Effects of combined diethylcarbamazine and albendazole treatment of bancroftian filariasis on parasite uptake and development in Culex pipiens L

We studied effects of combined diethylcarbamazine (DEC) and albendazole (ALB) treatment on Wuchereria bancrofti microfilaria (MF) uptake and development of infective larvae (L3) in Culex pipiens. Consenting Egyptian adults with microfilaremia (MF > 300/mL) were treated with one or seven daily doses of DEC/ALB. Laboratory-reared mosquitoes were fed on subjects before and after treatment. MF uptake and infectivity (assessed by mosquito dissection) were reduced by 89.6% and 82.9%, respectively, 12 months after single-dose treatment and by 96.2% and 99.7%, respectively, after multi-dose treatment. The L3:mosquito ratio decreased by 88% to 0.082 after single-dose treatment and by 99.8% to 0.001 after multi-dose treatment. If high coverage rates can be achieved for several annual cycles, mass drug administration (MDA) with DEC/ALB has the potential to decrease transmission to unsustainable levels and eliminate filariasis in populations. Multi-dose MDA (especially in the first year) might interrupt transmission with fewer cycles than single-dose treatment.     

Uptake and development of Wuchereria bancrofti in Culex quinquefasciatus that fed on Haitian carriers with different microfilaria densities

In studies conducted between 1984 and 1986, the vector competency of Culex quinquefasciatus was assessed after bloodfeeding on 61 Haitian volunteers with different densities of Wuchereria bancrofti microfilariae (mf) and on 11 that were amicrofilaremic. Infected volunteers included persons previously given diethylcarbamazine citrate for 12 consecutive days and some that were untreated. Mosquitoes, derived from field-collected larvae, were released under bed nets and fed upon the volunteers while they slept. The mean mf uptake in mosquitoes that fed on 21 carriers with low to ultralow densities (1-28 mf/ml blood) was 0.2-4 mf/mosquito (mean = 1.7 mf). The observed infectivity rate of greater than 3,000 mosquitoes that fed on high (56-7,500 mf/ml blood; median = 525), low (11-49 mf/ml blood; median = 20), or ultralow (1-10 mf/ml blood; median = 3) density carriers was 4, 11, and 30 times higher, respectively, than the expected rate calculated from the estimated volume of the imbibed bloodmeal. These results indicate that "hidden" carriers (less than 50 mf/ml blood) may serve as a source of infection for mosquitoes, and support the increasing evidence that mosquitoes ingest more microfilariae than expected.     

Vector competence, for Wuchereria bancrofti, of the Anopheles populations in the Bongo district of Ghana

The ability of vector mosquitoes to transmit the microfilariae (mff) of Wuchereria bancrofti, especially when the levels of microfilaraemia in the humans on which the mosquitoes are feeding are very low, is very important for understanding the transmission dynamics of lymphatic filariasis. Data on the correlation between vector competence and the microfilarial load in the human host are also relevant to those trying to improve transmission models for this disease. The majority of the relevant studies have involved culicine rather than anopheline vectors. The competence of populations of Anopheles mosquitoes to transmit W. bancrofti in a district in the Upper East region of Ghana has now been investigated. The wild mosquitoes that fed on 20 volunteers under natural conditions were collected hourly during the night, from 21.00 hours on one day to 06.00 hours on the next. Overall, 1348 fed female mosquitoes--665 Anopheles, 662 Culex and 21 Aedes--were collected. Approximately 50% of the mosquitoes caught were killed immediately post-collection and dissected so that the number of W. bancrofti mff each had ingested could be counted. The remaining mosquitoes where dissected when they died (if this was within 12 days of collection) or when they were killed on day 12 post-collection. With the exception of one Culex mosquito that harboured one microfilaria, none of the Culex and Aedes mosquitoes were found infected with W. bancrofti. All of the other mosquitoes found infected were An. gambiae s.l. or An. funestus. When fingerprick samples of blood, collected hourly from the volunteers during the mosquito infection, were used to estimate the microfilaraemias in the blood on which these mosquitoes had fed, microfilarial uptake and the number of developing larvae were found to vary considerably even when the microfilaraemias in the bloodmeal source were similar. The results of a regression analysis on the pooled data for the Anopheles mosquitoes indicated the process of limitation, although larger samples need to be investigated to determine whether this process occurs only in An. gambiae s.l. or An. funestus or in both of these taxa.     

