Immunodiagnosis of bancroftian filariasis—Problems and progress

The immunodiagnosis of bancroftian filariasis is a major challenge to the immunoparasitologist. Significant progress is yet to be made in developing convenient laboratory animal model and inin vitro cultivation of filarial parasites making it very difficult to obtain required amount of parasite material for research. Parasitological examination techniques are not useful in low microfilaraemia, occult or chronic.filarial infections. A precise and accurate immunodiagnostic technique is very much needed for successful filaria control programmes. Such a test will also avoid the need for laborious night blood examination in bancroftian filariasis. Due to comparatively easy availability, a good amount of work has been done to explore immunodiagnostic potential of heterologous filarial antigens isolated fromLitomosoide carinii, Dirofilaria immitis, Brugia malayi, Setaria digitata, Setaria cervi and number of other filarial species. However, there has been limited or no significant success due to number of false negative and false positive reactions. Extensive study has also been made with antigens isolated fromWuchereria bancrofti microfilariae. Soluble antigens of microfilariae have been used in different immunological techniques such as skin test, counter immuno electrophoresis, indirect haemagglutination test, indirect fluorescent antibody test and enzyme linked immunosorbent assay. Fractionation of Wuchereria bancrofti microfilarial soluble antigens yielded mfS3e antigen fraction which was found to be highly reactive in microfilaraemia by enzyme linked immunosorbent assay, but the yield of the purified antigen was not sufficient enough to make it a practical proposition for large scale isolation of antigen. Wuchereria bancrofti microfilarial excretory-secretory antigens were found to be specific and highly sensitive requiring as little as 0.35 ng antigen protein per well in penicillinase enzyme linked immunosorbent assay for detection of filarial antibody. One ml of culture fluid was found to be sufficient for 400,000 tests. Field evaluation of this test showed that it can replace laborious night blood examination. Assay systems have been developed for detection of filarial antigen in serum, urine, hydrocele fluid and immune complexes using immunoglobulins from chronic filarial sera and antisera to excretory filarial antigens. Further purification of excretory-secretory antigens by affinity chromatography and production of monoclonal antibodies should hopefully give suitable reagents for use in sensitive assays such as enzyme immuno assay and immuno radiometric assay, providing an ideal assay system for detection of active filarial infection in the not too distant future.     

Longitudinal follow‐up of malaria transmission dynamics in two villages in a Sahelian area of Niger during a nationwide insecticide‐treated bednet distribution programme

Malaria transmission was monitored in two villages in the Sahel zone of Niger over 4 years. During this period, a nationwide vector control programme was carried out in which insecticide‐treated bednets were distributed free to mothers of children aged <5 years. Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) were found to be the major malaria vectors. The dynamics of An. gambiae s.l. did not vary dramatically over the study period although the proportion of female mosquitoes found resting indoors decreased in both villages and, in one village, the parity rate and sporozoite index were significantly reduced after bednet distribution. By contrast with An. gambiae, the dynamics of Anopheles funestus altered greatly after the bednet distribution period, when adult density, endophagous rate and sporozoite rates decreased dramatically. Our observations highlight the importance of quantifying and monitoring the dynamics and infections of malaria vectors during large‐scale vector control interventions.     

Diagnostic utility of monoclonal antibodies raised against microfilarial excretory-secretory antigens in bancroftian filariasis

Two monoclonal antibodiesWuchereria bancrofti E 33 andWuchereria bancrofli E 34 raised againstWuchereria bancrofti microfilarial excretory-secretory antigens were studied for their diagnostic utility.Wuchereria bancrofti E 34 monoclonal antibody was found to be relatively specific and sensitive in detection of circulating filarial antigen. WhenWuchereria bancrofti E 34 monoclonal antibody was used alongwith immunoglobulin G fraction of human filarial serum immunoglobulins in double antibody sandwich enzyme linked immunosorbent assay. 68% of microfilaraemic sera (26 out of 38). 12% of clinical filarial sera (3 out of 25), 13% endemic normal sera (2 out of 15) and none of the 20 non-endemic normal sera showed the presence of filarial antigen. The filarial antigen detected byWuchereria bancrofti E 34 monoclonal antibody in double antibody sandwich enzyme linked immunosorbent assay is possibly associated with the active stage (microfilaraemia) of infection.     

