Factors influencing silhouette trap captures of the blackfly Austrosimulium bancrofti in Queensland, Australia

Three silhouette traps of novel design were used to investigate the trap finding ability of female Austrosimulium bancrofti (Taylor) (Diptera: Simuliidae) in southeast Queensland. The traps were 'upright', 'elongate' or 'square' in shape. Five highly significant factors: hour after sunrise, temperature, relative humidity, trap shape and distance between traps, were included in a model which generated fitted estimates of flies captured under defined conditions. The rate of increase and the level of maximum capture rate indicated that the elongate trap captured the highest numbers of blackflies. An interaction between hour after sunrise and temperature suggests a strong endogenous modification of trap finding activity. Numbers captured increased with increasing distance between traps and decreasing relative humidity. The importance of these results in understanding behaviour of A. bancrofti is discussed.     

Transmission dynamics of lymphatic filariasis: density-dependence in the uptake of Wuchereria bancrofti microfilariae by vector mosquitoes

Gaining a better understanding of parasite infection dynamics in the vector mosquito (Diptera: Culicidae) population is central to improving knowledge regarding the transmission, persistence and hence control of lymphatic filariasis. Here, we use data on mosquito feeding experiments collated from the published literature to examine the available evidence regarding the functional form of the first component of this parasite-vector relationship for Wuchereria bancrofti (Filarioidea: Onchocercidae) causing Bancroftian filariasis, i.e. the rate of microfilariae (mf) uptake from the blood of infected humans by the feeding mosquito vector. Using a simple logarithmic regression model for describing the observed relationships between the mean numbers of mf ingested per mosquito and parasite load in humans in each study, and a linear mixed-effects meta-analytical framework for synthesizing the observed regressions across studies, we show here for the first time clear evidence for the existence of density-dependence in this process for all the three major filariasis transmitting mosquito vectors. An important finding of this study is that this regulation of mf uptake also varies significantly between the vector genera, being weakest in Culex, comparatively stronger in Aedes and most severe and occurring at significantly lower human mf loads in Anopheles mosquitoes. The analysis of the corresponding mf uptake prevalence data has further highlighted how density-dependence in mf uptake may influence the observed distributions of mf in vector populations. These results show that whereas strong regulation of mf uptake, especially when it leads to saturation in uptake at low human parasite intensities, can lead to static distributions of mf per mosquito with host parasite intensity, a weaker regulation of mf ingestion can give rise to changes in both mean mf loads and in the frequency distribution of parasites/mosquito with increasing human parasite intensity. These findings highlight the importance of considering local vector infection dynamics when attempting to predict the impacts of community-based filariasis control. They also emphasize the value of developing and applying robust meta-analytic methods for estimating functional relationships regarding parasitic infection from population ecological data.     

Haemocyte population and ultrastructural changes during the immune response of the mosquito Culex quinquefasciatus to microfilariae of Wuchereria bancrofti

Abstract Haemocytes circulating in the haemolymph protect insects against pathogens that enter the haemocoel. Changes in haemocyte morphology and differences in haemocyte counts during the immune response of Culex quinquefasciatus Say (Diptera: Culicidae) to microfilariae of Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae) were investigated in the present study. The mean number of total haemocytes was significantly elevated in infected mosquitoes (P < 0.001), reaching a peak on the third day post‐infection. Differential counts show that mean numbers of prohaemocytes, plasmatocytes, granular cells and oenocytoids increased significantly after infection with microfilariae granulocytes compared to the control and näive groups of Cx. quinquefasciatus (P < 0.05). Changes in proportional counts of haemocytes were also analysed in haemolymph perfusates of Cx. quinquefasciatus infected with W. bancrofti. On the first day post‐infection, infected mosquitoes showed an increase in the proportion of prohaemocytes (18.8% compared to 9.6% for the control) and of oenocytoids (7.1% compared to 4.7% control); however, they exhibited lower levels of plasmatocytes (36.6% compared to 42.1% control) and granular cells (36.1% compared to 41.4% control). On day 14 post‐infection, similar changes were observed for these haemocyte types, except that the proportion of granular cells was significantly greater than the control (41.2% compared to 31.3% control). Although an enhancement of prohaemocyte numbers was observed, this cellular type did not show any ultrastructural alteration. On the other hand, granular cells, plasmatocytes and oenocytoids presented morphological alterations indicative of innate immunological activation in mosquitoes infected with W. bancrofti.     

