Frequency and antimicrobial susceptibility of Staphylococcus species isolated from canine pyodermas

Specimens obtained from pyogenic skin lesions of 210 dogs were culturally examined for staphylococci. A total of 215 isolates of staphylococci were biotyped, using the biochemical tests contained in a commercial staphylococcal identification system. Of 201 coagulase-positive isolates, 197 were identified as Staphylococcus intermedius, 3 as S aureus, and 1 as S hyicus. Of 14 coagulase-negative isolates, 5 were identified as S epidermidis, 5 as S xylosus, 3 as S simulans, and 1 as S hominis. Antimicrobial susceptibility tests were done on all staphylococcal isolates, using the standard disk-diffusion method. Staphylococcus intermedius isolates were susceptible to cephalothin, methicillin, and gentamicin. Resistance to ampicillin, penicillin G, and tetracycline was frequent. Antibiotic resistance was not associated with the depth of skin infection. Resistance to ampicillin, penicillin, tetracycline, and trimethoprim-sulfamethoxazole was not associated with previous antibiotic use. Increased resistance to chloramphenicol, clindamycin, and erythromycin was associated with previous antibiotic therapy. Antimicrobial susceptibilities of the other Staphylococcus species isolated are reported, but the small numbers of these species precluded making meaningful comparison with S intermedius.     

Antimicrobial resistance of feline staphylococci in south-eastern England

Staphylococcus aureus is reported as the predominant feline staphylococcal pathogen. There is concern that cats may transfer resistant staphylococci to humans. In this study, staphylococci were obtained from skin and mucosae of 20 domestic cats, 9 with lesions, and 10 healthy feral cats. Species were identified by DNase and API ID32 Staph tests. Of 187 isolates, 21.4% were coagulase-positive and predominately from lesional cats; 90% of these were Staphylococcus intermedius . Coagulase-negative species were isolated equally in all three groups. All isolates were susceptible to coamoxiclav, cephalexin and bacitracin. Twenty-two, including 18 coagulase-negative isolates, showed some resistance to cotrimoxazole, lincomycin, enrofloxacin or oxytetracycline. Two isolates were resistant to more than one antibiotic. More resistant isolates were obtained from feral cats ( P < 0.01). The results suggest that S. intermedius is the principal coagulase-positive species. Antibiotic resistance is generally low amongst feline staphylococci. Higher resistance amongst feral cats suggests exposure to environmental antibiotics.     

Evaluation of clinical characteristics and bacterial isolates in dogs with bacterial keratitis: 97 cases (1993-2003)

OBJECTIVE: To evaluate clinical characteristics and breeds affected with bacterial keratitis and compare patterns of resistance in bacterial isolates over time in dogs. DESIGN: Retrospective cross-sectional study. ANIMALS: 97 dogs with bacterial keratitis. PROCEDURE: Dogs with bacterial keratitis were identified from teaching hospital medical records at the Universities of Tennessee and Florida during the years 1993 to 2003. Data were collected pertaining to breed, Schirmer tear test results, treatments administered at the time of initial examination, bacterial species isolated, and resistance to selected antimicrobials. RESULTS: 66% of the dogs were brachycephalic, 54% had tear production < 15 mm/min, and 29% were receiving a corticosteroid at the time of initial examination. The most common bacteria isolated were Staphylococcus intermedius (29%), beta-hemolytic Streptococcus spp (17%), and Pseudomonas aeruginosa (21%). Staphylococcus intermedius isolates had limited resistance to certain antimicrobials. More than 80% of beta-hemolytic Streptococcus spp isolates were resistant to neomycin, polymyxin B, and tobramycin. Isolates of P aeruginosa were susceptible to tobramycin and gentamicin and had limited resistance to ciprofloxacin and enrofloxacin. Among bacterial species isolated, there was no evidence of development of antimicrobial resistance over time. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggested that administration of ciprofloxacin or a combination of a first-generation cephalosporin and tobramycin may be used in the treatment of bacterial keratitis while awaiting results of bacterial culture and susceptibility testing. Evidence suggests that current methods of medical management of bacterial keratitis are not associated with increased antimicrobial resistance.     

