DNA vaccination of the American crow (Corvus brachyrhynchos) provides partial protection against lethal challenge with West Nile virus

The New York 1999 strain of West Nile virus (WNV) is nearly 100% fatal in the American crow (Corvus brachyrhynchos). We evaluated four WNV vaccine formulations in American crows, including intramuscular (i.m.) DNA vaccine, i.m. DNA vaccine with adjuvant, orally administered microencapsulated DNA vaccine, and i.m. killed vaccine. Neutralizing antibodies developed in approximately 80% of crows that received the DNA vaccine i.m. (with or without adjuvant), and in 44% that received the killed vaccine. However, no crows that received the oral microencapsulated DNA vaccine or the placebo developed WNV antibodies. All crows were challenged 10 wk after initial vaccination. No unvaccinated crows survived challenge, and survival rates were 44% (i.m. DNA vaccine), 60% (i.m. DNA vaccine with adjuvant), 0% (oral microencapsulated DNA vaccine), and 11% (killed vaccine). Peak viremia titers in the birds that survived were significantly lower as compared to titers in birds that died. Parenteral administration of a WNV DNA vaccine was associated with reduced mortality but did not provide sterile immunity.     

Molecular characterization of Sarcocystis spp. as a cause of protozoal encephalitis in a free-ranging black bear

A free-ranging juvenile male black bear (Ursus americanus), found dead in Alberta, Canada, had severe nonsuppurative encephalitis. Lesions in the brain were most severe in the gray matter of the cerebral cortex, and included perivascular cuffs of lymphocytes and plasma cells, areas of gliosis that disrupted the neuropil, and intralesional protozoan schizonts. The left hindlimb had suppurative myositis associated with Streptococcus halichoeri. Immunohistochemistry and molecular analyses (PCR and sequencing of 4 discriminatory loci: 18S rDNA, ITS-1 rDNA, cox1, rpoB) identified Sarcocystis canis or a very closely related Sarcocystis sp. in the affected muscle and brain tissues. The main lesion described in previously reported cases of fatal sarcocystosis in bears was necrotizing hepatitis. Fatal encephalitis associated with this parasite represents a novel presentation of sarcocystosis in bears. Sarcocystosis should be considered a differential diagnosis for nonsuppurative encephalitis in bears.     

Experimental evidence for the influence of cumulus-oocyte-complexes on sperm release from the porcine oviductal sperm reservoir

In the pig, a temporal relationship is suggested between sperm release from the sperm reservoir (SR) and ovulation, but the mechanism(s) is still under discussion. In two experiments, the influence of transferred ova on the release of SR-spermatozoa at ovulation and the effect of supplementation with non-sulfated glycosaminoglycan hyaluronan (HA) on embryo development and the number of accessory spermatozoa, respectively, were examined. PMSG/hCG primed ovectomized gilts that had previously received endoscopic low-dose insemination into the cranial uterine horn were used as an experimental model. After salpingectomy, tubal segments (ampulla, cranial, and caudal isthmus) were flushed and sperm numbers or respective accessory spermatozoa were counted. In Experiment 1, the distribution of the sperm population was altered in the presence of cumulus-oocyte-complexes (COCs). A higher proportion of spermatozoa was found after transfer of COCs into one oviduct in the ampulla and cranial isthmus segments compared with the controls (17.5 vs. 4.9%, p<0.05). In Experiment 2, the quality of the transferred ova and treatment influenced the presence of accessory spermatozoa. Transfer of COCs together with HA increased (p<0.05) the number of accessory spermatozoa compared with the other treatment groups and was similar to those in the "undisturbed" controls. No modifications were obtained regarding mean blastomere numbers (2.6 +/- 0.2 to 3.1 +/- 0.2). In summary, this study was demonstrated that cumulus-oocyte-complexes may be involved in triggering sperm release from the pig oviductal SR and that HA might be related to sperm release.     