Impact of permethrin-impregnated mosquito nets compared with DDT house-spraying against malaria transmission by Anopheles farauti and An.punctulatus in the Solomon Islands

Abstract. In villages of northern Guadalcanal in the Solomon Islands, where the predominant malaria vector is An.farauti No. 1 and An. puctulatus is also involved, malaria transmission rates were compared for three zones: (1) non-intervention: 438 people in seventeen villages; (2) residual DDT house-spraying two cycles per year: 644 people in thirty villages; (3) bednets impregnated with permethrin 0.5 g/m² twice per year, used by 580 people in sixteen villages. Regular DDT spraying in zones 1 and 3 had been withdrawn 18 months previously. Malariological blood smear surveys of children aged 1-9 years in August 1986 to January 1987 showed a mean-baseline malaria parasite rate of 38% (32/84). By February 19 88 , 18 months after introduction of impregnated bednets, the Plasmodium falciparum infection rate in children was lowest in the zone using impregnated bednets (21% of 29), intermediate in the untreated zone (29% of 34) and highest in the DDT zone (46% of 53), but these differences were not statistically significant. P.vivax infection rates were 9–14%. Using ELISA tests for malaria circumsporozoite antigen in the vectors, overall positivity rates were 0.7% of 49 ,902 An.farauti and 2.54% of 118 An.punctulatus, comprising 228 P.falciparum and 124 P. vivax infections. In the study zones, vector positivity rates were 0.93% of 31 ,615 An.farauti in the untreated zone; 0.32% of 16, 883 An.farauti in the DDT zone; 0.07% of 1404 An.farauti and 2.54% of 118 An.puctulatus in the impregnated bednet zone. There was no significant correlation between malaria parasite rates in the vectors and the children. Entomological inoculation rates were consistently highest in the untreated zone (1.6–2.8 infective bites/night), intermediate in the DDT zone (0.8– 1.1/night) and significantly lowest in the bednet zone (0.03-0.23/night). Geometric mean densities of P.falciparum sporozoites were also significantly higher in the DDT zone (50% > 10,000 sporozoites/mosquito compared with 20% in untreated zone). The highest individual infection density was an estimated 52,080 sporozoites of P.falciparum in a specimen of An.punctulatus from the bednet zone. P.vivax sporozoite densities were not significantly different between zones, and both species of vector had similar mean sporozoite loads for both species of malaria. It is concluded that permethrin-impregnated mosquito nets exerted significantly more impact on vector infectivity and the inoculation rate than resulted from DDT spraying. Even so, the inoculation rate for people in the bednet zone remained at one infective bite every 4–32 days, an insufficient reduction to control malaria without additional countermeasures. Ineffectiveness of house-spraying and the limited impact of impregnated bednets are attributed to exophily and other behavioural aspects of An. farauti.     

