Parasites and Vectors

Toscana virus isolated from sandflies, Morocco

To investigate the transmission of phleboviruses, a total of 7,057 sandflies were collected in well-known foci of cutaneous leishmaniasis and were identified to species level according to morphological characters.Collected sandflies were tested by Nested PCR for the presence of Phleboviruses and subsequently by viral isolation on Vero cells. The corresponding products were sequenced. Toscana virus was isolated, for the first time, from 5 pools of sandflies.Hence, Toscana virus should be considered a potential risk that threatens public health and clinicians should be aware of the role of Toscana virus in cases of meningitis and encephalitis in Morocco.

Efficacy of a fixed combination of permethrin 54.5% and fipronil 6.1% (Effitix®) in dogs experimentally infested with Ixodes ricinus

Background: Ticks are the most important vectors of disease-causing pathogens in domestic animals and are considered to be second worldwide to mosquitoes as vectors of human diseases. In Europe, Ixodes ricinus, the sheep tick, plays an important role as companion animal parasite but is also the primary vector of medically important diseases such as tick-borne encephalitis and Lyme borreliosis.The present study was designed to evaluate the efficacy under laboratory conditions of a new fixed spot-on combination of fipronil and permethrin (Effitix®, Virbac) in treating and preventing tick infestations of Ixodes ricinus in dogs. Methods: Twelve dogs were included in this randomized, controlled, blinded laboratory study. They were randomly allocated to two groups of six dogs each according to their pre-treatment live attached Ixodes ricinus tick count. On day 0, the dogs from Group 2 were treated with the recommended dose of Effitix®, the dogs from Group 1 remained untreated. On days −2, 7, 14, 21, 28 and 35, all dogs were infested with 50 (±4) viable unfed adult Ixodes ricinus (20 ± 2 males, 30 ± 2 females). Ticks were removed and counted at 48 ± 2 hours post product administration or tick infestations. Results: Through the study, the tick attachment rates for the untreated group were greater than 25% demonstrating that adequate levels of infestation were reached on the control dogs. Based on both arithmetic and geometric means (AM and GM), Effitix® was deemed to be effective against Ixodes ricinus on days 2, 9, 16, 23, 30 and 37 with a percentage of efficacy of 98%, 100%, 100%, 100%, 93% and 95% respectively (AM). No clinical abnormalities were detected during the study. Conclusions: The study has shown under laboratory conditions, that Effitix® is a safe and an effective combination to treat and protect dogs from Ixodes ricinus up to 37 days after administration. The high immediate efficacy of 98% evaluated at 48 hours post-treatment was particularly interesting, meaning that Effitix has a curative effect against ticks (Ixodes ricinus) and provides a rapid control of existing Ixodes ricinus infestation on a dog at the time of treatment.

The impact of different sprayable surfaces on the effectiveness of indoor residual spraying using a micro encapsulated formulation of lambda-cyhalothrin against Anopheles gambiae s.s.

