In vitro identification and verification of inflammatory biomarkers in swine

Currently there are no non-steroidal anti-inflammatory drugs (NSAIDs) approved for the control of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess drug efficacy. This study investigates the differential expression of genes altered in response to Escherichia coli lipopolysaccharide (LPS) induced inflammation which may serve as indicators of NSAID efficacy. Unstimulated whole blood from swine was mixed with tissue culture media, stimulated with LPS, and RNA extracted at the following time points 0 h, 1 h, 3 h, 24 h and 48 h. Total RNA was extracted and analyzed using a commercial swine DNA microarray. The DNA microarray was utilized as a screen to determine potential biomarkers, focusing on the genes that exhibited the greatest degree of differential expression. A master list of 57 genes was formed based on the differential expression as a result of the stimulation. Following analysis, 12 genes whose expressions were significantly altered (8 up- and 4 down-regulated) were chosen for verification via quantitative RT-PCR (qRT-PCR). The qRT-PCR analysis confirmed the differential expression of 11 of the 12 genes chosen via the microarray analyses. Specifically, traditional genes such as SAA, G-CSF, and IL-10 were up-regulated, while CD4 was down-regulated; all of the genes were altered by 24 h or 48 h post-stimulation. We demonstrate here that expression of these 11 genes is altered as a direct result of LPS stimulation and consequently inflammation.     

Antimicrobial resistant Escherichia coli isolates in cattle and house sparrows on two Czech dairy farms

Rectal smears of calves, cows and young bulls, as well as cloacal smears of house sparrows (Passer domesticus), from farms at the villages of Šumice and Troskotovice, Czech Republic, were examined for E. coli resistant to 12 antimicrobials. The resistant isolates were tested for antimicrobial-resistance genes and integrons. Totals of 40% (n = 183), 3% (n = 95), 0% (n = 33), and 9% (n = 54) of Escherichia coli isolates from calves, cows, young bulls and house sparrows, respectively, were antimicrobial resistant. The following genes were identified in cattle E. coli isolates: tetA, tetB (isolates resistant to tetracycline), bla_TEM (beta-lactams), strA, aadA (streptomycin), sul1, sul2 (sulphonamides), and cat, floR (chloramphenicol). Seven of 16 antimicrobial-resistant calf isolates from the Šumice farm possessed class 1 integrons with the aadA1 gene cassette integrated, 1 kb in size. On the Troskotovice farm, eight of 57 antimicrobial-resistant calf isolates possessed class 1 integrons. Integrons of 1.5 kb with the dhfr1- aadA1 gene cassette were found in four isolates, followed by a 1 kb integron with the aadA1 gene found in three isolates, and a 1.7 kb integron with the dhfr17-aadA5 gene cassette and the phenotype ASSuTSxtNaCipCCfG. The prevalence of resistant E. coli in calves compared to adult cattle was much higher and probably was influenced by oral antimicrobial usage in calves, feeding with milk and colostrum from treated cows, as well as mechanisms unrelated to antimicrobial drug selection. Although house sparrows lived together with the cattle and came into contact with cattle waste on the farm, they were not infected by resistant E. coli isolates with the same characteristics as those found in cattle.     

Effect of training status on immune defence related gene expression in Thoroughbred: Are genes ready for the sprint?

Athletic performance is both a stress factor and an adaptive response to exercise that may be modulated by training, reduce inflammation and help prevent disease. Studies on the endocrinology of exercise and training have demonstrated the existence of an integrated metabolic network of hormone and cytokine regulation. Subsequent molecular studies have shown that repeated bouts of exercise may establish new basal levels of gene expression at rest.The Thoroughbred horse may be a useful ‘exercise model’ for inter-individual comparisons between subjects with homogeneous genetic and environmental backgrounds and similar exercise management practices. In this study, the effects of training and acute effort on gene expression were evaluated with a real time PCR approach in athletic (n = 10) and sedentary horses (n = 9), using a previously characterised panel of genes known to be highly modulated during effort (CXCL2, TLR4, IL1β, IL8, IL1RII, IL18, IL6 and CEBPβ). A ‘rest comparison’ was performed to evaluate a training effect in both groups while a ‘race comparison’ was performed in athletic horses only (before, immediately after, and 12 h after racing) to determine the effect of acute effort.The results indicated that many of the investigated genes (TLR4, IL1β, IL1RII, IL18, IL6 and CEBPβ) were expressed to a greater extent in athletic horses compared to sedentary animals when both were at rest. However, a time-course comparison in the athletic horses revealed that genes exhibiting the highest levels of expression at rest did not show significant changes after the race. The findings suggested that training may exert a conditioning on gene expression at rest leading to a more prompt response to exercise-induced stress in Thoroughbreds.     