Behaviour of Anopheles albimanus in relation to pyrethroid-treated bednets

Abstract. Responses of the malaria vector Anopheles albimanus to pyrethroid impregnated bednets made of cotton or nylon, compared with untreated nets, were investigated in houses occupied by two people inside and/or outside two bednets, in coastal Chiapas, México. The pyrethroid used was lambdacyhalothrin 30mga.i./m². Bioassay mortality rates of An.albimanus exposed to treated nets for 3 or 15min, rose from 40–55% to 90–100% for nylon nets 3–19 weeks post-treatment, but were consistently lower for treated cotton nets. An.albimanus females (collected unfed on human bait) were released in houses surrounded by curtains for trapping mosquitoes that exited from the house. Floor sheets were used in and around each experimental house for retrieving any mosquitoes knocked-down and/or killed. During post-treatment assessment for 17 weeks, An.albimanus blood-feeding success rates were 23–24% with untreated nets, 14–18% with treated cotton nets and 8–15% with treated nylon nets, significantly reduced when both human baits were inside the treated bednets, but not when one or more baits were outside the treated bednet(s) within the house. Proportions of mosquitoes leaving houses <3 h post-release were 53–59% from houses with untreated bednets versus 65–78% with treated bednets. Except in one case (when both humans were outside treated cotton nets), these increased early exit rates were significant, whether or not the human baits were inside the treated bednets indoors. Mortality rates of An.albimanus females exiting overnight (22.00–06.00 hours) averaged 15–39% from houses with treated cotton and 16–46% with treated nylon nets, very significantly greater than the control mean rates of 6–8% mortality with untreated nets. Observations on wild-caught An. albimanus females marked with fluorescent powder and released indoors revealed that few mosquitoes (3–11 %) actually contacted the bednets unless both human baits remained under them - when contact rates were 22% on treated nylon, 23% on treated cotton and 42% on untreated nets (P= 0.05). The mean resting time was significantly longer on untreated (14.4min) than on treated nylon (5.8min) or cotton (9.5 min) bednets, whereas mean resting times on other surfaces indoors were 16.5–19.8min. Proportions exiting within 2h of release were significantly more from houses with treated houses (33–35%) than with untreated nets (8%). However, mortality rates of mosquitoes that landed on treated nets were very significantly greater (90–100%) than after landing on untreated nets (10%). Thus, despite some excito-repellency, lambdacyhalothrin-impregnated bednets (especially made of nylon) proved to be effective as an alternative to house-spraying against An.albimanus.     

Permethrin-impregnated bednets are more effective than DDT house-spraying to control malaria in Solomon Islands

Abstract. A field trial compared DDT house-spraying with permethrin-impregnated bednets for malaria control in Solomon Islands from 1987 to 1991. Mortality-rates of malaria vector Anopheles farauti in exit window traps were 11.6% from an untreated hut, 10.1% from a hut sprayed with DDT 2 g/m², and 98% of those from a hut in which the occupants used bednets treated with permethrin 0.5 g/m². Since bioassays of the DDT-sprayed walls (15 min exposure in W.H.O. standard test cones) gave 77% mortality of Anfarauti , it was concluded that the insignificant impact of DDT could be explained by the exophilic behaviour of endophagic vectors, whereas the greater impact of permethrin was attributed to the more effective exposure of Anfarauti females to the impregnated bednets - attracted by the occupants. The parous rate was higher indoors, except in the area with permethrin-impregnated bednets. It was therefore concluded that permethrin-impregnated bednets reduced the mean longevity of Anfarauti and hence its vectorial capacity. The circumsporozoite (CS) antigen positivity rate of Anfarauti in the DDT area was 0.18% outdoors, significantly less than 1.42% indoors. In the comparison area CS rates were 0.65% outdoors and 0.75% indoors. CS antigen was not detected in Anfarauti from the bednet area, indicating the apparent prevention of malaria transmission. As DDT spraying was so much less effective, it was discontinued in 1993 and permethrin-impregnated bednets are now the principal malaria control method in Solomon Islands.     

Immune response to an allergenic fraction ofSetaria digitata in human filariasis

A low molecular weight antigenic fraction has been isolated from saline-soluble extracts of cattle filarial parasiteSetaria digitata. This glycoprotein fraction (Fr III) which appears to have low phosphorylcholine content cross-reacted with infective larval (L₃) antigens ofWuchereria bancrofti. Binding of human chronic filarial serum with L₃ antigens could be inhibited partially by Fr III. The fraction elicited immediate cutaneous hypersensitivity reaction in people living inWuchereria bancrofti endemic regions. The proportion of skin test positive cases was found to be highest in endemic normals in contrast to infected cases. IgE levels were however not different in chronic filariasis and in endemic normals (or in asymptomatic microfilaraemic carriers). On the other hand, specific IgG level was considerably enhanced only in chronic filariasis     

Anopheles gambiae gonotrophic cycle duration, biting and exiting behaviour unaffected by permethrin-impregnated bednets in The Gambia

