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PubMed2006

Chronic clinical manifestations related to Wuchereria bancrofti infection in a highly endemic area in Kenya.

Clinical examinations were conducted in an effort to provide baseline data for a pilot filariasis elimination programme implemented in a Wuchereria bancrofti-endemic focus in Malindi district, Kenya. Of 186 males aged 15 years and above examined, 64 individuals (34.4%) had hydrocele, and the prevalence of the manifestation in those above 40 years old was 55.3%. The prevalence of leg lymphoedema in persons aged 15 years and above was 8.5%, with a higher rate in males (12.6%) than in females (5.7%). The overall prevalence of inguinal adenopathy was 8.6%, and males had a significantly higher (12.9%) prevalence of adenopathy than females (5.1%) (P<0.001). The data in the present study provided support for consideration of filarial infection as a possible cause of inguinal lymphadenopathy in bancroftian filariasis-endemic areas. The results of this study also indicate that lymphatic filariasis is a serious public health problem in the northern coastal areas and morbidity control programmes should be implemented to alleviate the suffering of those affected.

Adolescent↗
PubMed2002

Genetic variability of the human filarial parasite, Wuchereria bancrofti in South India.

The genetic variability of the lymphatic filarial parasite Wuchereria bancrofti, from three localities (one urban and two rural areas) in southern India, endemic for filariasis was studied using random amplified polymorphic DNA (RAPD) markers. The RAPD profiles were generated for 21 parasite populations (7 populations from each area), using a 10-mer random primer. The analysis of profiles indicated the existence of considerable genetic variability among parasite populations. The Nei's gene diversity between the individual populations in the 2 areas (one urban and another rural) was comparatively greater (0.3372+/-0.1462 & 0.2830+/-0.1764) than that of populations in another village (0.0490+/-0.1373). The greater genetic diversity among the former areas may be due to human migration, endemicity for long time and drug (diethyl-carbamazine citrate) pressure unlike the populations of latter village where the filariasis is relatively a recent introduction and which was never under active chemotherapy. The Nei's genetic distance was estimated and the phylogenetic tree was constructed using 'UPGMA'. These analyses indicated the prevalence of at least two genetically distinct clusters, among the populations studied, their maximum genetic distance being 0.2444. The finding of two genetic 'variants' of W. bancrofti, in the present study, may have important implications in filariasis epidemiology and control/elimination programmes.

Animals↗
PubMed2003

A simple and rapid DNA extraction method for the detection of Wuchereria bancrofti infection in the vector mosquito, Culex quinquefasciatus by Ssp I PCR assay.

A simple, rapid and inexpensive method for the extraction of DNA from filarial vector, Culex quinquefasciatus, useful in Ssp I PCR assay for xenomonitoring of infection with Wuchereria bancrofti is presented. The DNA extracted by this method was found suitable for PCR detection of W. bancrofti infection in pools of 10-30 mosquitoes. The PCR assay employing the simplified DNA extraction method was evaluated for its sensitivity on field caught Cx. quinquefasciatus, in comparison with the conventional dissection and microscopy technique. When assayed on dissection washings of vector mosquitoes the PCR assay detected 45 pools out of 49 dissection positive pools as positive for infection and hence found to be less sensitive than the conventional technique. The reason for detecting four dissection positive pools as negatives by the PCR assay may be due to the loss of a few numbers of parasites (1-3) present in these pools during the transfer of washings of dissected mosquitoes. The PCR assay detected ten out of 72 dissection negative pools as positives, while it did not detect any of the 62 known negative (laboratory reared, uninfected) mosquito pools as positives. When 38 pools (10 mosquitoes/pool) of intact mosquitoes were assessed for infection by each method, the infection rates obtained by the two methods were almost similar (3.35 and 3.01%, respectively, for conventional method and PCR assay). The results thus show that the DNA extraction method, which is simple, rapid, safe and inexpensive, is efficient to generate DNA from vector mosquitoes useful in PCR assay and hence has potential application in xenomonitoring.

