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1.
In the present study four methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains isolated from a dog (n = 3) and the anterior nares of the dog owner (n = 1) were investigated by conventional and molecular methods. The species identity of the four S. pseudintermedius strains was confirmed by conventional methods, by PCR mediated amplification of S. intermedius/S. pseudintermedius specific segments of thermonuclease encoding gene nuc and by restriction fragment length polymorphism analysis of phosphoacetyltransferase encoding gene pta. Investigation of the four S. pseudintermedius for toxinogenic potential revealed that all four strains were positive for the exfoliative toxin encoding gene siet and the leukotoxin encoding genes lukS, lukF. The oxacillin and penicillin resistance of the four S. pseudintermedius strains could be determined by cultivation of the strains on oxacillin resistant screening agar base, ChromID MRSA Agar and Brilliance MRSA Agar and by multiplex PCR detecting the resistance genes mecA and blaZ. The genetic relatedness of the strains was studied by macrorestriction analysis of their chromosomal DNA using pulsed field gel electrophoresis (PFGE). According to PFGE all four S. pseudintermedius strains represent an identical bacterial clone indicating a cross transmission between the dog and the dog owner.  相似文献   

2.
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3.
Staphylococci isolated from animals (n=311) were screened for methicillin resistance by oxacillin agar screening. Oxacillin-resistant strains were tested for the presence of the mecA gene by PCR. Isolates were identified by standard techniques and 16S rDNA analysis, and their antimicrobial susceptibilities were tested using an agar diffusion method. MecA-positive strains were further analyzed using pulsed-field gel electrophoresis (PFGE). From 11 multidrug-resistant staphylococci, 6 were mecA-positive: 2 methicillin-resistant Staphylococcus aureus (MRSA) and 4 Staphylococcus haemolyticus. Screening of 300 staphylococci (100 S. aureus, 100 S. intermedius and 100 coagulase-negative staphylococci (CNS)) randomly chosen from the strain collection of the Veterinary Microbiological Diagnostic Center yielded five oxacillin-resistant coagulase-negative staphylococci, four of which were mecA-positive. PFGE showed that all mecA-positive staphylococci isolated from animals had distinct patterns. However, one MRSA isolated from a flank fistula of a dog showed homology to a human epidemic MRSA cluster, suggesting that transfer of MRSA between humans and dogs might occur.  相似文献   

4.
Methicillin/oxacillin resistance of 10 S. intermedius strains was investigated by conventional and molecular methods. The strains tested had been isolated in Germany during routine veterinary microbiological examinations of specimens from a small animal clinic between May and September 2005. Epidemiological relationships of the strains were studied by macrorestriction analysis of their chromosomal DNA using pulsed field gel electrophoresis (PFGE). Species identity of the 10 S. intermedius strains was confirmed by conventional methods and by PCR mediated amplification of S. intermedius specific segments of thermonuclease encoding gene nuc. As controls, four methicillin/oxacillin resistant S. intermedius (MRSI) strains obtained from specimens sent by four veterinarians and three selected methicillin/oxacillin sensitive S. intermedius (MSSI), also obtained from the small animal clinic, were tested. The 10 strains, representing approximately 6% of all S. intermedius isolated from the clinic throughout the time period mentioned above, and the four MRSI obtained from veterinarians, were methicillin/oxacillin and penicillin resistant using disk diffusion tests and could be cultivated on oxacillin resistant screening agar base (ORSAB). Both resistances could be confirmed by multiplex PCR detecting the resistance genes mecA and blaZ. The three MSSI were methicillin/oxacillin sensitive in all tests. Epidemiological investigation by macrorestriction analysis of the chromosomal DNA of the strains by pulsed field gel electrophoresis revealed that all 10 MRSI strains obtained from the clinic and the four MRSI strains obtained from veterinarians, in contrast to the three MSSI strains, represent identical or closely related bacterial clones possibly indicating a cross-infection of the animals in the clinic and the distribution of a single MRSI clone in the pet population.  相似文献   