Antimicrobial resistance changes in enteric Escherichia coli of horses during hospitalisation: Resistance profiling of isolates

The aim of this study was to determine whether hospitalisation of horses leads to increased antimicrobial resistance in equine faecal Escherichia coli isolates. E. coli were cultured from faecal samples of horses on admission and after 7 days of hospitalisation; antimicrobial susceptibility was determined for eight antimicrobial agents. Resistance profiles of E. coli isolates were grouped into clusters, which were analysed to determine resistance patterns. Resistance to 7/8 antimicrobial agents and multi-drug resistance (MDR; resistance to ?3 antimicrobial classes) were significantly higher after 7 days of hospitalisation. Forty-eight resistance profiles were identified; 15/48 were present on day 0 only, 16/48 on day 7 only and 17/48 at both times of sampling. There was a significant association between day 7 profiles and resistance detected to an increased number of antimicrobial agents. Hospitalisation of horses for 7 days resulted in alterations in equine faecal E. coli antimicrobial resistance profiles.     

Molecular characterization of quinolone resistance mechanisms and extended-spectrum β-lactamase production in Escherichia coli isolated from dogs

The increasing prevalence of antimicrobial resistances is now a worldwide problem. Investigating the mechanisms by which pets harboring resistant strains may receive and/or transfer resistance determinants is essential to better understanding how owners and pets can interact safely. Here, we characterized the genetic determinants conferring resistance to β-lactams and quinolones in 38 multidrug-resistant Escherichia coli isolated from fecal samples of dogs, through PCR and sequencing. The most frequent genotype included the β-lactamase groups TEM (n = 5), and both TEM + CTX-M-1 (n = 5). Within the CTX-M group, we identified the genes CTX-M-32, CTX-M-1, CTX-M-15, CTX-M-55/79, CTX-M-14 and CTX-M-2/44. Thirty isolates resistant to ciprofloxacin presented two mutations in the gyrA gene and one or two mutations in the parC gene. A mutation in gyrA (reported here for the first time), due to a transversion and transition (TCG → GTG) originating a substitution of a serine by a valine in position 83 was also detected. The plasmid-encoded quinolone resistance gene, qnrs1, was detected in three isolates. Dogs can be a reservoir of genetic determinants conferring antimicrobial resistance and thus may play an important role in the spread of antimicrobial resistance to humans and other co-habitant animals.     

Higher resistance of Campylobacter coli compared to Campylobacter jejuni at chicken slaughterhouse

In order to compare the prevalence of Campylobacter coli and Campylobacter jejuni during the processing of broilers at slaughterhouse a total of 848 samples were analyzed during 2012 in southern Spain. Four hundred and seventy six samples were collected from cloaca, carcass surfaces and quartered carcasses. Moreover, 372 environmental swabs from equipment and scalding water were collected. Minimum inhibitory concentration (MIC) to ciprofloxacin, erythromycin, streptomycin, tetracycline and gentamicin was determined for isolates from chicken meat. The general prevalence of Campylobacter was 68.8% (40.2% of C. coli and 28.5% of C. jejuni). The relative prevalence of C. coli increased from loading dock area (41.5%) to packing area (64.6%). In contrast, the relative prevalence of C. jejuni decreased from 58.5% to 35.4%. These differences between species from initial to final area were significant (p = 0.02). The highest antimicrobial resistance for C. jejuni and C. coli was detected to tetracycline (100%) and ciprofloxacin (100%), respectively. Campylobacter coli showed an antimicrobial resistance significantly higher than C. jejuni to streptomycin (p = 0.002) and erythromycin (p < 0.0001).     

