Molecular and immunological characterisation of Theileria parva stocks which are components of the 'Muguga cocktail' used for vaccination against East Coast fever in cattle

The 'Muguga cocktail' which is composed of three Theileria parva stocks Muguga, Kiambu 5 and Serengeti-transformed has been used extensively for live vaccination against East Coast fever in cattle in eastern, central and southern Africa. Herein we describe the molecular characterisation of the T. parva vaccine stocks using three techniques, an indirect fluorescent antibody test with a panel of anti-schizont monoclonal antibodies (MAb), Southern blotting with four T. parva repetitive DNA probes and polymerase chain reaction (PCR)-based assays detecting polymorphism within four single copy loci encoding antigen genes. The Muguga and Serengeti-transformed stocks exhibited no obvious differences in their reactivity with the panel of MAbs, whereas Kiambu 5 differed with several MAbs. Kiambu 5 DNA was very distinct from the Muguga and Serengeti-transformed isolates in the hybridisation pattern with all four nucleic acid probes, whereas Muguga and Serengeti-transformed isolates exhibited minor differences and could not be discriminated with one of the probes. PCR amplification in combination with restriction fragment length polymorphism analysis indicated that Kiambu 5 was also markedly divergent from the Muguga and Serengeti-transformed stocks within two of the four antigen coding genes. The T. parva Serengeti-transformed stock did not contain a 130 base pair insert within the p67 sporozoite antigen gene, which has been observed previously in most T. parva parasites isolated from buffalo, and could not be discriminated from T. parva Muguga at any of the four single copy loci. Collectively the data indicate that two of the cocktail components T. parva Serengeti-transformed and Muguga are genetically closely related, while the third component Kiambu 5 is quite distinct. Based on the findings, there may be a need to include only one of the T. parva Muguga and Serengeti-transformed components in the immunising cocktail. The study demonstrates the value of molecular characterisation data for monitoring of live vaccines.     

Evaluation of cinnamamide as an avian repellent: Determination of a dose—response curve

The dose-response curve was quantified for the avian repellency of cinnamamide, a derivative of the plant secondary compound cinnamic acid, using a three-day, ‘short-term no-choice’ test. This test was designed to resemble more closely the availability of alternative food in the birds' natural environment than in previous no-choice studies. Cinnamamide reduced food consumption by feral pigeons (Columba livia Gmelin) at concentrations as low as 6.17 μmole g^?1 food (0.09% w/w) and this reduction increased with concentration. An R₅₀ index, the concentration at which food consumption was reduced by 50% was interpolated from the dose-response curve as 18.4 (±0.38) μmole g^?1 food 10.26% w/w). This index can now be matched with those of related compounds in future structure-activity studies. The birds' response to cinnamamide changed over the three-day trial with respect to concentration. Repellency declined at concentrations < R₅₀ and increased at concentrations > R₅₀. This suggests that pigeons become habituated to cinnamamide at low concentrations but at high concentrations they may develop a conditioned aversion. This study provides quantitative confirmation of previous laboratory and field demonstrations of cinnamamide's effectiveness as a non-lethal chemical repellent suitable for reducing avian pest damage.     

How lambs control infection with Ostertagia circumcincta

Two of the most important questions for immunologists studying nematode infections are what effect does the host response have on the parasite and which components of the host response are responsible for these effects. The number of nematodes and the mean length of adult female Ostertagia circumcincta was measured in over 500 6-7 month old lambs of the Scottish Blackface breed. Quantitative genetic analyses indicated that there was substantial genetic variation among lambs in faecal egg counts and in worm length but the analyses provided no evidence for genetic variation in worm numbers. Separate analyses have shown a strong relationship between decreased worm length and decreased worm fecundity. Therefore, genetic resistance in lambs appears to be due to control of worm growth and not to control of worm numbers. The only immune response that is consistently associated with reduced worm length is the IgA response to fourth-stage larvae. The association is remarkably strong. After allowing for the influence of worm number on worm length (density-dependence) essentially all of the variation among deliberately infected lambs in worm length can be accounted for, in a statistical sense, by variation in the strength and specificity of the local IgA response. Therefore, the IgA mediated suppression of worm growth and fecundity appears to be the major mechanism of resistance to O. circumcincta in lambs.     