Ambient temperature effects on the extrinsic incubation period of Wuchereria bancrofti in Aedes polynesiensis: implications for filariasis transmission dynamics and distribution in French Polynesia.

Temperature effects on development of the human filarial parasite Wuchereria bancrofti (Cobbold) (Filaridea: Onchocercidae) in the main Pacific vector Aedes polynesiensis Marks (Diptera: Culicidae) are analysed in relation to ambient climatic conditions. A statistical model of the extrinsic cycle duration as a function of temperature is described and used to distinguish three patterns of W. bancrofti transmission dynamics: continuous, fluctuating and discontinuous, occurring from north to south geographically among French Polynesian archipelagos. In the northerly Marquesas Islands (8-11 degrees S) filariasis transmission is continuous and very active, facilitated by perennially high temperatures combined with constantly high rates of man-vector contact. In the southerly Australes Islands (21-28 degrees S) filariasis transmission is seasonally discontinuous and, during the cooler months (May-September), the model predicts virtually no transmission because the cycle duration exceeds the life expectancy of the vector. In the Society Islands (16-18 degrees S), between the Marquesas and Australes, transmission is predicted to be intermediate as expected from their latitude, with seasonally fluctuating transmission potential. In the Tuamotu Islands (also geographically intermediate: 14-23 degrees S), with theoretically perennial transmission potential, transmission occurs only intermittently, being limited by other human and environmental factors whereby man-vector contact is confined to seasonal agricultural situations. Generally, among French Polynesian archipelagos where Aedes polynesiensis is the vector, the transmission potential for W. bancrofti and resulting disease manifestations of lymphatic filariasis in humans are correlated with ambient temperature due to the degree of southern latitude.     

Assessing density dependence in the transmission of lymphatic filariasis: uptake and development of Wuchereria bancrofti microfilariae in the vector mosquitoes

Understanding density dependence in the transmission of lymphatic filariasis is essential for assessing the prospects of elimination. This study seeks to quantify the relationship between microfilaria (Mf) density in human blood and the number of third stage (L3) larvae developing in the mosquito vectors Aedes polynesiensis Marks and Culex quinquefasciatus Say (Diptera: Culicidae) after blood-feeding. Two types of curves are fitted to previously published data. Fitting a linearized power curve through the data allows for correction for measurement error in human Mf counts. Ignoring measurement error leads to overestimation of the strength of density dependence; the degree of overestimation depends on the accuracy of measurement of Mf density. For use in mathematical models of transmission of lymphatic filariasis, a hyperbolic saturating function is preferable. This curve explicitly estimates the Mf uptake and development at lowest Mf densities and the average maximum number of L3 that can develop in mosquitoes. This maximum was estimated at 23 and 4 for Ae. polynesiensis and Cx. quinquefasciatus, respectively.     

Analysis of polymorphism of 18S rRNA gene in Wuchereria bancrofti microfilariae

The polymorphism of the 18S rRNA gene in Wuchereria bancrofti microfilariae (mf) collected from three different zones in India was analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The RFLPs of the amplified products obtained after digestion with restriction enzymes Ssp I, Msp I and Hha I showed no difference in the banding patterns among the mf isolates from different endemic zones. Further the sequencing of PCR products did not show any difference in the nucleotide sequence either. The phylogenetic analysis of the sequences of W. bancrofti mf isolates from different endemic zones has shown branching with the earlier reported sequences of W. bancrofti and its close relative Brugia malayi.     

Rhythmic Behaviour of W. Bancrofti Microfilaraemia in Human Population at Raipur

The temporal dynamics of microfilariae (mf) in human blood, and the biting activity and mf density in the vector were studied. Finger-prick blood samples (in duplicate) were collected from 14 microfilaraemic subjects at two-hourly intervals over a single 24-hour timescale. Samples were taken from 39 subjects at 22:00 in the first week of every month for a period of 17 consecutive months. The blood slides were dried, stained and examined for presence of microfilariae. Data were analyzed with cosinor rhythmometry. A statistically validated circadian rhythm was detected in mf density in 86% of the microfilaraemic subjects. For the circannual rhythm, the peak timing for microfilariae density was recorded in July, although the spread of the peak between subjects was quite wide, extending between May 17 and September 07. In a separate study, temporal variations in biting activity, mf density in the vector and in human blood were studied. The dynamics of parasite transmission were found to be sequential, with the peaks in the circadian rhythms appearing in the order: mf in human blood, biting activity and mf density in the vector. The biting periodicity appeared to be much more stable in all subjects; the peak appeared at 02:37 with a range between 00:39 and 03:22. The peak in mf frequency was earlier by 3 hours and 25 minutes, whereas the peak in mf frequency in the vector was later by about 2 hours 4 minutes. The observed synchronization between the rhythms of vector and host appears to be essential for effective transmission of diseases. Therefore, the application of chronobiologic principles may have a bearing in the success and failure of any future strategies aimed at the prevention, cure and management of filariasis.     

