Susceptibility of Lactobacillaceae Strains to Aminoglycoside Antibiotics in the Light of EFSA Guidelines. - Related Documents




#
Rank
Similarity
Title + Abs.
Year
PMID
012345
591401.0000Susceptibility of Lactobacillaceae Strains to Aminoglycoside Antibiotics in the Light of EFSA Guidelines. Lactobacillaceae is a large family of bacteria from which probiotic strains often originate. Microorganisms used as feed additives in the EU must meet a number of formal criteria, some of which concern antimicrobial susceptibility. In this study, we determined the susceptibility of 19 reference strains and 121 wild-type strains of Lactobacillaceae to aminoglycoside antibiotics using the broth microdilution method based on the ISO 10932:2010/IDF 223:2010 standard. Strains were categorized as resistant or susceptible according to European Food Safety Authority (EFSA) guidelines. Resistance genes were detected by whole genome sequence (WGS) analysis or by PCR. The MICs read after 48 h of incubation showed that 36.8% of reference strains were resistant to kanamycin, 26.3% to streptomycin, and 5.3% to gentamicin, with no aminoglycoside resistance genes detected in any genome. As many as 93.2% of field isolates of Ligilactobacillus salivarius, 85% of Ligilactobacillus agilis, and 58.8% of Lactiplantibacillus plantarum were classified as resistant to kanamycin, with the aac(6)-Ie-aph(2)-Ia gene detected only in two isolates. In six of 12 streptomycin-resistant strains, the ant(6)-Ia gene was identified, which usually coexisted with the spw gene. Three isolates with high neomycin MICs harbored the ant(4')-Ia gene. In Lactobacillus gallinarum strain LMG 9435, characterized by streptomycin MIC value > 1024 µg/mL, a potential resistance-causing mutation in the rpsL gene (Lys56 → Arg) was detected. The results of the study indicate that some genera of Lactobacillaceae, in particular L. salivarius and L. agilis, exhibit natural resistance to aminoglycoside antibiotics, mainly kanamycin. Therefore, there is a need to update the EFSA guidelines on antimicrobial susceptibility testing of Lactobacillaceae, so that strains lacking resistance genes and/or chromosomal mutations are not considered to be resistant.202540430158
238710.9998Phenotypic and genotypic antimicrobial resistance profiles of fecal lactobacilli from domesticated pigeons in Poland. Lactobacillus species play an important role in the host and although they are non-pathogenic, they could act as reservoirs for antibiotic resistance genes, with the potential risk of transfer to other bacteria inhabiting the gastrointestinal tract. The aim of this study was to identify Lactobacillus species derived from feces of domesticated pigeons and to characterize their phenotypic and genotypic antimicrobial resistance (AMR) profiles. A total of 57 Lactobacillus isolates were classified into six species using the MALDI-TOF technique and 16S rDNA restriction analysis. Strains of L. ingluviei (31%), L. salivarius (28%) and L. agilis (23%) were the dominant species isolated. Determination of antimicrobial susceptibility by the microdilution broth method showed widespread resistance to kanamycin (89%), tetracycline (84%), streptomycin (63%), and enrofloxacin (37%). Less than 30% of the isolates were resistant to erythromycin, lincosamides, gentamycin, chloramphenicol and vancomycin. Over half (51%) of the lactobacilli were classified as multidrug resistant. Tet genes were detected in 79% of isolates; the lnuA, cat, ermB, ermC, ant(6)-Ia, ant(4')-Ia, and int-Tn genes were found at a lower frequency. Sequence analysis of the quinolone resistance-determining region (QRDR)of the gyrA gene showed that fluoroquinolone resistance in lactobacilli was the result of a mutation that lead to a change in the amino acid sequence (Ser83→Tyr/Leu/Phe). Domesticated pigeons could be a reservoir for AMR Lactobacillus strains and AMR genes.202032781109