Modeling of host genetics and resistance to infectious diseases: understanding and controlling nematode infections

This paper considers approaches to modeling the dynamics of infectious disease and the application of such models to nematode parasite infections in ruminants. Particularly, these models are developed to account for host genetics and may be used to assess the effects of using genetics to control nematode infections. Three main issues are critically examined: the infection transmission cycle from pasture to host to pasture, the expected genetic relationships between resistance and performance, and the risks of parasite evolution in response to genetic changes in the host. To obtain answers that are realistic and of practical use, the modeling approaches require a solid grounding in biology. This biology is formalized and described using mathematical techniques, with the models parameterized using experimental or field data. Transmission dynamics have been quantified by modeling and are backed by strong experimental data. Selection for resistance will be successful in reducing egg output, pasture larval contamination and hence subsequent larval challenge. Modeling frameworks have been developed to predict genetic relationships between resistance to infectious disease and performance in general, and genetic correlations predicted for nematode resistance are close to mean published values. These predicted correlations strengthen as the larval challenge increases and the dietary (protein) adequacy decreases, however modeling challenges remain. Lastly, although convincing experimental data is not yet available, arguments based on modeling suggest that the risks of parasite evolution in response to genetic changes in the host should be less than the risks arising from other control strategies, such as anthelmintics. Thus, modeling techniques predict that selective breeding for resistance should be an effective and sustainable complementary control measure.     

Dose selection of selamectin for efficacy against adult fleas (Ctenocephalides felis felis) on dogs and cats

Selamectin, a novel avermectin, was evaluated in two controlled studies (one in Beagles, one in domestic shorthaired cats) to determine an appropriate topical dose for efficacy against adult Ctenocephalides felis felis (C. felis) fleas on dogs and cats for 1 month. For each study, animals were allocated randomly to four treatments. One treatment consisted of the inert formulation ingredients (vehicle) administered as a negative control, and the other three treatments consisted of a single topical dosage of 3, 6, or 9mgkg(-1) of selamectin. In each study, selamectin was administered as a topical dose applied to the skin in a single spot at the base of the neck in front of the scapulae. Dogs and cats were infested with 100 viable unfed C. felis (50 males and 50 females) on days 4, 11, 18, and 27. Seventy-two hours (+/-2h) after each infestation, on days 7, 14, 21, and 30, a comb count to determine the number of viable fleas present on each animal was performed. Efficacy of selamectin on day 30 was used to select an appropriate dose. For dogs and cats, percentage reductions in geometric mean flea comb counts for the three selamectin treatments ranged from 94. 6 to 100% on days 7, 14, and 21, compared with the negative-control treatment. On day 30, reductions in flea comb counts were 81.5, 94.7, and 90.8% for dogs, and 79.8, 98.0, and 96.2% for cats treated with selamectin at 3, 6, or 9mgkg(-1), respectively. For day 30 flea comb counts for dogs and cats, analysis of variance showed that the three selamectin treatments resulted in significantly (P< or =0.05) lower counts than did the negative-control treatment. For dogs and cats, geometric mean flea counts for selamectin administered at a dosage of 3mgkg(-1) were significantly (P< or =0.05) higher than those for the 6 and 9mgkg(-1) treatment dosages combined. There were no significant differences in flea counts between the 6 and 9mgkg(-1) treatments. This analysis was confirmed by linear-plateau modeling. Thus, the optimal dose of selamectin for efficacy against adult fleas for both dogs and cats, as estimated by the turning point (plateau) in the dose response curve, was 6mgkg(-1).     

