Implications of low-density microfilariae carriers in Anopheles transmission areas: molecular forms of Anopheles gambiae and Anopheles funestus populations in perspective.

BACKGROUND: Previous studies have shown a general reduction in annual transmission potential (ATP) of Anopheles species after mass drug administration (MDA) in lymphatic filariasis endemic communities. Whereas results obtained from a monitoring programme after three years of MDA revealed a decrease in ATP of Anopheles funestus this was not the same for An. gambiae s.s. in Ghana. In this study, the ability of these vectors in transmitting Wuchereria bancrofti in nine lymphatic filariasis endemic communities in Gomoa District of Ghana after four rounds of MDA with ivermectin and albendazole was investigated. METHODS: After mass screening of inhabitants in these communities, twelve consenting volunteers with different intensities of microfilariae (mf) slept under partly opened mosquito nets as sources of mf blood meal. Hourly collection of mosquitoes and finger-pricked blood were taken from 21.00 to 06.00 hours the following day. For each hour, half of the mosquitoes collected were immediately killed and dissected for mf. The remaining half were maintained up to 13 days for parasite maturation. Parasitaemia and infection rates in the mosquitoes were determined by microscopy. The mosquitoes were identified by microscopy and molecular techniques. RESULTS: A total of 1,083 participants were screened and the overall parasite prevalence was 1.6% with mf intensities ranging from 0 to 59 per 100 µl and geometric mean intensity of 1.1 mf per ml of blood. Of the 564 mosquitoes collected, 350 (62.1%) were Anopheles spp., from which 310 (88.6%) were An. funestus and 32 (9.1%) An. gambiae. Six anopheline mosquitoes (1.7%) were found infected with L1, but no larva was observed in any of the mosquitoes maintained up to 13 days. Molecular studies showed all An. gambiae s.l. to be An. gambiae s.s., of which 21 (70%) were of the M molecular form. CONCLUSION: At low-level parasitaemia after 4 rounds of MDA, there was no recovery of infective stage larvae of W. bancrofti in An. funestus s.l. as well as M and S forms of An. gambiae.

Factors affecting transmission of Wuchereria bancrofti by anopheline mosquitoes. 3. Uptake and damage to ingested microfilariae by Anopheles gambiae, An. arabiensis, An. merus and An. funestus in east Africa.

Laboratory observations were made on the uptake of microfilariae (mf) of Wuchereria bancrofti by Anopheles gambiae, An. arabiensis, An. merus and An. funestus. Over host mf densities ranging from 450/ml to 1735 mf/ml neither the percentage of mosquitoes ingesting mf nor the mean number of mf per mosquito was correlated to host mf density. All mosquito species damaged mf during ingestion but the proportion harmed was independent of host mf density. The mean proportion damaged was 0.67 in An. gambiae, 0.51 in An. merus, 0.47 in An. Arabiensis and 0.26 in An. funestus. A comparison of the mean number of undamaged mf ingested and the number of larvae in the thoracic muscles revealed that not all undamaged mf were able to reach the thoracic muscles.

Survival estimates for adult Culex pipiens in the Nile Delta.

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.

Comparative densities of Wuchereria bancrofti microfilaria in paired samples of capillary and venous blood.

Concurrent finger-prick and venous blood samples were obtained from 43 patients infected with Wuchereria bancrofti. Microfilariae were counted in blood smears and on nuclepore filters. Based on the numbers of microfilariae in 20-microliters finger-prick samples, an expected (theoretical) number of microfilariae in venous samples was calculated. This expected value was compared to the actual number of microfilariae present in venous blood. The actual number of microfilariae present ranged from 0.7 to 30 times (median 3) less than expected. The exponential function y = 16.74e.17x, r2 = .69, where y = venous counts and x = capillary counts was found to accurately reflect the relationship between venous and capillary counts. Each unit (microfilaria) change in capillary blood would result in an expected 19% unit increase/decrease in venous blood. The results support strongly the theory that microfilariae are unevenly distributed in the blood system. The epidemiological implication is that persons with low or ultralow microfilaremia levels in venous blood may have a much greater pool of microfilaria available in capillaries. We believe that the present study explains, at least partially, why mosquitoes feeding on these kinds of microfilaria carriers (frequently) have a greater uptake fo microfilariae than expected.

Factors affecting transmission of Wuchereria bancrofti by anopheline mosquitoes. 2. Damage to ingested microfilariae by mosquito foregut armatures and development of filarial larvae in mosquitoes.

Microfilariae (mf) of Wuchereria bancrofti from the midgut of 639 Anopheles gambiae, 557 An. arabiensis, 117 An. melas and 9 An. funestus were examined immediately after the mosquitoes had fed on carriers with different densities of mf. The percentages of mf damaged during ingestion were 57.1-60.0 in An. gambiae, 33.3-50.6 in An. arabiensis and 38.7-55.7 in An. melas. In each species the percentage of mf damaged was independent of mf density in the human host. A further 3657 An. gambiae, 2875 An. arabiensis, 347 An. melas and 32 An. funestus were examined 7 d or more after feeding on mf carriers. In An. gambiae and An. arabiensis, mean numbers of larvae per mosquito were strongly correlated to mf blood density, with similar regression slopes to those obtained from the regression of mf blood density on mean uptake of mf/mosquito. The ratio of mean numbers of larvae per mosquito to mean numbers of intact mf ingested per mosquito increased as the density of mf in the human host increased in An. gambiae and An. arabiensis, but decreased in An. melas as host mf density increased.

