Surveillance of antimicrobial resistance in bacteria isolated from food animals to antimicrobial growth promoters and related therapeutic agents in Denmark. - Related Documents




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365601.0000Surveillance of antimicrobial resistance in bacteria isolated from food animals to antimicrobial growth promoters and related therapeutic agents in Denmark. This study was conducted to describe the occurrence of acquired resistance to antimicrobials used for growth promotion among bacteria isolated from swine, cattle and poultry in Denmark. Resistance to structurally related therapeutic agents was also examined. Three categories of bacteria were tested: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter, Salmonella, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). All antimicrobials used as growth promoters in Denmark and some structurally related therapeutic agents (in brackets) were included: Avilamycin, avoparcin (vancomycin), bacitracin, carbadox, flavomycin, monensin, olaquindox, salinomycin, spiramycin (erythromycin, lincomycin), tylosin (erythromycin, lincomycin), and virginiamycin (pristinamycin). Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Breakpoints for growth promoters were established by population distribution of the bacteria tested. A total of 2,372 bacterial isolates collected during October 1995 to September 1996 were included in the study. Acquired resistance to all currently used growth promoting antimicrobials was found. A frequent occurrence of resistance were observed to avilamycin, avoparcin, bacitracin, flavomycin, spiramycin, tylosin and virginiamycin, whereas resistance to carbadox, monensin, olaquindox and salinomycin was less frequent. The occurrence of resistance varied by animal origin and bacterial species. The highest levels of resistance was observed among enterococci, whereas less resistance was observed among zoonotic bacteria and bacteria pathogenic to animals. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed. The results show the present level of resistance to growth promoters in bacteria from food animals in Denmark. They will form the baseline for comparison with future prospective studies, thereby enabling the determination of trends over time.19989725794
556610.9998Resistance to antimicrobial agents used for animal therapy in pathogenic-, zoonotic- and indicator bacteria isolated from different food animals in Denmark: a baseline study for the Danish Integrated Antimicrobial Resistance Monitoring Programme (DANMAP). This study describes the establishment and first results of a continuous surveillance system of antimicrobial resistance among bacteria isolated from pigs, cattle and broilers in Denmark. The three categories of bacteria tested were: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter coli/jejuni, Salmonella enterica, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). A total of 3304 bacterial isolates collected from October 1995 through December 1996 were tested for susceptibility to all major classes of antimicrobial agents used for therapy in Denmark. Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Acquired resistance to all antimicrobials was found. The occurrence of resistance varied by animal origin and bacterial species. In general, resistance was observed more frequently among isolates from pigs than from cattle and broilers. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed, as is the occurrence of resistance in other countries. The results of this study show the present level of resistance to antimicrobial agents among a number of bacterial species isolated from food animals in Denmark. Thus, the baseline for comparison with future prospective studies has been established, enabling the determination of trends over time.19989744762
365720.9998A survey of antibiotic resistance in Micrococcaceae isolated from Italian dry fermented sausages. The transfer of bacteria that are resistant to antimicrobial agents or resistance genes from animals to humans via the food chain is increasingly a problem. Therefore, it is important to determine the species and the numbers of bacteria involved in this phenomenon. For this purpose, 148 strains of microstaphylococci were isolated from three types of Italian dry fermented sausages. Eight of 148 strains belonged to the genera Kocuria and Micrococcus. The remaining 140 strains belonged to 11 different species of the genus Staphylococcus. The species most frequently isolated was Staphylococcus xylosus, followed by Staphylococcus saprophyticus and Staphylococcus aureus. Antibiotic resistance levels differed among the