Background: The type of sprayable surface impacts on residual efficacy of insecticide used in indoor residual spraying (IRS). However, there is limited data on common types of wall surfaces sprayed in Zanzibar and mainland Tanzania where IRS began in 2006 and 2007 respectively. The study investigated residual efficacy of micro-encapsulated lambda-cyhalothrin sprayed on common surfaces of human dwellings and domestic animal shelters in Zanzibar and mainland Tanzania. Methods: An experimental hut was constructed with different types of materials simulating common sprayable surfaces in Zanzibar and mainland Tanzania. Surfaces included cement plastered wall, mud-daub, white-wash, wood, palm-thatch, galvanized iron-sheets, burnt-bricks, limestone and oil-paint. The World Health Organization (WHO) procedure for IRS was used to spray lambda-cyhalothrin on surfaces at the dose of 20–25 mg/m2. Residual efficacy of insecticide was monitored through cone bioassay using laboratory-reared mosquitoes; Kisumu strain (R–70) of Anopheles gambiae ss. Cone bioassay was done every fortnight for a period of 152 days. The WHO Pesticide Evaluation Scheme (WHOPES) threshold (80% mortality) was used as cut-off point for acceptable residual efficacy. Results: A total of 5,800 mosquitos were subjected to contact cone bioassay to test residual efficacy of lambda-cyhalothrin. There was a statistically significant variation in residual efficacy between the different types of wall surfaces (r = 0.24; p < 0.001). Residual efficacy decreased with increasing pH of the substrate (r = −0.5; p < 0.001). Based on WHOPES standards, shorter residual efficacy (42-56days) was found in wall substrates made of cement, limestone, mud-daub, oil paint and white wash. Burnt bricks retained the residual efficacy up to 134 days while galvanized iron sheets, palm thatch and wood retained the recommended residual efficacy beyond 152 days. Conclusion: The study revealed a wide variation in residual efficacy of micro encapsulated formulation of lambda-cyhalothrin across the different types of wall surfaces studied. In areas where malaria transmission is bimodal and wall surfaces with short residual efficacy comprise > 20% of sprayable structures, two rounds of IRS using lambda-cyhalothrin should be considered. Further studies are required to investigate the impact of sprayable surfaces on residual efficacy of other insecticides commonly used for IRS in Zanzibar and mainland Tanzania.

Mitochondrial and nuclear ribosomal DNA dataset supports that Paramphistomum leydeni (Trematoda: Digenea) is a distinct rumen fluke species

Background: Rumen flukes parasitize the rumen and reticulum of ruminants, causing paramphistomiasis. Over the years, there has been considerable debate as to whether Paramphistomum leydeni and Paramphistomum cervi are the same or distant species. Methods: In the present study, the complete mitochondrial (mt) genome of P. leydeni was amplified using PCR-based sequencing and compared with that of P. cervi. The second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA) of P. leydeni specimens (n = 6) and P. cervi specimens (n = 8) was amplified and then sequenced. Phylogenetic relationship of the concatenated amino acid sequence data for 12 protein-coding genes of the two rumen flukes and selected members of Trematoda was evaluated using Bayesian inference (BI). Results: The complete mt genome of P. leydeni was 14,050 bp in size. Significant nucleotide difference between the P. leydeni mt genome and that of P. cervi (14.7%) was observed. For genetic divergence in ITS-2, sequence difference between P. leydeni and P. cervi was 3.1%, while no sequence variation was detected within each of them. Phylogenetic analysis indicated that P. leydeni and P. cervi are closely-related but distinct rumen flukes. Conclusions: Results of the present study support the proposal that P. leydeni and P. cervi represent two distinct valid species. The mt genome sequences of P. leydeni provide plentiful resources of mitochondrial markers, which can be combined with nuclear markers, for further comparative studies of the biology of P. leydeni and its congeners from China and other countries.

Dexamethasone inhibits brain apoptosis in mice with eosinophilic meningitis caused by Angiostrongylus cantonensis infection

Background: Angiostrongylus cantonensis, the rat lungworm, is the major cause of eosinophilic meningitis worldwide. Rats serve as the definitive host of the nematode, but humans can be infected incidentally, leading to eosinophilic meningitis. A previous BALB/c animal study has demonstrated increased apoptotic proteins and decreased anti-apoptotic proteins in mice infected with A. cantonensis. Steroids may be an effective treatment option for eosinophilic meningitis caused by A. cantonensis, but the involved mechanism is unclear. This study hypothesized that the beneficial effects of steroids on eosinophilic meningitis are mediated by decreased apoptosis. Methods: In a BALB/c animal model, mice were orally infected with 50 A. cantonensis L3 via an oro-gastric tube and were sacrificed every week for 3 consecutive weeks after infection or until the end of the study. Dexamethasone was injected intra-peritoneally from the 7th day post-infection until the end of the 21-day study. Evans blue method was used to measure changes in the blood brain barrier, while western blotting, immuno-histochemistry, and TUNEL assay were used to analyze brain homogenates expression of apoptotic and anti-apoptotic proteins. Results: There were increased amounts of Evans blue, apoptotic proteins (caspase-3, -8, and -9 and cytochrome C), and decreased anti-apoptotic proteins (bcl-2) after 2-3 weeks of infection. Dexamethasone administration significantly decreased Evans blue extravasations and apoptotic protein expressions. Conclusions: Apoptosis of mice brain homogenates can be repressed by dexamethasone treatment.

