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.     

Interaction of Staphylococcus hyicus and an inhibitor producing strain of Staphylococcus chromogenes on the skin of gnotobiotic piglets

Abstract— The effects of an inhibitor-producing strain of Staphylococcus chromogenes on the colonisation and disease produced by virulent and avirulent strains of Staphylococcus hyicus were examined. In the presence of S. chromogenes the colonisation of the virulent strain was reduced one thousand fold and the onset of lesions was delayed by 7 days. The ability of the avirulent strain of S. hyicus to colonise skin was greatly reduced and populations became almost undetectable within 6 days of inoculation. Résumeé— Les effets d'une souche de Staphylococcus chromogenes, productrice d'un facteur d'inhibition, ont été examinés sur la colonisation et la maladie produites par des souches non virulentes et virulentes de Staphylococcus hyicus. En présence de S. chromogenes, la colonisation de la souche virulente a été réduite de 1000 fois, et l'apparition des lésions a été retardée de 7 jours. La faculté pour la souche non virulente de S. hyicus de coloniser la peau a été considérablement réduite et les populations sont devenues à peu près introuvables 6 jours après l'inoculation. Zusammenfassung— Die Wirkungen Inhibitor-produzierenden Art von Staphylococcus chromogenes auf die Kolonisierung und Erkrankung der Haut durch virulente und avirulente Arten von Staphylococcus hyicus wurden untersucht. In Gegenwart von S. chromogenes wurde die Kolonisierung der virulenten Art um den Faktor 1000 verringert und des Auftreten der Hautveränderungen um 7 Tage verzögert. Die Fähigkeit zur Kolonisierung der Haut durch die avirulente 5. hyicus-Art wurde sehr stark verringert, ihre Population konnte innerhalb von 6 Tagen nach Inokulation nicht mehr nachgewiesen werden. Resumen Se estudian los efectos de una cepa de Staphylococcus chromogenes productora de un inhibidor sobre la colonización y la enfermedad producida por cepas virulentas y avirulentas de Staphylococcus hyicus. En presencia de S. chromogenes la colonización de cepas virulentas se redujo a una milésima parte y la aparición de lesiones se retrasó 7 días. La capacidad de cepas avirulentas de S. hyicus para colonizar la piel se vió fuertemente reducida y las poblaciones bacterianas eran prácticamente no detectables a los 6 días de la inoculación.     

Studies on the Virulence of Staphylococcus hyicus

Abstract— Culture supernatants from six Staphylococcus hyicus isolates were concentrated by ultrafiltration and were injected intradermally into the abdominal skin of 2-week-old piglets. Two distinct types of reaction were observed; (1) a focal erythema and (2) an exfoliative reaction with crusting. The severity of these reactions and the number of animals affected differed between isolates. The exfoliative reaction observed in these skin tests may be a good indicator of virulence. Résumé— Les surnageants de six souches de Staphylococcus hyicus ont été concentrés par ultrafiltration et furent injectés par voie intradermique dans la peau de l'abdomen de porcelets ägés de deux semaines. Deux types distincts de réactions furent observés: (1) un éryhème focal et (2) une réaction d'exfoliation avec apparition de croûtes. L'intensité de ces réactions et le nombre d'animaux atteints différaient selon les souches. La réaction d'exfoliation observée lors de ces tests pourrait être un bon indicateur de virulence. Zusammenfassung— Überstände der Kultur von sechs Staphylococcus hyicus-Isolaten wurden durch Ultra-zentrifugieren konzentriert und bei 2 Wochen alten Ferkeln intradermal in die Abdominalhaut injiziert. Es konnten zwei deutlich unterscheidbare Reaktionen beobachtet werden: (1) ein fokales Erythem und (2) eine exfoliative Reaktion mit Krustenbildung. Die Schwere dieser Reaktionen und die Zahl der betroffenen Tiere war je nach Isolat unterscheidlich. Die exfoliative Reaktion bei diesen Hauttests könnte ein guter Indikator für die Virulenz sein. Resumen Los sobrenadantes de 6 cultivos diferentes de Staphylococcus hyicus se concentraron mediante ultrafiltración y se inyectaron via intradérmica en la piel abdominal de lechones de dos semanas de edad. Se observaron dos tipos de reacción: (1) un eritem focal y (2) una reacción exfoliativa y costrosa. La gravedad de las reacciones y el nümero de animales afectados variaba en functión del cultivo utilizado. La reacción exfoliativa que se observó en estos tests intradérmicos puede ser un buen indicador de virulencia.     

