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1.
Phenotypic properties and species-specific PCR tests based on the nuc gene of Staphylococcus intermedius and S. aureus, and a conserved region of 16S rDNA were used to identify 45 S. intermedius and four S. aureus isolated from samples of dogs during routine diagnostics. Four S. pseudintermedius strains used for control purposes reacted positively with the S. intermedius nuc PCR showing the close relationship between both species. Investigating the 45 S. intermedius and four S. pseudintermedius strains for the prevalence of the exfoliative toxin SIET encoding gene yielded the presence of the gene for 21 of the S. intermedius and two of the S. pseudintermedius strains. Partial sequencing of the toxin gene of a single S. intermedius strain and comparing this sequence with that obtained from GenBank revealed an almost complete identity. The presence of the exfoliative toxin gene could mainly be found among S. intermedius isolated from skin and wound infections and from otitis externa possibly indicating a role of this toxin for the clinical symptoms.  相似文献   

2.
AIMS: To use multilocus enzyme electrophoresis to determine the genetic structure of Staphylococcus intermedius from normal skin of dogs and those isolated from a variety of disease conditions and to distinguish clinically important strains in dogs. METHODOLOGY: The diversity amongst 129 isolates of S intermedius from the skin and mucosa of 32 healthy dogs and 120 isolates from diseased sites in 120 individual dogs was examined using multilocus enzyme electrophoresis. Associations among ETs were examined to determine the diversity of isolates. RESULTS: Twenty two ETs were distinguished comprising 21 containing isolates from diseased sites and 11 containing isolates from normal dogs. The majority of isolates (171 of 249; 69% were located in two ETs (ET1 and ET 4), that were not distinguishable phenotypically. ET 1 contained 94 isolates (54 isolates from healthy dogs and 40 isolates from diseased sites) and ET 4 contained 77 isolates (46 from healthy dogs and 31 isolates from diseased sites). Further, 77.5% of isolates from healthy dogs were present in ET 1 and ET 4 and 59% of isolates from diseased dogs belonged to the same two ETs. There was only a small difference in genetic diversity among isolates taken from healthy dogs (11 ETs; H = 0.182) and those isolates taken from clinical specimens from diseased dogs (21 ETs; H = 0.218). Of the 21 ETs from diseased sites, ET 16 contained all six isolates from Staphylococcal Scalded Skin Syndrome in racing Greyhounds. CONCLUSIONS: The small difference in genetic diversity between isolates from the skin and mucosa of healthy dogs and isolates from various diseases, as well as the presence of the majority of isolates in two ETs, is consistent with the role of S intermedius as an opportunistic pathogen. The confinement of all Staphylococcal Scalded Skin Syndrome isolates within one ET is confirmation of this entity as a distinct disease of dogs.  相似文献   

3.
The occurrence of antimicrobial-resistant Staphylococcus intermedius strains was investigated in 13 dogs affected by deep pyoderma, their owners and 13 individuals without daily contact with dogs (control group). A total of 90 canine and 33 human S. intermedius isolates were typed by pulsed field gel electrophoresis (PFGE) to determine their possible identity. The occurrence of S. intermedius in dog-owners was significantly higher compared with the control group (Fisher's exact test, P=0.03), with S. intermedius being detected in seven dog-owners and in one individual not exposed to dogs. The results of the PFGE analysis showed that six out of 13 (46%) owners carried strains identical to those isolated from their dogs. Strains detected in both dogs and humans were resistant up to five different antimicrobial classes, including penicillins, fusidic acid, macrolides/lincosamides, tetracycline and chloramphenicol. Based on the results of this study, owners of dogs affected by deep pyoderma often carry multiple antimicrobial-resistant strains of S. intermedius occurring in their dogs. Independent of the direction and modalities of transmission, this finding raises questions concerning the possible transfer of resistance genes from canine S. intermedius to human pathogenic staphylococci.  相似文献   

