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Phage-encoded virulence genes of group A streptococci were detected in 10 (55.6%) of 18 isolates of group C streptococci that had caused bovine mastitis. Bovine isolates carried other genetic determinants, such as composite transposon Tn1207.3/F10394.4 (100%) and antimicrobial drug resistance genes erm(B)/erm(A) (22.2%), linB (16.6%), and tet(M)/tet(O) (66.7%), located on mobile elements. [ABSTRACT FROM AUTHOR]

The molecular epidemiology and population structure of 30 bovine subclinical mastitis field isolates of Streptococcus uberis, collected from 6 Portuguese herds (among 12 farms screened) during 2002 and 2003, were examined by using pulsed-field gel electrophoresis (PFGE) for clustering of the isolates and multilocus sequence typing (MLST) to assess the relationship between PFGE patterns and to identify genetic lineages. The 30 isolates were clustered into 18 PFGE types, using a similarity cutoff of 80%, and 3 PFGE types accounted for almost half of the isolates (46.6%). These major types were herd specific, suggesting either cow-to-cow transmission or infection with isolates from the same environmental reservoirs. The remaining unrelated PFGE types of isolates were from different herds strongly suggesting environmental sources of Strep. uberis infection. All 30 isolates were analyzed by MLST and clustered into 14 sequence types (ST). These ST were found to be novel, either with 10 new alleles of 6 housekeeping genes or with different combinations of previously assigned alleles. Five of these ST were clustered into 3 clonal complexes (lineages), ST-143, ST-86, and ST-5, known to includebovine isolates from several geographic locations (Australia, New Zealand, United Kingdom, Sweden, and Denmark) and 9 singletons. To our knowledge, this is the first report that documents molecular typing studies of bovine isolates of Strep. uberis from Portugal, which were shown to represent novel genomic backgrounds of this pathogen. [ABSTRACT FROM AUTHOR]

The records of 116 cattle suffering from cardiac disease were examined retrospectively. On the basis of the results of postmortem examinations there were 52 cases of endocarditis, 39 of pericarditis and 25 congenital cardiac defects. The most useful clinical tool for differentiating between these conditions was auscultation of the heart. The cases of pericarditis were characterised by muffled heart sounds, and the cases of endocarditis and congenital cardiac defects were characterised by a cardiac murmur. Endocarditis could be differentiated from congenital cardiac defects by the presence of a jugular pulse, venous distension, oedema, a reduced appetite, pain and polyarthritis, whereas congenital defects were associated with conformational abnormalities. These two conditions could also be differentiated by differences in the plasma sodium concentration, the albumin:globulin ratio, red blood cell count, lymphocyte count and haematocrit. The ability to differentiate between these three groups of cardiac diseases can help the veterinary practitioner in deciding whether treatment, economic salvage (slaughter for human consumption) or disposal (slaughter not for human consumption) is likely to be the best option. [ABSTRACT FROM AUTHOR]

To evaluate the antimicrobial resistance traits of staphylococci responsible for subclinical bovine mastitis in Portugal, the minimum inhibitory concentrations (MIC) of 7 antimicrobial agents, frequently administered for mastitis treatment, were determined for 30 Staphylococcus aureus and 31 Staphylococcus epidermidis field isolates, β-Lactamase production was detected through the use of nitrocefin-impregnated discs. The MIC that inhibited 90% of the isolates tested (MIC90) of penicillin, oxacillin, cefazolin, gentamicin, sulfamethoxazole/trimethoprim, oxytetracycline, and enrofloxacin were, respectively, 4, 0.5, 1, 1, 0.25, 0.25, and 0.06 µg/mL for Staph. aureus and ≥64, 8, 1, 32, ≥64, ≥64, and 0.06 µg/mL for Staph. epidermidis. All Staph. aureus isolates showed susceptibility to oxacillin, cefazolin, gentamicin, sulphamethoxazole/trimethoprim, and enrofloxacin, β-Lactamase production was detected in 20 of these isolates (66.7%), all of which were resistant to penicillin. Of the 31 Staph. epidermidis tested, 24 (77.4%) were β-lactamase positive. All isolates were susceptible to both cefazolin and enrofloxacin. Nine Staph. epidermidis isolates were resistant to oxacillin, with MIC values ranging from 4 to 8 µg/mL. The MIC values of 5 antimicrobial agents tested were higher than those reported in other countries. Enrofloxacin was the only exception, showing lower MIC values compared with other reports. Overall, the antimicrobial agents tested in our study, with the exception of penicillin, were active against the 61 isolates studied. [ABSTRACT FROM AUTHOR]

Abstract: Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa=0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa=0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production. [Copyright &y& Elsevier]

The article describes the clinical findings typical of cases of malignant catarrhal fever (MCF) and mucosal disease, and compares these with clinical findings characteristic of the bluetongue outbreak in northern Europe. The clinical signs for which there was a significant difference in the proportion of cases presenting with each disease were appetite, demeanour, size for age, fecal appearance, adventitious lung sounds, corneal opacity, nasal discharge, halitosis, and interdigital ulceration.

