Parasites and Vectors

Schistosoma mansoni among pre-school children in Musozi village, Ukerewe Island, North-Western-Tanzania: prevalence and associated risk factors

Background: Recent evidence indicates that pre-school children (PSC) living in S. mansoni highly endemic areas are at similar risk of schistosomiasis infection and morbidity as their school aged siblings. Recognizing this fact, the World Health Organization (WHO) is considering including this age group in highly endemic areas in control programmes using mass drug administration (MDA). However, detailed epidemiological information on S. mansoni infection among PSC is lacking for many endemic areas, specifically in Tanzania. This study was conducted to determine the prevalence of S. mansoni infection and its associated risk factors among PSC in Ukerewe Island, North-Western Tanzania. Methods: This was a cross-sectional study, which studied 400 PSC aged 1–6 years. The Kato-Katz (K-K) technique and the point of care circulating cathodic antigen (CCA) immunodiagnostic test were used to diagnose S. mansoni infection in stool and urine samples respectively. A pre-tested questionnaire was used to collect demographic data and water contact behaviour of the children from their parents/guardians. Results: Based on the K-K technique, 44.4 % (95 % CI: 39.4–49.4) pre-school children were infected with S. mansoni and the overall geometric mean eggs per gram of faeces (GM-epg) was 110.6 epg with 38.2 and 14.7 % having moderate and heavy intensity infections respectively. Based on the CCA, 80.1 %, (95 % CI: 76.0–84.0) were infected if a trace was considered positive, and 45.9 %, (95 % CI: 40.9–50.9), were infected if a trace was considered negative. Reported history of lake visits (AOR = 2.31, 95 % CI: 1.06–5.01, P < 0.03) and the proximity to the lake shore (<500 m) (AOR = 2.09, 95 % CI: 1.05–4.14, P < 0.03) were significantly associated with S. mansoni infection. Reported lake visit frequency (4–7 days/week) was associated with heavy intensities of S. mansoni infection (P < 0.00). Conclusion: The prevalence of S. mansoni infection in the study population using K-K and CCA-trace-negative was moderate. The frequency of lake visits and the proximity to the lake shore were associated with the infection of S. mansoni and its intensity. These findings call for the need to include the PSC in MDA programmes, public health education and provision of safe water for bathing.

Humans frequently exposed to a range of non-human primate malaria parasite species through the bites of Anopheles dirus mosquitoes in South-central Vietnam

Background: Recent studies have described natural human infections of the non-human primate parasites Plasmodium knowlesi and Plasmodium cynomolgi. In Southeast Asia, mosquitoes of the Anopheles leucosphyrus group bite both humans and monkeys in the forest and thus offer a possible route for Plasmodium species to bridge the species barrier. In this study we analysed the species composition of malarial sporozoites infecting the salivary glands of Anopheles dirus in order to determine their potential role as bridge vectors of Plasmodium parasites from monkeys to humans. Methods: Mosquitoes were collected in the forest and forest fringe area of Khanh Phu commune by human-baited landing collection. Anopheles species were determined on the basis of morphologic features. Sporozoite-infected salivary glands were applied to filter paper and dried in an ambient atmosphere, before storage in closed vials at 4–6 °C. Detection and identification of Plasmodium species in salivary glands were carried out by nested-PCR of the small subunit ribosomal RNA gene. Results: Six species of Plasmodium parasites were detected by PCR, of which P. vivax was the most common, followed by P. knowlesi, P. inui, P. cynomolgi, P. coatneyi and P. falciparum. Twenty-six of the 79 sporozoite infected mosquitoes showed multiple infections, most of which were a combination of P. vivax with one or more of the non-human primate Plasmodium species. Conclusions: These results suggest that humans overnighting in this forest are frequently inoculated with both human and non-human primate malaria parasites, leading to a situation conducive for the emergence of novel zoonotic malaria.

