Effect of supplementation with Enterococcus faecium (SF68) on immune functions in cats

T evaluate the effect of supplementation with Enterococcus faecium strain SF68 (NCIMB10415) on immune function, responses to a multivalent vaccine were investigated in kittens given palatability enhancer with or without E. faecium SF68 daily. E. faecium SF68 was detected in the feces of seven of nine treated cats. Supplementation of kittens with E. faecium SF68 did not affect developmental parameters. The percentage of CD4+ lymphocytes was significantly higher in the treatment group. There were no statistical differences in measurements of any other nonspecific or specific immune parameters between groups.     

The role of Acinetobacter baumannii as a nosocomial pathogen for dogs and cats in an intensive care unit

Acinetobacter baumannii is a nosocomial pathogen associated with high morbidity and mortality in humans. Whereas infections with strains of Acinetobacter species have been reported in various situations, the importance of A baumannii as a nosocomial pathogen in veterinary hospitals has not been studied so far. In this retrospective case series, we describe 17 dogs and 2 cats from which A baumannii had been isolated during a 21/2-year period. In 7 dogs, A baumannii induced systemic signs of illness, whereas 12 animals showed signs of local infection. In all animals with systemic infection, and in 2 with localized infection, A baumannii contributed to the death of the animal or contributed to euthanasia; the remaining 8 dogs and both cats recovered. Molecular typing of the isolates with restriction polymorphisms of ribosomal DNA provided evidence of nosocomial spread of this pathogen and for the presence of several strains of A baumannii in the hospital environment.     

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.     

First report of multiresistant, mecA-positive Staphylococcus intermedius in Europe: 12 cases from a veterinary dermatology referral clinic in Germany

Resistance to cephalosporins and/or fluoroquinolones by Staphylococcus intermedius has remained low in Europe, with effective drugs generally available for systemic therapy in pets. However, multiresistant, mecA-positive S. intermedius isolated from dogs and cats is now emerging in Europe. Twelve S. intermedius isolates, highly resistant to at least five antimicrobial classes, were isolated from skin and ear infections in 11 dogs and a cat. The 12 isolates represented 23% of all S. intermedius submissions from one veterinary dermatology referral clinic in northern Germany to veterinary diagnostic laboratories during an 18-month period and resistance included cefalexin, methicillin and enrofloxacin. The animals had been referred to the clinic with recurrent superficial pyoderma, deep pyoderma, pododermatitis or chronic otitis, all unresponsive to systemic beta-lactam-antibiotics or fluoroquinolones. Infection resolved in 10 dogs and the cat on a combination of antimicrobial treatment and correction of underlying causes. Four dogs and a cat required systemic and topical therapy; in six dogs topical antimicrobial therapy alone was successful. Phenotypic and genotypic characteristics of the S. intermedius isolates were determined; species identification was confirmed by polymerase chain detection of thermonuclease genes (nuc) and the presence and expression of the gene conferring resistance to all beta-lactam antibiotics (mecA) were demonstrated in all; based on pulsed-field gel electrophoresis, six were indistinguishable, the others closely or possibly related. The emergence of multiresistant, mecA-positive S. intermedius in Europe is alarming. Zoonotic implications, awareness among veterinary laboratories and strategies for the use of antimicrobials in small animal practice need to be considered.     

Antimicrobial drug use and resistance in dogs

Fifteen years (1984-1998) of records from a Veterinary Teaching Hospital were analyzed to determine whether antimicrobial drug resistance in coagulase-positive Staphylococcus spp. (S. aureus, S. intermedius) isolated from clinical infections in dogs has increased, and whether there has been a change in the species of bacteria isolated from urinary tract infections in dogs. In coagulase-positive Staphylococcus spp., a complex pattern showing both increases and decreases of resistance to different classes of antimicrobial drugs was observed, reflecting the changing use of different antimicrobial drug classes in the hospital over a similar period (1990-1999). In canine urinary tract infections identified from 1984 to 1998, an increase in the incidence of multiresistant Enterococcus spp. was apparent, with marginal increases also in incidence in Enterobacter spp. and in Pseudomonas aeruginosa, both of which, like Enterococcus spp., are innately antimicrobial-resistant bacteria. A survey of directors of veterinary teaching hospitals in Canada and the United States identified only 3 hospitals that had any policy on use of "last resort" antimicrobial drugs (amikacin, imipenem, vancomycin). Evidence is briefly reviewed that owners may be at risk when dogs are treated with antimicrobial drugs, as well as evidence that some resistant bacteria may be acquired by dogs as a result of antimicrobial drug use in agriculture. Based in part on gaps in our knowledge, recommendations are made on prudent use of antimicrobial drugs in companion animals, as well as on the need to develop science-based infection control programs in veterinary hospitals.     

