This discussion covers the safety and clinical toxicity of three anthelmintics of the tetrahydropyrimidine class of neuroactive agents, compounds that are primarily active against a range of nematodes affecting humans and animals. These agents act by selectively targeting the nervous system of the worms by inhibiting the neurotransmitter acetylcholine system, and thus interfering with the worms' ability to remain in the host. Side effects and intoxication, when they result, are most commonly due to the direct effect of these agents on this nervous system in the host. Nevertheless, in general, these agents remain amongst the safest of anthelminthic drugs in use today. The clinical changes related to toxicity with all three drugs include effects on the intestinal physiology and function (nausea, abdominal discomfort, and diarrhea), CNS changes (dizziness and nausea), and less commonly, changes in other body systems. These agents are commonly used in both veterinary and human medicine successfully and safely, and often in combination with other anthelmintics.

The identification of excreted–secreted (ES) proteins of filarial nematodes as potential diagnostic reagents is an important requirement for the development of methods to determine level of infection in the host, especially for human filariae. Dirofilaria immitis, the canine heartworm, is a widespread and important veterinary pathogen and is a useful model for filarial parasites of humans. An analysis of proteins released from adult D. immitis (the secretome) in culture is available. We sought to identify D. immitis ES proteins found in vivo to validate the in vitro secretome and to investigate them as potential diagnostic reagents. Cultures of D. immitis adults obtained from infected dogs were maintained for 72 hr with daily changes of media. Proteins were concentrated from spent media by standard methods and were passed through Protein-A columns containing purified IgG antibodies from heartworm-infected dogs. Following extensive washing, heartworm proteins recognized by the antibodies were eluted from these columns and submitted for analysis by tandem mass spectrometry (MS/MS). As a comparison, somatic proteins from adult D. immitis female parasites and microfilaria were also processed and analyzed by the same protocol. Six, 9, and 12 proteins were identified by MS/MS in the ES, adult female, and microfilaria samples, respectively. The identification of the most abundant parasite proteins present in the serum of infected hosts offers a rational approach to the development of new diagnostic assays that may be applicable across the Filarioidea.

The identification of excreted-secreted (ES) proteins of filarial nematodes as potential diagnostic reagents is an important requirement for the development of methods to determine level of infection in the host, especially for human filariae. Dirofilaria immitis, the canine heartworm, is a widespread and important veterinary pathogen and is a useful model for filarial parasites of humans. An analysis of proteins released from adult D. immitis (the secretome) in culture is available. We sought to identify D. immitis ES proteins found in vivo to validate the in vitro secretome and to investigate them as potential diagnostic reagents. Cultures of D. immitis adults obtained from infected dogs were maintained for 72 hr with daily changes of media. Proteins were concentrated from spent media by standard methods and were passed through Protein-A columns containing purified IgG antibodies from heartworm-infected dogs. Following extensive washing, heartworm proteins recognized by the antibodies were eluted from these columns and submitted for analysis by tandem mass spectrometry (MS/MS). As a comparison, somatic proteins from adult D. immitis female parasites and microfilaria were also processed and analyzed by the same protocol. Six, 9, and 12 proteins were identified by MS/MS in the ES, adult female, and microfilaria samples, respectively. The identification of the most abundant parasite proteins present in the serum of infected hosts offers a rational approach to the development of new diagnostic assays that may be applicable across the Filarioidea.

Immunocytochemical examination of onchocercal nodule tissues containing adult Onchocerca volvulus using immuno-markers for blood and lymphatic vessels (vWF, D2-40, podoplanin, Prox-1, and Lyve1) shows a distinct pattern of distribution of these vessels within nodules. Blood vessels were commonly seen associated with organized lymphoid cellular aggregates in the both the outer and inner areas of the nodules. In contrast, the majority of the lymphatic vessel positivity was seen in the central zone in close apposition to the adult parasites, and the remainder usually associated with microfilariae in the outer areas of the nodule. These findings suggest an intimate relationship between adult O. volvulus and lymphatic vessels, including the likely proliferation of lymphatic endothelial cells (lymphangectasia) akin to that seen with other filariae. These findings indicate that adult O. volvulus may migrate via the lymphatic system, and that clinical manifestations of this disease that involve tissue edema may be the result of the location of these worms in the lymphatic system.

