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Background: The time necessary for malaria parasite to re-appear in the blood following treatment (re-infection time) is an indirect method for evaluating the immune defences operating against pre-erythrocytic and early erythrocytic malaria stages. Few longitudinal data are available in populations in whom malaria transmission level had also been measured., Methods: One hundred and ten individuals from the village of Ndiop (Senegal), aged between one and 72 years, were cured of malaria by quinine (25 mg/day oral Quinimax in three equal daily doses, for seven days). Thereafter, thick blood films were examined to detect the reappearance of Plasmodium falciparum every week, for 11 weeks after treatment. Malaria transmission was simultaneously measured weekly by night collection of biting mosquitoes., Results: Malaria transmission was on average 15.3 infective bites per person during the 77 days follow up. The median reappearance time for the whole study population was 46.8 days, whereas individuals would have received an average one infective bite every 5 days. At the end of the follow-up, after 77 days, 103 of the 110 individuals (93.6%; CI 95% [89.0-98.2]) had been re-infected with P. falciparum. The median reappearance time ('re-positivation') was longer in subjects with patent parasitaemia at enrolment than in parasitologically-negative individuals (58 days vs. 45.9; p = 0.03) and in adults > 30 years than in younger subjects (58.6 days vs. 42.7; p = 0.0002). In a multivariate Cox PH model controlling for the sickle cell trait, G6PD deficiency and the type of habitat, the presence of parasitaemia at enrolment and age >/= 30 years were independently predictive of a reduced risk of re-infection (PH = 0.5 [95% CI: 0.3-0.9] and 0.4; [95% CI: 0.2-0.6] respectively)., Conclusion: Results indicate the existence of a substantial resistance to sporozoites inoculations, but which was ultimately overcome in almost every individual after 2 1/2 months of natural challenges. Such a study design and the results obtained suggest that, despite a small sample size, this approach can contribute to assess the impact of intervention methods, such as the efficacy vector-control measures or of malaria pre-erythrocytic stages vaccines.

Background & Objectives: Investigations were conducted to obtain key entomological indices of Anopheles gambiae s.l. at Igbo-Ora, a rural community in south-west Nigeria., Methods: Mosquitoes were caught daily for a week from rooms where tenants had slept the previous night in each of the four months June, July (2001), and August, September (2002). Anopheles gambiae s.l. sibling species were PCR-identified, the blood meal origin was determined by direct ELISA, and the circumsporozoite antigen by sandwich ELISA. Mean weekly rates were calculated., Results: The mean human biting rates were 0.90 and 1.6 in 2001 and 2002 respectively. The mean weekly anthropophilic rates for An. gambiae s.l. were 82 and 86% in 2001 and 2002 respectively; they were high in An. gambiae s.s., An. arabiensis and non-identified species in the complex. The mean weekly circumsporozoite rates were 6.70% in 2001 and 6.30% in 2002. The mean weekly entomological inoculation rates (EIR) were 4.95 and 5.05 in 2001 and 2002 respectively; the seasonal (6-month) rates were high: 128.7 in 2001 and 131.3 in 2002, compared to data from other rural communities on the continent., Interpretation & Conclusion: The implications of these findings on the role of An. gambiae s.l. in the holoendemicity of malaria at Igbo-Ora are discussed.

Introduction: Although cases of leishmaniasis have been reported in the province of Guaviare, Colombia, no entomological studies were included to identify the Lutzomyia sand fly vector species in that area., Objective: Lutzomyia species were identified from four townships of Guaviare. Probable vectors were named based on those species involved in transmission in other areas., Materials and Methods: Sampling was undertaken with CDC light traps suspended at heights between 1.5 m and 9 m. Additional sand flies were collected with Shannon traps and by aspiration of adult flies from daytime resting sites., Results: Sand flies belonging to 37 different species were collected. 35 of them were recorded for the first time in Guaviare Province. Four species were new records for Colombia: Lutzomyia begonae, L. campbelli, L. sericea and L. nematoducta. The most abundant species were L. hirsuta 24.3% (148/610), L. yuilli 15.2% (93/610), L. davisi 10.3% (63/610), followed by L. fartigi, L. carrerai, L. antunesi, L. flaviscutellata and L. olmeca bicolor., Conclusion: Seven of these species of have been associated previously with endemic or epidemic transmission of leishmaniasis.

Plasmodium falciparum malaria is a serious tropical disease that causes more than one million deaths each year, most of them in Africa. It is transmitted by a range of Anopheles mosquitoes and the risk of disease varies greatly across the continent. The "entomological inoculation rate" is the commonly-used measure of the intensity of malaria transmission, yet the methods used are currently not standardized, nor do they take the ecological, demographic, and socioeconomic differences across populations into account. To better understand the multiplicity of malaria transmission, this study examines the distribution of transmission intensity across sub-Saharan Africa, reviews the range of methods used, and explores ecological parameters in selected locations. It builds on an extensive geo-referenced database and uses geographical information systems to highlight transmission patterns, knowledge gaps, trends and changes in methodologies over time, and key differences between land use, population density, climate, and the main mosquito species. The aim is to improve the methods of measuring malaria transmission, to help develop the way forward so that we can better assess the impact of the large-scale intervention programmes, and rapid demographic and environmental change taking place across Africa.

Background: The current control strategy adopted by African Programme for Onchocerciasis Control (APOC) is the mass distribution of ivermectin in all endemic areas for between 12-15 years. One of the major obstacles of the strategy, is the ability of the fly to migrate long distances and this brings about the possibility of flies migrating from uncontrolled areas to controlled areas., Objective: To assess the impact of annual ivermectin treatment of onchocerciasis and to ascertain the seasonality of transmission and its impact on the current control strategy in the forest zone around a dam in Southern Nigeria., Methods: A 36 x 11-hour entomological survey was carried out in the study area. Simulium damnosum s.l were caught using two human baits from 07.00 hours-18.00 hours, three days of every month for 12 months according to the method of Walsh et al. The head, thorax and abdomen of each insect were dissected separately. Catches were made from dawn to dusk., Results: Catches yielded a total of 2,807 female S. damnosum s.l. Higher mean biting rates (2,486 flies/man/year) occurred during the dry season. The mean monthly distribution of parous flies also showed distinct seasonality with preponderance of parous flies during the dry season (61.8%) as compared to the rainy season (38.6%). Flies harbouring developing and infective larvae were observed during the dry season in October and December when the monthly transmission potential recorded was 31 larvae/man/month. This dropped to 28 larvae/man/month and 25 larvae/man/month in January and February respectively. An annual transmission potential of 115 larvae/man/year was recorded., Conclusion: In spite of the ivermectin treatment in the area at the time of study, the transmission of Onchocerca volvulus continued unabated in the area with a higher transmission in the dry season than the rainy season. The distribution of ivermectin should be intensified through improved coverage while other control measures may be necessary during the dry season which is the peak of transmission in the area.

