Experimental infection of pregnant gilts with swine hepatitis E virus

To determine the effect of swine hepatitis E virus (HEV) infection on pregnant gilts, their fetuses, and offspring, 12 gilts were intravenously inoculated with swine HEV. Six gilts, who were not inoculated, served as controls. All inoculated gilts became actively infected and shed HEV in feces, but vertical transmission was not detected in the fetuses. There was no evidence of clinical disease in the gilts or their offspring. Mild multifocal lymphohistiocytic hepatitis was observed in 4 of 12 inoculated gilts. There was no significant effect of swine HEV on fetal size, fetal viability, or offspring birth weight or weight gain. The offspring acquired anti-HEV colostral antibodies but remained seronegative after the antibodies waned by 71 days of age. Swine HEV infection induced subclinical hepatitis in pregnant gilts, but had no effect on the gilts' reproductive performance, or the fetuses or offspring. Fulminant hepatitis associated with HEV infection was not reproduced in gilts.     

The screening and evaluation of herbs and identification of herbal combinations with anti-viral effects on Newcastle disease virus

1. A total of 25 “heat-clearing and detoxifying” herbs used in Chinese medicine were investigated for their cytopathic effects on the growth of Newcastle Disease virus (NDV) in a chicken fibroblast cell line.

2. The 5 herbs with the highest virus inhibitory effects were Herba agastaches, Flos chrysanthemi indici, Rhizoma anemarrhenae, Astragalus root and Baikal skullcap root and these were used in herbal formulations. Anti-NDV activities of 4 formulations were tested on the growth of NDV in the DF-1 fibroblast cell line.

3. Formulation II, containing Baikal skullcap root, Astragalus root, Anemarrhena rhizome (1:1:2) and formulation IV containing Anemarrhena rhizome, Astragalus root and Flos chrysanthemi indici (1:1:1), which had strong anti-NDV activity in vitro, were used to determine the in vivo inhibitory effects of NDV-infection in chickens. After treatment with the two formulations serum IgY titres against NDV were improved, and morbidity was reduced in the NDV-infected chickens.

4. The results suggest that the components in formulations II and IV acted synergistically to improve resistance to Newcastle disease and provide a basis for the developing an anti-NDV herbal medicine.

     

A novel mutation of the CLCN1 gene associated with myotonia hereditaria in an Australian cattle dog

BACKGROUND: Heritable myotonia is a genetic muscle disorder characterized by slow relaxation of skeletal muscles. The main clinical signs are skeletal muscle stiffness, especially after vigorous contraction, and muscle hypertrophy. Muscle stiffness may be enhanced by inactivity, and often is relieved by exercise. Myotonia can be inherited in an autosomal dominant or recessive manner (Thomsen- or Becker-type myotonia, respectively). In mice, goats, Miniature Schnauzer dogs, and most affected humans, the disorder is caused by mutations in CLCN1, which encodes the skeletal muscle voltage-gated chloride channel, Cl1C-1. HYPOTHESIS: We hypothesized that an Australian Cattle Dog with generalized muscle stiffness and hypertrophy examined at the Ontario Veterinary College would have a mutation in the CLCN1 gene. ANIMALS: A pure-bred Australian Cattle Dog from Ontario, Canada, was used. METHODS: Based on clinical signs and electromyographic test results, a diagnosis of myotonia hereditaria was made, and a muscle biopsy was collected for genetic analysis. RESULTS: Sequence data obtained from the affected dog confirmed that it was homozygous for a single base insertion in the CLCN1 coding sequence. This mutation would result in a truncated ClC-1 protein being expressed, which, based on molecular evidence from other studies, would result in functionally compromised chloride conduction in the skeletal muscles of the animal. CONCLUSIONS AND CLINICAL IMPORTANCE: To the authors' knowledge, this report describes the Ist case of myotonia in an Australian Cattle Dog and represents the 1st non-Schnauzer canine myotonia to be genetically characterized. In addition, we developed a polymerase chain reaction-based genetic screen to detect heterozygotes with this mutation in the at-large Australian Cattle Dog population.     

Trade-offs among establishment success,stem morphology and productivity of underplanted Toona ciliata: Effects of nurse-species and thinning density

Toona ciliata (Australian red cedar), a valuable hardwood species, requires a nurse-tree overstorey to prevent damage from frost and drought in some regions of north-eastern Argentina. Different nurse-species offer different degrees of protection, resource competition, and thinning revenues. In addition, choice of nurse-species will influence trade-offs between T. ciliata establishment success, stem quality and productivity as well as revenue from intermediate and final harvests. We tested the effects of overstorey species and planting density on underplanted T. ciliata survival, stem morphology and growth. Initial stand densities of nurse-species Pinus taeda (625 stems/ha), P. elliottii × P. caribaea (625 stems/ha), and Grevillea robusta (833 stems/ha) were thinned by 0%, 25%, 50%, and 75% in three experiments, each using a randomized block design. Initial mortality, light availability and growth of T. ciliata, as well as growth of its nurse-trees, were measured before and two years after thinning. In the third year, we characterized T. ciliata stem morphology (straightness and height-to-diameter-ratio) and destructively harvested a subsample of T. ciliata in order to investigate T. ciliata biomass production and allocation patterns in response to overstorey species, thinning density and light availability. T. ciliata mortality was lower when planted under G. robusta than the pines, and increased in plots with greater exposure to solar radiation. T. ciliata productivity was also greater under G. robusta than under the pines, and was generally greatest where any of the nurse-species were thinned by 75%. Stem form was best when growing under P. taeda and in stands thinned by 50%, regardless of nurse-species. With the exception of early mortality, light levels appeared to have limited effects on T. ciliata growth dynamics. Neither growth rate nor stem morphology was strongly correlated with light availability. Total overstorey basal-area was highest in P. taeda stands and individual overstorey tree growth generally increased with decreasing stand densities, except for slower height growth of G. robusta in more open stands. Thinning operations resulted in sawn-wood revenue for the pine species only. Because stand productivity and stem quality were optimized under different nurse-species and thinning treatments, we conclude that the choice of nurse-species and thinning intensity will depend on management objectives and will change with market preferences and conditions.     

