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Eighty enterococcal isolates (E. faecium, n=38, E. faecalis, n=42) from diseased farm animals (swine, cattle, poultry) in Lithuania have been studied for the prevalence of antibiotic resistance and for resistance and virulence genetic determinants. 86% of E. faecium and 71% of E. faecalis isolates were multidrug resistant (resistant to three or more unrelated antibiotics). Resistance to aminoglycosides, tetracycline and erythromycin was found most frequently in both species (61%, 69%) and was linked to aph(3’)-IIIa, aac(6’)-Ie-aph(2”)-Ia, ant(6)-Ia (aminoglycoside resistance), tetM, tetL (tetracycline resistance), ermA, ermB (erythromycin resistance) gene combinations, which were supplemented with chloramphenicol resistance genes catA7, catA8 (E. faecalis) and catA9 (E. faecium). All E. faecalis isolates harboured genes coding for virulence factors agg, esp, fsr gelE alone or in combinations with the high prevalence of esp gene in isolates from cattle (63%) and pigs (79%). The origin- dependent incidence of agg gene variants prgB and asp1 was observed. The results indicate the existence of a large pool of potentially virulent and multidrug resistant E. faecalis in diseased farm animals posing risk to humans.
Antimicrobial susceptibility of isolated enterotoxigenic E. coli, Salmonella enterica serovar Choleraesuis, Pasteurella multocida and Streptococcus suis from pigs was tested in the Veterinary Institute of the Lithuanian Veterinary Academy. Commensal E. coli and Enterococcus faecalis were also included in the testing as commensal bacteria. Clinical and pathological material was investigated from various regions of the country. Isolation and identification of bacteria was done using common methods. The agar diffusion method according to NCCLS guidelines was applied for antimicrobial susceptibility testing. Enterotoxigenic E. coli showed the highest resistance to tetracycline (67%), ampicillin (52%) and sulfamethoxazole-trimethoprim (43%). Not less than 90% of these bacteria were susceptible to colistin, florfenicol and ceftiofur. Salmonella Choleraesuis demonstrated the highest resistance to tetracycline (53%). Florfenicol, ceftiofur and enrofloxacin were effective against most strains of salmonella. Pasteurella multocida in most cases were susceptible to all the tested antimicrobials, however 20% of the isolates were resistant to sulfamethoxazole-trimethoprim. Streptococcus suis demonstrated the highest resistance to tetracycline (43%), lincomycin (40%), sulfamethoxazole- -trimethoprim (40%), and erythromycin (30%). Ceftiofur was the most effective against S. suis. Commensal E. coli showed less resistance than enterotoxigenic E. coli, however not less than 25% of isolates were resistant to tetracycline, sulfamethoxazole-trimethoprim and ampicillin. All the tested Enterococcus faecalis were susceptible to vancomycin and ceftiofur and 80% of enterococci were resistant to tetracycline.
During 2011, 160 nasal samples were taken from pigs on 8 different farms in Lithuania. Four methicillin-resistant Staphylococcus aureus (MRSA) isolates were obtained. The isolates were ST398, spa type t011 and SCCmec V and none carried the lukF/lukS genes. Strains were resistant to tetracycline, attributed to tetK and tetM genes, and to erythromycin owing to the ermB gene. One MRSA strain was resistant to trimethoprim/sulfamethoxazole and carried the dfrK gene. This is the first report on the presence and characteristics of livestock-associated MRSA isolated from pigs in Lithuania.
The objectives of this study were to determine the prevalence and antimicrobial resistance of coagulase-negative staphylococci (CNS) isolated from dairy cows with subclinical mastitis. Antimicrobial resistance in staphylococci were evaluated by breakpoint values specific to the species (EU-CAST). The presence of resistance-encoding genes was detected by multiplex PCR. A total of 191 CNS isolates were obtained. The CNS isolates were typically resistant to penicillin (67.4%), tetracyc-line (18.9%), and erythromycin (13.7%). CNS isolates (78.0%) were resistant to at least one antimicrobial compound, and 22.0% were multiresistant. The multiresistant isolates were predominantly Staphylococcus chromogenes (28.6%), Staphylococcus warneri (19%) and Staphylococcus haemolyticus (14.3%). According to MIC pattern data, multiresistant isolates showed the highest resistance (p<0.05) rates to penicillin (85.7%), tetracycline (66.7%), and erythromycin (48.2%), but all of them were sensitive to daptomycin, oxacillin, qiunupristin/dalfopristin, and vancomycin. S. chromogenes (9.5%), S. haemolyticus (4.8%), and S. capitis ss capitis (2.4%) strains were resistant to methicillin; their resistance to oxacillin and penicillin was more than 8 mg/l. A high rate of resistance to penicillin was linked to a blaZ gene found in 66.6% of the isolated multiresistant CNS strains. Resistance to tetracycline via the tetK (38.1%) gene and penicillin via the mecA (23.8%) gene were detected less frequently. Gene msrAB was responsible for macrolides and lincosamides resistance and detected in 28.6% of the CNS isolates. Antimicrobial resistance genes were identified more frequently in S. epidermidis, S. chromogenes, and S. warneri.