5.
Using different typing methods (MLST, spa‐, SCCmec‐ and agr‐typing), PFGE and DNA microarray‐based chip analysis, we characterized 20 MRSA strains isolated from livestock and veterinarians. PFGE analysis after macrorestriction with EagI provided seven different band patterns, which could be grouped into four clusters. One cluster consisted of all MRSA ST398 strains isolated from pigs, calves, mastitis milk and two veterinarians. One strain of ST398 from a veterinarian and the two strains of ST1 and ST8 formed the three other clusters. Antimicrobial susceptibility testing showed that 15 of 20 strains were resistant to ampicillin, cefoxitin, clindamycin, erythromycin, oxacillin, penicillin and tetracycline. All strains were susceptible to rifampin and vancomycin, 19 were susceptible to ciprofloxacin and 18 were susceptible to sulphamethoxazole/trimethoprim. Genes encoding different enterotoxins, leukotoxins and haemolysins were found in certain strains.  相似文献   

6.
本研究旨在了解甘肃地区奶牛乳房炎金黄色葡萄球菌的耐药性和耐甲氧西林金黄色葡萄球菌的感染情况,为奶牛乳房炎的防制提供理论依据。采用KB纸片扩散法,检测17株金黄色葡萄球菌对8种不同抗菌药物的敏感性;再用琼脂稀释法检测了苯唑西林、万古霉素对金黄色葡萄球菌的最小抑菌浓度(MICs);头孢西丁纸片扩散法和PCR扩增特异性mecA耐药基因对所有受试菌株进行全面的耐甲氧西林金黄色葡萄球菌检测。结果表明,菌株对青霉素、磺胺异恶唑具有较强抗性,而对环丙沙星、头孢唑啉、万古霉素和苯唑西林全敏感;头孢西丁纸片扩散法未能检测出表型为MRSA的阳性菌株,而PCR方法却检测出8株mecA基因阳性菌株,且这些菌株的苯唑西林MIC均小于2 μg/mL。菌株的耐药情况较严重,对甲氧西林敏感而携带mecA基因的菌株高频存在于被调查地区的奶牛场中。  相似文献   

7.
The dog is the natural host of Staphylococcus pseudintermedius. Many research efforts are currently being undertaken to expand our knowledge and understanding of this important canine commensal and opportunistic pathogen. The objective of this review is to summarize the current knowledge of the species, including the latest research outcomes, with emphasis on taxonomy, diagnostics, ecology, epidemiology and pathogenicity. Despite the important taxonomic changes that have occurred over the past few years, the risk of misidentification in canine specimens is low and does not have serious consequences for clinical practice. Staphylococcus pseudintermedius carriage in the dog is more frequent and genetically heterogeneous compared with that of Staphylococcus aureus in man. It appears that these staphylococcal species have evolved separately through adaptation to their respective natural hosts and differ with regard to various aspects concerning ecology, population structure and evolution of antibiotic resistance. Further understanding of the ecology and epidemiology of S. pseudintermedius is hampered by the lack of a standard method for rapid and discriminatory typing and by the limited data available on longitudinal carriage and population structure of meticillin‐susceptible strains. With regard to pathogenicity, it is only now that we are starting to explore the virulence potential of S. pseudintermedius based on genomic and proteomic approaches, and more research is needed to assess the importance of individual virulence factors and the possible existence of hypervirulent strains.  相似文献   

8.
Although methicillin-resistant Staphylococcus aureus (MRSA) were generally isolated from human beings; these agents were recently isolated from various animal species. It has been shown that MRSA isolates are not only resistant to beta-lactam antibiotics, but can also be resistant to the other commonly used antibiotics. In this study, 18 phenotypic methicillin resistant S. aureus isolates from bovine mastitis cases were analyzed by PCR for the presence of mecA gene encoding methicillin resistance and aac(6′)/aph(2″), aph(3′)-IIIa and ant(4′)-Ia genes encoding aminoglycoside resistance. Out of 18 S. aureus isolates (oxacillin MICs, ≥4 μg/ml), 3 were positive for mecA gene. Only one from 3 mecA positive isolates was positive for genes encoding aminoglycoside-modifying enzymes and this isolate carried aac(6′)/aph(2″) in combination with aph(3′)-IIIa gene. The aph(3′)-IIIa gene was detected in 3 isolates. These three isolates carrying the aminoglycoside-modifying enzyme genes were resistant to gentamicin, kanamycin and neomycin. The mecA gene of 3 MRSA isolates was sequenced. All three mecA genes of these isolates were identical to that found in human MRSA strains, except a one-base substitution at nucleotide position 757. From the data presented in this study, it can be concluded that MRSA isolated from bovine mastitis may be originated from human beings, but further studies are needed to investigate the possibility of zoonotic transfer of MRSA.  相似文献   

