Mark L. Eberhard

MATERNAL FILARIAL INFECTION AS RISK FACTOR FOR INFECTION IN CHILDREN

Familial clustering of filarial infection was investigated through random house-to-house surveys of 643 individuals in Leogane, Haiti, an area with endemic Bancroftian filariasis. Children of infected mothers were 2.4 to 2.9 times more likely to be infected than were those of amicrofilaraemic mothers. Filarial-specific cellular responsiveness in amicrofilaraemic children born to infected mothers was lower than that in amicrofilaraemic children born to amicrofilaraemic mothers. No effect of paternal infection status was seen. The findings show that maternal infection is a risk factor for filarial infection in children and is associated with altered parasite-specific immune reactivity.

Ocular Onchocerca infections in two dogs in western United States

Two dogs, one from California and one from Arizona, were found to have aberrant infections caused by filarial nematodes of the genus Onchocerca. In both cases, the parasites are localized in or near the eye. In one case the worm was located in the cornea and was surgically removed. In the second case, a very marked granulomatous reaction was induced in the retrobulbar space, mimicking an abscess. This eye was enucleated. The worms in both instances were female, and were gravid, i.e. contained microfilariae in utero, indicating that a male worm(s) had been present and mating had occurred. The exact identity of the species of Onchocerca responsible cannot be determined, although the features observed are most like Onchocerca lienalis of cattle. These cases represent the fourth and fifth such cases reported from the US, and are especially interesting because of the unusual location of the worms, the small number of recognized cases, and the similarity to a recent zoonotic human infection.

Molecular and morphologic characterization of a Cryptosporidium genotype identified in lemurs

This study reports the molecular and morphologic characterization of a Cryptosporidium sp., identified in stools of captive lemurs Propithecus verreauxi coquereli. Stool samples were collected from seven animals (n=7) presenting episodes of diarrhea. Bright-field light microscopy of stool smears stained with modified acid-fast technique revealed the presence of Cryptosporidium sp. oocysts in four of the stool samples analyzed. All microscopically positive samples were confirmed by PCR using primers designed to amplify DNA fragments from two independent loci, i.e. the Cryptosporidium oocyst wall protein (COWP) gene and the small subunit ribosomal RNA (ssrRNA) gene. Phylogenetic analysis based on the full-length ssrRNA gene placed this isolate within a clade that contains all currently known C. parvum species/genotypes, closely related to the C. parvum pig genotype. Comparison with partial ssrRNA sequences available in the GenBank revealed 100% sequence identity with the genotype previously identified in Canadian patients. This finding was confirmed further by comparison of the COWP gene partial sequences.

Zoonotic Onchocerca (nematoda:filarioidea) in the cornea of a colorado resident

OBJECTIVEnA female patient, resident in the state of Colorado, presented with iritis of the right eye. Slit-lamp examination showed the presence of a thin, threadlike worm entwined in the cornea. The patient was taken to surgery for removal of the parasite.nnnDESIGNnA case report.nnnINTERVENTIONnA 3-mm-long supertemporal incision was made in the cornea and further dissected until the worm could be grasped and removed by gentle traction.nnnRESULTSnThe worm, a filarial nematode, was identified as a member of the genus Onchocerca, most likely Onchocerca cervicalis, a natural parasite of horses. The patient had an uneventful recovery, and 1 week after surgery, her visual acuity, intraocular pressure, and corneal edema were all resolving.nnnCONCLUSIONnIn the United States and elsewhere, most cases of zoonotic filarial infection involving the eye are caused by Dirofilaria or Dipetalonema-like worms. However, the current case was caused by a species of Onchocerca. This is the first case of zoonotic Onchocerca from the eye to be reported, only the second case of zoonotic Onchocerca in the United States, and the seventh case worldwide. The worm was removed surgically, and the patient had an uneventful recovery.

Gongylonema Infection of the Mouth in a Resident of Cambridge, Massachusetts

We report a case of Gongylonema infection of the mouth, which caused a migrating, serpiginous tract in a resident of Massachusetts. This foodborne infection, which is acquired through accidental ingestion of an infected insect, such as a beetle or a roach, represents the 11th such case reported in the United States.

Long-term suppression of microfilaraemia following ivermectin treatment.

Lymphatic filariasis has been difficult to control until recently because of the lack of a suitable drug for treatment. Ivermectin has proven safe and effective at reducing levels of circulating microfilariae. However, the apparent need to administer the drug every 6 to 9 months to keep microfilaraemia levels sufficiently suppressed to reduce transmission has been a major drawback to using ivermectin in community-based intervention programmes. In a study conducted in Haiti, we have found that high doses of ivermectin suppress microfilaraemia levels for 2 years. Our findings suggest that a single dose of ivermectin can reduce transmission of lymphatic filariasis for extended periods of time, thus eliminating the need for costly biannual treatment.

Differentiating Dracunculus medinensis from D. insignis, by the sequence analysis of the 18S rRNA gene.