Situation analysis of parasitological and entomological indices of onchocerciasis transmission in three drainage basins of the rain forest of South West Cameroon after a decade of ivermectin treatment

Background: Community-Directed Treatment with Ivermectin (CDTI) is the main strategy adopted by the African Programme for Onchocerciasis control (APOC). Recent reports from onchocerciasis endemic areas of savannah zones have demonstrated the feasibility of disease elimination through CDTI. Such information is lacking in rain forest zones. In this study, we investigated the parasitological and entomological indices of onchocerciasis transmission in three drainage basins in the rain forest area of Cameroon after a decade of CDTI. River basins differed in terms of river number and their flow rates; and were characterized by high pre-control prevalence rates (60-98%). Methods: Nodule palpation and skin snipping were carried out in the study communities to determine the nodule rates, microfilarial prevalences and intensity. Simulium flies were caught at capture points and dissected to determine the biting, parous, infection and infective rates and the transmission potential. Results: The highest mean microfilaria (mf) prevalence was recorded in the Meme (52.7%), followed by Mungo (41.0%) and Manyu drainage basin (33.0%). The same trend was seen with nodule prevalence between the drainage basins. Twenty-three (23/39) communities (among which 13 in the Meme) still had mf prevalence above 40%. All the communities surveyed had community microfilarial loads (CMFL) below 10 mf/skin snip (ss). The infection was more intense in the Mungo and Meme. The intensity of infection was still high in younger individuals and children less than 10 years of age. Transmission potentials as high as 1211.7 infective larvae/person/month were found in some of the study communities. Entomological indices followed the same trend as the parasitological indices in the three river basins with the Meme having the highest values. Conclusion: When compared with pre-control data, results of the present study show that after over a decade of CDTI, the burden of onchocerciasis has reduced. However, transmission is still going on in this study site where loiasis and onchocerciasis are co-endemic and where ecological factors strongly favour the onchocerciasis transmission. The possible reasons for this persistent and differential transmission despite over a decade of control efforts using ivermectin are discussed.

Evolutionary history of Leishmania killicki (synonymous Leishmania tropica ) and taxonomic implications

Background: The taxonomic status of Leishmania (L.) killicki, a parasite that causes chronic cutaneous leishmaniasis, is not well defined yet. Indeed, some researchers suggested that this taxon could be included in the L. tropica complex, whereas others considered it as a distinct phylogenetic complex. To try to solve this taxonomic issue we carried out a detailed study on the evolutionary history of L. killicki relative to L. tropica. Methods: Thirty-five L. killicki and 25 L. tropica strains isolated from humans and originating from several countries were characterized using the MultiLocus Enzyme Electrophoresis (MLEE) and the MultiLocus Sequence Typing (MLST) approaches. Results: The results of the genetic and phylogenetic analyses strongly support the hypothesis that L. killicki belongs to the L. tropica complex. Our data suggest that L. killicki emerged from a single founder event and that it evolved independently from L. tropica. However, they do not validate the hypothesis that L. killicki is a distinct complex. Therefore, we suggest naming this taxon L. killicki (synonymous L. tropica) until further epidemiological and phylogenetic studies justify the L. killicki denomination. Conclusions: This study provides taxonomic and phylogenetic information on L. killicki and improves our knowledge on the evolutionary history of this taxon.

Knowledge, attitudes, and practices of Texas hunters: a potentially high-risk population for exposure to the parasite that causes Chagas disease

Background: Chagas disease is a parasitic infection transmitted to humans and mammals by the Triatominae insect. If untreated, Chagas disease can lead to heart failure and death. Previous publications highlighted the potential public health risk of disease transmission among hunters in the United States.FindingsWe further investigated this population’s risk by administering a knowledge, attitudes, and practices questionnaire. Responses from hunters detailed the vector exposure and hunting practices unique to this population that might lead to their increased risk of disease transmission. Conclusions: Hunters should be aware of their potential risk for exposure to the insect that could be infected with the parasite that causes Chagas disease.

