Effects of FGF10 on Bovine Oocyte Meiosis Progression,Apoptosis, Embryo Development and Relative Abundance of Developmentally Important Genes In Vitro

Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion‐related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22 h in TCM‐199 supplemented with 0, 2.5, 10 or 50 ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5 ng/ml FGF10 increased (p < 0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50 ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real‐time RT‐PCR showed a tendency of 50 ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10 ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.     

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA‐1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA‐1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo‐derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA‐1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA‐1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8–16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA‐1A revealed that it shares 91–98% identity with other mammalian sequences. It can be concluded that higher level of HSPA‐1A mRNA in IVP embryos in comparison with in vivo‐derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA‐1A gene could be used as a stress biomarker during pre‐implantation development.     

Vaccines for bluetongue

Isolation of 8 serotypes of bluetongue virus (BTV) in Australia has led to widespread debate on how to prepare for an outbreak of bluetongue disease and the type of vaccine best suited to control bluetongue in Australia. This article describes the vaccine options under consideration by research workers and animal health administrators. The most widely discussed options are live attenuated virus, killed virus and virus-like particles (VLP) generated by recombinant baculoviruses. Attenuated virus vaccines are cheap and easy to produce and are administered in a single dose. They replicate in sheep without causing significant clinical effects and provide protection against challenge with virulent virus of the same serotype. The possibility that insects could acquire vaccine virus by feeding on vaccinated animals and transmit it to sheep or cattle cannot be eliminated. This poses a risk because attenuated viruses are teratogenic if ewes are infected in the first half of pregnancy. In addition, vaccine virus replication in insects and ruminants may lead to a reversion to virulence. Killed virus vaccines have been shown to be efficacious in small laboratory trials and cannot be transmitted to other animals in the field, but are significantly more expensive to produce than attenuated viruses and require at least 2 doses with adjuvant to elicit an immune response. More work is needed to properly assess their effectiveness and determine their cost of production. Recombinant VLP contain the 4 major structural proteins of BTV but no nucleic acid. VLP are relatively easy to isolate, but it is unlikely that the purification methods currently used in laboratories will be adapted for use commercially. Despite the enthusiasm of recent years, little commercial progress appears to have been made. Although scientific research in Australia and overseas has provided a number of options for development of bluetongue vaccines, the decisions on which to use in an outbreak are complex and will require, not only consideration of factors discussed here, but also agreement from industry and government.     

Evaluation of the implementation of new traceability and food safety requirements in the pig industry in eastern Australia

Objectives   To evaluate the implementation and barriers to adoption, among pig producers, of a newly introduced traceability and food safety system in Australia.
Procedure   Implementation of the PigPass national vendor declaration (NVD) linked to an on-farm quality assurance (QA) program was evaluated in May and December 2007 at saleyards and abattoirs in New South Wales, Victoria and Queensland. Four focus group discussions with saleyard producers were held between April and July 2007.
Results   Implementation of the PigPass system in terms of accurate completion of the form and QA accreditation was higher at the export abattoir than at the regional saleyard at the first audit (P < 0.01). Implementation increased at the second audit at the abattoirs, but little change with time was observed at saleyards. Approximately half of the producers at saleyards used photocopied PigPass forms, made at least one error (>64%), and many vendors did not appear to be QA-accredited. During focus groups, producers expressed the view that PigPass implementation improved animal and product traceability. They identified the associated costs and a perceived lack of support by information providers as obstacles for adoption.
Conclusion   Improvement in the implementation of PigPass among producers marketing pigs at export abattoirs was observed during the 8-month period of the study. There is a need for a more uniform message to producers from government agencies on the importance of the PigPass NVD and QA and extension and education targeted toward producers supplying pigs to saleyards and domestic abattoirs to ensure compliance with the traceability requirements.     

Prevalence of proliferative enteritis on pig farms in Australia

SUMMARY Three surveys, undertaken to assess the prevalence of proliferative enteritis (PE) on pig farms in Australia and to investigate risk factors associated with clinical disease, indicated that PE was a common disease in pig farms. Forty of the 71 (56%) randomly-selected producers had either observed PE or had a veterinarian diagnose the disease in their herd during 1988 to 1990. A relatively low prevalence of the disease was recorded at veterinary diagnostic laboratories, and this suggested that diagnoses of PE were often not confirmed by histopathological examination of the intestines of affected pigs. Non-haemorrhagic PE occurred most often in six- to 24-week-old pigs, but was also reported in 52-week-old pigs. Proliferative haemorrhagic enteropathy usually affected pigs over 16 weeks of age, but was also reported in pigs as young as six weeks and as old as four years of age. A survey of pig-specialist veterinarians indicated that most veterinarians diagnosed PE based on clinical and gross pathological examination of affected pigs, without laboratory confirmation. There were difficulties associated with measuring the prevalence of PE among herds, including the effectiveness of antibacterials for its prevention and control, its subclinical nature and probable mis-diagnoses. This study highlighted the need for an ante-mortem diagnostic test to measure the prevalence of PE more accurately.     

