Bancrofti Infection Research Articles

Wuchereria bancrofti in a community with seasonal transmission: stability of microfilaraemia, antigenaemia and filarial-specific antibody concentrations.

The effect of seasonal transmission on microfilaraemia, antigenaemia and filarial-specific antibody levels in individuals infected with Wuchereria bancrofti was investigated in a follow-up study in an endemic community in north-eastern Tanzania. The subjects were 37 adult male residents who were found to be positive for circulating filarial antigen (CFA) at the beginning of the study (26 of whom were also found microfilaraemic with W. bancrofti at this time). Blood samples were collected from each subject in July 1998, January 1999 and July 1999, during the seasons when transmission intensity was high, low and high, respectively. The mean intensities of microfilaraemia and the mean concentrations of CFA were each slightly higher during the low-transmission season than during the two high-transmission seasons but the differences were not statistically significant (P > 0.05). Similarly, the mean levels of filarial-specific IgG1, IgG2, IgG3, IgG4 or IgE did not differ to a statistically significant degree between the three examination times. Microfilaraemias and the levels of CFA and filarial-specific antibodies all therefore appeared to be remarkably stable and largely unaffected by the seasonal variation in transmission. That no variation in the mean IgG4/IgE ratio was observed over the study period may indicate that the level of resistance to W. bancrofti infection in the study subjects was also unaffected by the transmission season.

Wuchereria bancroftiin a community with seasonal transmission: stability of microfilaraemia, antigenaemia and filarial-specific antibody concentrations

The effect of seasonal transmission on microfilaraemia, antigenaemia and filarial-specific antibody levels in individuals infected with Wuchereria bancrofti was investigated in a follow-up study in an endemic community in north-eastern Tanzania. The subjects were 37 adult male residents who were found to be positive for circulating filarial antigen (CFA) at the beginning of the study (26 of whom were also found microfilaraemic with W. bancrofti at this time). Blood samples were collected from each subject in July 1998, January 1999 and July 1999, during the seasons when transmission intensity was high, low and high, respectively. The mean intensities of microfilaraemia and the mean concentrations of CFA were each slightly higher during the low-transmission season than during the two high-transmission seasons but the differences were not statistically significant (P > 0.05). Similarly, the mean levels of filarial-specific IgG1, IgG2, IgG3, IgG4 or IgE did not differ to a statistically significant degree between the three examination times. Microfilaraemias and the levels of CFA and filarial-specific antibodies all therefore appeared to be remarkably stable and largely unaffected by the seasonal variation in transmission. That no variation in the mean IgG4/IgE ratio was observed over the study period may indicate that the level of resistance to W. bancrofti infection in the study subjects was also unaffected by the transmission season.

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.

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 μg/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 1996–1998, 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.

The Wuchereria bancrofti orthologue of Brugia malayi SXP1 and the diagnosis of bancroftian filariasis

The gene encoding the Wuchereria bancrofti orthologue of the Brugia malayi-derived diagnostic antigen SXP1 was identified from a W. bancrofti L3 cDNA library and characterized. The Wb-sxp-1 cDNA encoded a basic protein with a calculated molecular mass of 20.8 kDa. Wb-SXP-1 was 85% identical to the SXP1 protein described from B. malayi ( Bm-SXP-1). The Wb-SXP-1 sequence also showed significant identity with proteins described from B. pahangi, Onchocerca volvulus, Acanthochilonema vitea, Ascaris suum, Loa loa, Litomosoides sigmodontis and Caenorhabditis elegans. The presence of a number of invariant and conserved residues in all of these nematode-derived molecules suggests that Wb-SXP-1 is a member of a new protein family. A recombinant form of Wb-SXP-1 was produced and it was determined that the anti- Wb-SXP-1 antibody response in patients with W. bancrofti infections was restricted to the IgG4 subclass. An anti- Wb-SXP-1 IgG4 ELISA was developed and this assay was found to be 100% sensitive for patients with patent W. bancrofti infection. Sera from individuals experiencing chronic pathology, endemic normals or patients with non-filarial nematode infections had no detectable IgG4 against Wb-SXP-1. While patients with patent Onchocerca volvulus infections were uniformly negative in the Wb-SXP-1 assay, 40% of sera from patent Loa loa infections were positive. When Bm-SXP-1 was used as the antigen under identical conditions, the assay was 88% specific for patent W. bancrofti infections and the antigen was recognized by antibodies from both O. volvulus and L. loa infections. The results strongly suggested that, for certain diagnostic filarial antigens, the use of same-species molecules can enhance the specificity of diagnostic tests.

