Using MALDI-TOF Mass Spectrometry to Identify Drug Resistant Staphylococcal Isolates from Nonhospital Environments in Brunei Darussalam. - Related Documents




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549301.0000Using MALDI-TOF Mass Spectrometry to Identify Drug Resistant Staphylococcal Isolates from Nonhospital Environments in Brunei Darussalam. Drug resistant bacteria have been a growing threat to the community and hospitals due to the misuse of antibiotics by humans, industrialization, and lack of novel antimicrobials currently available. Little is known about the prevalence of drug resistant bacteria in nonhealthcare environments in Brunei Darussalam and about how antibiotic resistant genes are transferred within these environments. Human contact points from different types of environments in Brunei Darussalam, varying from urban to jungle settings, were swabbed and cultured onto selective media to isolate staphylococci bacteria before performing antimicrobial susceptibility testing on the isolates. The identity of the isolates was determined using MALDI-TOF mass spectrometry (MS). Staphylococci isolates resistant to oxacillin were further tested for their minimum inhibitory concentration (MIC). PCR analysis of the mecA gene, a gene that confers resistance to oxacillin, is done to determine the level of resistance to oxacillin. Ten different staphylococcal species were identified by MALDI-TOF-MS analysis. Out of the 36 staphylococci isolates, 24 were resistant to multiple antibiotics including two isolates which were oxacillin resistant. Some staphylococci isolates had similar antibiotic resistance profiles to other staphylococci isolates of different species in the same location. This work provides the first-ever evidence of drug resistant staphylococci in the nonhospital environment in Brunei Darussalam.201627127505
550310.9992Molecular characterization of multi-drug resistant coagulase negative cocci in non-hospital environment. Antibiotic resistance crisis occasioned by sporadic appearance of multi-drug resistance (MDR) in human pathogens to clinically applied antimicrobials is a serious threat to global health. In this study, we investigated the drug resistant phenotype of Gram-positive cocci isolates from environment. Staphylococcus capitis and Staphylococcus haemolyticus colonies were isolated on mannitol-salt agar plates supplemented with tetracycline. Antibiotic susceptibility profile of the isolates via minimum inhibitory concentration (MIC) determination was examined. Isolates showed decreased sensitivity to clinically applied antimicrobial agents: tetracycline, kanamycin, erythromycin, norfloxacin, teicoplanin, and ampicillin. Genomic analysis demonstrated the presence of multiple antibiotic resistant genes in these bacteria, suggesting the origin of the multiple antimicrobials resistant phenotype. Tetracycline resistance of these isolates was transduced to Staphylococcus aureus-RN4220 strain. These findings indicate the presence of multiple antimicrobials resistant S. capitis and S. haemolyticus strain in a non-hospital setting. Moreover, the presence of plethora of genes responsible for MDR suggest that these strains could present potential threat to human health by serving as reservoir for lateral transference of antimicrobial resistance conferring foreign genetic elements to other clinically relevant pathogens.201931231110
577320.9992LBJMR medium: a new polyvalent culture medium for isolating and selecting vancomycin and colistin-resistant bacteria. BACKGROUND: Multi-drug resistant bacteria are a phenomenon which is on the increase around the world, particularly with the emergence of colistin-resistant Enterobacteriaceae and vancomycin-resistant enterococci strains. The recent discovery of a plasmid-mediated colistin resistance with the description of the transferable mcr-1 gene raised concerns about the need for an efficient detection method for these pathogens, to isolate infected patients as early as possible. The LBJMR medium was developed to screen for all polymyxin-resistant Gram-negative bacteria, including mcr-1 positive isolates, and vancomycin-resistant Gram-positive bacteria. RESULTS: The LBJMR medium was developed by adding colistin sulfate salt at a low concentration (4 μg/mL) and vancomycin (50 μg/mL), with glucose (7.5 g/L) as a fermentative substrate, to a Purple Agar Base (31 g/L). A total of 143 bacterial strains were used to evaluate this universal culture medium, and the sensitivity and specificity of detection were 100% for the growth of resistant strains. 68 stool samples were cultured on LBJMR, and both colistin-resistant Gram-negative and vancomycin-resistant Gram-positive strains were specifically detected. CONCLUSIONS: The LBJMR medium is a multipurpose selective medium which makes it possible to identify bacteria of interest from clinical samples and to isolate contaminated patients in hospital settings. This is a simple medium that could be easily used for screening in clinical microbiology laboratories.201729169321