Frequency of virulence factors in Escherichia coli isolated from suckling pigs with diarrhoea in China

Escherichia coli-associated diarrhoea is an important disease adversely affecting the pig industry. This study was conducted to investigate the frequency of virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China. A total of 381 E. coli strains, obtained from 290 faecal samples from pigs on 38 farms, were tested for fimbriae (K88, K99, 987P, F41, F18, F17), non-fimbrial adhesins (AIDA-I, paa, CS31A, eae, saa), enterotoxin (LT-I, LT-II, STa, STb, EAST1), Shiga toxin (Stx1, Stx2, Stx2e), pathogenicity islands (HPI, LEE), α-haemolysin (hlyA), afa8 gene cluster (afaD, afaE) and sepA genes by PCR. Out of the 381 isolates, 206 carried at least one virulence gene. Of the 206 virulence positive isolates, the virulence factor genes detected were EAST1 (n = 120), irp2 (n = 59), paa (n = 50), STb (n = 41), AIDA-I (n = 34), LT-I (n = 23), ler (n = 11), hlyA (n = 9), K88 (n = 8), eae (n = 8), STa (n = 7), sepA (n = 6), F18 (n = 5), afaD (n = 3), afaE (n = 3), K99 (n = 2) and Stx2e (n = 1), with most isolates carrying multiple virulence genes. These results demonstrate that relatively few isolates from the study population express K88, K99, LT-I or STa, but that EAST1 (58%), irp2 (29%), AIDA-I (16.5%), paa (24%) and STb (20%) are frequent virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China.     

Serum amyloid A measurements in saliva and serum in growing pigs affected by porcine respiratory and reproductive syndrome in field conditions

The response of serum amyloid A (SAA) concentrations in serum and saliva during a natural porcine respiratory and reproductive syndrome (PRRS) outbreak was investigated. Basal levels of SAA were reliably determined and significant differences in SAA were found between diseased and healthy pigs in both sample types (serum SAA medians 78.3 vs. 55.74 mg/L, respectively; salivary SAA medians 8.97 vs. 2.52 mg/L, respectively; P < 0.001). In serum, an SAA cut-off value of 66.4 mg/L showed 68.9% sensitivity and 68.09% specificity, while in saliva an SAA cut-off value of 5.59 mg/mL showed a sensitivity–specificity pair of 69–66%. Furthermore, it was observed that the growth stage of animals should be accounted to correctly interpret SAA measurements, since more accurate cut-off values could be determined and a particular behaviour of salivary SAA was identified in post-weaning pigs. Salivary and serum SAA measurements can therefore be confirmed as a valuable tool to consistently discriminate between healthy and PRRS-affected pigs.     

Giardia and Cryptosporidium infections in neonatal reindeer calves: Relation to the acute phase response

This longitudinal observational study was conducted to investigate the spontaneous effect of Giardia and Cryptosporidium infections on acute phase response (APR) in reindeer calves (Rangifer tarandus tarandus) in Finnish Lapland.Serum (n = 609) and faecal samples (n = 366) were collected from 54 reindeer calves aged zero to 33 days. The samples were analysed for Giardia, Cryptosporidium, acute phase proteins (APP) and γ-globulins.Linear regression models were used to investigate associations of early Giardia infection (before 12 days of life) with the response of APPs and acquiring of passive immunity.Giardia was detected in 100% and Cryptosporidium in 23% of calves. There was a negative association between early Giardia infection and γ-globulin concentrations (p = 0.032) and a positive association with serum amyloid A (SAA) concentrations (p = 0.042). The results suggest a protective effect of colostrum against Giardia infection and that early infection may induce activation of APR.     