238920.9997Antibiotic Resistance of LACTOBACILLUS Strains. The study provides phenotypic and molecular analyses of the antibiotic resistance in 20 Lactobacillus strains including 11 strains newly isolated from fermented plant material. According to the results of disc diffusion method, 90% of tested lactobacilli demonstrated sensitivity to clindamycin and 95% of strains were susceptible to tetracycline, erythromycin, and rifampicin. Ampicillin and chloramphenicol were found to inhibit all bacteria used in this study. The vast majority of tested strains revealed phenotypic resistance to vancomycin, ciprofloxacin, and aminoglycosides. Most of Lactobacillus strains showed high minimum inhibitory concentrations (MICs) of cefotaxime, ceftriaxone, and cefazolin and therefore were considered resistant to cephalosporins. All the strains exhibited multidrug resistance. The occurrence of resistance genes was associated with phenotypic resistance, with the exception of phenotypically susceptible strains that contained genes for tetracycline (tetK, tetL) and erythromycin (ermB, mefA) resistance. The vanX gene for vancomycin resistance was among the most frequently identified among the lactobacilli (75% of strains), but the occurrence of the parC gene for ciprofloxacin resistance was sporadic (20% of strains). Our results mainly evidence the intrinsic nature of the resistance to aminoglycosides in lactobacilli, though genes for enzymatic modification of streptomycin aadA and aadE were found in 20% of tested strains. The occurrence of extended spectrum beta-lactamases (ESBL) was unknown in Lactobacillus, but our results revealed the blaTEM gene in 80% of strains, whereas blaSHV and blaOXA-1 genes were less frequent (20% and 15% of strains, respectively).201931555856
599730.9997Resistance of potential probiotic lactic acid bacteria and bifidobacteria of African and European origin to antimicrobials: determination and transferability of the resistance genes to other bacteria. Probiotic bacteria and starter cultures of Lactobacillus, Weissella and Bifidobacterium of African and European origins were studied and compared for their susceptibility to antimicrobials. The study included, for all isolates, determination of MICs (Minimal Inhibitory Concentration) for 24 antimicrobials, detection of resistance genes by PCR reactions using specific primers and sequencing of positive amplicons. The ability of Lb. reuteri from Africa to transfer the erythromycin resistance gene erm(B) to closely related bacteria was investigated by conjugation. Variations were observed and high levels of intrinsic resistance were found among the tested species. Positive amplicons were obtained for resistance genes encoding aminoglycoside (aph(3')-III, aadA, aadE) and tetracycline (tet(S)) from isolates from Europe and macrolide (erm(B)) from an isolate from Africa. However, only the erm(B) gene found in Lb. reuteri L4: 12002 from Africa contained a homologous sequence to previously published sequences. This gene could be transferred in vitro to enterococci. Higher prevalence of phenotypic resistance for aminoglycoside was found in isolates from Europe.200818063151
238840.9997Antibiotic resistance in lactic acid bacteria isolated from some pharmaceutical and dairy products. A total of 244 lactic acid bacteria (LAB) strains were isolated from 180 dairy and pharmaceutical products that were collected from different areas in Minia governorate, Egypt. LAB were identified phenotypically on basis of morphological, physiological and biochemical characteristics. Lactobacillus isolates were further confirmed using PCR-based assay. By combination of phenotypic with molecular identification Lactobacillus spp. were found to be the dominant genus (138, 76.7%) followed by Streptococcus spp. (65, 36.1%) and Lactococcus spp. (27, 15%). Some contaminant organisms such as (Staphylococcus spp., Escherichia coli, Salmonella spp., mould and yeast) were isolated from the collected dairy samples but pharmaceutical products were free of such contaminants. Susceptibility of LAB isolates to antibiotics representing all major classes was tested by agar dilution method. Generally, LAB were highly susceptible to Beta-lactams except penicillin. Lactobacilli were resistant to vancomycin, however lactococci and streptococci proved to be very susceptible. Most strains were susceptible to tetracycline and showed a wide range of streptomycin MICs. The MICs of erythromycin and clindamycin for most of the LAB were within the normal range of susceptibility. Sixteen Lactobacillus, 8 Lactococcus and 8 Streptococcus isolates including all tetracycline and/or erythromycin resistant strains were tested for the presence of tetracycline and/or erythromycin resistant genes [tet(M) and/or erm(B)]. PCR assays shows that some resistant strains harbor tet(M) and/or erm(B) resistance genes.201424948910