9.
This is a case report of enteric protothecosis caused by Prototheca zopfii in an eight-year-old male mixed breed dog with a history of chronic bloody diarrhea, loss of appetite and weight loss. Algae were isolated from rectal scrapings in defibrinated sheep blood agar and dextrose Sabouraud agar. Cytological evaluation showed the presence of globular and cylindrical organisms with a defined capsule and variable number of endospores, characteristic of the genus Prototheca, in the rectum of the animal. Scanning electron microscopy of P. zopfii strains at different development stages confirmed the diagnosis of algal infection. Molecular identification using a conserved 18S rDNA gene sequence determined that the strain belonged to genotype 2. This report describes success on treatment of canine protothecosis, diagnosed based on clinical, cytological, microbiological, scanning electron microscopy and genotypical findings.  相似文献   

10.
The aim of this study was to investigate the phenotypic and genotypic diversity and anti‐microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north‐eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti‐microbial susceptibility by Kirby–Bauer disc diffusion method. Confirmation of methicillin‐resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP‐2a latex agglutination. Clonal relatedness of isolates was determined by pulsed‐field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase‐negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622‐ST1625). The findings show that MRSA is prevalent in milk from semi‐extensive dairy cows in north‐eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm‐to‐table continuum is warranted.  相似文献   

11.
Staphylococcus pseudintermedius is a commensal of dogs that is implicated in the pathogenesis of canine pyoderma. This study aimed to determine if S. pseudintermedius expresses surface proteins resembling those from Staphylococcus aureus and to characterise them. S. pseudintermedius strain 326 was shown to adhere strongly to purified fibrinogen, fibronectin and cytokeratin 10. It adhered to the α-chain of fibrinogen which, along with binding to cytokeratin 10, is the hallmark of clumping factor B of S. aureus, a surface protein that is in part responsible for colonisation of the human nares. Ligand-affinity blotting with cell-wall extracts demonstrated that S. pseudintermedius 326 expressed a cell-wall anchored fibronectin binding protein which recognised the N-terminal 29 kDa fragment. The ability to bind fibronectin is an important attribute of pathogenic S. aureus and is associated with the ability of S. aureus to colonise skin of human atopic dermatitis patients. S. pseudintermedius genomic DNA was probed with labelled DNA amplified from the serine-aspartate repeat encoding region of clfA of S. aureus. This probe hybridised to a single SpeI fragment of S. pseudintermedius DNA. In the cell-wall extract of S. pseudintermedius 326, a 180 kDa protein was discovered which bound to fibrinogen by ligand-affinity blotting and reacted in a Western blot with antibodies raised against the serine-aspartate repeat region of ClfA and the B-repeats of SdrD of S. aureus. It is proposed that this is an Sdr protein with B-repeats that has an A domain that binds to fibrinogen. Whether it is the same protein that binds cytokeratin 10 is not clear.  相似文献   