To assess the role of Aedes albopictus in transmission of filarial nematodes of veterinary importance, researches were carried out in different geographical areas. In Italy a first research was performed to study the susceptibility of Ae. albopictus to Dirofilaria repens, D. immitis and Setaria labiatopapillosa. The development of L3 larvae was longer than in other species of mosquitoes but Ae. albopictus could be a suitable vector of filariae. To understand the role of Ae. albopictus in the natural transmission of Dirofilaria and to assess the risk for animal and human health, in 2000, 2001 and 2002 another study was carried out in the town of Padua. A total of 2,534 Ae. albopictus were caught on human-attracted mosquitoes. Specific primers and sequencing identified filarial DNA as D. immitis; Ae. albopictus was proved a natural vector of D. immitis. Similar results were confirmed in Central Italy also for D. repens. The presence of Ae. albopictus increased the probability of transmission of canine and human dirofilariosis in urban environment and it could change the epidemiology of dirofilariosis, in particular for what concern the time of biting and the risk season. These aspects must be considered to outline a correct prophylaxis.

The text summarizes the principal current fields of investigation and the recent achievements of the research groups presently contributing to the Molecular Entomology Cluster of the Italian Malaria Network. Particular emphasis is given to the researches with a more direct impact on the fight against malaria vectors.

Seasonal and daily biting activity patterns, and natural filarial infections of adult black flies attracted to human bait were investigated at Ban Pang Faen, a rural area in Chiang Mai Province in northern Thailand. Collections were carried out twice a month from 0600 to 18-00 hours from January 2005 to February 2006. Among ten Simulium species collected, S. nodosum and S. asakoae were predominant occupying 57.3% and 37.2% of the total 16,553 females, respectively. These two predominant species showed different patterns in seasonal abundance: majority of S. nodosum (86.7%) were collected in hot season (from mid February to mid May), while most of S. asakoae (74.5%) were collected in rainy season (from mid May to mid October). For the daily biting activity, S. nodosum had two patterns: the main one was unimodal with a peak from 17-00 to 18-00, and the other was bimodal and had the major peak from 1600 to 18-00 and the minor one from 07-00 to 09-00. The pattern of S. asakoae was mostly unimodal with a peak from 06-00 to 10-00. The filarial larvae found in S. nodosum and S. asakoae were morphologically different from each other. The short and thick infective larvae found in S. asakoae differed from all known filarial larvae; it is suggested that they might be a bird parasite, Splendidofilariinae or Lemdaninae. The infection of the mammophilic S. nodosum with large Onchocerca type infective larvae was confirmed in this area. Natural filarial infections were found in each month (except December) in either S. nodosum or S. asakoae or in both. Monthly infection rates with all stages of larvae were 0.6-5.0% for S. nodosum, and 1.0-4.0% for S. asakoae. It is suggested that people in this village are exposed to the risk of infection with zoonotic filariae throughout the year.

Textile materials treated with synthetic repellents have the potential to provide protection against insect disease vectors but lack the residual activity necessary to achieve a prolonged effect or to be cost-effective. DEET MC is a formulation of DEET (N,N diethyl-m-toluamide) in which the repellent is gradually released from a capsule that binds the repellent. An experiment carried out on DEET-treated mosquito netting showed that the formulation repels, inhibits blood-feeding and kills mosquitoes for a period of at least 6 months under laboratory conditions. Such formulations may have the potential for use on nets against pyrethroid-resistant mosquitoes or on clothing or bedding materials distributed in disasters, emergencies or refugee camp situations.

The impact of climate on the vector behaviour of the worldwide dog tick Rhipicephalus sanguineus is a cause of concern. This tick is a vector for life-threatening organisms including Rickettsia rickettsii, the agent of Rocky Mountain spotted fever, R. conorii, the agent of Mediterranean spotted fever, and the ubiquitous emerging pathogen R. massiliae. A focus of spotted fever was investigated in France in May 2007. Blood and tissue samples from two patients were tested. An entomological survey was organised with the study of climatic conditions. An experimental model was designed to test the affinity of Rh. sanguineus for biting humans in variable temperature conditions. Serological and/or molecular tools confirmed that one patient was infected by R. conorii, whereas the other was infected by R. massiliae. Dense populations of Rh. sanguineus were found. They were infected with new genotypes of clonal populations of either R. conorii (24/133; 18%) or R. massiliae (13/133; 10%). April 2007 was the warmest since 1950, with summer-like temperatures. We show herein that the human affinity of Rh. sanguineus was increased in warmer temperatures. In addition to the originality of theses cases (ophthalmic involvements, the second reported case of R. massiliae infection), we provide evidence that this cluster of cases was related to a warming-mediated increase in the aggressiveness of Rh. sanguineus, leading to increased human attacks. From a global perspective, we predict that as a result of globalisation and warming, more pathogens transmitted by the brown dog tick may emerge in the future.

We have investigated the pathogenicity of tsetse (Glossina pallidipes)-transmitted cloned strains of Trypanosoma brucei rhodesiense in vervet monkeys. Tsetse flies were confirmed to have mature trypanosome infections by xenodiagnosis, after which nine monkeys were infected via the bite of a single infected fly. Chancres developed in five of the nine (55.6%) monkeys within 4 to 8 days post infection (dpi). All nine individuals were successfully infected, with a median pre-patent period of 4 (range = 4-10) days, indicating that trypanosomes migrated from the site of fly bite to the systemic circulation rapidly and independently of the development of the chancre. The time lag to detection of parasites in cerebrospinal fluid (CSF) was a median 16 (range = 8-40) days, marking the onset of central nervous system (CNS, late) stage disease. Subsequently, CSF white cell numbers increased above the pre-infection median count of 2 (range = 0-9) cells/microl, with a positive linear association between their numbers and that of CSF trypanosomes. Haematological changes showed that the monkeys experienced an early microcytic-hypochromic anaemia and severe progressive thrombocytopaenia. Despite a 3-fold increase in granulocyte numbers by 4 dpi, leucopaenia occurred early (8 dpi) in the monkey infection, determined mainly by reductions in lymphocyte numbers. Terminally, leucocytosis was observed in three of nine (33%) individuals. The duration of infection was a median of 68 (range = 22-120) days. Strain and individual differences were observed in the severity of the clinical and clinical pathology findings, with two strains (KETRI 3741 and 3801) producing a more acute disease than the other two (KETRI 3804 and 3928). The study shows that the fly-transmitted model accurately mimics the human disease and is therefore a suitable gateway to understanding human African trypanosomiasis (HAT; sleeping sickness).