Methicillin-resistant staphylococci (MRS) have already been reported as mastitis agents. Such bacterial species are a public health concern, and the characterization of their antimicrobial resistance and virulence profile is important to better control their dissemination. The present work evaluated the distribution of methicillin-resistance among 204 staphylococci from clinical (n = 50) and subclinical (n = 154) bovine mastitis. The presence of the mecA gene was determined by PCR. Phenotypic expression of coagulase, DNase, lipase, gelatinase, hemolytic enzymes, and biofilm production was evaluated. The presence of biofilm-related genes, icaA, icaD, and bap, was also determined. Antimicrobial resistance patterns for aminoglycosides, lincosamides, macrolides, fluoroquinolones, sulphonamides, tetracyclines, and fusidic acid were determined. Nineteen (9.3%) isolates were identified as MRS, and the presence of mecA in these isolates was confirmed by PCR. Virulence factors evaluation revealed that gelatinase was the most frequently detected (94.7%), followed by hemolysins (73.7%) and lipase (68.4%); 84.2% of the MRS isolates produced biofilm and icaA and icaD were detected in almost half of the MRS isolates (52.6%), but all were bap-negative. Resistance against other antimicrobial agents ranged from 0 (fusidic acid, ciprofloxacin, norfloxacin, enrofloxacin) to 100% (nalidixic acid). Resistance to nalidixic acid and nalidixic acid-tetracycline were the most common antimicrobial resistance profiles (31.6%). This study confirms that despite the low prevalence of MRS, isolates frequently express other virulence traits, especially biofilm, that may represent a serious challenge to clinicians. [ABSTRACT FROM AUTHOR]

A 12-month study was conducted to assess and monitor gastrointestinal tract nematodes and liver fluke in cohorts of cattle on a Scottish organic dairy farm. Various diagnostic markers for helminth parasites of cattle from different age groups were assessed monthly from April 2007 to March 2008. First season grazing stock were subjected to significant challenge from Ostertagia ostertagi nematodes as reflected in serum pepsinogen concentrations, which rose markedly in the second half of the grazing season. In addition, plasma albumin concentrations decreased and faecal egg counts (FEC) increased moderately, indicating exposure to both O ostertagi and probably Cooperia oncophora. Second season grazing animals had a peak FEC early in the grazing period, suggestive of a potential carry-over of Ostertagia species infection ('Type 2') during housing. All classes of cattle showed evidence of fluke (Fasciola hepatica) infection. Adult cow exposure to O ostertagi and fluke was estimated via the use of ELISA testing to detect antibodies to O ostertagi and F hepatica and the high levels detected suggested a significant exposure response. Despite low stocking densities and sympathetic grazing management, there was a significant challenge to all grazing stock from gastrointestinal nematodes and liver fluke. [ABSTRACT FROM AUTHOR]

Renal dysplasia and nephrosclerosis in six calves, which were aged three to six months and from different farms in western Scotland and north-west England, was characterised clinically by stunted growth and renal failure with uraemia. Affected animals were depressed and one case exhibited severe neurological signs. Reduced erythrocyte counts were evident in three of four animals from which blood samples were submitted for haematology. At postmortem examination, the kidneys were bilaterally small, pale and firm, with marked fibrosis and sometimes contraction of the capsule. Histologically, affected calves had disorganised atrophic glomeruli, dilatation of tubules, loss of nephrons, areas of undifferentiated mesenchyme and diffuse interstitial and periglomerular fibrosis. There was minimal inflammation. Renal dysplasia and nephrosclerosis is a form of juvenile nephropathy of unknown aetiology that occurs sporadically in calves in the UK. [ABSTRACT FROM AUTHOR]