species and depended on the strain origin. Microstaphylococci were generally susceptible to beta-lactams, but 12 strains were resistant to methicillin, 8 were resistant to oxacillin, and 9 were resistant to penicillin G. No resistance was observed for aminoglicosides and cephalosporines. Many strains were resistant to sulfonamide, colistin suphate, tetracyclin, and bacitracin. Two strains of S. aureus, four strains of S. xylosus, and one strain of Staphylococcus sciuri were able to grow in the presence of 8 microg of vancomycin per g, but all strains were susceptible to teicoplanin. Twenty-two microstaphylococci were resistant to at least five of the tested antibiotics. The multiresistant strain S. aureus 899 was unaffected by eight antibiotics, including vancomycin and methicillin, indicating that a more prudent use of antibiotics in animal husbandry and better hygienic conditions during production should be encouraged because they can play a major role in reducing the incidence of such multiresistant microorganisms and the possible spread of the genetic elements of their resistance.200312800992
559630.9997Enterotoxigenicity and Antibiotic Resistance of Coagulase-Negative Staphylococci Isolated from Raw Buffalo and Cow Milk. Staphylococcal food poisoning is considered to be one of the most common foodborne illnesses worldwide. Because milk is rich in nutrients and its neutral pH, it leads to the growth of various bacteria. To date, the correlation between enterotoxigenic potential in Staphylococcus species and antimicrobial resistance (AMR), using bioinformatics analysis in buffalo and cow raw milk and the possible health risks from these bacteria, has not been examined in Egypt. A total of 42 Staphylococcus isolates representing 12 coagulase-positive staphylococci (Staphylococcus aureus and Staphylococcus intermedius) and 30 coagulase-negative staphylococci (Staphylococcus capitis, Staphylococcus xylosus, Staphylococcus carnosus, Staphylococcus saccharolyticus, and Staphylococcus auricularis) were isolated. An assay of the antimicrobial resistance phenotypes indicated low resistance against vancomycin (9.5%). The blaZ gene was associated with penicillin G and methicillin resistance and not with sulbactam + ampicillin. The presence of the gene ermB presented the correlation with erythromycin resistance and tetK with tetracycline resistance (correlation index: 0.57 and 0.49, respectively), despite the absence of the same behavior for ermC and tetM, respectively. Interestingly, the gene mecA was not correlated with resistance to methicillin or any other β-lactam. Correlation showed that slime-producing isolates had more resistance to antibiotics than those of nonslime producers. The multiple correlations between antibiotic resistance phenotypes and resistance genes indicate a complex nature of resistance in Staphylococcus species. The antimicrobial resistance could potentially spread to the community and thus, the resistance of Staphylococcus species to various antibiotics does not depend only on the use of a single antimicrobial, but also extends to other unrelated classes of antimicrobials.202031750778
564740.9997Resistance of bacterial isolates from poultry products to therapeutic veterinary antibiotics. Bacterial isolates from poultry products were tested for their susceptibility to 10 antibiotics commonly used in the therapeutic treatment of poultry. Bacteria were isolated from fresh whole broiler carcasses or from cut-up meat samples (breast with or without skin, wings, and thighs) that were either fresh or stored at 4 or 13 degrees C (temperatures relevant to poultry-processing facilities). The Biolog system was used to identify isolates, and a broth dilution method was used to determine the antibiotic resistance properties of both these isolates and complementary cultures from the American Type Culture Collection. The antibiotics to which the most resistance was noted were penicillin G, sulfadimethoxine, and erythromycin; the antibiotic to which the least resistance was noted was enrofloxacin. Individual isolates exhibited resistances to as many as six antibiotics, with the most common resistance pattern involving the resistance of gram-negative bacteria to penicillin G, sulfadimethoxine, and erythromycin. Differences in resistance patterns were noted among 18 gram-positive and 7 gram-negative bacteria, and comparisons were made between species within the same genus. The data obtained in this study provide a useful reference for the species and resistance properties of bacteria found on various raw poultry products, either fresh or stored at temperatures and for times relevant to commercial processing, storage, and distribution. The results of this study show that resistance to antibiotics used for the therapeutic treatment of poultry occurs in bacteria in the processing environment.200312540187