Animals↗
PubMed2003

Detection of codon for amino acid 200 in isotype 1 beta-tubulin gene of Wuchereria bancrofti isolates, implicated in resistance to benzimidazoles in other nematodes.

Albendazole, anthelmintic benzimidazole (BZ) is being co-administered with an antifilarial drug, diethylcarbamazine (DEC), in lymphatic filariasis (LF) elimination programmes. But this drug is known to result in the faster development of drug resistance and hence it is necessary to monitor drug sensitivity among populations of Wuchereria bancrofti. In vitro toxicity assays to detect drug sensitivity are not available for this parasite and hence alternative techniques such as PCR assays need to be developed, for which information on genetic basis for drug sensitivity/resistance is necessary. Such information is lacking in the case of W. bancrofti, but available for nematode parasites of farm animals wherein it has been established that replacement of phenylalanine with tyrosine at amino acid position 200 of beta-tubulin isotype 1 conferred BZ-resistance. Using this information we examined the polymorphism in the codon of this residue in W. bancrofti populations representing geographically distant areas of India, through sequencing exon 5 region of beta-tubulin isotype 1 gene. The nucleotide sequence data showed that W. bancrofti isolates from wide geographic areas of India had codon for Phe (TTC) at position 200 of the beta-tubulin isotype 1 gene suggesting that the parasite might be genetically sensitive to BZ.

Albendazole↗
PubMed2000

Development of Wuchereria bancrofti in Culex quinquefasciatus that survived the exposure of sub-lethal dose of Bacillus sphaericus as larvae.

Development of Wuchereria bancrofti in Culex quinquefasciatus emerged from the larvae that survived the exposure of sub-lethal dose of Bacillus sphaericus was examined in the laboratory. Third instar larvae of Cx. quinquefasciatus were treated with B. sphaericus at a sub-lethal dose of 11.35 microg/250 ml. The female mosquitoes that emerged from the survived larvae were fed on microfilaraemic human blood and parasite development was monitored in the fed mosquitoes. Both treated and untreated mosquitoes could ingest microfilaria (mF) equally as there was no significant difference in mF density between them. But, density of developmental stages of the parasite in treated group was significantly lower. Since, there was no mortality of mosquitoes, the lower density of the developmental stages could be attributed to the loss of parasites in the treated mosquitoes. Consequently, the proportion of mosquitoes with infective larvae (L3) and number of L3 were also significantly lower in treated females. Delay in parasite development was also noticed in treated mosquitoes. The present study indicates that B. sphaericus, when applied at sub-lethal dose kills larvae, and in addition, inhibits development of the filarial parasite and consequently reduces L3 yield in adult mosquitoes that emerged from the survived larvae.

Animals↗
PubMed2002

Red blood cell antioxidant levels in Wuchereria bancrofti infection.

The elimination of microfilariae of Wuchereria bancrofti is probably mediated by free radicals. Red cell catalase (C), glutathione peroxidase (GPX), and superoxide dismutase (SOD) activity levels were measured as an indirect method of assessing blood oxidant status in 29 asymptomatic microfilaraemics, 29 "endemic normals", and 29 controls living in a non-endemic area. Changes in the activity of these enzymes were also compared over a one month period in 22 asymptomatic microfilaraemics randomised to receive either single dose or 14 day treatment with diethyl carbamazine citrate (DEC). Red cell GPX activity levels were significantly higher in "endemic normals" when compared to mf positive cases and non-endemic controls. An early and significant increase in GPX activity (on days 3, 7 and 14 compared to pretreatment levels, p<0.01) was observed after DEC in both treatment groups. Increases in the activity of catalase and SOD became significant only on days 14 and 30 respectively. The percentage reduction in microfilaraemia correlated significantly with the percentage increase in GPX activity levels (R(2)=0.58, p=0.6 x 10(-5)). Our results may suggest a role for GPX related oxidant species in the elimination of microfilariae.