The Wellcome Trust lecture. Inflammatory responses to filarial connective tissue parasites.

The inflammatory responses to lymphatic filariae and to Onchocerca volvulus are reviewed with particular attention to evolutionary biology; inflammatory host spectrum; non-specific components; immunoregulation; immune evasion versus immunomodulation; chronic tissue damage and scarring and disease models. Basic principles of pathogenesis are emphasized, comparisons drawn with schistosome infection, and critical items of missing information are highlighted.

The examination--time/dose interval in the provocation of nocturnally periodic microfilariae of Wuchereria bancrofti with diethylcarbamazine and the practical uses of the test.

The administration of diethylcarbamazine (DEC) in the daytime in areas where microfilariae (mf) of Wuchereria bancrofti and Brugia malayi exhibit nocturnal periodicity provokes mf to enter the peripheral blood. The importance of relating the dose of DEC and time when the blood specimen is taken is emphasized. The main practical uses of this provocative day test are as a routine diagnostic tool, in estimating prevalence and density of mf in a community, as a preliminary screening method for assessing potential filaricides nd in evaluating the parasitological response to mass chemotherapy.

The mechanisms which affect the periodic cycle of Pacific Wuchereria bancrofti microfilariae.

1. Investigations were made of the effect of various procedures in raising or lowering the microfilaria count of Pacific type Wuchereria bancrofti in the peripheral blood. 2. Raising the body temperature in the early morning was followed by a moderate fall in the counts. Breathing increased oxygen, or reduced oxygen (hypoxia) or increased carbon dioxide, or the ingestion of sodium bicarbonate produced no consistent and significant change in the count. Ingestion of glucose (in one volunteer) was followed by a small rise in the count. Muscular exercise was followed by a fall in the count, which is interpreted as probably being a response to a lower concentration of oxygen in the venous blood returning to the lung. 3. It has not been possible to identify the physiological components on the circadian rhythm of the human body which entrain the cycle of these microfilariae. Attempts to obtain evidence incriminating the stimuli described above have been unsuccessful.

Tests of susceptibility of Liberian Culex quinquefasciatus to Tanzanian Wuchereria bancrofti.

Culex quinquefasciatus strains of Liberian and Tanzanian origin were fed on Tanzanian donors with Wuchereria bancrofti microfilaraemias. 11 to 17 days later the mosquitoes were dissected and scored for the presence and numbers of immature and mature filarial larvae. In one experiment a significantly lower susceptibility was found in a Liberian compared with a Tanzanian strain, but in the other cases there was no significant difference. The susceptibility of the Liberian strains was higher than in published data on the same mosquito strains tested with Liberian W. bancrofti. It is doubtful whether the Liberian mosquito strains could be a useful source of genes for the construction of a refractory strain with which to replace an East African vector population.

The susceptibility of Liberian Culex quinquefasciatus to Wuchereria bancrofti in Sri Lanka.

Strains of Culex quinquefasciatus collected in Liberia showed low susceptibility to Wuchereria bancrofti from Liberian donors. However, when the same mosquito strains were fed on W. bancrofti donors from Sri Lanka they showed equally high susceptibility as Sri Lankan C. quinquefasciatus simultaneously fed on the same donors. A Liberian strain did not respond to selection for refractoriness. Thus the strains of W. bancrofti from Liberia and Sri Lanka differ in their ability to infect specific mosquito strains and it is concluded that Liberian C. quinquefasciatus could not provide genes for use in the construction of a refractory strain intended for the replacement of Sri Lankan vector populations.

[Studies of the numerical distribution of microfilariae in foci of lymphatic filariasis (author's transl)]. / Etude de la distribution des numérations microfilariennes dans les foyers de filariose lymphatique.

The frequency distribution of Wuchereria bancrofti microfilariae counts in capillary blood, at a community level, is shown to differ significantly from the log-normal distribution, commonly used (and recommended by WHO). A much better fit is obtained with a truncated negative-binomial distribution, the k exponent of which is estimated to 0.3. This value does not seem to be affected by the endemic level, by microfilaricide mass-treatments or by the blood sample volume. This result has immediate practical interests: 1) it facilitates a posteriori comparisons between surveys in which different volumes of blood were examined; 2) at a given density it allows calculation the change in sensitivity (detection rate) as a function of a change in the volume of blood examined, thus allowing better a priori planning of surveys. The paper provides the formulae required to calculate k, or, given k, to calculate the expected numbers in the various density classes. It also provides nomograms (fig. 4a & b) giving estimates of the true mean M and of the detection rate TD, given any one of the three following observations, made on observed positives: the median Dmf. 50, the mean m, or the percentage Pl having only 1 mf. The effect of changing the volume of blood on the rate of detection can then be read easily from the nomogram. These results, likely to apply to other important parasitic diseases, should instigate more elaborated statistical research in this direction.