590150.9997Identification and characterization of vancomycin-resistant Enterococcus species frequently isolated from laboratory mice. To determine the prevalence of drug resistant bacteria colonizing laboratory mice, we isolated and characterized vancomycin-resistant Enterococcus species (VRE) from commercially available mice. A total of 24 VRE isolates were obtained from 19 of 21 mouse strains supplied by 4 commercial breeding companies. Of these, 19 isolates of E. gallinarum and 5 isolates of E. casseliflavus possessing the vanC1 and vanC2/3 genes intrinsically, exhibited intermediate resistance to vancomycin respectively. In addition, these isolates also exhibited diverse resistant patterns to erythromycin, tetracycline, and ciprofloxacin, whereas the use of antibiotics had not been undertaken in mouse strains tested in this study. Although 6 virulence-associated genes (ace, asa, cylA, efaA, esp, and gelE) and secretion of gelatinase and hemolysin were not detected in all isolates, 23 of 24 isolates including the isolates of E. casselifalvus secreted ATP into culture supernatants. Since secretion of ATP by bacteria resident in the intestinal tract modulates the local immune responses, the prevalence of ATP-secreting VRE in mice therefore needs to be considered in animal experiments that alter the gut microflora by use of antibiotics.201425077759
550260.9997Short communication: Diversity of species and transmission of antimicrobial resistance among Staphylococcus spp. isolated from goat milk. The increasing production of goat milk and its derivatives is affected by the occurrence of intramammary infections, which are highly associated with the presence of Staphylococcus species, including some with zoonotic potential. Staphylococci in general can exchange mobile genetic elements, a process that may be facilitated by the isolate's capacity of forming biofilms. In this study we identified, to the species level, Staphylococcus isolated from goat milk samples by MALDI-TOF and confirmed the identification by sequencing housekeeping genes (rrs and tuf). Eight species were identified, more than half being either Staphylococcus epidermidis or Staphylococcus lugdunensis. The isolates were shown by pulsed-field gel electrophoresis to be genetically diverse between the studied herds. Resistance to ampicillin and penicillin was widespread, and 2 Staph. epidermidis isolates contained the methicillin-resistance gene mecA. Most of the isolates that were resistant to at least 1 of the 13 antimicrobials tested harbored plasmids, one of which was demonstrated to be conjugative, being transferred from a Staph. epidermidis to a Staphylococcus aureus strain. Biofilm formation was observed in almost every isolate, which may contribute to their capacity of exchanging antimicrobial resistance genes in addition to acting as a physical barrier to the access of drugs. Our results showed that antimicrobial resistance among goat staphylococci may be emerging in a process facilitated by the exchange of mobile genetic elements between the bacteria and the establishment of biofilms, which calls for careful monitoring and more effective control therapies.201930928272
599470.9997Characterization of Erythromycin and Tetracycline Resistance in Lactobacillus fermentum Strains. Lactobacillus fermentum colonizing gastrointestinal and urogenital tracts of humans and animals is widely used in manufacturing of fermented products and as probiotics. These bacteria may function as vehicles of antibiotic resistance genes, which can be transferred to pathogenic bacteria. Therefore, monitoring and control of transmissible antibiotic resistance determinants in these microorganisms is necessary to approve their safety status. The aim of this study was to characterize erythromycin and tetracycline resistance of L. fermentum isolates and to estimate the potential transfer of resistance genes from lactobacilli to the other Gram-positive and Gram-negative bacteria. Among six L. fermentum strains isolated from human feces and commercial dairy products, five strains demonstrated phenotypic resistance to tetracycline. PCR screening for antibiotic resistance determinants revealed plasmid-located tetracycline resistance genes tet(K) and tet(M) in all strains and erythromycin resistance genes erm(B) in the chromosome of L. fermentum 5-1 and erm(C) in the plasmid of L. fermentum 3-4. All tested lactobacilli lacked conjugative transposon Tn916 and were not able to transfer tetracycline resistance genes to Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, Acinetobacter baumannii, Citrobacter freundii, and Escherichia coli by filter mating. Staphylococcus haemolyticus did not accept erythromycin resistance genes from corresponding Lactobacillus strains. Thus, in the present study, L. fermentum was not implicated in the spread of erythromycin and tetracycline resistance, but still these strains pose the threat to the environment and human health because they harbored erythromycin and tetracycline resistance genes in their plasmids and therefore should not be used in foods and probiotics.201830534155