Evaluation of porcine ileum models of enterocyte infection by Lawsonia intracellularis

The early interaction of Lawsonia intracellularis with host cells was examined with the use of porcine ileum models. Two conventional swine were anesthetized, and ligated ileum loops were prepared during abdominal surgery. The loops were inoculated with 108 L. intracellularis or saline. After 60 min, samples of each loop were processed for routine histologic and electron microscopic study. Histologic and ultrathin sections of all the loops appeared normal, with no apposition of bacteria and host cells or bacterial entry events in any loop. Portions of ileum from a single gnotobiotic piglet were introduced as xenografts into the subcutis of each flank of 5 weaned mice with severe combined immunodeficiency disease. After 4 wk, 108 L. intracellularis were inoculated into each of 4 viable xenografts with a sterile needle; the other 3 viable xenografts received saline. Histologic and ultrathin sections of all the xenografts 3 wk after inoculation showed relatively normal porcine intestinal architecture, with normal crypts, crypt cell differentiation, and low villous structures; the xenografts treated with the bacteria also showed intracytoplasmic L. intracellularis within crypt and villous epithelial cells. Thus, entry of L. intracellularis into target epithelial cells and multiplication may not be sufficient alone to directly cause cell proliferation. A proliferative response may require active division of crypt cells and differentiation in conjunction with L. intracellularis growth.     

Performance of a foot-and-mouth disease virus reverse transcription-polymerase chain reaction with amplification controls between three real-time instruments.

The foot-and-mouth disease virus (FMDV) is a member of the picornavirus family, possessing an 8-kb single-stranded RNA genome of positive polarity. It is highly contagious among several livestock species and can lead to severe economic consequences, as evidenced by the UK outbreak in 2001. The usage of real-time polymerase chain reaction has facilitated rapid detection of FMDV. Several real-time PCR instruments are available with various capabilities, such as portability and high sample volume analysis. Primers and a dual-labeled TaqMan probe were optimized to detect a single, highly conserved 88-bp segment of the FMDV 3D (RNA polymerase) gene. To increase the confidence of the RT-PCR result, a positive amplification control was synthesized to detect potential false-positive results due to contamination if a wild-type virus is used as positive control. In addition, a preventative measure against false-negative results was developed in which endogenous beta actin mRNA is coamplified by RT-PCR. Assay performance was compared on the LightCycler1.2 (Roche), the SmartCyclerII (Cepheid), and the SDS 7900HT (ABI). These assays successfully identified the FMDV genome and beta actin mRNA from several sources of infected nasal and oral swabs, as well as probang samples.     

Experimental transmission of bovine anaplasmosis (caused by Anaplasma marginale) by means of Dermacentor variabilis and D. andersoni (Ixodidae) collected in western Canada

Canadian cattle are free of bovine anaplasmosis, with the exception of 4 isolated incursions since 1968, which were eradicated. It is not known why the disease has not become established in regions of Canada adjacent to the United States where it is endemic. To assess the vector competence of wild-caught ticks in cattle-rearing regions, Dermacentor variabilis and D. andersoni were collected in western Canada and fed on calves experimentally infected with Anaplasma marginale (St. Maries strain). The 2 tick species were equally competent in transmitting A. marginale to splenectomized calves, all 15 tick-exposed calves becoming infected. The prepatent periods in 13 calves ranged from 18 to 26 d and did not vary in relation to the numbers of ticks fed or the duration of transmission feedings. The unusually long prepatent periods in 2 calves (45 and 55 d) were probably due to concomitant Eperythrozoon infection. This study clearly demonstrated that tick species present in western Canada are competent vectors of bovine anaplasmosis. Potential barriers, including climate, must be considered in developing strategies to prevent A. marginale from becoming established in anaplasmosis-free regions.     

Adaptation of Asia-Pacific forests to climate change

Climate change is a threat to the stability and productivity of forest ecosystems throughout the Asia-Pacific region. The loss of forests due to climate-induced stress will have extensive adverse impacts on biodiversity and an array of ecosystem services that are essential for the maintenance of local economies and public health. Despite their importance, there is a lack of decision-support tools required to evaluate the potential effects of climate change on Asia-Pacific ecosystems and economies and to aid in the development of regionally appropriate adaptation and mitigation strategies. The project Adaptation of Asia-Pacific Forests to Climate Change, summarized herein, aims to address this lack of knowledge and tools and to provide support for regional managers to develop effective policy to increase the adaptive capacity of Asia-Pacific forest ecosystems. This objective has been achieved through the following activities: (1) development of a high-resolution climate downscaling model, ClimateAP, applicable to any location in the region; (2) development of climate niche models to evaluate how climate change might affect the distribution of suitable climatic conditions for regionally important tree species; (3) development and application of forest models to assess alternative management strategies in the context of management objectives and the projected impacts of climate change; (4) evaluation of models to assess forest fire risk and the relationship between forest fire and climate change; (5) development of a technique to assess ecosystem carbon storage using LiDAR; and (6) evaluation of how vegetation dynamics respond to climate change using remote sensing technology. All project outputs were developed with a focus on communication and extension to facilitate the dissemination of results to regional forest resource managers to support the development of effective mitigation and adaptation policy.