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Induction of traps by Ostertagia ostertagi larvae, chlamydospore production and growth rate in the nematode-trapping fungus Duddingtonia flagrans.

Biological control of parasitic nematodes of domestic animals can be achieved by feeding host animals chlamydospores of the nematode-trapping fungus Duddingtonia flagrans. In the host faeces, D. flagrans develop traps that may catch nematode larvae. In experiments on agar, D. flagrans had a growth rate between 15 and 60 mm/week at temperatures between 20 and 30 degrees C. The presence of nematodes induces the fungus to produce traps. The rate of trap formation in D. flagrans has an optimum at 30 degrees C, producing 700-800 traps/cm2/2 days, when induced by 20 nematodes/cm2 on agar. Approaching 10 and 35 degrees C the ability to produce traps is gradually reduced. The response of chlamydospore production on agar to changes in temperature is the same as that for trap formation. On agar, at 10, 20 and 30 degrees C D. flagrans loses its trap inducibility after 2-3 weeks. During the ageing process, increasing numbers of chlamydospores are produced up to a certain limit. The time for reaching maximum chlamydospore concentration coincided with the time for loss of induction potential. The implications of these results in relation to biological control in faeces are discussed.

Animals↗

The impact of internal parasites on the productivity of young cattle organically reared on semi-natural pastures in Sweden.

A grazing experiment with young cattle was conducted over two consecutive (1997, 1998) grazing seasons on semi-natural pasturelands in central-eastern Sweden. Comparisons were made between groups of animals that were either untreated and set-stocked, ivermectin bolus treated and set-stocked or untreated but moved in mid-summer (mid-July) to ungrazed pasture. The whole experimental area had remained virtually free of cattle during the previous two seasons and the cattle had been raised indoors since birth. To introduce low-levels of parasite infection into the experimental system, each animal received a 'priming dose' of approximately 10, 000 infective trichostrongylid larvae at the time of turnout for both years. Results of the first year study showed that the level of parasitism was so low that it failed to induce any productivity losses in both groups of untreated cattle, which grew as well as those given boluses at turnout. In contrast, in 1998 both groups of untreated cattle suffered varying degrees of sub-clinical and clinical parasitism to result in an average of 30kg liveweight depression, compared with the bolus treated cattle, at the end of the season. The only major departure between the two years was that in the latter, the cattle in the untreated groups were exposed to infective larval pickup, which had overwintered on pasture. Cattle in the move treatment grazed in the same sequence on pastures used by similar classes of animals during the previous year. That is, their pastures at turnout had not been grazed since mid-summer of the previous year. Clearly this early season (1997) grazing by young cattle resulted in sufficient overwintered larvae at the start of the following year (1998) to cause productivity losses of the same magnitude as those recorded for young cattle grazing on pastures contaminated for the entire grazing season of the previous year. This was confirmed by tracer tests that were carried out on all treatments, at the time of turnout and the mid-summer move in 1999. These results have major significance to organic cattle producers in Sweden who have a much higher tendency to practice a variety of grazing management techniques aimed at controlling nematode parasite infections in young cattle, than their conventional farming colleagues. It has been identified that one of these strategies is to simply use summer/autumn saved pastures for young stock at turnout, which if grazed by young stock prior to this, could prove to be counter-productive.

Animal Husbandry↗

USAID in Nepal.

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Agriculture↗

Effects of praziquantel on experimental Schistosoma bovis infection in goats.

The effect of praziquantel against experimental Schistosoma bovis infection in West African Dwarf goats was investigated. Thirty goats were exposed to 2000 cercariae each and 15 of those received a praziquantel treatment (60 mg kg-1) 13 weeks post-infection. One day, 1 week and 4 weeks post-treatment representative goats from each group were killed and worms were recovered by perfusion. For comparison, parasite-free control animals were monitored, some of which were given praziquantel. Every second week during the study, faecal samples were collected. The cure rate was 100% 1 day, 99.4% 1 week and 95.7% 4 weeks post-treatment. Tissue egg counts were significantly reduced (P < 0.001) 4 weeks post-treatment in all parts of the intestines, but not in the liver. Faecal egg counts were reduced by 84.1% 1 week and by 98.3% 3 weeks after treatment, the reduction being highly significant both 1 week 3 weeks after treatment (P < 0.001). Overall strong correlations between the number of worm pairs, tissue egg counts and the final faecal egg count were observed, indicating that the faecal egg counts during infection and following treatment can be used as a guideline for the pathology associated with the infection.