The formation of egg granulomas in the spleens of mice with late Schistosoma japonicum infection alters splenic morphology

Background: Splenomegaly is a characteristic symptom of schistosome infection. Unlike the well known hepatic pathology of schistosomiasis, splenomegaly has received little scientific research and is generally considered to be a non-specific congestion caused by increased blood pressure within the venous sinuses. Moreover, to date, few studies have reported the deposition of schistosome eggs in the spleen. In a previous study, however, we observed that prolonged S. japonicum infections destroyed the structure of the lymphoid follicles in the spleen of mice at 8 weeks post-infection and found that eggs were frequently deposited in the spleen. These prior observations suggested a relationship between granulomas and splenic morphology which we investigate further in this study. Methods: C57BL/6 mice were infected percutaneously with twenty cercariae of S. japonicum and sacrificed at different times post-infection. The number of eggs present in the homogenates of spleens and livers was quantified by light microscopy. Splenic pathology was observed by immunohistochemistry staining of paraffin-embedded sections. At 18 weeks post-infection the infected mice were divided into two groups (granulomatous spleens and non-granulomatous spleens). Serum antibodies and cytokines in the antigen- or mitogen-stimulated lymphocyte cultures were then determined by ELISA. Results: We found that eggs deposition in the spleens of infected mice occurred frequently but only occasionally led to granulomas formation. The lymphoid follicles within the granulomatous spleens maintained their structural integrity until 20 weeks post-infection, unlike the lymphoid follicles in spleens without egg granulomas. Mice with granulomatous spleens accompanied by lymphoid follicles exhibited a germinal center (GC)-like structure and had enhanced humoral immune responses. Splenocytes from granulomatous spleens also showed significantly elevated levels of Th2 cytokines during late infection stages. Conclusions: Our results highlight that lymphoid follicles, which are not completely destroyed or are re-established in the spleen, can change the local immune environment and lead to changes in the splenic morphology of mice with chronic schistosomiasis.

Not every worm wrapped around a stick is a guinea worm: a case of Onchocerca volvulus mimicking Dracunculus medinensis

Background: Despite being certified guinea worm free in 2007, Cameroon continues surveillance efforts to ensure rapid verification of any suspected reoccurrence. This includes the investigation of every rumor and confirmation of each suspicious expulsed worm. This paper presents fieldwork carried out to investigate a guinea worm rumor in Cameroon which turned out to be an Onchocerca volvulus mimicking Dracunculus medinensis. Methods: The investigation included a field visit to the subsistence farming community where the rumor was reported. During the visit, interviews were conducted with health staff who managed the case and the elderly farmer from whom the worm was retrieved. An investigation of any potential missed guinea worm cases was also conducted through interviews with community residents and reviews of the health facility’s medical records. This was combined with laboratory analyses of water samples from the community’s water sources and the retrieved worm which was removed from the patient via wrapping it around a stick. Results: Microscopy and molecular analyses of the retrieved worm revealed a female Onchocerca volvulus whose expulsion strongly mimicked guinea worm. In addition to presenting findings of our investigation, this paper discusses distinguishing elements between the two parasites and gives an overview of guinea worm eradication efforts in Cameroon as well as current challenges to the worm’s eradication globally. Conclusions: The investigation findings suggest the evolving Onchocerca volvulus worm tropisms’ adaptive survival behavior worth further investigation. Strategies used to successfully control guinea worm in Cameroon could be adapted for Onchocerca volvulus control.

In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus

Background: Culex quinquefasciatus is a hematophagous insect from the Culicidae family that feeds on the blood of humans, dogs, birds and livestock. This species transmits a wide variety of pathogens between humans and animals. The midgut environment is the first location of pathogen-vector interactions for blood-feeding mosquitoes and the expression of specific peptidases in the early stages of feeding could influence the outcome of the infection. Trypsin-like serine peptidases belong to a multi-gene family that can be expressed in different isoforms under distinct physiological conditions. However, the confident assignment of the trypsin genes that are expressed under each condition is still a challenge due to the large number of trypsin-coding genes in the Culicidae family and most likely because they are low abundance proteins. Methods: We used zymography for the biochemical characterization of the peptidase profile of the midgut from C. quinquefasciatus females fed on sugar. Protein samples were also submitted to SDS-PAGE followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis for peptidase identification. The peptidases sequences were analyzed with bioinformatics tools to assess their distinct features. Results: Zymography revealed that trypsin-like serine peptidases were responsible for the proteolytic activity in the midgut of females fed on sugar diet. After denaturation in SDS-PAGE, eight trypsin-like serine peptidases were identified by LC-MS/MS. These peptidases have structural features typical of invertebrate digestive trypsin peptidases but exhibited singularities at the protein sequence level such as: the presence of different amino acids at the autocatalytic motif and substrate binding regions as well as different number of disulfide bounds. Data mining revealed a group of trypsin-like serine peptidases that are specific to C. quinquefasciatus when compared to the culicids genomes sequenced so far. Conclusion: We demonstrated that proteomics approaches combined with bioinformatics tools and zymographic analysis can lead to the functional annotation of trypsin-like serine peptidases coding genes and aid in the understanding of the complexity of peptidase expression in mosquitoes.