593650.9997Antibiotic Resistance Characterization and Molecular Characteristics of Enterococcus Species Isolated from Combination Probiotic Preparations in China. Enterococci can act as reservoirs for antibiotic-resistant genes that are potentially at risk of being transferred to other bacteria that inhabit in the gastrointestinal tract. The aim of this study was to determine the phenotypic and molecular characteristics of antibiotic-resistant enterococci isolated from probiotic preparations. In total, we isolated 15 suspected Enterococcus species from 5 compound probiotics, which were identified by 16S rDNA as 12 Enterococcus faecium and 3 Enterococcus faecalis. Determination of antimicrobial susceptibility by the microdilution broth method showed widespread resistance to sulfamethoxazole (100%), norfloxacin (99.3%), azithromycin (99.3%), gentamicin (86.7%), and chloramphenicol (20%). Whole genome sequencing of five resistant strains revealed that all had circular DNA chromosomes and that E. faecium J-1-A to J-4-A contained a plasmid, while E. faecalis J-5-A did not. The results of the resistance gene analysis revealed that each strain contained approximately 30 resistance genes, with the antibiotic resistance genes and the multidrug resistance efflux pump genes mdtG, lmrC, and lmrD detected in all strains. The chloramphenicol resistance genes ykkC and ykkD were first identified in E. faecalis. And there were 21, 19, 21, 21, and 29 virulence factors involved in strains, respectively. Further analysis of the gene islands (GIs) revealed that each strain contained more than 10 GIs. The above results confirm the existence of hidden dangers in the safety of probiotics and remind us to carefully select probiotic preparations containing enterococcal strains to avoid the potential spread of resistance and pathogenicity.202437824752
239960.9997Ready-to-eat dairy products as a source of multidrug-resistant Enterococcus strains: Phenotypic and genotypic characteristics. The enterococci are ubiquitous bacteria able to colonize the human and animal gastrointestinal tracts and fresh and fermented food products. Their highly plastic genome allows Enterococcus spp. to gain resistance to multiple antibiotics, making infections with these organisms difficult to treat. Food-borne enterococci could be carriers of antibiotic resistance determinants. The goal of this work was to study the characteristics of Enterococcus spp. in fermented milk products from Poland and their antibiotic resistance gene profiles. A total of 189 strains were isolated from 182 dairy products out of 320 samples tested. The predominant species were Enterococcus faecium (53.4%) and Enterococcus faecalis (34.4%). Isolates were resistant to streptomycin (29.1%), erythromycin (14.3%), tetracycline (11.6%), rifampicin (8.7%), and tigecycline (8.1%). We also detected 2 vancomycin-resistant and 3 linezolid-resistant strains; however, no vanA or vanB genes were identified. A total of 57 high-level aminoglycoside resistance strains (30.2%) were identified, most of which have the ant(6')-Ia gene, followed by the aac(6')-Ie-aph(2″)-Ia and aph(3″)-IIIa genes. Resistance to tetracycline was most often conferred by tetM and tetL genes. Macrolide resistance was most frequently encoded by ermB and ermA genes. Conjugative mobile genetic element (transposon Tn916-Tn1545) was identified in 15.3% of the strains, including 96.3% of strains harboring the tetM gene. This study found that enterococci are widely present in retail ready-to-eat dairy products in Poland. Many isolated strains are antibiotic resistant and carry transferable resistance genes, which represent a potential source of transmission of multidrug-resistant bacteria to humans.202032197843