Distribution of the putative virulence factor encoding gene sheta in Staphylococcus hyicus strains of various origins

In the present study, Staphylococcus (S.) hyicus strains isolated in Russia (n = 23) and Germany (n = 17) were investigated for the prevalence of the previously described genes sheta and shetb. Sheta was detected in 16 S. hyicus strains. Sheta-positive strains were mainly found among strains isolated from exudative epidermitis, and frequently together with the exfoliative toxin-encoding genes exhD and exhC. Partial sequencing of sheta in a single S. hyicus strain revealed an almost complete match with the sheta sequence obtained from GenBank. None of the S. hyicus strains displayed a positive reaction with the shetb-specific oligonucleotide primer used in the present study. According to the present results, the exotoxin encoding gene sheta seems to be distributed among S. hyicus strains in Russia and Germany. The toxigenic potential of this exotoxin, which does not have the classical structure of a staphylococcal exfoliative toxin, remains to be elucidated.     

A fatal suppurative pneumonia in piglets caused by a pathogenic coagulase-positive strain of <Emphasis Type="Italic">Staphylococcus hyicus</Emphasis>

Staphylococcus hyicus is one of the opportunistic pathogens that cause infections to animals. Early studies have demonstrated that S. hyicus is the causative agents of exudative epidermitis in pigs, arthritis in horses and chicken, mastitis in cow, and bacteremia, sepsis and multiple organ failure in humans. Here, we report the isolation and identification of a representative S. hyicus isolate, named JLHN15, from a pig farm with a disease characterized by bacteremia, suppurative pneumonia and fibrinous pericarditis. Our results indicate that JLHN15 is a pathogenic coagulase-positive Staphylococcus. To the best knowledge, this is the first report of S. hyicus causing an infection characterized by suppurative pneumonia and sepsis.     

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.     

The Ability of the Enzyme‐Linked Immunosorbent Assay to Detect Antibody against Staphylococcus aureus in Milk following Experimental Intramammary Infection

Changes in the milk antibody levels against Staphylococcus aureus were measured at the start of an experimental intramammary instillation of either S. aureus (Study I) or Staphylococcus hyicus (Study II). A commercial enzyme‐linked immunosorbent assay system was used. Twenty‐one Holstein cows were enrolled in Study I and 15 Holstein cows were used in Study II. Pathogen instillation began 21 days before the start of the non‐lactating period. Cows received intramammary antibiotic treatment in all quarters immediately after the last milking, the start of the non‐lactating period. Lacteal secretions were collected before the start of the non‐lactating period, and during the immediate postpartum period in both studies, and during the non‐lactating period in Study I. Milk was cultured for mastitis pathogens and S. aureus antibody levels and somatic cell counts were determined from all samples. There was an approximate 2‐week delay in the elevation in antibody levels in response to the instillation of S. aureus. Antibody levels remained elevated in cows with S. aureus intramammary infections postpartum, but were below threshold in cows where intramammary infections were cured during the non‐lactating period. Antibody levels were elevated by S. hyicus intramammary infections, remained elevated for the first 12 days postpartum, but were below threshold by day 21 postpartum. Cows with incipient intramammary S. aureus infections might be misclassified as false negatives by the antibody test. However, results suggest that cows with S. hyicus intramammary infections that were not cured would not be misclassified if milk is withheld from test for the first 30 days postpartum, as recommended by the manufacturer of the test.     

Exudative Epidermitis in Pigs. Bacteriological Studies on the Causative Agent Staphylococcus Hyicus

The properties of six pathogenic and six non-pathogenic strains of staphylococci from clinical cases of exudative epidermitis are described. The pathogenic strains were shown to produce typical lesions following experimental inoculation of pigs. All of the strains studied were classified among the staphylococci due to their ability to produce acid from glucose under anaerobic conditions. One of the six pathogenic strains was a slow coagulase producer. It is suggested that the species designation Staphylococcus hyicus be retained.     