Can human schistosomiasis mansoni control be sustained in high-risk transmission foci in Egypt?

Background: Control of human schistosomiasis remains a longstanding issue on the agenda of the Egyptian Ministry of Health and Population (MOHP). Substantial impact on morbidity and prevalence of S. mansoni was widely reported after the National Schistosomiasis Control Program (NSCP) extended selective treatment with praziquantel (PZQ) to the Nile Delta in 1992 and upgrading this approach to mass drug administration (MDA) in 1997. Disease elimination, however, eludes NSCP as the micro-level includes many high-risk foci that sustain transmission, which has not been subjected to investigation. Methods: The study included five high-risk Nile Delta villages situated in the Kafr El-Sheikh Governorate. The total sample size amounted to 2382 individuals of both sexes and all ages. Diagnosis was based on four Kato-Katz slides from two consecutive stool samples. Data were investigated using SPSS, comparing proportions with the Chi square test and means with the Student t test, while strength of the associations were subjected to Odds Ratio (OR) analysis. Results: The overall prevalence of schistosomiasis in the study area was found to be 29 %, while the mean geometric mean egg count (GMEC) was low (66.78 ± 4.4) indicating low intensity of infection. The mean village prevalence rates ranged from 16.5 % to 49.5 % and the GMECs from 35.2 to 86.2 eggs per gram (EPG) of stool. The difference of prevalence between villages was statistically significant at P < 0.05, and the prevalence was significantly higher among males than among females, P < 0.05, OR =1.4 and 95 % CI (1.16-1.60). Infection peaked in the next youngest age group (5- ≤ 10 years of age) at an average prevalence of 50.8 % with the GMEC reaching 209 EPG of stool in the village with the highest prevalence. The average prevalence and GMEC among children <5 years were 20.6 % and 92.7 EPG, respectively. Conclusion: Transmission of S mansoni in high-risk areas in the Nile Delta remains uninterrupted calling for improved, more comprehensive control strategies. Further investigations are needed to find out whether these results are due to inefficacy of PZQ, surviving immature worms or drug resistance.

Bacteria-induced egg hatching differs for Trichuris muris and Trichuris suis

Background: Eggs of the porcine whipworm Trichuris suis are currently explored in human clinical trials as a treatment of immune-mediated diseases. In this context, only the infective, embryonated eggs, constitute the Active Pharmaceutical Ingredient (API). The rodent whipworm, Trichuris muris is commonly used as a laboratory model to study Trichuris biology. The embryonated eggs (containing a fully developed larva) are biologically active and will invade the large intestinal mucosa of the host. This study aims to assess the in vitro hatching of T. muris and T. suis eggs in various bacterial cultures as a measure for their biological activity. Methods: Eggs of T. muris and T. suis were incubated with Escherichia coli strain (BL-21) at three concentrations in a slightly modified in vitro egg hatching assay previously developed for T. muris. Additionally, E. coli strains (M15, SG13009, PMC103, JM109, TUNER, DH5alpha, TOP10) and five Gram-positive bacteria (Enterococcus caccae, Streptococcus hyointestinalis, Lactobacillus amylovorus, L. murinus, and L. reuteri) were tested as a hatching stimulus for T. muris and T. suis eggs. Results: Whereas T. muris eggs hatched, T. suis did not, even when exposed to different concentrations and strains of E. coli after 4 and 24-hour incubation. When incubated with Gram-positive bacteria, only T. muris eggs showed noticeable hatching after 20 h, although with high variability. Conclusions: The observed difference in hatching of T. muris and T. suis eggs incubated with selected bacteria, indicate significant biological differences which may reflect specific adaptation to different host-specific gut microbiota.