Application of a polymerase chain reaction-ELISA to detect Wuchereria bancrofti in pools of wild-caught Anopheles punctulatus in a filariasis control area in Papua New Guinea.

Chemotherapy-based eradication programs are aimed at stopping transmission of Wuchereria bancrofti by its obligatory mosquito vector. This study compares one year post-treatment W. bancrofti infection rates of Anopheles punctulatus, the main vector of lymphatic filariasis in Papua New Guinea, using traditional dissection techniques and a polymerase chain reaction (PCR)-based ELISA of a parasite-specific Ssp I repeat. A total of 633 mosquitoes in 35 batches were dissected. Six batches contained W. bancrofti-infected mosquitoes, giving a minimum infection rate of 0.9%. This value was not different than the actual infection rate, which was 9 (1.4%) of 633 mosquitoes (P = 0.48). The DNA was extracted from 47 pools containing a mean of 13.2 mosquitoes per pool. A total of 621 mosquitoes were processed for the PCR-ELISA, including 486 caught by human bait and 135 by light trap, which included both dead and live mosquitoes. Of 23 pools of alcohol-preserved human-bait mosquitoes, seven were positive by the PCR-ELISA, giving an infection rate identical to that obtained by dissection of individual mosquitoes (1.4%). The minimum infection rates for pools of light-trap mosquitoes found dead and alive were 2.7% (2 of 74) and 4.9% (3 of 61), respectively. These values did not differ from each other (P = 0.84), but the overall infection rate of light-trap mosquitoes was greater than that of mosquitoes captured by human bait (3.7% versus 1.4%; P = 0.09). These data indicate that the PCR-ELISA of a W. bancrofti Ssp I repeat using pools of mosquitoes is comparable to traditional dissection techniques for monitoring transmission intensity following introduction of mass chemotherapy. This approach may also be useful for rapid and cost-effective assessment of transmission in endemic areas where the frequency of overt lymphatic pathology is low.

Apparent failure of ultrasonography to detect adult worms of Brugia malayi.

Adult worms of Wuchereria bancrofti, or rather their characteristic movements (the 'filarial dance'), can now be detected in the scrotal lymphatics of microfilaraemic males, using ultrasonography. This ability has been used to delineate the lymphatic pathology of bancroftian filariasis, guide the surgical removal of the adult worms and, most importantly, assess the macrofilaricidal effects of antifilarial drugs. In the present study, the first report of the use of ultrasonography in brugian filariasis, 22 men (aged 18-62 years) with 60-2972 (median = 370) Brugia malayi microfilariae/ml blood were subjected to ultrasonography using a linear, 7.5-MHz probe. In addition, four other men (aged 19-35 years), with W. bancrofti microfilaraemia [28-524 (median = 234) microfilariae/ml], were similarly examined. Adult worms were not detectable in any of the patients with B. malayi parasitaemia but were detected in the scrotal lymphatics of two of the four individuals with W. bancrofti infection. The reasons for the failure to detect adult B. malayi and the limitations of ultrasound as a screening tool are examined. The results highlight the differences between the two species that cause most lymphatic filariasis and the need for rapid development of tools that can be used for the control of brugian lymphatic filariasis.

Molecular and immunodiagnosis of human filarial nematode infections.

The filarial nematodes Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus represent major public health problems in the Tropics. Effective diagnosis of infection with these parasites is required both for administration of drugs to infected individuals and for monitoring of control programs. However parasitological diagnosis is associated with a number of problems including frequently inadequate sensitivity, long pre-patency of infection and inconvenience for patients. For these reasons there has been considerable effort expended in developing other forms of diagnosis, in particular immunoassays for measuring antibody and circulating parasite antigen as well as molecular-biology-based assays for detecting parasite DNA. This article reviews the progress and achievements obtained to date. The latter include the development of ELISAs employing recombinant antigen for detection of antibody to O. volvulus which have both high sensitivity and specificity, the commercial availability of immunoassays to measure circulating antigen in W. bancrofti infection and the generation of specific DNA-based detection systems for all three parasites.

Proposed panel of diagnostic criteria, including the use of ultrasound, to refine the concept of 'endemic normals' in lymphatic filariasis.

