Effect of dietary vitamin E supplementation and feeding period on pork quality

Feeding increased levels of dietary vitamin E can inhibit lipid oxidation. The aim of this study was to investigate the effect of levels of dietary alpha-tocopherol acetate (VE) and feeding duration on meat quality and lipid oxidation. Eighty-one pigs were allocated to 1 of 3 diets containing 40, 200, or 400 IU of VE/kg of feed, and each diet group was divided into 3 feeding periods (3, 6, or 9 wk). Carcass characteristics and meat quality were evaluated. Oxidative stability of fresh and cooked pork patties and pork chops was determined after chilled or frozen storage. Increasing dietary concentrations of VE did not affect any growth performance parameter. Drip loss, however, decreased (P < 0.05) with increased dietary VE levels. Moreover, an increased duration of VE feeding improved (P < 0.05) pH and drip loss. Less lipid oxidation (P < 0.05) was detected in fresh ground pork from pigs fed greater concentrations of VE after 4 d of storage. A greater (P < 0.05) resistance to oxidation in cooked ground pork was observed in pigs fed 200 or 400 IU of VE/kg at 2 and 6 d of storage. Fresh and cooked pork patty oxidation decreased (P < 0.05) linearly as feeding duration increased from 3 to 9 wk. After 6 mo of freezer storage, lipid oxidation of pork chops from pigs fed 200 or 400 IU of VE/kg was lower (P < 0.05) than for pigs fed 40 IU of VE/kg. Likewise, lipid oxidation of pork chops of pigs fed VE for an extended period of time (6 wk) was lower (P < 0.05) after 9 mo of storage. Fatty acid profiles of neutral lipid fraction of the LM became more unsaturated (P < 0.05) with added VE to the feed. These results indicate an increased intake of dietary VE concentration, and prolonged feeding of VE can improve drip loss and reduce lipid oxidation in ground pork and pork chops. This study suggests that supplementation with 200 IU of VE/kg of feed for 6 wk before market is beneficial in improving lipid stability and pork quality.     

Metabolism of isometamidium in hepatocytes isolated from control and inducer-treated rats

Little is known about the metabolism and mechanism of action of the trypanocide, isometamidium (ISM), the major drug used for prophylaxis of trypanosomiasis. We have investigated its metabolism and distribution in isolated rat hepatocytes using liquid chromatography-mass spectrometry and confocal laser scanning microscopy (CLSM). Two putative metabolites were formed, which were proposed to be a mono-acetyl derivative and an oxidized metabolite (SII). This is the first demonstration of the hepatic metabolism of ISM, as previous in vivo studies were hampered by dose-limiting toxicity and insensitive analytical methods. The intrinsic fluorescence of the drug enabled its intracellular uptake to be followed by CLSM. It is taken up rapidly into the nucleolus, nuclear membrane and endoplasmic reticulum within 5 min, and retained in the nucleus for at least 24 h. Persistent binding of ISM to cellular macromolecules may contribute to its prophylactic effect in vivo. Pretreatment of rats with 3-methylcholanthrene, phenobarbitone (PB) or the widely used pyrethroid pesticide, deltamethrin, resulted in an increase in metabolism of ISM to the proposed SII after 1 h incubation with hepatocytes. 3-methylcholanthrene was the most potent inducer, causing a maximal 19.5-fold induction of SII formation after exposure of hepatocytes to ISM for 1 h compared with formation by control hepatocytes. In comparison, at the 1 h timepoint deltamethrin pre-treatment caused a 10.2-fold induction, and PB only 8.2 fold.     

Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice

The Mycoplasma hyopneumoniae ribonucleotide reductase R2 subunit (NrdF) gene fragment was cloned into eukaryotic and prokaryotic expression vectors and its immunogenicity evaluated in mice immunized orally with attenuated Salmonella typhimurium aroA CS332 harboring either of the recombinant expression plasmids. We found that NrdF is highly conserved among M. hyopneumoniae strains. The immunogenicity of NrdF was examined by analyzing antibody responses in sera and lung washes, and the cell-mediated immune (CMI) response was assessed by determining the INF-gamma level produced by splenocytes upon in vitro stimulation with NrdF antigen. S. typhimurium expressing NrdF encoded by the prokaryotic expression plasmid (pTrcNrdF) failed to elicit an NrdF-specific serum or secretory antibody response, and IFN-gamma was not produced. Similarly, S. typhimurium carrying the eukaryotic recombinant plasmid encoding NrdF (pcNrdF) did not induce a serum or secretory antibody response, but did elicit significant NrdF-specific IFN-gamma production, indicating induction of a CMI response. However, analysis of immune responses against the live vector S. typhimurium aroA CS332 showed a serum IgG response but no mucosal IgA response in spite of its efficient invasiveness in vitro. In the present study we show that the DNA vaccine encoding the M. hyopneumoniae antigen delivered orally via a live attenuated S. typhimurium aroA can induce a cell-mediated immune response. We also indicate that different live bacterial vaccine carriers may have an influence on the type of the immune response induced.     

Experimental comparison of aerial larvicides and habitat modification for controlling disease-carrying Aedes vigilax mosquitoes

BACKGROUND: Microbial and insect‐growth‐regulator larvicides dominate current vector control programmes because they reduce larval abundance and are relatively environmentally benign. However, their short persistence makes them expensive, and environmental manipulation of larval habitat might be an alternative control measure. Aedes vigilax is a major vector species in northern Australia. A field experiment was implemented in Darwin, Australia, to test the hypotheses that (1) aerial microbial larvicide application effectively decreases Ae. vigilax larval presence, and therefore adult emergence, and (2) environmental manipulation is an effective alternative control measure. Generalised linear and mixed‐effects modelling and information‐theoretic comparisons were used to test these hypotheses. RESULTS: It is shown that the current aerial larvicide application campaign is effective at suppressing the emergence of Ae. vigilax, whereas vegetation removal is not as effective in this context. In addition, the results indicate that current larval sampling procedures are inadequate for quantifying larval abundance or adult emergence. CONCLUSIONS: This field‐based comparison has shown that the existing larviciding campaign is more effective than a simple environmental management strategy for mosquito control. It has also identified an important knowledge gap in the use of larval sampling to evaluate the effectiveness of vector control strategies. Copyright © 2011 Society of Chemical Industry     

Generation of leaf shape through early patterns of growth and tissue polarity

A major challenge in biology is to understand how buds comprising a few cells can give rise to complex plant and animal appendages like leaves or limbs. We address this problem through a combination of time-lapse imaging, clonal analysis, and computational modeling. We arrive at a model that shows how leaf shape can arise through feedback between early patterns of oriented growth and tissue deformation. Experimental tests through partial leaf ablation support this model and allow reevaluation of previous experimental studies. Our model allows a range of observed leaf shapes to be generated and predicts observed clone patterns in different species. Thus, our experimentally validated model may underlie the development and evolution of diverse organ shapes.     

Influence of urban edges on stress in an arboreal mammal: a case study of squirrel gliders in southeast Queensland, Australia

There is growing recognition that ecological research must expand its focus beyond inference based on pattern-process relationships to the direct measurement of ecological and physiological processes. Physiological assessment is important because vertebrates cope with unpredictable and noxious stimuli by initiating a stress response. However, an over-activation of the acute stress response by numerous novel and potentially stressful anthropogenic pressures, including those associated with urban edges, has the potential to generate chronic stress and a greater susceptibility to disease, reduce fecundity and survivorship. An individual??s physiological response to edge habitats with varying degrees of contrast to the adjacent disturbed urban matrix (e.g. major vs. minor roads), may provide insight into their survival likelihood in fragmented urban landscapes. Although demographic changes in wildlife resulting from urbanization have been documented, only recently have physiological consequences been examined. We addressed this problem using a case study of the squirrel glider (Petaurus norfolcensis) in the fragmented urban landscape of southeast Queensland, Australia. Hair samples were used to enable a comparison of hair cortisol levels in individual squirrel gliders, providing an indication of potential stress. We applied a linear mixed-effect modeling approach clustered by patch to quantify the influence of site-level habitat factors and relative abundance comparative to edge contrast on hair cortisol levels. We found that edge type had a strong positive effect on hair cortisol levels; but this depended on the availability of abundant nest hollows at a site. We conclude that individual hair cortisol concentration, providing an index of stress, was lowest in interior habitats and highest in edge habitats adjacent to major roads. Furthermore, gliders occupying low edge contrast habitats adjacent to residential areas and minor roads, and containing abundant tree nest hollows, had low-moderate hair cortisol levels. This highlights the potential importance of these habitats for the conservation of arboreal mammals such as the squirrel glider in urban landscapes.     