Altered levels of circulating miRNAs are associated Schistosoma japonicum infection in mice

Background: Dioecious flatworms of the genus schistosoma causes schistosomiasis, which is a major public health problem in developing countries. Acquiring detailed knowledge of schistosome-host interactions may aid in the development of novel strategies for schistosomiasis control. MicroRNAs (miRNAs) are involved in processes such as development, cell proliferation, metabolism, and signal transduction. Circulating miRNAs not only serve as a novel class of biomarkers of many diseases but also regulate target gene expression in recipient cells, which are similar to hormones. Methods: In the present study, we used miRNA microarrays to determine the profile of circulating miRNAs associated with S. japonicum infection of mice. The biological functions of the altered levels of miRNAs and their target genes were predicted using bioinformatics. Expression levels of selected miRNAs and their target genes were further analyzed by quantitative RT-PCR. Results: Our study identified 294 and 189 miRNAs in infected mice that were expressed in two independent experiments at levels ≥ 2-fold higher or ≤ 0.5-fold lower, respectively, compared with uninfected mice. Thirty-six of the same miRNAs were detected in these analyses. Moreover, pathway analyses indicated that most of these miRNAs are putatively involved in signaling pathways associated with pathogenesis, such as Wnt and MAPK signaling. Further, we show an inverse correlation between the circulating levels of these miRNAs and their target genes, suggesting that changes in miRNA expression may cause aberrant expression of genes such as Creb1 and Caspase-3 in mice infected with S. japonicum. Conclusions: Our study shows significant differences in the levels of circulating miRNAs between S. japonicum infected mice and uninfected mice. In particular, the altered levels of miR-706 and miR-134-5p were associated with altered levels of expression of the Caspase-3 and Creb1 genes, respectively, suggesting that circulating miRNAs may serve as important mediators of the pathology of hepatic schistosomiasis. Additionally, our results are expected to provide new insights for further understanding the mechanisms of schistosome-host interaction that may facilitate in the development of novel interventions for alleviating the symptom of S. japonicum infection as well as for preventing and treating schistosomiasis.

Role of underappreciated vectors in malaria transmission in an endemic region of Bangladesh-India border

Background: Despite the efforts of the National Malaria Control Programme, malaria remains as an important public health problem in Bangladesh, particularly in the south-eastern region bordering India. Successful malaria control strategies rely on a detailed understanding of the underlying causes of malaria transmission. Here, an entomological survey was conducted in a malaria endemic area of Bangladesh bordering India to investigate the Anopheles mosquito community and assess their Plasmodium infection status. Methods: Monthly entomological collections were undertaken from October 2010 to September 2011 in five villages in the Matiranga sub-district, Khagrachari district in Bangladesh, bordering the Indian State of Tripura. CDC miniature light traps were placed inside houses to collect adult Anopheles mosquitoes. Following morphological and molecular identification of the female Anopheles mosquitoes collected, they were screened for circumsporozoite proteins (CSP) of Plasmodium falciparum (Pf), Plasmodium vivax-210 (Pv-210) and Plasmodium vivax-247 (Pv-247), by ELISA to determine natural infection rates. Variation in Anopheles species composition, relative abundance and Plasmodium infection rates were analysed between sampled villages. Results: A total of 2,027 female Anopheles were collected, belonging to 20 species. Anopheles nivipes was the most abundant species in our test villages during the peak malaria transmission season, and was observed sympatrically with An. philippinensis in the studied area. However, in the dry off-peak season, An. jeyporiensis was the most abundant species. Shannon’s diversity index was highest in October (2.12) and evenness was highest in May (0.91). The CSP ELISA positive rate overall was 0.44%. Anopheles karwari (n = 2), An. barbirostris s.l. (n = 1) and An. vagus (n = 1) were recorded positive for Pf. Anopheles kochi (n = 1) was positive for Pv-210 while An. umbrosus (n = 1), An. nivipes (n = 1) and An. kochi (n = 1) were positive for Pv-247. A mixed infection of Pf and Pv-247 was detected in An. barbirostris s.l.. Conclusion: High diversity of Anopheles species was observed in areas close to the international border where species that were underestimated for malaria transmission significantly outnumbered principal vector species and these may play a significantly heightened role in malaria transmission.