556430.9992Epidemiology of the colonization and acquisition of methicillin-resistant staphylococci and vancomycin-resistant enterococci in dogs hospitalized in a clinic veterinary hospital in Spain. Antibiotic resistance is one of the biggest threats to human and animal health. Methicillin-resistant Staphylococcus spp. (MRS) and vancomycin-resistant Enterococcus spp. (VRE) are of increasing importance in hospital and/or nosocomial infections and represent a potential risk of transmission to humans from infected or colonized companion animals. Studies on the risk factors associated with colonization by multiresistant bacteria in animals are scarce. The present study aimed to estimate the prevalence and incidence of MRS and VRE in canine patients hospitalized in a veterinary hospital and to identify the risk factors for its acquisition and persistence. Nasal and perianal swabs were obtained from 72 dogs. Antimicrobial susceptibility assays and molecular detection of mecA and van genes were performed. A prevalence of 13.9% and incidence of 26.5% was observed in dogs colonized by MRS at hospital admission and release, respectively, higher values than those described in most veterinary studies. Thirty-five Staphylococcus isolates had mecA gene and showed higher resistance levels to most of the antimicrobials evaluated. Previous and concomitant use of antibiotics and corticosteroids has been associated with an increase in MRS colonization. The use of antibiotics in other animals living with the canine patients has also been identified as an associated factor, suggesting cross transmission. The presence of van-resistant genes from Enterococcus spp. was not detected. Pets should be considered possible vehicles of transmission and reservoirs for MRS bacteria and veterinary hospitals should be considered high-risk environments for the occurrence and spread of nosocomial infections and resistant bacteria.202032535110
553440.9992Antibiotic resistance in faecal microbiota of Greek healthy infants. Increasing use of antibiotics for the treatment of infectious diseases and also for non-therapeutic reasons (agriculture, animal husbandry and aquaculture) has led to the increasing incidence of antibiotic resistance and the ineffectiveness of antimicrobial treatment. Commensal intestinal bacteria are very often exposed to the selective pressure of antimicrobial agents and may constitute a reservoir of antibiotic resistance determinants that can be transferred to pathogens. The present study aimed to investigate the antibiotic susceptibility profile and the presence of selected resistance genes in cocci isolated from the faecal microbiota of 35 healthy, full-term infants at 4, 30 and 90 days after delivery. A total of 148 gram-positive, catalase-negative cocci were isolated and tested for susceptibility to 12 different antibiotics by disk-diffusion technique. Multiplex PCR analysis was performed for the identification of Enterococcus spp. isolates and the simultaneous detection of vancomycin-resistance genes. PCR-based methodology was used also for identification of tetracycline and erythromycin resistance determinants. Identification results indicated E. faecalis as the predominant species (81 strains), followed by E. faecium, E. casseliflavus/E. flavescens and E. gallinarum. High prevalence of resistance to tetracycline (39.9%), erythromycin (35.1%), vancomycin (19.6%) and to nucleic acid synthesis inhibitors was detected. PCR data revealed 24 out of 52 erythromycin-resistant isolates carrying the ermB gene and 32 out of 59 tetracycline-resistant strains carrying tet genes, with tet(L) determinant being the most frequently detected. Only intrinsic vancomycin resistance (vanC1 and vanC2/C3) was reported among tested isolates. In conclusion, erythromycin and tetracycline acquired resistant traits are widespread among faecal cocci isolates from Greek, healthy infants under no apparent antimicrobial selective pressure.201021831766