Although living adult Wuchereria bancrofti worms can be detected by ultrasound examination of the scrotal area in approximately 80% of men infected with this filarial parasite, the location of the adult worms in the remaining 20% remains unclear. To determine this, 32 individuals who had W. bancrofti microfilaraemia but no adult worms detectable on ultrasound were treated with diethylcarbarmazine (DEC), either with a single 6 mg/kg dose (n = 13) or with a 12-day course of 6 mg/kg per day (n = 19). They were then monitored with serial physical and ultrasound examinations. Thirteen (41%) subjects developed small, single scrotal nodules 12 h to 7 days after treatment; this rate was unaffected by the dose of DEC. No nodules were detected outside the scrotal area. All 5 men with lymphangiectasia suspected on ultrasound before treatment developed scrotal nodules, compared to 8 (29.6%) of 27 men without ultrasonographic evidence (P = 0.006). Thus, using both ultrasound and 'provocative' treatment with DEC, adult W. bancrofti can be detected in the scrotal area of an estimated 88% of infected men. Because no single diagnostic test for W. bancrofti infection is completely sensitive, a panel of tests, including ultrasound, is proposed to identify with greater accuracy 'endemic normals' for immunological and epidemiological studies.

Components of the vectorial capacity of Aedes poicilius for Wuchereria bancrofti in Sorsogon province, Philippines.

Components of vectorial capacity (biting density, survival, and host preferences) of a population of Aedes (Finlaya) poicilius, the principal vector of Wuchereria bancrofti in the Philippines, were studied in 1994-1995 in Sorsogon province. Aedes poicilius comprised 20.7% of 3243 mosquitoes of 24 species taken in 42 nights of human-biting collections, and 1.4% of 7586 mosquitoes of 27 species taken in 18 concurrent nights of carabao-trap collections. There were, on average, 16 bites by Ae. poicilius per person per night. As there was no relationship between body size of the female Ae. poicilius and parity status, body size was not a variable affecting survivorship of Ae. poicilius. The probability of daily survival was estimated to be 0.85, and the human blood index was 0.67. None of the 862 Ae. poicilius examined contained W. bancrofti larvae, probably because of distribution of diethylcarbamazine to microfilaraemic people prior to the study. The vectorial capacity of the Ae. poicilius population under study was estimated to be 2.4 new cases per primary case per day. However, the basic reproductive rate (i.e. the number of new cases of W. bancrofti infection generated from a single infective case) was estimated to be very low because of inefficiency in parasite transmission.

Components of the vectorial capacity ofAedes poiciliusforWuchereria bancroftiin Sorsogon province, Philippines

Components of vectorial capacity (biting density, survival, and host preferences) of a population of Aedes (Finlaya) poicilius, the principal vector of Wuchereria bancrofti in the Philippines, were studied in 1994–1995 in Sorsogon province. Aedes poicilius comprised 20.7% of 3243 mosquitoes of 24 species taken in 42 nights of human-biting collections, and 1.4% of 7586 mosquitoes of 27 species taken in 18 concurrent nights of carabao-trap collections. There were, on average, 16 bites by Ae. poicilius per person per night. As there was no relationship between body size of the female Ae. poicilius and parity status, body size was not a variable affecting survivorship of Ae. poicilius. The probability of daily survival was estimated to be 0.85, and the human blood index was 0.67. None of the 862 Ae. poicilius examined contained W. bancrofti larvae, probably because of distribution of diethylcarbamazine to microfilaraemic people prior to the study. The vectorial capacity of the Ae. poicilius population under study was estimated to be 2.4 new cases per primary case per day. However, the basic reproductive rate (i.e. the number of new cases of W. bancrofti infection generated from a single infective case) was estimated to be very low because of inefficiency in parasite transmission.

Diagnosis of Wuchereria bancrofti infection by the polymerase chain reaction using urine and day blood samples from amicrofilaraemic patients

A sensitive and specific polymerase chain reaction (PCR) based on a highly repeated deoxyribonucleic acid (DNA) sequence (188 bp; SspI repeat) was tested for the detection of Wuchereria bancrofti DNA in blood and urine samples collected during the day from individuals in Coque, Recife, Brazil, an endemic area for W. bancrofti. All microfilaraemic individuals were also positive by PCR, irrespective of the samples used. The PCR system was capable of detecting W. bancrofti DNA in amicrofilaraemic individuals: c. 93% were positive by PCR when day blood samples were used and 59·7% when urine samples collected at 07:00 were used. Thus, nocturnally periodic W. bancrofti infection can be detected in blood samples collected during the day, which is convenient for large-scale screening. In addition, non-invasive urine collection provided suitable samples for PCR, which is clearly advantageous for preliminary mass diagnosis.