12.
Records of all Diagnostic laboratory submissions from 2012 to 2015 were examined and subjected to analysis according to species, location of infection, species of bacteria, and antibiotic resistance/susceptibility. A total of 23.8% of all culture isolates were Staphylococcus sp. Of those Staphylococcus, 43% were isolated from surgical site infections.Staphylococcus pseudintermedius accounted for approximately 28% of all staphylococcus cultures, while methicillin-resistant (MR) S. pseudintermedius accounted for 8% of all staphylococcus cultures. Environmental samples were also collected by swabbing surfaces in the intensive care unit (ICU) and anesthesia prep room at the OSU VTH. Isolated bacterial colonies were subjected to PCR for species identification and for the presence of the mecA gene. Ability of horizontal transfer in vitro of the mecA gene was evaluated by incubating the mecA positive bacterium, with the mecA negative bacterium, and then plated onto agar plates infused with known concentration of oxacillin. Colonies were then subjected to PCR for species and mecA identification. Horizontal transfer of the mecA gene was demonstrated and confirmed via PCR from MR S. epidermidis to MS S. pseudintermedius in an in vitro model that mimicked the veterinary hospital environment. Biofilms were established using four Staphylococcus species isolated from swabbing the Intensive Care Unit (ICU) and anesthesia prep room and were resistant when exposed to the current cleaning agent.Staphylococcus species makeup nearly ¼ of all infections at OSU VDL during the four years of the study, and MS S. pseudintermedius was shown to acquire the mecA gene from an environmental strain.  相似文献   

13.
Staphylococcus pseudintermedius is part of the normal canine flora but frequently causes pyoderma in canine atopic dermatitis (AD). This study aimed to determine whether particular S. pseudintermedius strains were associated with AD and/or pyoderma. Ninety‐six S. pseudintermedius isolates from the ear, nares, perineum and lesions of 21 atopic and 16 healthy dogs were lysed with proteinase K and digested with 40 U SmaI. Restriction products were separated using pulsed‐field gel electrophoresis (PFGE) with an Oxford S. aureus control and lambda‐ladder DNA concatomer markers. A dendrogram was constructed by the unweighted pair group method. All isolates showed a ≥56% similarity coefficient. Nine distinct PFGE clusters were identified, as follows: five from both atopic and healthy dogs; three from atopic dogs only; and one from healthy dogs only. Nine clusters were isolated from the nares, eight from the perineum, five from the ears and six from pyoderma lesions. There were no significant differences in the frequency of isolation from atopic or healthy skin, body sites or infected lesions for any of the clusters. Two of six healthy dogs and 18 of 20 atopic dogs with multiple isolates had closely related isolates (less than three band differences) at more than one sampling site. Isolates from pyoderma lesions were closely related to at least one mucosal isolate in 11 of 16 dogs. Staphylococcus pseudintermedius isolates appear to be heterogeneous, and colonization or infection of atopic skin was not associated with any particular strain or cluster of strains.  相似文献   

14.
Methicillin resistance mediated by the mecA gene in Staphylococcus aureus, also known as “true MRSA”, is typically associated with high oxacillin MIC values (≥8 mg/L). Because non-mecA-mediated oxacillin resistant S. aureus phenotypes can also cause hard-to-treat diseases in humans, their misidentification as methicillin-susceptible S. aureus strains (MSSA) can compromise the efficiency of the antimicrobial therapy. These strains have been refereed as Borderline Oxacillin-Resistant S. aureus (BORSA) but their characterization and role in clinical microbiology have been neglected. Considering the increasing importance of livestock-associated methicillin-resistant S. aureus ST398 (LA-MRSA) as an emerging zoonotic pathogen worldwide, this study aimed to report the genomic context of oxacillin resistance in porcine S. aureus ST398 strains. S. aureus isolates were recovered from asymptomatic pigs from three herds. Oxacillin MIC values ranged from 4 to 32 mg/L. MALDI-TOF-confirmed isolates were screened for mecA and mecC by PCR and genotyped by means of PFGE and Rep-PCR. Seven isolates were whole genome sequenced. None of the isolates harbored the mecA gene or its variants. Although all seven sequenced isolates belonged to one sequence type (ST398), two different spa types (t571 and t1471) were identified. All isolates harbored conserved blaZ gene operon and no mutations on genes encoding for penicillin-binding-proteins were detected. Genes conferring resistance against other drugs such as aminoglycosides, chloramphenicol, macrolide, lincosamide and streptogramin (MLS), tetracycline and trimethoprim were also detected. Isolates also harbored virulence genes encoding for adhesins (icaA; icaB; icaC; icaD; icaR), toxins (hlgA; hlgB; hlgC; luk-PV) and protease (aur). Pigs can serve as reservoirs of non-mecA-mediated oxacillin-resistant ST398 strains potentially pathogenic to humans. Considering that mecA has been the main target to screen methicillin-resistant staphylococci, the occurrence of BORSA phenotypes is probably underestimated in livestock.  相似文献   