Development of a LAMP assay for detection of Leishmania infantum infection in dogs using conjunctival swab samples

Background: Leishmania infantum infections in dogs play a crucial role in the transmission of pathogens causing visceral leishmaniasis to humans in the Gansu province, northwest China. To be able to control zoonotic transmission of the parasite to humans, a non-invasive loop-mediated isothermal amplification (LAMP) assay to specifically detect L. infantum infections in dogs was developed. Methods: The primers used in the LAMP assay were designed to target kinetoplast DNA minicircle sequences of the L. infantum isolate MCAN/CN/90/SC and tested using DNA isolated from promastigotes of different Leishmania species. The LAMP assay was evaluated with conjunctional swab samples obtained from 111 and 33 dogs living in an endemic and a non-endemic region of zoonotic visceral leishmaniasis in the Gansu province, respectively. The LAMP assay was also compared with conventional PCR, ELISA and microscopy using conjunctional swab, serum and bone marrow samples from the dogs, respectively. Results: The LAMP assay detected 1 fg of L. infantum DNA purified from cultured promastigotes which was 10-fold more sensitive than a conventional PCR test using Leishmania genus-specific primers. No cross reaction was observed with DNA isolated from promastigotes of L. donovani, L. major, L. tropica, and L. braziliensis, and the L. infantum reference strain MHOM/TN/80/IPT1. The L. infantum-positive rates obtained for field-collected samples were 61.3 %, 58.6 %, 40.5 % and 10.8 % by LAMP, PCR, ELISA and microscopy, respectively. As only one out of the 33 samples from control dogs from the non-endemic region of zoonotic visceral leishmaniasis was positive by the LAMP assay and the PCR test, the observed true negative rate (specificity) was 97 % for both methods. Conclusion: This study has shown that the non-invasive, conjunctional swab-based LAMP assay developed was more sensitive in the detection of leishmaniasis in dogs than PCR, ELISA and microscopy. The findings indicate that the LAMP assay is a sensitive and specific method for the field surveillance of domestic dogs, particularly of asymptomatic canines, in ZVL-endemic areas in western China.

A comparison of two tests for filarial antigenemia in areas in Sri Lanka and Indonesia with low-level persistence of lymphatic filariasis following mass drug administration

Background: Filarial antigen tests are key tools for mapping the distribution of bancroftian filariasis and for detecting areas with persistent infections following mass drug administration (MDA). A recent study showed that the new Alere Filariasis Test Strip (FTS) has better analytical sensitivity than the BinaxNOW Filariasis card test (Card Test) for detecting circulating filarial antigen, and the FTS detected more positive results than the Card Test in a field study performed in a highly endemic area in Liberia. Methods: The present study compared the performance of the FTS and the Card Test in community surveys that were conducted in southern Sri Lanka and in Indonesia (Central Java) in areas with low-level persistence of LF following multiple rounds of MDA with diethylcarbamazine plus albendazole. The studies were performed in densely populated semi-urban areas where Wuchereria bancrofti is transmitted by Culex quinquefasciatus. Results: Antigenemia rates by FTS were 138 % higher in the Sri Lanka study (43/852 vs. 18/852) and 21 % higher in the Indonesia study (50/778 vs. 41/778) than antigenemia rates by Card Test. Antigenemia rates were significantly higher in males than in females and higher in adults than in children in both study sites. Although overall antigenemia rates and test scores were significantly higher by FTS than by Card Test in both study areas, rates in young children were similar with both tests in both areas. Conclusions: These results extend the previously reported superior sensitivity of the FTS to areas with low residual infection rates following MDA, and this could affect mapping and post-MDA survey results in adults. However, our findings suggest that results of transmission assessment surveys (TAS) performed in school-aged children are likely to be similar with both tests.