Strains of Staphylococcus aureus and pathology associated with chronic suppurative mastitis in rabbits

Staphylococcal mastitis is one of the main reasons for culling adult does from commercial rabbitries. The aim of this study was describe the spectrum of gross and microscopic lesions in 178 cases of chronic staphylococcal mastitis in adult does and to determine whether there is a correlation between Staphylococcus aureus genotypes and pathology. On the basis of histopathology, chronic mastitis was differentiated into abscesses (66.3%), suppurative mastitis with a lobular pattern (7.9%), cellulitis (19.6%) and mixed lesions (6.2%). Pathological presentations were not related to S. aureus genotype.     

Assay of protein A in Staphylococcus hyicus subsp. hyicus by ELISA and immunoelectron microscopy

The presence and quantity of protein A in Staphylococcus hyicus subsp. hyicus isolates were examined by an enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy. Cell-bound protein A was demonstrated in 45 (94%) of 48 isolates from diseased pigs and in 113 (86%) of 132 isolates from healthy pigs by ELISA using peroxidase-conjugated rabbit antibody, but was not found in isolates from chickens and cows. Most of these swine isolates contained about 100 to 300 ng of cell-bound protein A/ml. Extracellular protein A was not detected in any isolates from pigs, chickens or cows. In the immunoelectron microscopy assay, swine isolates were labeled with goat anti-mouse IgG conjugated to colloidal gold particles, but chicken and cow isolates were not labeled.     

Characterization of Methicillin‐Resistant Staphylococcus aureus Isolates from Pig Carcasses in Hong Kong

This study describes the isolation and characterization of methicillin‐resistant Staphylococcus aureus (MRSA) from slaughtered pigs sampled from local markets in Hong Kong. The nares of 400 slaughtered pigs were cultured and MRSA isolates characterized for the presence of antibiotic‐resistance determinants, toxins and SCCmec and spa types using PCR. Clonality was investigated using PFGE and MLST. The prevalence of MRSA colonization of slaughter pigs was 39.3%, the majority (92%) harbouring SCCmec type IVb. Of the 157 samples yielding MRSA, 13 had two distinct MRSA strains present. Spa type t899 was predominant, with only 5/170 isolates displaying closely related types (t4474, t1939, t2922 and t5390). PFGE with sma1 and MLST confirmed the strains as ST9. Most isolates were multidrug resistant. Tetracycline resistance (97%) was mainly attributable to tet(K) with only 3% of isolates additionally harbouring tet(M). Resistance to erythromycin (89%) and chloramphenicol (71%) was associated with the presence of erm(C), and fex( A), respectively. No strains carried cfr and there was no resistance to linezolid, although minimum inhibitory concentration (MICs) were close to the resistance break point. Resistance to clindamycin (99%), ciprofloxacin(78%), quinopristin–dalfopristin (44%) and cotrimoxazole (32%) was common, but remained low for fusidic acid (4%) and rifampicin (2%). All strains were negative for PVL, exfoliative, and enterotoxins. This survey confirmed the uniformity of MRSA isolates in pigs from several regions of China, in contrast to more diversified characteristics reported in European studies. Colonization rates were higher than previously reported. Isolates were resistant to a wide range of antibiotics, but resistance was not detected to linezolid, nitrofurantoin, vancomycin or tigecycline. Although the clinical importance of ST9 in humans is uncertain, continued surveillance, in particular of those occupationally‐exposed, is recommended.     

Zinc Oxide Therapy Increases Prevalence and Persistence of Methicillin‐Resistant Staphylococcus aureus in Pigs: A Randomized Controlled Trial