4.
125-I-IgG binding activities were observed with 15 (17%) of 90 S. intermedius isolates from dogs and 39 (95%) of 41 S. hyicus isolates from pigs. Binding activities were not detected with S. hyicus isolates from cows. The IgG binding proteins of 2 S. intermedius, 2 S. hyicus, and protein A from S. aureus Cowan I were isolated from their cell surfaces. The proteins precipitated with IgG preparations from human, rabbit, pig, dog and horse, but not with IgG from cow, mouse and chicken. This indicated that these IgG binding proteins could be classified as type I receptors. In addition, the isolated proteins from all 3 staphylococcal species precipitated with polyclonal chicken anti-protein A antiserum. SDS-PAGE, Western blotting and gel isoelectric focussing of the proteins revealed numerous bands in the 42,000 D range and acid isoelectric points. The isoelectric point of the isolated proteins from both S. intermedius cultures was slightly more acidic than those from S. hyicus and S. aureus. The present results indicate a close functional and antigenic similarity, if not identity, between IgG binding proteins of S. intermedius and S. hyicus, and protein A of S. aureus.  相似文献   

5.
An exfoliative toxin (SIET)-producing strain (D-52) of Staphylococcus intermedius derived from canine pyoderma did not possess large plasmids. Therefore, the gene coding for SIET was considered to be located on the chromosomal DNA. The SIET gene was cloned from the chromosomal DNA of S. intermedius and was expressed in Escherichia coli. The nucleotide sequence of the SIET gene consists of a coding region of 990 bp specifying a polypeptide of 330 amino acid residues, which included a putative 42-residue signal sequence.  相似文献   

6.
Protein A in Staphylococcus intermedius isolates from dogs and cats   总被引:1,自引:0,他引:1  
The presence and quantity of extracellular and cell-bound protein A of Staphylococcus intermedius isolates from dogs and cats were determined, using an enzyme-linked immunoglobulin-binding assay. Horseradish peroxidase-conjugated rabbit anti-bovine immunoglobulin G purified by affinity chromatography was reacted with whole cell and supernatant fractions of S intermedius (n = 139), a protein A-producing strain of S aureus, and a protein A-deficient strain of S epidermidis. Extracellular protein A was found in 118 (84.9%) of 139 isolates of S intermedius. Most (69/118; 58.5%) of these isolates produced greater than 0.2 micrograms of extracellular protein A/ml. Cell-bound protein A was found in 6 (4.3%) of 139 isolates. Only 1 of these isolates contained cell-bound protein A exclusively. The other 5 isolates produced significantly greater amounts of extracellular protein A than cell-bound protein A. Additionally, greater than 96% of extracellular protein A could be removed from supernatants by adsorption with agarose gel containing immunoglobulin G.  相似文献   

7.
The aim of this study was to compare the adherence of four strains of Staphylococcus intermedius and a single strain of Staphylococcus hominis to corneocytes from both normal dogs and dogs suffering from atopic dermatitis. Cells from the skin surface, corneocytes, were collected from 10 normal dogs and 10 dogs suffering from atopic dermatitis. Four strains of S. intermedius, three isolated from canine pyoderma skin lesions (strains A, B and C), and one isolated form from canine synovial membrane sample from a case of septic arthritis (strain D) were compared. S. hominis, which is not normally associated with canine disease, was also evaluated for its ability to adhere to canine corneocytes. S. hominis did not adhere to canine corneocytes. All four strains of S. intermedius adhered well to canine corneocytes collected from both normal and atopic dogs. All strains of S. intermedius showed statistically greater adherence to corneocytes collected from atopic dogs compared with those collected from normal dogs. It was concluded that the adherence assay employed here showed that S. hominis does not adhere to canine corneocytes, S. intermedius adheres preferentially to atopic corneocytes.  相似文献   