Identification of genes associated with blood feeding in the cat flea, Ctenocephalides felis

Background: The cat flea (Ctenocephalides felis) is a blood-feeding ectoparasitic insect and particular nuisance pest of companion animals worldwide. Identification of genes that are differentially expressed in response to feeding is important for understanding flea biology and discovering targets for their control. Methods: C. felis fleas were maintained and fed for 24 h using an artificial rearing system. The technique of suppression subtractive hybridization was employed to screen for mRNAs specifically expressed in fed fleas. Results: We characterized nine distinct full-length flea transcripts that exhibited modulated or de novo expression during feeding. Among the predicted protein sequences were two serine proteases, a serine protease inhibitor, two mucin-like molecules, a DNA topoisomerase, an enzyme associated with GPI-mediated cell membrane attachment of proteins and a component of the insect innate immune response. Conclusions: Our results provide a molecular insight into the physiology of flea feeding. The protein products of the genes identified may play important roles during flea feeding in terms of blood meal digestion, cellular growth/repair and protection from feeding-associated stresses.

Identifying biotic interactions which drive the spatial distribution of a mosquito community

Background: Spatial variation in the risk of many mosquito-borne pathogens is strongly influenced by the distribution of communities of suitable vector mosquitoes. The spatial distributions of such communities have been linked to the abiotic habitat requirements of each constituent mosquito species, but the biotic interactions between mosquitoes and other species are less well understood. Determining which fauna restrict the presence and abundance of key mosquito species in vector communities may identify species which could be employed as natural biological control agents. Whilst biotic interactions have been studied in the laboratory, a lack of appropriate statistical methods has prohibited the identification of key interactions which influence mosquito distributions in the field. Joint species distribution models (JSDMs) have recently been developed to identify biotic interactions influencing the distributions of species from empirical data. Methods: We apply a JSDM to field data on the spatial distribution of mosquitoes in a UK wetland to identify both abiotic factors and biotic interactions driving the composition of the community. Results: As expected, mosquito larval distributions in this wetland habitat are strongly driven by environmental covariates including water depth, temperature and oxidation-reduction potential. By factoring out these environmental variables, we are able to identify species (ditch shrimp of the genus Palaemonetes and fish) as predators which appear to restrict mosquito distributions. Conclusions: JSDMs offer vector ecologists a way to identify potentially important biotic interactions influencing the distributions of disease vectors from widely available field data. This information is crucial to understand the likely effects of habitat management for vector control and to identify species with the potential for use in biological control programmes. We provide an R package BayesComm to enable the wider application of these models.

The effect of synthetic pyrethroids on the attachment and host-feeding behaviour in Dermacentor reticulatus females (Ixodida: Amblyommidae)

Background: The high competence of D. reticulatus in transmission of tick-borne pathogens prompts investigations of the effect of chemicals used as repellents and acaricides on the behaviour of the tick on the host. Therefore, this paper presents the effect of permethrin and deltamethrin on the attachment and feeding in this tick species.FindingsAttachment to rabbit skin of D. reticulatus females sprayed with pyrethroids and the effect of different doses thereof on feeding were assessed at a temperature of 20 ± 3 °C and 50 % humidity.The dynamics of attachment of D. reticulatus females varied in a dose-dependent manner after the application of both pyrethroids. Within the first 0.5 h of the experiments, there was an over six-fold and over twelve-fold increase in the number of females attached to host skin after application of permethrin concentrations of 0.3906–0.7812 μg and 1.5625–3.1250 μg/1 specimen, respectively. In the case of deltamethrin, females treated with the dose of 0.0390 μg of the compound were able to attach to host skin only 4 hours after the infestation.The toxic activity of both pyrethroids increased the duration of the feeding period and decreased the body weight of engorged females and the feeding efficiency index. Conclusions: The accelerated attachment of D. reticulatus females caused by sublethal permethrin doses and delayed or inhibited attachment caused by deltamethrin suggest a necessity of careful selection of the type and dose of pyrethroids to protect hosts from tick attacks.