559780.9997Prevalence 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
282290.9997Antimicrobial resistance of bacterial flora associated with bovine products in South Africa. The administration of subtherapeutic doses of antibiotics to livestock introduces selective pressures that may lead to the emergence and dissemination of resistant bacteria. This study determined the antibiotic-resistance spectra of the microbial flora found on freshly slaughtered and retail beef and in unpasteurized and pasteurized packaged milk. Staphylococci, Enterobacteriaeae, and isolates from total aerobic plate counts were tested for resistance to vancomycin, streptomycin, methicillin, tetracycline, and gentamicin using the disc diffusion susceptibility test and resistance to penicillin was determined by using oxacillin. A larger proportion of resistance to most antibiotics, except for vancomycin, was displayed by isolates from abattoir samples. The incidence of multiple antibiotic resistance (MAR) pathogenic bacteria is also higher in the abattoir. Resistance genes lost because of lack of selective pressure or resistant flora being replaced by more sensitive flora during processing is the reason for the lower incidence of MAR pathogenic bacteria among retail samples. These resistant bacteria can be transferred to humans through the consumption of rare or raw beef and unpasteurized milk, thus rendering the resultant food-related infections difficult to treat. The present findings clearly demonstrate that antibiotic-resistant bacteria in beef and milk pose a serious problem in South Africa.199910382649
5646100.9997Dispersion and persistence of antimicrobial resistance genes among Staphylococcus spp. and Mammaliicoccus spp. isolated along a swine manure treatment plant. Staphylococcus spp. and Mammaliicoccus spp. colonize the skin and mucosa of humans and other animals and are responsible for several opportunistic infections. Staphylococci antibiotic resistance may be present in the environment due to the spread of treated and untreated manure from the livestock industry due to antibiotic use to disease control or growth promoter. In this work, we analyzed the species distribution and antimicrobial susceptibility of Staphylococcus and Mammaliicoccus species along different sites of a swine manure treatment plant from Southeastern Brazil. Bacterial colonies were obtained on mannitol salt agar, selected after catalase test and Gram staining, and finally identified by mass spectrometry and sequencing of the tuf gene. According to the results, S.cohnii and S. simulans were the most prevalent species. Antibiotic resistance test revealed that several strains were resistant to multiple drugs, with high levels of chloramphenicol resistance (98%), followed by erythromycin (79%), tetracycline (73%), gentamicin (46%), ciprofloxacin (42%), cefoxitin (18%), sulfamethoxazole + trimethoprim (12%), and linezolid (4%). In addition, gene detection by PCR showed that all strains carried at least 2 resistance genes and one of them carried all 11 genes investigated. Using the GTG(5)-PCR approach, a high genetic similarity was observed between some strains that were isolated from different points of the treatment plant. Although some were seemingly identical, differences in their resistance phenotype and genotype suggest horizontal gene transfer. The presence of resistant bacteria and resistance genes along the treatment system highlights the potential risk of contamination by people in direct contact with these animals and the soil since the effluent is used as a biofertilizer in the surrounding environment.202336515883
