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Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp. isolated from dairy and pharmaceutical products.

The outlines of antibiotic resistance of some probiotic microorganisms were studied. This study was conducted with the double purpose of verifying their ability to survive if they are taken simultaneously with an antibiotic therapy and to increase the selective properties of suitable media for the isolation of samples containing mixed bacterial populations. We isolated from commercial dairy and pharmaceutical products, 34 strains declared as probiotics, belonging to the genera Bifidobacterium and Lactobacillus, and 21 strains of starter culture bacteria. All the microorganisms have been compared by electrophoresis of the soluble proteins for the purpose of identifying them. A Multiplex-PCR with genus- and species-specific primers was used to detect for Bifidobacterium animalis subsp. lactis presence. All bifidobacteria were B. animalis subsp. lactis except one Bifidobacterium longum. Sometimes the identification showed that the used strain was not the one indicated on the label. The lactobacilli were Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus delbrueckii subsp. bulgaricus. The streptococci were all Streptococcus thermophilus. The minimal inhibitory concentration (MIC) of 24 common antibiotic substances has been valued by the broth microdilution method. All tested strains were susceptible to ampicillin, bacitracin, clindamycin, dicloxacillin, erytromycin, novobiocin, penicillin G, rifampicin (MIC(90) ranging from 0.01 to 4 microg/ml); resistant to aztreonam, cycloserin, kanamycin, nalidixic acid, polymyxin B and spectinomycin (MIC(90) ranging from 64 to >1000 microg/ml). The susceptibility to cephalothin, chloramphenicol, gentamicin, lincomycin, metronidazole, neomycin, paromomycin, streptomycin, tetracycline and vancomycin was variable and depending on the species.

Animals↗

Identification of the pharmaceuticals for human use contaminating the Italian aquatic environment.

A predictive approach seems useful to identify pharmaceuticals in the environment and give an idea of overall levels of contamination, so as to restrict monitoring to those molecules most likely to be contaminants. We propose an approach based on two parts. The first is to rank the molecules according to the predicted environmental loads; the second is to refine the list by analysing the pharmaceuticals in sewage treatment plants (STPs) and comparing the concentrations with levels previously measured in surface water. This approach identified a restricted group of priority pollutants (ofloxacin, furosemide, atenolol, hydrochlorothiazide, carbamazepine, ibuprofen, spiramycin, bezafibrate, erythromycin, lincomycin and clarithromycin) in the aquatic environment in Italy, for further studies and monitoring.

Adrenergic beta-Antagonists↗

Pulse-field gel electrophoresis typing of methicillin-resistant Staphylococcus aureus strains susceptible to aminoglycosides isolated from 1993 to 2002.

Methicillin-resistant Staphylococcus aureus (MRSA) susceptible strains to aminoglycosides (AS-MRSA) have been increasingly isolated in the Albert Cheneiver Hospital during the past 10 years. The aim of this study was first, to analyse the genotypes and the profiles of resistance to antibiotics and second to compare the AS-MRSA with the MRSA resistant to gentamicin (GR-MRSA) and with MRSA resistant to kanamycin and tobramycin, but susceptible to gentamicin (GS-MRSA), previously studied in our laboratory. All the AS-MRSA consecutively isolated from clinical samples (carriage isolates excluded) from 01/01/1993 to 31/12/2002 (33 isolates) were typed by DNA macrorestriction. Their susceptibilities to other anti-staphylococcal drugs (erythromycin, lincomycin, tetracycline, rifampicin, fusidic acid and fosfomycin) were studied by the French standard disk method. The 33 strains showed a heterogeneous resistance to oxacillin and fell into five phenotypes. The main phenotype (51.5% of the AS-MRSA strains) was susceptible to the six antibiotics studied. DNA macrorestriction defined 24 genotypes (percentage similarity <80%). Among them 16 genotypes contained only one strain each, and none contained more than three isolates. Conversely the comparison with GR-MRSA and GS-MRSA isolated during the same period showed that the strains were not closely linked. The diversity of our isolates showed that it was not an epidemic phenomenon, in contrast to the results of similar studies. Our findings may be explained by the patients coming mostly from different hospital units. This work indicates the need for further studies on the genome, to determine whether AS-MRSA have derived from strains that occurred before aminoglycosides came into clinical use.