Identifying avian malaria vectors: sampling methods influence outcomes

Background: The role of vectors in the transmission of avian malaria parasites is currently understudied. Many studies that investigate parasite-vector relationships use limited trapping techniques and/or identify potential competent vectors in the field in such ways that cannot distinguish between an infected or infectious vector. Without the use of multiple trapping techniques that address the specific biology of diverse mosquito species, and without looking at the infection status of individual mosquitoes, it is not possible to make dependable conclusions on the role of mosquitoes in the transmission of avian malaria parasites. Methods: We conducted two years of mosquito collections at a riparian preserve in California where a wide diversity of species were collected with multiple trap types. We hypothesized that competent mosquito species can influence the distribution and diversity of avian malaria parasites by acting as a compatibility filter for specific Plasmodium species. To determine the infection status of all individual mosquitoes for Plasmodium species/lineages, amplification within the cytochrome b gene was carried out on over 3000 individual mosquito thoraxes, and for those that tested positive we then repeated the same process for abdomens and salivary glands. Results: Our data show heterogeneity in the transmissibility of Plasmodium among ornithophillic mosquito species. More specifically, Culex stigmatosoma appears to not be a vector of Plasmodium homopolare, a parasite that is prevalent in the avian population, but is a vector of multiple other Plasmodium species/lineages. Conclusions: Our results suggest that conclusions made on the role of vectors from studies that do not use different mosquito trapping methods should be re-evaluated with caution, as we documented the potential for trapping biases, which may cause studies to miss important roles of specific mosquito species in the transmission of avian malaria. Moreover, we document heterogeneity in the transmission of Plasmodium spp. by mosquitoes can influence Plasmodium diversity and prevalence in specific locations to Plasmodium-vector incompatibilities.

Efficacy of fluralaner flavored chews (Bravecto&#174;) administered to dogs against the adult cat flea, Ctenocephalides felis felis and egg production

Background: Fluralaner is a potent insecticide and acaricide with rapid and persistent efficacy. This study measured the efficacy of fluralaner flavored chews (Bravecto®, Merck Animal Health) administered to dogs against adult Ctenocephalides felis felis and egg production. Methods: Twelve purpose-bred dogs were randomly allocated to two groups of six dogs each. Dogs in treatment group 1 were administered a single fluralaner flavored chew to achieve a minimum dose of at least 25 mg/kg while treatment group 2 served as untreated controls. On Days −2, 28, 56, 84, 91, 98, 105, 112, and 120 post-treatment, each dog was infested with approximately 200 unfed cat fleas, C. felis felis (KS1 strain). Forty-eight hours after treatment and 48 h after each infestation, eggs were collected over a 3-h period, counted and viability determined. Dogs were combed to remove any remaining fleas. Results: Treatment of dogs with oral fluralaner provided a 100 % reduction in flea counts 48 h after treatment and within 48 h of every post-treatment infestation through Day122. Egg production from fluralaner treated dogs was reduced by 99.9 % (two eggs from one dog) within 48 h after treatment and not a single egg (100 % efficacy) was thereafter collected from treated dogs. Adult flea counts and egg production from the fluralaner-treated dogs were significantly lower than for non-treated controls at all post-treatment evaluations (P < 0.001). The two eggs collected from the single treated dog 48 h after treatment did not produce any adult fleas. As no additional eggs were collected from treated dogs, no viability assessment was performed. Conclusions: A single oral dose of fluralaner flavored chews provided 100 % efficacy against repeated flea infestations on dogs for 4 months. Fluralaner reduced egg production of activity reproducing female fleas by 99.9 % and then killed every single female flea before any eggs could be produced following each subsequent re-infestation for the entire 122-day evaluation period.

A Leishmania -specific hypothetical protein expressed in both promastigote and amastigote stages of Leishmania infantum employed for the serodiagnosis of, and as a vaccine candidate against, visceral leishmaniasis

Background: LiHyV is an antigenic hypothetical protein present in both promastigote and amastigote stages of Leishmania infantum, which was recently identified by an immunoproteomic approach. A recombinant version of this protein (rLiHyV) was evaluated as a diagnostic marker for canine VL (CVL). In addition, the prophylactic efficacy of the rLiHyV protein, and two of its CD8 + T cell epitopes, has been analyzed in a murine model of visceral leishmaniasis (VL). Methods: Initially, the rLiHyV protein was evaluated by an ELISA technique for the serodiagnosis of CVL. Secondly, vaccines composed of the recombinant protein and both chemically synthesized peptides, combined with saponin as an adjuvant; were administered subcutaneously into BALB/c mice. The cellular and humoral responses generated by vaccination were evaluated. In addition, the parasite burden and immune response were studied 10 weeks after L. infantum infection. Results: The rLiHyV protein was recognized by antibodies of VL dogs. No cross-reactivity was obtained with sera from dogs vaccinated with a Brazilian commercial vaccine, with sera from animals infected with Trypanosoma cruzi, Babesia canis and Ehrlichia canis, or those from non-infected animals living in an endemic area for leishmaniasis. After challenge with L. infantum, spleen cells of BALB/c mice vaccinated with rLiHyV/saponin stimulated with parasite antigens showed a higher production of IFN-γ, IL-12 and GM-CSF, than the same cells obtained from mice vaccinated with the individual peptides, or mice from control (inoculated with saline or saponin) groups. This Th1-type cellular response observed in rLiHyV/saponin vaccinated mice was accompanied by the induction of parasite-specific IgG2a isotype antibodies. Animals immunized with rLiHyV/saponin showed significant reductions in the parasite burden in the liver, spleen, bone marrow and in the lymph nodes draining the paws relative to control mice. Conclusions: The present study showed for the first time that the L. infantum LiHyV protein could be considered as a vaccine candidate against L. infantum infection, as well as a diagnostic marker for CVL.

