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Crude antigenic preparations from Setaria equina were used in ELISA and Western blotting to examine cross-reaction with human sera from areas endemic for bancroftian filariasis. Sera from normal subjects from non-endemic areas were included as negative controls. Cross-reaction was found between S. equina antigens and antibodies in the sera of Wuchereria bancrofti-infected patients, with the highest levels observed between sera of chronic infected patients and Setaria spp. crude female worm surface antigen (CFSWA). In the absence of active transmission of Setaria spp. infection, CFWSA is useful to detect chronic W. bancrofti infection before patients become symptomatic, particularly when chronic patients are known to be amicrofilaraemic. In the presence of active S. equina infection, antigens from the adult and microfilaraemic stages showed the highest degree of cross-reaction with human sera.

Human IgG antibody responses to Wuchereria bancrofti third stage infective larvae (L3) surface and somatic antigens were studied by indirect immunofluorescence (IFA) and immunoblot with endemic Egyptian sera (n = 115) with the aim of identifying targets of protective immunity. Human sera variably recognized 14 major bands in L3 by immunoblot. The statistical significance of group differences in antibody prevalence was assessed by the chi-squared test. Children and young adults (aged 10-20 years) tended to have antibodies to more L3 somatic antigens than older adults, with significant differences for bands at 66, 60 and 5 kDa. Infected subjects had more consistent antibody responses to antigens at 55, 50 and 6 kDa than endemic normal subjects with negative serum filarial antigen tests, who are presumed to be uninfected. A 5 kDa antigen was preferentially recognized by the latter group. Antibodies to L3 surface antigens were equally prevalent in uninfected children (75%) and adults (90%) but less prevalent in people with microfilaremia (38%) than in amicrofilaremic subjects with or without filarial antigenemia (81%) (P < 0.001). IFA-positive sera showed significantly enhanced recognition of antigens at 66, 40 and 14 kDa in immunoblots relative to IFA-negative sera. Additional studies are needed to further characterize antigens identified in this study and to establish whether they are indeed targets of protective immunity in humans.

Age-specific levels of Wuchereria bancrofti infection determined by microfilaremia in filarious 10 to 60 years old males from endemic village revealed that microfilaria intensity was higher in younger age, presumably because of increased vector biting rate of younger individuals. Out of 1334 Cx. pipiens captured while attempting to bite 51 child-adult bait pairs, 54.0% selected a host in the 10-15 age group. Biting preference towards children was more evaluated (79.2%) later than earlier in the night. Frequency distribution of age-specific biting collections indicated that 62.7% recorded exceeding numbers of bites on children, 21.6% adult biting, and 15.7% equal biting of one of the baits in the pair. Furthermore, 21.6% of the children were exposed to nightly bites ranging from 21 to 64 and only 7.8% of the adults experienced from 21 to 85 bites. Mosquito biting intensity averaged 14.2 and 11.6 bites/night for children and adults respectively. Children experienced 22.4% more bites per night than adults. Thus, children are more exposed to filaria infection and seem to represent an important source of infection. It is concluded that attempts at widespread control of filariasis in the densely populated Nile Delta, where youngsters may account for an immense proportion of the local population, should concentrate on immatures.

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 rested, 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.

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.

Studies were conducted to compare early phenomena associated with W. bancrofti infection and further development in the filaria vector Culex pipiens and the refractory mosquito Aedes caspius. Ingestion rates evaluated immediately after simultaneous feeding on an infected human were 69.1% for 43 Cx. pipiens and 35.7% for 28 Ae. caspius. The observed number of mf ingested by either mosquito did not vary significantly (2.7 +/- 1.4, and 2.3 +/- 0.9, respectively) and, based on the size of the blood meal ingested (2.8 and 2.3 microliters respectively), a two-fold mf concentration factor was recorded for both species. Blood ingested by Cx. pipiens (N = 16) and Ae. caspius (N = 10) clotted within 120 and 90 min post-feeding respectively. The time difference observed, however, did not affect significantly the rates of migration into the hemocele (56 and 67% respectively). Comparison of initial infection rates with those obtained after the extrinsic incubation period of the parasite was completed, indicated that the proportion of infected Cx. pipiens was reduced by 3.9% and that of Ae. caspius by 30.0%. Furthermore, the observed infectivity ratio of 265 Cx. pipiens that had an infective blood meal was 0.74 and only 0.009 for 70 Ae. caspius. It is concluded that refractoriness of Ae. caspius to W. bancrofti is expressed through the feeding mechanism itself, by severely limiting the mf ingestion rate, and through physiological processes that inhibited the development of ingested worms.