Abstract. Permethrin-impregnated bednets protect children against malaria in The Gambia, where Anopheles gambiae complex mosquitoes are the main vectors of malaria. However, no effect has been found on mosquito density, parous rates or sporozoite rates in An.gambiae sensu lato populations; only a reduction in the numbers of mosquitoes resting indoors in rooms with treated bednets. A possible explanation for this paradox is that exposure to treated bednets leads to changed vector behaviour such as a shift in biting time, a diversion to biting outdoors instead of indoors, to biting animals instead of humans, or to increased duration of the gonotrophic cycle. To investigate these possibilities, we observed the biting and exiting behaviour of An.gambiae in ten pairs of villages, in half of which the residents used permethrin-treated bednets. The possible influence of treated bednets on the gonotrophic cycle length was evaluated by mark-release-recapture experiment. No significant difference was found between villages with treated and untreated bednets in the indoor/outdoor ratio of human biting, in mean biting times or in human blood indices of An.gambiae females found resting indoors in the mornings. The proportions of unfed, fed or gravid An.gambiae females collected in exit traps, and the number of females exiting showed no significant differences between rooms with treated and untreated bednets. Indications for a gonotrophic cycle length of 2 days were found. No evidence for any change in duration of the gonotrophic cycle in relation to exposure to treated bednets was found, although the number of recaptures was low in the villages with treated bednets. Since equal numbers of infective An.gambiae were found in villages with treated or untreated bednets, and no changes in mosquito behaviour were detected, we cannot account for how children are protected against malaria by treated bednets. One possibility is that mosquitoes divert to bite other hosts, including adults.     

Population genomics of the filarial nematode parasite Wuchereria bancrofti from mosquitoes

Wuchereria bancrofti is a parasitic nematode and the primary cause of lymphatic filariasis – a disease specific to humans. W. bancrofti currently infects over 90 million people throughout the tropics and has been acknowledged by the world health organization as a vulnerable parasite. Current research has focused primarily on the clinical manifestations of disease and little is known about the evolutionary history of W. bancrofti. To improve upon knowledge of the evolutionary history of W. bancrofti, we whole genome sequenced 13 W. bancrofti larvae. We circumvent many of the difficulties of multiple infections by sampling larvae directly from mosquitoes that were experimentally inoculated with infected blood. To begin, we used whole genome data to reconstruct the historical population size. Our results support a history of fluctuating population sizes that can be correlated with human migration and fluctuating mosquito abundances. Next, we reconstructed the putative pedigree of W. bancrofti worms within an infection using the kinship coefficient. We deduced that there are full‐sib and half‐sib relationships residing within the same larval cohort. Through combined analysis of the mitochondrial and nuclear genomes we concluded that this is likely a results of polyandrous mating, the first time reported for W. bancrofti. Lastly, we scanned the genomes for signatures of natural selection. Annotation of putative selected regions identified proteins that may have aided in a parasitic life style or may have evolved to protect against current drug treatments. We discuss our results in the greater context of understanding the biology of an animal with a unique life history and ecology.     

Evaluation of fractionatedWuchereria bancrofti microfilarial excretory-secretory antigens for diagnosis of bancroftian filariasis by enzyme linked immunosorbent assay

TheWuchereria bancrofti microfilarial excretory-secretory antigens were fractionated into ES1, ES2, ES3 and ES4 by ultra-membrane filtration and evaluated for their diagnostic utility by enzyme linked immunosorbent assay. Three of the four fractions showed antigenic activity (ES2, ES3 and ES4). The antigen fractions ES2 and ES4 were highly active in the detection of filarial IgM antibody in clinical filariasis and microfilaraemia respectively. The chemical characterization of the ES2 and ES4 antigen fractions showed that they were glycoproteins     

Permethrin-impregnated bednet effects on resting and feeding behaviour of lymphatic filariasis vector mosquitoes in Kenya