Efficacy of praziquantel against urinary schistosomiasis and reinfection in Senegalese school children where there is a single well-defined transmission period

Background: Human schistosomiasis is a significant health problem in Sub-Saharan Africa. In Niakhar, West central Senegal, the transmission of S. haematobium occurs seasonally between July and November. No control measures have been implemented despite high prevalence reported in previous studies. This aim of this study was to i) determine the current prevalence of S. haematobium in children at Niakhar, ii) assess the efficacy of one dose of PZQ (40 mg/kg) against S. haematobium and iii) monitor reinfection. Methods: The current study was carried out in a cohort of 329 children aged five to 15 years enrolled from six villages in Niakhar to determine the efficacy of one dose of PZQ, as well as reinfection. Parasitological screening was performed in June 2011 to determine the baseline prevalence of S. haematobium, and then a single dose of PZQ was administered to all selected subjects in the transmission season in August 2011. The efficacy of PZQ treatment and reinfection were monitored respectively five weeks after in September 2011 and from February to March 2012. Results: At baseline, the overall prevalence and the heavy intensity of infection were 73.2 % and 356.1eggs/10 ml of urine. Significant differences in the prevalence and intensity of S. haematobium infection were noted between villages. A single dose of PZQ significantly reduced the prevalence of S. haematobium infection from 73.2 % to 4.6 % and the geometric mean intensity of infection from 356.1 to 43.3 eggs/10 ml of urine. The cure rates ranged from 89.4 % to 100 %. The egg reduction rates also ranged from 77.6 % to 100 %. Two to three months after the period of transmission, the overall rate of reinfection was 12.6 % and was significantly higher in male children than in female children. The overall prevalence at this period was 13.8 %, which was significantly lower than the prevalence at baseline (73.2 %). Conclusion: The Niakhar study area remains a hot spot of urinary schistosomiasis in Senegal with differences in transmission between villages. This study suggests that when transmission is strictly seasonal, Praziquantel shows the expected efficacy in reducing the prevalence and intensity of infection, but also a significant effect on the occurrence of reinfection.

Does Triatoma brasiliensis occupy the same environmental niche space as Triatoma melanica ?

Background: Triatomines (Hemiptera, Reduviidae) are vectors of Trypanosoma cruzi, the causative agent of Chagas disease, one of the most important vector-borne diseases in Latin America. This study compares the environmental niche spaces of Triatoma brasiliensis and Triatoma melanica using ecological niche modelling and reports findings on DNA barcoding and wing geometric morphometrics as tools for the identification of these species. Methods: We compared the geographic distribution of the species using generalized linear models fitted to elevation and current data on land surface temperature, vegetation cover and rainfall recorded by earth observation satellites for northeastern Brazil. Additionally, we evaluated nucleotide sequence data from the barcode region of the mitochondrial cytochrome c oxidase subunit I (CO1) and wing geometric morphometrics as taxonomic identification tools for T. brasiliensis and T. melanica. Results: The ecological niche models show that the environmental spaces currently occupied by T. brasiliensis and T. melanica are similar although not equivalent, and associated with the caatinga ecosystem. The CO1 sequence analyses based on pair wise genetic distance matrix calculated using Kimura 2-Parameter (K2P) evolutionary model, clearly separate the two species, supporting the barcoding gap. Wing size and shape analyses based on seven landmarks of 72 field specimens confirmed consistent differences between T. brasiliensis and T. melanica. Conclusion: Our results suggest that the separation of the two species should be attributed to a factor that does not include the current environmental conditions. However, as the caatinga is a biome that has existed in the area for at least the last 18,000 years, past conditions might have had an influence in the speciation process. The DNA Barcoding approach may be extended to these members of the subfamily Triatominae.

