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.     

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).     

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.     

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.     

Effects of foam fractionation and chemical disinfection on the removal of different microalgae cultures

The recent development of recirculation aquaculture systems (RAS) potentially allows more sustainable and controlled rearing conditions. However, control of suspended microparticles and microalgae in RAS is challenging, as uncontrolled blooms of toxic algae or heterotrophic dinoflagellates can have catastrophic impacts on the production of fish. In this study, we tested the potential of protein skimmers to remove microalgae. In 100 L batch tests, protein skimmers were tested separately and in combination with ozone (O₃), ultraviolet (UV) or hydrogen peroxide (H₂O₂). Three different and distinct microalgae cultures were tested with densities from 30,000 to 120,000 algae cells/ml in a triplicated experimental setup. Trial 1 included twelve 24‐hr replicated tests where protein skimmers with air or with two levels of ozone (low and high O₃ exposure) were compared. The protein skimmer with air alone had a limited effect on the removal of microalgae compared to the untreated control algae tanks. When ozone and protein skimmers were combined, a clear additive effect was found, and all added microalgae were removed. Low O₃ dosage and protein skimmers completely removed the algae cultures within 12 hr, while more than 95% of the algae were removed within 7 hr when a higher ozone dose was used. The second trial compared the removal capacity of protein skimmers in combination with UV, H₂O₂ and O₃. These experiments showed no to limited additive effect of UV combined with protein skimming, but significantly increased removal efficiency (270% and 1,300%, respectively) was found when H₂O₂ and ozone were combined with a protein skimmer. The study showed an algae species‐specific response to a protein skimmer with ozonation and provided information on the transition from reactivity and consumption to accumulation of ozone‐produced oxidants.     

A genome-wide association study identifies IL23R as an inflammatory bowel disease gene

The inflammatory bowel diseases Crohn's disease and ulcerative colitis are common, chronic disorders that cause abdominal pain, diarrhea, and gastrointestinal bleeding. To identify genetic factors that might contribute to these disorders, we performed a genome-wide association study. We found a highly significant association between Crohn's disease and the IL23R gene on chromosome 1p31, which encodes a subunit of the receptor for the proinflammatory cytokine interleukin-23. An uncommon coding variant (rs11209026, c.1142G>A, p.Arg381Gln) confers strong protection against Crohn's disease, and additional noncoding IL23R variants are independently associated. Replication studies confirmed IL23R associations in independent cohorts of patients with Crohn's disease or ulcerative colitis. These results and previous studies on the proinflammatory role of IL-23 prioritize this signaling pathway as a therapeutic target in inflammatory bowel disease.     

Genomic regions associated with ventro-cranial chronic pleuritis in pig

Ventro‐cranial chronic pleuritis can be a result of pleuropneumonia and enzootic pneumonia. These diseases cause severe losses in intensive pig production worldwide, but host resistance is difficult to breed for. It could be beneficial to use marker‐assisted selection, and a step towards this is to identify genomic regions associated with the trait. For this purpose, 7304 pigs from 11 boar families were analysed for associations between single nucleotide polymorphisms and ventro‐cranial chronic pleuritis. The pigs were genotyped by the use of the iSelect Custom 7 K porcine SNP Chip. Quantitative trait loci (QTL), significant at the chromosome‐wide level, were identified on Sus scrofa chromosomes (SSC) 2, 4, 11, 12 and 13 in four different boar families. The QTL on SSC 4 in family G was also significant at the genome‐wide threshold according to Bonferroni correction. We have identified a number of candidate genes, but the causative mutations still need to be identified. Markers closely associated with the resistance traits have a strong potential for use in breeding towards animals with improved characteristics concerning ventro‐cranial chronic pleuritis.     

Protein heterogeneity of bovine β-casein in Danish dairy breeds and association of rare β-casein F with milk coagulation properties

A proteomic approach was applied to characterize genetic variation and relative protein content of β-casein (β-CN) in milk from two minor native dairy breeds in Denmark; Red Danish 1970 and Jutland cows in comparison to the major dairy breeds; Danish Holstein and Danish Jersey. Genetic β-CN variants were related to the relative concentration of β-CN. Furthermore, a very high frequency of variant F in the native breeds was found. This was remarkable as this variant is only found in a very low frequency in the major Danish breeds. Novel associations were found between β-CN variant F and milk coagulation properties in the native breeds, which is an important finding for understanding the genetic background for non- and poor coagulating milk and potential exploitation of β-CN variants in selective breeding programs. Furthermore, in Red Danish 1970 a very high prevalence of non-coagulating milk was found (18%).