The impact of permethrin-impregnated bednets on resting and feeding behaviour of mosquito vectors of Wuchereria bancrofti, causing human lymphatic filariasis was studied in six pairs of villages (treated and untreated) before and after intervention. The study villages were in Kwale District, near the coast of Kenya, where Bancroftian filariasis is highly endemic, transmitted by a combination of both anopheline and culicine mosquito vectors. Mosquitoes were collected weekly in each village, indoors (using pyrethrum spray catches) and outdoors (using pit traps) during 3–4 months following the long rainy season. Of the filariasis vector species of mosquitoes collected in 1994 before intervention, 33.6% were members of the Anopheles gambiae complex, 30% were An. funestus and 36.4% were Culex quinquefasciatus. PCR analysis of the An. gambiae complex species collected in 1995 demonstrated that 98.5% were An. gambiae sensu stricto, 1% An. arabiensis and 0.5% An. merus. Introduction of impregnated bednets in 1995 significantly reduced the number of indoor-resting An. gambiae s.l. by 94.6% and An. funestus by 96.7%, but there was no change in the number of Cx quinquefasciatus collected indoors. The number of outdoor-resting An. gambiae s.l. was significantly reduced, whereas densities of An. funestus and Cx quinquefasciatus remained unaffected outdoors. ELISA analysis of mosquito bloodmeals showed a shift from human to animal feeding after the introduction of treated nets. The human blood index (HBI) for indoor resting Cx quinquefasciatus was reduced from 93.1% to 14.4%. Vector potential based on the HBI and mosquito density was estimated to be reduced by 99% for An. gambiae s.l., 98% for An. funestus and 97% for Cx quinquefasciatus and vectorial capacity would be suppressed even more by the impact on the vector survival rates (not measured). These results suggest that permethrin-impregnated bednets give effective personal protection against transmission of W. bancrofti by An. gambiae, An. funestus and Cx quinquefasciatus in East Africa.     

Effects of community-wide use of lambdacyhalothrin-impregnated bednets on malaria vectors in rural Sierra Leone

Abstract. The effect of community-wide use of bednets treated with lambdacyhalothrin 10mg/m² on the malaria vector Anopheles gambiae (forest form) was evaluated in Sierra Leone. Sixteen similar villages near the town of Bo were randomly allocated either to remain without nets or to receive treated bednets for all inhabitants, with effect from June 1992. Mosquitoes were sampled using human biting catches on verandas, light-trap catch (beside an occupied untreated bednet), window exit-trap catch and pyrethrum spray collections. During the first year of intervention (June 1992 to July 1993) the treated bednets provided personal protection for people sleeping under them, but had very little impact on densities of An. gambiae collected on human bait. The human blood index (HBI) of An.gambiae was not affected (HBI = 99% in villages with and without nets). An.gambiae parous rates were significantly reduced in all intervention villages, but malaria sporozoite rates fell in only some of the villages. These results are intermediate between those obtained from other projects in Tanzania and Burkina Faso, where treated bednets reduced man-biting, parity and sporozoite rates, versus The Gambia where treated bednets had no significant impact on any of these factors. Possible reasons for these contrasted findings are discussed.     

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate,Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.     

Genetic epidemiology of lymphatic filariasis in American Samoa after mass drug administration

Over 892 million people in 48 countries are at risk of infection by nematodes that cause lymphatic filariasis. As part of the Global Programme to Eliminate Lymphatic Filariasis, mass drug administration is distributed to communities until surveillance indicates infection rates are below target prevalence thresholds. In some countries, including American Samoa, lymphatic filariasis transmission persists despite years of mass drug administration and/or has resurged after cessation. Nothing is known about the population genetics of Wuchereria bancrofti worms in Polynesia, or whether local transmission is persisting and/or increasing due to inadequate mass drug administration coverage, expansion from residual hotspots, reintroduction from elsewhere, or a combination. We extracted DNA from microfilariae on blood slides collected during prevalence surveys in 2014 and 2016, comprising 31 pools of five microfilariae from 22 persons living in eight villages. We sequenced 1104 bp across three mitochondrial markers (ND4, COI, CYTB). We quantified parasite genetic differentiation using variant calls and estimated haplotypes using principal components analysis, F-statistics, and haplotype networks. Of the variants called, all but eight were shared across the main island of Tutuila, and three of those were from a previously described hotspot village, Fagali’i. Genotypic data did not support population genetic structure among regions or villages in 2016, although differences were observed between worms collected in Fagali’i in 2014 and those from 2016. Because estimated haplotype frequency varied between villages, these statistics suggested genetic differentiation, but were not consistent among villages. Finally, haplotype networks demonstrated American Samoan sequence clusters were related to previously published sequences from Papua New Guinea. These are, to our knowledge, the first reports of W. bancrofti genetic variation in Polynesia. The resurgent parasites circulating on the main island of American Samoa represent a single population. This study is the first step towards investigating how parasite population structure might inform strategies to manage resurgence and elimination of lymphatic filariasis.     