A study was carried out to identify the native and exotic mosquito species that feed on possums (and also humans) during daytime in a native forest fragment in the Auckland region. Twenty-two possums were handled in spring 2005, and 21 in the following summer. A total of 32 female mosquitoes were collected while handling the possums (22 mosquitoes were on humans--all introduced Aedes notoscriptus; 10 on possums--9 Ae. notoscriptus and 1 native Coquillettidia iracunda). These results support previous findings that Aedes notoscriptus may regularly take blood meals from brushtail possums in New Zealand, as happens in Australia. Significant is the record of Coquillettidia iracunda feeding on a possum, which seems to be the first record of a native mosquito feeding on these animals in New Zealand. The potential public health implications of these findings are discussed.

Infection by Plasmodium, the causative agent of malaria, starts when the parasite, injected by a mosquito vector, reaches and invades the liver, where it transforms into a stage that is capable of infecting erythrocytes and that causes the symptoms and complications of the disease. This phase of the infection, called pre-erythrocytic stage, is the most elusive of the parasite's life cycle, yet it was identified more than fifty years ago as a primary target of vaccine strategies aimed at avoiding erythrocyte infection. Recently in vivo imaging in a rodent model revealed that the pre-erythrocytic phase is unexpectedly complex. In particular, it includes a component of lymphatic infection, thus altering our representation of how an immune response can be mounted against these parasite stages.

Mansonella perstans filariasis is widely present in Africa and equatorial America and its pathogenicity has been recently reconsidered. Although M. perstans infection has been considered a minor filariasis, remaining asymptomatic in most of infected subjects, more recent studies have shown that M. perstans is capable of inducing a variety of clinical features, including angioedemas, swellings like the "Calabar swellings" of loiasis, pruritus, fever, headache, pain in bursae and/or joint synovia, or in serous cavities. It is likely that some of the pathological changes observed are induced by the immune response to the infection. Eosinophilia is present in many cases of infection. Moreover M. perstans filariasis is difficult to be treated. Effective treatment is lacking and there is no consensus on optimal therapeutic approach. The most commonly used drug is diethylcarbamazine (DEC) that is however often ineffective. Although other drugs have been tried (e.g. praziquantel, ivermectin), none has proven to be reliably and rapidly effective. Mebendazole seemed more active than DEC in eliminating the infection, with a comparable rate of overall responses. Thiabendazole evidenced a small, but significant activity against the infection. Combination treatments (DEC plus mebendazole) resulted in a significantly higher activity compared with the single drugs.

Autochthonous transmission of the Chagas disease parasite, Trypanosoma cruzi, was detected in a patient in rural New Orleans, Louisiana. The patient had positive test results from 2 serologic tests and hemoculture. Fifty-six percent of 18 Triatoma sanguisuga collected from the house of the patient were positive for T. cruzi by PCR.

Transmission of Plasmodium falciparum is hyperendemic in southern Zambia. However, no data on the entomologic aspects of malaria transmission have been published from Zambia in more than 25 years. We evaluated seasonal malaria transmission by Anopheles arabiensis and An. funestus s.s. and characterized the blood feeding behavior of An. arabiensis in two village areas. Transmission during the 2004-2005 rainy season was nearly zero because of widespread drought. During 2005-2006, the estimated entomologic inoculation rate values were 1.6 and 18.3 infective bites per person per transmission season in each of the two village areas, respectively. Finally, with a human blood index of 0.923, An. arabiensis was substantially more anthropophilic in our study area than comparable samples of indoor-resting An. arabiensis throughout Africa and was the primary vector responsible for transmission of P. falciparum.

Visceral leishmaniasis (VL) is a rare disease in Thailand. Only one previous case has been reported in which transmission was likely autochthonous. We conducted an investigation of a case of VL, which included serological and symptom surveys of people who lived near the case in Nan Province and Bangkok, serological surveys of domestic animals in his home village and sand fly surveys in his home village and in Bangkok. No humans interviewed met our case definition for possible VL. One hundred thirty-one villagers were seronegative for Leishmania antibodies. We found three cows and one cat that had positive direct agglutination tests for Leishmania spp, but we were unable to confirm current infection by PCR. Sand fly surveys showed that most of the flies were of the Sergentomyia genus, which has not previously been reported as a competent vector in Thailand. Nonetheless, we conclude, based on the patient's lack of travel outside Thailand and the presence of seropositive domestic animals in his home village, that he was most likely infected by the bite of a sand fly in Thailand. We believe this is the second case of autochthonously transmitted VL in Thailand.

Owing to the development of pyrethroid resistance in Anopheles gambiae, there is a need to develop chemical alternatives for use on mosquito nets. Synthetic insect repellents are widely used for personal protection as skin or clothing applications. The efficacy of repellent-treated nets (RTN) was evaluated in experimental huts in Côte d'Ivoire against pyrethroid-resistant populations of An. gambiae and Culex quinquefasciatus. The repellents tested were DEET (N,N-diethyl-3-methylbenzamide) at 7.9 g/m2 and two formulations of ethyl butylacetylaminopropionate (IR3535) at 7.6 g/m2 and 7.3 g/m2. Over 45 nights there was a 74-82% reduction in the number of An. gambiae entering the huts containing RTNs but no significant reduction in entry of C. quinquefasciatus. There was a 63-64% reduction in the proportion of An. gambiae blood feeding but no reduction in the proportion of C. quinquefasciatus blood feeding in huts with RTNs. An unexpected result was the 69-76% mortality of An. gambiae and 51-61% mortality of C. quinquefasciatus in huts containing RTNs. Treated filter paper bioassays in WHO test kits confirmed that confined contact with DEET induces mortality. The DEET-based product provided better and longer protection; tunnel test bioassays confirmed that residual activity lasted for up to 6 weeks. Application of repellents to nets warrants further investigation and development.