8.
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.  相似文献   

9.
Staphylococci were isolated from the external auditory meatus in 14 (48.3%) of 29 dogs affected with otitis externa (OE dogs) and 28 (68.3%) of 41 dogs without OE (non-OE dogs). Twenty-two OE isolates were identified as belonging to 12 species, and 42 non-OE isolates were identified as belonging to 13 species. The predominant species found in both OE and non-OE isolates were S. intermedius, and S. epidermidis. Thirty-eight (59.4%) of 64 isolates were resistant to one or more of the 17 antimicrobial agents tested. Resistance to PCG and ABPC was most frequent. S. schleiferi subsp. coagulans, a recent etiologic agent of canine OE, was isolated from OE and non-OE dogs. All of the 5 S. schleiferi subsp. coagulans isolates showed typical characteristics. No clear difference in the extracellular enzyme or toxin profiles, nor in the PFGE patterns, was demonstrated between the OE and non-OE isolates of S. schleiferi subsp. coagulans. A new PCR primer set specific for 16S rDNA was designed to identify strains of S. schleiferi subsp. coagulans. The amplified fragment was detected in all of the 5 isolates as well as the type strain GA 211 (=JCM 7470) and a reference strain GA 11, but was not detected in any strains of the related species, S. aureus, S. intermedius and S. hyicus. The PCR may allow a simple, rapid and precise identification of S. schleiferi subsp. coagulans, in addition to the standard tube test for free coagulase.  相似文献   

10.
Aspects of carriage of Staphylococcus intermedius were studied in three groups of dogs. In 150 normal dogs presenting for annual booster inoculations, carriage rates for S . intermedius in the anterior nares (36%) and on the anal ring (36%) were recorded. In 44 dogs presented for routine elective surgery, carriage of S . intermedius was higher on the caudal nares (41%) than on the anterior nares (34%), oropharynx (25%) or anal ring (30%) allowing an assessment of the accuracy of basing nasal carriage rates on swabbing the anterior nares alone. A new finding was that animals from multidog households had significantly higher ( P < 0.005) nasal and anal carriage rates than dogs from single-dog households. In 33 dogs with superficial pyoderma, carrier rates were similar to those reported by others and significantly higher than in the other two groups with regard to both nasal and anal carriage ( P < 0.05 and P < 0.005, respectively).  相似文献   

11.
Staphylococcal pyoderma is rarely contagious between dogs and humans, or humans and dogs. This study investigated the hypothesis that there are species differences in adherence of Staphylococcus intermedius (the most common isolate from dogs) and Staphylococcus aureus (the most common isolate from humans) to canine and human corneocytes. Sheets of corneocytes were collected from the ventral abdomen of 10 dogs and the medial forearm of 10 humans (all healthy and without any history or physical signs of skin disease) using double-sided tape. Staphylococcus intermedius from a case of canine bacterial pyoderma and a human strain of S. aureus were prepared in phosphate buffered saline (PBS) and applied in duplicate, respectively, to the canine and human corneocyte-covered tapes using PBS as negative control. After incubation, rinsing, and staining with crystal violet, quantification of the adherent bacteria was carried out blindly by computerized image analysis. Staphylococcus intermedius was found to adhere significantly more to canine corneocytes than S. aureus (P = 0.0006), whereas S. aureus showed greater adherence to human corneocytes than S. intermedius (P < 0.0001). In addition, the pattern of adherence differed between the two organisms, with S. intermedius adhering to the entire surface and S. aureus adhering mainly to the periphery of both canine and human corneocytes. Preference for adherence to these two hosts may explain, in part, why S. intermedius and S. aureus are uncommonly isolated from human and canine skin infections, respectively.  相似文献   

12.
Temporal study of staphylococcal species on healthy dogs   总被引:5,自引:0,他引:5  
During a 1-year period, specimens were obtained monthly from 5 hair coat and 7 mucous membrane sites of 11 healthy dogs. Among 804 isolates of staphylococci, 13 species were identified. Staphylococcus intermedius was the most frequently isolated (40.2% of total isolates) coagulase-positive species, and S xylosus was the most frequently isolated (17.3%) coagulase-negative species. Moreover, S intermedius was the most frequently isolated species from the 12 sites evaluated and was isolated persistently from 8 of the 9 dogs that completed the 1-year study. On the basis of a commercial identification system, 14 profile numbers were identified for isolates of S intermedius. However, 2 profile numbers accounted for a majority (70.9%) of the isolates. Specific S intermedius biotypes identified on the basis of hemolysis, coagulase production, beta-lactamase activity, and antimicrobial susceptibility patterns were found repeatedly in 3 dogs. Seemingly, S intermedius was a resident of the normal bacterial microflora of these dogs; however, the inability to isolate S intermedius from 1 dog during the study year indicated that not all dogs harbor S intermedius as a resident microorganism.  相似文献   