Lactobacillus reuteri ATCC 55730 and L22 display probiotic potential in vitro and protect against Salmonella-induced pullorum disease in a chick model of infection

Lactobacillus reuteri ATCC 55730 (L. reuteri ATCC 55730) and L. reuteri L22 were studied for their probiotic potential. These two strains were able to produce an antimicrobial substance, termed reuterin, the maximum production of reuterin by these two strains was detected in the late logarithmic growth phase (16 h in MRS and 20 h in LB broths). These two strains could significantly reduce the growth of Salmonella pullorum ATCC 9120 in MRS broth, L. reuteri ATCC 55730 with a reduction of 48.2 ± 4.15% (in 5 log) and 89.7 ± 2.59% (in 4 log) respectively, at the same time, L. reuteri L22 was 69.4 ± 3.48% (in 5 log) and 80.4 ± 3.22% respectively. L. reuteri ATCC 55730 was active against the majority of the pathogenic species, including S. pullorum ATCC 9120 and Escherichia coli O₇₈, while L. reuteri L22 was not as effective as L. reuteri ATCC 55730. The two potential strains were found to survive variably at pH 2.5 and were unaffected by bile salts, while neither of the strains was haemolytic. Moreover, L. reuteri ATCC 55730 exhibited variable susceptibility towards commonly used antibiotics; but L. reuteri L22 showed resistant to most antibiotics in this study. L. reuteri ATCC 55730 consequently was found to significantly increase survival rate in a Salmonella-induced pullorum disease model in chick. To conclude, strain L. reuteri ATCC 55730 possesses desirable probiotic properties, such as antimicrobial activity and immunomodulation in vitro, which were confirmed in vivo by the use of animal models.     

Resveratrol decreases oxidative burst capacity and alters stimulated leukocyte cytokine production in vitro

IntroductionResveratrol, a naturally-occurring phytophenol, has been shown to bolster immune surveillance and reverse immunosenescence in a dose dependent manner in rodents and humans. Although safety and pharmacokinetic studies have been completed in dogs, the immunomodulatory effects of resveratrol in dogs has not yet been investigated. The objective of this study was to determine the effect of resveratrol on canine innate immune system function in vitro. The hypothesis was that similar to other species, low concentrations of resveratrol would stimulate while high concentrations would depress innate immune system function.MethodsWhole blood was collected from six healthy, adult, client-owned dogs and was incubated with resveratrol at final concentrations of 6000 ng ml⁻¹, 3000 ng ml⁻¹, 1000 ng ml⁻¹, or control solution for 4 h. Following incubation, phagocytosis and oxidative burst were evaluated using flow cytometry, and LPS-, lipoteichoic acid (LTA) – and peptidoglycan (PG)-stimulated leukocyte production of TNF, IL-6, and IL-10 were measured using a canine specific multiplex assay.ResultsPhagocytosis was not altered by resveratrol at any concentration compared to control. However, while the number of PMNs capable of performing oxidative burst did not change, the robustness of the reaction following stimulation with Escherichia coli and PMA was reduced in a dose dependent manner. In addition, LPS-, LTA-, PG, and PBS-stimulated TNF production was increased following incubation with all concentrations of resveratrol compared to control, and this effect was dose dependent. LTA-stimulated IL-6 was increased with resveratrol compared to control. Furthermore, LTA-stimulated IL-10 was decreased with 6000 ng ml⁻¹ and 3000 ng ml⁻¹ concentrations of resveratrol and PG-stimulated IL-10 production was decreased with all concentrations of resveratrol compared to control. The LPS-, LTA-, and PG-stimulated TNF:IL-10 ratio was increased with 6000 ng ml⁻¹ of resveratrol compared to control and lower resveratrol concentrations.ConclusionWhile resveratrol was sparing to PMN phagocytosis, it reduced the robustness of PMN oxidative burst. Resveratrol also increased pro-inflammatory and decreased anti-inflammatory leukocyte cytokine production capacity in vitro. These data suggest that resveratrol supplementation may depress oxidative burst reactions while promoting pro-inflammatory leukocyte cytokine production and decreasing anti-inflammatory cytokine production. Based on these findings, further in vivo study regarding the effects of resveratrol on PMN oxidative burst capability and leukocyte cytokine production capacity are indicated prior to routine supplementation.     

