Mutants of rabies viruses in skunks: immune response and pathogenicity.

In studies to develop an oral rabies vaccine for wildlife, the immune response to and pathogenicity of two types of mutants of rabies viruses were examined. Forty-five small plaque mutants were selected from cultures of ERA rabies virus treated with 8-azaguanine or 5-fluorouracil and tested for pathogenicity in mice. Two of these mutants AZA 1 and AZA 2 (low pathogenicity in mice) were given to skunks by oral (bait), intestinal (endoscope) and intramuscular routes. Additionally, challenge virus standard (CVS) rabies virus and mutants of this and ERA rabies virus (CVS 3766 and 3713, and ERA 3629) that were resistant to neutralization by specific antiglycoprotein monoclonal antibodies (and apathogenic in mice) were tested by various routes in skunks. Skunks given AZA 1 and AZA 2 were challenged at three months postinoculation with street rabies virus. After oral administration, there were very low rates of seroconversion with AZA 1 and AZA 2 and on challenge only 2/7 given AZA 1 and 1/8 given AZA 2 survived. None of the skunks given the other mutants orally seroconverted. AZA 2 produced a high rate of seroconversion (8/8) by the intestinal route and all challenged skunks in this group survived (7/7). All skunks vaccinated intramuscularly with AZA 1 (4/4) or AZA 2 (4/4) developed high levels of rabies neutralizing antibodies and survived challenge. The mutant CVS 3766, while apathogenic when given intracerebrally to adult mice, was consistently pathogenic by this route (and intranasally) in skunks. These results demonstrate that skunks are highly resistant to oral immunization by live rabies virus vaccines and that pathogenicity (by intracerebral route) of the mutant CVS 3766 is markedly different in mice and skunks.     

A Quantitative Flurometric Assay for the Measurement of Antibody to Pasteurella haemolytica in Cattle

A rapid, simple fluorometric method is described for measuring antibody to Pasteurella haemolytica in sera of cattle. Various antigen preparations were compared for the test including live, formalin-killed and phenol-killed P. haemolytica. A preparation composed of formalin-killed organisms from a 22 hour culture gave consistent results and was used in the studies. The test was reproduciable with percent coefficients of variation for fluorescent signal unit values on ten or more replicate samples ranging from 5.7 to 28.0. Sera from calves vaccinated by aerosol exposure to live P. haemolytica had up to a five-fold increase in antibody titer as measured by the flurometric method test during a 21 day period. Fluorometric method titers were comparable to those obtained by the indirect bacterial agglutination test. There was no seroconversion to P. haemolytica in calves vaccinated by aerosol exposure of P. multocida. The major advantages of the fluorometric method test over conventional methods are that the assay does not require serial dilutions of serum samples and thus limits time and effort to determine antibody titers.     

Composition, stability, and bioavailability of garlic products used in a clinical trial

In support of a new clinical trial designed to compare the effects of crushed fresh garlic and two types of garlic supplement tablets (enteric-coated dried fresh garlic and dried aged garlic extract) on serum lipids, the three garlic products have been characterized for (a) composition (14 sulfur and 2 non-sulfur compounds), (b) stability of suspected active compounds, and (c) availability of allyl thiosulfinates (mainly allicin) under both simulated gastrointestinal (tablet dissolution) conditions and in vivo. The allyl thiosulfinates of blended fresh garlic were stable for at least 2 years when stored at -80 degrees C. The dissolution release of thiosulfinates from the enteric-coated garlic tablets was found to be >95%. The bioavailability of allyl thiosulfinates from these tablets, measured as breath allyl methyl sulfide, was found to be complete and equivalent to that of crushed fresh garlic. S-Allylcysteine was stable for 12 months at ambient temperature. The stability of the suspected active compounds under the conditions of the study and the bioavailability of allyl thiosulfinates from the dried garlic supplement have validated the use of these preparations for comparison in a clinical trial.     