Quantitative assessment of circulating antigens in human lymphatic filariasis: a field evaluation of monoclonal antibody-based ELISA using blood collected on filter strips.

To quantify circulating antigens in individuals with lymphatic filariasis by means of an ELISA using blood on filter strips. Circulating antigens in filarial patients and normal individuals living in an area endemic for W. bancrofti infection in Madras, India were estimated using a monoclonal antibody-based ELISA. All microfilaraemics showed positivity to circulating antigens whereas people with chronic pathology and 80% of the endemic normals tested negative. The antigen levels in the blood collected in the night and during day time showed positivity and there was no difference in the antigen concentration. The results of the antigen levels collected onto filter strips correlated with their corresponding plasma antigen levels (r = 0.83). In microfilaraemics, DEC treatment did not alter the levels of circulating antigens for up to one month. We conclude that this monoclonal antibody-based ELISA using filter strips may be used in day time and replace the existing routine night blood surveys in our endemic area in India.

Immunodiagnosis of human lymphatic filariasis: A review

A monoclonal antibody-based antigen detection system was used to detect the levels of circulating antigen in filarial patients before and after treatment with DEC and in normal individuals living in an area endemic forW. bancrofti infection in Chennai, India. The present study was to show the use of this assay as a means of efficient screening for filariásis in an endemic area where blood was absorbed onto the filter paper by finger prick during day time. The results of the antigen levels collected onto filter strips correlated with their corresponding plasma antigen levels (r=0.83). In microfilaraemics, DEC treatment did not alter the levels of circulating antigens upto a period of one month. We conclude that this monoclonal antibody based ELISA using filter strips may be used in daytime and can replace the existing routine night blood survey.

Our experience with experimentalWuchereria bancrofti infection in Indian langur (Presbytis entellus)

Our experience with experimentalWuchereria bancrofti infection in Indian langur (Presbytis entellus)

A polymerase chain reaction-based assay for detection of Wuchereria bancrofti in human blood and Culex pipiens

Human blood samples and indoor-resting Culex pipiens were collected in 33 randomly selected houses from different sectors of a village in the Nile Delta of Egypt which was endemic for Wuchereria bancrofti. Blood was also collected from subjects with no history of living in filarial endemic areas. Human blood samples were divided and assessed by both membrane filtration and polymerase chain reaction (PCR). Similarly, mosquito samples were assessed by both dissection and PCR. Blood pools representing each household were tested by PCR. If a pool gave a positive result, then individual blood specimens were also tested by PCR. Of the 33 houses tested, both membrane filtration and blood pools assayed by PCR identified 14 (42·4%) ‘infected houses’. PCR detected parasite deoxyribonucleic acid (DNA) in blood pools from an additional 3 households that gave negative results by membrane filtration. Of 178 endemic blood samples tested by membrane filtration, 22 (12·3%) had microfilariae and all were individually positive by PCR. Although microfilaria counts were lower in blood collected during the day than in night-collected blood, the PCR results were consistent, regardless of time of collection. All non-endemic blood samples were negative by PCR. Among the 33 houses tested, mosquito pools assayed by PCR identified 17 (51·5%) as ‘infected households’. Of these, 8 houses (47%) contained at least one microfilaraemic resident. One ‘infected household’ was identified by mosquito dissection. We concluded that PCR is a powerful epidemiological tool for screening villages for the prevalence of W. bancrofti. PCR detection of W. bancrofti DNA in blood-fed mosquitoes could be used initially to locate endemic areas with transmission of bancroftian filariasis. PCR detection of W. bancrofti DNA in blood collected during the day could then be used to assess W. bancrofti infection rates.

Reduction of Wuchereria bancrofti adult worm circulating antigen after annual treatments of diethylcarbamazine combined with ivermectin in French Polynesia.

