Characterization of BoHV-1 gE envelope glycoprotein mimotopes obtained by phage display

A phage-displayed peptide library was screened using four mAbs directed against bovine herpesvirus 1 (BoHV-1) gE glycoprotein to identify peptides mimicking this glycoprotein. The selected mimotopes allowed us to characterize the epitopes corresponding to the mAbs as continuous and proteinic and to consider using these peptides in further studies. One epitope has been clearly located at the C-terminus of the protein (amino-acids 561-569). The three other mAbs enabled us to stress the immunogenic relevance of the proline-rich motifs of gE. Selected peptides showed no clear sequence identity with gE, but there is a clear link between gE proline-rich regions and the amino-acid composition of the mimotopes. The proline-rich motifs of gE are potentially located in flanking regions involved in the gE/gl glycoprotein complex formation. N-terminal fusion to pill or pVIII filamentous phage protein, C-terminal fusion to the T7 phage capsid protein, biotinylated synthetic peptides and insertion between the non-cleaved CX leader sequence and the C-terminal part of Caulobacter crescentus RsaA protein have been tested in order to increase the valency of a model peptide. We have diverted the C. crescentus expression system and proven its usefulness using the RsaA protein as a scaffold displaying the peptides of interest. Comparison between these different display systems in an indirect ELISA, indicates that the C. crescentus expression and the T7 phage display systems have some major advantages.     

CliniTox: the computer-based information system for poisoning in farm animals

Poisonings can cause reduced performance and severe economic loss in farm animals. A swift and targeted action is required from veterinarians. We have established a computer-based decision support system for poisonings in ruminants and pigs. The system offers access to the most recent information available in clinical toxicology of farm animals. Towards this goal, relevant reports from the peer-reviewed literature were evaluated and organised according to the requirements of a structured database. The information provided for each toxic substance includes a summary of its chemical and physical properties, sources, commercial uses or natural occurrences, toxicokinetic data, mechanisms of action, threshold doses, clinical symptoms with brief case reports, sampling and analytical results, post-mortem abnormalities, differential diagnoses, therapeutic guidelines and references to the literature. This decision support system has been programmed with two user-friendly search functions: a search tool that allows the choice of clinical and pathological findings, and another function that serves to find a substance using its chemical name, the class of compounds to wich it belongs, a possible source or one of its main applications. CliniTox can be accessed directly via our webserver (http://www.clinitox.ch).     

Effect of the seaweed Ascophyllum nodosum on lambs during forced walking and transport

The objective of this study was to determine the effect of feeding Ascophyllum nodosum (ANOD) to lambs at 0 (control), 0.5, 1, or 2% of DMI/d for 2 wk on lamb physiology in response to forced walking and transport during hot weather. Forty-four lambs (26 kg +/- 4.3) were used, and each lamb swallowed 3 gelatin capsules filled with ANOD or their normal 16% CP, pelleted grain ration twice daily, with the amount of ANOD dependent on the treatment. The amount of ANOD did not affect ear canal temperature or cortisol concentrations during 60 min of forced walking. The range between the minimum and maximum ear canal temperature for each lamb during 12 h of transport was narrower in lambs receiving the 2% ANOD than the control group (P = 0.05), and the 2% ANOD group also had lower (P = 0.05) ear canal temperatures than the control group during hot periods of transport. After 4 (P = 0.09) and 8 h (P = 0.05) of transport, the control group tended to have greater cortisol concentrations than the 2% ANOD group. Many differences among treatments were found in plasma protein, albumin, calcium, phosphorus, glucose, blood urea nitrogen, aspartate aminotransferase, magnesium, sodium, potassium, and chloride concentrations posttransport; mainly, the control and 0.5% ANOD groups had greater (P < 0.05) concentrations than the other 2 treatments. Aldosterone concentrations were greater in the control and 0.5% ANOD group than in the 1 and 2% ANOD groups before transport, whereas the concentrations were not different after transport, suggesting pretransport concentrations were lowered by supplementation. The 1 and 2% ANOD groups lost more BW than the control group as a result of transport (P = 0.04). After transport, no differences were observed in the latency for lambs to drink, eat, or lay. There was a suppression of the IgG and IgM antibody responses at 4 and 7 d after administration of ovalbumin, with greater ANOD supplementation rates suppressing antibody response the greatest. Although ANOD decreased ear canal temperature in hot periods of transport, stabilized electrolyte concentrations, and decreased cortisol throughout transport, it also suppressed the antibody response indicating that the effect of ANOD on immune function merits further investigation.     