The role of flow dependency and water availability in fish assemblage homogenization in tributaries of the Chattahoochee River,Alabama, USA

The availability of water is becoming more variable as flow regime alterations intensify both locally and on a global scale. This pattern has been observed in the tributaries of the central Chattahoochee River System as water withdrawal and natural drought have caused flows to diminish. Stream discharge has steadily decreased in the study area by 31% over the last 50 years, and during this time, fish assemblage homogenization has occurred throughout the system. Our primary objective was to assess whether spawning mode and water availability are catalysts for fish assemblage change in tributaries of the Chattahoochee River System. We found that species which prefer or have adapted to low flow conditions (e.g. blacktail shiner Cyprinella venusta, redbreast sunfish Lepomis auritus and blackbanded darter Percina nigrofasciata) are becoming dominant and replacing historically dominant fluvial specialists. When examining short‐term changes between a dry year (2009) and a wet year (2010), some species preferring higher flows such as the highscale shiner Notropis hypsilepis were detected during the wet year, suggesting that the amount of water is important for the presence of these species in upstream reaches where they were historically abundant. Species that can reproduce successfully in low flow conditions are thriving and expanding their native ranges as more habitat becomes suitable. Species that require higher flows for part of their life cycle, however, appear to be declining from upstream areas as water availability continues to decrease, despite temporary recovery from downstream refugia during wet years.     

Duration of immunity in foxes vaccinated orally with ERA vaccine in a bait.

Red foxes (Vulpes vulpes) vaccinated orally with the ERA strain of rabies vaccine in a bait were challenged after 83 mo. Ten of 11 foxes that had seroconverted following vaccination resisted challenge with a virulent rabies virus which produced clinical signs of rabies in 6 of 6 unvaccinated foxes. Five of 11 vaccinated animals retained titers of rabies virus neutralizing antibody throughout the period. Although 6 of 11 had no detectable antibody at the time of challenge, 5 of these 6 resisted challenge and had an anamnestic response, as indicated by elevated titers of antibody when measured at day 77 postchallenge. These results show that foxes can be immunized successfully with a single oral dose of ERA vaccine, probably with protection against a lethal rabies challenge, for at least 7 y.     

Demographic survey of veterinarians employed in western Canada

The objective of this study was to generate demographic data on veterinarians working in western Canada. A sample of 551 veterinarians was randomly selected from a population of 2474 veterinarians employed in western Canada, 425 (77.1%) of whom responded to the survey. The respondents were evenly split between males (53.1%) and females (46.9%). More than half (58.0%) of the private practitioners practised exclusively on companion animals (small animals and horses), while 2.9% devoted 100% of their time to food animals. There were 351 respondents who had had ≥ 2 employers since graduation; 80% of those who had begun their careers in companion animal (CA) practice had remained in this type of practice, while 54.3% of those who had begun their careers in mixed animal practice had switched to CA practice. Analyses of wage and workload data from 85 full-time veterinary employees showed that CA practitioners worked the fewest hours/week (47.0), had the least number of evenings on-call/month (3.7), and earned the highest hourly wage ($35.79) as compared with non-CA practitioners.     

Crop loss and control of Pseudocercospora fruit and leaf spot of citrus in Ghana

Pseudocercospora fruit and leaf spot (PFLS) of citrus, caused by Pseudocercospora angolensis, was recently described in Ghana and has spread in most citrus-growing areas of the country. A survey of PFLS incidence was conducted in the Eastern Region. Orchards were georeferenced and data on altitude, annual mean temperature, and annual precipitation were obtained from the WorldClim database. Fruit drop due to PFLS and other pests and diseases was evaluated in three orchards. Field efficacies of 4-week, 6-week and 8-week schedules with carbendazim + mancozeb were evaluated in the major and minor fruit production seasons. Ordinal logistic regression and generalized linear models were fitted in each case according to the nature of the data and possible overdispersion. Disease incidence in the sweet orange orchards surveyed was 25–100 %, with higher values in higher altitude areas exposed to lower temperatures and higher rainfall. PFLS was the main problem causing yield losses, associated with 84–87 % of fruit dropped on the orchard floor. PFLS severity on shoots and incidence on fruit 12 weeks after full bloom was significantly reduced by all fungicide schedules evaluated. The effects of fungicides on PFLS severity in fruit at harvest were not substantial because of intense fruit drop. The 8-week schedule showed the lowest effectiveness in reducing fruit drop and thus the 6-week schedule is preferred. Further research is needed for an integrated management of PFLS in Ghana.