Adolescent↗
PubMed2002

Age-grading and growth of Wuchereria bancrofti (Filariidea: Onchocercidae) larvae by growth measurements and its use for estimating blood-meal intervals of its Polynesian vector Aedes polynesiensis (Diptera: Culicidae).

Growth in length and width of Wuchereria bancrofti (Filariidea: Onchocercidae) larvae developing in its Polynesian vector Aedes polynesiensis (Diptera: Culicidae) was analysed using a mathematical approach to objectively extract patterns. L1 had a U-shaped growth in length, while widths followed an S-shaped function. L2 had an S-shaped growth in length and width. Growth in length of L3 was also S-shaped, while widths had an asymptotic size following a period of rapid shrinkage. The greatest difference between length and width was in stage 3 where the length was over 75 times greater than the width. The ratio of length to width was approximately 50 for microfilariae and only 10 for the L1 ('sausage') stage. Characteristic mean length (and width) were approximately 280(7) microm for microfilariae, approximately 181 microm for L1 at their smallest, and approximately 1584(22) microm for L3 infective larvae. There was a great increase in length during stage 2 from approximately 322(27) to approximately 982(31) microm. Stage duration decreased with increasing temperature while growth rate increased, giving steeper growth curves. There was no effect of temperature on size, except for L3, which were shorter when mosquitoes were reared at higher temperature. It appears that larval growth is a continuous process from microfilariae to the young L3 stage, and continuously modifies the larval parasite aspect, even within each stage. Thus, information on larval shape may be used as an age indicator and in some cases, may give an estimation on time elapsed since infection of the vector. An important demographic parameter used in most mathematical models describing transmission of parasites by insect vectors is the length of the gonotrophic cycle of the vector, i.e. the time interval between two successive blood-meals. Usual methods for computing such a parameter are based on mark-recapture techniques. However, reliable estimates need substantial capture rates, which are not always possible. This paper presents another approach in which marked mosquitoes are those naturally infected by W. bancrofti. For one mosquito, the time since infection is simply the age of the developing larval parasite. Our method first expresses the age of larval parasite as a fraction of total development time (from microfilariae entering the vector to L3 larvae) using a regression model based on measurements of the parasite's length and width. This fraction of development is then converted to a chronological age since infection, using a back-calculation procedure involving ambient temperatures and growth rates of W. bancrofti larvae in the vector. The method is applied to wild caught Ae. polynesiensis in French Polynesia to compute the length of the gonotrophic cycle. This mosquito species comes to bite approximately 3, 6-7 and 9 days after a first infectious blood-meal. Then the length of the gonotrophic cycle may be of 3-4 days.

Aedes↗
PubMed2000

Molecular characterisation of the actin gene of the filarial parasite Wuchereria bancrofti.

Wuchereria bancrofti is the major cause of lymphatic filariasis in humans. Although it is responsible for this immensely morbid and debilitating disease, very little is known of the basic molecular biology of this parasite, and there is a vast lack of knowledge on its gene organisation. In this study, the actin gene of W. bancrofti has been characterised by sequencing a clone isolated from a genomic DNA library of this parasite. The 5' flanking region had a potential TATA box and a putative mRNA initiation site. The gene had five exons encoding 376 amino acids, and four introns ranging in size from 109 to 190bp. The 3' flanking region had a potential polyadenylation signal with the sequence ATTAAA which is a common natural variant of the conventional sequence AATAAA. The gene was AT-rich, with a GC content of 37.2%. Southern blot analysis of W. bancrofti genomic DNA indicated that the gene is possibly found as a single copy. The actin amino acid sequence of W. bancrofti showed a high degree of homology to the actin of many organisms of different taxonomic groups, but the highest homology was observed with the free-living nematode Plectus acuminatus. This suggests that P. acuminatus may bear a close evolutionary relationship to W. bancrofti.