Anisakiasis with proventricular perforation in a greater shearwater (Puffinus gravis) off the coast of Georgia, United States

A juvenile male greater shearwater (Puffinus gravis) found off the Georgia coast, United States, was lethargic, hypothermic, dehydrated, and emaciated. It was provided with supportive care but subsequently died. The coelomic cavity contained fibrinous fluid intermixed with hundreds of nematodes, some of which protruded through a perforation in the proventriculus. Nematodes were embedded in the wall of the proventriculus from the lamina propria to the serosal surface and were surrounded by granulocytes, epithelioid macrophages, multinucleate giant cells, lymphocytes, and fibroplasia. A full-thickness tear in the proventriculus was lined by heterophils and multinucleate giant cells. The nematode histomorphology was consistent with Anisakis spp. The definitive hosts of Anisakis spp. are marine mammals, which are infected through ingestion of parasitized fish and crustaceans. Marine birds are aberrant hosts of Anisakis spp., and young, inexperienced, immunocompromised birds might be more susceptible to severe infestations. Fatal anisakiasis is rarely reported in birds.     

Pharmacokinetics of imidocarb dipropionate in white-tailed deer (Odocoileus virginianus) after single intramuscular administration

This study was designed to investigate the pharmacokinetics of imidocarb, a carbanilide derivative, in white-tailed deer (Odocoileus virginianus). The pharmacokinetic properties of a single intramuscular (IM) dose of imidocarb were determined in 10 deer. A single IM injection of 3.0 mg/kg imidocarb dipropionate was administered, and blood samples were collected prior to, and up to 48 hr after imidocarb administration. Plasma imidocarb concentrations were determined by high-performance liquid chromatography with ultraviolet detection. The disposition of plasma imidocarb was best characterized by a two-compartment open model. The mean ± SE maximal imidocarb concentration in deer was 880.78 ± 81.12 ng/ml at 38.63 ± 5.30 min postinjection. The distribution phase had a half-life (t_1/2α) of 25.90 ± 10.21 min, and plasma imidocarb concentration declined with a terminal elimination half-life (t_1/2β) of 464.06 ± 104.08 min (7.73 ± 1.73 hr). Apparent volume of distribution based on the terminal phase (V_Z/F) was 9.20 ± 2.70 L/kg, and apparent total body clearance (Cl/F) was 15.97 ± 1.28 ml min⁻¹ kg⁻¹.     

The investigation of interspecies diversity of erythrocyte aggregation properties by two different photometric methods in four animal species

Among the haemorheological parameters, red blood cell (RBC) aggregation shows the largest interspecies diversity, and often controversial data can be found in the literature, besides the methodology‐dependent issues. In this present investigation, we compared four experimental/laboratory animal species’ RBC aggregation by two different photometric methods for better revealing the differences. Blood samples (K3‐EDTA, 1.5 mg/ml) were taken from female animals: 16 inbred mice (Mus musculus, cardiac puncture), 15 outbred rats (Rattus norvegicus, caudal caval vein puncture), 15 beagle dogs (Canis canis, cephalic vein) and 23 juvenile pigs (Sus scrofa domesticus, medial saphenous vein). Haematological parameters (microcell counter) and RBC aggregation (light transmission and syllectometry‐laser backscatter methods) were determined within 2 h after sampling. Describing the first 5–10 s of the aggregation process, additional parameters were calculated out of the syllectometric raw data. Standardized difference was calculated to determine the sensitivity of the two devices. Parameters describing the extent and magnitude of red blood cell aggregation showed the lowest values in the rat and the highest in the pig and canine blood. In turn, parameters describing the kinetics of aggregation showed the lowest values in the mouse and the highest in the rat. The standardized difference values for the laser backscattering method were 2–4 times larger vs. the light transmission one. The magnitude of the differences was not consequent in the aggregation parameters. These comparative results show that the laser backscattering method can detect the RBC aggregation differences between the investigated species more sensitively than the light transmission method.     

Comparison of the nutritional profile of glyphosate-tolerant corn event NK603 with that of conventional corn (Zea mays L.)

