Staphylococcus hyicus in cattle

Staphylococcus hyicus (subsp hyicus) was found to be a frequently occurring inhabitant of the skin of cattle. Significant numbers of this bacterium were found to be present in mange lesions of cattle between one and three years old. Experimental inoculations revealed that S hyicus had a pathogenic effect on the superficially scarified skin of young cattle.     

Trends in antimicrobial susceptibility in relation to antimicrobial usage and presence of resistance genes in Staphylococcus hyicus isolated from exudative epidermitis in pigs

From 1996 to 2001 a total of 467 Staphylococcus hyicus isolates from exudative epidermitis (EE) in pigs in Denmark were examined for susceptibility to 13 different antimicrobial agents. The presence of selected genes encoding macrolide (erm(A), erm(B) and erm(C)), penicillin (blaZ), streptogramin (vat, vga, vga(B), vat(B), vat(D) and vat(E)), streptomycin (aadE) and tetracycline resistance (tet(K), tet(L), tet(M) and tet(O)) were determined in selected isolates.The occurrence of erythromycin resistance increased from 33% in 1996 to a maximum of 62% in 1997 and decreased to 26% in 2001. Resistance to sulphametazole increased from 17% in 1996 to 30% in 1998 but has since decreased to 4% in 2001. Resistance to trimethoprim increased to 51% in 1997 and decreased to 21% in 2001. Resistance to tetracycline (21-31%) remained relatively constant during 1996-2000, but increased to 47% in 2001. Resistance to penicillin (54-75%) streptomycin (33-53%) and tetracycline (21-47%) remained relatively constant over the time investigated.All 48 penicillin resistant isolates examined contained the blaZ gene and 40 (85%) of the streptomycin resistant isolates the aadE gene. It was not possible to detect any streptogramin resistance gene in four streptogramin resistant isolates. Of the 55 erythromycin resistant isolates examined, five contained erm(A), 13 erm(B), 35 erm(C) and two both erm(A) and erm(C). The presence of erm(B) was confirmed by hybridization to plasmid profiles in all 13 PCR-positive isolates. Of 52 tetracycline resistant isolates examined, two contained tet(L), 38 tet(K) and 12 both tet(K) and tet(L).     

Bacteriophage typing of Staphylococcus hyicus subsp hyicus isolated from pigs

Four phages were isolated and used for typing Staphylococcus hyicus subsp hyicus isolated from pigs with or without exudative epidermatitis (EE) in Japan. Sixty-four (85.3%) of the 75 isolates examined were typeable at either routine test dilution (RTD) or 100 X RTD. Two or more kinds of phage patterns were present in the isolates from each pig with EE. All isolates from healthy pigs showed a single-phage pattern. Fourteen (32.6%) of 43 isolates and 7 (87.5%) of 8 isolates from pigs with EE in Belgium and Czechoslovakia, respectively, were typeable with the 4 phages. None of 180 isolates of S aureus, 7 (6.4%) of 110 isolates of S intermedius, and 2 (2.3%) of 86 isolates of S epidermidis were typeable.     

Proteolytic zymograms of Staphylococcus hyicus subsp. hyicus isolated from pigs, chickens and cows

The extracellular proteases of Staphylococcus hyicus subsp. hyicus were assayed by a zymogram showing caseinolysis and gelatinolysis. Four bands were associated with caseinolysis or with gelatinolysis. The patterns shown by strains isolated from pigs, chickens and cows were compared; isolates from pigs differed from those isolated from chickens or cows but strains isolated from diseased and healthy pigs could not be differentiated.     

Plasmid-encoded antibiotic resistance in Staphylococcus hyicus

A total of 33 Staphylococcus hyicus-cultures from piglets with exudative epidermatitis were analyzed for the presence of antibiotic resistance plasmids. Four small plasmids encoding resistances to chloramphenicol, macrolide-lincosamide-antibiotics, streptomycin or tetracyclines could be identified in plasmid-curing and plasmid-transformation experiments. For further characterization these plasmids were digested with restriction endonucleases. This led to the construction of a specific restriction map for each of the 4 plasmids. On the basis of their restriction maps, these 4 antibiotic resistance plasmids from "porcine" S. hyicus-cultures were compared with the respective resistance plasmids of other staphylococcal species from infections of humans and animals.     

Staphylococcus hyicus virulence in relation to exudative epidermitis in pigs.

