Antimicrobial resistance and molecular epidemiology of streptococci from bovine mastitis

Streptococcus agalactiae (Group B Streptococcus, GBS), Streptococcus dysgalactiae subsp. dysgalactiae (Group C Streptococcus, GCS) and Streptococcus uberis are relevant mastitis pathogens, a highly prevalent and costly disease in dairy industry due to antibiotherapy and loss in milk production. The aims of this study were the evaluation of antimicrobial drug resistance patterns, particularly important for streptococcal mastitis control and the identification of strain molecular features. Antimicrobial resistance was assessed by disk diffusion against amoxicillin–clavulanic acid, cefazolin, cefoperazone, pirlimycin-PRL, rifaximin, streptomycin, chloramphenicol, erythromycin-ERY, gentamicin, tetracycline-TET and vancomycin. Genotypic relationships were identified using pulsed-field gel electrophoresis (PFGE), macrolide and/or tetracycline resistance gene profiling, GBS capsular typing, GBS virulence gene profiling and GBS and S. uberis multi locus sequence typing (MLST).The majority of the isolates were susceptible to all drugs except to aminoglycoside, macrolide, lincosamide and tetracycline. Close to half of the TET resistant isolates have tetO and tetK and almost all ERY–PRL resistant isolates have ermB. A high degree of intra-species polymorphism was found for GCS. The GBS belonged to ST-2, -554, -61, -23 lineages and five new molecular serotypes and human GBS insertion sequences in the cpsE gene were found. Also, GBS of serotype V with scpB and lmb seem to be related with GBS isolates of human origin (same ST-2 and similar PFGE). Overall our results suggested that different therapeutic programs may have been implemented in the different farms and that in most cases clones were herd-specific.     

Teleost fish osteocalcin 1 and 2 share the ability to bind the calcium mineral phase

The occurrence of a second osteocalcin (OC2) has been reported in teleost fish, where it coexists with OC1 in some species. While it has been proposed that OC2 gene originated from OC1 through the fish whole-genome duplication event, little information is available on its molecular function and physiological role. The present study brings biological data supporting the presence of OC2 in the mineral phase of teleost fish bone and its association with the mineral phase together with OC1. The occurrence of OC2 forms with different levels of phosphorylation or γ-carboxylation, and with amino acid substitutions was observed. Comparative analysis of mature peptide sequences revealed the high conservation existing between OC1 and OC2, in particular within the core γ-carboxyglutamic acid domain, and suggests that both protein forms may have the same function, i.e., binding of calcium ions or hydroxyapatite crystals.     

Histology,immunocytochemistry and qRT‐PCR analysis of Atlantic salmon,Salmo salar L., post‐smolts following infection with infectious pancreatic necrosis virus (IPNV)

Infectious pancreatic necrosis (IPN) is a very serious viral disease in terms of its impact on production of Atlantic salmon, Salmo salar L., fry and post‐smolts. Post‐smolts of Atlantic salmon were injected with infectious pancreatic necrosis virus (IPNV) and cohabited with naive fish to produce natural infection. Cohabitant fish were sampled every 2 days, up to day 36 post‐infection (p.i.). From 90 cohabitant fish, 11 (12.2%) were positive by immunohistochemistry (IHC). The first detection of IPNV by IHC occurred on day 16 p.i. which coincided with the onset of mortality in this group. Besides the pancreas, the liver was found to be a key target organ for IPNV. For the first time, the virus was observed in the islets of Langerhans and in the kidney corpuscles of Stannius which suggests that the virus could affect the fish’s metabolism. The liver of two fish, which showed the most widespread presence of IPNV by IHC, had a pathology including focal necrosis and widespread presence of apoptotic hepatocytes, many of which did not stain for virus by IHC. Up‐regulation of cytokine gene expression was found only in the IHC‐positive (IHC+ve) fish and reflected the level of infection as determined by IHC positivity of the liver. In most fish, interferon (IFN), Mx, γIFN and γIP were up‐regulated in liver and kidney, while only IFN and Mx were up‐regulated in gill. IL1β and TNFα were not induced in any tissue. The gill showed variable levels of constitutive expression of IL1β and γIFN. The two fish with liver pathology had the highest level of IFN expression, especially relative to the level of Mx expression, in the liver compared with the other IHC+ve fish which did not have a liver pathology. The results suggest that following widespread infection of hepatocytes, the cells may over‐produce IFN, resulting in apoptosis of neighbouring cells with subsequent death from liver failure.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Vetch toxicosis in cattle grazing Vicia villosa ssp dasycarpa and V benghalensis

SUMMARY The epidemiological, clinical and pathological features of a disease syndrome in adult cattle grazing woolly-pod vetch (Vicia villosa ssp dasycarpa) or popany vetch (V benghalensis) are reported. Outbreaks of toxlcosis occurred between midwinter and midsummer in 3 dairy and 6 beef herds on the north coast of New South Wales, between 1982 and 1992. Friesian, Angus, Murray Grey, Guernsey and Hereford breeds were affected. Mean morbidity and case fatality rates in affected herds were 7% (65 of 889) and 69%, respectively. Signs of pruritic dermatitis, illthrift and death were associated with an eosinophilic granulomatous inflammation of many organs, particularly involving the renal cortex, dermis, myocardium, adrenal glands, lymph nodes and hepatic portal triads.     