Pathogenic bacteria and fungi associated with external ocular diseases in dogs: 131 cases (1981-1986)

Medical records of 131 dogs with external ocular diseases were reviewed. Bacteriologic culture of swab specimens from 151 eyes revealed 100 eyes (66.2%) were considered positive for potentially pathogenic microorganisms. Of 127 species of microorganisms (bacterial and fungal) isolated, 50 (39.3%) were Staphylococcus spp (S intermedius, 17.3%). Streptococcus spp were the next most frequently isolated organism at 32 (25.2%), (Str canis, 16.5%). beta-Hemolytic streptococci (17%) were isolated more frequently than were alpha-hemolytic streptococci (9%), and coagulase-positive staphylococcal species (29%) were isolated almost 3 times as often as were coagulase-negative species (11%). Fungal and yeast organisms were isolated from 4.6% of the eyes. In vitro, most Staphylococcus spp were susceptible to cephalothin, bacitracin, and gentamicin, whereas most Streptococcus spp were susceptible to chloramphenicol, erythromycin, carbenicillin, and cephalothin. Pseudomonas spp were sensitive to tobramycin, gentamicin, and amikacin.     

Temporal study of staphylococcal species on healthy dogs

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.     

Genotypic relatedness of staphylococcal strains isolated from pustules and carriage sites in dogs with superficial bacterial folliculitis

OBJECTIVE: To determine whether staphylococcal isolates cultured from pustules and carriage sites in dogs with superficial bacterial folliculitis were genotypically the same strain by use of pulsed-field gel electrophoresis (PFGE). ANIMALS: 40 dogs with superficial bacterial folliculitis. PROCEDURES: Samples were obtained from 3 pustules and 3 carriage sites (anus, axillary skin, and nasal mucosa). Bacterial culture, morphologic identification, Gram staining, catalase and coagulase tests, speciation, and PFGE were performed. RESULTS: Of 246 isolates, 203 were Staphylococcus intermedius, 5 were Staphylococcus aureus, 15 were Staphylococcusspp, and 22 were coagulase-negative staphylococcal isolates. No dog had an isolate with the same PFGE pattern as an isolate from another dog. Coagulase-positive isolates from multiple pustules and multiple carriage sites had the same PFGE pattern in 37 of 39 (94.9%) and 22 of 39 (56.4%) dogs, respectively. Coagulase-positive staphylococcal isolates from at least 1 pustule had the same PFGE pattern as an isolate from at least 1 carriage site in 34 of 36 (94.4%) dogs. Ninety-seven of 116 (83.6%) coagulase-positive staphylococcal isolates from pustules had the same PFGE pattern as an isolate from at least 1 carriage site. Sixty-nine of 91 (75.8%) coagulase-positive staphylococcal isolates from carriage sites had the same PFGE pattern as an isolate from at least 1 pustule. CONCLUSIONS AND CLINICAL RELEVANCE: Coagulasepositive staphylococcal strains were heterogeneous among dogs with superficial bacterial folliculitis. In individual dogs, strains from multiple pustules were genotypically the same, and strains from pustules were genotypically the same as strains from carriage sites.     

Long-term study of aerobic bacteria of the genital tract in breeding bitches.

The aerobic bacterial flora of the genital tract was characterized in 59 bitches in an 18-month study. The bitches represented 4 breeds and were from 3 kennels. Collection of vaginal swab specimens for bacterial culturing was performed every month, except during estrus when specimens were collected every week (n = 826). The capsule of the swab containing transport media was broken before specimen collection to moisten the tip, which helped to reduce the number of negative cultures. All bitches helped at least once during the study and, thus, had known reproductive functions. Pregnancy rates, litter sizes, and pup mortality were within normal limits. Pasteurella multocida, beta-hemolytic streptococci group G, and Escherichia coli were the most common bacteria isolated. Although these species generally were isolated from mixed cultures, pure cultures were obtained from 18% of the specimens. There was a tendency for the various breeds to differ in their vaginal bacterial flora. The flora also varied during the reproductive cycle. Pasteurella multocida was isolated significantly more often during proestrus, estrus, metestrus, and pregnancy, than during anestrus and the postpartum period, and beta-hemolytic streptococci were isolated significantly more often during proestrus than during estrus, pregnancy, or the postpartum period. Staphylococcus intermedius was almost exclusively found after parturition. Culture results were negative for only 5.2% of specimens cultured. On the basis of our findings, bacterial culturing of vaginal swab specimens from bitches without signs of genital disease is of little value.     