Aminoglycosides↗

Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

Animals↗

Acquired antimicrobial resistance in the intestinal microbiota of diverse cat populations.

The aim of this study was to investigate the prevalence of acquired antimicrobial resistance in the resident intestinal microbiota of cats and to identify significant differences between various cat populations. Escherichia coli, Enterococcus faecalis, E. faecium and Streptococcus canis were isolated as faecal indicator bacteria from rectal swabs of 47 individually owned cats, 47 cattery cats and 18 hospitalised cats, and submitted through antimicrobial sensitivity tests. The results revealed that bacteria isolated from hospitalised and/or cattery cats were more frequently resistant than those from individually owned cats. E. coli isolates from hospitalised cats were particularly resistant to ampicillin, tetracycline and sulfonamide. Both enterococci and streptococci showed high resistance to tetracycline and in somewhat lesser extent to erythromycin and tylosin. Most E. faecium isolates were resistant to lincomycin and penicillin. One E. faecalis as well as one E. faecium isolate from hospitalised cats showed 'high-level resistance' (MIC > 500 microg/ml) against gentamicin, a commonly used antimicrobial agent in case of human enterococcal infections. The results of this research demonstrate that the extent of acquired antimicrobial resistance in the intestinal microbiota of cats depends on the social environment of the investigated population. It is obvious that the flora of healthy cats may act as a reservoir of resistance genes.

Animals↗

External quality assurance system for antibiotic resistance in bacteria of animal origin in Europe (ARBAO-II), 2003.

Initiated in 2003 by the European Union, ARBAO-II aims to establish a monitoring of antimicrobial susceptibility among the veterinary laboratories in all European countries based on validated methodologies. This includes an external quality control system for the most important bacterial pathogens. In 2003 two trials were performed. The first on Salmonella and Escherichia coli involved 31 laboratories in 18 countries and the second on Staphylococcus and Streptococcus included 22 laboratories in 15 countries. For the E. coli strains, 92.8% of the results complied with the reference MICs, for Salmonella 93.7%, for Streptococcus 80.4% and for Staphylococcus 93.1%. Most problems were observed when testing florfenicol (79.2% correct), gentamicin (84.2%) and amoxicillin + cl (84.9%) in E. coli; streptomycin in Salmonella (62.5%); gentamicin (56.7%), lincomycin (71.4%), clindamycin (75.4%), TMP + sulfonamides (75.7%) and chloramphenicol (78.5%) in Streptococcus; erythromycin (81.5%) and oxacillin (78.2.5%) in Staphylococcus. A few laboratories caused most deviations. However, there was no correlation between good performance for one bacterial group and good performance for other groups. This study showed that most laboratories are capable of performing correct susceptibility testing for E. coli and Salmonella, even though performance of some laboratories can be improved, and that some problems exist for Staphylococcus and Streptococcus. This clearly shows the need for continuous harmonisation of methodologies within the EU.

Animals↗

Disinfection of swine wastewater using chlorine, ultraviolet light and ozone.

Veterinary antibiotics are widely used at concentrated animal feeding operations (CAFOs) to prevent disease and promote growth of livestock. However, the majority of antibiotics are excreted from animals in urine, feces, and manure. Consequently, the lagoons used to store these wastes can act as reservoirs of antibiotics and antibiotic-resistant bacteria. There is currently no regulation or control of these systems to prevent the spread of these bacteria and their genes for antibiotic resistance into other environments. This study was conducted to determine the disinfection potential of chlorine, ultraviolet light and ozone against swine lagoon bacteria. Results indicate that a chlorine dose of 30 mg/L could achieve a 2.2-3.4 log bacteria reduction in lagoon samples. However, increasing the dose of chlorine did not significantly enhance the disinfection activity due to the presence of chlorine-resistant bacteria. The chlorine resistant bacteria were identified to be closely related to Bacillus subtilis and Bacillus licheniformis. A significant percentage of lagoon bacteria were not susceptible to the four selected antibiotics: chlortetracycline, lincomycin, sulfamethazine and tetracycline (TET). However, the presence of both chlorine and TET could inactivate all bacteria in one lagoon sample. The disinfection potential of UV irradiation and ozone was also examined. Ultraviolet light was an effective bacterial disinfectant, but was unlikely to be economically viable due to its high energy requirements. At an ozone dose of 100 mg/L, the bacteria inactivation efficiency could reach 3.3-3.9 log.