Output of methylmercury from a Catchment in northern Sweden

The runoff output of methylmercury (MeHg) from the 50 ha Svartberget Catchment in northern Sweden was studied during 1993. These outputs are compared to those from the Gårdsjön Catchment in the southwestern part of Sweden. Although the wet deposition of MeHg is several times higher in southern Sweden the output of MeHg per unit area from the two catchments was comparable (0.12 g /km_* ²yr). Furthermore, the concentration of MeHg (0.4 ng/L) in the Svartberget Catchment was more than twice that from the Gårdsjön Catchment. These results suggest that the contemporary wet deposition of MeHg in itself is not a good indicator of runoff concentration or output per unit area. DOC transport and catchment characteristics such as wetland area, or possibly other forms of contemporary atmospheric deposition may all be more important for explaining MeHg output from the terrestrial ecosystem.     

Segregation of natural and experimental gastrointestinal nematode infection in F2 progeny of susceptible Suffolk and resistant Gulf Coast Native sheep and its usefulness in assessment of genetic variation

Gastrointestinal nematode parasitism is a concern to small ruminants worldwide. Productivity has been compromised because such nematodes, particularly Haemonchus contortus, have developed resistance to available anthelmintics. Some sheep breeds and lines within breeds are relatively resistant to infection, a trait that may be useful for developing control strategies. Suffolk sheep, which are susceptible to infection, were crossed with Gulf Coast Native sheep, which are more resistant to infection, to produce F1 progeny. F1 rams were bred to F1 ewes which produced 227 F2 offspring. These F2 offspring were evaluated for variability in infection levels, based on fecal egg count (FEC) and blood packed cell volume (PCV), under two natural infection conditions (one at weaning and another after a summer grazing period) and one experimental infection. The range of both FEC and PCV was large for all three infection periods with annual variation. Overall, the range for the three infection periods, respectively, were 167-149,933, 0-31,400 and 17-114,667 eggs per gram (EPG) of feces and 8.7-37.0%, 7.3-33.0% and 8.3-36.0%. This segregation of infection is what would be expected of F(2) progeny from susceptible and resistant parent breeds. Heritabilities of FEC and PCV for the three infection periods, respectively, were 0.15, 0.29 and 0.12, and 0.11, 0.22 and 0.12. Based on segregation of infection, larger heritabilities and maternal environment effects that declined after weaning, the summer natural infection was probably the best model for assessing genetic variation.     

Can scale-dependent landcover relationships explain canid community composition independent of intraguild occupancy?

Landscape Ecology - Niche theory is frequently used as a framework to integrate environmental variables and species interactions to describe species geographic distribution. Yet, the scale at which...     

Immunolocalization of Aquaporins 1 and 9 in the Ram Efferent Ducts and Epididymis

Aquaporins (AQPs) are essential membrane protein channels for the transport of water across membranes. Fluid movement in the epididymis is important for modulation of the luminal environment, in which sperm mature and reside. This study was designed to understand the morphology and localization of AQPs in ram efferent ducts (ED) and epididymis. For this purpose, the epididymis of seven animals were removed for histologic and immunohistochemical analyses. AQP1 immunoreactivity was observed in the apex of the ED, and AQP9 was found adjacent to the nuclei of the epithelial cells of the ED. The epithelial lining of ram epididymis is pseudostratified columnar and presents principal, basal, apical and narrow cells. In the initial segment (IS), a moderate reaction for AQP1 was observed in the apical cytoplasm of epithelial cells. An intense reactivity for AQP1 was noted over the microvilli of principal cells and in spermatozoa in the caput. In the corpus and cauda, AQP1 was noted only over the endothelial cells of vascular channels located in intertubular spaces. A weak‐to‐moderate reaction for AQP9 was observed in the nuclei of epithelial cells in the IS, caput and corpus of the epididymis. In the cauda, an intense reaction to AQP9 was observed in the epithelial border. In the IS, caput and corpus, the reactivity for AQP9 differed from those observed in domestic animals. The cauda showed a pattern similar to that previously described. These results indicate that AQPs 1 and 9 have reversed locations and roles in rams, suggesting activity variations related with fluid and solute absorption throughout the epididymis.