We compared the defense mechanisms directed against Wuchereria bancrofti by a filaria susceptible mosquito, Culex pipiens, and a refractory one, Aedes caspius. The reciprocal deleterious effects of both the mosquito host and its parasite are reported. Anatomical and histological examinations of mosquitoes revealed minor differences in the general structure of the foregut of either species. The cibarial pump of Ae. caspius, however, lacked a cibarial armature and damage inflicted to some of the microfilariae (mf) ingested by this mosquito was not mechanical. In contrast, the cibarial armature of Cx. pipiens consisted of 24 delicate teeth which did not seem to affect ingested mf. The peritrophic membrane (pm) did not constitute a significant barrier to mf migrating through the midgut wall of either mosquito. Indeed, mf forced their way out into the hemocoel, immediately after they have been ingested, and up to 60 min post-feeding, when the pm was still thin and soft. It hardened 16 h post-feeding. Traversing mf perforated the pm and displaced the basement membrane. Within the thorax of Cx. pipiens, the parasite completed its development synchronously. In contrast, mf ingested by Ae. caspius did not develop beyond the sausage stage, and many of these larvae were vacuolated. In both mosquitoes, parasitized thoracic muscle fibers were tunnelled and, only in Ae. caspius, their nuclei were pyknotic. No worm encapsulation or melanization was observed in either mosquito. It is concluded that defense lines to invading parasites in Cx. pipiens are insignificant, and that in Ae. caspius, the main barrier to parasite development resides within the thoracic muscles. In the absence of cellular evidence, it is suggested that such a barrier is physiological in nature.

Studies were conducted in the Nile Delta of Egypt to determine the feasibility of detecting Wuchereria bancrofti microfilariae (Mf) in mosquitoes as a primary surveillance method for the identification of filariasis-endemic villages. Initial experimental studies evaluated the ingestion, survival, and migration rates of W. bancrofti Mf in Culex pipiens and Culex antennatus after mosquitoes were fed on infected volunteers. In 2 villages, 1,684 bloodfed mosquitoes were dissected during the night immediately after collections inside houses. In the village of Kafr Tahoria, Mf were found in 27 of 519 Cx. pipiens and in one of 8 Anopheles pharoensis. In Tahoria, Mf were detected in 7 of 799 Cx. pipiens and in one of 302 Cx. antennatus. Identifying filariasis-endemic villages based on the detection of Mf in mosquitoes may be a useful strategy for epidemiologic studies or for filariasis control programs.

To demonstrate focality of filariasis within endemic rural areas and to define exposure variables which may influence this phenomenon, the population of an agrarian endemic village, of 12,500 individuals, in the Nile Delta of Egypt was censused. A sequential sample of individuals residing in every fifth house was tested for microfilaremia (239 households with 8.6 +/- 3.5 individuals per household (HHD). Three areas of the village were tested simultaneously and a questionnaire was filled out for each sampled HHD with special emphasis given to the entomological and environmental factors that might affect filarial infection. One area (area A) had a higher intensity of larvae and biting adults of the main filarial vector, Culex pipiens, than the other two areas (areas B and C). Of the 1488 persons who agreed to be tested in the three areas 181 (12.2%) were microfilaremic. Microfilaremia prevalences were the same in males and females and microfilariae were present in all age groups. Filarial infection was most prevalent in area "A" (1.16 +/- 0.14 infected people per HHD) than in area "B" (0.44 +/- 0.11) or "C" (0.72 +/- 0.10) (ANOVA; p = 0.0003). several possible predictor variables were analyzed by logistic regression with the presence of infection as the response variable. Among individuals residing around the main Cx. pipiens development sites, those living in houses facing vacant land are exposed to more mosquito bites and had a greater chance of having filarial infection (relative risk [RR] = 1.5; logistic regression, P = 0.0089). People residing in large households had a reduced chance of having filarial infection (RR = 0.87; logistic regression, p = 0.0015). These data show that the distribution of microfilaremic individuals is uneven within the study village and suggest that small HHD and houses that bordered open areas containing mosquito development sites are potential risk factors for acquiring filarial infection.