Antimicrobial and heavy metal resistance in commensal enterococci isolated from pigs

Antibiotic resistance in animal isolates of enterococci is of public health concern because of the risk of transfer of antibiotic resistance isolates or resistance determinants to consumers via the food chain. In this study, phenotypic and genotypic resistance in 192 pig isolates of enterococci to ampicillin, avilamycin, avoparcin, bacitracin, flavophospholipol, gentamicin, narasin, tetracycline, tiamulin, tylosin, vancomycin, virginiamycin, copper and zinc were investigated by susceptibility test and molecular methods. Resistance rates varied between the species but all isolates were susceptible to ampicillin, avilamycin, avoparcin, gentamicin and narasin but resistant to tetracycline and tylosin and intermediately resistant to copper. Only Enterococcus gallinarum and Enterococcus casseliflavus were resistant to vancomycin and virginiamycin resistance was present in less than half the Enterococcus faecium isolates. Zinc resistance was largely confined to Enterococcus faecalis but bacitracin resistance was uncommon in E. faecalis in comparison with the other species. Tiamulin resistance was common in all species except E. casseliflavus. Resistance to flavophospholipol was detected in most E. faecium isolates and in a high proportion of E. gallinarum, E. casseliflavus and E. hirae/durans but was only found in one isolate of E. faecalis. No tetO, rplC, rplD, vanA, vanB, vatA and vatD genes were found. The presence of ermB, tetL, tetM, tcrB, aac6-aph2, tetK, tetS, vanC1, vanC2, lsaA, lsaB and vatE varied between the species and largely corresponded to the susceptibility phenotype. The findings show that resistance to antibiotics of high clinical significance for nosocomial Enterococcus infections is absent, whereas antimicrobial resistance was detected for some other antibiotics including bacitracin, flavophospholipol, tetracycline, tiamulin, tylosin and virginiamycin.     

Interaction between probiotic lactic acid bacteria and canine enteric pathogens: a risk factor for intestinal Enterococcus faecium colonization?

Selected probiotic lactic acid bacteria (LAB) have been shown to elicit positive health effects particularly in humans. Competitive exclusion of pathogens is one of the most important beneficial health claims of probiotic bacteria. The effect of probiotic LAB on competitive exclusion of pathogens has been demonstrated in humans, chicken and pigs. In this study we evaluated the ability of certain LAB strains (Lactobacillus rhamnosus GG, Bifidobacterium lactis Bb12, Lactobacillus pentosus UK1A, L. pentosus SK2A, Enterococcus faecium M74 and E. faecium SF273) to inhibit the adhesion of selected canine and zoonotic pathogens (Staphylococcus intermedius, Salmonella Typhimurium ATCC 14028, Clostridium perfringens and Campylobacter jejuni) to immobilised mucus isolated from canine jejunal chyme in vitro. Adhesion of C. perfringens was reduced significantly by all tested LAB strains, between 53.7 and 79.1% of the control without LAB, the LAB of canine origin yielding the best reduction. The adhesion of S. Typhimurium and S. intermedius were not significantly altered by any of the LAB included in the study. Both enterococci tested significantly enhanced the adhesion of C. jejuni, to 134.6 and 205.5% of the control without LAB. E. faecium may thus favor the adhesion and colonization of C. jejuni in the dog's intestine, making it a potential carrier and possibly a source for human infection. Enhanced C. jejuni adhesion is a new potential risk factor of enterococci. Our results further emphasize the importance of safety guidelines to be established for the probiotics intended for animal use.     

Screening of Staphylococcus aureus, Staphylococcus intermedius, and Staphylococcus schleiferi isolates obtained from small companion animals for antimicrobial resistance: a retrospective review of 749 isolates (2003-04)

Companion animal staphylococcal isolate antibiograms were screened retrospectively to determine the frequency of methicillin-resistant (MR) infection by Staphylococcus aureus, Staphylococcus intermedius, and Staphylococcus schleiferi. Rates of MR were: S. aureus 35%, S. intermedius 17%, and S. schleiferi 40%. Frequency of isolation of methicillin-resistant S. aureus (MRSA) from dogs and cats was similar, whereas methicillin-resistant S. intermedius (MRSI) and methicillin-resistant S. schleiferi (MRSS) were significantly more common in dogs. MRSS was more commonly associated with superficial (skin and ear canal) infections, whereas MRSA was more commonly associated with deep infections. The MR strain resistance pattern to other classes of antibiotics was also investigated. MRSA was resistant to the most classes of antibiotics, followed by MRSI, while MRSS maintained the most favourable susceptibility profile. MR staphylococci may pose a significant risk to animal and public health. Therefore, to avoid selecting for resistant strains in cases of suspected staphylococcal infection, clinicians should consider culture and susceptibility testing early in the course of treatment.     