Can the activation of plasminogen/plasmin system of the host by metabolic products of Dirofilaria immitis participate in heartworm disease endarteritis?

Background: Proliferative endarteritis is one of the key pathological mechanisms of cardiopulmonary dirofilariosis, a cosmopolitan parasitosis caused by Dirofilaria immitis affecting dogs and cats around the world. It has been shown that the excretory/secretory antigens from D. immitis adult worms (DiES) bind plasminogen (PLG) and activate fibrinolysis, which can lead to a survival mechanism for the parasite in its intravascular environment. However, overproduction of plasmin (final product of the route) has been related to pathological processes similar to those described in proliferative endarteritis. The aim of this study is to relate the appearance of this pathological condition with the activation of the PLG/plasmin system of the host by DiES. Methods: Cell proliferation through the crystal violet technique, cell migration by wound healing assay and degradation of the extracellular matrix by measuring collagen degradation and levels of matrix metalloproteinases were studied in an “in vitro” model using canine vascular endothelial and smooth muscle cells. These cells were treated with a mixture of DiES + PLG. Untreated cells, cells only stimulated with DiES or with PLG, or with a mixture of DiES + PLG + εACA (an inhibitor of the PLG-plasmin conversion) were employed as controls. In addition, the effect of DiES on the expression of the fibrinolytic activators tPA and uPA, the inhibitor PAI-1 and the PLG receptor Annexin A2 was analyzed in both types of cultures by western blot. Results: Plasmin generated by DiES + PLG binding produced a significant increase in the cell proliferation and migration of the endothelial and smooth muscle cells, as well as an increase in the destruction of the extracellular matrix based on a further degradation of Type I Collagen and an increased level of matrix metalloproteinase-2. DiES also induce an increase in the expression of tPA and uPA in endothelial cells in culture, as well as a decrease in the expression of PAI-1 in both types of cells. Conclusions: Our study reports an interrelationship between plasmin caused by fibrinolysis activation by metabolic products of D. immitis and the appearance of pathological events similar to those described in the emergence of proliferative endarteritis in the cardiopulmonary dirofilariosis.

Genetic structuring and fixed polymorphisms in the gene period among natural populations of Lutzomyia longipalpis in Brazil

Background: Even one hundred years after being originally identified, aspects of the taxonomy of the sand fly Lutzomyia longipalpis, the principal vector of Leishmania infantum in the Americas, remain unresolved for Brazilian populations of this vector. The diversity of morphological, behavioral, biochemical, and ethological characters, as well as the genetic variability detected by molecular markers are indicative of the presence of a complex of species. Methods: In this study, a 525 bp fragment of the period gene was used to evaluate sympatric populations of L. longipalpis. A combination of probabilistic methods such as maximum likelihood and genetic assignment approach to investigate sympatric species of L. longipalpis were applied in three populations of Northeast Brazil. Results: Fixed polymorphism in geographically isolated populations of L. longipalpis from two localities in the state of Ceará and one in the state of Pernambuco, Brazil, was identified in a 525 bp fragment of the gene period(per). Our results suggest a direct relationship between the number of spots found in males’ tergites and the genetic variation in cryptic species of L. longipalpis. The fragment used in this study revealed the nature of the ancestral morphotype 1S. Conclusion: New polymorphisms were identified in the gene per which can be used as a genetic barcode to sympatric taxonomy of L. longipalpis. The per gene fragment confirmed the presence of two siblings species of L. longipalpis in Sobral and showed that these same species are present in two other localities, representing an expansion within the L. longipalpis species complex with regards to the states of Ceará and Pernambuco.