Staphylococcus hyicus strains with different phage types, plasmid profiles, and antibiotic resistance patterns were isolated from piglets with exudative epidermitis. The strains could be divided into virulent strains, producing exudative epidermitis, and avirulent strains, producing no dermal changes when injected in experimental piglets. The results showed that both virulent and avirulent strains were present simultaneously on diseased piglets. This constitutes a diagnostic problem. Concentrated culture supernatants from nine virulent strains injected in the skin of healthy piglets produced a crusting reaction in all piglets. Acanthosis was observed in the histopathological examination of the crustaceous skin. Concentrated culture supernatants from nine avirulent strains produced no macroscopic or microscopic skin changes. Protein profiles from all virulent strains and seven out of nine avirulent strains showed a high degree of protein band homology. An approximately 30 kDa protein present in all concentrated culture supernatants capable of producing skin changes, could not be detected in samples that did not produce skin changes. No other protein showed a similar association. It is concluded that crusting reaction of piglet skin is a suitable indicator of virulence in S. hyicus in relation to exudative epidermitis, and that virulent strains produce a 30 kDa protein, absent in concentrated culture supernatants from avirulent strains. This 30 kDa protein might be an exfoliative toxin.     

The sensitivity to antimicrobial agents of species of Yersinia isolated from cattle and pigs in Nigeria

Fourteen isolates of yersiniae belonging to three species isolated from cattle and pigs were tested for their sensitivity to 12 antimicrobial agents by the tube dilution technique. All the isolates were sensitive to gentamicin and 93.0%, 93.0%, 85.7% and 85.7% sensitive to tetracycline, nitrofurantoin, chloramphenicol and sulphamethoxazole-trimethoprim, respectively. There were no consistent differences in the rates and patterns of resistance based on source, species and serotypes of organisms.     

Importance of Staphylococcus hyicus ssp hyicus as a cause of arthritis in pigs up to 12 weeks of age

Lame pigs, up to 12 weeks of age, were necropsied to establish a diagnosis. Of 175 pigs examined, 165 were confirmed to have arthritis by histopathological examination of joint tissues. Lesions were most common in the elbow and tarsal joints and least common in the joints of the feet. Typically, there was severe fibrinopurulent inflammation of synovial membranes regardless of the bacteria isolated. A bacterial aetiology was found in 114 (69%) of the 165 pigs. In arthritic pigs in which an aetiology was established the causative agents were Staphylococcus hyicus ssp hyicus (24.6%), Streptococcus equisimilis (26.3%), Actinomyces pyogenes (13.2%), Staphylococcus aureus (7.9%) and Haemophilus parasuis (7.9%). While gender did not affect the prevalence of arthritis, there was an age influence, most of the pigs culled for arthritis being under 6 weeks of age.     

An investigation of exudative epidermitis (greasy pig disease) and antimicrobial resistance patterns of Staphylococcus hyicus and Staphylococcus aureus isolated from clinical cases

Exudative epidermitis (EE) is a common skin disease of young pigs, caused mainly by Staphylococcus hyicus. Increased prevalence of EE and poor response to treatment are reported. Common strategies used by Ontario pork producers to treat pigs with EE were determined using a survey. Injection of penicillin G was reported as the most common parenteral antibiotic choice. Antimicrobial resistance patterns of S. hyicus and Staphylococcus aureus isolated from clinical cases (30 herds with samples from approximately 6 pigs per farm) showed that 97% of S. hyicus isolates were resistant to penicillin G and ampicillin; 71% of these isolates were resistant to ceftiofur. Similar resistance was noted among S. aureus isolates. Antimicrobial resistance has become a problem in the treatment of EE in Ontario.     

Zinc-resistance gene czrC identified in methicillin-resistant Staphylococcus hyicus isolated from pigs with exudative epidermitis

Methicillin-resistant Staphylococcus hyicus (MRSH) was investigated for czrC, a gene conferring zinc-resistance. The czrC gene was identified in 50% (14/28) of MRSH isolates, representing 14 pigs with exudative epidermitis from 8 farms. Newly weaned pigs, which are particularly susceptible to exudative epidermitis, are commonly fed high levels of zinc oxide.     

Antimicrobial resistance in staphylococci from animals with particular reference to bovine Staphylococcus aureus, porcine Staphylococcus hyicus, and canine Staphylococcus intermedius.

Besides their role as commensals on the skin and mucosal surfaces, staphylococci may be involved in a wide variety of diseases in animals. Staphylococcal infections in animals are mainly treated with antimicrobial agents and as a consequence, staphylococci from animal sources have developed and/or acquired resistance to the respective antimicrobial agents. Resistance statistics obtained from national monitoring programmes on staphylococci from cattle and pigs, but also from surveillance studies on staphylococci involved in diseases in dogs are reported and reviewed with regard to their comparability. This review mainly focusses on the genetic basis of antimicrobial resistance in staphylococci of animal origin. Particular attention is paid to resistance to those antimicrobial agents which are most frequently used in veterinary medicine, but also to antimicrobial agents, such as chloramphenicol and mupirocin, which are used in specific cases for the control of staphylococcal infections in pets and companion animals. In addition, plasmids and transposons associated with the respective resistance properties and their ways of spreading between members of the same or different staphylococcal species, but also between staphylococci and other gram-positive bacteria, are described.     