Exposure to Leishmania spp. and sand flies in domestic animals in northwestern Ethiopia

Background: Human visceral leishmaniasis caused by Leishmania donovani is considered an anthroponosis; however, Leishmania-infected animals have been increasingly reported in L. donovani foci, and the role of these animals as reservoirs for human L. donovani infection remains unclear. Methods: We conducted a study of domestic animals (goats, sheep, cows, dogs, and donkeys) in three L. donovani foci in northwestern Ethiopia. Domestic animals were screened for Leishmania DNA and for anti-L. donovani IgG. Serum anti-sand fly saliva antibodies were used as a marker of exposure to the vector sand fly, Phlebotomus orientalis. Results: Of 546 animals tested, 32 (5.9 %) were positive for Leishmania DNA, with positive animals identified among all species studied. Sequencing indicated that the animals were infected with parasites of the L. donovani complex but could not distinguish between L. infantum and L. donovani. A total of 18.9 % of the animals were seropositive for anti-L. donovani IgG, and 23.1 % of the animals were seropositive for anti-P. orientalis saliva IgG, with the highest seroprevalence observed in dogs and sheep. A positive correlation was found between anti-P. orientalis saliva and anti-L. donovani IgGs in cows, goats, and sheep. Conclusions: The detection of L. donovani complex DNA in the blood of domestic animals, the reported seroprevalence to the L. donovani antigen, and the widespread exposure to sand fly saliva among domestic animals indicate that they are frequently exposed to Leishmania infection and are likely to participate in the epidemiology of Leishmania infection, either as potential blood sources for sand flies or possibly as parasite hosts.

Enteric neuromodulators and mucus discharge in a fish infected with the intestinal helminth Pomphorhynchus laevis

Background: In vertebrates, the presence of enteric worms can induce structural changes to the alimentary canal impacting on the neuroendocrine system, altering the proper functioning of the gastrointestinal tract and affecting the occurrence and relative density of endocrine cells (ECs). This account represents the first immunohistochemistry and ultrastructure-based study which documents the intimate relationship between the intestinal mucous cells and ECs in a fish-helminth system, investigating the potential effects of enteric neuromodulators on gut mucus secretion/discharge. Methods: A modified dual immunohisto- and histochemical staining technique was applied on intestinal sections from both infected and uninfected fish. Sections were incubated in antisera to a range of neuromodulators (i.e. leu-enkephalin, met-enkephalin, galanin and serotonin) and the glycoconjugate histochemistry of the mucous cells was determined using a subsequent alcian blue – periodic acid Schiff staining step. Dual fluorescent staining on sections prepared for confocal laser scanning microscopy and transmission electron microscopy were also used to document the relationship between ECs and mucous cells. Results: From a total of 26 specimens of Squalius cephalus sampled from the River Paglia, 16 (i.e. 62 %) specimens were found to harbour an infection of the acanthocephalan Pomphorhynchus laevis (average intensity of infection 9.2 ± 0.8 parasites host −1 , mean ± standard error). When acanthocephalans were present, the numbers of mucous cells (most notably those containing acidic or mixed glycoconjugates) and ECs secreting leu-enkephalin, met-enkephalin, galanin, serotonin were significantly higher than those seen on sections from uninfected fish. The relationship between met-enkephalin-like or serotonin-like ECs and lectin DBA positive mucous cells was demonstrated through a dual fluorescent staining. The presence of tight connections and desmosomes between mucous and ECs in transmission electron micrographs provides further evidence of this intimate relationship. Conclusions: The presence of P. laevis induces an increase in the number of enteric ECs that are immunoreactive to leu- and met-enkephalin, galanin, and serotonin anti-sera. The mucous cells hyperplasia and enhanced mucus secretion in the helminth-infected intestines could be elicited by the increase in the number of ECs which release these regulatory substances.