Effect of porcine circovirus type 2a or 2b on infection kinetics and pathogenicity of two genetically divergent strains of porcine reproductive and respiratory syndrome virus in the conventional pig model

To determine differences in infection kinetics of two temporally and genetically different type 2 porcine reproductive and respiratory syndrome virus (PRRSV) isolates in vivo with and without concurrent porcine circovirus (PCV) type 2a or 2b infection, 62 pigs were randomly assigned to one of seven groups: negative controls (n=8); pigs coinfected with a 1992 PRRSV strain (VR-2385) and PCV2a (CoI-92-2a; n=9), pigs coinfected with VR-2385 and PCV2b (CoI-92-2b; n=9), pigs coinfected with a 2006 PRRSV strain (NC16845b) and PCV2a (CoI-06-2a; n=9), pigs coinfected with NC16845b and PCV2b (CoI-06-2b; n=9), pigs infected with VR-2385 (n=9), and pigs infected with NC16845b (n=9). Blood samples were collected before inoculation and at day post-inoculation (dpi) 3, 6, 9 and 12 and tested for the presence of PRRSV antibody and RNA, PCV2 antibody and DNA, complete blood counts, and interferon gamma (IFN-γ) levels. Regardless of concurrent PCV2 infection, VR-2385 initially replicated at higher levels and reached peak replication levels at dpi 6. Pigs infected with VR-2385 had significantly higher amounts of viral RNA in serum on both dpi 3 and dpi 6, compared to pigs infected with NC16845b. The peak of NC16845b virus replication occurred between dpi 9 and dpi 12 and was associated with a delayed anti-PRRSV antibody response in these pigs. PCV2 coinfection resulted in significantly more severe macroscopic and microscopic lung lesions and a stronger anti-PRRSV IgG response compared to pigs infected with PRRSV alone. This work further emphasizes in vivo replication differences among PRRSV strains and the importance of coinfecting pathogens.     

Preliminary study on the activity of protease enzymes in Persian sturgeon (Acipenser persicus Borodin, 1897) larvae in response to different diets: effects on growth and survival

The specific activity of alkaline protease, trypsin and pepsin‐like enzymes was measured in yolk sac stage Acipenser persicus larvae and over a 1‐month feeding experiment using live Artemia nauplii (ND), formulated feed (FD) or mixed food (MD). Artemia nauplii group larvae showed significantly higher growth and survival during the first 15 days while FD larvae showed the lowest growth and survival. At day 30, MD larvae exhibited significantly higher growth than the ND group. Alkaline protease activity showed the lowest activity on day 15; the highest activity was observed in the MD group larvae. Pepsin‐like activity showed a drastic increase from day 1 to 5 in all treatments, but remained stable throughout the next 25 days, with the lowest and the highest activity in the FD treatment on day 10 and in the MD treatment on day 30 post‐feeding respectively. Trypsin‐like activity in group ND remained almost the same from day 5 to 30, whereas in groups MD and FD, it decreased significantly from day 10 to 30. The contribution of the naupliar proteases was moderate but effective. Additionally, better performance in Artemia fed sturgeon larvae may also be due to the structure and digestibility of proteins and the food intake stimulation by the nauplii.     

Efficient surveillance of pig populations using oral fluids

Currently virus surveillance in swine herds is constrained by the cost-effectiveness and efficiency of sampling methods. The objective of this study was to assess the value of using oral fluids collected by barn personnel as a method of surveillance based on PCR testing. Approximately 12,150 pigs in 10 wean-to-finish barns on 10 farms were monitored for the presence of porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), influenza A virus (IAV), and Torque teno virus genogroups 1 (TTV1) and 2 (TTV2) by sampling oral fluid specimens. Oral fluid samples were collected from 6 pens at each site starting at the time of pig placement (～3 weeks of age) and continuing thereafter at 2-week intervals for a period of 18 weeks. Data were analyzed both on a pen basis and barn basis. Overall, 508 (85%) samples were positive for PCV2, 73 (12%) for PRRSV, 46 (8%) for IAV, 483 (81%) for TTV2, and 155 (26%) for TTV1 during the study period. The estimated arithmetic means of the quantitative PCR-positive oral fluids for PCV2, PRRSV, and IAV were 1×10(4.62), 1×10(4.97), and 1×10(5.49)per ml. With a single exception, all barns were positive for PCV2 and TTV2 at every sampling point in the study. Virus detection varied among barns, particularly for IAV and PRRSV. The pen level, cumulative distribution of agent combinations between all 10 barns were statistically different. The most commonly observed patterns were PCV2+TTV2 (239 pen samples, 40%), PCV2+TTV1+TTV2 (88 pen samples, 15%), and PCV2 alone (66 pen samples, 11%). This "proof-of-concept" project showed that a variety of viruses could be detected either intermittently or continuously in pig populations and demonstrated that barn herd virus status is highly variable, even among barns in the same production system. Oral fluid sampling is a promising approach for increasing the efficiency and cost effectiveness of virus surveillance in swine herds.     