Genetic variation in the 5′UTR of the KRT2.13 gene of sheep

KRT2.13 is a type II keratin wool intermediate filament (IF) protein. Extensive variation was revealed in the 5′ untranslated region (UTR) of the ovine KRT2.13 gene (KRT2.13) using polymerase chain reaction – single strand conformational polymorphism (PCR‐SSCP) analysis. Nine unique PCR‐SSCP patterns were obtained with individual sheep having either one (homozygous), or a combination of two (heterozygous) of these patterns. Seven of the amplicons that produced the apparently homozygous patterns were successfully sequenced (GenBank FJ217670 – FJ217676), revealing eight single nucleotide insertions, 10 single nucleotide substitutions, a nucleotide deletion and a 16 nucleotide insertion that occurred in only one of the sequences. The seven sequences showed between 85% and 95% homology to the previously identified KRT2.13 sequence (GenBank X72379). This study emphasizes the power of PCR‐SSCP analysis in genotyping, as this extensive variation was found in only 100 sheep, of a variety of breeds. Since variation in the 5′UTR of genes may affect their expression, this genetic variation needs to be further studied to establish its role if any, in influencing gene expression and consequently wool traits.     

Epitope-tagged insulin-like growth factor-I expression in muscle

Development of a recombinant insulin like growth factor I (IGF-I) that is distinguishable from its endogenous counterpart would provide a powerful tool for delineating the role of IGF in myogenesis. Therefore, the objective of this study was to create an epitope-tagged IGF-I that retains biological activity and determine whether expression of this construct is possible in muscle tissue following direct DNA injection. Expression vectors were created that encoded porcine IGF-I containing a T7 (11-amino acid) epitope-tag (TIGF). Immunoreactivity of the purified recombinant TIGF was confirmed using monoclonal antibodies. Biological activity was evaluated by examining differentiation of myoblasts cultured with TIGF or transfected with TIGF plasmid DNA. Addition of purified TIGF to myoblast cultures stimulated (P < 0.05) muscle creatine kinase levels similar to insulin (10(-5) M). Likewise, transfection of L6A1 with TIGF DNA hastened (P < 0.01) differentiation compared to control pcDNA-transfected myoblasts. The integrity of the recombinant protein was confirmed using a sandwich-configured enzyme linked immunosorbent assay. Finally, recombinant TIGF DNA was injected in porcine muscle and the ability to detect TIGF protein was evaluated. TIGF expression was detected in muscle fibers of injected porcine muscle. These data show that a T7 amino acid tag placed on the amino terminus of the IGF-I protein remains intact during processing and does not interfere with the biological activity of the molecule. Use of this DNA construct is an excellent tool for investigating the role of IGFs in control muscle development and provides a model to investigate other regulators of animal growth.     

VERTEBRAL HEART SCORES IN EIGHT DOG BREEDS

The vertebral heart score (VHS) measurement is commonly used to provide a more objective measurement of cardiomegaly in canines. However, several studies have shown significant breed variations from the value previously established by Buchanan and Bücheler (9.7 ± 0.5). This study describes VHS measurements in Pug, Pomeranian, Yorkshire Terrier, Dachshund, Bulldog, Shih Tzu, Lhasa Apso, and Boston Terrier dog breeds. Dogs with two or three view thoracic radiographs, no subjective radiographic evidence of cardiomegaly, and no physical examination findings of heart murmurs or gallop rhythms were included in the study. The Pug, Pomeranian, Bulldog, and Boston Terrier groups were found to have a VHS significantly greater than 9.7 ± 0.5 (P < 0.00001, P = 0.0014, P < 0.0001, P < 0.00001, respectively). Body condition score (BCS) was found to have a significant effect on the VHS of Lhasa Apso group. Anomalous vertebrae in the thoracic column were associated with a significant increase in VHS of the Bulldog (P = 0.028) and Boston Terrier (P = 0.0004) groups. Thoracic depth to width ratio did not have a significant effect on VHS.