Lactic acid bacteria for mink. Colonization and persistence of Enterococcus faecium Cernelle 68 in the digestive tract of mink.

A method was developed to follow a lactic acid bacterial strain, Enterococcus faecium Cernelle 68, with respect to adhesion, multiplication, colonization, and persistence in the digestive tract of mink. Also the spread of the strain in the cage was examined. When adding 5 x 10(9) c.f.u. of a rifampicin resistant mutant per kg feed, high viable counts were registered throughout the digestive tract, apart from the oesophagus. Counts were increasing in the aboral direction, suggesting some multiplication in the intestine. It was possible to detect the strain in the intestinal tract 4 days after discontinuation of administration. Neither culture nor scanning electron microscopy gave evidence to suggest that E. faecium Cernelle 68 adhered to the mucosa. The spread of the E. faecium strain was observed in the environment. Counts of E. coli, lactobacilli, staphylococci, and clostridia were low, and none of these bacteria were constant findings.     

Isolation，Identification and Antibiotics Resistance Analysis of Enterococcus from Yak

17 feces samples of yak which were collected in Hongyuan county were measured with Gram staining method and 16S rRNA molecular identification in this study.8 suspected Enterococcus were separate from feces samples by bacteria purification and PCR amplification with 1 500 bp specific band. 6Enterococcus faecalis and 2Enterococcus faecium were identified through 16S rRNA sequencing.The homology analysis of the strains revealed that the homology between Enterococcus faecalis and reference strains sequence were 99.7% to 100%,that of Enterococcus faecium and reference sequence were 98.2% to 99.2%,indicating that the yak Enterococcus was highly conserved in the process of genetic evolution.The drug sensitive test results showed that the isolated strains were highly resistance to aminoglycoside antibiotics.Enterococcus faecium 11-1-2 strain was not only 5 multi-resistant,but also showed resistence to vancomycin.Enterococcus faecalis strains was most 3 multi-resistant.The antibiotics resistance results revealed that the resistance of yak Enterococcus was serious and should be taken seriously.     

牦牛源肠球菌的分离鉴定及耐药性分析

试验结合革兰氏染色及16S rRNA分子鉴定,对分离自红原县的17份牦牛粪便样中的肠球菌进行鉴定。结果显示,通过细菌分离纯化及PCR扩增,从牦牛粪便样品中分离出8株疑似肠球菌,分离菌的扩增产物经凝胶电泳后均产生1 500 bp特异性条带。16S rRNA测序结果显示,8株疑似肠球菌中,6株为粪肠球菌,2株为屎肠球菌。同源性比对分析显示,粪肠球菌与参考序列同源性为99.7%~100.0%,屎肠球菌与参考序列同源性为98.2%~99.2%,说明牦牛源肠球菌在遗传进化过程中高度保守。运用K-B纸片法进行药敏试验,结果显示,分离株对氨基糖苷类抗生素耐受性较高,2株屎肠球菌中,11-1-2菌株表现为5重耐药,且该菌株耐万古霉素,粪肠球菌主要表现为3重耐药,药敏结果提示牦牛源肠球菌耐药较严重,应引起高度重视。     

Evaluation of the components of a commercial probiotic in gnotobiotic mice experimentally challenged with Salmonella enterica subsp. enterica ser. Typhimurium

Vitacanis((R)), a probiotic preparation containing a Lactobacillus acidophilus, an Enterococcus faecium and a Saccharomyces cerevisiae, has been developed for the prevention of intestinal disorders in dogs and cats. In the present study, these microorganisms were tested jointly or singly during experimental infection of gnotobiotic mice with Salmonella Typhimurium. Four experimental groups consisting of animals given probiotics jointly or singly and a control group consisting of germfree mice were used. The groups were treated with one or three of the microorganisms (experimental) or PBS (control) 10 days before intragastric challenge with a suspension containing about 10(2) cells of the bacterial pathogen. A higher survival (P<0.05) was observed in gnotobiotic mice given E. faecium (82%). All the animals in the other groups died after the challenge but the survival time was longer (P<0.05) for groups given all three of the microorganisms (7.4+/-2.4 days) or given only L. acidophilus (7.2+/-2.9 days) than for the control mice (4.4+/-1.1 days) and the mice that received S. cerevisiae (4.9+/-1.6 days) mice. The survival data agreed with the histopathological findings which showed more severe liver and intestinal lesions in control mice and in mice given Saccharomyces. In vitro antagonistic assays showed inhibition growth of E. faecium and S. Typhimurium around the colonies of L. acidophilus and for S. Typhimurium around the colonies of E. faecium. However, in vivo, S. Typhimurium became similarly established in the digestive tract of gnotobiotic mice at levels ranging from 10(8) to 10(10)CFU/g of feces and remained at these high levels until the animals died or were sacrificed. Among the three probiotic components of the commercial product Vitacanis((R)), E. faecium was the only one that provided protection against challenge with S. Typhimurium. Protection was not due to the reduction of the intestinal populations of the pathogenic bacteria.     