6065110.9997Screening for enterocins and detection of hemolysin and vancomycin resistance in enterococci of different origins. The inhibitory activity of 122 out of 426 Enterococcus strains of geographically widespread origin and from different sources (food and feed, animal isolates, clinical and nonclinical human isolates) was tested against a wide range of indicator bacteria. Seventy-two strains, mainly belonging to the species Enterococcus faecium and Enterococcus faecalis were bacteriocinogenic. A remarkable variation of inhibitory spectra occurred among the strains tested, including inhibition of, for instance, only closely related enterococci, other lactic acid bacteria (LAB), food spoilage and pathogenic bacteria. No correlation could be found between the origin of the strains and the type of inhibitory spectrum, although a clustering of human isolates from both fecal and clinical origin was observed in the group of strains inhibiting lactic acid bacteria, Listeria, and either Staphylococcus or Clostridium. No relationship could be established between the presence of enterocin structural genes and the origin of the strain either, and hence no correlation seemed to exist between the presence of known enterocin genes and the activity spectra of these enterococci. The structural gene of enterocin A was widely distributed among E. faecium strains, whereas that of enterocin B only occurred in the presence of enterocin A. The vancomycin resistance phenotype as well as the presence of vancomycin resistance genes was also investigated. The vanA gene only occurred among E. faecium strains. The incidence of beta-hemolysis was not restricted to E. faecalis strains, but among the E. faecium strains the structural genes of cytolysin were not detected. beta-Hemolysis occurred in strains both from food and nonfood origin. It has been concluded that bacteriocin-producing E. faecium strains lacking hemolytic activity and not carrying cytolysin nor vancomycin resistance genes may be useful as starter cultures, cocultures, or probiotics.200312810293
5546120.9997Antibiotic resistance and Caco-2 cell invasion of Pseudomonas aeruginosa isolates from farm environments and retail products. The potential pathogenicity of Pseudomonas aeruginosa isolates from food animals, retail meat products, and food processing environments was evaluated by determining their antibiotic resistance profiles and invasiveness into human intestinal Caco-2 cell. In general, the genomically diversified isolates of P. aeruginosa were resistant to beta-lactams (ampicillin, amoxicillin-clavulanic acid, cefoxitin, ceftiofur, and cephalothin), chloramphenicol, tetracycline, kanamycin, nalidixic acid, and sulfamethoxazole-trimethoprim. Acquisition of any other antibiotic resistance genes, such as class 1 integrons and other beta-lactamase genes, was not found in the tested isolates. The expression of OprM membrane protein, which is associated with a multidrug efflux system, played a major role in their antibiotic resistance. Single mutation in the GyrA to confer resistance to nalidixic acids was also found in the tested isolates, indicating that these factors could synergistically affect the resistance of the P. aeruginosa isolates. The number of bacteria invading into the Caco-2 cells was 2.5 log(10) CFU/ml on average. Therefore, the public health concern of P. aeruginosa could be relevant since its occurrence in food animals could cross contaminate the retail meat products during food handling and processing.200717289197
4592130.9997The Genetic Diversity and Antimicrobial Resistance of Pyogenic Pathogens Isolated from Porcine Lymph Nodes. According to the Food and Agriculture Organization of the United Nations, pork remains the most consumed meat in the world. Consequently, it is very important to ensure that it is of the highest microbiological quality. Many of the pathogens that cause lymph node lesions in pigs are zoonotic agents, and the most commonly isolated bacteria are Mycobacterium spp., Streptococcus spp., Staphylococcus aureus and Rhodococcus equi (synonymous with Prescottella equi). The prevention and treatment of zoonotic infections caused by these bacteria are mainly based on antimicrobials. However, an overuse of antimicrobials contributes to the emergence and high prevalence of antimicrobial-resistant strains, which are becoming a serious challenge in many countries. The aim of this study was to evaluate the genetic diversity and antimicrobial resistance of the Streptococcus spp. (n = 48), S. aureus (n = 5) and R. equi (n = 17) strains isolated from swine lymph nodes with and without lesions. All isolates of S. dysgalactiae, S. aureus and R. equi were subjected to PFGE analysis, which showed the genetic relatedness of the tested bacteria in the studied pig populations. Additionally, selected tetracycline and macrolide resistance genes in the streptococcal strains were also studied. The results obtained in the present study provide valuable data on the prevalence, diversity, and antimicrobial resistance of the studied bacteria. Numerous isolated bacterial Streptococcus spp. strains presented resistance to doxycycline, and almost half of them carried tetracycline resistance genes. In addition, R. equi and S. aureus bacteria presented a high level of resistance to beta-lactam antibiotics and to cefotaxime, respectively.202337370345
