Isolation and Differentiation of Mesenchymal Stem Cells From Bovine Umbilical Cord Blood

Currently, mesenchymal stem cells (MSCs) are used in veterinary clinical applications. Bone marrow and adipose tissue are the most common sources of stem cells derived from adult animals. However, cord blood which is collected non‐invasively is an alternative source of stem cells other than bone marrow and adipose tissue. Moreover, high availability and lower immunogenicity of umbilical cord blood (UCB) haematopoietic stem cells compared to other sources of stem cell therapy such as bone marrow have made them a considerable source for cell therapy, but MSCs is not highly available in cord blood and their immunogenicity is poorly understood. In this study, the cells with spindle morphology from 7 of 9 bovine UCB samples were isolated and cultured. These mesenchymal stromal cells were successfully differentiated to osteocytes, chondrocytes and adipocytes. In addition, Oct‐4 and SH3 were determined by RT‐PCR assay. It is the first report of isolation, culture, characterization and differentiation of bovine umbilical stem cells.     

Phenotypic and genotypic characterisation of Pasteurella multocida isolated from pigs at slaughter in New Zealand

AIMS: To examine pigs at slaughter in New Zealand for the presence of Pasteurella multocida, and to determine for isolates, their biochemical profiles, somatic and capsular types, and the presence or absence of the HSB and toxA genes, associated with haemorrhagic septicaemia (HS) and progressive atrophic rhinitis (PAR), respectively.

METHODS: Swabs from 173 lungs, 158 palatine tonsils and 82 nasal passages of pigs at two abattoirs in New Zealand were cultured for P. multocida using conventional techniques, and isolated colonies were subjected to biochemical tests for identification of biovars. Somatic serotyping was conducted using an agar gel immunodiffusion (AGID) test. Polymerase chain reaction (PCR) assays were used to confirm phenotypic identification of colonies using species-specific primers, capsule type using serogroup-specific primers and multiplex PCR, and to test for the presence of HSB and toxA genes.

RESULTS: Pasteurella multocida was isolated from 11/173 (6.4%) lung, 32/158 (20.2%) palatine tonsil and 5/82 (6.1 %) nasal swab samples, a total of 48 isolates from 413 samples (11.6%). Isolation rates per farm ranged from 1–53% of tissue samples collected from pigs 5–6 months of age. On phenotypic characterisation, isolates were allocated to seven main biovars, viz 1, 2, 3, 5, 9, 12, and a dulcitol-negative variant of Biovar 8, the majority (30/48) being Biovar 3. Of the 42 isolates for which somatic serotyping was conducted, 10% were Serovar 1, 79% were Serovar 3, 2% were Serovar 6,1, 2% were Serovar 12, and 7% could not be typed. All 48 isolates were confirned as P. multocida using a species-specific PCR. In the capsular multiplex PCR, 92% of isolates were Capsular (Cap) type A, 2% were Cap D, and 6% could not be typed. None of the samples were positive for the HSB or toxA genes.

CONCLUSION: Serovars or capsular types of P. multocida associated with HS or PAR in pigs were not detected. Establishment of species-specific, capsular and toxin PCR assays allowed the rapid screening of isolates of P. multocida, while serotyping provided an additional tool for epidemiological and tracing purposes.     

Detection of Mycoplasma conjunctivae in sheep affected with conjunctivitis and infectious keratoconjunctivitis

AIM: To determine the aetiolog y of a recurring and severe form of infectious keratoconjunctivitis (IKC) in sheep.

METHODS: Five sheep flocks that had experienced a severe form of IKC were examined. Clinical history, conjunctival swabs and blood samples were collected from affected animals. Culture for bacteria, and also specifically for Mycoplasma and Chlamydophila spp, and detection of Mycoplasma conjunctivae DNA by polymerase chain reaction (PCR) were attempted. Serum samples were tested for antibodies to M. agalactiae, M. capricolum, M. conjunctivae and Chlamydophila spp.

RESULTS: Mycoplasma conjunctivae DNA was detected using PCR in 3/5 flocks, and in all flocks antibodies to M. conjunctivae were detected in sera. A pure growth of Branhamella ovis was cultured from conjunctival swabs from a small proportion of sheep in two flocks. No other pathogens were detected.

CONCLUSIONS: This investigation demonstrated that M. conjunctivae was a primary pathogen causing severe IKC in sheep, and is the first report of detection of this organism in sheep in New Zealand. Introduction of clinically normal carrier sheep appeared to have caused the outbreaks.     