Evaluation of the influence of prostaglandin E2 on recombinant equine interleukin-1beta-stimulated matrix metalloproteinases 1, 3, and 13 and tissue inhibitor of matrix metalloproteinase 1 expression in equine chondrocyte cultures

OBJECTIVE: To determine the effects of prostaglandin E2 (PGE2) on recombinant equine interleukin (IL)-1beta-stimulated expression of matrix metalloproteinases (MMP 1, MMP 3, MMP 13) and tissue inhibitor of matrix metalloproteinase 1 (TIMP 1) in vitro. SAMPLE POPULATION: Cultured equine chondrocytes. PROCEDURE: Stationary monolayers of first-passage chondrocytes were exposed to graduated concentrations of PGE2 with or without a subsaturating dose (50 pg/ml) of recombinant equine IL-1beta (reIL-1beta) to induce expression of MMP 1, MMP 3, MMP 13, and TIMP 1, followed by RNA isolation and northern blotting. In subsequent experiments, gene expression was similarly quantified from mRNA isolated from cultures pretreated with phenylbutazone to quench endogenous PGE2 synthesis, followed by exposure to reIL-1beta and exogenous PGE2 (5 mg/ml) with appropriate controls. RESULTS: Exogenous PGE2 (10 mg/ml) significantly reduced reIL-1beta-induced expression of MMP 1, MMP 3, MMP 13, and TIMP 1. Abrogation of cytokine induction with this dose of PGE2 was comparable to that for dexamethasone (10(-5) M) control. Similarly, pretreatment with phenylbutazone, followed by exposure to relL-1beta and PGE2 (5 mg/ml), was associated with a reduced expression of the genes of interest, an effect that was significant for MMP 1, MMP 13, and TIMP 1. CONCLUSIONS AND CLINICAL RELEVANCE: The MMP and TIMP 1 are important mediators in the pathophysiologic events in osteoarthritis. The potential for physiologically relevant regulation of expression of these genes by PGE2 is a consideration in the use of drugs that inhibit prostanoid synthesis in the treatment of equine arthropathies.     

Short-range spatial variability of soil physico-chemical variables related to earthworm clustering in a neotropical gallery forest

In this study, we investigated the spatial distribution of an earthworm community together with the heterogeneity of selected soil properties in a gallery forest (GF) of the Colombian “Llanos”. We performed fine-scale spatial variability by intensively sampling 100 points distributed in the nodes of a regular grid with 5 m inter-sample distance. Non-parametric statistics were used and included SADIE analysis and partial Mantel test, in addition to geostatistics (semi-variograms) and correlogram computation. Our results indicated that the spatial distribution of earthworms was characterized by areas of presence (patches) and absence (gaps), although the general pattern was random at the scale of this study (<5 m), while soil physico-chemical characteristics showed a clumped spatial distribution. Contrary to previous results reported for the nearby savanna, a significant spatial association was found for two competing endogeic species Andiodrilus sp. and Glossodrilus sp. in the GF. Semi-variograms of soil environmental factors were adjusted to model families most commonly used (spherical and linear), and correlograms for earthworms showed significant positive and negative spatial autocorrelation for lag distances <15 m and >30 m, respectively. Partial Mantel test revealed specific significant relationships between soil variables and some species. The earthworm community of the GF displayed a random structure in a spatially clumped soil environment, and our results suggested that spatial distribution observed for some species could be the result of preferential selection of soil environmental factors. In other words, soil heterogeneity contributed to the formation of population patches for some earthworm species. The variability of suitable sites (resource availability patchiness) exerted an influence in the spatial distribution of earthworms at the scale used in this study, and we identified the spatial scale at which both environmental heterogeneity could influence and express earthworm impact on soil properties.     