5910140.9996Antimicrobial Susceptibility Testing and Tentative Epidemiological Cutoff Values for Five Bacillus Species Relevant for Use as Animal Feed Additives or for Plant Protection. Bacillus megaterium (n = 29), Bacillus velezensis (n = 26), Bacillus amyloliquefaciens (n = 6), Bacillus paralicheniformis (n = 28), and Bacillus licheniformis (n = 35) strains from different sources, origins, and time periods were tested for the MICs for nine antimicrobial agents by the CLSI-recommended method (Mueller-Hinton broth, 35°C, for 18 to 20 h), as well as with a modified CLSI method (Iso-Sensitest [IST] broth, 37°C [35°C for B. megaterium], 24 h). This allows a proposal of species-specific epidemiological cutoff values (ECOFFs) for the interpretation of antimicrobial resistance in these species. MICs determined by the modified CLSI method were 2- to 16-fold higher than with the CLSI-recommended method for several antimicrobials. The MIC distributions differed between species for five of the nine antimicrobials. Consequently, use of the modified CLSI method and interpretation of resistance by use of species-specific ECOFFs is recommended. The genome sequences of all strains were determined and used for screening for resistance genes against the ResFinder database and for multilocus sequence typing. A putative chloramphenicol acetyltransferase (cat) gene was found in one B. megaterium strain with an elevated chloramphenicol MIC compared to the other B. megaterium strains. In B. velezensis and B. amyloliquefaciens, a putative tetracycline efflux gene, tet(L), was found in all strains (n = 27) with reduced tetracycline susceptibility but was absent in susceptible strains. All B. paralicheniformis and 23% of B. licheniformis strains had elevated MICs for erythromycin and harbored ermD The presence of these resistance genes follows taxonomy suggesting they may be intrinsic rather than horizontally acquired. Reduced susceptibility to chloramphenicol, streptomycin, and clindamycin could not be explained in all species.IMPORTANCE When commercializing bacterial strains, like Bacillus spp., for feed applications or plant bioprotection, it is required that the strains are free of acquired antimicrobial resistance genes that could potentially spread to pathogenic bacteria, thereby adding to the pool of resistance genes that may cause treatment failures in humans or animals. Conversely, if antimicrobial resistance is intrinsic to a bacterial species, the risk of spreading horizontally to other bacteria is considered very low. Reliable susceptibility test methods and interpretation criteria at the species level are needed to accurately assess antimicrobial resistance levels. In the present study, tentative ECOFFs for five Bacillus species were determined, and the results showed that the variation in MICs followed the respective species. Moreover, putative resistance genes, which were detected by whole-genome sequencing and suggested to be intrinsic rather that acquired, could explain the resistance phenotypes in most cases.201830030233
5498150.9996The prevalence of multidrug resistance in Staphylococcus hominis isolated from clinical materials. The treatment of infections caused by Staphylococcus hominis remains a challenge, mainly due to the increasing resistance of these bacteria to antibiotics. The aim of the study was to determine antibiotic resistance in 62 strains S. hominis isolated from clinical materials, and to identify the molecular basis of resistance to antibiotics. Forty-six strains were both methicillin-resistant and harbored the mecA gene. Twenty-three of these strains had mec complex A and ccr complex AB1. Such a combination of the mec and ccr complexes does not correspond to any cassettes that have been demonstrated so far. However, over 80% of the tested strains were multidrug-resistant, of which as many as 12 were resistant to at least seven antibiotics. More than a half of strains harbored the tetK, acc(6')-Ie aph(2''), and ant(4')-I genes. erm(C) was the most common resistant gene to antibiotics from the MLS group. Two strains had as many as five antibiotic resistance genes from the tested groups (erm(C), msr(A), msr(B), mph(C), lnu(A)). The presence of the vga gene encoding resistance to streptogramins A was detected in one strain. All of strains were sensitive to vancomycin. However, 11 of them had reduced sensitivity to this antibiotic and eight of them were characterized by a heterogeneous resistance profile to this antibiotic. Our results clearly shows increasing threat of S. hominis caused by their multi-resistance. Moreover, these bacteria can constitute a reservoir of resistance genes for more pathogenic bacteria.202539747570