Preliminary study of ovarian activity in fillies treated with a GnRH vaccine

Objective To investigate the effects of two doses (200 and 400 mg) of a water-soluble gonadotrophin-releasing hormone vaccine on the ovarian activity of 2-year-old fillies.
Design A controlled vaccination dose rate experiment.
Animals Six 2-year-old Australian Stock Horse fillies were randomly allocated to three treatment groups: unvaccinated controls, those receiving 200 mg of the vaccine and those receiving 400 mg of the vaccine.
Results Ovarian activity of the treated fillies was suppressed at the peak of the breeding season while that of the untreated controls continued normally. The control fillies displayed oestrous activity and behaviour. Suppression of ovarian activity occurred for 25 and 30 weeks in the 200 and 400 mg groups, respectively. These differences were not significant. Ovarian activity ceased 2 to 3 weeks after primary vaccination. Antibody titres were low (330) until after the booster immunisation when they rapidly peaked at 22,000 and 28,000 in the 200 mg and 400 mg groups, respectively. Plasma progesterone concentrations of the treated fillies remained below 3.18 nmol/L while GnRH was suppressed. The vaccine had no significant effect on plasma androstenedione concentrations. Recovery from the effect of the vaccine was associated with development of ovarian follicles, normal oestrous behaviour and ovulation. Three of the four treated fillies and one of the controls conceived during the next breeding season and foaled normally. All the treated fillies conceived and produced normal foals in the following two breeding seasons.
Conclusion Both dose rates suppressed ovarian function and prevented oestrous behaviour. These effects were reversible and the subsequent fertility of the vaccinated fillies was normal.     

Mother–Offspring Interactions in Zebu Cattle

The knowledge of the interaction between mother and offspring might contribute to enhance the welfare of the offspring and to improve the reproductive efficiency of the cow. However, there is still little information available about such interaction in some cattle breeds. A series of observational studies were set up, addressing the mother–offspring relationships of Nelore, Guzerat and Gyr cattle breeds. Firstly, the behaviour of cows and calves around the time of parturition was described, and then, the underlying factors that affect the calves' survival and development were studied. Special attention was given to the failure or delay in the first suckling. The results together are indicative of genetic variability for some studied variables, indicating the possibility of selection for calf vigour (using latency to stand up and latency to suckle as its indicators) and maternal ability (using percentage of time in contact with the calves), in spite of the estimates of heritability were low and presented high standard deviation for all variables. The individual variability in their suckling behaviour and the efficiency in first suckling cannot be explained by a single isolated underlying factor. By now, there are some results available, although there are many questions without answers. The field is still open for the development of future research.     

Changes in Testicular Interstitial Connective Tissue of Hamsters (Mesocricetus auratus) During Ageing and After Exposure to Short Photoperiod

The testicular interstitium of Syrian hamster (Mesocricetus auratus) was studied during ageing and in testicular regression after exposure to a short photoperiod, in relation to the interstitial cells and their connective tissue. This tissue was assessed histochemically using Masson's trichrome technique and the expression of Heat Shock Protein 47 (HSP‐47) and collagen IV (α5) was assessed in Leydig cells. Finally, an ultrastructural analysis of some cells of the testicular interstitium was made. Leydig cells were positive for HSP‐47 and collagen IV (α5). Ageing did not change the parameters studied while the short photoperiod altered the synthetic activity of Leydig cells. The positivity index of these cells for HSP‐47 was significantly higher in the regressed testis, but was lower for collagen IV (α5). During ageing no change were observed. Ultrastructural Leydig cells showed a discontinuous basal lamina that did not change during ageing. The basal lamina was not identified in Leydig cells regressed by exposure to a short photoperiod. In conclusion; the intertubular connective tissue suffers little change with age. By contrast, in the testis regressed after exposure to a short photoperiod the studied parameters related to the intertubular connective tissue were altered. These changes are probably related with the low synthetic activity of regressed Leydig cell.     

Using 20-gauge percutaneous peripheral catheters to reliably collect serial 4-mL blood samples from conscious dogs

Objective To determine the reliability of collecting serial 4-mL blood samples over 13 h using a 20-gauge, cephalic catheter in conscious dogs. Design Prospective, observational study. Animals Twelve (6 males, 6 females) healthy, neutered, lean dogs. Procedure Percutaneous placement of a 20-gauge, 1.1 × 30 mm intravenous over-the-needle catheter into the cephalic vein was performed in each dog on three occasions. Catheter patency was maintained by intermittent flushing with 0.9% saline and 1 IU/mL heparinised saline solutions. Blood samples (4 mL) were obtained at 10 time-points over 13 h, with close monitoring of the dogs and the indwelling catheters. Blood volume, resistance on aspirating blood sample (minimal or marked resistance) and the methods used to improve blood flow were recorded. Results A high proportion (93%) of blood sample collections achieved the required 4-mL volume, and the remaining samples were greater than 1.5 mL. Of the 358 collections via the cephalic catheter, 311 (87%) were obtained with ‘minimal resistance’. The remaining collections had ‘marked resistance’ (n=47) and were associated with a loose catheter in 11% (5/47) and of unknown cause in 89% (42/47). After ‘marked resistance’ had been encountered and the catheter was flushed with saline and heparin solutions, subsequent sampling with ‘minimal resistance’ was frequently possible from the same catheter. Conclusion Use of a percutaneous, 20-gauge intravenous cephalic catheter allowed reliable serial collection of 4-mL blood samples over 13 h in conscious dogs.