Cuticle formation in parasitic nematodes: RNA biosynthesis and control of molting.
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Specific antigen of G. spinigerum which has been shown to be a protein with a relative mol. wt of 24,000 (24K) was prepared from the advanced third-stage larvae (L3) obtained from the livers of naturally infected eels. The L3 were ground and extracted with water. Purification procedures involved gel filtration, chromatofocussing and anion exchange column chromatographies, while characterization of the specific antigen was performed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and staining, Western blot analysis and isoelectric focussing. The specific antigen which has a pI of 8.5 was used as antigen in the indirect enzyme-linked immunosorbent assay (ELISA) to detect specific antibody in four groups of individuals, namely five parasitologically diagnosed gnathostomiasis patients (group 1); 15 clinically diagnosed gnathostomiasis patients (group 2); 136 patients with other parasitic infections (group 3); and 25 normal healthy parasite-free controls. Sensitivity, specificity and predictive values (positive and negative) of the assay were 100%.
Adult Paragonimus heterotremus were recovered from the lungs and pleural cavity of cats orally infected with metacercariae. The worms were ground and extracted with distilled water. The soluble crude antigen (CA) contained about 40% proteins which could be fractionated by gel filtration on Sephadex G-200 into three profiles namely the F1, F2 and F3. The CA and its Sephadex profiles were used in an indirect enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to P. heterotremus in three groups of patients, i.e. patients whose sputum and/or faeces revealed P. heterotremus eggs (group 1), patients with other parasitic infections (group 2), bacterial proven tuberculosis patients (group 3) and healthy, parasite-free controls (group 4). The sensitivity and specificity of the assay when the F1 was used as the antigen were 100%. Western blot analysis revealed that specific antigen of P. heterotremus was a non-protein component of Mr35 kDa.
Age-related changes in the average worm burden and the prevalence of Trichuris trichiura infection, in a village community in St. Lucia, were examined by field studies based on worm expulsion techniques. Horizontal age-intensity profiles were convex in form with peak parasite loads occurring in the 2 to 15-year-old children. Prevalence is shown to be a poor indicator of changes in average worm load with age. Faecal egg counts (epg and epd) provide a qualitative measure of worm burdens since fecundity is shown to be approximately independent of worm load. The parasites were highly aggregated within the study community, with most people harbouring low burdens while a few individuals harboured very heavy burdens. Of the total parasite populations in the study sample, 84% were harboured by the 2 to 15-year-old children. Of those individuals harbouring 100 worms or more, 87% were in the 2 to 10-year-old age range. Crude estimates of population parameters (basic reproductive rate, 4-5; rate of reinfection, 90 year-1) suggest that the rate of reinfection is higher than for other helminth parasites of man. The control of morbidity and parasite transmission is discussed in the context of targeting drug treatment at the child segment of the study population.
In French Guiana, American cutaneous leishmaniasis is localized in the skin. The host response appears to be effective since few extra- or intracellular organisms can be found in tissue lesions, and we never observed any cutaneous dissemination or visceral involvement. However, this response is not fully effective since lesions may last for months. By using immunoperoxidase techniques and monoclonal antibodies directed against various cell populations, we examined the local immune response in skin biopsies. We found a high percentage of cells with the K/NK phenotype, a variable but usually high percentage of cells with the T cell phenotype bearing TAC receptors, and moderate numbers of monocytes and B cells. These results suggest that K/NK cells could play a role in the local control of parasite dissemination.
This paper discusses 2 prominent, contemporary issues in the epidemiological context of insect vector control: (i) the magnitude of the control problem, and (ii) non-linear processes influencing vector control. It concludes that we still cannot reliably measure the scale of some important control problems; e.g., there is considerable uncertainty about the basic reproduction number of malaria. The emergence of new concepts such as strain-specific immunity, and a growing emphasis on disease control as distinct from infection control, mean that some quantitative problems are being redefined more quickly than they are being solved. Population biologists have urged exploration of density-dependent processes which may help or hinder new methods of vector control. This brief review of non-linear phenomena such as facilitation and limitation finds little evidence that they will significantly influence, e.g., the introduction of some novel refractory mechanism into a vector population.