15.
Since 2003 eleven Swiss sheep flocks were affected by abortion storms due to Salmonella abortusovis, an infection which had not been reported in this country for decades although cases of salmonellosis are notifiable in Switzerland. This raised doubts about the adequacy of the currently used diagnostic tools and the origin of this infection. Therefore, PCR was tested for its potential as a more rapid and more reliable method for diagnosing S. abortusovis infections under field conditions. Fecal and vaginal samples were collected at different times after abortion and PCR was used to detect bacterial DNA. Bacteria were isolated by conventional culture techniques. For determining their origin they were analyzed by pulsed field gel electrophoresis (PFGE) and compared to isolates from Germany and France. Sequencing of randomly selected amplicons allowed confirming the specificity of the result. PCR was more sensitive because it allowed detecting S. abortusovis DNA up to three months after infection even in samples that were negative by culture. Escherichia coli from the digestive tract of sheep could inhibit the growth of S. abortusovis in vitro suggesting that the lower sensitivity of diagnosis by bacterial culture may in part be due to growth inhibition of S. abortusovis by resident bacteria. Results of PFGE indicated that the Swiss strains were closely related among themselves but distinct from German and French strains suggesting the presence of an autochthonous infection.  相似文献   

16.
Recent reports suggest that methicillin-resistant strains of Staphylococcus schleiferi subspecies coagulans are now commonly isolated from dogs. Given the association of a potentially mobile SCCmec type IV element with lysogenic phage-encoded Panton Valentine Leukocidin (PVL) toxin genes in community-acquired methicillin-resistant Staphylococcus aureus strains we hypothesized that methicillin-resistant S. schleiferi ssp. coagulans strains may also encode PVL toxin genes. Forty S. schleiferi ssp. coagulans strains isolated from companion animals were studied. Susceptibility to oxacillin was determined by broth microdilution and all isolates were screened by PCR for the presence of the mecA gene. SCCmec typing was performed on 14 isolates. A real-time PCR assay was developed for the detection of the PVL genes using a SmartCycler. Pulsed-field gel electrophoresis (PFGE) was performed to determine whether S. schleiferi ssp. coagulans strains were homogeneous. Twenty-eight of the 40 isolates (70%) were resistant to oxacillin and 26/28 possessed the mecA gene by PCR. SCCmec IV was identified in seven strains; the other seven isolates were not typable by this technique. All 40 strains were negative for the PVL toxin gene. PFGE showed a heterogeneous population and 13 different profiles were determined. In conclusion, this study showed that PVL toxin genes were not detected in a heterogeneous population of methicillin-resistant S. schleiferi ssp. coagulans strains isolated from companion animals.  相似文献   

17.
Community‐associated methicillin‐resistant Staphylococcus aureus (MRSA) is a serious public health concern and in Australia, one that disproportionately affects Aboriginal people. Paralleling MRSA in human medicine, methicillin‐resistant S. pseudintermedius (MRSP) is an increasingly prevalent pathogen in veterinary medicine. We aimed to characterize the carriage of MRSA and MRSP in dogs and cats from predominantly Aboriginal communities in a very remote region of New South Wales (NSW), Australia. Pets (303 dogs and 80 cats) were recruited from six communities in western NSW. Three swabs were collected from each animal (anterior nares, oropharynx and perineum) and from skin lesions or wounds (if present) and cultured on selective media for methicillin‐resistant staphylococci. Human host‐adapted community‐associated MRSA representing four multilocus sequence types (ST1‐IV, ST5‐IV, ST72‐IV, ST93‐IV) were isolated from eight dogs (prevalence 2.6%, 95% confidence interval 1.3%–5.1%). Two ST5‐IV isolates from a single dog were phenotypically trimethoprim‐resistant, harbouring trimethoprim‐resistant gene dfrG within the SCCmec type IVo mobile genetic element. MRSA was not isolated from any cats and MRSP was not isolated from any dogs or cats. This study estimated a high prevalence of human host‐adapted community‐associated MRSA carriage in dogs despite an absence of MRSP. This suggests MRSA carried by dogs in remote NSW originate from human hosts. The cycle of transmission between people, dogs and common environmental sources warrants further investigation. To our knowledge, this is the first report of trimethoprim‐resistant ST5‐IV in eastern Australia and the first report of trimethoprim‐resistant ST5‐IV from a dog.  相似文献   