196650.9992Identification and characterization of vancomycin-resistant Staphylococcus aureus in hospital wastewaters: evidence of horizontal spread of antimicrobial resistance. Antibiotic resistance has become a major threat to human health around the world, but its spread through the aquatic environment has been often overlooked. This study aimed to determine the occurrence of vancomycin-resistant Staphylococcus aureus in hospital wastewaters and their transmission into public water bodies in Kerala, India. A total of 113 S. aureus were isolated from three hospital effluents in Kerala, India. Standard disc diffusion and the strip method were used for antibiotic susceptibility testing and minimum inhibitory concentration detection. Plasmid-mediated vancomycin resistance was confirmed by plasmid curing and conjugation; resistant genes were detected by the polymerase chain reaction (PCR). Nearly 76% of S. aureus isolates were resistant to β-lactams, chloramphenicol, macrolides, aminoglycosides, and glycopeptide class of antibiotics. Among the vancomycin-resistant Staphylococcus aureus (VRSA) isolates, the prevalence rates of vanA and vanB resistance-encoding genes were 46.5 and 59.3%, respectively. Through the broth mating method, vanA gene was successfully transferred from VRSA donor to vancomycin-sensitive S. aureus. The study strongly indicates the contamination of water bodies with antibiotic-resistant bacteria from hospital discharges, their dissemination and possible transfer to microbes in the aquatic environment, posing a serious threat for public health.202134665771
550560.9992Concordance between Antimicrobial Resistance Phenotype and Genotype of Staphylococcus pseudintermedius from Healthy Dogs. Staphylococcus pseudintermedius, a common commensal canine bacterium, is the main cause of skin infections in dogs and is a potential zoonotic pathogen. The emergence of methicillin-resistant S. pseudintermedius (MRSP) has compromised the treatment of infections caused by these bacteria. In this study, we compared the phenotypic results obtained by minimum inhibitory concentration (MICs) for 67 S. pseudintermedius isolates from the skin of nine healthy dogs versus the genotypic data obtained with Nanopore sequencing. A total of 17 antibiotic resistance genes (ARGs) were detected among the isolates. A good correlation between phenotype and genotype was observed for some antimicrobial classes, such as ciprofloxacin (fluoroquinolone), macrolides, or tetracycline. However, for oxacillin (beta-lactam) or aminoglycosides the correlation was low. Two antibiotic resistance genes were located on plasmids integrated in the chromosome, and a third one was in a circular plasmid. To our knowledge, this is the first study assessing the correlation between phenotype and genotype regarding antimicrobial resistance of S. pseudintermedius from healthy dogs using Nanopore sequencing technology.202236421269
547970.9992Novel linezolid resistance plasmids in Enterococcus from food animals in the USA. OBJECTIVES: To sequence the genomes and determine the genetic mechanisms for linezolid resistance identified in three strains of Enterococcus isolated from cattle and swine caecal contents as part of the US National Antimicrobial Resistance Monitoring System (NARMS) surveillance programme. METHODS: Broth microdilution was used for in vitro antimicrobial susceptibility testing to assess linezolid resistance. Resistance mechanisms and plasmid types were identified from data generated by WGS on Illumina® and PacBio® platforms. Conjugation experiments were performed to determine whether identified mechanisms were transmissible. RESULTS: Linezolid resistance plasmids containing optrA were identified in two Enterococcus faecalis isolates and one Enterococcus faecium. The E. faecium isolate also carried the linezolid resistance gene cfr on the same plasmid as optrA. The linezolid resistance plasmids had various combinations of additional resistance genes conferring resistance to phenicols (fexA), aminoglycosides [spc and aph(3')-III] and macrolides [erm(A) and erm(B)]. One of the plasmids was confirmed to be transmissible by conjugation, resulting in linezolid resistance in the transconjugant. CONCLUSIONS: To the best of our knowledge, this is the first identification of linezolid resistance in the USA in bacteria isolated from food animals. The oxazolidinone class of antibiotics is not used in food animals in the USA, but the genes responsible for resistance were identified on plasmids with other resistance markers, indicating that there may be co-selection for these plasmids due to the use of different antimicrobials. The transmissibility of one of the plasmids demonstrated the potential for linezolid resistance to spread horizontally. Additional surveillance is necessary to determine whether similar plasmids are present in human strains of Enterococcus.201830272180