Frequency and antimicrobial susceptibility of Staphylococcus spp isolated from feline skin lesions

Swab specimens obtained from skin lesions of 45 cats were cultured bacteriologically for staphylococci. Thirty-two staphylococcal isolates were recovered from 30 cats and were biotyped, using biochemical tests contained in a staphylococcal identification system. Of 23 isolates considered coagulase-positive, 16 were identified as Staphylococcus aureus, 5 as S intermedius, and 2 as S hyicus. Of 9 isolates considered coagulase-negative, 6 were identified as S simulans, 2 as S epidermidis, and 1 as S xylosus. Antimicrobial susceptibility tests were done on all staphylococcal isolates, using a disk-diffusion method. Staphylococcal isolates were susceptible to clavulanic acid-amoxicillin, cloxacillin, cephalothin, chloramphenicol, gentamicin, erythromycin, and trimethoprim-sulfamethoxazole. Resistance to penicillin G, ampicillin, and tetracycline was frequent.     

What’s happened to Staphylococcus intermedius? Taxonomic revision and emergence of multi‐drug resistance

Staphylococcus intermedius has been the predominant coagulase-positive Staphylococcus isolated from canine skin and mucosae and the most commonly reported staphylococcal pathogen in small animal practice for the last 35 years. Although microbiological tests have historically indicated variability in biochemical characteristics amongst S. intermedius isolates from animals, an acceptable level of diagnostic accuracy for clinical purposes was readily achievable with routine phenotypic testing. However, three recent developments have changed our understanding of the term "S. intermedius" and have challenged veterinary bacteriologists to ensure correct species identification of pathogenic staphylococci from small animals. First, the increasing recognition of meticillin-resistant Staphylococcus aureus in small animal practice and its human health implications demand accurate species identification. Secondly, the application of molecular techniques to analysis of staphylococcal isolates has led to a revised taxonomy and canine isolates of S. intermedius being re-named S. pseudintermedius. Thirdly, the recent, rapid emergence of meticillin- and multi-drug-resistant strains of Staphylococcus pseudintermedius (MRSP) has become a major therapeutic challenge in veterinary practice worldwide, including the UK. This article discusses the background of the recent taxonomic changes within the genus Staphylococcus and reviews the key features of MRSP and its implications for day-to-day laboratory diagnosis and small animal practice.     

Staphylococcal colonization of mucosal and lesional skin sites in atopic and healthy dogs

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.     

Identification of protein A from Staphylococcus intermedius isolated from canine skin

Protein A was identified in cell wall-bound and secreted forms from Staphylococcus intermedius isolated from canine skin. A direct binding radioimmunoassay for the detection of bacterial surface Fc receptors identified 48 of 50 S intermedius isolates that contained cell wall-bound protein A. Using a competitive binding radioimmunoassay for the detection of Fc-reactive proteins in bacterial culture supernatants, we identified 9 of 50 clinical isolates of S intermedius that secreted measurable quantities of an Fc receptor into the culture medium. Concentrated culture supernatants from these isolates were analyzed by western blotting techniques and probed with either a radiolabeled human IgG Fc-specific probe or a radiolabeled affinity-purified chicken antibody against protein A. The studies reported here confirmed that Fc receptors are secreted by S intermedius isolates from dogs and are antigenically and functionally similar or are identical to staphylococcal protein A. Analysis of Fc receptor secretion by S intermedius strains, isolated from dogs with a variety of dermatologic conditions, suggested a trend between severity of skin disease and the extent of Fc receptor secretion.     

CAMP-reaction among skin isolates obtained from a dog with an acute squamous eczema

The primary culture of a clinical specimen obtained from a dog with an acute squamous eczema revealed 3 different bacterial cultures. Two of these cultures, a beta-hemolytic Staphylococcus aureus and a group B streptococcal culture, demonstrated synergistic hemolytic activities on this primary culture plate. The group B streptococcus had the serotype surface antigens Ib/c, protein antigen c in its c beta component.     