Seasonal abundance and biting behaviour of Anopheles punctulatus and An.koliensis in Malaita Province, Solomon Islands, and a trial of permethrin impregnated bednets against malaria transmission

Seasonal abundance of the malaria vectors Anopheles punctulatus Donitz and An.koliensis Owen in Bilimanu, an isolated inland village with forty-two houses in Malaita Province of the Solomon Islands, was monitored over 28 months by means of all-night landing/biting catches at one site during June 1985 to September 1987. Totals of 1250 An.punctulatus and 141 An.koliensis were collected, the latter being the largest number of this species ever caught at any locality in the Solomons. Bednets impregnated with permethrin 0.5 g/m² were introduced in December 1986 to be used at night by all 190 villagers for protection against malaria vectors. Bioassay tests with An.punctulatus blood-fed females exposed under nets for lOmin resulted in 100% mortality up to 50 weeks post-impregnation. For An.punctulatus, the main vector species, the mean catch (indoors + outdoors) per man hour was 2.9 (range 0.7–13.2) before a cyclone on 19 May 1986, and, 0.66 (0.2-2.7) after the cyclone. The vector survival rates were usually high before the cyclone, but erratically lower thereafter for An.punctulatus. An.koliensis disappeared after the cyclone. Both An.punctulatus and An.koliensis consistently showed higher rates of biting man indoors than outdoors and their diel biting cycle showed a peak around midnight. Outdoors, the parous proportion of An.punctulatus was twice the nulliparous, and nearly so indoors. Following intervention with permethrin-treated bednets, the mean catch of An.punctulatus fell to 0.35 per man-hour (monthly range 0–1.5). The Plasmodium falciparum malaria infection rate reduced from 10% pre-intervention to zero in September 1987, 9 months after intervention, and then rose again. With re-impregnation of bednets in November 1987, the P.falciparum infection rate was again reduced by March 1988. No effect of treated bednets was observed on the prevalence of P.vivax malaria.     

Filarial worms reduce Plasmodium infectivity in mosquitoes

Background

Co-occurrence of malaria and filarial worm parasites has been reported, but little is known about the interaction between filarial worm and malaria parasites with the same Anopheles vector. Herein, we present data evaluating the interaction between Wuchereria bancrofti and Anopheles punctulatus in Papua New Guinea (PNG). Our field studies in PNG demonstrated that An. punctulatus utilizes the melanization immune response as a natural mechanism of filarial worm resistance against invading W. bancrofti microfilariae. We then conducted laboratory studies utilizing the mosquitoes Armigeres subalbatus and Aedes aegypti and the parasites Brugia malayi, Brugia pahangi, Dirofilaria immitis, and Plasmodium gallinaceum to evaluate the hypothesis that immune activation and/or development by filarial worms negatively impact Plasmodium development in co-infected mosquitoes. Ar. subalbatus used in this study are natural vectors of P. gallinaceum and B. pahangi and they are naturally refractory to B. malayi (melanization-based refractoriness).

Methodology/Principal Findings

Mosquitoes were dissected and Plasmodium development was analyzed six days after blood feeding on either P. gallinaceum alone or after taking a bloodmeal containing both P. gallinaceum and B. malayi or a bloodmeal containing both P. gallinaceum and B. pahangi. There was a significant reduction in the prevalence and mean intensity of Plasmodium infections in two species of mosquito that had dual infections as compared to those mosquitoes that were infected with Plasmodium alone, and was independent of whether the mosquito had a melanization immune response to the filarial worm or not. However, there was no reduction in Plasmodium development when filarial worms were present in the bloodmeal (D. immitis) but midgut penetration was absent, suggesting that factors associated with penetration of the midgut by filarial worms likely are responsible for the observed reduction in malaria parasite infections.

Conclusions/Significance

These results could have an impact on vector infection and transmission dynamics in areas where Anopheles transmit both parasites, i.e., the elimination of filarial worms in a co-endemic locale could enhance malaria transmission.     