Malaria-infected mosquitoes feeding on a mammalian host inject sporozoites into the skin to induce a malaria infection. The numbers of sporozoites ultimately able to reach the liver may be important determinants of the characteristics of the ensuing blood infection. Because feeding mosquitoes not only inject sporozoites into the host but concomitantly ingest blood to obtain their bloodmeal, some sporozoites are re-ingested by the feeding mosquito. We studied transmission of fluorescent Plasmodium berghei sporozoites injected into mice by Anopheles stephensi mosquitoes and found that the numbers of sporozoites re-ingested by mosquitoes are comparable to numbers previously reported to be delivered directly into mice. Thus, re-ingestion of sporozoites likely plays a significant role in transmission dynamics of malaria by mosquitoes, and may account for the failure of some sporozoite-infected mosquitoes to induce a blood infection.

One of the most significant modern day efforts to prevent and control an arthropod-borne disease during a military deployment occurred when a team of U.S. military entomologists led efforts to characterize, prevent, and control leishmaniasis at Tallil Air Base (TAB), Iraq, during Operation Iraqi Freedom. Soon after arriving at TAB on 22 March 2003, military entomologists determined that 1) high numbers of sand flies were present at TAB, 2) individual soldiers were receiving many sand fly bites in a single night, and 3) Leishmania parasites were present in 1.5% of the female sand flies as determined using a real-time (fluorogenic) Leishmania-generic polymerase chain reaction assay. The rapid determination that leishmaniasis was a specific threat in this area allowed for the establishment of a comprehensive Leishmaniasis Control Program (LCP) over 5 mo before the first case of leishmaniasis was confirmed in a U.S. soldier deployed to Iraq. The LCP had four components: 1) risk assessment, 2) enhancement of use of personal protective measures by all personnel at TAB, 3) vector and reservoir control, and 4) education of military personnel about sand flies and leishmaniasis. The establishment of the LCP at TAB before the onset of any human disease conclusively demonstrated that entomologists can play a critical role during military deployments.

This article analyzes the results of a one-year entomological survey conducted near the Yalala rapids in the Inga region, Democratic Republic of Congo, prior to the implementation of Community-Directed Treatment with Ivermectin (CDTI). A vector control program had been undertaken from October 1969 to December 1980, 20 kilometers upstream from the study site. The results of the present study show that during 122 collection days, 5,824 females were captured, of which 3,899 were dissected; 2,001 (51.3%) were parous while 44 (2.2%) were infected while 4 (0.2%) were infective. The crude Annual Transmission Potential (ATP) was 65 infective larvae per man and per year at the river as against 12 in Yalala village located two kilometers from the rapids. The average biting rate was 5 flies/man/day during the dry season and 90 during the rainy season. The vector capacity of similium shows that the risk of transmission of Onchocerca volvulus to the human population was not only reduced in time (2-3 months), but was also negligible all year round.

Background: Leishmaniasis is a vector-borne disease in which Leishmania parasites are transmitted by the bite of phlebotomine sand flies. Amastigotes are ingested by the sand fly vector with a blood meal taken from an infected host. This is followed by their differentiation into metacyclic promastigotes which are selectively released and permitted to migrate interiorly so as to make them available for transmission by bite. However, the actual number of amastigotes ingested by the sand fly in the blood meal is not known., Objective: To investigate the minimum number of Leishmania major amastigotes required to cause an infection in Phlebotomus duboscqi following an infective blood meal., Design: A laboratory based study., Setting: Centre for Biotechnology Research and Development, Kenya Medical Research institute, Nairobi., Results: Dissection of all fed sand flies at six days post-infective blood meal revealed that blood containing one amastigote per 0.3 microl in a total volume of 0.5 ml was able to cause an infection in the sand flies, but very few sand flies got infected (7.6% and 9.6% respectively). Concentrations of ten amastigotes per 0.3 microl in 0.5 ml gave infection rates of 35.4% and 26.3% respectively, suggesting that even when the concentration of amastigotes in a bloodmeal was high, not all sand flies feeding on it were able to pick up the parasites., Conclusions: These observations suggests that one amastigote is sufficient to cause an infection to a sand fly and as a result of multiplication in the gut and the existence of mechanisms that increase the number of infective bites delivered by a female sand fly they are able to sustain the transmission of leishmaniasis in an area.

Historically, in vivo experiments of Plasmodium gallinaceum in chickens have caused high mortality. Perhaps because of this high mortality, it remains to be demonstrated whether recovered birds will resist a second episode of illness when re-exposed to infected mosquitoes. In the current study, groups of 10 chicks were infected with P. gallinaceum via mosquito bite. Parasitemia and anemia were followed by recovery in all birds, although they had persisting, low levels of parasitized erythrocytes (0.007 +/- 0.019%). Twenty-three days after the initial exposure, 10 recovered chicks were rechallenged with P. gallinaceum via mosquito bite; none of them developed clinical or hematological evidence of malaria, in contrast to matched control birds, which all became diseased (P < 0.001). Unlike previous studies, the current experiment had no mortality in chickens infected with P. gallinaceum by mosquito bite. Recovered birds resisted disease from re-exposure to the same organism. The duration and nature of immunity or premunition remain to be determined.