268570.9997Prevalence and genomic characterization of the Bacillus cereus group strains contamination in food products in Southern China. The Bacillus cereus group, as one of the important opportunistic foodborne pathogens, is considered a risk to public health due to foodborne diseases and an important cause of economic losses to food industries. This study aimed to gain essential information on the prevalence, phenotype, and genotype of B. cereus group strains isolated from various food products in China. A total of 890 strains of B. cereus group bacteria from 1181 food samples from 2020 to 2023 were identified using the standardized detection method. These strains were found to be prevalent in various food types, with the highest contamination rates observed in cereal flour (55.8 %) and wheat/rice noodles (45.7 %). The tested strains exhibited high resistance rates against penicillin (98.5 %) and ampicillin (98.9 %). Strains isolated from cereal flour had the highest rate of meropenem resistance (7.8 %), while strains from sausages were most resistant to vancomycin (16.8 %). A total of 234 out of the 891 B. cereus group strains were randomly selected for WGS analysis, 18.4 % of which displayed multidrug resistance. The species identification by WGS analysis revealed the presence of 10 distinct species within the B. cereus group, with B. cereus species being the most prevalent. The highest level of species diversity was observed in sausages. Notably, B. anthracis strains lacking the anthrax toxin genes were detected in flour-based food products and sausages. A total of 20 antibiotic resistance genes have been identified, with β-lactam resistance genes (bla1, bla2, BcI, BcII, and bla(TEM-116)) being the most common. The B. tropicus strains exhibit the highest average number of virulence genes (23.4). The diarrheal virulence genes nheABC, hblACD, and cytK were found in numerous strains. Only 4 of the 234 (1.7 %) sequenced strains contain the ces gene cluster linked to emetic symptoms. These data offer valuable insights for public health policymakers on addressing foodborne B. cereus group infections and ensuring food safety.202438354793
539780.9997Antimicrobial Resistance of Seventy Lactic Acid Bacteria Isolated from Commercial Probiotics in Korea. In this study, lactic acid bacteria were isolated from 21 top-selling probiotic products on Korean market and their antimicrobial resistance were analyzed. A total 152 strains were claimed to be contained in these products and 70 isolates belonging to three genera (Bifidobacterium, Lactobacillus, and Lactococcus) were obtained from these products. RAPD-PCR showed diversity among isolates of the same species except for two isolates of Lacticaibacillus rhamnosus from two different products. The agar dilution method and the broth dilution method produced different MICs for several antimicrobials. With the agar dilution method, five isolates (three isolates of Bifidobacterium animalis subsp. lactis, one isolate of B. breve, one isolate of B. longum) were susceptible to all nine antimicrobials and 15 isolates were multi-drug resistant. With the broth microdilution method, only two isolates (one isolate of B. breve and one isolate of B. longum) were susceptible while 16 isolates were multi-drug resistant. In this study, only two AMR genes were detected: 1) lnu(A) in one isolate of clindamycin-susceptible and lincomycin-resistant Limosilactobacillus reuteri; and 2) tet(W) in one tetracycline-susceptible isolate of B. longum B1-1 and two tetracycline-susceptible isolates and three tetracycline resistant isolates of B. animalis subsp. lactis. Transfer of these two genes via conjugation with a filter mating technique was not observed. These results suggest a need to monitor antimicrobial resistance in newly registered probiotics as well as probiotics with a long history of use.202336746921