Circulating filarial antigen (CFA), determined with Og4C3 ELISA, is a marker of Wuchereria bancrofti adult worm infection. The reduction of CFA over 2 years was determined in 185 microfilaremic and 111 amicrofilaremic but CFA+ adults given an annual dose of either diethylcarbamazine (DEC) or ivermectin or the two combined. Reduction of CFA level was good with DEC but weak with ivermectin and followed the same pattern in amicrofilaremic and microfilaremic groups. Combinations and DEC alone had a similar impact on CFA level. CFA clearance was observed in amicrofilaremic but not in microfilaremic persons in all DEC-containing treatments. However, the highest clearance rate was observed in persons treated with DEC at 6 mg/kg combined with ivermectin. Continuous reduction of CFA level after repeated treatments shows that elimination of W. bancrofti infection, monitored by CFA clearance, might be achieved within a few years with annual treatments of DEC combined with ivermectin.

Evaluation of the Og4C3 ELISA in Wuchereria bancrofti infection: infected persons with undetectable or ultra-low microfilarial densities.

The recently developed Og4C3 ELISA, which detects circulating Wuchereria bancrofti antigen, appears promising for use in epidemiological surveys, but its sensitivity is unknown in persons with ultra-low microfilarial densities. We used the Og4C3 to test the sera of 282 persons who were microfilaria-positive in 1-16 ml of blood, 18 persons who were microfilaria-negative but who had ultrasonographic or biopsy evidence of adult W. bancrofti infection, and 63 lifelong residents of a non-endemic area of Brazil. A total of 276 (97.9%) persons with detectable microfilaraemia tested positive (optical density > 0.033). At microfilarial densities of < 1, 1-30, and > 30 microfilariae per ml of blood, the sensitivity of the Og4C3 was 72.2, 97.6 and 100%, respectively (chi 2-test for trend, P < 10(-6)). The assay was positive in 66.7% of amicrofilaraemic persons with evidence of adult worm infection and in one (1.6%) of 63 residents of the non-endemic area (specificity, 98.4%). Our findings support the increasingly widespread use of the Og4C3 for field investigations and epidemiological assessments. However, the sensitivity of the assay may be low in persons who are microfilaria-negative or with densities of < 1 microfilaria per ml.

Parasite antigenemia without microfilaremia in bancroftian filariasis.

The term "endemic normal" in the context of filariasis refers to people who are amicrofilaremic and free of clinical signs or symptoms of filariasis despite regular exposure to the parasite. Some sera from endemic normals contain soluble Wuchereria bancrofti antigens that are detectable by enzyme-linked immunosorbent assay. We now report evidence that filarial antigenemia in these people is not an artifact and that it is indicative of active W. bancrofti infection. Filarial antigenemia was first detected within one month of the onset of microfilarial patency in experimentally infected primates. Human sera from antigen-positive endemic normals contained the same filarial antigens (by Western blot) as sera from people with microfilaremia. Sera from antigen-positive endemic normals also contained significantly higher levels of immunoglobulin G4 antibodies to native and recombinant filarial antigens than sera from antigen-negative controls matched for age and sex. The epidemiology of filarial antigenemia in endemic normals was studied with sera from a population-based study of filariasis in an Egyptian village with a microfilaria prevalence of 29%. Seventeen percent of endemic normals had antigenemia, and this group comprised 11% of the total village sample. Filarial antigenemia was significantly more common in endemic normals more than 30 years of age than in younger people. These results suggest that amicrofilaremic and asymptomatic W. bancrofti infections are relatively common in endemic areas. Additional studies are needed to determine the clinical significance, prognosis, and optimal management of such infections.

Ultrasonigraphic Evidence of Abnormal Lymphatic Vessels in Young Men with Adult Wuchereria Bancrofti Infection in the Scrotal Area

Purpose We determined the prevalence and magnitude of dilatation of the lymphatic vessels of the spermatic cord in men infected with Wuchereria bancrofti, which is a known major cause of hydrocele in the tropics. Materials and Methods Scrotal ultrasound was performed with a 7.5 MHz, transducer in 78 men from Recife, Brazil (endemic for filariasis) and in 15 from a nonendemic area. Results Among men from Recife the lymphatic vessels were dilated (1.3 to 15.0 mm., mean 3.8) at the location of the adult worm. Vessel diameter was not associated with hydrocele. Conclusions Lymphatic dilatation was observed in all men with ultrasonographically detectable W. bancrofti infection, even those who were asymptomatic.