A farm investigation into swayback in a herd of goats and the result of administration of copper needles

Clinical, biochemical and pathological findings led to a diagnosis of swayback in a herd of goats. Trace element values in soil, pasture and supplementary feed were measured, and copper oxide needles administered to the adult goats. This form of treatment had a more limited effect in these goats than is reported in sheep.     

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infective Teladorsagia circumcincta in the abomasum of sheep

In the present study we have employed an in vitro organ challenge model to study the post-challenge responses in parasite na?ve and immune gastric tissue of sheep, in an attempt to identify the host derived factors involved in immune exclusion of Teladorsagia circumcincta larvae. Proteins present in the epithelial cells and mucus from ovine abomasa following parasite challenge in previously na?ve and immune animals were analysed through Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-Tof)-MS and shotgun proteomics. MALDI-ToF analysis of epithelial cell lysates revealed that a number of proteins identified were differentially expressed in na?ve and immune cells. These included intelectin and lysozymes, which were present at higher levels in epithelial cell lysates derived from immune samples. A large number of proteins were identified in the mucosal wash from immune tissue which were not present in the mucosal wash of the na?ve tissue. Some of these proteins were present in washes of immune tissue prior to the parasite challenge including immunoglobulin A, galectin 14 and 15 and sheep mast cell protease 1. However, other proteins, such as calcium activated chloride channel and intelectin were only detected in the washings from the challenged tissue. The latter may be related to an enhanced mucus release, which may result in entrapment of infective larvae and thus reduced establishment in tissue that has been previously challenged with the parasite. In conclusion, several proteins have been identified which may be involved, either directly or indirectly, in the exclusion and immune elimination of incoming infective larvae. In the present study, the usefulness of the in vitro model has been confirmed, and the global proteomic approach has identified proteins that had not previously been associated with parasite exclusion from abomasal mucosa, such as the calcium activated chloride channel.     

Analysis of DNA relationships among Aeromonas species by RAPD (randomly amplified polymorphic DNA) typing

Genetic differences between a collection of aeromonads were studied in two laboratories by analysis of randomly amplified polymorphic DNA (RAPD). A single randomly designed primer, generated reproducible profiles of genomic DNA in both laboratories for Aeromonas salmonicida subspecies salmonicida, although the profiles differed between laboratories. Analysis of atypical strains of A. salmonicida and isolates of the A. hydrophila group produced scattered profiles in both laboratories. The uniform fingerprints produced for A. salmonicida subspecies salmonicida indicate genomic homogeneity. The scattered RAPD profiles of the motile aeromonads demonstrate the genomic diversity of this group. A group of unspeciated motile aeromonads gave uniform fingerprints, suggesting the possibility of a genomically homogeneous species. Although the RAPD technique is susceptible to the effects of minor technical variations, this study has demonstrated that where there is DNA similarity, it can be recognized, and where there is diversity, differentiation can be made. RAPD promises to be useful in epidemiological studies for rapid identification of bacteria where a source of reference DNA is available and may be useful in preliminary investigations of relatedness within groups.     

Persistence of Complex Virulences in Populations of Phytophthora infestans in Western Washington

ABSTRACT Isolates of Phytophthora infestans, collected from bittersweet, hairy nightshade, petunia, potato, potato vine, and tomato in western Washington, 1998 to 1999, were evaluated for virulence complexity as well as mating type, metalaxyl insensitivity, allozymes of glucose-6-phosphate isomerase and peptidase, and DNA fingerprint with the RG57 probe. Results were compared with those from similar collections made in the same region during the 1990s. Generally, virulence complexity was high for most of the isolates regardless of year, genotype, or host. No marked shift in virulence complexity was evident for the populations studied, and unnecessary virulences were maintained. During 1998 and 1999, isolates of the US-8 and US-11 genotypes had 4 or more virulence factors. US-8 isolates averaged 8.2 and 9.3, whereas US-11 isolates averaged 5.4 and 6.3 virulence factors. The frequency of US-8 isolates that were sensitive to metalaxyl ranged from 5% in 1998 to 72% in 1999. All of the US-11 isolates tested in 1998 and 1999 were insensitive to metalaxyl. From 1996 to 1999 on potato, the recovery of US-8 increased, whereas the recovery of US-11 decreased. No evidence of new genotypes or sexual recombination was found. Western Washington was a desirable location for screening germ plasm for durable resistance to late blight due to the high frequency and persistence of complex virulences.     

Characterization of the in vitro core surface proteome of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host–pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host–pathogen interactions.     