539890.9997Characterization and transfer of antimicrobial resistance in lactic acid bacteria from fermented dairy products in China. INTRODUCTION: Lactic acid bacteria (LAB) are commonly found in foods and are also natural intestinal inhabitants in humans and most animals. However, information regarding antimicrobial resistance and the transfer of resistance genes of LAB from fermented dairy products in China is limited. METHODOLOGY: In this study, LAB isolates (n = 82) of Lactobacillus (n = 43) and Streptococcus thermophilus (n = 39) were isolated from 51 commercial fermented food samples in China. All isolates were subjected to pulsed-field gel electrophoresis (PFGE), antimicrobial susceptibility, detecting resistance genes, as well as investigating the transferability of resistance genes. RESULTS: The 43 Lactobacillus isolates yielded 24 PFGE patterns and the 34 isolates of S. thermophilus generated 32 different PFGE patterns. Among the 43 Lactobacillus strains, the most commonly observed resistance was that to streptomycin (83.7%) and gentamycin (83.7%). Among the 39 S. thermophilus strains, the most frequently observed resistance was that to streptomycin (92.3%), gentamycin (87.2%), ciprofloxacin (79.5%), and chloramphenicol (71.8%), whereas the lowest level of resistance was that against erythromycin (7.7%). Antimicrobial resistance genes for erythromycin (emrB), gentamycin (aac(6')-aph(2")), streptomycin (ant(6)), sulfamethoxazole (sulI and sulII), tetracycline (tetM and tetS) were detected in the 18 resistance LAB strains. Conjugation experiments showed that tetM from L. delbrueckii subsp. bulgaricus R6 and tetS from L. plantarum R41 were successfully transferred to L. monocytogenes by filter mating. CONCLUSIONS: LAB strains could potentially act as reservoirs of resistance genes and play an active role in the transfer of resistance to humans via the food chain.201932036349
5538100.9997Phenotypic and genotypic antimicrobial susceptibility pattern of Streptococcus spp. isolated from cases of clinical mastitis in dairy cattle in Poland. Mastitis of dairy cattle is one of the most frequently diagnosed diseases worldwide. The main etiological agents of mastitis are bacteria of the genus Streptococcus spp., in which several antibiotic resistance mechanisms have been identified. However, detailed studies addressing this problem have not been conducted in northeastern Poland. Therefore, the aim of our study was to analyze, on phenotypic and genotypic levels, the antibiotic resistance pattern of Streptococcus spp. isolated from clinical cases of mastitis from dairy cattle in this region of Poland. The research was conducted using 135 strains of Streptococcus (Streptococcus uberis, n = 53; Streptococcus dysgalactiae, n = 41; Streptococcus agalactiae, n = 27; other streptococci, n = 14). The investigation of the antimicrobial susceptibility to 8 active substances applied in therapy in the analyzed region, as well as a selected bacteriocin (nisin), was performed using the minimum inhibitory concentration method. The presence of selected resistance genes (n = 14) was determined via PCR. We also investigated the correlation between the presence of resistance genes and the antimicrobial susceptibility of the examined strains in vitro. The highest observed resistance of Streptococcus spp. was toward gentamicin, kanamycin, and tetracycline, whereas the highest susceptibility occurred toward penicillin, enrofloxacin, and marbofloxacin. Additionally, the tested bacteriocin showed high efficacy. The presence of 13 analyzed resistance genes was observed in the examined strains [gene mef(A) was not detected]. In most strains, at least one resistance gene, mainly responsible for resistance to tetracyclines [tet(M), tet(K), tet(L)], was observed. However, a relationship between the presence of a given resistance gene and antimicrobial susceptibility on the phenotypic level was not always observed.201728601447
5910110.9997Antimicrobial 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
2431120.9997Bacteriocin production, antibiotic susceptibility and prevalence of haemolytic and gelatinase activity in faecal lactic acid bacteria isolated from healthy Ethiopian infants. The objective of this study was to characterise lactic acid bacteria (LAB) isolated from faecal samples of healthy Ethiopian infants, with emphasis on bacteriocin production and antibiotic susceptibility. One hundred fifty LAB were obtained from 28 healthy Ethiopian infants. The isolates belonged to Lactobacillus (81/150), Enterococcus (54/150) and Streptococcus (15/150) genera. Lactobacillus species were more abundant in the breast-fed infants while Enterococcus dominated the mixed-fed population. Bacteriocin-producing