Field Experiences on Post-cervical Artificial Insemination in the Sow

The present study was performed in order to evaluate the effects of post-cervical artificial insemination (post-CAI) in eastern European continental climate with multiparous sows. The sows were randomly allocated into two groups, and were AI by using CAI with 3 x 10(9) spermatozoa per dose (group 1, n = 859) or by post-CAI, using pooled semen with 1 x 10(9) spermatozoa per dose (group 2, n = 924). Wean-to-oestrus intervals, duration of oestrus, day 24 pregnancy rates, farrowing rates, and total pigs born were evaluated. Wean-to-oestrus intervals (CAI 114.3 +/- 4.1 h; post-CAI 115.2 +/- 5.2 h), duration of oestrus (CAI 64.1 +/- 4.1 h; post-CAI 65.0 +/- 5.2 h), day 24 pregnancy rates (CAI 90.2 +/- 1.7%; post-CAI 89.3 +/- 1.8%) and farrowing rates (CAI 88.1 +/- 2.3%; post-CAI 87.8 +/- 2.9%) did not differ significantly between CAI and post-CAI inseminated sows. The total number of pigs born differed significantly (p < 0.01) between the groups (CAI 12.3 +/- 1.1; post-CAI 10.2 +/- 0.9).     

Transmission studies of Hendra virus (equine morbilli-virus) in fruit bats, horses and cats

Objective To determine the infectivity and transmissibility of Hendra virus (HeV). Design A disease transmission study using fruit bats, horses and cats. Procedure Eight grey-headed fruit bats (Pteropus poliocephalus) were inoculated and housed in contact with three uninfected bats and two uninfected horses. In a second exper iment, four horses were inoculated by subcutaneous injection and intranasal inoculation and housed in contact with three uninfected horses and six uninfected cats. In a third experi ment, 12 cats were inoculated and housed in contact with three uninfected horses. Two surviving horses were inoculated at the conclusion of the third experiment: the first orally and the second by nasal swabbing. All animals were necropsied and examined by gross and microscopic pathological methods, immunoperoxidase to detect viral antigen in formalin-fixed tissues, virus isolation was attempted on tissues and SNT and ELISA methods were used to detect HeV-specific antibody. Results Clinical disease was not observed in the fruit bats, although six of eight inoculated bats developed antibody against HeV, and two of six developed vascular lesions which contained viral antigen. The in-contact bats and horses did not seroconvert. Three of four horses that were inoculated devel oped acute disease, but in-contact horses and cats were not infected. In the third experiment, one of three in-contact horses contracted disease. At the time of necropsy, high titres of HeV were detected in the kidneys of six acutely infected horses, in the urine of four horses and the mouth of two, but not in the nasal cavities or tracheas. Conclusions Grey-headed fruit bats seroconvert and develop subclinical disease when inoculated with HeV. Horses can be infected by oronasal routes and can excrete HeV in urine and saliva. It is possible to transmit HeV from cats to horses. Transmission from P poliocephalus t o horses could not be proven and neither could transmission from horses to horses or horses to cats. Under the experimental conditions of the study the virus is not highly contagious.     

Feeding of prohibited substances (swill) to pigs in Australia

Objective To assess current swill feeding legislation, swill feeding investigation practices by authorities and feeding practices of pig producers who trade via saleyards in eastern Australia in order to determine levels of understanding and conformance related to current swill feeding legislation. Method A three-tiered approach was undertaken to gather information on the feeding of prohibited substances (swill) to pigs in Australia. Firstly, a review of swill feeding legislation was undertaken to highlight the commonalities and inconsistencies between the various state and territory legislations in defining swill. Secondly, agricultural authorities were contacted in each state to gather information on swill feeding investigations undertaken in 2006. Finally, face-to-face interviews were conducted with 106 pig producers who traded pigs at one of six saleyards in eastern Australia to ascertain their knowledge of swill feeding and to determine the feeding practices of this sector of the industry. Results Areas of concern identified included (1) inconsistencies in the feedstuffs classed as ‘swill’ among states, (2) the number of producers who had been prosecuted for swill feeding in 2006 (n = 4 of 148 inspections), (3) the low knowledge base of producers who sell pigs at saleyards regarding swill feeding, and (4) the types of feedstuffs provided to pigs marketed at saleyards. Conclusion Our findings highlight the need for a consistent definition for ‘swill’ across Australian states and for improved awareness of swill feeding among producers, particularly those who market pigs at saleyards.