Background: Transmission of pathogens by ticks is greatly supported by tick saliva released during feeding. Dendritic cells (DC) act as immunological sentinels and interconnect the innate and adaptive immune system. They control polarization of the immune response towards Th1 or Th2 phenotype. We investigated whether salivary cystatins from the hard tick Ixodes scapularis, sialostatin L (Sialo L) and sialostatin L2 (Sialo L2), influence mouse dendritic cells exposed to Borrelia burgdorferi and relevant Toll-like receptor ligands. Methods: DCs derived from bone-marrow by GM-CSF or Flt-3 ligand, were activated with Borrelia spirochetes or TLR ligands in the presence of 3 μM Sialo L and 3 μM Sialo L2. Produced chemokines and IFN-β were measured by ELISA test. The activation of signalling pathways was tested by western blotting using specific antibodies. The maturation of DC was determined by measuring the surface expression of CD86 by flow cytometry. Results: We determined the effect of cystatins on the production of chemokines in Borrelia-infected bone-marrow derived DC. The production of MIP-1α was severely suppressed by both cystatins, while IP-10 was selectively inhibited only by Sialo L2. As TLR-2 is a major receptor activated by Borrelia spirochetes, we tested whether cystatins influence signalling pathways activated by TLR-2 ligand, lipoteichoic acid (LTA). Sialo L2 and weakly Sialo L attenuated the extracellular matrix-regulated kinase (Erk1/2) pathway. The activation of phosphatidylinositol-3 kinase (PI3K)/Akt pathway and nuclear factor-κB (NF-κB) was decreased only by Sialo L2. In response to Borrelia burgdorferi, the activation of Erk1/2 was impaired by Sialo L2. Production of IFN-β was analysed in plasmacytoid DC exposed to Borrelia, TLR-7, and TLR-9 ligands. Sialo L, in contrast to Sialo L2, decreased the production of IFN-β in pDC and also impaired the maturation of these cells. Conclusions: This study shows that DC responses to Borrelia spirochetes are affected by tick cystatins. Sialo L influences the maturation of DC thus having impact on adaptive immune response. Sialo L2 affects the production of chemokines potentially engaged in the development of inflammatory response. The impact of cystatins on Borrelia growth in vivo is discussed.

Background: Cyclospora is a protistan parasite that causes enteritis in several species of animals including humans. The aim of this study was to investigate the presence of Cyclospora in captive non-human primates. Methods: A total of 119 faecal samples from Pan troglodytes, Macaca sylvanus, Cercopithecus cephus, Erythrocebus patas, Chlorocebus aethiops, and Macaca fascicularis from a wildlife animal rescue center, as well as from Macaca fascicularis from an experimental primate research center were tested for the presence of Cyclospora by quantitative real time PCR (qPCR) and single-strand conformation polymorphism (SSCP) analysis. Results: Cyclospora was detected in three Pan troglodytes (13.6%) and nine (9.3%) Macaca fascicularis. Conclusions: The present study represents the first record of Cyclospora in captive primates in Europe, suggesting the presence of Cyclospora cayetanensis, which is transmissible to humans.

Background: Thelazia callipaeda is transmitted by Phortica variegata, a drosophilid that feeds on lachrymal secretions of mammals. Scientific information on human thelaziosis is still relatively limited, mainly for physicians and ophthalmologists. Indeed, the literature is full of misleading information on the transmission of T. callipaeda to humans.FindingsA recent paper reported a case of human intraocular infestation in a patient from Karnataka. The information presented in that article as well as in other articles in the international literature is outdated and incorrect in several instances, mostly regarding to the localization of T. callipaeda in the host, its biology and routes of transmission. Conclusions: Physicians and ophthalmologists should be aware that T. callipaeda is larviparous and transmitted exclusively by secretophagous flies. These flies buzz around the eyes of animals and humans at the daytime, landing on the eyes and releasing the infective larvae on the host conjunctiva. That is the only possible way of transmission of T. callipaeda.

Background: Dengue is a prevalent arboviral disease and the development of insecticide resistance among its vectors impedes endeavors to control it. Coffee is drunk by millions of people daily worldwide, which is associated with the discarding of large amounts of waste. Coffee and its waste contain large amounts of chemicals many of which are highly toxic and none of which have a history of resistance in mosquitoes. Once in solution, coffee is brownish in colour, resembling leaf infusion, which is highly attractive to gravid mosquitoes. To anticipate the environmental issues related to the increasing popularity of coffee as a drink, and also to combat insecticide resistance, we explored the deterrence potentials of coffee leachates against the ovipositing and embryonic stages of the dengue vector, Aedes albopictus. Methods: In a series of choice, no-choice, and embryo toxicity bioassays, we examined changes in the ovipositional behaviours and larval eclosion of Ae. albopictus in response to coffee extracts at different concentrations. Results: Oviposition responses were extremely low when ovicups holding highly concentrated extract (HCE) of coffee were the only oviposition sites. Gravid females retained increased numbers of mature eggs until 5 days post-blood feeding. When provided an opportunity to oviposit in cups containing coffee extracts and with water, egg deposition occurred at lower rates in those containing coffee, and HCE cups were far less attractive to females than those containing water only. Females that successfully developed in a coffee environment preferentially oviposited in such cups when in competition with preferred oviposition sites (water cups), but this trait did not continue into the fourth generation. Larval eclosion occurred at lower rates among eggs that matured in a coffee environment, especially among those that were maintained on HCE-moistened substrates. Conclusions: The observations of the present study indicate a pronounced vulnerability of Ae. albopictus to the presence of coffee in its habitats during the early phases of its life cycle. The observations that coffee repels gravid females and inhibits larval eclosion provide novel possibilities in the search for novel oviposition deterrents and anti-larval eclosion agents against dengue vectors.

