout the year, however, studies on bacterial isolation from these birds in Lybia are scarce. The present study aimed to isolate enteropathogenic bacterial species from the seagulls that migrated to the Libyan coast as well as identification of the antibiotics that are resistant to these bacteria. A total of 50 fresh fecal samples were collected from slender-billed gulls in January 2023 at Farwa Island near the city of Zuwara, in Western Libya. Bacteria were isolated by conventional culturing method, identified using the Enterosystem 18R, and antibiotic susceptibility tests were conducted on the isolated bacteria. The results revealed the isolation of 46 bacteria, but only 32 of them were identified using biochemical tests. These identified bacteria belong to six species of Enterobacteriacae, namely Citrobacter (C.) freundii, Pantoea (P.) agglomerans, Escherichia (E.) coli, Enterobacter (En.) cloacae, Serratia liquifaciens, and Proteus mirabilis, with percentages of 53.125%, 31.25%, 6.25%, 3.125%, 3.125%, and 3.125%, respectively. All isolated bacteria were 100% sensitive to gentamicin (10 µg) and ciprofloxacin (5 µg). The highest resistance result was observed against the antibiotic cefoxitin (30 µg), with both C. freundii (5 samples) and P. agglomerans (4 samples). Resistance was observed in 5 samples of C. freundii and 4 samples of P. agglomerans out of 11 samples. Resistance to antibiotics, such as azithromycin (15 µg), ceftriaxone (30 µg), and ampicillin (10 µg), was also noted in a few isolates. The results indicated that C. freundii was the most antibiotic-resistant bacterial species isolated in this study. The highest multiple antibiotic resistance index was demonstrated by bacteria C. freundii, P. agglomerans, and En. cloacae, with a value of 0.33 for each of them. In conclusion, slender-billed gulls carry multi-drug-resistant bacteria. The study recommends the implementation of a national program to survey antibiotic-resistant bacteria, determine their prevalence, and assess the presence of antibiotic residues in animal-derived food. Furthermore, the present study advises expanding scientific studies on risk analysis, and antibiotic alternatives in migratory birds.

Multidrug Resistance And Susceptibility Profile of Outpatient Escherichia Coli Clinical Isolates

Background: To gain better understanding about the impact of antibiotic selection pressure on antimicrobial resistance (AMR) in Nigeria, it is important to conduct and review from time to time the susceptibility profile of clinically important bacteria including Escherichia coli. This study investigated the susceptibility and multidrug resistance profile of E. coli isolates from urine samples of patients who received outpatient’s medical services in Abakaliki. Methods: A total of 50 non-duplicate clinical isolates of E. coli from the microbiology laboratory section of a tertiary hospital in Abakaliki, Nigeria was recruited for this study. Susceptibility studies were determined using amoxicillin (20 µg), ceftazidime (30 µg), ciprofloxacin (5 µg), gentamicin (10 µg), nitrofurantoin (300 µg) and erythromycin acid (15 µg) by the modified Kirby–Bauer disk diffusion method. All susceptibility studies were carried out as per the guidelines of the Clinical Laboratory Standard Institute (CLSI). Multidrug resistance was evaluated by the multiple antibiotic resistance index (MARI) calculation. Results: The E. coli isolates showed reduced susceptibility to gentamicin (75%), ciprofloxacin (85%), nitrofurantoin (95%), amoxicillin (100%), erythromycin (100%) and ceftazidime (95%). A total of 12 E. coli isolates were multidrug resistant to clinically important antibiotics in the classes: cephalosporins, macrolides, aminoglycosides, fluoroquinolones, penicillins and nitrofurantoin. Conclusion: We report a high rate of antibiotic resistance of outpatients E. coli isolates which concurrently showed multidrug resistance to important antibiotic classes. These findings have clinical significance and provide a benchmark for future studies on the susceptibility pattern of clinical isolates in Nigeria. This preliminary study reiterates the need to reinvigorate antibiotic stewardship in our local hospitals so as to preserve the clinical efficacy of available antibiotics since it takes years for a new antibiotic to be developed.

Unveiling the ecological landscape of bacterial β-lactam resistance in Delhi-national capital region, India: An emerging health concern

Inappropriate antibiotic use not only amplifies the threat of antimicrobial resistance (AMR), moreover exacerbates the spread of resistant bacterial strains and genes in the environment, underscoring the critical need for effective research and interventions. Our aim is to assess the prevalence and resistance characteristics of β-lactam resistant bacteria (BLRB) and β-lactamase resistant bacterial genes (BLRBGs) under various environmental conditions within Delhi NCR, India. Using a culture-dependent method, we isolated 130 BLRB from 75 different environmental samples, including lakes, ponds, the Yamuna River, agricultural soil, aquatic weeds, drains, dumping yards, STPs, and gaushalas. Tests for antibiotic susceptibility were conducted in addition to phenotypic and genotypic identification of BLs and integron genes. The water and sediment samples recorded an average bacterial abundance of 3.6 × 106 CFU/mL and an average ampicillin-resistant bacterial count of 2.2 × 106 CFU/mL, which can be considered a potent reservoir of BLRB and BLRBGs. The majority of the BLRB discovered are opportunistic pathogens from the Bacillus, Aeromonas, Pseudomonas, Enterobacter, Escherichia, and Klebsiella genera, with Multiple Antibiotic Resistance (MAR) index ≥0.2 against a wide variety of β-lactams and β-lactamase (BLs) inhibitor combinations. The antibiotic resistance pattern was similar in the case of bacteria isolated from STPs. Meanwhile, bacteria isolated from other sources were diverse in their antibiotic resistance profile. Interestingly, we discovered that 10 isolates of various origins produce both Extended Spectrum BLs and Metallo BLs, as well as found harboring blaTEM, blaCTX, blaOXA, blaSHV, int-1, and int-3 genes. Enterobacter cloacae (S50/A), a common nosocomial pathogen isolated from Yamuna River sediment samples at Nizamuddin point, possesses three BLRBGs (blaTEM, blaCTX, and blaOXA) and a MAR index of 1.0, which is a major cause for concern. Therefore, identifying the source, origin and dissemination of BLRB and BLRGs in the environment is of the utmost importance for designing effective mitigation approaches to reduce a load of antimicrobial resistance factors in the environmental settings.