5565160.9996Vancomycin resistance and virulence genes evaluation in Enterococci isolated from pork and wild boar meat. Enterococci are considered valuable sentinel Gram-positive bacteria for monitoring vancomycin antibiotic resistance due to their widespread presence and characteristics. The use of antimicrobials in farming animals has a role in the increasing of Antimicrobial Resistance (AMR) and the anthropogenic transformation of the landscape has forced wildlife into greater contact with humans and their livestock. The transmission of resistant bacteria by their meat products is a significant contributor to AMR development. The present study aimed to assess the prevalence of vancomycin resistant Enterococci spp. In antimicrobial-treated farmed pigs meat and in antimicrobial-free wild boars meat. A total of 341 Enterococci were isolated from 598 pork meat samples (57 %) and 173 Enterococci were isolated from 404 wild boar meat samples (42.8 %). Data found showed that low-resistance was detected more in wild boars meat Enterococci (52.6 %) than in pork meat once (48.4 %). However, the prevalence of resistance genes was at low level (33.9 % in pork meat Enterococci and 4.4 % in wild boar meat ones) and the only gene found was vanC1/C2, related to intrinsic AMR. Normally, Enterococci persist in the normal intestinal flora of animals including humans. However, the presence of resistance genes was frequently linked to the detection of pathogenic genes, mostly gelE in pork meat isolates and asa1 in wild boars meat isolates. Pathogenic bacteria can cause severe infections in human that can become more risky if associated to the presence of AMR. Pathogenic bacteria were characterized and a high presence of E. gallinarum and E. casseliflavus was found. Given the growing interest in wild game meat consumption the monitoring of AMR in these matrices is essential. Further surveillance studies are needed to fully evaluate the emergence and spread of vancomycin-resistant Enterococci (VRE) and pathogenic Enterococci from animal-derived food to humans, including the role of wildlife in this phenomenon. Giving the higher interest in wild animals meat consumption, it is important to better evaluate the spread of AMR phenomenon in the future and intensify hygienic control of wild animals derived food.202439104496
5500170.9996Whole genome sequence analyses-based assessment of virulence potential and antimicrobial susceptibilities and resistance of Enterococcus faecium strains isolated from commercial swine and cattle probiotic products. Enterococcus faecium is one of the more commonly used bacterial species as a probiotic in animals. The organism, a common inhabitant of the gut of animals and humans, is a major nosocomial pathogen responsible for a variety infections in humans and sporadic infections in animals. In swine and cattle, E. faecium-based probiotic products are used for growth promotion and gut functional and health benefits. The objective of this study was to utilize whole genome sequence-based analysis to assess virulence potential, detect antimicrobial resistance genes, and analyze phylogenetic relationships of E. faecium strains from commercial swine and cattle probiotics. Genomic DNA extracted from E. faecium strains, isolated from commercial probiotic products of swine (n = 9) and cattle (n = 13), were sequenced in an Illumina MiSeq platform and analyzed. Seven of the nine swine strains and seven of the 13 cattle strains were identified as Enterococcus lactis, and not as E. faecium. None of the 22 probiotic strains carried major virulence genes required to initiate infections, but many carried genes involved in adhesion to host cells, which may benefit the probiotic strains to colonize and persist in the gut. Strains also carried genes encoding resistance to a few medically important antibiotics, which included aminoglycosides [aac(6')-Ii, aph(3')-III, ant(6)-Ia], macrolide, lincosamide and streptogramin B (msrC), tetracyclines [tet(L) and tet(M)], and phenicols [cat-(pc194)]. The comparison of the genotypic to phentypic AMR data showed presence of both related and unrelated genes in the probiotic strains. Swine and cattle probiotic E. faecium strains belonged to