Mustelidae are natural hosts of Staphylococcus delphini group A

According to the current taxonomy, the Staphylococcus intermedius group (SIG) comprises of at least three distinct species. While S. intermedius and S. pseudintermedius are associated with specific hosts (pigeons and dogs, respectively), the natural host of S. delphini remains unclear. We analysed 158 SIG isolates from less studied animal species belonging to the order Carnivora, including mink (n=118), fox (n=33), badger (n=6) and ferret (n=1). Species identification was performed by nuc PCR in combination with sodA sequence analysis and pta PCR restriction fragment length polymorphism (RFLP). The results showed a consistent association between host and bacterial species. All isolates from minks, ferret and badgers belonged to S. delphini group A, whereas all fox isolates except one were identified as S. pseudintermedius. The remaining fox isolate belonged to S. delphini group A. The results indicate that Mustelidae such as minks, ferrets and badgers are natural hosts of S. delphini group A. This is in contrast with Canidae, which are primarily colonized and infected with S. pseudintermedius. These findings suggest that coagulase-positive staphylococcal species may have evolved and diverged through host adaptation.     

Udder infection of goats by coagulase-negative staphylococci

Infection of udder halves by coagulase-negative staphylococci in seven commercial goat herds was studied in conjunction with the California Mastitis Test (CMT). Nine different species were identified and only 10% of strains belonged to groups which could not be identified with any of the known Staphylococcus species. The most prevalent species were Staphylococcus epidermidis (47.7%) and Staphylococcus caprae (19.7%). About half od the coagulase-negative staphylococcal infections gave negative CMT scores. The score was independent of the species of staphylococci involved and the stage of lactation. About 60% of the coagulase-negative staphylococcal species isolated were reisolated in the identical half udder during the following lactation.     

Methicillin resistance of staphylococci isolated from the skin of dogs with pyoderma

OBJECTIVE: To determine the methicillin-resistant profile of staphylococcal isolates from the skin of dogs with pyoderma. ANIMALS: 90 dogs with pyoderma. PROCEDURE: Staphylococci isolated from dogs with pyoderma were tested for susceptibility to methicillin by use of a standard disk diffusion test with oxacillin disks. The DNA extracted from the isolates was tested for the mecA gene that encodes the penicillin-binding protein 2a (PBP2a) by use of a polymerase chain reaction (PCR) assay. The expression of PBP2a was determined with a commercial latex agglutination assay. Species of staphylococcal isolates were identified by use of morphologic, biochemical, and enzymatic tests. RESULTS: Most of the isolated staphylococci were methicillin-susceptible, coagulase-positive Staphylococcus intermedius isolates. Whereas only 2 of 57 S. intermedius isolates were resistant to methicillin, approximately half of the isolates had the mecA gene and produced PBP2a. Staphylococcus schleiferi was the second most common isolate. Widespread resistance to methicillin was found among S. schleiferi isolates. More coagulase-negative S. schleiferi isolates were identified with mecA gene-mediated resistance to methicillin, compared with coagulase-positive S. schleiferi isolates. CONCLUSIONS AND CLINICAL RELEVANCE: The latex agglutination assay for the detection of PBP2a expression coupled with the PCR assay for the mecA gene may provide new information about emerging antimicrobial resistance among staphylococcal isolates.     

Prevalence of oxacillin- and multidrug-resistant staphylococci in clinical samples from dogs: 1,772 samples (2001-2005)

OBJECTIVE: To determine whether resistance to oxacillin and other antimicrobials in 3 Staphylococcus spp commonly isolated from dogs increased from 2001 to 2005. DESIGN: Retrospective case series. SAMPLE POPULATION: 1,772 clinical samples of various types obtained from dogs examined at the University of Tennessee Veterinary Teaching Hospital or at regional veterinary hospitals and submitted to the bacteriology and mycology laboratories associated with the teaching hospital. PROCEDURES: Samples were submitted by attending veterinarians to the bacteriology and mycology laboratories for routine aerobic microbial culture. Identification and antimicrobial susceptibility procedures were performed on all isolates. Susceptibility reports for each antimicrobial and Staphylococcus spp were determined from aggregate electronically archived test results. Oxacillin and multidrug resistance for Staphylococcus intermedius was analyzed by reviewing disk diffusion zone measurements. RESULTS: Oxacillin resistance increased among S. intermedius isolates during the past 5 years, and the increase was associated with multidrug resistance. In 2005, 1 in 5 Staphylococcus spp isolates from canine clinical samples was resistant to oxacillin. The most common staphylococcal species isolated were S. intermedius (n = 37), Staphylococcus schleiferi (21), and Staphylococcus aureus (4), and frequencies of oxacillin resistance in isolates of these species were 15.6%, 46.6%, and 23.5%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Veterinarians should be aware of the potential for empiric drug treatment failures in instances where Staphylococcus spp infections are common (eg, pyoderma). Judicious use of bacterial culture and susceptibility testing is recommended.     