Animal Husbandry↗

Epidemiosurveillance of antimicrobial compound resistance of Staphylococcus intermedium clinical isolates from canine pyodermas.

In a retrospective study, 131 Staphylococcus intermedius strains isolated from apparently healthy dogs, and 187 Staphylococcus intermedius strains isolated from dog pyodermas in the clinical microbiology laboratory at the National Veterinary School in Nantes, during three successive periods: 1986-87, 1992-93 and 1995-96, were investigated and compared for their antimicrobial susceptibility. Results indicated that 60% to 65% of the strains were susceptible to Chloramphenicol and Doxycyclin, 65% to 80% of the strains were susceptible to macrolides (Erythromycin, Lincomycin and Clindamycin) and to Trimethoprim/Sulfonamide association. More than 95% of the strains were susceptible to three betalactamins tested: Oxacillin, Amoxycillin/Clavulanic acid, Cephalexin, to Gentamicin, to Fucidic Acid and to two quinolones: Enrofloxacin and Marbofloxacin. This last group is made up of choice antibacterials for the treatment of dog pyoderma. Many different resistance patterns were observed in each period with no really predominant profile, because of low plasmidic vs chromosomal balance of the genetic basis of antibacterial resistance in Staphylococcus intermedius. However, the proportion of multiresistant (> or = 3 drugs) strains increased from 10.8% in the first period, to 28% in the third period. This increased frequency of resistance suggests strongly that, as in Staphylococcus aureus human infections, the prescription of antibiotic compounds increases the prevalence of resistant strains.

Aminoglycosides↗

Associations between off-label feed additives and farm size, veterinary consultant use, and animal age.

Data from the United States National Swine Survey collected by the National Animal Health Monitoring System were used to describe the use of feed additives in swine feeds. Data were collected from 710 farms. The concentration of feed additives expressed in grams per ton of complete feed was described by stage of production, and the use of feed additives above the labeled treatment levels (i.e. off-label) was identified. Of the 3328 feeds, about 79% contained feed additives used in the labeled manner. For all classes of pigs, the prevalence of labeled feed additive use was greater than 75%. Penicillin was used according to its label most often, followed by apramycin, bacitracin, tetracyclines, lincomycin, and tylosin. Carbadox had the highest prevalence of off-label use. Of the 699 feeds that included feed additives in an off-label manner, about 57% included additives at greater than the recommended concentrations or were fed to an incorrect class of pig. About 56% of the feeds had off-label combinations of additives. Small farms were more likely to use rations with no feed additives than intermediate or large farms (P < 0.001). Of those farms using feed additives, the odds of a small farm using all feed additives in the labeled manner was 7.7 times that of an intermediate or large farm (P < 0.0001). After controlling for herd size, producers who used a veterinary consultant were 2.1 times more likely to use feeds with feed additives (P < 0.0001).

Aging↗

Susceptibility of Listeria monocytogenes isolated from food in Italy to antibiotics.

The susceptibility of 148 strains of Listeria monocytogenes isolated from food to antibiotics currently used in veterinary and human therapy was determined by standard agar dilution and disk diffusion methods. The antibiotics included amikacin, amoxicillin, cefazolin, chloramphenicol, erythromycin, flumequine, fosfomycin, gentamicin, kanamycin, lincomycin, oxytetracycline, rifampicin, spiramycin, streptomycin, tetracycline, tobramycin and vancomycin. Soussy's breakpoints and MIC(50)-MIC(90) values were used to classify the strains into sensitive, moderately sensitive and resistant groups. This work is part of a wider surveillance program on listeriosis started in Italy in 1995.

Anti-Bacterial Agents↗

Molecular structure and evolution of the conjugative multiresistance plasmid pRE25 of Enterococcus faecalis isolated from a raw-fermented sausage.