Malaria is a serious tropical disease. Joyce Skeet outlines how infection occurs and the advice that can be given to travellers to avoid contracting the condition. The need for chemoprophylaxis for those travelling to high-risk areas is also identified

Ngaoundere Gudali zebu cattle naturally exposed to Simulium damnosum s.l. and Culicoides spp. bites were examined during 4 years for O. ochengi adult worm acquisition, Onchocerca ochengi and Onchocerca gutturosa skin microfilaria dynamics, and IgG1 and IgG2 antibody subclass responses. Eleven animals acquired a total of 465 O. ochengi nodules (average of 17 per female and 72 per male). The O. ochengi nodule load was highly variable in individual animals and exacerbated in mature male cattle. Three patterns of acquisition of O. ochengi (resistant to new infestation, early susceptibility and late susceptibility), not associated with Simulium biting intensity (P > 0.05), were distinguished. The minimum prepatent periods for O. ochengi nodules, O. ochengi microfilariae and O. gutturosa microfilariae were 10, 20 and 21 months, respectively. The O. ochengi microfilaria density significantly (P < 0.001) increased with age, was higher in young mature bulls than female animals (P < 0.001) and finally reached highest levels (P < 0.005) during the dry season. Antibody responses to Ov10/Ov11 recombinant O. volvulus antigens were predominantly of the IgG1 subclass. High levels of this subclass (not IgG2) observed in new born calves declined to almost zero levels at the age of 5-8 months but IgG1 levels significantly increased (P < 0.05) with age subsequently during patency. Put together the acquisition and accumulation of O. ochengi parasites in zebu cattle, apart from being season, sex (gender) and host age associated, may also suggest a density-dependent regulation of parasite establishment in a proportion of the exposed population.

Objective: To determine the biting activity and onchocerciasis transmission in Kashoya-Kitomi focus, Western Uganda., Design: Cross-sectional survey in randomly selected sites., Setting: Three districts in Western Uganda., Method: Crab trapping and examination for immature stages of Simulium neavei and full day human bait catches and dissections were conducted at three sites., Results: A total of 338 crabs were caught and out of this 372 (95.9%) were positive with immature stages of S. neavei with mean S. neavei/crab of 6.9. High biting density of S. neavei was observed in the two sites in Kabarole and Mbarara districts whereas an extremely low density was observed at a site in Bushenyi. Diurnal biting pattern revealed two peaks in the morning (09:00-10:00 hours.) and in the late afternoon (14:00-15:00 hours.) with a mid-day lull. The infection/infective rates were 53.3/11.0%, 30.7% to 7.8% and 20.0/0% for sites in Mbarara, Kabarole and Bushenyi districts, respectively. Monthly biting rates/ Transmission potentials for Mbarara, Kabarole and Bushenyi districts were 6231/5437, 9244/2916 and 247/0., Conclusion: There was high transmission of onchocerciasis observed in Kashoya-Kitomi focus despite the ongoing ivermectin treatment. The need for improved chemotherapeutic coverage and instituting other control measures may be necessary.

Despite the relatively high frequency of imported malaria in metropolitan France, the transmission of malaria by transfusion is exceptional. The screening of donations to determine those at risk is performed by an interview, and by the testing of serology for defined groups of donors. However, the exclusion of a candidate 'at risk' as a blood donor, by a pre-donation interview, is not completely mastered and the discrimination by biological examination lacks sensitivity, as much for methodological reasons as for reasons linked to the complex parasitic pathogenic agent (Plasmodium ssp.), as for the specific host defence system. The risk of introducing an unsafe-potentially dangerous (transfusion-transmitted malaria is often lethal)-element into the transfusional circuit is not completely covered. Is serology testing the most adequate test to avoid the risk of infected donations, in particular by Plasmodium falciparum; what are the alternatives and what will be the eventual added-costs of the biological qualification of such donations? The transfusional risk linked to Plasmodium seems, however, to be reduced to a minimum, concerning the circulation of plasma, which could represent an alternative for donors at real risk (rare) and those with a supposed risk (relatively numerous).

Objective: To study the risk of Trypanosoma cruzi domestic transmission using an entomological index and to explore its relationship with household's characteristics and cultural aspects., Methods: There were studied 158 households in an endemic area in Argentina. Each household was classified according to an entomological risk indicator (number of risky bites/human). A questionnaire was administered to evaluate risk factors among householders., Results: Infested households showed a wide range of risk values (0 to 5 risky bites/human) with skewed distribution, a high frequency of lower values and few very high risk households. Of all collected Triatoma infestans, 44% had had human blood meals whereas 27% had had dogs or chickens blood meals. Having dogs and birds sharing room with humans increased the risk values. Tidy clean households had contributed significantly to lower risk values as a result of low vector density. The infested households showed a 24.3% correlation between time after insecticide application and the number of vectors. But there was no correlation between the time after insecticide application and T. infestans' infectivity. The statistical analysis showed a high correlation between current values of the entomological risk indicator and Trypanosoma cruzi seroprevalence in children., Conclusions: The risk of T. cruzi domestic transmission assessed using an entomological index show a correlation with children seroprevalence for Chagas' disease and householders' habits.

An experiment was carried out in Burkina Faso to evaluate the potential for mechanical transmission of Trypanosoma vivax by the African tabanid Atylotus fuscipes. The experiment was carried out in a corral (10 m x 10 m) completely covered by a mosquito net (12 m x 12 m and 2.5m high). Eight heifers (cross-bred Zebu X Baoulé), free of trypanosome infection, were kept together with two heifers experimentally infected with a local stock of T. vivax. An average of 539 A. fuscipes per day, freshly captured with two Nzi traps, were introduced into the mosquito net from Day 1 to 20, to allow mechanical transmission of the parasites among cattle. Daily parasitological examinations (BCM) of cattle blood samples indicated that six of the eight receiver heifers were positive from days 9, 10, 15, 16, 19 and 29. Mechanical transmission of T. vivax by A. fuscipes was demonstrated unequivocally in close to natural conditions, at a high rate (75% incidence over a 20-day period) under a mean challenge of 54 insects per heifer per day. These results, in addition to previous demonstration of mechanical transmission of T. vivax by Atylotus agrestis, confirm that mechanical transmission can be a significant route of infection.

Objective: To describe an outbreak of Plasmodium vivax malaria in Far North Queensland in 2002., Design: Epidemiological and entomological investigations; molecular analyses of the infecting parasites., Main Outcome Measures: Case characteristics, adult and larval mosquito counts at the outbreak location, haplotyping of parasites in blood samples from different cases determined through sequencing of AMA1 and MSP1 genes., Results: A man with imported P. vivax malaria stayed at a camping ground 95 km north of Cairns in late September 2002. This led to an outbreak of P. vivax malaria in 10 adults who stayed at the camping ground in October. Large numbers of Anopheles farauti sensu lato larvae were present in stagnant pools in a creek at the camping ground, and many adult mosquitoes were collected nearby. Not only had most of the infected patients been exposed to mosquitoes at night, they were also less likely than other campers to have used insect repellents appropriately (odds ratio, 0.01; P < 0.001). Two different haplotypes of P. vivax, only one of which was detected in the imported case, were involved in the outbreak., Conclusions: Although local transmission of malaria is rare in Far North Queensland, the risk is probably higher in the dry season (September to December). Campers need to be aware of the increased risk of mosquito-borne diseases. Sexual recombination of multiple gametocytes in mosquitoes infected by the imported case may have resulted in the two haplotypes of P. vivax involved in the outbreak.