Prevalence and Identity of Translocating Bacteria in Healthy Dogs

Bacterial translocation is characterized by the passage of intestinally derived bacteria across the intestinal mucosa to local or regional tissues. This phenomenon is believed to be important in the pathogenesis of gram-negative bacteremia and septicemia; however, the pathway or route of translocation remains unclear. To define the route of translocation better, mesenteric lymph nodes from 50 apparently healthy dogs undergoing elective ovariohysterectomies were cultured aerobically and anaerobically. The aims of this study were to determine the prevalence of bacterial translocation and to quantify and identify types of organisms found in mesenteric lymph nodes. Peripheral blood and portal blood samples were similarly cultured to rule out hematogenous organisms as a source of lymph node contamination. Bacteria were isolated from mesenteric lymph nodes of 26 dogs (52%). The number of bacteria varied from 50 to > 10⁵ organ-isms/g of tissue. Bacteria isolated included Staphylococcus intermedius (n = 3), coagulase-negative Staphylococcus (n = 2), nonhemolytic Streptococcus (n = 4), Bacillus species (n = 5), Escherichia coli (n = 6), Salmonella species (n = 3), Pseudomonas species (n = 2), Enterococcus species (n = 2), Clostridium sordelli (n = 1), Micrococcus species (n = 1), Lactobacillus species (n = 1), and Propionibacterium acnes (n = 1). One of 50 peripheral blood samples yielded an unidentified gram-positive coccus and a coagulase-negative Staphylococcus. No bacteria were isolated from portal blood samples of any dog. Further studies of this type on sick dogs are warranted before clinical recommendations can be made to culture mesenteric lymph nodes routinely.     

Hyaluronidase test in the diagnosis of staphylococci

Using the Streptococcus equi decapsulation test, 879 strains of S. aureus, 212 strains of S. intermedius and 214 coagulase-negative staphylococci were examined for hyaluronidase production; six of the coagulase-negative staphylococci belonged to S. hyicus subsp. hyicus. Positive hyaluronidase production was recorded in all strains of S. aureus and in the strains of S. hyicus subsp. hyicus. No hyaluronidase was produced by 208 coagulase-negative staphylococci and the strains of S. intermedius. The decapsulation test is recommended as a reliable method of the differentiation of S. aureus and S. intermedius.     

Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP

The coagulase-negative staphylococcal species causing mastitis in lactating cattle were identified and possible differences in the clinical characteristics or persistence of mastitis caused by different CNS were evaluated. The effect of antimicrobial treatment was also assessed. In addition, AFLP-typing of CNS was compared with the phenotypic identification. A total of 133 clinical or subclinical quarter cases of intramammary infection caused by CNS from the practice area of the Ambulatory Clinic of the University of Helsinki were studied. Bacteriological diagnosis was based on biochemical (API) testing. Staphylococcus simulans (43.6%) followed by S. chromogenes (23.3%) were the most common CNS species isolated from the milk samples. Ninety-nine isolates were genotyped using AFLP-analysis. Only 75.0% of S. chromogenes and S. simulans isolates identified with API test were clustered with the type strains of these species. Approximately half of the mastitis cases were clinical, and in the majority clinical signs were mild. The severity and persistence of intramammary infection were unaffected by CNS species. Fifty-nine percent of the quarter cases were treated with antimicrobials, and the rest were left without treatment. Mastitis due to beta-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were beta-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45.5% for untreated quarters. Bacterial cure rates for the most common CNS species or AFLP clusters were not statistically different. Further studies on identification of CNS species are needed.     

Vaccination against staphylococcal mastitis

Extract

Sir:- Immunisation with staphylococcal vaccines is a potential means of increasing the resistance to bacterial invasion of the udder. However, the development of an effective vaccine for control of staphylococcal mastitis in ruminants has proved to be an elusive goal. There have been numerous attempts to develop vaccines against coagulase-positive Staphylococcus aureus mastitis, but very few attempts to develop vaccines using coagulase-negative staphylococci. Development of a vaccine against coagulase-negative staphylococci may help control the sub-clinical mastitis caused by these organisms and assist in the search for an effective vaccine against mastitis caused by the more pathogenic coagulase-positive S. aureus.We describe here an experiment in which ewes were immunised with live coagulase-negative staphylococcal vaccine and challenged during lactation by intramammary infusion of a homologous strain.