593880.9992Characterization of Mechanisms Lowering Susceptibility to Flumequine among Bacteria Isolated from Chilean Salmonid Farms. Despite their great importance for human therapy, quinolones are still used in Chilean salmon farming, with flumequine and oxolinic acid currently approved for use in this industry. The aim of this study was to improve our knowledge of the mechanisms conferring low susceptibility or resistance to quinolones among bacteria recovered from Chilean salmon farms. Sixty-five isolates exhibiting resistance, reduced susceptibility, or susceptibility to flumequine recovered from salmon farms were identified by their 16S rRNA genes, detecting a high predominance of species belonging to the Pseudomonas genus (52%). The minimum inhibitory concentrations (MIC) of flumequine in the absence and presence of the efflux pump inhibitor (EPI) Phe-Arg-β-naphthylamide and resistance patterns of isolates were determined by a microdilution broth and disk diffusion assays, respectively, observing MIC values ranging from 0.25 to >64 µg/mL and a high level of multi-resistance (96%), mostly showing resistance to florfenicol and oxytetracycline. Furthermore, mechanisms conferring low susceptibility to quinolones mediated by efflux pump activity, quinolone target mutations, or horizontally acquired resistance genes (qepA, oqxA, aac(6')-lb-cr, qnr) were investigated. Among isolates exhibiting resistance to flumequine (≥16 µg/mL), the occurrence of chromosomal mutations in target protein GyrA appears to be unusual (three out of 15), contrasting with the high incidence of mutations in GyrB (14 out of 17). Bacterial isolates showing resistance or reduced susceptibility to quinolones mediated by efflux pumps appear to be highly prevalent (49 isolates, 75%), thus suggesting a major role of intrinsic resistance mediated by active efflux.201931847389
555290.9992Plasmid-Mediated Fluoroquinolone Resistance Genes in Quinolone-Susceptible Aeromonas spp. Phenotypes Isolated From Recreational Surface Freshwater Reservoir. Aeromonas spp. are recognized as opportunistic pathogens causing diseases. Infections in humans can result mainly in gastrointestinal and wound diseases with or without progression to septicemia. Although Aeromonas spp. are not known uropathogens and they rarely cause urinary tract infection, we hypothesize that the presence of these bacteria in the water and the contact during, e.g., recreational and bathing activity can create the conditions for the colonization of the human body and may result to diseases in various locations, including the urinary tract. Our study presents the occurrence of aeromonad fluoroquinolone-susceptible phenotypes with the presence of plasmid-mediated fluoroquinolone resistance (PMQR) genes in a natural freshwater reservoir occasionally used for recreational activities. Sixty-nine isolates collected during the bathing period were identified by mass spectrometry and screened for the presence of fluoroquinolone-resistant phenotypes and genotypes. Fluoroquinolone susceptibility was determined as minimal inhibitory concentration values. PMQR qnr genes were detected by PCR. Isolates comprising eight species, namely, mainly Aeromonas veronii (50.7% isolates) and Aeromonas media (24.6% isolates) and rarely Aeromonas eucrenophila, Aeromonas caviae, Aeromonas bestiarum, Aeromonas ichthiosmia, and Aeromonas hydrophila, were selected. All isolates were phenotypically susceptible either to ciprofloxacin or levofloxacin. Unexpectedly, at least one to three of the PMQR genes were detected in 42.0% of the fluoroquinolone-susceptible Aeromonas spp. phenotypes. Mainly the qnrS (34.8% isolates) and qnrA (14.5% isolates) determinants were detected. In conclusion, the freshwater reservoir occasionally used for bathing was tainted with aeromonads, with a high occurrence of opportunistic pathogens such as A. veronii and A. media. MALDI-TOF MS is a powerful technique for aeromonad identification. Our data reveals the mismatch phenomenon between fluoroquinolone-susceptible aeromonad phenotypes and the presence of plasmid-mediated qnr resistance genes. It suggests that phenotypically susceptible bacteria might be a potential source for the storage and transmission of these genes. The exposure during, e.g., a recreational activity may create the potential risk for causing infections, both diagnostically and therapeutically difficult, after expressing the resistance genes and quinolone-resistant strain selection.202235646727