13.
Staphylococcal pyoderma occurs commonly in atopic dogs. Some studies have suggested that adherence of staphylococci to corneocytes of atopic dogs and humans is higher than to corneocytes of healthy individuals. This hypothesis and possible differences resulting from the presence or absence of pyoderma, the severity of pruritus or the effect of treatment or gender, were studied. Adherent bacteria (Staphylococcus intermedius) were quantified by computerized image analysis on corneocytes collected from healthy or atopic dogs using double-sided adhesive tape. The adherence of S. intermedius to the corneocytes of atopic dogs was significantly greater than to those of healthy dogs (P=0.005). Furthermore, adherence was significantly greater in dogs with high levels of pruritus compared to those with low scores. No significant differences were found between atopic dogs with no history of pyoderma, atopic dogs with a history of pyoderma and atopic dogs with pyoderma at the time of sampling (P=0.068), suggesting that factors other than adherence are necessary for clinical pyoderma to develop. Treatment did not generally influence the adherence of S. intermedius to corneocytes of atopic dogs and there was no gender difference in adherence in either healthy or atopic dogs.  相似文献   

14.
Staphylococcus intermedius from pigeons, dogs, foxes, mink, and horses, was characterized by pulsed-field gel electrophoresis (PFGE) to evaluate the use of this typing method for discriminating among strains. SmaI cut the chromosomal DNA into 7-13 fragments ranging from approximately 48 kb to 655 kb, with most of the detectable fragments being smaller than 172 kb. S. intermedius from various animals had a high degree of restriction fragment length polymorphism. Pigeon strains have a similar genotype, despite the difference in their isolation area. Phage typing indicated that the dog, fox, and mink strains belong to the canine I or canine II type. The PFGE method further differentiated the mink strains from the dog and fox strains with regard to three fragments between 256 kb and 570 kb. As such, genomic DNA fingerprinting by PFGE appears to be an effective technique for discriminating S. intermedius strains from various animals. A combination of PFGE typing and phage typing would provide more detailed information than the single method for ecological investigations of S. intermedius.  相似文献   

15.
16.
Staphylococcus chromogenes is closely related to Staphylococcus hyicus, which is recognised as the causative agent of exudative epidermitis (EE) in pigs. S. chromogenes is part of the normal skin flora of pigs, cattle and poultry and has so far been considered non-pathogenic to pigs. A strain of S. chromogenes producing exfoliative toxin type B, ExhB, was identified by the use of a multiplex PCR specific for the exfoliative toxins from S. hyicus. The exfoliative toxin from S. chromogenes reacted in immunoblot analysis with polyclonal and monoclonal antibodies specific to ExhB from S. hyicus and had an apparent molecular weight of 30 kDa. Sequencing the gene encoding the exfoliative toxin from S. chromogenes revealed that the molecular weight of the toxin with the signal peptide and the mature toxin was 30,553 and 26,694 Da, respectively. Comparison of the exhB genes from S. chromogenes strain VA654 and S. hyicus strain 1289D-88 showed differences in seven base pairs of the DNA sequences and in two amino acid residues in the deduced amino acid sequences. Pigs were experimentally inoculated with S. chromogenes strain VA654. By clinical observations and histopathological evaluation of the skin alterations, all pigs revealed development of generalized exudative epidermitis. No toxin producing S. hyicus was isolated from the pigs and all ExhB-positive bacterial isolates were identified as S. chromogenes. This confirmed that the disease-causing agent was the inoculated S. chromogenes strain VA654. The results of this study show that S. chromogenes may cause exudative epidermitis in pigs.  相似文献   

17.
In piglets inoculated with partially purified exfoliative toxin (pp-shET) produced by Staphylococcus hyicus subsp. hyicus, exfoliation was observed at 12 h after injection. Chickens inoculated with the same dose of pp-shET also showed exfoliation within 30 min of injection. However, exfoliation was not demonstrated in mouse, rat, guinea pig, hamster, dog or cat inoculated with pp-shET until 24 h after injection. In cultured cell lines, especially L-929 and Hep-2, the rounding effect occurred after incubation with pp-shET for 1 h. The rounding effect was also seen in five other cultured cells (NCTC 2544, HeLa/S3, HmLu-1, CHO and BHK-21) 6-24 h after exposure to pp-shET. These round cells survived for 72 h after inoculation and formed a monolayer 24 h after changeover to a toxin-free medium. The rounding effect was observed in cells after the formation of the monolayer, but not before. It was suggested that the rounding effect was not caused by the increase in cyclic AMP in cells inoculated with pp-shET but by the cleavage of intracellular contacts.  相似文献   