Isolation and restriction endonuclease analysis of a tetracycline resistance plasmid from Staphylococcus hyicus

A plasmid of 4.550 kb, conferring resistance to tetracycline, was demonstrated in Staphylococcus hyicus cultures from piglets with exudative epidermidis. The plasmid-encoded properties were determined both by curing and interspecific protoplast transformation experiments. The tetracycline resistance (TET) plasmid, designated pST1, was characterized by restriction endonuclease analysis and a preliminary restriction map was constructed. The pST1 plasmid was demonstrated in 19 (57.6%) of 33 S. hyicus cultures by Southern blot hybridization. It was also detectable by electron microscopy.     

Antimicrobial resistance of fecal coliforms from pigs in a herd not exposed to antimicrobial agents for 126 months

Patterns of antimicrobial resistance were determined for lactose-fermenting fecal coliforms obtained during a 20-month period from pigs in a herd without exposure to antimicrobial drugs for 126 months. Mean percent resistance to cephalothin, chloramphenicol, kanamycin, naladixic acid, streptomycin and tetracycline was lower (P less than 0.01) for isolates obtained during the 20-month period (Group 2) than for isolates obtained during the first 13 months (Group 1) after the withdrawal of antibiotics. Mean tetracycline resistance was 40.5% for Group 2 isolates. Approximately 44% of the Group 2 isolates were resistant to the 10 antimicrobial agents compared with 74.9% for the Group 1 isolates (P less than 0.01). Multiple resistance was lower (P less than 0.01) in the Group 2 isolates than in the Group 1 isolates. Four resistance patterns (tetracycline, tetracycline-streptomycin, tetracycline-sulfisoxazole, tetracycline-streptomycin-sulfisoxazole) accounted for 74% of the resistant isolates.     

Prevalence and mechanism of antimicrobial resistance in Staphylococcus aureus isolates from diseased cattle, swine and chickens in Japan

Antimicrobial administration is essential for the control and treatment of diseases in animals, but the emergence and prevalence of antimicrobial-resistant Staphylococcus aureus is a significant concern during animal production. Here we investigated the antimicrobial susceptibility of S. aureus from diseased food-producing animals and molecularly characterized the methicillin-resistant and fluoroquinolone-resistant isolates. A total of 290 S. aureus isolates obtained from cattle (n=246), swine (n=16), and chickens (n=28) between 2003 and 2009 were examined for antimicrobial susceptibility against 9 antimicrobials using an agar dilution method. Resistance to penicillin (PC) was most frequently found (24.8%), followed by oxytetracycline (OTC, 10.0%), dihydrostreptomycin (4.1%), erythromycin (EM, 3.1%), enrofloxacin (ERFX, 2.1%), and kanamycin (1.7%). The PC resistance rate was significantly higher in swine than in cattle (P<0.01) and chickens (P<0.01). The resistance rates to OTC, EM and ERFX were significantly higher in swine and chickens than in cattle (P<0.05). Methicillin-resistant S. aureus (MRSA) was recovered from milk derived from a cow with mastitis in 2003; sequence type 8, SCCmec type IV and spa type t024. In the six ERFX-resistant strains isolated after 2003, amino acid substitutions in ParC with/without GyrA were detected. As the prevalence of MRSA and FQ-resistant S. aureus in the animals should be noticed, continuous monitoring is necessary to control resistance to clinically important antimicrobials in S. aureus from food-producing animals.     

Isolation of exfoliative toxin from Staphylococcus hyicus subsp. hyicus and its exfoliative activity in the piglet

Exfoliative toxin was isolated from the sterile cell-free filtrate of 24 h culture of Staphylococcus hyicus subsp. hyicus strain P-1. The partial purification of exfoliative toxin produced by S. hyicus (shET) was performed by precipitation with 50-80% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column and column chromatography on DEAE-cellulose. Partially purified shET (pp-shET) caused exfoliation in piglets at 8 to 12 h after intradermal or subcutaneous injection. However, heat-treated pp-shET did not cause exfoliation in piglets for up to 24 h after injection. On histopathological examination of the skin at 12 h after injection of pp-shET, an intraepidermal cleavage plane was shown between the stratum corneum and stratum granulosum and at the stratum granulosum.