Diversity,seasonality and sites of attachment of adult ixodid ticks on dogs in the central region of the Free State Province,South Africa

Several surveys of ticks infesting dogs belonging to owners in resource-limited and more affluent communities have been conducted in South Africa, but no such investigation has been carried out in the Free State Province of this country. The present study was initiated to meet this shortcoming. Ticks were collected from dogs at six localities in, and to the east of the city of Bloemfontein in the central region of the province. Three of these localities could be classed as resource-limited and two as affluent, while the sixth locality was an animal shelter serving all members of the public. Adult ticks belonging to nine ixodid tick species were collected, of which Rhipicephalus sanguineus was the most numerous. Significantly more R. sanguineus was collected from dogs at resource-limited than at more affluent localities. The greatest proportions of these ticks attached to the backs and necks of the dogs, with the proportions being larger in long-haired than in short-haired dogs. Most R. sanguineus were collected during the warmer months particularly from January to April. The greatest proportions of Haemaphysalis leachi, the next most numerous species, were also collected from the backs and the necks of the dogs. Most of these were present during the period September to November.     

Determination of transpiration in irrigated grapevines: comparison of the heat-pulse technique with gravimetric and micrometeorological methods

The use of a heat-pulse technique to monitor sap flow from which transpiration can be deduced was evaluated in ungrafted grapevines (Vitis vinifera L., cv. Sultana) under glasshouse and field conditions. There was a significant degree of agreement between daily transpiration deduced from heat-pulse velocity (T _hp) and that determined directly by gravimetry in the glasshouse and by calibration using the Penman-Monteith equation in the field. Comparison throughout the growing season of T _hp to transpiration calculated with the full Penman-Monteith equation produced a high coefficient of determination (r ²=0.69). A similar comparison of T _hp with transpiration calculated with only the aerodynamic component of the Penman-Monteith equation produced a non-linear relationship, due to the equation over-estimating transpiration relative to T _hp at high vapour pressure deficits (i.e. above 2.5 kPa). Values for total seasonal transpiration measured with heat-pulse sensors or calculated with either the full Penman-Monteith equation or with only the modified aerodynamic component of the equation were within 10% of each other. Transpiration (T _hp) in grapevines with an adequate supply of soil water was shown to be coupled to ambient air conditions. Received: 20 July 1998     

Earthworm ecological groupings based on C analysis

We report the first use of ¹⁴C isotope analysis to investigate the ecological grouping of earthworms. Mature endogeic (Allolobophora caliginosa), mature epigeic (Lumbricus rubellus), and semimature anecic worms (A. longa) were collected in September 2002 from a woodland site at Lancaster, UK. Because anecic worms are known to have a variable feeding behaviour and can show dietary changes during ontogeny, additional immature and mature specimens of A. longa were also collected from the same site in January 2004. Epigeic earthworms showed the lowest radiocarbon concentration (0-3-years old), implying that they assimilated more recently fixed carbon than the anecic or endogeic earthworms. The age of carbon assimilated in mature anecic species (5-7-years old) was closer to that of endogeic species (5-8-years old) than to epigeics, suggesting that a greater proportion of older, more mineralised organic matter may form part of the diet of the anecic earthworms than previously thought. These results suggest that ¹⁴C approaches are useful in the study of the feeding behaviour of detritivorous animals by providing in situ information on the age of the carbon assimilated by the worms. This can then be related to their role in ecosystem functioning, particularly in carbon cycling.     

Cytokines in mammary lymph and milk during endotoxin-induced bovine mastitis

Cytokine kinetics were examined in milk and in afferent and efferent lymph of the supramammary lymph node after intramammary infusion of endotoxin from Escherichia coli. Cows were sampled 0, 2 and 4h after infusion (p.i.). Neutrophils appeared in afferent lymph 2h p.i., and in efferent lymph and milk 4h p.i. The milk contained high concentrations of interleukin-8 (IL-8) at 2 and 4h p.i. IL-8 was also found in lymph, but at lower concentrations. The tumor necrosis factor-alpha (TNF-alpha) concentration tended to increase in afferent lymph at 2h p.i., and increased in milk at 4h p.i. The level of IL-1beta increased at 4h p.i. in milk, but was not detected in lymph. Interferon-gamma was not detected in any sample, at any time. The results indicate a primary role for IL-8 in the recruitment of neutrophils into the gland, and suggest that IL-1beta and TNF-alpha are not necessary for IL-8 production and release in response to endotoxin.     