Characterization of colonization-deficient mutants of Actinobacillus suis

Actinobacillus suis is an important opportunistic pathogen of swine that can cause disease in pigs of all ages, especially in high-health status herds. Although A. suis shares many virulence factors in common with Actinobacillus pleuropneumoniae and can cause a haemorrhagic pleuropneumonia similar to that caused by A. pleuropneumoniae, A. suis most often causes septicaemia and diseases such as arthritis and meningitis that are sequelae to septicaemia. In a recent signature-tagged transposon mutagenesis study, 30 colonization-essential genes of A. suis were identified. In the current study, the attachment and invasion patterns of strains harboring Tn10 insertions in ompA, pfhaB1, lcbB, and cpxR were evaluated using porcine palatine tonsil organ cultures, the swine kidney epithelial cell line, SK6, and a porcine brain microvascular endothelial cell line, PBMEC/C1-2. All of these mutants attached in lower numbers than wild type to the tonsillar explants and to the SK6 cells. The ompA mutant attached in significantly lower numbers than wild type to the porcine tonsil cells (P = 0.02) and to PBMEC (P = 0.0008) at 60 min time point. As well, the ompA mutant showed significantly greater sensitivity than wild type to chemical stressors and to swine serum. Using fluorescent microscopy, a GST-OmpA fusion protein could be demonstrated to interact with the crypt epithelial cells of porcine palatine tonsil.     

Characterisation of ceftiofur resistance in swine bacterial pathogens

A PCR based method was developed for the identification of ceftiofur resistance genes (bla_CMY-2, bla_TEM-1, and ampC) in swine bacterial pathogens. Using this method, the ceftiofur resistant (n = 76) and susceptible (n = 45) strains of Bordetella bronchiseptica, Salmonella spp., Escherichia coli, and Pasteurella multocida were screened for the presence of these three genes. The resistant genes were detected in 70% (bla_TEM-1), 68% (bla_CMY-2) and 45% (ampC) of the resistant isolates and in 18% (bla_TEM-1), 27% (bla_CMY-2), and 36% (ampC) of the susceptible isolates. Results obtained in the present study showed widespread distribution of these three resistance genes in ceftiofur-resistant swine pathogens. It was also observed that more pathogens are acquiring these resistance genes.     

Comparison of paired serum and lithium heparin plasma samples for the measurement of serum amyloid A in horses using an automated turbidimetric immunoassay

This study aimed to evaluate whether equine serum amyloid A (SAA) concentrations could be reliably measured in plasma with a turbidimetric immunoassay previously validated for equine SAA concentrations in serum. Paired serum and lithium-heparin samples obtained from 40 horses were evaluated. No difference was found in SAA concentrations between serum and plasma using a paired t test (P = 0.48). The correlation between paired samples was 0.97 (Spearman’s rank P < 0.0001; 95% confidence interval 0.95–0.99). Passing-Bablok regression analyses revealed no differences between paired samples. Bland–Altman plots revealed a positive bias in plasma compared to serum but the difference was not considered clinically significant. The results indicate that lithium-heparin plasma samples are suitable for measurement of equine SAA using this method. Use of either serum or plasma allows for greater flexibility when it comes to sample collection although care should be taken when comparing data between measurements from different sample types.     

Bovine natural killer cells are present in Escherichia coli infected mammary gland tissue and show antimicrobial activity in vitro

Natural killer (NK) cells are early responders in bacterial infections but their role in bovine mastitis has not been characterized. For the first time, we show the presence of NK cells (NKp46⁺/CD3⁻) in bovine mammary gland tissue after an intramammary challenge with Escherichia (E.) coli. A small number of NK cells was detected in milk from quarters before and during an E. coli challenge. In vitro cultures of primary bovine mammary gland epithelial cells stimulated with UV irradiated E. coli induced significant migration of peripheral blood NK cells (pbNK) within 2 h. Furthermore, pbNK cells significantly reduced counts of live E. coli in vitro within 2 h of culture. The results show that bovine NK cells have the capacity to migrate to the site of infection and produce antibacterial mediators. These findings introduce NK cells as a leukocyte population in the mammary gland with potential functions in the innate immune response in bovine mastitis.     

The use of eugenol against Aeromonas hydrophila and its effect on hematological and immunological parameters in silver catfish (Rhamdia quelen)

The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L^?1), it promoted survival in infected silver catfish. Eugenol (50 μg mL^?1) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L^?1) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish.     