Plasmid pRE25 from Enterococcus faecalis transfers resistances against kanamycin, neomycin, streptomycin, clindamycin, lincomycin, azithromycin, clarithromycin, erythromycin, roxithromycin, tylosin, chloramphenicol, and nourseothricin sulfate by conjugation in vitro to E. faecalis JH2-2, Lactococcus lactis Bu2, and Listeria innocua L19. Its nucleotide sequence of 50237 base pairs represents the largest, fully sequenced conjugative multiresistance plasmid of enterococci (Plasmid 46 (2001) 170). The gene for chloramphenicol resistance (cat) was identified as an acetyltransferase identical to the one of plasmid pIP501 of Streptococcus agalactiae. Erythromycin resistance is due to a 23S ribosomal RNA methyl transferase, again as found in pIP501 (ermB). The aminoglycoside resistance genes are packed in tandem as in transposon Tn5405 of Staphylococcus aureus: an aminoglycoside 6-adenyltransferase, a streptothricin acetyl transferase, and an aminoglycoside phosphotransferase.). Identical resistance genes are known from pathogens like Streptococcus pyogenes, S. agalactiae, S. aureus, Campylobacter coli, Clostridium perfringens, and Clostridium difficile. pRE25 is composed of a 30.5-kbp segment almost identical to pIP501. Of the 15 genes involved in conjugative transfer, 10 codes for putative transmembrane proteins (e.g. trsB, traC, trsF, trsJ, and trsL). The enterococcal part is joined into the pIP501 part by insertion elements IS1216V of E. faecium Tn1545 (three copies), and homologs of IS1062 (E. faecalis) and IS1485 (E. faecium). pRE25 demonstrates that enterococci from fermented food do participate in the molecular communication between Gram-positive and Gram-negative bacteria of the human and animal microflora.

Animals↗

Molecular characterization of the prokaryotic efp gene product involved in a peptidyltransferase reaction.

The translation factor EF-P is required for efficient prokaryotic peptide bond synthesis on 70S ribosomes from fMet-tRNAfMet. This protein has been purified from Escherichia coli cells and the gene, efp, encoding it has been cloned and sequenced. We have isolated recombinant clones which overexpress a protein that co-migrates with purified EF-P upon SDS-PAGE analysis. Using these clones, we report the purification, crystallization and initial characterization of the efp gene product. The mechanism by which EF-P stimulates peptide-bond synthesis was studied using several antibiotics that inhibit translocation, peptide-bond synthesis and decoding. The stimulation of peptidyltransferase by EF-P was not inhibited by antibiotics that affect translocation and occupation of the A site (in the elongation state), ie thiostrepton, viomycin, neomycin and fusidic acid but was inhibited by streptomycin as well as by inhibitors of peptidyltransferase, chloramphenicol and lincomycin. This observation and the requirement for L16 but not for the L7/L12 nor L6 or L11 r-proteins suggest that the binding site for EF-P may overlap the peptidyltransferase center of the ribosome.

Anti-Bacterial Agents↗

Characterization of Staphylococcus aureus isolates from oral surgical outpatients compared to isolates from hospitalized and non-hospitalized individuals.

In 160 patients, 114 strains of Staphylococcus aureus were isolated from gingival mucosa, saliva, throat and nose. The patients were divided into 4 groups: one group from an oral surgery clinic for outpatients, one group from a dental clinic, one group from a general surgery clinic for inpatients and one group from a clinic for chronically ill and aged patients. The highest frequency of staphylococcal carriers was found in the outpatient groups (oral surgery and dental clinic), 55% and 45%, respectively. Antibiotic sensitivity testing revealed the majority of the strains to be penicillin resistant, but sensitive to isoxazolyl-penicillins, clindamycin and lincomycin. 50% of the strains produced penicillinase. About 90% of all strains produced lipase, nuclease and a haemolysin most active on rabbit erythrocytes. Human and sheep erythrocytes were lysed by 70% and 46% of the strains, respectively. 77% of the strains were bacteriolytic active. No strain produced lecitinase or elastase. No significant difference was found between the 4 groups in the formation of any of these factors. Phage-type I and III dominated but there was no correlation to enzyme production or patient groups. Thus the 4 patient groups were colonized with strains of Staphylococcus aureus that showed mainly the same pathogenetic factors.

Adolescent↗

Common childhood bacterial infections.

Acute bacterial infections are a common cause of pediatric visits to the emergency department. The diagnosis and treatment of pharyngitis, otitis media, pneumonia, urinary tract infection, acute bacterial lymphadenitis and gastroenteritis are reviewed. The authors propose the use of a limited number of antibiotic agents including penicillin, ampicillin, erythromycin, lincomycin, cephalexin, a sulfonamide and tetracycline to improve efficiency and quality of care and to allow physicians to become familar with the drugs' characteristics, indications, dosage and side effects.