18.
Staphylococcal colonization was compared in healthy dogs and in dogs with atopic dermatitis. Bacterial swabs were collected from the nasal mucosa, ear and perineum of 43 healthy and 24 atopic dogs and also from potentially infected skin lesions of the atopic dogs. Coagulase positive staphylococcal isolates were identified to the species level. At the time of this study Staphylococcus intermedius was considered a single species but has since been recognized as comprising at least three species with canine isolates believed to belong to Staphylococcus pseudintermedius . Of atopic dogs, 87.5% were colonized with S. intermedius compared to only 37.2% of healthy dogs. The ear was the only carriage site that showed any significant difference in S. intermedius isolation between healthy and atopic dogs. The perineum represented the most frequently colonized mucosal site for both groups. Sampling the nasal mucosa alone identified 71.4% of atopic and 37.5% of healthy S. intermedius carriers. Inclusion of a perineal swab identified 100% of atopic and 93.8% of healthy carriers. S. intermedius was isolated from all the lesional sites sampled from atopic dogs. Significantly fewer dogs were colonized by Staphylococcus aureus than S. intermedius , and there was no significant difference between S. aureus colonization of atopic and healthy dogs. S. aureus was not recovered from any lesions in atopic dogs. The results show that S. intermedius carriage is more prevalent in atopic dogs compared to healthy dogs and that to identify staphylococcal carriers both the nasal mucosa and the perineum should be sampled.  相似文献   

19.
We conducted an epidemiological study of Staphylococcus intermedius using arbitrarily primed PCR (AP-PCR) and antibiograms. One hundred and twenty-five S. intermedius isolates were recovered from the oral cavity and/or cranial hair coat of healthy dogs enrolled in a pet therapy program. Commensal S. intermedius was cultured from 32% of the oral cavity cultures and 13% of the cranial hair coat cultures. We characterized the colonization of the dogs as transient, intermittent, or persistent. For dogs characterized as persistently colonized, 73% of the isolates came from the oral cavity. These isolates were also genotyped by AP-PCR. A single major AP-PCR type was observed in 91% of the dogs (n=22); minor variations were frequently observed in these major types. Antibiograms of these commensal isolates were compared to antibiograms from 97 historical clinical isolates (1988-1992) obtained from cases of canine pyoderma. Resistance was most often observed to penicillin (64% and 55%) and tetracycline (38% and 38%) among the commensal and clinical isolates, respectively. The commensal isolates were significantly less resistant to erythromycin, clarithromycin, clindamycin, and trimethoprim/sulfamethoxazole. Our data suggests that differences in both genotype and antimicrobial susceptibility phenotypes exist among S. intermedius strains isolated from different anatomic sites from the same dog and supports the opportunistic nature of S. intermedius in canine infections.  相似文献   

20.
Over a year swabs were taken from 87 untreated bite wounds in dogs seen by veterinary practitioners in Harare, Zimbabwe. Swabs were also taken from normal skin adjacent to the wound site, and gingival swabs were collected from normal dogs coming to the same clinics. The swabs were cultured aerobically for pathogens, particularly Staphylococcus intermedius, and the antibiotic sensitivities of the pathogens were determined by disc diffusion assay. The most common pathogens isolated from the wounds were S intermedius (23 per cent), Escherichia coli (18 per cent) and non-lactose-fermenting coliforms (14 per cent). S intermedius was common on the normal skin of the dogs with infected wounds, and was associated with wounds on the abdomen, hindlimbs and tail and wounds that were more than three days old. This organism was, however, isolated only infrequently from the gums and there was little correlation in general between the prevalence of pathogens in the mouth and their prevalence in wounds. Of the S intermedius isolates from wounds, 30 per cent were resistant to penicillin and multiple antibiotic resistance was common among the enterobacterial isolates. The majority of the pathogens were sensitive to cotrimoxazole.  相似文献   

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