Actins↗
PubMedDate not supplied

Genital manifestations and reproductive health in female residents of a Wuchereria bancrofti-endemic area in Tanzania.

To assess the significance of lymphatic filariasis for the development of chronic genital manifestations and for reproductive health in women, we conducted a cross-sectional study of 2 villages in north-eastern Tanzania including interview and gynaecological examination of adult women, focusing primarily on reproductive history and genital health. In a population of 2165 residents, prevalence of Wuchereria bancrofti microfilaraemia was 28%, and geometric mean intensity of microfilariae (mf) was 722 mf/mL. Leg lymphoedema (elephantiasis) was present in 4.2% of adults aged > or = 15 years, and hydrocoele in 26.5% of adult males. Five hundred and thirty women completed an interview, and 404 of these completed a gynaecological examination. Most women were Muslims (72%), polygamy was common (29%), and 49% of women had undergone circumcision (clitoridectomy). Presence of microfilaraemia did not influence fertility or fertility-related variables including age at menarche, parity, spontaneous abortion, stillbirth, Caesarean section and premature labour, nor presence of primary or secondary infertility. Mf status was strongly correlated with abnormal menstruation pattern in the 30+ years age-group (P = 0.001), but not in the < 30 years age-group. Cervical, vaginal and vulval pathology was unrelated to mf status. Two women, aged 46 and 77 years, had vulval oedema of probable filarial origin, both were mf negative. Overall, microfilaraemia appeared to have no influence on genital disease or reproductive health, and chronic manifestations of lymphatic filariasis of the genitals does not appear to be a substantial problem in women.

Adolescent↗
PubMedDate not supplied

Single-dose treatment of Wuchereria bancrofti infections with ivermectin and albendazole alone or in combination: evaluation of the potential for control at 12 months after treatment.

The effect of single-dose ivermectin (150-200 micrograms/kg) and albendazole (400 mg) treatment alone and in combination on Wuchereria bancrofti microfilaraemia, antigenaemia and clinical manifestations was compared 12 months after treatment in a double-blind placebo-controlled field trial carried out in Ghana in 1996-98, to evaluate the potential of these treatments for control. Both ivermectin and combination treatments resulted in pronounced reductions in microfilaraemia among individuals who were microfilaria positive before treatment. Among individuals who were positive for circulating filarial antigen before treatment, antigen levels increased considerably over the 1-year period after treatment in the placebo group, whereas they decreased in the ivermectin and combination groups. However, the post-treatment difference reached statistical significance in neither microfilaraemia nor antigenaemia between the ivermectin and the combination groups. Albendazole treatment alone showed only a minor effect on microfilaraemia and antigenaemia. No effect of the treatments on the incidence of new cases of microfilaraemia or antigenaemia, or on clinical manifestations, was observed. Both ivermectin and combination treatment thus appeared effective for control of W. bancrofti infections, but the difference in efficacy between the 2 treatments after 12 months appeared to be minimal.

Adolescent↗
PubMedDate not supplied

The transmission dynamics of bancroftian filariasis: the distribution of the infective larvae of Wuchereria bancrofti in Culex quinquefasciatus and Anopheles gambiae and its effect on parasite escape from the vector.

The anatomical distribution of the infective larvae (L3) of Wuchereria bancrofti in Culex quinquefasciatus and Anopheles gambiae, and its influence on L3 escape, was evaluated by exposing the vectors to human individuals infected with W. bancrofti. After the extrinsic incubation period of W. bancrofti, a random sample of the infected mosquitoes was dissected to determine the distribution of infective larvae in the body of the mosquitoes and the proportion of mosquitoes that were infected. The remaining mosquitoes were exposed to an experimental definitive host (mouse skin). The infective larvae on and in the host tissues were counted. The engorged mosquitoes were dissected to determine the proportion infected and the distribution of L3 in mosquitoes after exposure. The results show that 53.4% of L3 escaped from Cx. quinquefasciatus to enter the experimental host whereas only 4% of L3 escaped from An. gambiae. Analysis of the results indicates that the number of L3 in the vector and the proportion of L3 in the head and mouthparts influence the number of L3 escaping from the vector to enter the definitive host and consequently the number that gain access to host tissues. The implication of these findings on the transference of the parasite from the vector to the definitive host is discussed.