Anti-Bacterial Agents↗

[Antibiotic resistance of Staphylococcus aureus in urban experience: 6 month study in Aquitaine].

Antibiotic resistance of Staphylococcus aureus has been surveyed by eight city laboratories of the Aquitaine area, during a six month-period (january to june 1998). Antibiotic susceptibility testing has been performed by the disk diffusion method, and the results have been collected and analysed using the SIRscan system. After elimination of the redundant strains, a total of 747 isolates has been retained. They were mainly isolated from pus (64%) collected from patients of the community (40%) or hospitalized in 30 private clinics or nursing homes. The percentages of resistant strains (community/institutions) were: benzylpenicillin: 90% (87/92%), oxacillin: 39% (23/50%), kanamycin: 37% (22/47%); gentamicin: 13% (8/16%), tobramycin: 37% (21/47%), amikacin: 21% (13/27%); netilmicin: 6% (5/7%), erythromycin: 33% (30/35%), spiramycin: 72% (77/69%), lincomycin: 24% (17/29%), pristinamycin: 2% (1/2%), tetracycline: 17% (14/19%); pefloxacin: 40% (25/50%), fosfomycin: 9% (6/12%), rifampicin: 10% (7/13%), fusidic acid: 14% (11/15%), cotrimoxazole and vancomycin: 0%. Meticillin-susceptible strains of S. aureus were mostly sensitive to other antibiotics (< or = 6% resistant strains, except for erythromycin: 22%). Among meticillin-resistant S. aureus, heterogeneous strains with a KT phenotype, and various resistance patterns to the remaining antibiotics were predominant (61%), compared to the homogeneous strains with a KTG phenotype and multiresistant to the other antibiotics (32%). The frequencies of resistant strains were highly variable depending on the specimen, the laboratory and the health care institution, except for cotrimoxazole and vancomycin which were always active.

Anti-Bacterial Agents↗

Antimicrobial susceptibility of Mycoplasma hyorhinis.

A broth microdilution technique was used to determine the antimicrobial susceptibility of 15 field isolates of Mycoplasma hyorhinis to 10 antimicrobial agents, representative of different classes, and contrasting newer agents to existing ones. For the macrolides, the MIC(90) for tylosin and tilmicosin was 1 and 4 microg/ml, respectively, but was > or = 16 microg/ml for erythromycin. Tetracycline, lincomycin and enrofloxacin each had an MIC(90) of 2 microg/ml. The mycoplasma had similar levels of susceptibility to the aminoglycoside and aminocyclictol classes exhibiting an MIC(90) of 4 microg/ml for gentamicin and 2 microg/ml for spectinomycin. The isolates exhibited high MICs to trimethoprim/sulfamethoxazole with an MIC(90) > or = 16/304 microg/ml. In summary, M. hyorhinis isolates from the US had low MICs against a variety of antimicrobials tested, with the exception of erythromycin and trimethoprim/sulfamethoxazole.

Animals↗

Characterization of four Flavobacterium columnare (Flexibacter columnaris) strains isolated from tropical fish.

Four Flavobacterium columnare strains (AJS 1-4) were isolated from black mollies (Poecilia sphenops) and platies (Xiphophorus maculatus), showing white spots on the back, head and skin ulcers. The isolates developed characteristic rhizoid yellow pigmented colonies on Shieh agar and typical growth in Shieh broth. They were Gram-negative, filamentous bacteria exhibiting flexing movements. When compared to F. columnare strains isolated from temperate fish, it was noted that the four strains originating from tropical aquarium fish are more capable of growing at higher temperatures, the opposite being true for the strains isolated from temperate fish. Biochemical characterization and agglutination tests proved that the isolated strains could be classified as F. columnare. Low minimal inhibitory concentration (MIC) values were found for chloramphenicol, erythromycin, furazolidone, kanamycin, lincomycin, nalidixic acid, oxytetracycline and streptomycin. MIC values were high for colistin, sulfamethoxazole and neomycin. Pathogenicity studies were performed on black mollies. When these animals were submersed in an infective solution of the F. columnare strains, a marked difference in virulence was noted among the four isolated strains, strain AJS 1 being the most virulent one and strain AJS 4 being of low virulence.

Agglutination Tests↗