Background: Making a clinical diagnosis of pericarditis in cattle is difficult and additional diagnostic tests are needed to evaluate cattle with suspected pericarditis. Serum cardiac troponin I (cTnI) concentrations are increased in cattle with pericarditis, but the utility of measuring serum cTnI concentrations in cattle with suspected pericarditis in cattle remains unclear. Objectives: To determine if serum cTnI concentrations in cattle can be used to differentiate pericarditis from other cardiac disorders and noncardiac thoracic diseases. Animals: Seventy-seven clinically diseased cattle and 19 healthy control cattle. Methods: Serum cTnI concentrations were measured using an Immunlite Troponin I immunometric chemiluminescent assay in consecutive cases of postmortem-confirmed pericarditis (n = 18), endocarditis (n = 15), chronic suppurative pneumonia (n = 13), congenital heart disease (n = 10), reticulitis (n = 3), mediastinal abscess (n = 7), thymic lymphoma (n = 6), and caudal vena cava thrombosis (n = 5). Serum cTnI concentrations were measured in 19 healthy cattle. Results: Although serum cTnI concentrations were significantly higher in cattle with pericarditis compared with healthy cattle, they were not significantly different from concentrations in cattle with endocarditis, congenital cardiac disease, mediastinal abscess, reticulitis, caudal vena cava thrombosis, or chronic suppurative pneumonia. Conclusions: Serum cTnI cannot be used to distinguish cattle with pericarditis from cattle with other primary cardiac diseases. In addition, serum cTnI concentrations cannot distinguish between cattle with primary cardiac diseases and those with other noncardiac, intrathoracic disorders. [ABSTRACT FROM AUTHOR]

Intramammary infection (IMI), also known as mastitis, is the most frequently occurring and economically the most important infectious disease in dairy cattle. This study provides a validation of the analytical specificity and sensitivity of a real-time PCR-based assay that identifies 11 major pathogen species or species groups responsible for IMI, and a gene coding for staphylococcal β-lactamase production (penicillin resistance). Altogether, 643 culture isolates originating from clinical bovine mastitis, human, and companion animal samples were analyzed using the assay. The isolates represented 83 different species, groups, or families, and originated from 6 countries in Europe and North America. The analytical specificity and sensitivity of the assay was 100% in bacterial and β-lactamase identification across all isolates originating from bovine mastitis (n = 454). When considering the entire culture collection (including also the isolates originating from human and companion animal samples), 4 Streptococcus pyogenes, 1 Streptococcus salivarius, and 1 Streptococcus sanguis strain of human origin were identified as Streptococcus uberis, and 3 Shigella spp. strains were identified as Escherichia coli, decreasing specificity to 99% in Strep. uberis and to 99.5% in E. coli. These false-positive results were confirmed by sequencing of the 16S rRNA gene. Specificity and sensitivity remained at 100% for all other bacterial targets across the entire culture collection. In conclusion, the real-time PCR assay shows excellent analytical accuracy and holds much promise for use in routine bovine IMI testing programs. This study provides the basis for evaluating the assay's diagnostic performance against the conventional bacterial culture method in clinical field trials using mastitis milk samples. [ABSTRACT FROM AUTHOR]

Abstract: Biofilm formation is considered a selective advantage for staphylococci mastitis isolates, facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers and enclosing in a polymeric matrix, being regulated by several loci. As biofilm formation can proceed through different pathways and time ranges, its detection may differ according to the time of observation. This study aimed at evaluating the time course evolution of biofilm production in Staphylococcus aureus (n =26) and Staphylococcus epidermidis (n =29) mastitis isolates by Fluorescent In Situ Hybridisation. Biofilm-forming ability increased with incubation time for both species: for S. aureus, 34.6%, 69.2% and 80.8% of the isolates were able to produce biofilm at 24, 48 and 72h, respectively. For S. epidermidis, 44.8%, 62.1% and 75.9% of the isolates were biofilm-positive at 24, 48 and 72h, respectively. No significant difference was found between species at each time point (Friedman''s test, p >0.05). For S. aureus, although a significant difference was found between 24 and 48h (Wilcoxon matched paired test, p <0.05), no significant difference was found between 24 and 48h (p >0.05). For S. epidermidis, significant differences were found between each time point (p <0.05). Bacterial biofilms may impair eradication of chronic mastitis, rendering antibiotherapy less effective. Detection of biofilm-forming ability in mastitis isolates may provide useful information for the establishment of a more adequate therapeutic regimen, in view of the antimicrobial concentrations required for bacterial control. However, it is essential that biofilm formation time course is taken into consideration. [Copyright &y& Elsevier]

The article provides information on a Salmonella enterica serovar Dublin (S. Dublin) outbreak in a dairy farm in Glasgow, Scotland in August 2006. A 70-year-old male developed salmonellosis due to close contact with livestock and drinking unpasteurized milk form the bulk milk tank on a daily basis prior to the outbreak. The need to consider S. Dublin among the differentials during abortion investigations is emphasized. One suggested way to prevent the risk of exposure of animals to S. Dublin is improving calf management.