Effect of the physiognomy of Attalea butyracea (Arecoideae) on population density and age distribution of Rhodnius prolixus (Triatominae)

Background: Rhodnius prolixus Stål, 1859 is one of the main vectors of Trypanosoma (Schyzotrypanum) cruzi Chagas, 1909. In its natural forest environment, this triatomine is mainly found in palm tree crowns, where it easily establishes and develops dense populations. The aim of this study was to evaluate the effect of the physiognomy and reproductive status of Attalea butyracea on the population relative density and age structure of R. prolixus and to determine the vector’s population stratification according to the vertical and horizontal profile of an A. butyracea forest. Methods: Using live bait traps, 150 individuals of A. butyracea with different physiognomy and 40 individuals with similar physiognomy (crown size, number of leaves, palm tree height, diameter at breast height, reproductive status) were sampled for triatomines in Yopal, Casanare-Colombia. Temperature and relative humidity were measured in the crown of the palm tree. Entomological indices and natural infection rates were also determined. Results: The relative population density of R. prolixus on natural A. butyracea groves is associated with the palm’s height, number of leaves and crown volume. The young immature stages were present mostly at the crown’s base and the advanced immature stages and adults were present mostly at the crown of the palm tree. This distribution correlates with the temperature stability and relative humidity in the base and the fluctuation of both environmental variables in the palm’s crown. A higher density of R. prolixus was found as the palm tree height increased and as the distance of the palm with respect to the forest border decreased, especially towards anthropically intervened areas. A density index of 12.6 individuals per palm tree with an infestation index of 88.9% and a colonization index of 98.7% was observed. 85.2% was the infection index with T. cruzi. Conclusion: The physiognomy of palm trees affects the relative population density and the distribution of developmental stages of R. prolixus. Therefore, they constitute a risk factor for the potential migration of infected insects from wild environments towards residential environments and the subsequent epidemiological risk of transmission of T. cruzi to people.

Comparison of the performance of three PCR assays for the detection and differentiation of Theileria orientalis genotypes

Background: Oriental theileriosis is a tick-borne disease of bovines caused by the members of the Theileria orientalis complex. Recently, we developed a multiplexed tandem (MT) PCR to detect, differentiate and quantitate four genotypes (i.e., buffeli, chitose, ikeda and type 5) of T. orientalis. In this study, we used MT PCR to assess the prevalence and infection intensity of four T. orientalis genotypes in selected cattle herds that experienced oriental theileriosis outbreaks in New Zealand, and compared the sensitivities and specificities of MT PCR, PCR-high resolution melting (PCR-HRM) and a TaqMan® qPCR. Methods: MT PCR, PCR-HRM analysis for T. orientalis and a TaqMan® qPCR assay for ikeda genotype were employed to test 154 and 88 cattle blood samples from North (where oriental theileriosis outbreaks had occurred; designated as Group 1) and South (where no outbreaks had been reported; Group 2) Islands of New Zealand, respectively. Quantitative data from MT PCR assay were analyzed using generalized linear model and paired-sample t-test. The diagnostic specificity and sensitivity of the assays were estimated using a Bayesian latent class modeling approach. Results: In Group 1, 99.4% (153/154) of cattle were test-positive for T. orientalis in both the MT PCR and PCR-HRM assays. The apparent prevalences of genotype ikeda in Group 1 were 87.6% (134/153) and 87.7% (135/154) using the MT PCR and Ikeda TaqMan® qPCR assays, respectively. Using the MT PCR test, all four genotypes of T. orientalis were detected. The infection intensity estimated for genotype ikeda was significantly higher (P = 0.009) in severely anaemic cattle than in those without anaemia, and this intensity was significantly higher than that of buffeli (P < 0.001) in the former cattle. Bayesian latent class analysis showed that the diagnostic sensitivities (97.1-98.9%) and specificities (96.5-98.9%) of the three PCR assays were very comparable. Conclusion: The present findings show the advantages of using the MT PCR assay as a useful tool for in-depth epidemiological and transmission studies of T. orientalis worldwide.

Invasion of Aedes albopictus (Diptera: Culicidae) into central Africa: what consequences for emerging diseases?