Enzymeimmunoassay of oestrone sulphate concentrations in faeces for non-invasive pregnancy determination in mares

AIM: To determine the suitability of measuring faecal oestrone sulphate (OS) by enzymeimmunoassay as a means of determining pregnancy status in mares bred under New Zealand conditions. METHODS: An antibody-coated microtitre plate-based enzymeimmunoassay was used to determine the concentration of OS in faecal and plasma samples obtained from pregnant and non-pregnant mares. RESULTS: In non-pregnant mares, the mean faecal OS concentration was 34 ng/g, and the value three standard deviations above this was 80 ng/g. None of 427 faecal samples collected from 116 non-pregnant mares over a l-year period had an OS concentration >80 ng/g. Only five samples from three mares had an OS concentration >65 ng/g, the value two standard deviations above the mean non-pregnant value. Analysis of faecal OS concentrations in 532 faecal samples collected from 39 pregnant mares showed that as pregnancy progressed, an increasing proportion of faecal samples had OS concentrations >80 ng/g. None of the mares 150 days or more pregnant had faecal OS concentrations <50 ng/g, and 204/220 samples obtained from these mares had faecal OS concentrations >80 ng/g. Following foaling or foetal death, elevated faecal OS concentrations returned quickly to non-pregnant levels. The mean +/- s.e.m. plasma level of OS in five mares bled daily throughout one oestrous cycle was 1.7 +/- 0.2 ng/ml. Sixty-eight blood samples from pregnant mares bled up to five times between 92 days after mating and foaling all had plasma OS concentrations >30 ng/ml, with 64/68 being >50 ng/ml. CONCLUSIONS: This study shows that measuring faecal OS concentrations by enzymeimmunoassay offers a convenient, accurate, non-invasive means of determining pregnancy status in mares from 150 days after mating onwards. Mares with faecal OS concentrations <50 ng/g can be considered not pregnant, while mares with faecal OS concentrations >80 ng/g can be considered pregnant. Those few mares returning a faecal OS concentration between 50 and 80 ng/g should be retested to obtain a conclusive result. Measuring plasma OS concentrations allows pregnancy status to be determined earlier (from 100 days after mating). Moreover, the discrimination between non-pregnant and pregnant levels is greater for OS in plasma than in faeces. CLINICAL RELEVANCE: Measurement of OS concentrations in faeces provides an alternative and non-invasive means of determining pregnancy status in mares from 150 days after mating.     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Cytokine response of porcine cell lines to Salmonella enterica serovar typhimurium and its hilA and ssrA mutants

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular bacterium which can infect and colonize pigs. After contact with enterocytes and macrophages, S. Typhimurium induces production of cytokines thus triggering the innate immune response. In this study we evaluated the cytokine response of two porcine cell lines, IPI-2I and 3D4/31, of epithelial or macrophage origins, respectively, to the wild-type S. Typhimurium and its hilA and ssrA mutants. We observed that the 3D4/31 cell line essentially did not respond to S. Typhimurium infection when a medium with foetal calf serum was used. However when the 3D4 cell line was incubated overnight in the presence of porcine serum, it efficiently responded to the wild-type strain and the ssrA mutant but not to the noninvasive hilA mutant as measured by mRNA quantification of TNF-alpha, IL-8 and GM-CSF by the real-time RT-PCR. In IPI-2I, all the cytokines were also induced by the wild-type S. Typhimurium and the ssrA mutant although the induction of TNF-alpha was lower than that induced by the wild-type strain. The hilA mutant was unable to induce any of the cytokines tested. The ssrA mutant can therefore be considered as more suitable for further vaccine development as the stimulation of innate immune response is important for animal protection against a challenge with virulent strains.     

Enzyme-linked immunosorbent assay for antibodies to Campylobacter fetus in bovine vaginal mucus

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to Campylobacter fetus subspecies venerealis in bovine vaginal mucus. The results of testing 168 samples from experimentally infected, field cases and control cows showed that the ELISA was more sensitive than the vaginal mucus agglutination test and also detected antibodies in earlier stages of infection.