A high, constant and sustained release of drug appears to be a major requirement to avoid a late season rise in pasture infectivity that may result in production losses in calves in autumn or Type II osteragiasis in the following spring. The timing of administration is of crucial importance and will vary in regions of different epidemiology, such as between northern and southern United States. Lack of a standard and reliable technique for pasture larval counts has resulted in some negative or erratic results which are open to question. Although controlled release anthelmintics offer advantages of convenience, a comparison of the economic benefits at present favours prophylactic treatment of dairy heifers with conventional anthelmintics 3 and 6 weeks after spring turnout in northern regions of the Northern Hemisphere. There is a serious risk that boluses based on controlled release by diffusion will behave like slow decaying insecticides and select strongly for drug resistance, especially if farmers administer them in succession throughout the grazing season. There are, however, 2 features in the design of a controlled release device that in theory may minimize the risk of rapid selection for resistance: a high and constant release of anthelmintic followed by a rapid decline to zero as the device becomes exhausted. Under these conditions, the device may even prolong the useful life of an anthelmintic to which resistance has already developed. This paper was presented at Pfizer Symposium on The Application of Sustained Release Anthelmintic Dosage Forms in the Control of Parasites in Grazing Animals at the World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) 10th International Conference, 18-20 August 1983, Perth, W.A., Australia.
Herbage intake was measured in two groups of 20 first-year grazing cattle. The animals in one group each received a morantel sustained release bolus at turnout to control nematode parasitism and the animals in the other group remained untreated. The latter group showed a mean peak faecal egg count of 655 eggs per gram (e.p.g.) in October associated with high serum pepsinogen concentration and clinical signs of ostertagiasis, compared with a peak of 119 e.p.g. in the treated group which remained in good health. In September the daily voluntary feed intake of the untreated animals was significantly depressed (94 g kg-1 body weight vs. 77 g kg-1 P less than 0.001), but no difference in digestive efficiency was recorded between the two groups. This difference in feed intake was associated with a 47 kg mean live weight advantage of the treated animals at housing.
Lambs were challenged by dosing with 2500 T. colubriformis larvae per day for 34 weeks, rested for 4 weeks and then re-challenged with the infective larvae for a further 10 weeks. A technique for the measurement of inhibition of larval migration from agar gels was used to investigate the antiparasitic activity of mucus, obtained from the small intestine and abomasum of the lambs, at the end of the re-challenge period. Measurements were also made on ileal digesta samples collected at different times during the development of the initial resistance, the rest period and the re-challenge period, and on faeces collected during the re-challenge infection. The mucus from the small intestine and abomasum paralysed and inhibited larval migration from agar gels significantly more (P less than 0.01 and P less than 0.05, respectively) than corresponding mucus from parasite-free control animals. This inhibitory activity was also detected (P less than 0.01) in the ileal digesta of infected animals from Week 6 of primary dosing. The magnitude of the inhibition in the ileal digesta increased with time during the infection period and was again detected during re-infection (P less than 0.01). It was not detectable in resistant sheep towards the end of the rest period. Slight inhibitory activity was detected in faeces after 2 weeks of re-infection. These observations suggest that substances secreted into the lumen of the small intestine of infected animals are responsible for resistance against ingested larvae.