1695100.9992Presence of the blaTEM Gene in Commensal Neisseria spp.: A Possible Cause for the Acquired Drug Resistance Among Pathogenic Respiratory Bacteria. Background The oral microbiome consists of various bacterial genera, with Neisseria spp. being a prominent part of this niche. While Neisseria gonorrhoeae and Neisseria meningitidis are human-restricted pathogens, non-pathogenic Neisseria species like Neisseria sicca, Neisseria perflava, etc., are primarily commensals that can also behave as opportunistic pathogens. With increasing penicillin resistance in commensal Neisseria, there is a concern that these bacteria might harbor resistance genes that can be transferred to other pathogens. This study aimed to characterize the blaTEM gene (encodes for the plasmid-mediated β-lactamase enzyme that hydrolyzes the β-lactam ring) of commensal Neisseria spp. isolated from respiratory samples. Methodology The research was conducted in the Department of Clinical Microbiology at Sri Ramachandra University, Chennai. The specimens used were sputum and throat swabs, which were subjected to a series of phenotypic methods and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) for speciation. The antibiogram was determined using the Kirby-Bauer disk diffusion method, and a PCR assay was utilized to identify the blaTEM( )gene responsible for β-lactamase production. Results Out of 274 processed samples, 65 unique commensal Neisseria spp. were identified. The study highlighted the presence of the blaTEM gene in 93.9% (61) of the isolates, which is responsible for β-lactamase production. All isolates exhibited resistance to penicillin. Most blaTEM-positive commensal Neisseria spp. were susceptible to cefuroxime (83.6%), ceftriaxone (85.2%), and cefotaxime (85.2%). The high prevalence of the blaTEM gene in commensal Neisseria is alarming. The gene, found on plasmids, could potentially transfer to other related species like Neisseria gonorrhoeae and Neisseria meningitidis, as well as other Gram-negative bacilli. Conclusion The presence of resistance genes in commensal bacteria is of concern, as they might be reservoirs for resistance transfer to pathogenic strains. The study emphasizes the importance of continuous monitoring and deeper investigations into commensal bacteria, emphasizing the need for a broader community screening approach to understand resistance mechanisms in the normal microbiome.202338146567
5600110.9992The Characterization and Beta-Lactam Resistance of Staphylococcal Community Recovered from Raw Bovine Milk. Staphylococci is an opportunistic bacterial population that is permanent in the normal flora of milk and poses a serious threat to animal and human health with some virulence factors and antibiotic-resistance genes. This study was aimed at identifying staphylococcal species isolated from raw milk and to determine hemolysis, biofilm, coagulase activities, and beta-lactam resistance. The raw milk samples were collected from the Düzce (Türkiye) region, and the study data represent a first for this region. The characterization of the bacteria was performed with MALDI-TOF MS and 16S rRNA sequence analysis. The presence of coa, icaB, blaZ, and mecA was investigated with PCR. A nitrocefin chromogenic assay was used for beta-lactamase screening. In this context, 84 staphylococci were isolated from 10 different species, and the dominant species was determined as S. aureus (32.14%). Although 32.14% of all staphylococci were positive for beta hemolysis, the icaB gene was found in 57.14%, coa in 46.42%, mecA in 15.47%, and blaZ in 8.33%. As a result, Staphylococcus spp. strains that were isolated from raw milk in this study contained some virulence factors at a high level, but also contained a relatively low level of beta-lactam resistance genes. However, considering the animal-environment-human interaction, it is considered that the current situation must be monitored constantly in terms of resistance concerns. It must not be forgotten that the development of resistance is in constant change among bacteria.202336978423
5597120.9992Prevalence of macrolide-lincosamide-streptogramin resistant lactic acid bacteria isolated from food samples. Lactic acid bacteria (LAB) being a reservoir of antibiotic resistance genes, tend to disseminate antibiotic resistance that possibly pose a threat to human and animal health. Therefore, the study focuses on the prevalence of macrolide-lincosamide-streptogramin- (MLS) resistance among LAB isolated from various food samples. Diverse phenotypic and genotypic MLS resistance were determined among the LAB species (n = 146) isolated from fermented food products (n = 6) and intestine of food-producing animals (n = 4). Double disc, triple disc diffusion and standard minimum inhibitory concentration (MIC) tests were evaluated for phenotypic MLS resistance. Specific primers for MLS resistance genes were used for the evaluation of genotypic MLS resistance and gene expressions using total RNA of each isolate at different