LAB species were isolated from eight of the infants. Many different bacteriocins were identified, including one new bacteriocin from Streptococcus salivarius, avicin A (class IIa) from Enterococcus avium, one class IIa bacteriocin from Enterococcus faecalis strains, one unknown bacteriocin from E. faecalis and two unknown bacteriocins from Lactobacillus fermentum strains and the two-peptide gassericin T from Lactobacillus gasseri isolate. Susceptibility tests performed for nine antibiotics suggest that some lactobacilli might have acquired resistance to erythromycin (3 %) and tetracycline (4 %) only. The streptococci were generally antibiotic sensitive except for penicillin, to which they showed intermediate resistance. All enterococci were susceptible to ampicillin while 13 % showed penicillin resistance. Only one E. faecalis isolate was vancomycin-resistant. Tetracycline (51 %) and erythromycin (26 %) resistance was prevalent among the enterococci, but multidrug resistance was confined to E. faecalis (47 %) and Enterococcus faecium (33 %). Screening of enterococcal virulence traits revealed that 2 % were β-haemolytic. The structural genes of cytolysin were detected in 28 % of the isolates in five enterococcal species, the majority being E. faecalis and Enterococcus raffinosus. This study shows that bacteriocin production and antibiotic resistance is a common trait of faecal LAB of Ethiopian infants while virulence factors occur at low levels.201323184155
2903130.9997Soil Bacteria in Urban Community Gardens Have the Potential to Disseminate Antimicrobial Resistance Through Horizontal Gene Transfer. Fifteen soil and 45 vegetable samples from Detroit community gardens were analyzed for potential antimicrobial resistance contamination. Soil bacteria were isolated and tested by antimicrobial susceptibility profiling, horizontal gene transfer, and whole-genome sequencing. High-throughput 16S rRNA sequencing analysis was conducted on collected soil samples to determine the total bacterial composition. Of 226 bacterial isolates recovered, 54 were from soil and 172 from vegetables. A high minimal inhibitory concentration (MIC) was defined as the MIC greater than or equal to the resistance breakpoint of Escherichia coli for Gram-negative bacteria or Staphylococcus aureus for Gram-positive bacteria. The high MIC was observed in 63.4 and 69.8% of Gram-negative isolates from soil and vegetables, respectively, against amoxicillin/clavulanic acid, as well as 97.5 and 82.7% against ampicillin, 97.6 and 90.7% against ceftriaxone, 85.4 and 81.3% against cefoxitin, 65.8 and 70.5% against chloramphenicol, and 80.5 and 59.7% against ciprofloxacin. All Gram-positive bacteria showed a high MIC to gentamicin, kanamycin, and penicillin. Forty of 57 isolates carrying tetM (70.2%) successfully transferred tetracycline resistance to a susceptible recipient via conjugation. Whole-genome sequencing analysis identified a wide array of antimicrobial resistance genes (ARGs), including those encoding AdeIJK, Mex, and SmeDEF efflux pumps, suggesting a high potential of the isolates to become antimicrobial resistant, despite some inconsistency between the gene profile and the resistance phenotype. In conclusion, soil bacteria in urban community gardens can serve as a reservoir of antimicrobial resistance with the potential to transfer to clinically important pathogens, resulting in food safety and public health concerns.202134887843
5428140.9997Antimicrobial resistance and prevalence of resistance genes of obligate anaerobes isolated from periodontal abscesses. BACKGROUND: This study attempts to determine the antimicrobial resistance profiles of obligate anaerobic bacteria that were isolated from a periodontal abscess and to evaluate the prevalence of resistance genes in these bacteria. METHODS: Forty-one periodontal abscess samples were cultivated on selective and non-selective culture media to isolate the oral anaerobes. Their antibiotic susceptibilities to clindamycin, doxycycline, amoxicillin, imipenem, cefradine, cefixime, roxithromycin, and metronidazole were determined using the agar dilution method, and polymerase chain reaction assays were performed to detect the presence of the