The impact of single-dose diethylcarbamazine treatment of bancroftian filariasis in a low-endemicity setting in Egypt

This study was designed to evaluate the effect of a single dose of diethylcarbamazine (DEC, 6 mg/kg) on Wuchereria bancrofti infections in a low-endemicity setting in Egypt (microfilaremia, or MF, 3.7%, median MF 34/mL). Subjects with MF or filarial antigenemia were treated and restudied 1 year later. Treatment with DEC dramatically reduced blood MF counts, with clearance in 69% of subjects. Treatment also reduced filarial antigen levels, but low clearance rates suggest that some adult worms survived treatment in most patients. Mass treatment was administered in one village; 27 months later, MF prevalence had decreased 84% (from 4.9% to 0.8%). These results show that single-dose DEC treatment can have a major effect on MF prevalence rates and levels in low-endemicity settings. Although the World Health Organization advocates repeated multidrug regimens for filariasis elimination, mass treatment with DEC alone may be sufficient to interrupt transmission in areas with low infection intensities and prevalence rates.     

A new realistic index of experimental transmission efficiency for Bancroftian filariasis

An index for experimental transmission of filariasis infection was first introduced by Kartman and subsequently modified by Wharton who defined it as 'the number of infective larvae per mosquito when the microfilaria count is 1.0 mm-3 of human blood'. Since mosquitoes may not imbibe the expected number of microfilariae from the donor's blood during feeding because of various interference phenomena and/or mechanisms, this paper presents the index mosquito meal (Index MM), based on the actual number of viable microfilariae that are observed in the mosquito gut immediately after feeding. The Index MM is defined as 'the number of infective larvae per mosquito when the microfilaria count is 1.0 mm-3 of mosquito blood meal'. The Index MM proved to be a more valid indicator of transmission potential than the Wharton Index when applied to the data of two controlled field studies where groups of laboratory-reared Culex pipiens molestus, the primary vector of filariasis in Egypt, were fed on volunteers having different microfilaraemic densities.     

Limitation and facilitation in the vectors and other aspects of the dynamics of filarial transmission: the need for vector control against Anopheles-transmitted filariasis

In certain filaria-mosquito combinations, the number of infective, third-stage larvae (L(3)) that develop in a mosquito is not proportional to the number of microfilariae (mff) ingested by that mosquito. As the number of mff ingested increases, the yield of L(3) per microfilaria may either increase (in a process known as 'facilitation') or decrease (in a process known as 'limitation'). Each ingested microfilaria that is successful (in terms of reaching the haemocoel) increases (facilitation) or decreases (limitation) the 'permeability' of the stomach wall for the next microfilaria. Limitation is seen in some culicine mosquitoes, especially the Aedes spp. that transmit Wuchereria bancrofti, which, in consequence, become relatively more efficient as vectors as they ingest fewer mff. This phenomenon makes the interruption of filarial transmission by Aedes spp. particularly difficult. As the survival of anopheline mosquitoes is adversely affected by filarial infection, the use of mass drug administrations (MDA) to reduce the prevalence and intensity of microfilaraemias may increase the mean lifespan of some of the local Anopheles species. If these same species also act as vectors of malarial parasites, effective, drug-based control of W. bancrofti may worsen the problem posed by malaria. Therefore, wherever malaria and bancroftian filariasis are co-endemic and caused by parasites transmitted by the same species of mosquito, MDA should be augmented by interventions (use of bednets or house-spraying) against adult Anopheles.     