Animals↗
PubMedDate not supplied

Ultrastructural characterization of intracellular bacteria of Wuchereria bancrofti.

Ultrastructural observations on the structure and distribution of endosymbiotic bacteria within the tissues of Wuchereria bancrofti are described. In female worms the organisms were observed in the lateral cords of the hypodermis, oocytes, developing eggs and in intrauterine microfilariae. Organisms were also detected in blood microfilariae and in the intestine of third-stage larvae. Bacteria were not observed in male worms.

Animals↗
PubMedDate not supplied

Detection of Wuchereria bancrofti in mosquitoes by the polymerase chain reaction: a potentially useful tool for large-scale control programmes.

Focally endemic bancroftian filariasis is targeted for elimination in the Nile delta of Egypt. Improved methods are needed for identifying endemic villages to be included in the control programme and for monitoring its success. We have evaluated the performance of a polymerase chain reaction (PCR) assay in estimating Wuchereria bancrofti infection in pools of Culex pipiens (1-25 females) from 2 adjacent villages with high (El Qolzom, 10.8%) and low (Kafr Shorafa, 2.1%) prevalence rates of human filariasis. This assay detects a repeated sequence in W. bancrofti deoxyribonucleic acid (DNA). Mosquitoes resting within houses were captured by aspiration and pooled by house. Houses were classified as positive or negative for human filarial infection based on night blood examinations of residents. The assay detected parasite DNA in mosquitoes from 60% of 25 infected houses and 24% of 25 uninfected houses. PCR processing of mosquitoes caught within houses of unknown filariasis infection status (44 in El Qolzom, 37 in Kafr Shorafa) identified 31.8% and 8.1% of houses, respectively, as containing infected mosquitoes. These results support the validity of the PCR assay for evaluating filarial prevalence in different villages. C. pipiens collected outdoors in dry ice-baited traps and tested by PCR (266 in Qolzom, 82 in Kafr Shorafa) did not contain parasite DNA. Pools of female mosquitoes (296 in Qolzom, 240 in Kafr Shorafa) captured in oviposition traps were also negative. We concluded that the PCR based assay is a powerful epidemiological tool that can be used for evaluating W. bancrofti infection in villages in the Nile delta and for monitoring the application of control programmes in filariasis endemic areas.

Animals↗
PubMedDate not supplied

Prevalence and intensity of Wuchereria bancrofti antigenaemia in Sri Lanka by Og4C3 ELISA using filter paper-absorbed whole blood.

In Sri Lanka 2741 people from Matara, an endemic area for Wuchereria bancrofti, were examined in 1996/97 for microfilariae by 60-microL blood smear and for circulating filarial antigens by Og4C3 ELISA using filter paper-absorbed whole blood. The overall prevalence of microfilaraemia was 3.4%, and that of antigenaemia 14.4%. The prevalence of antigen-positive and microfilaria-negative people was 11.3%. Analysed by age-group, antigenaemia prevalence was similar in all groups, and the average number of antigen units was already very high in the age-group < 10 years, indicating that the infection started in early childhood. Among those who were antigen positive, the microfilaria prevalence was lower in females than in males. Diethylcarbamazine treatment eliminated microfilariae in 78% of the positives. However, 17 months after the treatment, antigenaemia was still positive in 76% of those who were parasitologically cured.

Adolescent↗
PubMedDate not supplied

Progression of lymphatic vessel dilatation in the presence of living adult Wuchereria bancrofti.