The Tanzania Lymphatic Filariasis Programme, which was launched in 2000, is, in terms of geographical coverage, among the largest disease-control programmes in Tanzania's history, currently reaching 9.4 million people in 34 districts. The issues associated with this programme's implementation are reviewed here, in the context of the various players/stakeholders involved. This article provides an insight of how the programme began and discusses key areas in the programme's design. Mainly, however, it gives some impressions of how the programme is perceived by, and how it affects, village healthworkers, patients and politicians - the people who contribute to the implementation of the programme at various levels.

The defining images of lymphatic filariasis are the horrendous disfigurements of lymphoedema, elephantiasis and hydrocele. These clinical presentations, although obviously important and life changing, are not, however, the only outcomes of this wide-spread filarial infection. The other effects of the disease range from severe, acute but short-term bouts of sickness to psychological impairment, poverty and family hardship. It is important to support cases of the disease through all means available, such as reparative hydrocelectomy, hygiene training and facilitation, and the provision of adequate chemotherapy. Although only a minority of the residents in any endemic community is affected with the severe clinical manifestations of this parasitic infection, these cases are central to, and important advocates for, the current global effort to eliminate the infection through mass drug administrations (MDA). Their clinical improvement acts as an important catalyst for the general population and encourages high compliance in the MDA.This communication discusses the central role that filariasis patients have played in the Tanzania Lymphatic Filariasis Elimination Programme to date, and covers some of the clinical successes achieved in the past 10 years. The abolition of the clinical manifestations of filarial infection remains the ultimate goal of the Global Programme to Eliminate Lymphatic Filariasis, and maintaining a focus on the affected individuals and their clinical condition is vital to that programme's overall success.

Dental waterlines contain large numbers of Gram-negative bacteria. Endotoxin, a component of such organisms, has significant health implications. Paired samples of dental unit water and the aerosols generated during dental procedures were collected, and assayed for bacteria and endotoxin levels, using heterotrophic plate counts and the Limulus amoebocyte lysate test. Consistent with published studies, the extent of bacterial contamination in the dental waters sampled for this investigation surpassed the levels associated with potable water, with counts in excess of 2.0x10(6) c.f.u. ml(-1) in some samples. Correspondingly high concentrations of endotoxin [up to 15 000 endotoxin units (EU) ml(-1)] were present in the water. A statistically significant Spearman correlation coefficient of rho=0.94 between endotoxin (EU ml(-1)) and bacterial load (c.f.u. ml(-1)) was demonstrated. All of the aerosol samples contained detectable endotoxin. Further studies of the consequences of dental endotoxin exposure, and evaluation of means to prevent exposure, are warranted.

Fourteen miniature donkeys (Equus asinus) in a mid-Michigan herd of 38 animals presented with clinical signs of besnoitiosis, including the presence of typical tissue cysts in the ocular sclera, the buccal and nasal mucosa, together with characteristic dermatitis in specific areas of the body. The common histopathological change seen was the presence of many 100-200-microm diameter, thick walled, typical Besnoitia sp. tissue cysts together with a chronic cellular response associated with degenerating cysts. Microscopy of isolated scleral cysts and skin biopsies showed the presence of protozoal organisms consistent in morphology with that of Besnoitia bennetti bradyzoites. Molecular analysis of these parasites indicates that they differ from previously described coccidia, including Besnoitia sp., from rabbits and opossums. Isolated cases of infection with this agent have been reported infrequently in equids; however, this is the first report of an outbreak in a herd of donkeys in the United States.