Using occupancy models of forest breeding birds to prioritize conservation planning

As urban development continues to encroach on the natural and rural landscape, land-use planners struggle to identify high priority conservation areas for protection. Although knowing where urban-sensitive species may be occurring on the landscape would facilitate conservation planning, research efforts are often not sufficiently designed to make quality predictions at unknown locations. Recent advances in occupancy modeling allow for more precise estimates of occupancy by accounting for differences in detectability. We applied these techniques to produce robust estimates of habitat occupancy for a subset of forest breeding birds, a group that has been shown to be sensitive to urbanization, in a rapidly urbanizing yet biological diverse region of New York State. We found that detection probability ranged widely across species, from 0.05 to 0.8. Our models suggest that detection probability declined with increasing forest fragmentation. We also found that the probability of occupancy of forest breeding birds is negatively influenced by increasing perimeter-area ratio of forest fragments and urbanization in the surrounding habitat matrix. We capitalized on our random sampling design to produce spatially explicit models that predict high priority conservation areas across the entire region, where interior-species were most likely to occur. Finally, we use our predictive maps to demonstrate how a strict sampling design coupled with occupancy modeling can be a valuable tool for prioritizing biodiversity conservation in land-use planning.     

Modifications in the potato rhizosphere during infestations of <Emphasis Type="Italic">Globodera rostochiensis</Emphasis> and subsequent effects on the growth of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Two controlled environment experiments were conducted to explore the hypothesis that invasion and damage caused to potato roots by the potato cyst nematode Globodera rostochiensis might result in quantitative or qualitative changes in the release of root exudates to subsequently affect the growth of Rhizoctonia solani (AG3) in the potato rhizosphere. The growth of five R. solani isolates was compared on media amended either with root exudates from G. rostochiensis-infested or uninfested potato (cv. Désirée) plants at different time intervals after the introduction of the nematodes. In Experiment 1, the growth of R. solani was higher on medium amended with potato root exudates from G. rostochiensis-infested compared to uninfested plants, collected 4, 6, 8 and 12 days after the G. rostochiensis treatments were administered. Similarly, in Experiment 2, R. solani isolates grew faster on medium amended with potato root exudates from G. rostochiensis-infested than uninfested plants. This trend was particularly pronounced at the 12-day collection. At this time, 49% of the G. rostochiensis juveniles in roots were found to belong to the juvenile moults J2 and J3, indicating that root exudates were modified during the earlier stages of juvenile invasion. Carbohydrate analysis of root exudates indicated significantly higher levels of sucrose in root exudates from G. rostochiensis-infested than uninfested plants, whereas no significant differences were found in total nitrogen content. The results are discussed to help elucidate the mechanism behind the disease complex found between G. rostochiensis and R. solani in previous field research.     

Performance and welfare of broilers as affected by stocking density and zinc bacitracin supplementation

The influence of stocking density (16, 20 and 24 birds/m²) and zinc bacitracin (0 or 100 mg/kg) on the performance, carcass characteristics and some indirect welfare indicators of broilers were examined in a 35‐day trial. The weight gain, feed intake, liveability and carcass characteristics of broilers grown at densities of 16, 20 and 24 birds per m² were similar (P > 0.05). Over the whole trial period, however, feed per gain of birds at the highest density (24 birds/m²) was higher (P < 0.05) than those at the other two densities. Litter quality was poorer at high population densities, but there were no welfare implications as indicated by the lack of effect of density on gait scores and the incidence of hock and fat pad burns. Inclusion of zinc bacitracin improved weight gains and feed intake of broilers over the 35‐day trial period at all three stocking densities tested. Zinc bacitracin supplementation had no influence on the feed per gain of birds reared at densities of 16 and 20 birds/m², but improvements were observed at a density of 24 birds/m². The relative weights of lymphoid organs (spleen and bursa) were lowered (P < 0.05) with increasing density, suggesting a greater degree of stress on the birds. Zinc bacitracin supplementation had no influence on bursa weights of birds reared at densities of 16 and 20 birds/m², but increased the bursa weights of birds reared at a density of 24 birds/m². Overall, these data indicate that, with the removal of in‐feed antibiotics in the future, care should be taken when considering population densities of greater than 20 birds/m².