18.
A 4-year-old intact female American Pit Bull Terrier from Italy descendant of an American-born bitch was evaluated for anorexia, lethargy, weakness, and intermittent vomiting. On physical examination, the dog was dehydrated, had pale mucous membranes, hunched posture and abdominal pain. A moderate anemia was observed. Splenomegaly and hyperechoic regions suspected as infarcts in the spleen were seen on abdominal ultrasound. Based on the suspicion of splenic torsion, splenectomy was performed. After surgery, the clinical condition deteriorated. A follow-up complete blood count demonstrated severe macrocytic normochromic anemia with evidence of marked regeneration, left shift neutrophilia, monocytosis and marked thrombocytopenia. Blood smear evaluation revealed single to multiple, variable sized (1–3 μm in diameter), and round to oval to band-like piroplasms within many red blood cells consistent with small form Babesia spp. or Theileria spp. A partial segment of the 18S rRNA gene was amplified and the PCR product was analyzed by direct sequencing. The nucleotide sequence was completely identical to that of Babesia gibsoni present in GenBank®. This is the first molecular detection and characterization of B. gibsoni infection in a sick dog from Italy.  相似文献   

19.
CASE HISTORY: A 14-year-old neutered male Sealyham terrier was referred for assessment of a persistent pyoderma. It had experienced numerous episodes of dermatitis involving pododermatitis, pyoderma and otitis over the previous 6 years.

CLINICAL FINDINGS: Superficial, focally deep and mucocutaneous pyoderma were present, with yellow mucoid exudate on both nares and the lower lips crusted with haemopurulent exudate. Epidermal collarettes were present on the dorsal and lateral trunk. There were peri-anal crusts and mild erythema was present on the concave aspect of both pinnae.

MICROBIOLOGICAL FINDINGS: Culture and microbiological testing identified Staphylococcus pseudintermedius as the infecting organism. Kirby-Bauer disc susceptibility testing revealed the isolate was resistant to numerous antimicrobials including oxacillin. PCR testing of the isolate identified the presence of the mecA gene which confers resistance to β-lactam antimicrobials. Pulsed field gel electrophoresis typing suggested the isolate was not related to the methicillin-resistant S. pseudintermedius that had been reported to be associated with canine infections in Western Australia.

DIAGNOSIS: Superficial, deep and mucus membrane pyoderma associated with a multi-drug resistant S. pseudintermedius.

CLINICAL RELEVANCE: This is the first recorded case of canine pyoderma involving methicillin-resistant multidrug-resistant S. pseudintermedius in New Zealand. Treatment of such cases is difficult because the number of effective and available antimicrobials is limited. This finding should raise the awareness of the veterinary and medical professions to the presence of such organisms in New Zealand and stimulate a discussion about possible biosecurity barriers, treatment strategies and prevention of zoonotic and nosocomial infections.  相似文献   


20.
Staphylococcus pseudintermedius was implicated as the cause of rapidly progressive and fatal necrotizing fasciitis in a dog. The isolate was methicillin-susceptible and did not contain genes encoding the Panton-Valentine leukocidin. While Streptococcus canis is typically considered to be the main cause of necrotizing fasciitis in dogs, staphylococci should also be considered.  相似文献   

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