Immunity of specific pathogen-free lambs to challenge with an aerosol of Pasteurella haemolytica biotype A serotype 2. Pulmonary antibody and cell responses to primary and secondary infections

Specific pathogen-free (SPF) lambs previously exposed to an aerosol of P. haemolytica biotype A serotype 2 (A2) were immune to subsequent challenge with an aerosol of P. haemolytica A2. Untreated control lambs were not immune to this challenge. The local immune responses of the lung to these challenges were examined. High IgG and IgA titres to P. haemolytica and high levels of opsonizing antibody against P. haemolytica were present in the lung washings from previously infected immune lambs at autopsy, seven days after the second infection. Lung washings from control lambs, 7 days after challenge with P13 virus and P. haemolytica A2, had no IgG titres, very little opsonizing activity but did have IgA titres which were significantly higher than in unchallenged control lambs. The cellular response of animals challenged with P13 virus and P. haemolytica was significantly greater than that of unchallenged controls or of lambs exposed only to P. haemolytica. However, this finding was complicated by the response to P13 virus. Lymphocytes from lung washings of all lambs failed to respond in a lymphocyte stimulation test to phytohaemagglutinin while blood lymphocytes did respond. There was little specific response to P. haemolytica antigen in the test.     

Blood coagulation parameters in clinically normal intensively farmed red deer (Cervus elaphus) and fallow deer (Dama dama)

Reference ranges for five parameters of blood coagulation were established in clinically normal farmed fallow deer (Dama dama) and red deer (Cervus elaphus). Storage of plasma at -15 degrees C resulted in small increases in activated partial thromboplastin time (APTT) for both species, and in the one stage prothrombin time (OSPT) of fallow deer, between days 1 and 30. These times were then stable between days 30 and 60. The fibrinogen levels of fallow deer plasma showed a small apparent increase over 60 days. These storage artefacts were not of sufficient magnitude to preclude the use of such plasma for diagnostic purposes although they could limit its use in research. Levels of fibrin/fibrinogen degradation products (FDP) were not affected by storage at -15 degrees C. Rabbit brain thromboplastin appeared adequate for OSPT determination in both species. The activated clotting time (ACT) is recommended as a field test for screening the intrinsic and common pathways of blood coagulation in deer.     

Endotoxin from Fusobacterium necrophorum of bovine hepatic abscess origin.

The endotoxic activity of Fusobacterium necrophorum bov 5 was investigated. The supernatant (S) fluid and cell wall (CW) preparation, obtained after differential centrifugation of the ruptured cell mass, were lethal for mice. The toxicity of the S fluid was stable during prolonged storage, treatment with formalin, and heating for 15 minutes at 80, 100, and 121 C, but was destroyed by alkaline hydrolysis with 0.25 N NaOH. The toxic factor was found in a high molecular weight (MW) fraction after gel filtration. The properties exhibited by the toxic S fluid resembled those of endotoxic lipopolysaccharide (LPS). Extracted and partially purified LPS (endotoxin) from F necrophorum bov 5 demonstrated a mouse median lethal dose (mouse LD50) of 16.8 mg/kg of body weight. The toxic LPS material, a high molecular weight moiety as estimated by gel filtration, was resistant to ribonuclease (RNase), deoxyribonuclease (DNase), and pronase treatment. A positive Shwartzman reaction (median skin lesion dose (SLD50) equal to 3.32 mug/kg of body weight) and biphasic fever response (minimal dose required to produce a fever index of 40 sq cm which falls on the linear portion of dose-response curve (FL40) equal to 0.41 mug/kg of body weight) further indicated the toxin was endotoxin in nature. The LPS from F necrophorum bov 5 was less toxic than Salmonella typhimurium LPS; but had considerable toxicity for experimental animals. The toxic activity of the partially purified F necrophorum bov 5 endotoxin was separated into 2 fraction regions by diethylaminoethyl (DEAE)-cellulose chromatography. The data provide evidence for the production of a potent endotoxin, possibly composed of more than one toxic component, which may be released upon cell disruption.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

Development of a model animal welfare act curriculum

Animal-welfare issues are often controversial and frequently have an emotional component. Veterinarians have extensive knowledge, experience, and scientific perspective and are arguably the professionals best suited to advise and develop recommendations on animal welfare. The development of an Animal Welfare Act (AWA) teaching module is a first step toward educating veterinary students about animal welfare. This article presents the current development status of this curriculum project, which is intended to be a valuable addition to the evolving veterinary education on animal welfare.     

Effects of chemical and heat treatments on ethylene production in soil

Factors influencing C₂H₄ production in a silt loam were investigated in an effort to determine the source of this gas in soil. Air-dried samples of soil in glass vials were moistened to about ?10 kPa, sealed with rubber septa, and incubated at 30 or 35°C with an original atmosphere of air or O₂-free N₂. C₂H₄ concentrations in the vials were determined by gas chromatography.Addition of the antibacterial agents chloramphenicol or novobiocin to the soil inhibited C₂H₄ production, whereas the antifungal agent cycloheximide had no effect. Sodium azide and sodium cyanide also reduced C₂H₄ production. Treatment of the soil with moist heat (i.e. passing a steam-air mixture through it) at 80°C for 30 min failed to reduce the ability of the soil to produce C₂H₄ during subsequent incubation at 30°C, but autoclaving it twice at 121°C prevented C₂H₄ production. As with nonheated soil, C₂H₄ production from soil treated at 80°C was prevented by novobiocin but not by cycloheximide. Only about 10% of the bacteria isolated from nontreated soil were spore-formers. In contrast, 95–98% and possibly more of the bacteria isolated from heat-treated soil were spore-formers, including those in soil which was heat-treated and then incubated moist at 30°C for an additional 3 days before dilution plating. Addition of methionine had no effect on the production of C₂H₄ in anaerobic soil, whereas ethionine, chlorogenic acid, and ethylenediaminetetraacetic acid (EDTA) all enhanced C₂H₄ production. Ethionine, but not chlorogenic acid or EDTA, also resulted in considerable C₂H₄ accumulation in autoclaved soil; the C₂H₄ detected in ethionine-amended soil was apparently nonmicrobial in origin. Soil samples incubated at constant temperatures of 30, 50, or 70°C all produced C₂H₄.The results collectively indicate that C₂H₄ in soil is most likely produced by facultative or strictly anaerobic bacteria which are probably spore-formers and may also be thermophilic. Several isolates of spore-forming bacteria were inoculated into autoclaved soil, but none produced appreciable amounts of C₂H₄ under the test conditions.     

Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.