Spontaneous bilateral coxofemoral luxation in four dogs

Spontaneous bilateral coxofemoral luxations and their clinical managements are described in four dogs. Luxations in each case occurred while the patient was in a controlled environment with no possibility of trauma. While all cases had bilateral luxations, the luxations occurred in a staged fashion in two of the four cases. Treatment varied with each case, but included closed reduction, triple pelvic osteotomy, prosthetic capsulorrhaphy, toggle pin, ischioilial pinning, total hip replacement, and femoral head and neck ostectomy. Other than femoral head and neck ostectomy, results were uniformly poor, with reluxation being the most common complication. Evidence of pre-existing degenerative changes was minimal, particularly considering the ages of the patients (range, three to 12 yrs). Histopathological evaluation of joint capsules, round ligaments, and femoral heads are warranted in future cases.     

Technical note: recombinant LHRH fusion protein suppresses estrus in heifers

A recombinant luteinizing hormone-releasing hormone (LHRH) fusion protein was evaluated for its effectiveness in suppression of estrus in heifers. Eight heifers were randomly assigned to two equal treatment groups. Treatments consisted of recombinant ovalbumin-LHRH-7 or recombinant ovalbumin (control). This recombinant chimeric fusion protein consisted of ovalbumin with seven LHRH peptides (ovalbumin-LHRH-7). The plasmid for this protein was expressed in E. coli and was collected and purified as an insoluble protein. One milligram of the respective proteins was suspended in 2 mL of Z-Max adjuvant and administered by intramammary injection three times at 7-wk intervals. Luteinizing hormone-releasing hormone antibody binding was elevated in heifers treated with ovalbumin-LHRH-7 compared to ovalbumin-treated heifers (P < .05). Serum progesterone concentrations (< 1 ng/mL) indicate that the estrous cycle of the four heifers treated with ovalbumin-LHRH-7 was suppressed for a time period ranging from 60 to 238 d, which was different from control heifers (P < .01). Serum progesterone for the control heifers continued to exhibit cyclic profiles over the experimental period. This preliminary study in heifers demonstrated that a chimeric LHRH fusion protein induced elevated concentrations of circulating LHRH antibodies that suppressed estrus for an average of 122 +/- 41 d.     

Digesta kinetics, energy intake, grazing behavior, and body temperature of grazing beef cattle differing in adaptation to heat

The objective of this study was to determine whether digesta kinetics, energy intake (EI, kcal ME intake x kg(-.75) x d(-1)), grazing behavior, or body temperature differed by breed, lactational state, or season of the year among cattle presumed to vary in adaptability to the subtropics. Two-year-old lactating and nonlactating Brahman x Angus (BA; n = 5, n = 5), Tuli x Angus (TA; n = 5, n = 4), and Angus (A; n = 4, n = 4) cows were used. During both early (ES) and late summer (LS), lactating cattle vs nonlactating cattle had greater gastrointestinal tract load (CM2) and EI (P < .01), although passage rate did not differ (P > .48). During LS, lactating cattle had decreased early morning rectal temperatures (P < .05) and spent more time grazing during the day compared with nonlactating cattle (P < .001). Among breeds, A had the largest CM2 (P < .01 compared with BA and P = .068 compared with TA) and accumulated the greatest heat during the day (P < .05). Due to greater daytime shading (P < .01) and less daytime grazing (P < .05), A had lower (P < .05) early morning and comparable (P > .26) late afternoon rectal temperatures compared with BA and TA. With data pooled over both grazing trials, BA cattle had the smallest CM2 (P < .01), and in ES they spent the least amount of time in the shade (P < .001). The TA spent more time in the shade than did BA (P < .001) during ES and less during LS (P < .001) and had similar (P > .28) early morning rectal temperatures compared with BA during ES and LS. During LS, TA spent more time in the sun and less time in the shade than did either A or BA (P < .001). During ES, EI did not differ among breeds (P > .50). During LS, EI for lactating A was greater than for BA and TA (P < .05), and EI for nonlactating BA was less than for A and TA (P < .05). Bite rate per minute for lactating cattle during ES was reduced (P < .03) by increased body condition score. Tuli x Angus cattle appear to be comparable to BA with respect to heat adaptation. It appears that EI demands are greater in a hot environment.     

Nutritional evaluation of sunflower seed and products derived from them. Effect of oil extraction