antibiotic concentrations. The isolates identified are Levilactobacillus brevis (n = 1), Enterococcus hirae (n = 1), Limosilactobacillus fermentum (n = 2), Pediococcus acidilactici (n = 3), Enterococcus faecalis (n = 1). The MIC tests along with induction studies displayed cMLS(b), L phenotype, M phenotype, KH phenotype, I phenotype resistance among MLS antibiotics. Genotypic evaluation tests revealed the presence of ermB, mefA/E, msrA/B and msrC genes. Also, gene expression studies displayed increased level of gene expression to the twofold increased antibiotic concentrations. In the view of global health concern, this study identified that food samples and food-producing animals represent source of antibiotic resistant LAB that can disseminate resistance through food chain. This suggests the implementation of awareness in the use of antibiotics as growth promoters and judicious use of antibiotics in veterinary sectors in order to prevent the spread of antibiotic resistance.202336712199
1599130.9992Colistin Resistance Genes in Broiler Chickens in Tunisia. Colistin is a polymyxin antibiotic that has been used in veterinary medicine for decades, as a treatment for enterobacterial digestive infections as well as a prophylactic treatment and growth promoter in livestock animals, leading to the emergence and spread of colistin-resistant Gram-negative bacteria and to a great public health concern, considering that colistin is one of the last-resort antibiotics against multidrug-resistant deadly infections in clinical practice. Previous studies performed on livestock animals in Tunisia using culture-dependent methods highlighted the presence of colistin-resistant Gram-negative bacteria. In the present survey, DNA extracted from cloacal swabs from 195 broiler chickens from six farms in Tunisia was tested via molecular methods for the ten mobilized colistin resistance (mcr) genes known so far. Of the 195 animals tested, 81 (41.5%) were mcr-1 positive. All the farms tested were positive, with a prevalence ranging from 13% to 93%. These results confirm the spread of colistin resistance in livestock animals in Tunisia and suggest that the investigation of antibiotic resistance genes by culture-independent methods could be a useful means of conducting epidemiological studies on the spread of antimicrobial resistance.202337106971
5601140.9992Presence of Staphylococcus spp. carriers of the mecA gene in the nasal cavity of piglets in the nursery phase. The presence of Staphylococcus spp. resistant to methicillin in the nasal cavity of swine has been previously reported. Considering the possible occurrence of bacterial resistance and presence of resistance genes in intensive swine breeding and the known transmissibility and dispersion potential of such genes, this study aimed to investigate the prevalence of resistance to different antibiotics and the presence of the mecA resistance gene in Staphylococcus spp. from piglets recently housed in a nursery. For this, 60 nasal swabs were collected from piglets at the time of their housing in the nursery, and then Staphylococcus spp. were isolated and identified in coagulase-positive (CoPS) and coagulase-negative (CoNS) isolates. These isolates were subjected to the disk-diffusion test to evaluate the bacterial resistance profile and then subjected to molecular identification of Staphylococcus aureus and analyses of the mecA gene through polymerase chain reaction. Of the 60 samples collected, 60 Staphylococcus spp. were isolated, of which 38 (63.33%) were classified as CoNS and 22 (36.67%) as CoPS. Of these, ten (45.45%) were identified as Staphylococcus aureus. The resistance profile of these isolates showed high resistance to different antibiotics, with 100% of the isolates resistant to chloramphenicol, clindamycin, and erythromycin, 98.33% resistant to doxycycline, 95% resistant to oxacillin, and 85% resistant to cefoxitin. Regarding the mecA gene, 27 (45%) samples were positive for the presence of this gene, and three (11.11%) were phenotypically sensitive to oxacillin and cefoxitin. This finding highlights the importance of researching the phenotypic profile of resistance to different antimicrobials and resistance genes in the different phases of pig rearing to identify the real risk of these isolates from a One Health perspective. The present study revealed the presence of samples resistant to different antibiotics in recently weaned production animal that had not been markedly exposed to antimicrobials as growth promoters or even as prophylactics. This information highlights the need for more research on the possible sharing of bacteria between sows and piglets, the environmental pressure within production environments, and the exposure of handlers during their transport, especially considering the community, hospital, and political importance of the presence of circulating resistant strains.202336634542