Feline salmonellosis. A nosocomial outbreak and experimental studies.

An outbreak of S. typhimurium gastroenteritis and septicemia is described in young cats admitted to a veterinary hospital for routine medical and surgical reasons. A morbidity of 32% and a mortality of 61% was observed in the outbreak. Affected cats exhibited oral shedding of the organism and contaminated feed and water dishes may have been the vehicle of spread of infection. Possible detrimental effects of incorrect choice of antibiotic therapy are discussed. Attempts to reproduce the disease in a group of laboratory cats were unsuccessful. Antibody production was poorly correlated with infection in these cats.     

猪源粪肠球菌和屎肠球菌多重PCR快速鉴定方法的建立

粪肠球菌和屎肠球菌是引起猪感染发病的优势肠球菌种,以肠球菌的16S rRNA基因设计属特异性引物,利用SodA基因多态性设计种特异性引物,同时优化反应条件,建立了能同时测定猪源粪肠球菌和屎肠球菌多重PCR方法。通过特异性和敏感性分析,该方法能扩增出肠球菌属特异性片段及粪肠球菌和屎肠球菌种特异性片段;对粪肠球菌敏感性为0...     

Monitoring of antimicrobial resistance in healthy dogs: first report of canine ampicillin-resistant Enterococcus faecium clonal complex 17

National surveillance programs on antimicrobial usage and antimicrobial resistance in animals have been established in various countries but few of them include bacteria from pets. The objectives of this study were to assess the prevalence of antimicrobial resistance in healthy dogs and to search for resistance phenotypes of clinical relevance. Escherichia coli and Enterococcus spp. were isolated from faecal swabs obtained from 127 dogs. Disk diffusion was used to measure antimicrobial susceptibility in 117 Escherichia coli, 10 Enterococcus faecium and 51 Enterococcus faecalis of canine origin. Resistance was relatively low compared with food animal species in Denmark. All Escherichia coli isolates were susceptible to broad-spectrum aminopenicillins, third generation cephalosporins and fluoroquinolones. Despite the low prevalence of resistance, statistical analysis of questionnaire data revealed a significant association (p=0.02) between recent antimicrobial treatment and resistance in Escherichia coli. Interestingly, two dogs were found to shed Enterococcus faecium resistant to ampicillin. Multilocus sequence typing of these isolates indicated that the two isolates belonged to sequence types associated with human nosocomial infections, and one (ST-192) was genetically related to human epidemic clonal complex 17. The detection of ampicillin-resistant Enterococcus faecium warrants further studies on the prevalence of these bacteria in dogs and on the possible implications to both animal and human health. The results suggest that distinct methods for detection and assessment of antimicrobial resistance in animals should be considered depending on the target animal species and the purposes of the study.     

产γ-氨基丁酸屎肠球菌的筛选及γ-氨基丁酸的定量

本试验旨在利用分子生物学方法鉴定分离得到1株产γ-氨基丁酸(GABA)屎肠球菌,并定量测定所产GABA的量。从泡菜、酸奶、土壤、新鲜牛奶样品中筛选出一目标菌株F6,进行形态学特征与革兰氏染色鉴定;再扩增菌株F6的16S r DNA基因,然后测定该基因序列和构建系统发育树;同时采用高效液相色谱(HPLC)法定量测定菌株F6发酵液中的GABA含量。结果表明:菌株F6菌落大而光滑、圆形、直径1~2 mm、边缘整齐、乳白色;在分离培养基上菌落周围形成透明圈,使分离培养基呈黄色;在MRS固体培养基上菌落不透明,周围有透明圈。革兰氏染色鉴定菌株F6为阳性菌。分子生物学鉴定分析显示,菌株F6的16S r DNA基因序列与Gen Bank数据库中屎肠球菌(Enterococcus faecium)的相似性大于99%。HPLC法测定得到GABA标准曲线线性方程为Y=7 080 733.139 5 X-4 511.692 7(R2=0.999 4),通过方程得出菌株F6发酵液中的GABA含量为7.1 g/L,保留时间为7~10 min。结果提示,本试验筛选得到1株高产GABA的屎肠球菌F6。     