A survey of bancroftian filariasis among South-East Asian expatriate workers in Saudi Arabia

In a survey of bancroftian filariasis among expatriate workers from five South-East Asian countries (India, Bangladesh, Sri Lanka, Thailand and the Philippines) where human filariasis is endemic, 762 individuals were examined in the Abha area (Asir) of south-western Saudi Arabia. A prevalence of microfilaraemia of 3.5% and a mean mf density of 6.0/20 mm³of blood was found among 259 Indian males only. In three out of 9 microfilaraemic cases, clinical signs suggestive of filariasis were observed. The only species identified was Wuchereria bancrofti showing strict nocturnal periodicity. Preliminary laboratory studies on the susceptibility of local mosquitoes to infection with W. bancrofti in which laboratory-bred Culex (C.) pipiens was fed directly on a microfilaraemic volunteer from Madras, South India, revealed that this species was highly susceptible to the Madras strain of the parasite with an average infection rate of 57 (range 41–75)% and a worm burden of 3.5 L³/infective mosquito. This is the first report that local Cx. pipiens mosquitoes may act as a potential vector of introduced bancroftian filariasis in Saudi Arabia. The potential danger of bancroftian filariasis importation and, more importantly, the establishment of new self-sustained foci of the disease is likely to depend upon the abundance of mf carriers and density of vector mosquitoes feeding on carriers.     

A field survey on the role of low-density microfilaraemia cases in the transmission of filariasis

In former filariasis endemic areas, where the disease has been basically controlled, a few cases of low-density microfilaraemia remain. A survey was carried out in Deqing County, Zhejiang Province, from September 1981 to 1986 in order to determine whether such cases play a role in the continuation of transmission. The results of parasitological and entomological investigations for two consecutive years revealed that after the implementation of intervention measures, the mean microfilaraemia rate in the population fell to about 0.5% and the mean microfilaria density to about 4.2 mf/60 microliter of blood in previously endemic areas of malayan filariasis. Although there were considerable numbers of An. sinensis biting humans, infective larvae could be found in only two positive mosquitoes out of 5,484 dissected, and no new microfilaraemic cases were detected in the 1983 and 1986 follow-up blood examinations, indicating that transmission had already been interrupted. Two volunteers with a microfilaraemia of 3-5 mf/60 microliter of blood were exposed to two batches of An. sinesis in August 1981. The engorged mosquitoes were dissected eight days later. Even though the infection rate of An. sinensis having fed on low-density microfilaraemic cases was as high as 16.8%, the intensity of infection was extremely low, being 1.1 mf/mosquito. From the transmission dynamics point of view, infected mosquitoes carrying very few infective larvae have no practical significance in the transmission of filariasis. It is suggested that the treatment of persons with low-density microfilaraemia (with 5 mf/60 microliters of blood) in areas with low microfilaria rates (less than 1%) need not to be considered as essential.     

Identification of mosquito blood meals by enzyme-linked immunosorbent assay

A micro enzyme-linked immunosorbent assay for identifying mosquito blood meals is described. Antisera for the assay were prepared in rabbits and guinea pigs and purified by precipitation with cross-reacting blood sera. Positive blood source identifications could be made to the generic level using as little as 58 micrograms of ingested blood serum. The blood meals of 37 of 38 laboratory mosquitoes fed on seven host animals were correctly identified, including a mosquito that contained a double feed.     

The effect of diethylcarbamazine treatment on the development of bancroftian microfilariae in Culex p. fatigans.

Bancroftian microfilarie survived after one to three courses of diethylcarbamazine treatment in carriers and the larvae able to reach the infective stage in Culex p. fatigans. The infection rate and the development of infective larvae per infected mosquito from DEC-treated carriers was much lower than from the untreated carrier. The fact that surving MF can develop to infective stage in their vector indicates that such filarial carriers may be important sources for transmission of filariasis in the endemic areas after suspension of control measures.     