Bancroftian filariasis, a mosquito-transmitted disease commonly known as elephantiasis, is caused by infection with the parasite Wuchereria bancrofti. Infection with this parasite can induce a broad array of chronic debilitating and socially stigmatizing conditions, but the pathogenesis of this morbidity remains obscure. Recent evidence indicates that in filariasis-endemic areas the primary lesion is not lymphatic vessel obstruction but, rather, dilatation. To determine the extent to which lymphatic dilatation occurs in the presence of living adult W. bancrofti, we performed longitudinal ultrasonographic measurements in 80 men (mean age 24 years) in Brazil who had a total of 107 W. bancrofti nests detectable by ultrasound. Initial mean lymphatic vessel diameter at the site of the worms was 3.4 mm (range, 0.7-11.3), and was greater in men with 2 or more nests (3.9 mm) than in those with only one nest (3.0 mm, P = 0.003). During the study period (2-35 months, mean, 13.7), lymphatic vessel diameter increased at the site of 92 (86.0%) adult worm nests. Mean rate of increase of lymphatic vessel diameter was 1.2 mm per person-year (range, 0-0.93 mm per month). In a general linear model, no factors, including treatment with antifilarial drugs, were significantly associated with rate of vessel diameter increase. Thus, lymphatic vessel dilatation progress in the presence of living adult W. bancrofti; the rate of this progression is heterogeneous. These data suggest that lymphatic dilatation will continue to progress in most infected persons even after mass treatment with currently recommended antifilarial drugs. In addition to interrupting transmission, the global programme for elimination of lymphatic filariasis should address the potential for disease progression in persons who remain infected with adult W. bancrofti.

Adolescent↗
PubMedDate not supplied

Pathogenesis of filarial hydrocele: risk associated with intrascrotal nodules caused by death of adult Wuchereria bancrofti.

Although testicular hydrocele is the most common clinical manifestation of bancroftian filariasis, its pathogenesis is poorly understood, as is its relationship to inflammatory scrotal nodules following death of adult Wuchereria bancrofti. Between 1994 and 1998, we prospectively determined the incidence and clinical evolution of nodule-associated acute hydrocele in men attending 2 outpatient clinics in Recife, Brazil who were infected with W. bancrofti, had living adult worms detectable by ultrasound in the intrascrotal lymphatic vessels, and were scheduled for treatment with 6 mg/kg diethylcarbamazine (DEC). A total of 132 men developed 173 scrotal nodules 1-7 (mean 4.2) d after DEC treatment and another 47 developed 58 spontaneous nodules before they received DEC treatment. These 179 men with a single 'nodule event' (simultaneous development of > or =1 scrotal nodules) were followed-up by serial physical and ultrasound examinations for 18 months. Overall, 40 (22.3%) men developed acute hydrocele, 3 of whom underwent biopsy and hydrocele repair. Of the remaining 37 men, 9 (24.3%) developed chronic hydrocele and 28 had acute hydrocele resolution within 14-210 (mean 60.9) d. Rate of chronic hydrocele was similar for men who received DEC and those with spontaneous nodules. Seventeen (42.5%) men with hydrocele had multiple scrotal nodules, compared with 28 (20.1%) men who did not develop hydrocele (P= 0.007). Of 134 men with single nodules, superior paratesticular nodules were found in 56.5% and 29.7% of those with and without hydrocele, respectively (P = 0.02). Acute hydrocele occurs frequently following death of adult W. bancrofti and single episodes of scrotal nodule formation. Chronic hydrocele may develop following 5.1% of these episodes.

Acute Disease↗
PubMedDate not supplied

Age-dependent prevalence of asymptomatic amicrofilaraemic individuals in a Wuchereria bancrofti-endemic region of India.

The age-related prevalence of asymptomatic amicrofilaraemics, apparently uninfected individuals, was determined in a Wuchereria bancrofti-endemic region of Orissa, India during 2001. The prevalence of these uninfected individuals was highest in the younger age groups (aged < or = 15 years), decreased rapidly in those aged > 15 to 40 years, and then stabilized in those aged > 40 years.

Adolescent↗