This letter advises the imminent formation of the Companion Animal Parasites Council for the Tropics (CAPCT). The CAPCT consists of region-specific (e.g., Asia-Pacific, Latin America and Caribbean, Africa) experts comprising academics, veterinarians, parasitologists, physicians and allied industry partners that will work together to inform, guide and develop best-practice recommendations for the optimal diagnosis, treatment and control of companion animal parasites in the tropics, with the aim of protecting the health of pets and that of the public.

Background: Blood-feeding behavior studies are important for estimating the efficiency of pathogen transmission and assessing the relative human disease risk. However, in Ethiopia and other parts of East Africa there are large remaining gaps in identifying the feeding habits of Phlebotomus orientalis, the vector of Leishmania donovani. The aim of the study was to determine the blood feeding patterns of P. orientalis in Tahtay Adiyabo district, northern Ethiopia. Methods: For bloodmeal analysis, sandflies were collected from three different villages of Tahtay Adiyabo district using CDC light traps, sticky traps, and pyrethrum spray catches. Bloodmeal of engorged female sandflies was identified using cytochrome (cyt) b-PCR and reverse-line blotting (RLB) and enzyme linked immunosorbent assay (ELISA) assays. Results: Most (637/641) of the females analyzed were P. orientalis. Successful identification of the host from bloodmeals was achieved in 83.03 and 92.1 % using cyt b PCR-RLB and ELISA, respectively. Bloodmeal analysis of P. orientalis females revealed that they have a range of hosts with predominant preference to bovines followed by donkey, human, goat, sheep, dog, and camel. Conclusion: Results obtained from bloodmeal analyses demonstrate that the feeding preference of P. orientalis is mainly zoophilic, which could vary depending on the availability of hosts.

Background: Triatoma sordida, a vector of Chagas disease, is a native of Brazil, Bolivia, Paraguay, Argentina, and Uruguay, and occurs primarily in peridomiciles. Currently, it is the species most frequently captured by the Chagas Disease Control Program in Brazil. For this reason, population genetic studies attract great interest, as they can provide further information about the dispersal and household invasion processes of this species. In the absence of suitable markers, the objective of this study was to test the cross amplification of microsatellite primers.Findings23 primers were tested for microsatellite loci already described for other species of the genus Triatoma sp. Forty four specimens of T. sordida captured in the north of Minas Gerais were used to validate the use of standardized loci for population genetic analyses. It was possible to amplify 10 of the 23 loci tested for T. sordida. Conclusions: This is the first study that provides 10 microsatellite markers for population analysis of this triatomine species. Cross-amplification of primers can be used among other phylogenetically related species whose loci are already available for study.

Background: Most argasid ticks from the Neotropical region are parasites of mammals and birds, with a few records from reptiles. Many species of the genus Ornithodoros are known only through larval descriptions, and their chaetotaxy and morphological characteristics have been used to separate the taxa. In the present study, we describe the larva and the nymph of first instar of a new species of the genus Ornithodoros that was collected from frogs of the species Thoropa miliaris. Methods: Larvae of Ornithodoros were collected from frogs of the species T. miliaris at waterfalls in the state of Rio de Janeiro, southeastern Brazil. The larval and nymphal description was based on optical and scanning electron microscopy. Molecular analysis using the argasid 16S rRNA sequences available in GenBank was also conducted. Results: Ornithodoros faccinii sp. n. is closely related to Ornithodoros clarki Jones & Clifford, Ornithodoros marinkellei Kohls, Clifford & Jones, Ornithodoros capensis Neumann and Ornithodoros sawaii Kitaoka & Susuki. However, the larval morphology of the new species is unique. The mitochondrial 16S rDNA partial sequence of O. faccinii generated in the present study was deposited in GenBank under the number KP861242. Conclusions: The larvae collected from Thoropa miliaris are a new species, Ornithodoros faccinii n. sp. This is the first report of argasid ticks on frogs in Brazil, the second on frogs and the third on Amphibia in the Neotropical region.