Evidence for <Emphasis Type="Italic">Wolbachia</Emphasis> symbiosis in microfilariae of <Emphasis Type="Italic">Wuchereria bancrofti</Emphasis> from West Bengal,India

Wolbachia are symbiotic endobacteria that infect the majority of filarial nematodes, including Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus. Recent studies have suggested that Wolbachia are necessary for the reproduction and survival of filarial nematodes and have highlighted the use of antibiotic therapy such as tetracycline/doxycycline as a novel method of treatment for infections caused by these organisms. Before such therapy is conceived and implemented on a large scale, it is necessary to assess the prevalence of the endosymbiont in W. bancrofti from different geographical locations. We present data from molecular and electron microscopic studies to provide evidence for Wolbachia symbiosis in W. bancrofti microfilariae collected from two districts (Bankura and Birbhum) of West Bengal, India.     

The influence of permethrin-impregnated bednets and mass drug administration on the incidence of Plasmodium falciparum malaria in children in Sabah, Malaysia

A small-scale trial was carried out in the Upper Kinabatangan district of Sabah, Malaysia, to determine the effect of using permethrin-impregnated bednets on malaria transmission. A total of 306 nylon bednets with cotton borders, impregnated at a dose estimated to have been 0.062 g permethrin/m2 of nylon netting, were distributed to 139 households in five villages. At the time of distributing bednets, mass drug administration with Fansidar plus primaquine was also administered to the human population to clear all parasitaemias due to Plasmodium falciparum Welch. In another village, for comparison, mass drug administration was the only intervention. After intervention measures in December 1984 and January 1985, the parasite rates in children declined in all villages during the first month, significantly more in the villages with impregnated bednets than in the control, thus proving that the nets had an impact on malaria. However, after about 2 months, parasite rates started to increase again. After 4-6 months, parasite rates in the villages with bednets approached the rate in the control village without nets. The increase in parasite rates was paralleled by a significant deterioration in the quality, physical condition and the degree of non-utilization of bednets. Entomological evaluation proved the efficacy of permethrin-impregnated nets for controlling Anopheles balabacensis Baisas and other anophelines. Bioassays (1 h exposure) of permethrin-impregnated bednets gave 100% mortality initially and 44-61% mortality after 85-106 days. Mosquito collections in treated bednets were significantly reduced for at least 217 days. The project failed to achieve prolonged suppression of malaria transmission for a combination of entomological, sociological and practical reasons which are discussed in relation to the objectives and implementation of future bednet studies.     

Step towards elimination of Wuchereria bancrofti in Southwest Tanzania 10 years after mass drug administration with Albendazole and Ivermectin

BackgroundLymphatic filariasis is a mosquito transmitted parasitic infection in tropical regions. Annual mass treatment with ivermectin and albendazole is used for transmission control of Wuchereria bancrofti, the infective agent of lymphatic filariasis in many African countries, including Tanzania.MethodologyIn a general population study in Southwest Tanzania, individuals were tested for circulating filarial antigen, an indicator of W. bancrofti adult worm burden in 2009 before mass drug administration commenced in that area. Seven annual rounds with ivermectin and albendazole were given between 2009 and 2015 with a population coverage of over 70%. Participants of the previous study took part in a follow-up activity in 2019 to measure the effect of this governmental activity.FindingsOne thousand two hundred and ninety nine inhabitants of Kyela district in Southwest Tanzania aged 14 to 65 years who had participated in the study activities in 2009 were revisited in 2010/11 and 2019. Among this group, the prevalence of lymphatic filariasis of the 14–65 years olds in 2009 was 35.1%. A follow-up evaluation in 2010/11 had shown a reduction to 27.7%. In 2019, after 7 years of annual treatment and an additional three years of surveillance, the prevalence had dropped to 1.7%, demonstrating successful treatment by the national control programme. Risk factors for W. bancrofti-infection were the occupation as farmer, male sex, and older age. Most infected individuals in the 2019 follow-up study already had a positive test for filarial antigen in 2009 and/or 2010/11.ConclusionsThis data supports the findings of the Tanzanian Neglected Tropical Disease Control Programme (NTDCP), who conducted Transmission Assessment Surveys and found an impressive reduction in the prevalence of LF in children. Our results complement this data by showing a similar decrease in prevalence of LF in the adult population in the same area. The elimination of LF seems achievable in the near future.