Animals↗

Pharmacokinetics of doramectin and ivermectin after oral administration in horses.

A study was undertaken in order to compare plasma disposition kinetic parameters of doramectin (DRM) and ivermectin (IVM) in horses after oral administration. Ten crossbreed adult horses, clinically healthy, weighing 380-470 kg body weight (bw) were selected for study. Faecal examinations were performed to determine faecal parasite egg counts. Horses were allocated to two groups of five animals to provide an even distribution considering the variables sex, body weight and faecal egg count. Group I, were treated with an oral paste formulation of IVM at 0.2 mg/kg b/w and Group II, were treated with an oral dose of 0.2 mg/kg bw of DRM prepared as paste from the injectable formulation for oral administration. Blood samples were collected by jugular puncture between 0 h and 75 days post-treatment. Plasma was separated and later solid phase extraction and derivatization samples were analysed by high performance liquid chromatography (HPLC); a computerised kinetic analysis was carried out. Data were compared using the Mann-Whitney U-test. The mean plasma concentrations of DRM and IVM after oral administration in horses were detected until 30 and 20 days, respectively. Both drugs showed similar patterns of absorption and no significant differences were found for peak concentration, the time to peak concentration, or for absorptive half-life. The terminal elimination half-life was significantly (P<0.05) longer in the DRM treated group than for the IVM treated group. The differences observed in the elimination half-life explain the longer mean residence time and high values of area under the concentration time curve for the group treated with DRM, which are 30% higher than those of the IVM group. Considering its pharmacokinetics, tolerance and anthelmintic efficacy, the oral administration of DRM, could be an alternative to IVM for the control of parasitic diseases of horses.

Administration, Oral↗

Linker scanning mutagenesis of the Plasmodium gallinaceum sexual stage specific gene pgs28 reveals a novel downstream cis-control element.

Protozoan parasites undergo complex life cycles that depend on regulated gene expression. However, limited studies on gene regulation in these parasites have repeatedly shown characteristics different from other eukaryotes. Within the Apicomplexa family, little is known about the mechanism of gene expression and regulation in Plasmodium spp. We have been investigating the cis-elements that control basal expression of a sexual stage specific gene in Plasmodium gallinaceum. Previously, we identified by 5' deletion analysis of a reporter construct that the 333bp upstream of the translational start site of pgs28 is sufficient for basal expression, and that the sequence between -333 and 316bp is necessary for such expression. In this report, we identified by linker scanning mutagenesis an 8-bp sequence that is essential for pgs28 transgene expression. This sequence is a target of sequence-specific nuclear factors. Primer extension studies demonstrate that, interestingly, the endogenous pgs28 transcript has two 5' ends, at -65 and +1. We suggest that this 8-bp sequence, CAGACAGC that is situated at +24 to +31 (with respect to the proximal start site), is a novel downstream promoter element in P. gallinaceum that appears to function independently of a TATA box or an Inr element.

Animals↗

Cryptosporidium parvum-specific mucosal immune response in C57BL/6 neonatal and gamma interferon-deficient mice: role of tumor necrosis factor alpha in protection.