There is concern that therapeutic use of zinc oxide (ZnO) in swine production may select for methicillin‐resistant Staphylococcus aureus (MRSA) due to co‐location of the zinc resistance gene (czrC) and methicillin resistance gene (mecA) within the staphylococcal cassette chromosome mec (SCCmec). The objective of this investigation was to determine whether MRSA carriage in pigs is influenced by exposure to therapeutic doses of in‐feed ZnO (3000 mg/kg) when compared to the recommended dietary levels (100 mg/kg). A randomized controlled trial was completed using 110 pigs that were naturally colonized with czrC‐positive MRSA. The pigs were followed from birth to weaning (21 d), at which point they were randomized into 8 pens and exposed to either a control feed (100 mg ZnO/kg feed; n = 49 pigs) or a treatment feed (3000 mg ZnO/kg feed; n = 50 pigs); neither feed contained additional antimicrobials. MRSA carriage was monitored weekly in each group for 4 weeks post‐weaning. The prevalence of MRSA was significantly higher in the treatment group at 1‐week (OR = 18.1; P < 0.01) and 2 weeks (OR = 3.01; P = 0.01) post‐weaning when compared to the control group, but there was no difference later in the nursery phase. Persistent MRSA carriage (testing positive ≥2 times post‐weaning) was observed in 2% (1/49) of control pigs and 22% (11/50) of treated pigs (P < 0.01). All MRSA isolates (spa types t034 and t3075) carried czrC and showed uniform resistance to zinc. These findings demonstrate that the prevalence and persistence of MRSA in nursery pigs can be affected by high levels of in‐feed ZnO in the absence of antibiotics.     

Progression of Mycoplasma hyosynoviae infection in three pig herds. Development of tonsillar carrier state, arthritis and antibodies in serum and synovial fluid in pigs from birth to slaughter

In this investigation, natural infection with Mycoplasma hyosynoviae was followed in groups of individual pigs in three different herds with regard to occurrence of tonsillar carrier state, clinical arthritis and development of antibodies in serum and in synovial fluid. Antibodies were detected by a polyclonal enzyme-linked immunosorbent assay (ELISA) developed for experimental use. The infection with M. hyosynoviae progressed very differently in the three herds investigated. In one herd, the infection was apparently limited to adult pigs. In a second herd, all pigs became tonsillar carriers of M. hyosynoviae, but no mycoplasma-related arthritis nor any serological response was demonstrated within the growing-finishing period. In the third herd investigated, tonsillar infection was detected in all pigs, clinical cases of M. hyosynoviae arthritis followed and a moderate serological response was observed in some, but not all, pigs. In all three herds, M. hyosynoviae infection was carried in the tonsils of the adult pigs, but it was only occasionally transmitted from sows to piglets. Maternal antibodies were transferred to the piglets and persisted for approximately 8-12 weeks. After weaning, some pigs became infected before 20 weeks of age, while others did not. In the majority of cases, the tonsillar infection was established from 11 weeks of age or older. A latent tonsillar infection was present for a period of several weeks within the group of investigated pigs before cases of generalized infection and arthritis were seen. In some cases, generalization of M. hyosynoviae infection in the blood and in joints was observed in spite of the detection of an active serological response a few weeks earlier. The present work suggests that generalization of the infection and development of arthritis may depend on age, immunity, virulence factors and/or infection pressure; in some herds maybe combined with certain triggering mechanisms such as stress and lowered general resistance.     

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.     

Production of enterotoxin(s) by Staphylococcus hyicus

Five strains of Staphylococcus hyicus (3 of subspecies hyicus and 2 of chromogenes) were tested serologically for their ability to produce known and unknown enterotoxin(s) and were also examined using monkey bioassays. None of the 5 strains produced any detectable level of the known enterotoxins A—E. However, all of them produced emetic responses in 2 or more of 6 cynomologus monkeys when culture growth was fed intragastrically. One of the 5 strains (S. hyicus subspecies hyicus, VII 76) produced emetic responses in 3 of 6 monkeys with 50 ml of culture growth. The other 4 strains required 1 l of culture broth (concentrated 20-fold) to produce an emetic response in at least 2 of 6 monkeys. Enterotoxin production is, therefore, not unique to S. aureus species. Since some of these organisms do not produce coagulase and thermonuclease, they would be ignored in food hazard evaluation.     

An evaluation of nonsuppurative joint disease in slaughter pigs

Fifty-two joints from pigs with nonsuppurative joint disease from a local abattoir were examined grossly, histologically, and microbiologically in order to establish macroscopic differences between degenerative arthropathy and arthritis due to an infectious organism. The joints were grouped grossly according to the type and severity of lesions of the synovial membrane and cartilage, and microscopically according to the severity of synovial membrane lesions. Osteochondrosis and Erysipelothrix rhusiopathiae were the most common causes of nonsuppurative joint disease in the joints examined. The major macroscopic differences between these two arthropathies were in the nature and severity of the synovial and cartilaginous lesions and involvement of the lymph node draining the diseased joint. Typically, in osteochondrosis, the changes are feathery hypertrophy of villi, focal full-thickness cartilage buckles, ulcers or flaps, and no change in the draining lymph node, whereas in Erysipelothrix- caused arthritis, the villous hypertrophy is severe and polypoid in nature, there is diffuse erosion of articular cartilage, and the draining lymph node is consistently hypertrophic and often cystic.     