Females of Culex pipiens L. experimentally infected with Wuchereria bancrofti were tested for in vitro transmission of infective stage L3 larvae using a capillary tube technique. Of the females harbouring L3 larvae, 60.7% (34/56) transmitted a geometric mean of 2.84 L3 larvae (range 1-35). Transmitting Cx. pipiens contained significantly more L3 larvae than non-transmitters. Both the probability of transmission and the number of L3 larvae transmitted were related to the total number of loads larvae/mosquito. Almost all females with L3 loads > or = 10 transmitted larvae in vitro. The possibility is discussed that the detection of L3 larvae in field collected mosquitoes by dissection may overestimate the proportion capable of parasite transmission and the proportion of L3 larvae deposited on host skin by Ca. 40%.

We have previously reported that a monoclonal antibody-based antigen detection assay (AD12) is sensitive and specific for Bancroftian filariasis in Egypt. The purpose of the present study was to demonstrate the use of this assay in a sentinel population as a means of efficiently screening for filariasis endemicity. Antigen testing was performed with finger-prick blood collected during the day from 743 schoolchildren (ages 11-16 years). The school draws students from 5 villages in Qalubia Governorate, 35 km north-east of Cairo, Egypt. The prevalence of filarial antigenaemia in the school was 17.2%. Antigenaemia rates in children from the 5 villages were 29, 20, 18, 17, and 10% (non-uniformity significant by chi 2 analysis, P = 0.02). These data agree with Ministry of Health rankings of relative endemicity for these villages based on prior night blood surveys. The village with the highest antigen prevalence in children was surveyed one year before the present study. Prevalence rates of antigenaemia and microfilaraemia at that time for a different sample of children aged 11-16 years were 33% and 22%, respectively. We conclude that antigen detection in schoolchildren of this age group is an efficient means of assessing filariasis endemicity in Egypt.

The objective of this study was to find the best tests for efficiently estimating the true prevalence of Bancroftian filariasis in endemic areas. The study population comprised 427 people over 10 years of age in an endemic village in Egypt. Four tests were evaluated; a standardized clinical examination, night blood examinations for microfilariae (50 microL thick films and 1 mL membrane filtration), and a test for circulating filarial antigen. 191 subjects (44.75%) had at least one positive test and were considered to have filariasis. The sensitivities of clinical examination, thick films, membrane filtration and antigen testing for filariasis were 16%, 50%, 64%, and 88%, respectively. Relative to membrane filtration of night blood, the filarial antigen test had a sensitivity of 97.5%, a positive predictive power of 71%, and a negative predictive power of 99%. None of the blood tests was a sensitive indicator of clinical filariasis; 69% of clinical cases were negative in all 3 blood tests and would have been missed if clinical examinations had not been done. Therefore, we recommend a combination of clinical examination and the filarial antigen test (with optional examination for microfilariae of those with positive antigen tests) for community diagnosis of Bancroftian filariasis in endemic areas.

Conventional methods for diagnosis of Wuchereria bancrofti infection are insensitive and often impractical because of the need for night blood collections. A sensitive and specific antigen detection assay has been developed for W. bancrofti, which is based on a monoclonal antibody (AD12) that binds to a repeated epitope on a 200 kDa adult worm excretion product present in sera from infected humans. The only formal evaluation of this assay to date was performed with sera from India. In the present study, we have evaluated the performance of the AD12 antigen assay in two laboratories with sera collected in endemic and non-endemic areas in Egypt. Antigen was detected in 57 of 59 (97%) sera from microfilaremic subjects, and in 22 of 139 asymptomatic and amicrofilaremic subjects who reside in a highly endemic area. Antigen titers were significantly correlated with microfilaria counts (r = 0.41, P less than 0.01). Filarial antigen was not detected in most sera from amicrofilaremic subjects with clinical filariasis. Comparative antigen test results obtained from laboratories in Cairo and St. Louis agreed in 170 of 173 sera tested. Filarial antigen was not detected in sera from Egyptians with no history of residence in filaria-endemic areas. Specifically, nonendemic sera from patients with other parasitic infections (schistosomiasis, fascioliasis, ascariasis, etc.) were uniformly negative in the assay. We conclude that the AD12 filarial antigen assay is sensitive and specific for W. bancrofti infection in Egypt.

In an endemic area for Wuchereria bancrofti filariasis in the Nile Delta, survival of adult female Culex pipiens was estimated by parity rate, mosquito infection and infectivity rates. Infection rates and 4th instar larval populations, as well as infection and parity rates, were linearly correlated. Infectivity correlated only with parity rates. These associations corresponded to parallel changes in ambient temperature. Although survival calculated from parity rates measured longevity of both infected and non-infected Cx. pipiens, survival based on infection and infectivity was a more reliable indicator for parasite transmission.