The composition of glyphosate-tolerant (Roundup Ready) corn event NK603 was compared with that of conventional corn grown in the United States in 1998 and in the European Union in 1999 to assess compositional equivalence. Grain and forage samples were collected from both replicated and nonreplicated field trials, and compositional analyses were performed to measure proximates, fiber, amino acids, fatty acids, vitamin E, nine minerals, phytic acid, trypsin inhibitor, and secondary metabolites in grain as well as proximates and fiber in forage. Statistical analysis of the data was conducted to assess statistical significance at the p < 0.05 level. The values for all of the biochemical components assessed for corn event NK603 were similar to those of the nontransgenic control or were within the published range observed for nontransgenic commercial corn hybrids. In addition, the compositional profile of Roundup Ready corn event NK603 was compared with that of traditional corn hybrids grown in Europe by calculating a 99% tolerance interval to describe compositional variability in the population of traditional corn varieties in the marketplace. These comparisons, together with the history of the safe use of corn as a common component of animal feed and human food, support the conclusion that Roundup Ready corn event NK603 is compositionally equivalent to, and as safe and nutritious as, conventional corn hybrids grown commercially today.     

Impact of fertilisation practices on nitrogen leaching under irrigation

Field experiments were carried out over a 2-year period on a loamy soil plot under corn in Montpellier (south-east France). The effectiveness of improved irrigation practices in reducing the adverse impact of irrigation on the environment was assessed. Different irrigation and fertiliser treatments were applied to identify the best irrigation and fertilisation strategy for each technique (furrow and sprinkler) to ensure both good yields and lower NO₃^- leaching. No significant differences in corn yield and NO₃^- leaching were found for the climatic scenario of 1999 between sprinkler and furrow irrigation during the irrigation season. Following the rainy events occurring after plant maturity (and the irrigation season), differences in N leaching were observed between the treatments. The study shows that both the fertiliser method, consisting of applying a fertiliser just before ridging the furrows, and the two-dimensional (2D) infiltration process, greatly influence the N distribution in the soil. N distribution seems to have a beneficial impact on both yield and N leaching under heavy irrigation rates during the cropping season. But, under rainy events (particularly those occurring after harvesting), the N, stored in the upper part of the ridge and not previously taken up by plants, can be released into the deeper soil layers in a furrow-irrigated plot. In contrast, the 1D infiltration process occurring during sprinkler irrigation events affects the entire soil surface in the same way. As a result the same irrigation rate would probably increase N leaching under sprinkler irrigation to a greater extent than under furrow-irrigation during an irrigation period. In order to assess the robustness of these interpretations derived from soil N-profile analysis, a modelling approach was used to test the irrigation and fertilisation strategies under heavy irrigation rates such as those occurring at the downstream part of closed-end furrows. The RAIEOPT and STICS models were used to simulate water application depths, crop yield and NO₃^- leaching on three measurement sites located along the central furrow of each treatment. The use of a 2D water- and solute-transport model such as HYDRUS-2D enabled us to strengthen the conclusions derived from the observations made on the N distribution under a cross-section of furrow. This model helped to illustrate the risk of over-estimation of N leaching when using a simplified 1D solute-transport model such as STICS.     

Promotion of tissue inflammation by the immune receptor Tim-3 expressed on innate immune cells

CD4+ T helper 1 (TH1) cells are important mediators of inflammation and are regulated by numerous pathways, including the negative immune receptor Tim-3. We found that Tim-3 is constitutively expressed on cells of the innate immune system in both mice and humans, and that it can synergize with Toll-like receptors. Moreover, an antibody agonist of Tim-3 acted as an adjuvant during induced immune responses, and Tim-3 ligation induced distinct signaling events in T cells and dendritic cells; the latter finding could explain the apparent divergent functions of Tim-3 in these cell types. Thus, by virtue of differential expression on innate versus adaptive immune cells, Tim-3 can either promote or terminate TH1 immunity and may be able to influence a range of inflammatory conditions.