A prospective study was carried out in an area of unstable malaria transmission in central Sudan to determine the efficacy and toxicity of quinine in pregnancy. Thirty-three pregnant women with severe Plasmodium falciparum malaria at mean 28.8 weeks gestational age were treated with quinine for 7 days. The mean body temperature on presentation for 3 patients who delivered prematurely was significantly higher than for those who delivered at term [39.2 +/- 0.7 degrees C versus 38.7 +/- 1.3 degrees C]. There were no significant difference between the 2 groups in other clinical or biochemical parameters. There were no clinically detectable congenital malformations and no auditory, visual or other neurological deficits in the babies at birth or 6 months later. Quinine may be safe in the treatment of severe falciparum malaria during pregnancy

The induction of pathological changes in Onchocerca volvulus infections is directly related to the presence of the microfilarial stage of this filarial nematode. Patients with either of the 2 major forms of the clinical disease (i.e., asymptomatic/mild [n=12] and severe [n=16] dermatopathology) were studied. The cellular immune responses (cell proliferation) of those with severe disease were stronger (stimulation index [SI], 12.3 +/- 1.9) than those with mild dermatopathological effects (SI, 2.9 +/- 0.6) or control patients (SI, 4.5 +/- 0.4). Cytoadherence antibody responses were greatest (grade 4) in the clinically severe group and only weak (grades less than or equal to1) in the mild group or the control patients. Ivermectin treatment was followed by an increase in immune responsiveness in those with initially poor responses. Thus, the degree of dermatopathological effect is related to the host's immune response against microfilariae, and ivermectin augments such responses.

Mectizan (Ivermectin) has been proved to be central to the control of onchoceriasis through self-sustainable community-based treatment. The possibility of parasitological unresponsiveness to this treatment or selection for drug resistance has emerged recently in many occasions. The reason for the reduced ability of Mectizan to maintain low levels of dermal microfilariae and early recurrent pruritus can only be speculated upon. Here, we report our own findings to address this particular issue.

Mass drug administration programs are the mainstay of current attempts to control and rid the globe of the two major filarial diseases of humans, onchocerciasis and lymphatic filariasis. There are a number of important components to developing and maintaining a treatment program; these include advocacy, communication, financial support, and the presence of local "champions" to lead the program. Two active country programs — lymphatic filariasis elimination in Tanzania and onchocerciasis control in Sudan — provide practical examples of these program needs. There are a number of positive consequences of a successful program that extend outside the primary goal of disease control or elimination; these include national infrastructure and personnel development. The challenges and consequent actions that could be taken to ensure the success of control programs are discussed.

Twenty years ago onchocerciasis was a disease generally ignored by the medical world, except by those who actually worked with the affected people in Africa and Latin America. Now, largely as a result of the success of mass vector control and drug treatment programs, this is a disease management model for developing countries. The recent literature on onchocerciasis has, not surprisingly, mainly focused on various aspects of control. Investigation into the more basic questions is needed to ensure continued effective disease control. The present mass drug control program is based on a single pharmaceutical, ivermectin (Mectizan), which acts almost exclusively on the microfilarial stage of the infection. Efforts are being made to identify other useful drugs; however, no major candidates have yet appeared. The identification of potential biochemical targets for anti-filarial compounds through a better understanding of the biochemistry of these worms is being pursued. The Onchocerca volvulus endosymbiont Wolbachia may provide a target for therapeutic intervention. An improved understanding of the genomics of O. volvulus has made possible the identification of strain differences in the parasites, and an appreciation of the relevance of these strain differences to the clinical disease, onchocerciasis. There is a need for a better understanding of the clinical disease, and the various pathogenic mechanisms that underly the different syndromes. It is particularly important to understand the pathological basis and mechanisms underlying the adverse responses that can occur with chemotherapy. Present control programs now need to be carefully monitored for effectiveness using new assessment tools, such as antigen assays and the identification of organisms in pools of vectors. Current efforts to control onchocerciasis must be coordinated with new chemotherapy-based control programs for other worm diseases that are emerging. The results of laboratory studies are increasingly being applied to improve the effectiveness of field-based control programs and their assessment. Such research is essential for progress towards the goals of controlling and eliminating onchocerciasis.