Investigations were carried out in Padova town (Veneto region, NE Italy) to define the actual role of Aedes albopictus in the natural transmission of Dirofilaria nematodes, and to assess the risk that its presence might represent for veterinary and medical health. During summer 2000-2002 daytime captures of human-attracted mosquitoes were carried out in three areas of the town. The presence of filarial parasites in mosquitoes was evaluated by PCR, and sequencing confirmed species assessment. DNA extraction was performed separately on pools of the insect abdomen and thorax-head, to discriminate between Dirofilaria infected/infective specimens. A total of 2721 mosquitoes were caught and A. albopictus was the most abundant species (2534). Filarial DNA was found in 27.5% (19/69) of the abdomen pools formed with mosquitoes collected in summer 2000, and in 11.1% (16/144) and 4.9% (6/123) thorax-head pools coming from samplings 2001 and 2002, respectively. Filarial DNA was belonging to D. immitis and all studied areas harboured infective specimens. These results prove A. albopictus as natural vector of D. immitis in Italy. Moreover, they support the hypothesis that the presence of the mosquito could affect the transmission pattern of canine heartworm disease in urban environment and, considering the aggressive anthropophylic behaviour of the species (30-48 bites/h) proven in Padova town, could enhance the circulation of filarial nematodes from animals to humans.

Phlebotomus sergenti is a natural vector of Leishmania tropica. However, the ability of P. sergenti to transmit L. tropica by bite has not been proven experimentally yet. We have transmitted L. tropica to golden hamsters and BALB/c mice by the bite of P. sergenti. Sand flies and Leishmania both originated from an anthroponotic cutaneous leishmaniasis focus in Urfa, Turkey. P. sergenti females from a laboratory colony were infected by feeding on lesions of needle-inoculated hamsters or mice. Gravid females were allowed to refeed on uninfected hosts 9-15 d after the infective feeding. At the second feeding, some infected females took a full blood meal, while others only a partial one; some females failed to feed at all. The ability of infected females to take a blood meal did not correlate with the parasite transmissibility. In four BALB/c mice, lesions developed after 1-6 months. In two albino hamsters (Mesocricetus auratus), lesions developed 1 month after the infective feeding, and Leishmania could be reisolated from these sites. Another hamster did not develop a lesion; however, the feeding site and the adjacent ear were PCR positive 1 year after infective feeding. Our results show that dissemination to other parts of host body occurs in L. tropica after sand fly bite. Experimental transmission of the parasite confirms that P. sergenti is a natural vector of L. tropica.

Two forms of leishmaniasis are endemic to the Jenin district in the northern region of the West Bank. Visceral leishmaniasis (VL), caused by Leishmania infantum, mainly affects infants. Cutaneous leishmaniasis (CL) affects a broader age group and is probably caused by L. tropica. Although the Jenin district is the most important focus of leishmaniasis in the West Bank, the sand fly fauna of the area has never been studied in a systematic manner. We collected base-line data on sand fly species, their distribution, and their feeding preferences to facilitate risk assessments for contracting leishmaniasis. Light traps, sticky traps, insecticide knockdown collections, aspirator, and human-landing collections were used. A total of 4,082 sand flies was collected in foci of confidence limits and/or VL between June and December 1998. Nine Phlebotomus species representing seven subgenera were identified: P. (Larroussius) perfiliewi transcaucasicus Perfil'ev, P. (La.) tobbi Adler & Theodor, P. (La.) mascitti canaaniticus Adler & Theodor, P. (La.) mascitti mascitti Grassi, P. (La.) syriacus Adler & Theodor, P. (Phlebotomus) papatasi Scopoli, P. (Synphlebotomus) s.p., P. (Paraphlebotomus) sergenti Parrot, P. (Par.) jacusieli Theodor, P. (Adlerius) halepensis Theodor. Two other Phlebotomus subspecies, P. (La.) major major Annandale, P. (La .) neglectus Tonnoir, require confirmation. In addition, four species of the closely related genus, Sergentomyia were also found: S. (Sergentomyia) theodori Parrot, S. (S.) fallax Parrot, S. (Sintonitus) tiberiadis Adler, Theodor & Lourie, S. (Sin.) christophersi Sinton. Among five species of sand flies collected on human bait, P. papatasi constituted approximately 90% followed by P. major syriacus (8%) and P. mascitti (2%). Sand fly human-biting activity occurred through the night and it was highest between 2400 and 0300 hours. P. papatasi. P. perfiliewi, P. major and P. tobbi were the more endophilic species constituting 93% of all flies caught indoors. Seven Phlebotomus spp. constitute potential vectors of leishmaniasis but the most probable ones are as follows: P. papatasi the main human-biting species, a recognized vector of L. major (CL), P. sergenti, L. tropica (CL) and P. syriacus, L. infantum (VL).

To assess the relationships between variations of Plasmodium falciparum transmission and those of peripheral parasitaemia prevalence or malaria attack incidence rates in regions with limited fluctuations of transmission, we conducted a follow-up in two Gabonese populations. Entomological surveys were carried out from May 1995 to April 1996 in Dienga, and from May 1998 to April 1999 in Benguia. In Dienga, malaria transmission was seasonal, being not detected during two 3-month periods. Mean entomological inoculation rate (EIR) was 0.28 infective bite/person/night. In Benguia, malaria transmission was perennial with seasonal fluctuations, mean EIR being 0.76 infective bite/person/night. In Dienga, 301 schoolchildren were followed from October 1995 to March 1996. Clinical malaria attack was defined as fever associated with >5000 parasites/microl of blood. P. falciparum prevalence varied from 28 to 42%, and monthly malaria attack incidence from 30 to 169 per thousand. In Benguia, the entire population (122 persons) was followed from November 1998 to April 1999. Prevalence varied from 22 to 50%, and monthly malaria attack incidence from 52 to 179 per thousand. In each area, entomological variations were not related to parasite prevalence, but preceded malaria attack incidence with 1- or 2-month time lag, corresponding to the pre-patency period that differs in the two populations, possibly according to differences in immunity related to parasite transmission.