Mortality and infertility in adult mosquitoes after the ingestion of blood containing ivermectin

Mosquitoes of 3 species (Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus) were fed on human blood containing various concentrations of ivermectin. Three effects (death, decreased egg production, and reduced egg hatching) were observed in the insects, depending upon the concentration of ivermectin ingested. The LD50 of ivermectin in human blood for the 3 mosquito species was estimated to be 126, 208, and 698 ng/ml, respectively. Mosquitoes dying after ingestion of ivermectin developed signs of acute toxicity including paralysis, lethargy, incoordination, and difficulty in movement. Death usually occurred within 48-72 hr. With sublethal blood concentrations of the chemical, mosquitoes survived, but there was a marked reduction in both the number and viability of their eggs. This infertility was only temporary, however, as subsequent refeeding of the insects on uncontaminated blood resulted in the production of normal numbers of fertile eggs. Blood levels of ivermectin which made 50% of the eggs infertile in Ae. aegypti and Ae. albopictus were calculated 3.4 and 4.3 ng/ml, respectively. These latter concentrations of the chemical are within the range found in blood of humans and domestic animals receiving ivermectin for treatment of parasitic infections. This finding suggests that the widespread use of ivermectin in veterinary and human medicine may have an unrecognized effect on mosquito populations.     

Blood meal size as a factor affecting continued host-seeking by Aedes aegypti (L.)

The effect of ingested blood on the host-seeking response of two strains of Aedes aegypti was examined. Using an olfactometer, females fed partial blood meals were scored for host-seeking behavior within 1 h, and their blood meal sizes were measured chemically immediately afterwards. The suppression of host-seeking within 1 h after a blood meal appears to be caused by abdominal distention from ingested blood. Mosquitoes of either strain were attracted to a host when the blood meal size was less than 2.5 microliter; above this threshold there was a sharp decline in the tendency to respond. Small mosquitoes resulting from a low larval diet had a lower threshold, and were more likely to cease host-seeking after a small blood meal. Multiple feeding within a single gonotrophic cycle may result if mosquitoes take small blood meals which are insufficient to terminate host-seeking. Partial meals and reduced feeding success of mosquitoes can result from defensive host behavior, which in the laboratory rat was shown to increase at high mosquito densities.     

Effects of ivermectin and diethylcarbamazine on microfilariae and overall microfilaria production in bancroftian filariasis

Ivermectin and diethylcarbamazine (DEC) are used in mass treatment programs for the elimination of lymphatic filariasis because of their strong effects on microfilaremia. However, the effects of treatment on adult worms and the degree of individual variation in efficacy are unclear. We analyzed series of microfilaria (Mf) counts from individuals treated with a single dose of 400 microg/kg ivermectin or 6 mg/kg DEC (N = 23 in each group; 1 year follow-up). For each individual, we estimated the microfilaricidal effect and the reduction in overall Mf production (e.g., caused by death or sterilization of worms, or inhibited Mf release from the female worm uterus). Ivermectin on average killed 96% of Mf and reduced Mf production by 82%. DEC killed 57% of Mf and reduced Mf production by 67%, with some individuals responding very poorly. The strong reduction in overall Mf production is good news for control of lymphatic filariasis, but the prospects of elimination will be diminished if part of the population systematically responds poorly to treatment.     

A randomized clinical trial comparing single- and multi-dose combination therapy with diethylcarbamazine and albendazole for treatment of bancroftian filariasis

The Global Program for Elimination of Lymphatic Filariasis calls for mass drug administration for endemic populations outside of sub-Saharan Africa with a single dose of diethylcarbamazine (DEC) and albendazole (Alb) annually for 4-6 years. Single-dose DEC/Alb dramatically reduces blood microfilaria (MF) counts, but most treated subjects fail to completely clear MF after a single dose. A more effective regimen might reduce the number of years required for elimination programs. We performed a randomized clinical trial in Egyptian adults with asymptomatic microfilaremia to compare treatment with seven daily doses of oral DEC (6 mg/kg) and Alb (400 mg) with a single dose of the same combination. We also studied the effect of re-treatment with single-dose DEC/Alb 12 months after the first treatment course. Multi-dose DEC/Alb was significantly more effective than single-dose therapy for reducing and clearing microfilaremia (mean reduction in MF/ml relative to pretreatment counts at 12 months, 99.6% versus 85.7%, with complete clearance in 75% versus 23.1%). The two regimens had similar activity against adult filarial worms, as indicated by serial ultrasound assessments. Neither regimen resulted in complete clearance of filarial antigenemia. There was no difference in adverse events, which were mild to moderate. Blood microfilaria and parasite antigen clearance rates increased following re-treatment. Multi-dose DEC/Alb may be a useful option for filariasis elimination programs, especially in the first year (when enthusiasm for mass drug administration and coverage rates are high), to quickly reduce community MF loads and transmission rates.     