Expression of trefoil factor genes in the duodenum and colon of dogs with inflammatory bowel disease and healthy dogs

Trefoil factors (TFF) are small peptides produced by goblet cells, which are crucial for epithelial restitution. In humans with inflammatory bowel disease (IBD), TFF expression is up-regulated as part of an unspecific repair mechanism. The goal of this study was to assess TFF gene expression in the gastrointestinal tract from dogs with IBD compared to healthy controls. Preliminary assessment by PCR revealed TFF1 and 3 expression in the small and large intestine, whereas TFF2 was amplified only in the stomach. Subsequent RT-qPCR (with relative quantification against 3 reference genes) on endoscopic duodenal (IBD n = 22, healthy controls n = 18) and colonic (IBD n = 12, controls n = 11) biopsies revealed that TFF1 expression was significantly up-regulated in the duodenum from IBD dogs (Mann–Whitney p = 0.001), whereas TFF3 expression was significantly lower in IBD colon compared to controls (t-test p = 0.018).This study demonstrates evidence for dysregulation of TFF gene expression in canine IBD. Up-regulation of TFF1 could signify ectopic expression as a compensatory repair-mechanism, whereas down-regulation of TFF3 could contribute to defective epithelial barrier function, respectively. Whether this is a cause or consequence of IBD could not be established.     

Inverse relationship between heat stable enterotoxin-b induced fluid accumulation and adherence of F4ac-positive enterotoxigenic Escherichia coli in ligated jejunal loops of F4ab/ac fimbria receptor-positive swine

Heat-labile enterotoxin (LT) produced by enterotoxigenic Escherichia coli (ETEC) increases bacterial adherence to porcine enterocytes in vitro and enhances small intestinal colonization in swine. Heat-stable enterotoxin-b (STb) is not known to affect colonization; however, through an induction of net fluid accumulation it might reduce bacterial adherence. The relationship between fluid accumulation and bacterial adherence in jejunal loops inoculated with ETEC strains that produce LT, STb, both, or neither toxin was studied. Ligated jejunal loops were constructed in weaned Yorkshire pigs in two independent experiments (Exp. 1, n = 5, 8-week-old; Exp. 2, n = 6, 6–8-week-old). Each pig was inoculated with six F4ac⁺ E. coli strains: (1) LT⁺, STb⁺ parent (WAM2317); (2) STb^? (ΔestB) mutant (MUN297); (3) MUN297 complemented with STb (MUN298); (4) LT^? STb^? (ΔeltAB ΔestB) mutant (MUN300); (5) MUN300 complemented with LT (MUN301); and (6) 1836-2 (non-enterotoxigenic, wild-type). Pigs were confirmed to be K88 (F4)ab/ac receptor-positive in Exp. 2 by testing for intestinal mucin-type glycoproteins and inferred to be receptor-positive in both Exp. 1 and 2 based on histopathologic evidence of bacterial adherence. Strains that produced STb induced marked fluid accumulation with the response (ml/cm) to WAM2317 and MUN298 significantly greater than that to the other strains (P < 0.0001). Conversely, bacterial adherence scores based on immunohistochemistry and CFU/g of washed mucosa were both lowest in the strains that expressed STb and highest in those that did not. For the two experiments combined, the Pearson correlation coefficient (R) between fluid volume (ml/cm) and log CFU per gram was ?0.57021 (P < 0.0001); R² = 0.3521 (n = 197). These results support the hypothesis that enterotoxin-induced fluid accumulation flushes progeny organisms into the lumen of the bowel, thereby increasing the likelihood of fecal shedding and transmission of the pathogen to new hosts.     

Detection of classical and newly described staphylococcal superantigen genes in coagulase-negative staphylococci isolated from bovine intramammary infections

The coagulase-negative staphylococci (CNS) are the most prevalent mastitis pathogen group yet their virulence characteristics have not been well described. We investigated the presence of 19 classical and newly described staphylococcal superantigen (SAg) genes in CNS isolates from bovine intramammary infections (IMI). A total of 263 CNS representing 11 different Staphylococcus spp. were examined, and 31.2% (n = 82) of CNS isolates had one or more SAg genes; there were 21 different SAg gene combinations. The most prevalent combination of SAg genes (seb, seln and selq; n = 45) was found in S. chromogenes, S. xylosus, S. haemolyticus, S. sciuri subsp. carnaticus, S. simulans and S. succinus. The genes for SAgs appear to be widely distributed amongst CNS isolated from bovine IMI.