Avermectins, a relatively new class of broad spectrum pesticides, are used widely to control livestock parasites. Following treatment, avermectins are eliminated in the livestock faeces where they also have a wide range of harmful affects upon certain characteristic insects that breed in dung, few of which are pests, and many of which are beneficial. The effects range from acute toxicity in larvae and adults, through disruption of metamorphosis, to interference with reproduction. Different methods of drug administration lead to different concentrations of drug residues in the faeces, which in turn influence the responses of non-target organisms. Higher Diptera are particularly sensitive to drug residues and show a wide range of responses from death of larvae to developmental abnormalities in the adults. Larvae and immature adults of Coleoptera show some mortality in the dung of recently treated animals, while delayed effects upon reproduction and physiology have been observed in adults feeding on dung at longer post-treatment times. Although the impact of lethal doses has been described in some species, the effects of sub-lethal doses have hardly been recognised at the present time. Correlated with the deleterious effect upon dung-breeding insects, a retardation in the rate of loss of biomass of dung pats from avermectin-treated cattle has been observed following the various forms of drug administration. Differences in methodology, inappropriate statistics, and/or extremes of climatic conditions prevailing at the time of testing, explain the results of those studies where such delays have not been observed. It is short-sighted to consider only dung dispersal in relation to avermectin usage, a practice that overlooks the impact on the insects themselves and their diverse roles in pastureland ecology.
Scarabaeine dung beetles feeding on dung from cattle treated with an injection of avermectin at a therapeutic dose to control internal parasites, show larval mortality, mortality of immature adults, reduced egg production, and inhibited ovariole development for periods of 1-4 weeks following treatment. In winter rainfall regions of Australia, feeding by newly emerged adults of an introduced species, Onthophagus binodis, resulted in shredding of cattle dung between December and May. Production of brood masses and eggs resulted in dung being buried from September to November. Feeding by newly emerged adults of a native species, Onthophagus ferox, resulted in dung being buried in May and June, and production of brood masses for breeding resulted in dung burial between September and November. There are thus 2-6 months of the year when injection of cattle with avermectin would affect mortality of newly emerged beetles, and 3 months of the year when avermectin treatment of cattle would affect dung beetle oviposition or larval survival. Beetles were attracted to fresh dung for 2-4 days, and most had left the pad within a week. There are no data to indicate the effects of avermectin residues in dung on dung beetle populations and on beetle fitness. Tests should be done to determine if avermectins in slow release devices have a greater effect on dung beetles than injections, and to determine what effects both have on dung beetle populations.
We have isolated a cDNA from the protozoan parasite Leishmania major (Lm) that encodes a protein homologous to the Saccharomyces cerevisiae and Kluyveromyces marxianus silent information regulator 2 (SIR2) proteins. The deduced Lm SIR2-related protein (termed LmSIR2rp) consists of 381 amino acids that share 40.5% identity with yeast SIR2, increasing to 60% when substitutions are included. Moreover, the LmSIR2rp aa sequence contains a single potential zinc-binding domain with a CysXaa2CysXaa20CysXaa2Cys motif, and its C-terminal part is rich in Ser (16 Ser residues over 40 aa) which constitute potential sites for phosphorylation. The characterization of a novel Lm gene product which shows considerable similarity to a yeast mating-type regulatory protein provides a new tool to investigate the parasite differentiation control mechanisms and gene expression regulation.
Caenorhabditis elegans represents an excellent model in which to dissect the biosynthesis and assembly of the nematode cuticle. A sequenced genome, straightforward transgenesis, available mutants and practical genome-wide RNAi approaches provide an invaluable toolkit in the characterization of cuticle components. We have performed a targeted RNAi screen in an attempt to identify components of the cuticle collagen biosynthetic pathway. Collagen biosynthesis and cuticle assembly are multi-step processes that involve numerous key enzymes involved in post-translational modification, trimer folding, procollagen processing and subsequent cross-linking stages. For many of these steps, the modifications and the enzymes are unique to nematodes and may represent attractive targets for the control of parasitic nematodes. A novel serine protease inhibitor was uncovered during our targeted screen, which is involved in collagen maturation, proper cuticle assembly and the moulting process. We have confirmed a link between this inhibitor and the previously uncharacterised bli-5 locus in C. elegans. The mutant phenotype, spatial expression pattern and the over-expression phenotype of the BLI-5 protease inhibitor and their relevance to collagen biosynthesis are discussed.