The aim of the study was to isolate cultivable gut microbiota from European pond turtles kept at the Lithuanian Zoo and to determine antimicrobial resistance of the isolates. The study subjects included 8 elderly turtles living at the Lithuanian Zoo for about 50 years as well as their offspring – 24 young individuals (1-2 years old) that were hatched at the same zoo. Animals were not exposed by treatment with antimicrobials during the last 3 years. Gut samples were taken from the cloaca and inoculated onto universal media. Isolates then were identified using sequence analysis of 16S rRNA. The antimicrobial susceptibility testing was performed using the agar diffusion method according to Kirby-Bauer. Clinical breakpoints according to CLSI whenever possible, were used for interpretation of susceptibility. Bacterial isolates resistant to at least three antimicrobials of different classes were treated as multi-resistant. Fifty-two bacterial isolates were obtained and identified from turtle gut samples. The most prevalent genera included Aeromonas, Chryseobacterium and Citrobacter. Fifty percent of the isolates obtained from elderly turtles (CI 95% – 19.01-80.99) and 54.8% (CI 95% – 39.75-69.85) of the isolates from young animals were identified as multi-resistant. The most common resistance rates of the isolates from both groups of the turtles were observed toward ampicillin (86.6%), ciprofloxacin (61.5%) and gentamicin (40.4%). The lowest number of resistant isolates were detected toward combination of sulfamethoxazole-trimethoprim (26.9%). The study revealed that European pond turtles kept in captivity are carriers of multi-resistant bacteria however, further studies need to be performed to investigate whether the resistant microorganisms are natural microbiota for this species or they were acquired in the zoo.
The aim of this study was to characterize Staphylococcus pseudintermedius for its antimicrobial resistance and virulence factors with a special focus on methicillin-resistant (MRSP) strains isolated from sick dogs in Lithuania. Clinically sick adult dogs suffering from infections (n=214) and bitches with reproductive disorders (n=36) from kennels were selected for the study. Samples (n=192) from the 250 tested (76.8%) dogs were positive for Staphylococcus spp. Molecular profiling using the species-specific nuc gene identified 51 isolates as S. pseudintermedius (26.6% from a total number of isolated staphylococci) of which 15 isolates were identified as MRSP. Ten MRSP isolates were isolated from bitches with reproductive disorders from two large breeding kennels. Data on susceptibility of S. pseudintermedius to different antimicrobials revealed that all isolates were susceptible to vancomycin, daptomycin and linezolid. Two isolates (3.9%) were resistant to rifampicin. A high resistance was seen towards penicillin G (94.1%), tetracycline (64.7%) and macrolides (68.7%). Resistance to fluoroquinolones ranged from 25.5% (gatifloxacin) to 31.4% (ciprofloxacin). The most prevalent genes encoding resistance included blaZ, aac(6’)-Ie-aph(2’’)-Ia, mecA, and tet(M). The Luk-I gene encoding a leukotoxin was detected in 29% of the isolates, whereas the siet gene encoding exfoliative toxin was detected in 69% of the S. pseudintermedius isolates. This report of MRSP in companion animals represents a major challenge for veterinarians in terms of antibiotic therapy and is a concern for both animal and public health.
The aim of the study was to isolate gram-positive cocci from cows with mastitis and to determine their resistance to beta-lactamic antibiotics. Eight hundred and nine strains were isolated and identified as staphylococci (n=516), streptococci (n=199) and enterococci (n=94) from sub-clinical and clinical cases of bovine mastitis in Lithuania. The most common causative agents of udder disease included: S. epidermidis (n=176), S. aureus (n=176), S. agalactiae (n=134), S. hyicus (136) and E. hirae (n=68). Isolates were analysed for antimicrobial resistance to penicillin, ampicillin, amoxicillin, cephalothin, cephalexin, amoxicillin + clavulanate. The susceptibility patterns were analysed using the agar disk diffusion method. S. aureus showed the highest level of resistance to amoxicillin (81.3%), penicillin (76.7%) and ampicillin (78.4%). The corresponding values for CNS strains were 59.7%, 59.7% and 50.6% against penicillin, ampicillin and amoxicillin respectively. Streptococci were the most frequently resistant to amoxicillin (29.3%), and enterococci to penicillin (27%), amoxicillin (27.5%) and amoxicillin/clavulanic acid (23.8%). The resistance of all tested mastitis pathogens to aminopenicillins and penicillin highly correlated (r=0.83). Compared with other antibiotics, amoxicillin and clavulanic acid combination tended to be more effective (p<0.05) against all tested bacteria in vitro. However, S. aureus, in 38.1% of cases, was resistant to this combination of antimicrobials. This study demonstrates that S. epidermidis, S. aureus, S. hyicus, S. agalactiae and E. hirae remain the most frequent mastitis causative agents on Lithuanian cattle farms. The highest resistance in vitro to penicillins was demonstrated by S. aureus, S. hyicus and S. intermedius. Resistance to cephalosporins remains low, irrespective of bacterial species of gram-positive cocci.
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