1. Apparent MEn and oil digestibility of hulled sunflower seed (SFS) and the products derived from the oil extraction process: press extracted SFS (PESFS), sunflower seed meal (SFSM), and press and solvent oils (PO and SO) were determined with 198 cockerels. Recombined products (mix of meal and oil) were also evaluated to study the effect of the 2 consecutive oil extraction steps. Each foodstuff was included in a basal diet, according to the proportions resulting from processing, at 100, 200, and 300 g SFS/kg. 2. Dietary energy value and digestible fat content were linearly related to rate of inclusion of test ingredients. Extrapolation values for AMEn (MJ/kg DM) were: SFS, 16.20; PESFS, 9.46; SFSM, 7.62. A decreasing quadratic trend was also found in the AMEn of PESFS, with interpolation values ranging from 11.77 to 9.33 MJ/kg DM between 70 and 210 g/kg DM of inclusion. No differences were observed between PO and SO. The AMEn of of sunflower oil, calculated from its digestibility, was 33.70 MJ/kg DM. 3. The oil extraction process affected the nutritional value of sunflower products. Recombined materials showed greater values than original foodstuffs: R-SFS, 17.47; R-PESFS, 11.49 MJ/kg DM. The increase in oil digestibility (from 0.814 to 0.862 g/kg DM in SFS; from 0.778 to 0.892 in PESFS) accounted for most of the increase observed in AMEn values. 4. As the form in which oil is incorporated in diets (released or within SFS or PESFS) affects the utilisation of sunflower products, their nutritional value is less than maximal and should not be calculated from their ingredients. Solvent oil seems to be the less available fraction of sunflower oil within SFS as the effect of the 2nd extraction proved to be greater than that of the previous press extraction.     

Synovial fluid analysis in cattle: a review of 130 cases

OBJECTIVE: To compare synovial fluid characteristics of cattle with infectious and noninfectious arthritis. STUDY DESIGN: Retrospective cohort study. ANIMAL OR SAMPLE POPULATION: 130 cattle. METHODS: Synovial fluid was analyzed for total nucleated cell count (NCC), absolute number and percentages of polymorphonuclear (PMN) and mononuclear cells, total protein (TP) concentration, and specific gravity. Cattle were categorized as having infectious or noninfectious arthritis based on physical and lameness examinations, joint radiographs, and microbial culture results. Kruskal-Wallis 1-way analysis of variance was used to compare synovial fluid analysis data from different categories. Selection of cut-off values for the calculation of likelihood ratios, sensitivity, specificity, and positive and negative predictive values was based on examination of the distribution of the data using histograms. RESULTS: Cattle with infectious arthritis had significantly higher numbers of total NNC, PMN cells, TP concentration, and specific gravity (P = .0001) and a significantly higher percentage of PMN cells compared with cattle with noninfectious arthritis (P = .0001). The percentage of mononuclear cells was significantly higher in cattle with noninfectious arthritis (P = .0001). CONCLUSIONS: Synovial fluid analysis is useful for differentiation of infectious and noninfectious causes of joint disease in cattle. CLINICAL RELEVANCE: Cattle with a synovial fluid total NCC > 25,000 cells/microL, a PMN cell count > 20,000 cells/microL or more than 80% PMN cells, and TP > 4.5 g/dL should be considered to have infectious arthritis.     

Infection of a canine macrophage cell line with leishmania infantum: determination of nitric oxide production and anti-leishmanial activity

We have previously shown that resistance to Leishmania infantum in dogs is associated with a Th1 type of immune response. In this study, we use a canine macrophage cell line (030-D) that can readily be infected with this protozoan parasite. Our aim is to further characterize the effector mechanisms involved in killing of Leishmania parasite in dogs. We observed that activation of 030-D cells by incubation with a supernatant derived from a Leishmania-specific T cell line containing IFN-gamma, TNF-alpha and interleukin-2 (IL-2) resulted in enhanced nitric oxide (NO) production by these cells. In addition, we observed enhanced anti-leishmanial activity of infected 030-cells after activation. Both, NO production and anti-leishmanial activity were abrogated by addition of L-N(G)-nitroargininemethyl ester (L-NAME), an analogue of L-arginine. Thus, NO play an important role in the anti-leishmanial activity of these canine macrophages. We propose the infection of the 030-D cell line as a good in vitro model to further investigate parasite-host cell interactions in dogs, a natural host of Leishmania parasites.     

Genetic analysis of vesicular stomatitis virus-New Jersey from the 1995 outbreak in the western United States

OBJECTIVE: To compare molecular associations between the vesicular stomatitis virus (VSV)-New Jersey isolates of the 1995 outbreak with those from previous outbreaks between 1982 and 1985 in the western United States. SAMPLE POPULATION: 23 virus isolates considered representative of the 1995 outbreak of vesicular stomatitis. PROCEDURE: Viral gene coding for surface-envelope protein G was evaluated by use of nucleotide sequencing and phylogenetic analysis. RESULTS: Changes in up to 0.77% of the nucleotide bases and 1.35% of the amino acids were detected among the 1995 viral isolates, whereas changes in up to 3.2 and 2.9% of the nucleotides and amino acids, respectively, were found, compared with the 1982 to 1985 viruses. Insertions or deletions were not found in the entire gene, which spanned 1,554 nucleotide bases. CONCLUSIONS AND CLINICAL RELEVANCE: Phylogenetic analysis indicated that the 1995 VSV-New Jersey belongs to a lineage distinct from that of the 1982 to 1985 viruses that caused previous outbreaks in the western United States. Furthermore, it also is distinct from strains from Central America and from the Georgian Hazelhurst strain.