The isolation of Streptococcus suis types 1 and 2 from pigs in New Zealand

Extract

Madam:– I wish to advise of the recent isolation of Streptococcus suis 1 and 2 from pigs in New Zealand. Type 1 was isolated from brain and joint swabs from a three-week-old piglet. The piglet had meningitis, was laterally recumbent, paddling and had arthritis with grossly swollen joints. Type 2 was isolated from tonsils collected from apparently normal bacon pigs slaughtered at a meat works. Isolates were confirmed as either Streptococcus suis type 1 or 2 on the basis of biochemical characteristics⁽⁴⁾ and Lancefield precipitin tests using Sand R antisera as originally used by De Moor.⁽²⁾     

Prevalence and age-related infection of Cryptosporidium suis, C. muris and Cryptosporidium pig genotype II in pigs on a farm complex in the Czech Republic

A total of 413 pig faecal samples were collected from pre-weaners (119), starters (131), pre-growers (123) and sows (40) from a farm with a closed breeding system segmented into two breeding complexes and a growing complex in the region of Vysočina, Czech Republic and screened for the presence of Cryptosporidium using staining methods and genotyping (SSU rRNA). Cryptosporidium oocysts were detected by microscopy in the faeces of 21.1% of the samples (87/413). Sequence analyses and RFLP identified C. suis in 44, Cryptosporidium pig genotype II in 23 and C. muris in 2 samples. No mixed infections were found.Pigs under 7 weeks of age were infected with C. suis only. Cryptosporidium pig genotype II was found in animals from 7 weeks of age. No relationship was found between diarrhoea and any Cryptosporidium infection in any of the different age groups (P < 0.05). The pre-weaned pigs shed significantly more Cryptosporidium oocysts than older pigs and it was associated with C. suis infection.     

Prevalence and Characteristics of Staphylococcus aureus in Connecticut Swine and Swine Farmers

This study explores the characteristics of Staphylococcus aureus (S. aureus) in swine and their human handlers in a convenience sample of 35 farms in Connecticut. Husbandry practices are clearly different from better‐known concentrated animal feeding operations (CAFOs) with less intensive rearing conditions. Nasal samples were collected from 263 pigs and nine humans on 35 farms during the 2010 rearing season. Samples were analysed using established microbiology methods, and resulting methicillin‐sensitive (MSSA) and resistant (MRSA) isolates were typed by pulsed‐field gel electrophoresis (PFGE) and spa typing. PCR was used to detect the presence of the Panton‐Valentine Leukocidin (PVL) gene, a cytotoxin usually associated with CA‐MRSA infection. A farm assessment form and questionnaire were used to obtain the information about husbandry practices and human exposure risk, respectively. Staphylococcus aureus colonized swine and humans were found in 51% (18/35) of the farms sampled at a rate of 30% (85/259) and 22% (2/9), respectively. Eight pigs and two humans were MRSA positive on five farms. MRSA in swine was related to healthcare‐associated (HA), community‐associated (CA) or livestock‐associated (LA) MRSA strains, whereas humans were colonized with HA‐MRSA. On the basis of spa typing, there was evidence of human–animal transmission thereby signifying humanosis/reverse zoonoses. The PVL gene was found in 88% (7/8) of MRSA swine isolates, the first time this gene has been seen in colonized pigs sampled on US farm. MSSA isolates belonged to six spa types: t337 (41%), t034 (12%), t334 (12%), t4529 (12%), t8760 (18%) and t1166 (6%) including LA strains. This is the first time spa type t8760 has been reported and the only MSSA with the PVL gene. In summary, MRSA including LA strains (LA‐MRSA) can be found on small farms with different husbandry practices from CAFOs, suggesting that preventive measures for zoonotic MRSA infection should address a range of animal production.