Survival of Escherichia coli in the intestine of fish

Establishment and persistence of Escherichia coli in the intestine of rainbow trout. Oncorhynchus mykiss (Walbaum), was investigated. Infection was achieved by ingestion of contaminated feed but not by bath exposure. At 15°C. E. coli was found to increase in number in the intestine offish after an initial decline, and could still be detected after 4 days. At 6°C it was detected for 2 days but the numbers declined steadily. A similar trend was observed when extracted gut content was inoculated with E. coli in vitro; after an initial decline, bacterial growth recovered. Gut content in vitro also was a suitable environment for plasmid-mediated transfer of resistance. Oxytetracycline resistance was transferred successfully from Aeromonas salmoniclda to E. coli. This could not be reproduced in vivo, perhaps because the inoculum of A. salmoniclda achieved per fish was not high enough as it did not withstand drying well on the feed pellets. Introduction by gavage was also tried but failed to establish an A. salmonicida infection. These results show that at temperatures around 15°C, the presence of E. coli in fish need not be an indicator of recent passage through polluted waters. It may be a consequence of infection established many days before, and perhaps some distance away. Furthermore, although transfer of antibiotic resistance from a fish pathogen did take place in the environment of fish intestinal content, the likelihood of this being a high-risk route for acquisition of resistance by bacteria that can affect humans is low.     

Antibiotic protection against recrudescence of latent Aeromonas salmonicida during furunculosis vaccination

Problems of temporary immunosuppression following vaccination against Aeromonas salmonicida infection had to be overcome in the development of a furunculosis vaccine. Empirical observations have indicated that immunosuppression persists for some time after vaccination, rendering fish, especially subclinical carriers of A. salmonicida, highly vulnerable to bacterial invasion. The efficacy of simultaneous application of furunculosis vaccine and a long-lasting amoxycillin preparation to a population of Atlantic salmon smolts was evaluated. Control groups were treated with either vaccine alone, amoxycillin alone or were untreated. Moderate stress, simulating smolt transfer with a 5°C temperature rise, resulted in a rapid outbreak in mortalities reaching 100% in the vaccinates. Losses among the untreated controls were more gradual and rose to about 50%. Both amoxycillin-treated groups survived well. Further severe stress resulted in total mortalities among the untreated fish but no further losses in the amoxycillin groups. Four months after vaccination, evidence of a specific immune response was confirmed by ELISA, demonstrating circulating antibodies in the blood of vaccinates. In a severe and in a moderate challenge with A. salmonicida., the relative specific protection was 63 and 86%, respectively. Thus, effective protection against furunculosis was achieved without jeopardizing the stock during the vaccination process and with elimination of the carrier state.     

Acute phase response in horses: changes in plasma cation concentrations after localised tissue injury

An acute phase reaction was elicited in four horses to which Freund's adjuvant was administered intramuscularly. The localised inflammation was accompanied by changes in the plasma concentrations of copper, iron and zinc. The plasma copper concentration, the plasma ceruloplasmin copper concentration and the ceruloplasmin oxidase activity in the plasma steadily increased to a maximum 24 days after the administration of the adjuvant. At this time, the plasma copper concentration was 2.2 micrograms/ml, a 90 per cent increase over the baseline concentration. The ratio of the concentration of plasma ceruloplasmin copper to plasma copper remained constant, indicating that the non-ceruloplasmin bound copper component of the plasma is also an acute phase reactant in the horse. The plasma zinc and iron concentrations decreased to 59 per cent and 30 per cent of their respective baseline concentrations and the severity of the inflammation appeared to influence the plasma concentrations of each metal. Weak correlations between the plasma fibrinogen concentration and the plasma copper and zinc concentrations of 25 horses with plasma fibrinogen concentrations of 5 g/litre or greater indicated that a single measurement of plasma copper concentration is not useful in the diagnosis of non-specific inflammatory disorders of the horse. However, the results suggest that the plasma copper concentrations in serial samples may be used to monitor the resolution of inflammatory disorders in the horse.     

Opposing offspring sex ratio variations with increasing age and weight in mouflon mothers (Ovis musimon).

There are two main theories explaining offspring sex biases in polygynous mammals. Trivers and Willard (1973) argue that mothers with greater reproductive resources should invest in the sex with the greater variance in reproductive success, usually sons. In contrast, because daughters in many polygynous mammals stay with their mother and compete with her for food, Local Resource Competition theory (e.g. Clark, 1978; Silk, 1983) predicts that the mothers with the greatest reproductive resources should invest in daughters. We investigated the strategy of sex allocation of a captive, outdoor population of 139 mouflon mothers, Ovis musimon, kept in a game state. A complex picture emerged in which, despite weight and body condition being correlated with age in female mouflons, mothers lambed more daughters with increasing age but also, within a given age, gave birth to more sons with increasing weight. Results may be useful in game management aimed at increasing the recruitment or quality of males in managed populations.