ermF, tetQ, nim, and cfxA drug resistance genes. RESULTS: A total of 60 different bacterial colonies was isolated and identified. All of the isolates were sensitive to imipenem. Of the strains, 6.7%, 13.3%, 16.7%, and 25% were resistant to doxycycline, metronidazole, cefixime, and amoxicillin, respectively. The resistance rate for both clindamycin and roxithromycin was 31.7%. Approximately 60.7% of the strains had the ermF gene, and 53.3% of the amoxicillin-resistant strains were found to have the cfxA gene. Two nim genes that were found in eight metronidazole-resistant strains were identified as nimB. CONCLUSIONS: In the present study, the Prevotella species are the most frequently isolated obligate anaerobes from periodontal abscesses. The current results show their alarmingly high resistance rate against clindamycin and roxithromycin; thus, the use of these antibiotics is unacceptable for the empirical therapy of periodontal abscesses. A brief prevalence of four resistance genes in the anaerobic bacteria that were isolated was also demonstrated.201423659425
2670150.9997Molecular characterisation and antimicrobial resistance of Streptococcus agalactiae isolates from dairy farms in China. INTRODUCTION: Streptococcus agalactiae (S. agalactiae) is a pathogen causing bovine mastitis that results in considerable economic losses in the livestock sector. To understand the distribution and drug resistance characteristics of S. agalactiae from dairy cow mastitis cases in China, multilocus sequence typing (MLST) was carried out and the serotypes and drug resistance characteristics of the bacteria in the region were analysed. MATERIAL AND METHODS: A total of 21 strains of bovine S. agalactiae were characterised based on MLST, molecular serotyping, antimicrobial susceptibility testing, and the presence of drug resistance genes. RESULTS: The serotypes were mainly Ia and II, accounting for 47.6% and 42.9% of all serotypes, respectively. Five sequence types (STs) were identified through MLST. The ST103 and ST1878 strains were predominant, with rates of 52.4% and 28.6%, respectively. The latter is a novel, previously uncharacterised sequence type. More than 90% of S. agalactiae strains were susceptible to penicillin, oxacillin, cephalothin, ceftiofur, gentamicin, florfenicol and sulfamethoxazole. The bacteria showed high resistance to tetracycline (85.7%), clindamycin (52.1%) and erythromycin (47.6%). Resistant genes were detected by PCR, the result of which showed that 47.6%, 33.3% and 38.1% of isolates carried the tet(M), tet(O) and erm(B) genes, respectively. CONCLUSION: The results of this study indicate that S. agalactiae show a high level of antimicrobial resistance. It is necessary to monitor the pathogens of mastitis to prevent the transmission of these bacteria.202338143824
2909160.9997Determination of the prevalence of antimicrobial resistance genes in canine Clostridium perfringens isolates. Clostridium perfringens is a well documented cause of a mild self-limiting diarrhea and a potentially fatal acute hemorrhagic diarrheal syndrome in the dog. A recent study documented that 21% of canine C. perfringens isolates had MIC's indicative of resistance to tetracycline, an antimicrobial commonly recommended for treatment of C. perfringens-associated diarrhea. The objective of the present study was to further evaluate the antimicrobial susceptibility profiles of these isolates by determining the prevalence of specific resistance genes, their expression, and ability for transference between bacteria. One hundred and twenty-four canine C. perfringens isolates from 124 dogs were evaluated. Minimum inhibitory concentrations of tetracycline, erythromycin, tylosin, and metronidazole were determined using the CLSI Reference Agar Dilution Method. All isolates were screened for three tetracycline resistance genes: tetA(P), tetB(P) and tetM, and two macrolide resistance genes: ermB and ermQ, via PCR using primer sequences previously described. Ninety-six percent (119/124) of the isolates were positive for the tetA(P) gene, and 41% (51/124) were positive for both the tetA(P) and tetB(P) genes. No isolates were positive for the tetB(P) gene alone. Highly susceptible isolates (MIC< or = 4 microg/ml) were significantly more likely to lack the tetB(P) gene. One isolate (0.8%) was positive for the ermB gene, and one isolate was positive for the ermQ gene. The tetM gene was not found in any of the isolates tested. Two out of 15 tested isolates (13%) demonstrated transfer of tetracycline resistance via bacterial conjugation. Tetracycline should be avoided for the treatment of C. perfringens-associated diarrhea in dogs because of the relatively high prevalence of in vitro resistance, and the potential for conjugative transfer of antimicrobial resistance.200616330169