OBJECTIVE: To review evaluated studies of health education within health facilities in developing countries. METHOD: Extensive searchers were carried out of electronic databases and other sources to identify published evaluations of heath education within health service settings in developing countries. Those reports providing evidence of impact were selected for further consideration and included in the review. RESULTS: Examples of successful practice are presented for a wide range of health topics that include family health, antenatal care, nutrition education, reduction of use of injections, improve adherence to regimes for antibiotics and other, prevention and control of parasitic and infectious diseases, reproductive health including AIDS and sexually transmitted diseases. CONCLUSIONS: There are now many examples of effective patient education within the challenging situations found in the developing world. PRACTICE IMPLICATIONS: Strategies adopted by successful patient education programmes are listed including initial needs research, training of staff in communication skills, cultural sensitivity, including family members, using lay volunteers, development of improved packaging for medicines, clear written instructions with simple (and pre-tested) pictorial advice, participatory learning methods, and patient self-help groups, mail reminders and reminder stickers, practical demonstrations and more effective use of waiting areas including use of video.
At present, there is no effective alternative to chemical control of parasitic helminths where livestock are grazed intensively. Resistance to anthelmintics has become a major problem in veterinary medicine, and threatens both agricultural income and animal welfare. The molecular and biochemical basis of this resistance is not well understood. The lack of reliable biological and molecular tests means that we are not able to follow the emergence and spread of resistance alleles and clinical resistance as well as we need. This review summarizes some of the recent findings on resistance mechanisms, puts forward some recommendations for limiting its impact and suggests some priorities for research in this area.
More than 20 years after Dawkins introduced the concept of "extended phenotype" (i.e. phenotypes of hosts and parasites result from interactions between the two genomes) and although this idea has now reached contemporary textbooks of evolutionary biology, most studies of the evolution of host-parasite systems still focus solely on either the host or the parasite, neglecting the role of the other partner. It is important to consider that host and parasite genotypes share control of the epidemiological parameters of their relationship. Moreover, not only the traits of the infection but also the genetic correlations among these and other traits that determine fitness might be controlled by interactions between host and parasite genotypes.
In Cambodia, schistosomiasis is transmitted in the provinces of Kratie and Stung Treng where approximately 80000 individuals are estimated to be at risk of infection. The baseline prevalence of infection was estimated to be between 73% and 88%, and cases of severe morbidity (hepatosplenomegaly, puberty retardation) and mortality were very common. In 1994, the Ministry of Health of Cambodia started schistosomiasis control applying universal chemotherapy with praziquantel (40mg/kg). The coverage of the programme was between 62% and 86% for 8 years. This simple control measure resulted in the control of the disease: no cases were reported in 2004 and only three cases were reported in 2005. In addition, there are no longer reports of cases of severe morbidity due to schistosomiasis. Since the beginning of the control programme, a single dose of mebendazole (500mg) has been combined with praziquantel during the mass chemotherapy; as a result the prevalence of Ascaris lumbricoides and hookworms dropped from 74.5% to 10% and from 86% to 40% respectively. The experience in Cambodia demonstrates that, with political commitment, control of parasitic diseases is achievable even in a situation of minimal resources. The programme represents a successful model for other developing countries.
Lernaea cyprinacea infestation is a major problem for fishing culture in Brazil, which was introduced in the mid-eighties. To attempt controlling this parasite, an evaluation of the activity of the Pinus elliottii resin constituents against it was undertaken. To run the bioassay, fish infested with L. cyprinaceae were transferred to aquaria and kept at room temperature and ventilation for 15 days for adaptation. Afterwards, fish were sacrificed and the parasites were kept under water to run the experiments by evaluating the activities of the steamed oil and the chloroform fraction of the resin at concentrations of 0.5, 1.0, 2.0 and 5.0 ppm. The crude resin was also evaluated at concentrations of 1.0, 5.0 and 10 ppm, as well as the major components of its steamed oil, alpha and beta-pinenes, both at concentrations of 0.5 and 1.0 ppm. The results showed that both fractions from the resin were effective at 0.5 ppm concentration, while the pure compounds were less active. Moreover, the acute toxicity (DL(50)) of the crude resin for the fish Leptorinus piau, aged 1 month, was established at 200.0 ppm, which ensures its safe use.