The effect on pathogenesis of Newcastle disease virus LaSota strain from a mutation of the fusion cleavage site to a virulent sequence

The principal molecular determinant of virulence of Newcastle disease virus (NDV) is the amino acid sequence at the fusion cleavage activation site. To extend the understanding of the role of the fusion cleavage activation site in NDV virulence, the pathogenesis in chickens of a lentogenic LaSota isolate and two infectious clones, NDFL and NDFLtag, were compared. NDFL is an infectious clone of a lentogenic NDV strain (LaSota E13-1), and NDFLtag is the infectious clone with the fusion cleavage site sequence mutated to the virulent motif. NDFL and NDFLtag were described by Peeters et al. The viruses were inoculated intraconjunctivally into groups of 4-wk-old white leghorn chickens and compared in a pathogenesis study for determination of disease causation (clinical signs of disease, gross lesions, histology, virus isolation, and serology) and viral distribution (presence of viral nucleoprotein and mRNA was detected by immunohistochemistry and in situ hybridization, respectively). The modification of the fusion cleavage activation site to the virulent motif in the infectious clone only slightly increased disease severity and viral distribution in the pathogenesis assessment, even though dramatically increased pathogenicity of NDFLtag was confirmed by standard pathogenicity index tests. The result, that the mutated fusion cleavage site of NDV-NDFLtag had only a small influence on pathogenesis in chickens compared to either E13-1 or NDFL, suggests that the pathogenic effects of NDV are not dependent on the fusion cleavage site alone.     

Plasma cortisol and hypothalamic monoamine responses in yellow perch <Emphasis Type="Italic">Perca flavescens</Emphasis> after intraperitoneal injection of lipopolysaccharide

The concentrations of monoamines in the hypothalamus were determined in yellow perch Perca flavescens before and after injection with lipopolysaccharide (LPS; 3 mg kg⁻¹ fish weight) or saline to test for the presence of neurochemical changes potentially associated with changes in plasma cortisol characteristic of intraperitoneal (ip) challenge with LPS. In the first experiment, yellow perch were injected with saline or LPS and the hypothalamus removed and plasma sampled before and at 0.5, 1.5, 3.0, and 6 h after injection. Plasma cortisol was elevated in both saline- and LPS-injected fish through 1.5 h after injection and returned to levels resembling pre-injection by 3 h after injection. Significantly higher amounts of cortisol in plasma from LPS-injected relative to saline-injected fish were observed 6 h following injection. A significant decrease relative to levels observed 0.5–3 h after handling was observed in serotonin concentrations at 6 h following LPS and saline injection with a concomitant increase in the ratio of 5-hydroxyindoleacetic acid:serotonin. In the second experiment, hypothalamic monoamines were sampled before and at 3, 6, 9, 12, and 24 h after injection with LPS or saline. Significant increases from pre-injection levels were observed in the ratio 5-hydroxyindoleacetic acid:serotonin at 9, 12, and 24 h after injection, but no differences were detected between LPS- and saline-injected fish. These results support a model linking serotonergic system activation following handling stress, but no correlations with the sustained elevations of plasma cortisol associated with inflammatory challenge were observed.     

A Method for In vivo Propagation of the Necrotizing Hepatopancreatitis Bacterium (NHPB) in Litopenaeus vannamei

The necrotizing hepatopancreatitis bacterium (NHPB) is difficult to preserve and has not been propagated in vitro. NHPB can be maintained continuously by simply adding specific pathogen free (SPF) shrimp to tanks containing NHPB‐infected shrimp. However, obtaining large amounts of highly concentrated infectious material free of contaminating bacterial flora for repeating challenge experiments was found to be exceedingly difficult using the current published methods. Therefore, a system was implemented using visible implant elastomer (VIE) tags to identify animals within a propagation tank by introduction group. Utilizing this method allowed for continuous reproduction of NHP within propagation tanks and provided researchers with access to infectious material allowing for consistent replication of challenge experiments and concentrated material for preservation.     

New strategies for the elimination of polio from India

The feasibility of global polio eradication is being questioned as a result of continued transmission in a few localities that act as sources for outbreaks elsewhere. Perhaps the greatest challenge is in India, where transmission has persisted in Uttar Pradesh and Bihar despite high coverage with multiple doses of vaccine. We estimate key parameters governing the seasonal epidemics in these areas and show that high population density and poor sanitation cause persistence by not only facilitating transmission of poliovirus but also severely compromising the efficacy of the trivalent vaccine. We analyze strategies to counteract this and show that switching to monovalent vaccine may finally interrupt virus transmission.     

Does Stocking Rate Manipulation Promote Pasture Sustainability in the Humid Tropics?

Stocking rate manipulation was examined as a means of improving plant diversity (as a measure of pasture sustainability and forage value) in a native grass pasture used for dairy production in the humid tropics of Veracruz, Mexico. Given that environmental impact reduces biotic diversity, plant phylogenetic and functional diversity should decline with increased stocking rate. Stocking rates of 2, 3, and 4 cows · ha^?1 and a rotational grazing plan of 3 d of occupation and 27 d of rest per pasture were applied continuously over 5 yr. Across 200 quadrats in each of two replicate paddocks per treatment, observed species richness, phylogenetic diversity (average taxonomic distinctness based on species presence/absence), and functional diversity (life-cycle duration and growth habit) were assessed. Most species were forb/herbs and forb/herb-subshrubs. Perennial species declined with increased stocking rate (F = 16.36, 0.05 > P > 0.02) while annual-perennial species increased (F = 76.88, 0.01 > P > 0.005); the proportion of annual species was least prominent and did not differ significantly. Observed species richness and phylogenetic diversity did not differ significantly with stocking rate. The correlation between functional diversity for life-cycle duration and phylogenetic diversity was significant and positive, suggesting that plant communities were predominantly assembled randomly from the surrounding species pool rather than through interspecies interactions acting to naturally filter immigrant species, thus leading to more opportunistic and undesired species. Although grazing pressure was not sufficient to alter indices or production measures, they did reveal shifts that may precede further pasture decline, indicating pasture sustainability was not being achieved. These rapid assessment methods permit monitoring for early warnings of reductions in pasture sustainability and forage quality for cattle.     

Concentration of 17α-Methyltestosterone in HormoneTreated Feed: Effects of Analytical Technique, Fabrication, and Storage Temperature

Feed treated with 17α-methyltestosterone (MT) is used to manipulate the gender of early tilapia fry. In the USA, hormone-treated feed is used by selected producers under an Investigational New Animal Drug (INAD) program. While monitoring the fabrication and on-farm use of this treated feed, concerns were raised about the high performance liquid chromatography (HPLC) analytical procedure for detecting MT in feeds, the incorporation of MT in feeds during fabrication, and effects of storage temperature on MT concentration. A series of experiments demonstrated that the analytical procedure for detecting MT in feeds lacked high precision, and, on average, biased results low. MT was uniformly mixed with feed by spraying an MT-alcohol solution on feed while it was blended in an industrial ribbon mixer. Alcohol volumes ranging from 15 mL/kg to 150 mL/kg were equally effective at dispersing MT in feed. The concentrations of MT in feeds consistently declined over time if the storage temperatures were 25 C or higher. Freezing preserved the MT in feed, and the refrigeration of feed fabricated to contain 60 mg MT/kg maintained acceptable MT concentrations during 6 mo of storage.