The role of mechanical vectors in the transmission of African livestock trypanosomes has always been controversial relative to tsetse flies, their cyclical vectors. An experiment was carried out in Burkina Faso to demonstrate mechanical transmission of Trypanosoma vivax by one of the most common tabanids in Africa: Atylotus agrestis. Eight heifers (crossbred zebuxBaoulé), free of trypanosome infection, were kept in a corral covered by a mosquito net, together with two heifers infected experimentally with a local stock of T. vivax. On average, 324 A. agrestis, freshly captured with Nzi traps, were introduced daily over 20 days. Parasitological, PCR and serological examinations were carried out regularly to assess infections and levels of parasitaemia. Microscopic examination of buffy-coats indicated that five of the eight receiver-heifers were infected on days 8, 13, 32, 41, and 48. PCR results indicated that these five heifers were already infected by day 13. Mechanical transmission of T. vivax by A. agrestis was demonstrated unequivocally, at a high rate (63% in 13-20 days). Conditions of transmission in this experiment are discussed in terms of natural rates of challenge. The importance of tabanids as mechanical vectors of T. vivax should be re-considered, in light of these results. Creation of tsetse free zones in Africa will generally lead to the disappearance of T. congolense, T. brucei, and most often T. vivax as well; however, in areas where T. vivax can be mechanically transmitted, clearance of tsetse may not be sufficient to eradicate livestock trypanosomosis.

Anophelines were sampled from 82 locations on Buka and Bougainville islands in Papua New Guinea by larval collections, carbon dioxide-baited mosquito traps, and human biting catches. Anopheles farauti s.s. was collected in larval surveys but infrequently in mosquito traps on both islands; on Buka Island this species was readily collected in human biting catches. Anopheles farauti 2 was commonly collected in larval surveys on both islands; however, it was not collected in either mosquito traps or human biting catches. Anopheles punctulatus was found only on Buka Island, where it was commonly collected as larvae, but rarely in human biting catches and mosquito traps. Anopheles lungae was collected as larvae from only 1 site on Bougainville. Anopheles farauti s.s. fed consistently throughout the night (1900-0600 h); small peaks at midnight and dawn were not statistically significant. Of 1,156 An. farauti s.s. specimens examined by enzyme-linked immunosorbent assay for malaria sporozoites, 20 were found to be positive; 12 were positive for Plasmodium falciparum and 8 were positive for P. vivax (247 variant = 5; 210 variant = 3). Anopheles farauti s.s. seems to be the major malaria vector on these islands, whereas An. punctulatus may play a minor role on Buka Island. Anophele jarauti 2 is unlikely to be involved in malaria transmission on Buka or Bougainville islands.

An epidemiological study on malaria was undertaken between July 1995 and July 1996 in two villages (Zaïpobly and Gahably) and their encampments (Kouassikro, Hamanikro and Konankro), in the south-western forest area of Côte d'Ivoire (region of Taï). The parasitological scheme comprised a total of 2023 tests performed on children aged from 0 to 14 years. The species found were Plasmodium falciparum, P. malariae and P. ovale with a proportion of 84%, 14% and 2% respectively. The global parasite prevalence of all Plasmodium species was 85% and malaria was holoendemic. The average parasitic density decreased progressively as the age increased, in contrast to the plasmodic index which did not vary. All the malarial indexes were similar in the villages and their encampments. Only overall fever prevalence was permanent and in all age groups it was higher in the encampments than in the villages. The entomological findings showed that transmission was permanent and intense throughout the year, with a recrudescence during the rainy season. Transmission was attributed to Anopheles gambiae s.l. in 85% of the cases whereas An. funestus played a secondary role. The average sporozoïtic index was 7.6% and varied between 1.1% and 16.7%. The entomological inoculation rate was of 400 infected bites per person-year for An. gambiae s.l. In such conditions of intense transmission, acquisition of premunition starts at a very early age. This assertion is verified by the average parasite density and the frequency of high parasitic densities which were at their maximum between 1 and 4 years of age and decreased thereafter as the age increased. The paludometric and entomologic indexes obtained are the most elevated ever to have been observed in Côte d'Ivoire, as a result of considerable ecological changes linked to the deterioration of the forest environment over the past 30 years. This deterioration has probably been caused by demographic pressure resulting from internal and foreign immigration to the Taï region and more especially by the influx of Liberian refugees.

In the Zambezi valley, mosquito females of the Anopheles gambiae Giles complex (Diptera: Culicidae) were collected from a hut containing pairs of cattle distinguishable by known DNA markers. DNA was extracted from the blood-fed mosquito abdomens and primer sets for ungulate and mosquito DNA loci were used to identify the mosquito sibling species and individual host source(s) of their bloodmeals. The 67 mosquitoes comprised a mixture of An. arabiensis Patton (31%) and An. quadriannulatus Theobald (69%). DNA from one or both of the cattle present in the hut was detected in 91% of samples. When the hut contained an adult and a calf, the percentage of bloodmeals from the adult, the calf and adult + calf were 58%, 27% and 15%, respectively; the trend towards meals from the adult host was consistent but not always significant. When the pair of cattle comprised two adults of roughly equal size and age, then mosquitoes generally showed no significant bias towards feeding from one individual. There was no significant difference in the pattern of host selection made by An. arabiensis and An. quadriannulatus but the former had a significantly higher percentage (20%) of mixed meals than An. quadriannulatus (9%). These two members of the An. gambiae complex appear to be less selective in their choice of cattle hosts compared to day-active Diptera such as tsetse and Stomoxys, possibly because the hosts are generally asleep when Anopheles are active and there is therefore less selective pressure to adapt to host defensive behaviour. The slight bias of Anopheles towards older and/or larger cattle may be related to the host's larger surface area.