Aedes albopictus, a mosquito native to Asia, has invaded all five continents during the past three decades. It was reported in central Africa in the 2000s, first in Cameroon, and, since then, has colonised almost all countries of the region. The species, originally considered a secondary vector of dengue viruses, has been showed to play a major role in transmission of chikungunya virus in numerous countries, including in the central African region. We review the current spread of Ae. albopictus in central Africa, its larval ecology and its impact on indigenous species such as Ae. aegypti. We explore the potential of Ae. albopictus to affect the epidemiology of emerging or re-emerging arboviruses and discuss the conventional means for its control, while emphasizing the importance of data on its susceptibility to insecticides to cope with potential outbreaks.

Host choice of Phlebotomus orientalis (Diptera: Psychodidae) in animal baited experiments: a field study in Tahtay Adiyabo district, northern Ethiopia

Background: Host choice and feeding success of sand fly vectors of visceral leishmaniasis (VL) are important factors in understanding the epidemiology and for developing efficient control strategies. The aim of the present study was to determine the host preference of Phlebotomus orientalis in the VL focus of Tahtay Adiyabo district, northern Ethiopia. Methods: Two separate experiments were conducted testing attraction to humans, domestic animals, and small wild animals. The host choice of P. orientalis and other sand fly species was assessed using tent traps baited with seven different animals (human, cow, sheep, goat, donkey, dog and chicken) and a blank control. Baited traps were rotated every night in a Latin square design for two consecutive full rounds totaling 16 trap-nights. The second set of experiments tested attraction to small wild animals including; ground squirrel (Xerus rutilus), hare (Lepus sp.), gerbil (Tatera robusta) and spiny mouse (Acomys cahirinus). Animals were caged in standard rodent traps or cylindrical wire-mesh cages. The bait animals were placed in agricultural field and the attracted sand flies were collected using unlit CDC traps for 10 trapping nights. Sand fly specimens collected from each of the experiments were identified to species level and counted. Results: Significant difference (P < 0.05) was observed in the attraction and feeding rate of P. orientalis to different baits. In the first experiment, cow-baited tent traps attracted the highest mean number of P. orientalis (mean = 510 flies). The engorgement rate of P. orientalis on donkey was the highest followed by cow, and much lower on goat, sheep, dog and chicken. In the case of smaller wild animals, more numbers of P. orientalis females were attracted to squirrels followed by hares, gerbils and the spiny rat. However, the engorgement rates for P. orientalis in the smaller animals were very low (1.08%) compared with larger domestic animals (30.53%). Conclusion: The tendency of female P. orientalis to engorge in large numbers on certain species of domestic as well as wild animals strongly indicated that the species is primarily zoophilic in its host preference with feeding habits that may vary depending on the availability of hosts.

Low prevalence of blood parasites in a long-distance migratory raptor: the importance of host habitat

Background: The low prevalence of blood parasites in some bird species may be related to the habitats they frequent, the inexistence of the right host-parasite assemblage or the immunological capacity of the host. Here, we assess the parasite load of breeding populations of Eleonora’s falcon (Falco eleonorae), a medium-sized long-distance migratory raptor that breeds on small isolated islets throughout the Mediterranean basin and overwinters in inland Madagascar. Methods: We examined the prevalence and genetic diversity of the blood parasites belonging to the genera Plasmodium, Haemoproteus and Leucocytozoon in Eleonora’s falcon nestlings from five colonies and in adults from two colonies from nesting sites distributed throughout most of the species’ breeding range. Results: None of the 282 nestlings analysed were infected by blood parasites; on the other hand, the lineages of Plasmodium, Haemoproteus and Leucocytozoon were all found to infect adults. Our results support the idea of no local transmission of vector-borne parasites in marine habitats. Adult Eleonora’s falcons thus may be infected by parasites when on migration or in their wintering areas. Conclusion: The characteristics of marine environments with a lack of appropriate vectors may thus be the key factor determining the absence of local transmission of blood parasites. By comparing the parasite lineages isolated in this species with those previously found in other birds we were able to infer the most likely areas for the transmission of the various parasite lineages.