Both neonatal and C57BL/6 gamma interferon (IFN-gamma) knockout (C57BL/6-GKO) mice are susceptible to Cryptosporidium parvum, but the course of infection is different. Neonatal mice are able to clear the parasite within 3 weeks, whereas C57BL/6-GKO mice, depending on age, die rapidly or remain chronically infected. The mechanism by which IFN-gamma leads to a protective immunity is yet poorly understood. In order to investigate the effect of IFN-gamma on other cytokines expressed in the intestinal mucosa during C. parvum infection, we studied cytokine mRNA expression in the neonates and GKO (neonatal and adult) mice by quantitative reverse transcription-PCR (RT-PCR) at 4 and 9 days after infection. IFN-gamma mRNA levels were quickly and strongly up-regulated in the mucosa of neonatal mice. In GKO mice, the Th1-type response was dramatically altered during the infection, whereas the mRNA expression levels of the Th2-type cytokines interleukin 4 (IL-4) and IL-10 were increased in both mouse models. In the absence of IFN-gamma, the adult knockout mice up-regulated the mRNA levels of inflammatory cytokines, such as IL-1beta, IL-6, and granulocyte-macrophage colony-stimulating factor, in the mucosa, but not tumor necrosis factor alpha (TNF-alpha), whereas all these cytokines were up-regulated in the infected neonatal mice. Further experiments indicated that injections of TNF-alpha into GKO adult mice significantly reduced oocyst shedding. The results of the present study indicate that the resolution of infection is dependent on the expression of Th1-type cytokines in the mucosa of C57BL/6 mice and that TNF-alpha may participate in the control of parasite development.

Animals↗

Affinity purified oval antigen for diagnosis of Opisthorchiasis viverrini.

Monoclonal antibodies (MAb) were raised against an oval antigen of the liver fluke Opisthorchis viverrini which is the causative agent of a parasitosis, i.e. opisthorchiasis in Thailand. The antibodies were used in an affinity column to purify the O. viverrini oval antigen from a crude extract of adult parasites by chromatography. The oval antigen was then used in a membrane (dot) ELISA for detecting antibodies in serum samples of parasitologically confirmed Opisthorchis viverrini infected individuals (adult parasites were found in stools after praziquantel treatment and salt purgation), as well as of individuals infected with other parasites and parasite-free controls. The MAb-based dot-ELISA using the affinity purified O. viverrini oval antigen revealed 100% sensitivity, specificity and accuracy for detecting O. viverrini infection. The test is simple, rapid and highly reproducible. Several samples can be tested at the same time without the requirement for special equipment or much increase in testing time; thus it is suitable for mass screening for O. viverrini exposure, especially in new endemic areas. Furthermore using serum specimens could increase patient and community compliance compared to the conventional parasitological survey which uses stool samples for the detection of O. viverrini ova, without treatment and subsequent salt purgation, this conventional method shows a low sensitivity and is also unpleasant to both the sample donors and the laboratory technicians which has historically shown a further negative impact on the final outcome.

Animals↗

Effect of level of feed intake on the development of gastrointestinal parasitism in growing lambs.

A study was conducted to examine the role of nutrition in the development of gastrointestinal (GI) parasitism, performance and pathophysiology of parasitism in female lambs. Forty-four months old ewe lambs received for 6 weeks 0, 1500 or 7000 L(3) larvae of Teladorsagia circumcincta in two doses per week. The animals were given access to a pelleted diet to meet energy requirements for gaining weight close to their potential (H) or approximately 50% of it (L). The level of protein in the diet was 20-30% above requirements for both planes of nutrition. Measurements of body weight, serum pepsinogen concentration and faecal egg counts were taken at weekly intervals. All animals were slaughtered 2 weeks after the last larval dosing for estimation of worm count, carcass composition and reproductive tract development. Voluntary intake of infected animals decreased by approximately 10%. Both nutrition and infection influenced lamb performance and carcass composition. However, no direct effects on reproductive tract parameters due to the presence of infection were observed. The level of L(3) infection showed a significant effect not only on faecal egg counts and on worm burden, as it could be expected, but also on the proportion of females to males, which increased significantly with the level of infection. While the plane of nutrition did not show a significant effect on faecal egg counts or on worm burden, surprisingly, both female worm size and their fecundity decreased significantly with the level of nutrition. This response was accompanied by a significant increase in the concentration of circulating eosinophils suggesting that the immune response of lambs consuming high levels of energy was enhanced compared to that of lambs kept on a restricted diet. Results herein show that in young female lambs fed on adequate levels of protein, an improvement in energy supply does not only improve carcass characteristics but clearly enhances the development of resistance to GI nematodes infection. This may have decisive management implications for the control of parasitic infections in sustainable production systems.

Animals↗