Abstract This study, undertaken as a component of the global Dracunculiasis Eradication Program (DEP), was designed to provide molecular tools to distinguish Dracunculus medinensis, the nematode causing human dracunculiasis, from other tissue-dwelling nematodes, including other Dracunculus species that infect humans and other animals. DNA was extracted from D. medinensis and from a closely related species that infects North American carnivores, D. insignis, so that the genes coding for the small-subunit ribosomal RNA (18S rRNA) of the parasites could be amplified, sequenced and compared. Sequences were obtained for 20 specimens of D. medinensis (from humans in Pakistan, Yemen and six African countries endemic for dracunculiasis) and three of D. insignis (from raccoons trapped in the state of Georgia in the southern U.S.A.). All of the D. medinensis 18S-rRNA sequences were found to be 1819 bases long and identical. The three D. insignis 18S-rRNA sequences were also found to be identical to each other but were 1821 bases long and differed from the D. medinensis 18S- rRNA sequence at eight positions (representing a difference of 0.44%). The 18S-rRNA coding region of a Guinea worm extracted from a dog in Ghana was indistinguishable from that of the D. medinensis isolates from human cases. These results provide the basis for the molecular differentiation of D. medinensis that will permit the DEP to determine, rapidly and accurately, whether a worm recovered from an area considered dracunculiasis-free is a specimen of D. medinensis or not.

Acquisition and expression of humoral reactivity to antigens of infective stages of filarial larvae

Measurement of anti‐larval responses in filaria‐exposed populations may shed light on the natural history of exposure to Wuchereria bancrofti. Using serum samples obtained by a cross‐sectional survey of 172 individuals from two neighbourhoods in Leogane, Haiti, antibody responses directed against infective stage filarial larvae (L3) were assayed by enzyme‐linked immunosorbent assay (ELISA), immunofluorescence (IFA), and immunoblot for the presence of anti‐larval antibodies. ELISA results indicated that virtually all members of both neighbourhoods mounted an anti‐larval antibody response within the first five years of life, suggesting that exposure to infection is universal. In a multiple linear regression analysis that modelled antibody levels as a function of age, gender, microfilaria status, and neighbourhood (as a proxy for transmission intensity), isotype‐specific antibody levels were found to be significantly influenced by both age and neighbourhood. Antibodies directed against the surface of L3 also were age‐dependent; the prevalence of IgG antibodies detected by IF A was significantly higher in children than in adults. The prevalence of antibody recognition of 16–7 and 72–3kDa L3 antigens on immunoblots was significantly greater for serum samples from microfilaraemic than amicrofilaraemic persons. These results suggest that antibody responses to larval antigens are influenced to varying degrees by age, transmission intensity, and microfilaremia status.

Investigation of the influence of maternal infection with Wuchereria bancrofti on the humoral and cellular responses of neonates to filarial antigens

Epidemiological data indicate that maternal filarial infection might be associated with increased susceptibility to filarial infection in offspring. To examine the influence of maternal infection on development of antifilarial immunity in neonates, paired cord and maternal sera and mononuclear cells were collected in an area where Wuchereria bancrofti infection is endemic. Anti-filarial humoral responses (IgG, IgM and IgE) non-parasite-specific humoral responses (total IgE), proliferation induced by filarial antigen and production of cytokines (interleukin-2, interleukin-4 and interferon-gamma) were all monitored. Few cord serum samples had detectable antifilarial IgM of IgE and neither these responses nor total IgE levels differed as a function of maternal infection status. Of cord-blood mononuclear cells assayed, a relatively small proportion exhibited reactivity to filarial antigens. Based on these limited responses to filarial antigens, few neonates display evidence of in-utero sensitization to filarial antigens.

Chemoprophylactic drug trials for treatment of dracunculiasis using the Dracunculus insignis-ferret model.

Groups of ferrets inoculated with Dracunculus insignis were treated with various anthelminthic compounds to evaluate the potential use of drugs in controlling the human parasite D. medinensis. The three primary compounds tested were diethylcarbamazine (DEC), albendazole (ALBZ), and invermectin (IVER); they were administered in dosages of 60 mg/kg, 10 mg/kg, and 1 mg/kg, respectively. Groups of animals received treatment either once at 60 days after inoculation, once at 60 and 90 days, or for 3 days at either 60 or 90 days postinoculation. The most marked decrease occurred in the animals that received treatment once at 60 and 90 days after inoculation. This was observed for all three drugs tested. Increased dosages, i.e., 3 days of treatment at 60 or 90 days did not result in decreased worm burdens. In no group was there a statistically significant reduction in worm burden when compared with controls. Two other compounds, metrifonate and CGP 6140, were tested in a more limited manner, but again the worm recovery rates were comparable with those in control groups. It would appear that existing drugs commonly used to treat helminthic infections are poor candidates for use in the campaign to eradicate guinea worm disease.