diverse sequence types. Phylogenetic analysis of the probiotic strains, and strains of human (n = 29), swine (n = 4), and cattle (n = 4) origin, downloaded from GenBank, indicated close clustering of strains belonging to the same species and source, but a few swine and cattle probiotic strains clustered closely with other cattle and human fecal strains. In conclusion, the absence of major virulence genes characteristic of the clinical E. faecium strains suggests that these probiotic strains are unlikely to initiate opportunistic infection. However, the carriage of AMR genes to medically important antibiotics and close clustering of the probiotic strains with other human and cattle fecal strains suggests that probiotic strains may pose risk to serve as a source of transmitting AMR genes to other gut bacteria.202235150575
5908180.9996Evaluation of Tetracycline Resistance and Determination of the Tentative Microbiological Cutoff Values in Lactic Acid Bacterial Species. Lactic acid bacteria (LAB) are widely used as probiotics in the food industry owing to their beneficial effects on human health. However, numerous antibiotic resistance genes have been found in LAB strains, especially tetracycline resistance genes. Notably, the potential transferability of these genes poses safety risks. To comprehensively evaluate tetracycline resistance in LAB, we determined the tetracycline susceptibility patterns of 478 LAB strains belonging to four genera and eight species. By comparing phenotypes with genotypes based on genome-wide annotations, five tetracycline resistance genes, tet(M), tet(W/N/W), tet(L), tet(S), and tet(45), were detected in LAB. Multiple LAB strains without tetracycline resistance genes were found to be resistant to tetracycline at the currently recommended cutoff values. Thus, based on the minimum inhibitory concentrations of tetracycline for these LAB strains, the species-specific microbiological cutoff values for Lactobacillus (para)gasseri, Lactobacillus johnsonii, and Lactobacillus crispatus to tetracycline were first developed using the Turnidge, Kronvall, and eyeball methods. The cutoff values for Lactiplantibacillus plantarum were re-established and could be used to better distinguish susceptible strains from strains with acquired resistance. Finally, we verified that these five genes play a role in tetracycline resistance and found that tet(M) and tet(W/N/W) are the most widely distributed tetracycline resistance genes in LAB.202134683449
5598190.9996Antibiotic Resistance in Lactic Acid Bacteria from Dairy Products in Northern Italy. Background: The spread of antibiotic resistance genes (ARGs) from the food chain is a significant public health concern. Dairy products from raw milk containing lactic acid bacteria (LAB) resistant to antimicrobials may serve as vectors for the transfer of resistance to commensal or potentially pathogenic bacteria in the human gut. Detecting ARGs in dairy products and milk is, therefore, crucial and could aid in the development of strategies to mitigate resistance dissemination through the food chain. Objectives: This study aimed to determine the presence of ARGs and assess the antibiotic susceptibility of LAB strains isolated from dairy products made from raw milk. Methods: Fifty-four LAB strains were isolated from 41 dairy samples and were tested for antimicrobial susceptibility using broth microdilution to determine Minimal Inhibitory Concentration (MIC). Moreover, the presence of resistance genes related to tetracyclines, beta-lactams, quinolones, and erythromycin was examined using six multiplex PCR assays. Results: Lactobacillus spp. and Leuconostoc spp. strains exhibited a high level of resistance to vancomycin (93-100%). Low-level resistance (4.2-20%) was observed in Lactococcus spp. and Lactobacillus spp. strains against tetracycline. Additionally, Lactococcus spp. strains showed resistance to trimethoprim/sulfamethoxazole, erythromycin, and clindamycin. Twenty-two out of 54 LAB strains (40.7%) carried at least one antibiotic resistance gene, and five of these were multidrug-resistant. Genes associated with acquired resistance to tetracycline were commonly detected, with tetK being the most frequent determinant. Conclusions: This study demonstrated that LABs in dairy products can act as reservoirs for ARGs, potentially contributing to the horizontal transfer of resistance within microbial communities in food and consumers. These findings highlight the need for the ongoing surveillance of antibiotic resistance in LAB and the implementation of control measures to minimize the dissemination of resistance through dairy products.202540298519