Real-time ultrasonography for pregnancy diagnosis and foetal ageing in fallow deer

SUMMARY Three groups of 15 to 17 adult fallow does with some additional yearling does in 2 of the groups were treated to synchronise oestrous cycles, and mated. All does were scanned by ultrasound at 4 weeks of gestation and at weekly intervals from week 7 to week 14 of gestation. Growth rates of 13 foetal and uterine characters, which have been used for ageing foetuses of red deer, were similar for adult and yearling does and among the 3 groups. Transrectal ultrasound scanning was a reliable and accurate means of detecting pregnancy and of ageing foetuses of fallow deer during weeks 7 to 17 of pregnancy.     

Using GIS to measure changes in the temporal and spatial dynamics of forestland: experiences from north-west Spain

Forestry variables are usually calculated at a forest managementunit scale. However, a region's forestry sector is affectedby various other factors that interact over space and time,many of which are not directly associated to silvicultural activitiesbut nonetheless play an important part in its development froma socio-economic or environmental point of view. To understanda region's forestry dynamics, and especially to predict itsfuture tendencies, we must include all the necessary variablesin a single database, calculated for spatial units that arestable over time and adequate for planning purposes. In ourstudy, we developed a Forest Geographic Information System forGalicia called ‘SIFGa’. We used it to examine 310variables describing the environment, population tendencies,land tenure and forest management in the Spanish autonomousregion of Galicia, at both council and parish levels. Resultsreveal the connections between our variables, which reflectthe changes the regional forestry sector has experienced inthe past, and explain its current situation. They also confirmthe heterogeneity of forestry in the area and the need to adaptforest-planning strategies to each study unit, as well as tothe entire region.     

A cross-sectional pilot study to estimate the prevalence of and risk factors for leptospirosis in South-Western Victorian dairy herds, 2017

Leptospirosis is a zoonosis, found worldwide, affecting many species of animals. We conducted a cross-sectional study to estimate the prevalence of Leptospira borgpetersenii sv Hardjo and Leptospira interrogans sv Pomona in cattle in dairy herds in South-Western Victoria, Australia. Fifty-three herds were enrolled in the study. Urine samples were collected from 15 late-lactation cows in each herd. A questionnaire was provided to herd managers at the time of each herd visit, asking them to describe the methods they used for controlling leptospirosis, including vaccination. Urine samples were pooled at the herd level and tested for leptospira spp. using real time PCR. Urine samples from individual cows within the positive pooled samples were then tested for Leptospira Hardjo and Leptospira Pomona using qPCR. Four of the 53 herds showed positive leptospirosis results giving an apparent prevalence of 8 (95% CI 2–18) leptospira-positive herds per 100 herds at risk. Based on the 53 completed questionnaires, leptospirosis vaccination programs were not compliant with label directions in 36 of the 52 vaccinated herds: 69 (95% CI 55–81) of 100 herd managers that routinely vaccinated for leptospirosis did not comply with label directions. One herd was completely unvaccinated. Based on our findings, we estimate that approximately 10% of dairy farms in South-Western Victoria are likely to be infected with leptospirosis. While most herds are vaccinating for leptospirosis, most are not doing so according to label directions. We conclude that herd managers need to be better educated regarding leptospirosis vaccination programs.     

Bovine ephemeral fever

Ephemeral fever remains a viral disease of considerable importance to many countries including Australia. The virus has been only partly characterised and still awaits final classification. Although BEF virus was first thought to contain 6 structural proteins there is increasing evidence to suggest that it contains the 5 proteins characteristic of the Rhabdoviridae. Although BEF is thought to be arthropod borne, the vector has yet to be identified but it is clear from the distribution of BEF that more than one vector is capable of transmitting the disease. Despite rigorous investigation of the clinical signs and the pathology of ephemeral fever, little progress has been made on the pathogenesis of the disease. This has been partly due to the difficulty of propagating BEF virus in vitro and the inability to define the site of replication. However, there is mounting evidence to suggest that BEF is immunopathologic in nature and that the clinical expression of the disease is influenced by the release of one or more mediators of inflammation. The disease is characterised by a number of haematological and biochemical changes and early and prolonged treatment with phenylbutazone is capable of reversing a number of these changes. The intravenous administration of calcium can now be considered a justifiable addition to the treatment regimen together with prolonged phenylbutazone therapy. The vaccines currently available are prepared from either live attenuated or killed virus and may be less than reliable. There appears to be a need for a reliable, inexpensive, cold-chain independent alternative vaccine.