斑氏丝虫感染Scid小鼠模型的建立

采用人工感染法,对严重联合免疫缺陷小鼠(Scid 小鼠) 进行夜现周期性斑氏丝虫实验感染研究。结果表明,斑氏丝虫经人工感染后能在Scid 小鼠体内发育成熟并产生微丝蚴,且虫体回收率较高为6～30% ,平均14-8% 。感染的10 只小鼠均在外周血中检获出微丝蚴,及解剖收集到成虫,感染成功率高达100% 。作者认为Scid 小鼠由于T、B淋巴细胞功能缺如,不能产生相应的抗丝虫感染抗体,是一种易于感染成功、潜伏期短、微丝蚴密度及成虫回收率高的较理想的实验动物。     

Development and evaluation of a rapid flow-through immuno filtration test using recombinant filarial antigen for diagnosis of brugian and bancroftian filariasis

There is an imperative need to develop a rapid antibody test that can be used for diagnosis of clinical cases in travelers and expatriates, primary surveillance in areas of unknown endemicity, detection of early infection in childhood and for monitoring chemotherapeutic programs. A rapid-format, simple and qualitative flow through immuno filtration test has been developed for the identification of total IgG antibodies to recombinant filarial antigen WbSXP-1. This test system employs colloidal gold-protein A reagent as the antibody capture reagent. The sensitivity and specificity of the test was evaluated in a total of 1,230 serum samples. The sensitivity of the test was found to be 90.8% with brugian (n = 70) and 91.4% with bancroftian (n = 140) microfilaraemic subjects. The test showed minimum reactivity (4/10) with Loa loa microfilaria (MF) positive sera and no reactivity (0/20) with Onchocerca MF positive sera. This rapid diagnosis is found to be non-reactive with individuals having other parasitic diseases including schistosomiasis (n = 10), soil-transmitted helminthiases (n = 34) and protozoan infections (n = 33) indicating the potential of this test as a prospective method of diagnosis for both brugian and bancroftian lymphatic filariasis. Stability kinetics was studied at different temperatures and different time periods. The rapid flow-through immuno filtration test is advantageous since it can be stored at room temperature, is user friendly and is particularly applicable in the field as an initial screening method, for epidemiological monitoring of filarial infections in bancroftian and brugian endemic regions of the world.     

Transmission dynamics of lymphatic filariasis: vector-specific density dependence in the development of Wuchereria bancrofti infective larvae in mosquitoes

The principles of meta-analysis developed in a previous study were extended to investigate the process of Wuchereria bancrofti (Cobbold) (Filarioidea: Onchocercidae) infection in mosquito (Diptera: Culicidae) hosts, focusing specifically on the functional forms and strength of density dependence in the development of ingested microfilariae (mf) to infective (third instar) larvae (L3). Mathematical models describing observed mf-L3 functional responses for each of the major three parasite-transmitting vector genera, Aedes, Culex and Anopheles mosquitoes, were fitted to paired mf-L3 data collated from all available studies in the published literature. Model parameters were estimated and compared by deriving and applying a data synthetic framework, based on applying a non-linear weighted regression model for fitting mathematical models to multistudy data. The results confirm previous findings of the existence of significant between-genera differences in the mf-L3 development relationship, particularly with regard to the occurrence of limitation in Culex mosquitoes and facilitation in Aedes and Anopheles mosquitoes. New and unexpected findings regarding L3 development from ingested mf were discovered as follows: (1) for Culex, overcompensation in L3 development at higher intensities of mf (or a peaked mf-L3 functional response) was detected; (2) for Aedes mosquitoes, facilitation (with an apparent asymptotic constraint on L3 development at high mf densities) was shown to be the major process governing L3 development, and (3) for Anopheles, a stronger facilitation type of response with no apparent saturation in L3 development appears to govern L3 output from ingested mf. These results yield major new insights regarding filarial vector infection dynamics and their potential impacts on parasite control, and demonstrate the efficacy of employing a data synthetic approach to reveal and estimate parasitic infection processes in host populations.