5533170.9997Antibiotic resistance in potential probiotic lactic acid bacteria of fermented foods and human origin from Nigeria. INTRODUCTION: Probiotic lactobacilli are generally recognized as safe (GRAS) and are being used in several food and pharma formulations. However, growing concern of antibiotic resistance in bacterial strains of food origin and its possible transmission via functional foods is increasingly being emphasized. OBJECTIVES: This study screened potential probiotic lactic acid bacteria (LAB) strains for their phenotypic and genotypic antibiotic resistance profiles. METHODS: Susceptibility to different antibiotics was assayed by the Kirby Bauer standard disc diffusion protocol. Both conventional and SYBR-RTq-PCR were used for detection of resistance coding genes. RESULTS: A variable susceptibility pattern was documented against different antibiotic classes. LAB strains irrespective of origin displayed marked phenotypic resistance against cephalosporins, aminoglycosides, quinolones, glycopeptides; and methicillin among beta-lactams with few exceptions. In contrast, high sensitivity was recorded against macrolides, sulphonamides and carbapenems sub-group of beta-lactams with some variations. parC, associated with ciprofloxacin resistance was detected in 76.5% of the strains. Other prevalent resistant determinants observed were aac(6?)Ii (42.1%), ermB, ermC (29.4%), and tetM (20.5%). Six (?17.6%) of the isolates were free from genetic resistance determinants screened in this study. CONCLUSION: Study revealed presence of antibiotic resistance determinants among lactobacilli from both fermented foods and human sources.202337208603
5534180.9997Antibiotic 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
2671190.9997Toxinotyping and molecular characterization of antimicrobial resistance in Clostridium perfringens isolated from different sources of livestock and poultry. The present study was designed to understand the presence of antimicrobial resistance among the prevalent toxinotypes of Clostridium perfringens recovered from different animals of Tamil Nadu, India. A total of 75 (10.76%) C. perfringens were isolated from 697 multi-species fecal and intestinal content samples. C. perfringens type A (90.67%), type C (2.67%), type D (4%) and type F (2.67%) were recovered. Maximum number of isolates were recovered from dog (n = 20, 24.10%) followed by chicken (n = 19, 5.88%). Recovered isolates were resistant to gentamicin (44.00%), erythromycin (40.00%), bacitracin (40.00%), and tetracycline (26.67%), phenotypically and most of the isolates were found to be resistant to multiple antimicrobials. Genotypic characterization revealed that tetracycline (41.33%), erythromycin (34.66%) and bacitracin (17.33%) resistant genes were present individually or in combination among the isolates. Combined results of phenotypic and genotypic characterization showed the highest percentage of erythromycin resistance (26.66%) among the isolates. None of the isolates showed amplification for lincomycin resistance genes. The correlation matrix analysis of genotypic resistance showed a weak positive relationship between the tetracycline and bacitracin resistance while a weak negative relationship between the tetracycline and erythromycin resistance. The present study thus reports the presence of multiple-resistance genes among C. perfringens isolates that may be involved in the dissemination of resistance to other bacteria present across species. Further insights into the genome can help to understand the mechanism involved in gene transfer so that measures can be taken to prevent the AMR spread.202133220406