Mosquito infection studies with Aotus monkeys and humans infected with the Chesson strain of Plasmodiun vivax

Oocyst counts were compared between mosquitoes that fed on humans versus mosquitoes that fed on Aotus monkeys, both of which were infected with the Chesson strain of Plasmodium vivax. Oocyst counts obtained from mosquitoes fed on humans were almost 10-fold higher in number. Mosquitoes were more likely to be infected and with a higher rate of infection when they fed on monkeys before the peak in the asexual parasite count. Mosquitoes that fed on humans were more likely to be more heavily infected when fed after the peak in the asexual count. Of several species of owl monkeys, Aotus vociferans was infected at a higher frequency. On the basis of oocyst counts, Anopheles dirus were the most susceptible and An. maculatus were the least susceptible of the mosquito species tested.     

河南省消除丝虫病的监测与审评

目的消除丝虫病残存传染源。方法1995~2002年应用厚血膜法和蚊媒解剖法,对全省69个原丝虫病流行县市实施纵、横向病原学和蚊媒监测。2003年依照卫生部消除丝虫病审评办法,对全省各丝虫病流行县进行了消除丝虫病达标审评。结果全省69个县市实施消除丝虫病监测,总监测乡镇数覆盖原丝虫病流行乡镇38.58%(以县为单位30%~100%),总监测人口覆盖流行区人口3.92%(以县为单位在3%~21.76%之间)。全省390个蚊媒监测点共捕获、剖检媒介蚊虫245878只,其中淡色库蚊213746只、中华按蚊32132只;各流行县市均达到了部颁消除丝虫病蚊媒监测标准。未发现微丝蚴血症者和幼丝虫阳性蚊。结论通过对全省69各流行县市的丝虫病达标监测和审评,证实河南省已经消除丝虫病传染源,达到了卫生部颁消除丝虫病标准。     

Indoor resting density, survival rate and host preference of Culex quinquefasciatus say (Diptera: Culicidae) in Mysore City.

Monthly indoor resting density, age distribution, survival rate and host-preference of Culex quinquefasciatus, a filariasis vector were studied from June 1988 to May 1989 in Mysore city. During this period, a total of 7522 adults were collected in 72 man-hour of search spreading to twelve spots covering the entire city. The annual per man-hour density observed was 104.5, being 76.8 for female and 24.3 for male. Nulliparous and parous (1-parous and 2-parous) mosquitoes were collected throughout the year while 3-parous and 4-parous mosquitoes were recorded during cold and rainy seasons only showing an increase in the number of gonotrophic cycles. Precipitin test of blood smears from the gut content of C. quinquefasciatus was done by taking 2536 mosquito blood smears. Of these, 94.1 per cent preferred human blood, 1.4 per cent bovine and 3 per cent avain. As the species evaluated is a major vector of filariasis, the present study on its resting density and the host preference will be helpful in vector monitoring in Mysore city. Various results on this species at Mysore have been discussed in the light of earlier studies conducted elsewhere.     

Human Host-Derived Cytokines Associated with Plasmodium vivax Transmission from Acute Malaria Patients to Anopheles darlingi Mosquitoes in the Peruvian Amazon

Infection of mosquitoes by humans is not always successful in the setting of patent gametocytemia. This study tested the hypothesis that pro- or anti-inflammatory cytokines are associated with transmission of Plasmodium vivax to Anopheles darlingi mosquitoes in experimental infection. Blood from adults with acute, non-severe P. vivax malaria was fed to laboratory-reared F1 An. darlingi mosquitoes. A panel of cytokines at the time of mosquito infection was assessed in patient sera and levels compared among subjects who did and did not infect mosquitoes. Overall, blood from 43 of 99 (43%) subjects led to mosquito infection as shown by oocyst counts. Levels of IL-10, IL-6, TNF-α, and IFN-γ were significantly elevated in vivax infection and normalized 3 weeks later. The anti-inflammatory cytokine IL-10 was significantly higher in nontransmitters compared with top transmitters but was not in TNF-α and IFN-γ. The IL-10 elevation during acute malaria was associated with P. vivax transmission blocking.     