This paper reports evidence of members of the bombesin-like peptide family in a number of nematodes, including Caenorhabditis, Panagrellus, Dirofilaria, Onchocerca, Brugia, Haemonchus, Ostertagia, Toxocara, and Ascaris. One of these (Ostertagia) secretes the bombesin-like material into its environment. Specific binding of gastrin-releasing peptide to the hypodermis consistent with the presence of receptors was demonstrated. These data suggest that this class of peptides may play an important role in nematode hypodermal physiology.

The mixed leukocyte population obtained from the peritoneum of the August rat is a potentially important experimental model of inherent eosinophilia that has not been well characterized. In the present study, isolated cell preparations generated a concentration-dependent release of leukotriene (LT) C(4) when exposed to the Ca(2+) ionophore A23187, reaching maximal stimulation at 5.0 muM. This response was inhibited by the 5-lipoxygenase activating protein antagonist MK-886 (0.1 muM), nominally Ca(2+) and Mg(2+)-free incubation media and by activation of protein kinase C via phorbol 12-myristate 13-acetate (50 nM). These findings establish a model system for investigating LTC(4) profiles contingent with innate peritoneal eosinophilia and are consistent with the hypothesis that cellular LTC(4) biosynthesis is phosphoregulated.

Ivermectin efficacy and post-treatment reactions in asymmetric severe reactive ochodermatitis (sowda) were studied in 8 patients with sowda syndrome and 6 with mild generalized onchodermatitis in Sudan. Initial skin snips from 12 patients contained microfilariae (1-9 per mg skin). Patients were treated in hospital with a single oral dose of c. 150 mu g/kg ivermectin (103-200 mu g/kg) and monitored for frequency and severity of post-treatment reactions for 4 weeks. Serial samples of heparinized blood were collected over the first 24 h after treatment for determination of ivermectin pharmacokinetics. Skin snips from all patients on days 3 and 28 revealed no microfilariae. Post-treatment reactions were more common and severe in individuals with sowda; they consisted mainly of musculoskeletal pain, local swellings with pitting oedema, and lymph gland tenderness and enlargement. No relation was established between these reactions, the microfilarial infection intensity, or the plasma pharmacokinetic profiles. A single oral dose of ivermectin cleared the skin of microfilariae and led to improvement of symptoms and dermatological signs of sowda, but resulted in more marked reactions than in cases of generalized onchodermatitis.

In Ecuador, previous epidemiological studies of onchocerciasis affecting a population consisting of Blacks of African origin and Amerindians of the Chachi tribe, have demonstrated different frequencies of clinical disease manifestations in each racial group. To explore the possibility of differential antibody responses to O. volvulus antigens in this population, a subset of sera from these two groups of Ecuadorians with onchocerciasis was analyzed for antibody to O. volvulus antigens using ELISA and Western blot techniques. Significantly higher levels of immunoglobulin against O. volvulus were demonstrated in Chachi Indians compared to native Blacks of African origin: total Ig (p = 0.049), IgG (P = 0.002) IgG4 (p = 0.019) and IgA (p = 0.035). Western blot analysis of sera from 41 persons demonstrated more intense and more frequent recognition of low molecular weight parasite antigens in the Chachi compared to a similar group of male and female Ecuadorian Blacks. These data suggest that the intensity of antibody responses to O. volvulus antigens may reflect the influence of racial factors in the pathogenesis of onchocerciasis in Ecuador.

Although onchocerciasis is one of the most important diseases of the skin and eye in the tropical world, to date there has been no formal consensus regarding the description and terminology of skin lesions. Furthermore, the contribution of cutaneous pathology to the morbidity and socio-economic effects of the disease has been largely neglected. We present a clinical classification and grading system for recording the cutaneous changes of onchocerciasis, and propose that this system be used as a standard method of description to convey clinical information between workers in all endemic areas to assist local and comparative research.

Microbiological studies on dental unit water samples from 150 operatories revealed widespread and unacceptably high levels of contamination. Biofilm formation along the walls of the fine-bore waterlines is primarily responsible. These findings are reviewed and their relevance to contemporary infection control concerns discussed.