Antimicrobial susceptibility of Enterococcus strains isolated from poultry faeces

We have investigated the resistance of Enterococcus isolated from poultry faeces to antibiotics commonly used as therapy of enterococcal infections. Identification was made by the method of Facklam and Collins. Minimal inhibitory concentrations of penicillin, ampicillin, vancomycin and teicoplanin were determined and high level aminoglycoside resistance was investigated. Genes codifying high level aminoglycoside resistance (HLAR) were determined by PCR. Fifty five Enterococcus strains were isolated (63.6% E. faecalis, 12.7% E. mundtii, 9.1% E. faecium, 7.3% E. casseliflavus, 3.7% E. durans and 3.6% E. hirae). None of the strains were resistant to VAN, TEC, P or AM. HLAR was found in 34.5% of strains for SM, 27.3% for KM and 7.3% for GM. The gene for the bifunctional enzyme was found only in one strain, that showed HLAR to GM and KM. Fourteen strains harboured the gene aph(3')-III, being 11 resistant to KM and STR, and three resistant to GM, KM and STR. The remaining six strains showed HLAR to STR, but were negative for the three genes tested by PCR. The gene ant(4'4") was not detected in any of the strains. No unexpected vancomycin resistance was detected. The resistance rates among poultry strains were lower than those found among human strains isolated from hospital patients in recent Canary studies.     

Role of coagulase-negative staphylococci in human disease

Coagulase-negative staphylococci (CNS) are normal inhabitants of human skin and mucous membranes. They have long been dismissed as culture contaminants, but now the potentially important role of CNS as pathogens and their increasing incidence has been recognized. Approximately 55-75% of nosocomial isolates is methicillin resistant. CNS were the first organisms in which glycopeptide resistance was recognized. In the immunocompetent host, CNS endocarditis and urinary tract infections with Staphylococcus saprophyticus are the most common CNS infections. Other patients are usually immunocompromised, with indwelling or implanted foreign bodies. CNS account for approximately 30% of all nosocomial blood stream infections. The majority of these concern catheter-related sepsis. Other important infections due to CNS include central nervous system shunt infections, endophthalmitis, surgical site infections, peritonitis in patients with continuous ambulatory peritoneal dialysis and foreign body infections. CNS are rarely associated with mastitis in humans. Staphylococcus lugdunensis is more pathogenic than other CNS as it expresses several potential virulence factors. The distinction between clinically significant, pathogenic and contaminating isolates is a major problem. Several studies show clonal intra and inter hospital spread of Staphylococcus epidermidis strains which suggests that infection control measures may be necessary for multiresistant CNS isolates as for methicillin resistant Staphylococcus aureus. As a result of medical progress, mainly due to the use of invasive and indwelling medical devices, CNS are now a major cause of nosocomial and health-care related infections.     

一株致病性粪肠球菌的分离与鉴定

从1羽发病雏鸡中分离到1株溶血性革兰氏阳性球菌。初步分离鉴定后经攻毒试验能够造成小白鼠和雏鸡死亡,具有致病性。药敏试验表明,该菌对多种药物均表现耐药性,仅对庆大霉素、青霉素、氧氟沙星敏感。通过BLAST软件分析该菌株的16S rRNA序列,确定该致病性细菌为粪肠球菌。     

In vitro antimicrobial activity of orbifloxacin against Staphylococcus intermedius isolates from canine skin and ear infections

The objective of the study was to evaluate the in vitro activity of orbifloxacin against Staphylococcus intermedius strains isolated in France from canine skin and ear infections. The minimum inhibitory concentrations (MICs) of orbifloxacin against 240 field S. intermedius isolates (69 skin and 171 ear isolates) ranged from 0.016 to 8 mg l(-1), with MIC50 and MIC90 equal to 0.5 and 1 mg l(-1), respectively. Only one strain, a pyoderma isolate was resistant (MIC=8 mg l(-1)). Orbifloxacin was tested at different concentrations for killing rate against five isolates obtained from pyoderma cases and against a reference strain (Staphylococcus aureus ATCC 29213). Orbifloxacin expressed a concentration-dependent bactericidal activity against the S. aureus reference strain, but a time-dependent bactericidal activity against S. intermedius. Orbifloxacin induced bactericidal effect against the S. intermedius strains tested with concentrations equal to or two times MIC.