5626150.9992Antimicrobial Resistance (AMR) of Bacteria Isolated from Dogs with Canine Parvovirus (CPV) Infection: The Need for a Rational Use of Antibiotics in Companion Animal Health. Canine parvovirus type 2 (CPV-2) represents a major viral threat to dogs. Considering the potential effects of pets on antimicrobial resistance, information on the CPV and associated bacterial co-infections is limited. The aim of this study was to analyze the antimicrobial susceptibility and multidrug-resistance profiles of bacterial species from tissue samples of dogs with canine parvovirus infection. A set of PCR assays and sequence analyses was used for the detection and the molecular characterization of the CPV strains and other enteric viruses. Bacterial isolation, the determination of antimicrobial susceptibility via the disk diffusion method, and the determination of the minimum inhibitory concentration were performed. The detection of β-lactamase genes and toxin genes for specific bacteria was also carried out. CPV infection was confirmed in 23 dogs. Forty-three bacterial strains were isolated and all showed phenotypic resistance. Seventeen multidrug-resistant bacteria and bacteria with high resistance to third- and fourth-generation cephalosporins and metronidazole were detected. Almost 50% of the isolated Enterobacteriaceae were positive for at least one β-lactamase gene, with the majority carrying more genes as well. The evidence for multi-resistant bacteria with the potential for intra- or cross-species transmission should be further considered in a One Health approach.202235203745
1627160.9992Screening of colistin-resistant bacteria in livestock animals from France. Colistin is frequently used as a growth factor or treatment against infectious bacterial diseases in animals. The Veterinary Division of the European Medicines Agency (EMA) restricted colistin use as a second-line treatment to reduce colistin resistance. In 2020, 282 faecal samples were collected from chickens, cattle, sheep, goats, and pigs in the south of France. In order to track the emergence of mobilized colistin resistant (mcr) genes in pigs, 111 samples were re-collected in 2021 and included pig faeces, food, and water from the same location. All samples were cultured in a selective Lucie Bardet Jean-Marc Rolain (LBJMR) medium and colonies were identified using MALDI-TOF mass spectrometry and then antibiotic susceptibility tests were performed. PCR and Sanger sequencing were performed to screen for the presence of mcr genes. The selective culture revealed the presence of 397 bacteria corresponding to 35 different bacterial species including Gram-negative and Gram-positive. Pigs had the highest prevalence of colistin-resistant bacteria with an abundance of intrinsically colistin-resistant bacteria and from these samples one strain harbouring both mcr-1 and mcr-3 has been isolated. The second collection allowed us to identify 304 bacteria and revealed the spread of mcr-1 and mcr-3 in pigs. In the other samples, naturally, colistin-resistant bacteria were more frequent, nevertheless the mcr-1 variant was the most abundant gene found in chicken, sheep, and goat samples and one cattle sample was positive for the mcr-3 gene. Animals are potential reservoir of colistin-resistant bacteria which varies from one animal to another. Interventions and alternative options are required to reduce the emergence of colistin resistance and to avoid zoonotic transmissions.202236414994
1946170.9992Using honey bee colonies to monitor phenotypic and genotypic resistance to colistin. Colistin is a polymyxin antimicrobic mainly used to treat infection caused by multi-drug resistant Gram-negative bacteria. Mechanisms of colistin resistance are linked to the mobile colistin resistance (mcr) genes, which are transferable within mobile plasmids. Currently, there is limited research on the environmental dissemination of these genes. The behavioural and morphological characteristics of Apis mellifera L. make honey bees effective environmental bioindicators for assessing the prevalence of antimicrobial-resistant bacteria. This study aims to evaluate the colistin phenotypic and genotypic resistance in environmental Gram-negative bacteria isolated from foraging honey bees, across a network of 33 colonies distributed across the Emilia-Romagna region in Italy. Phenotypic resistances were determined through a microdilution assay using the minimum inhibitory concentration (MIC) with dilutions ranging from 0.5 μg/ml to 256 μg/ml. Strains with MIC values gather than 2 μg/ml were classified as resistant. Also, the identification of the nine mcr genes was carried out using two separate multiplex PCR assays. The study found that 68.5% of isolates were resistant and the genus with the higher resistance rates observed in Enterobacter spp. (84.5%). At least one mcr gene was found in 137 strains (53.3%). The most detected gene was mcr5 (35.3%), which was the most frequently detected gene in the seven provinces, while the least observed was mcr4 (4.8%), detected only in two provinces. These results suggested the feasibility of detecting specific colistin resistance genes in environmentally spread bacteria and understanding their distribution at the environmental level, despite their restricted clinical use. In a One-Health approach, this capability enables valuable environmental monitoring, considering the significant role of colistin in the context of public health.202438944352
1968180.9992Multidrug-resistant pattern of food borne illness associated bacteria isolated from cockroaches in meal serving facilities, Jimma, Ethiopia. INTRODUCTION: An increase in the emergence and spread of multidrug-resistant (MDR) bacteria in recent years is becoming worrisome. Domestic cockroaches can play a significant role in the dissemination of such bacteria between the environment and human beings. This study aimed at determining anti-microbial resistance pattern of food borne illness associated bacteria identified from cockroaches trapped in restaurants and cafeterias. METHODS: Trapped cockroaches were picked with surgical gloves, sealed in sterile plastic bags and transported to the Microbiology laboratory. Standard microbiological techniques were used to isolate and identify bacteria. Anti-microbial susceptibility testing was done using Kirby Bauer diffusion technique. RESULT: A total of five species of food borne illness associated bacteria were detected. Majority (57.1%) of the bacteria were isolated from the gut of cockroaches. More than 89% of the isolates were multi drug resistance (MDR). MDR was higher on gram positive bacteria. S. aureus showed 53.3% resistance against oxacillin(MRSA) and 33.3% against vancomycin. CONCLUSION: A very high percentage of MDR bacteria was seen in this study. Most of the bacteria tested were isolated from the gut of cockroaches. Potential factors associated with cockroaches that contributed to this high MDR rate of the isolates should be investigated in future.201829977255
5599190.9992Antimicrobial susceptibility profiles of Staphylococcus spp. contaminating raw goat milk. BACKGROUND AND AIM: Antimicrobial resistance poses a major threat to global public health. Foodstuff of animal origin can serve as potential vehicles for the dissemination of antimicrobial-resistant bacteria and resistance genes to consumers. In view of the lack of knowledge about antimicrobial resistance in bacteria associated with goat milk, the aim of this study was to report species-level identification and antimicrobial susceptibility profiles of a large collection of Staphylococcus spp. isolates recovered from raw goat milk in Brazil. MATERIALS AND METHODS: A total of 434 Staphylococcus spp. isolates originated from 510 goat milk samples in Northeast Brazil were investigated. The isolates were obtained by conventional microbiological methods. Species identification and antimicrobial susceptibility testing were performed by means of a semi-automated system using a panel for biochemical tests and broth microdilution method for 19 antimicrobial drugs. RESULTS: Although Staphylococcus aureus (22.6%) accounted for the majority of the isolates, a total of 13 different non-aureus staphylococci spp. were identified. High resistance rates against erythromycin (40.8%), and the beta-lactams ampicillin (45.9%) and penicillin (42.9%) were observed among S. aureus isolates. The most significant findings were related to the resistance against quinupristin-dalfopristin, a drug of last resort used in human medicine to treat infections caused by vancomycin-resistant S. aureus and enterococci. CONCLUSION: The high diversity of Staphylococcus spp. showing phenotypic resistance against different antimicrobial drugs encourages further investigations on the real impact of these bacteria as reservoirs of antimicrobial resistance genes to consumers. Furthermore, the potential impact of technological processes, such as pasteurization, fermentation, and maturation, on the maintenance and dissemination of antimicrobial resistance among the microbial populations in milk and dairy products must also be investigated.202134220106