The impact of six rounds of single-dose mass administration of diethylcarbamazine or ivermectin on the transmission of Wuchereria bancrofti by Culex quinquefasciatus and its implications for lymphatic filariasis elimination programmes

Lymphatic filariasis (LF) is targeted for global elimination. Transmission interruption through repeated annual single-dose mass administration of anti-filarial drugs is the mainstay of the LF elimination strategy. This study examined the ability of six rounds of mass administration of diethylcarbamazine (DEC) or ivermectin (IVM) to interrupt transmission of Wuchereria bancrofti by Culex quinquefasciatus, the predominant parasite and vector species, respectively. After six rounds of mass drug administration (MDA), received by 54-75% of the eligible population (> or =15 kg body weight), the resting vector infection and infectivity rates fell by 83% and 79% in the DEC arm, 85% and 84% in the IVM arm and 31% and 45% in the placebo arm, respectively. The landing vector infection and infectivity rates fell by 83% and 94% in the DEC arm, 63% and 75% in the IVM arm and 1% each in the placebo arm, respectively. The filarial larval load per resting mosquito declined by 92% and 93% and per landing mosquito by 83% and 69% in the DEC and IVM arms, respectively. The annual infective biting rate (AIBR) fell from 735 to 93 (87%) in the DEC arm, 422 to 102 (76%) in the IVM arm and 472 to 398 (16%) in the placebo arm. The annual transmission potential (ATP) declined from 2514 to 125 (95%), 1212 to 241 (80%) and 1547 to 1402 (9%) in the DEC, IVM and placebo arms, respectively. However, mosquitoes with infection [microfilaria/larva 1/larva 2 (Mf/L1/L2)] were found in all study villages. Three of five villages in the IVM arm and two of five in the DEC arm recorded no resting mosquitoes with infective-stage (L3) larva. Although the ATP, after six rounds of MDA, fell substantially and remained at 125 and 241 in the DEC and IVM arms, respectively, the cumulative exposure to infective stage larvae (ATP) during the treatment period of 6 years was as high as 2995 in the DEC arm and 1522 in the IVM arm, because of considerable level of transmission during the initial (1-3) rounds of MDA. We conclude that (i) six rounds of MDA, even with 54-75% treatment coverage, can reduce LF transmission very appreciably; (ii) better treatment coverage and a few more rounds of MDA may achieve total interruption of transmission; (iii) high vector densities may partly nullify the reductions achieved in vector infection and infectivity rates by MDA and (iv) achievement of 'true zero' Mf prevalence in communities and 0% infection rate (mosquitoes with Mf/L1/L2) in mosquitoes may be necessary to totally interrupt Culex-transmitted LF.     

Persistence of Brugia malayi DNA in vector and non-vector mosquitoes: implications for xenomonitoring and transmission monitoring of lymphatic filariasis

Xenomonitoring (detection of filarial larvae or their DNA in mosquitoes) is a sensitive marker for assessing the endemicity of filariasis and a useful tool for evaluating elimination programs. To examine the fate of microfilariae (mf) and filarial DNA in vector competent and non-competent mosquito strains, we compared the detection of Brugia malayi parasites by dissection and by quantitative real-time polymerase chain reaction (PCR) in three different mosquito strains. We conclude that PCR is much more sensitive than dissection for detecting filarial larvae, especially their remnants in mosquitoes. However, parasite DNA can be detected in both vector and non-vector mosquitoes for two weeks or longer after they ingest mf-positive blood. Thus, although xenomonitoring with vector and non-vector mosquito species may be a sensitive method for indirectly detecting filarial parasites in human populations, positive test results for parasite DNA in mosquitoes do not necessarily prove that transmission is ongoing in the study area.