In Zimbabwe, observations were made of the behaviour of individual stable flies (Stomoxys spp.) (Diptera: Muscidae) and tsetse (Glossina spp.) (Diptera: Glossinidae) feeding on cattle during the wet (Stomoxys and tsetse) and dry (tsetse only) seasons. For Stomoxys landing on adult cattle, only 27% took a full meal (mean feeding time = 147 s). Most Stomoxys left the host before completing their meal, largely due to disturbance by the host's defensive behaviour (24%, mean time = 59 s) or other flies (44%, 71 s). The probability of a Stomoxys leaving the host progressively increased with time. Simultaneous observations of tsetse showed that, compared to Stomoxys, their feeding success was lower (15%), feeding was interrupted earlier (33 s) and the time taken to complete a meal was shorter (109 s). Further studies of tsetse across different seasons and hosts showed that feeding success varied according to host age (adult = 7%; calf = 3%) and was negatively correlated with the frequency of host defensive behaviour and the relative abundance of non-biting Diptera. Disturbances were more often caused by host behaviour (69%) than other flies (31%) and the probability of tsetse leaving decreased with time on the host. Overall, these results suggest that tsetse and Stomoxys have different feeding strategies. In particular, tsetse appear to be more responsive to host defensive behaviour, which reduces their feeding success relative to Stomoxys. These behavioural differences are consistent with the respective life-history characteristics of Stomoxys and tsetse.

We studied the biting densities of Chrysops silacea and the transmission of loiasis over 1 year in a regenerated forest in the south-west province of Cameroon. A total of 3015 flies caught near a wood fire at ground level during rainy and dry seasons were identified morphologically and 1975 caught during the rainy season were dissected to determine their physiological age and infection rate. The prevalence of microfilaraemia in the human population in the study area was determined using the thick blood smear method. Chrysops silacea was the only species caught. The daily and seasonal biting cycle of C. silacea showed two peaks of activities, 9-11 a.m. and 2-4 p.m. The biting cycles of parous and nulliparous flies showed the same trends, but the density of nulliparous flies biting at all time of the day was 2-3 times higher. Chrysops silacea biting density was high during the rainy season (9.06 +/- 6.88 flies/man/h) and lowest during the dry season (0.44 +/- 0.75 flies/man/h). An infection rate of 1.72% and a monthly morning and afternoon transmission potentials of 120769.11 and 139016.64 infective head L3/man were observed, respectively, in the rainy season. Even though few Chrysops carried Loa loa infective larvae (0.7%), their parasite load was high, giving a high level of transmission of L. loa in the area. A total of 20.37% of the people examined for blood microfilariae were positive. These results suggest that the study area is an active focus of loiasis transmission.

The requirements for maintenance of antigen (Ag)-specific memory T cells in protection to malaria is poorly understood. We have previously demonstrated a recall of IL-4-producing memory CD4(+)CD45RO(+) T cells with parasitized red blood cells (pRBC) in persons protected by radiation-attenuated Plasmodium falciparum sporozoites (gamma-spz). Using the CD27 marker, we have now identified two subsets of CD4(+)CD45RO(+) T cells: CD4(+)CD45RO(+)CD27(+) T cells representing an early memory and CD4(+)CD45RO(+)CD27() T cells representing a terminally differentiated memory cells. A small subset of CD4(+)CD45RO(+)CD27(-) T cells also expressed CD70, the CD27 ligand. The addition of anti-CD70 monoclonal antibody (mAb) to pRBC-stimulated cultures significantly inhibited the conversion of CD27(+) to CD27(-) subset without profoundly affecting IL-4 production. In contrast, the inclusion of anti-CD27 mAb in parallel cultures abrogated IL-4 production without interfering with conscription of T cells into the CD27(-) T cell set. We propose that the persistence of memory CD4(+) T cells depends on Ag-driven conscription of a mature memory phenotype through co-ligation of CD27 and CD70 expressed, respectively, on CD27(+) and CD27(-) T cells. Hence, protracted protection in malaria depends in part on memory CD4(+) T cells that require specific Ag presumably from the repositories of liver-and blood-stage antigens and the delivery of a second signal from the CD27:CD70 interaction.

Canine visceral leishmaniasis (VL), caused by Leishmania infantum (Leishmania chagasi in the New World), is a zoonotic, endemic disease in Western Europe and Latin America. The potential spreading to new regions was suggested by the appearance of canine VL among foxhounds in the US. Although the sand fly vectors in the major foci of transmission have been described, no information exists on other sand flies that could propagate the infection outside endemic areas. We evaluated the capacity of Lutzomyia shannoni (Dyar) and Lutomyia youngi (Feliciangeli & Murillo), which are widely distributed in the New World, to acquire L chagasi (Cunha and Chagas) infections. A high proportion of L youngi were infected after feeding on an oligosymptomatic dog (51 per cent) or a polysymptomatic individual (95 per cent), but the intensity of infection was low (< 200 promastigotes/fly). L shannoni became infected only by feeding on the polysymptomatic dog, and the infection rate was lower (9 per cent) than in Lutzomyia longipalpis (36 per cent), and Lutzomyia evansi (Nunez-Tovar) (Lutz and Neiva) (38 per cent), but the intensity of infection (200 to > 500 promastigotes/fly) was comparable (L longipalpis) or higher (L evansi) than in the New World vectors. It is hypothesised that the presence of infected dogs in areas where L shannoni or L youngi occur could initiate new endemic cycles of VL in both South and North America.

Much progress has been made in describing how it is, in a mechanistic sense, that some vertebrate hosts (species or individuals) are bitten more than others, principally because of their odour or appearance. Little attention has been paid to why, in an evolutionary sense, these particular animals are bitten. Irrespective of the proximate mechanisms of host choice, there must be an intense selection pressure on insects to feed on those hosts that are most amenable to being bitten. We should be better able to predict host choice by understanding the evolutionary processes at work.

The probability of Trypanosoma cruzi transmission to opossums by independent events of predation and fecal contamination during feeding ("biting") with positive Triatoma infestans was estimated. Negative female opossums were challenged for 23 hr with 10 infected third and fourth instars of T. infestans, and tests for positivity for T. cruzi by xenodiagnosis were performed at 30, 60, and 90 days. From these data, seven probability parameters were estimated by maximum likelihood, and likelihood ratio statistics confidence intervals were calculated. Simultaneous estimation of p1 (probability that a "bite" will infect an opossum), p3 (probability that a bug that has been eaten by an opossum will infect it), and p6 (probability that the opossum will become infected if faced with an infected triatomine), resulted in p1 = 0.06, p3 = 0.075, and p6 = 0.059. On average, each opossum should be exposed to an average of 700 encounters with bugs during its life, resulting in about eight potentially infective contacts, to produce the 35% opossum prevalence found in the field.