Background: Although the burden of malaria has significantly declined in recent years in sub-Saharan Africa through the widespread use of long-lasting insecticide treated bed-nets (LLINs) and artemisinin-based combination therapy, resurgence of malaria is observed in some settings after several years of LLINs use. This study aimed to assess if LLINs use remains protective against malaria during a period of resurgence of malaria morbidity in Dielmo, a rural village of Senegal. Methods: In July 2008, LLINs were offered to all villagers and lately in July 2011, LLINs were renewed.A longitudinal study was conducted between July, 2010 and December, 2011 among inhabitants of the village of Dielmo to identify all episodes of fever. Thick smears stained with Giemsa were done for every febrile villager and malaria attacks were treated with combination of Artesunate plus Amodiaquine. Cross-sectional surveys were also conducted at the end of the rainy season (October 2010 and November 2011) to assess asymptomatic carriage. A survey on LLINs use was done every quarter of the year. A random-effect logistic regression was used to assess the effect of LLINs use on the risk of having a malaria attack after adjusting for the main risk factors. Results: The study population included 449 individuals corresponding to a total of 2140 observations. One hundred and fifteen (115) clinical malaria attacks attributed to P. falciparum (cases) have been recorded over the study period. Most of the malaria cases occurred in October-December 2010 (49/115 i.e. 43%) and among adults aged 15 years and over (50/115, i.e. 43%). During the study period, the use of LLINs was 61% among non-malaria cases and only 42% among malaria clinical cases but differenced according to age group.After adjusting on gender, age, rainfall and LLINs replacement, we found that LLINs use (AOR [95%CI] = 0.40 [0.25; 0.62], p < 0.001) remained a protective factor against malaria attacks during the study period. Conclusion: LLINs use remains effective to reduce malaria burden. These results highlight the need to pursue LLINs implementation in the current context of malaria elimination and to provide positive incentives to increase its use in the population.

Background: Rapid, accurate and high-throughput identification of vector arthropods is of paramount importance in surveillance programmes that are becoming more common due to the changing geographic occurrence and extent of many arthropod-borne diseases. Protein profiling by MALDI-TOF mass spectrometry fulfils these requirements for identification, and reference databases have recently been established for several vector taxa, mostly with specimens from laboratory colonies. Methods: We established and validated a reference database containing 20 phlebotomine sand fly (Diptera: Psychodidae, Phlebotominae) species by using specimens from colonies or field-collections that had been stored for various periods of time. Results: Identical biomarker mass patterns (‘superspectra’) were obtained with colony- or field-derived specimens of the same species. In the validation study, high quality spectra (i.e. more than 30 evaluable masses) were obtained with all fresh insects from colonies, and with 55/59 insects deep-frozen (liquid nitrogen/-80 °C) for up to 25 years. In contrast, only 36/52 specimens stored in ethanol could be identified. This resulted in an overall sensitivity of 87% (140/161); specificity was 100%. Duration of storage impaired data counts in the high mass range, and thus cluster analyses of closely related specimens might reflect their storage conditions rather than phenotypic distinctness. A major drawback of MALDI-TOF MS is the restricted availability of in-house databases and the fact that mass spectrometers from 2 companies (Bruker, Shimadzu) are widely being used. We have analysed fingerprints of phlebotomine sand flies obtained by automatic routine procedure on a Bruker instrument by using our database and the software established on a Shimadzu system. The sensitivity with 312 specimens from 8 sand fly species from laboratory colonies when evaluating only high quality spectra was 98.3%; the specificity was 100%. The corresponding diagnostic values with 55 field-collected specimens from 4 species were 94.7% and 97.4%, respectively. Conclusions: A centralized high-quality database (created by expert taxonomists and experienced users of mass spectrometers) that is easily amenable to customer-oriented identification services is a highly desirable resource. As shown in the present work, spectra obtained from different specimens with different instruments can be analysed using a centralized database, which should be available in the near future via an online platform in a cost-efficient manner.

Background: Triatomine bugs are blood-sucking insects, vectors of Chagas disease. Despite their importance, their oviposition behavior has received relatively little attention. Some triatomines including Rhodnius prolixus stick their eggs to a substrate. It is known that mechanical cues stimulate oviposition in this species. However, it is not clear if chemical signals play a role in this behavior. We studied the role of host cues, including host odor, in the oviposition behavior of the triatomine R. prolixus. Methods: Tests were carried out in an experimental arena and stimuli consisted of a mouse or hen feathers. The number of eggs laid and the position of those eggs with respect to the stimulus source were recorded. Data were analyzed using the Mann-Whitney and Kruskal-Wallis tests. Results: Both a mouse and hen feathers stimulated oviposition. In addition, hen feathers evoked a particular spatial distribution of eggs that was not observed in the case of mouse. Conclusions: We propose that volatile chemical cues from the host play a role in the oviposition behavior of triatomines that stick their eggs. Thus, host odor would stimulate and spatially guide oviposition.

Background: Toxocara canis is a nematode that parasitizes dogs, while humans are paratenic hosts. When humans are infected the migrating larvae damage the liver, lungs and even the nervous system. Larva migrans diagnosis is based on immunological techniques; however, the commercial immunodiagnostic kits detect anti-T. canis antibodies which may cross-react with other parasites, mainly nematodes with extra-intestinal migration. Moreover, antibodies do not necessarily reflect an active infection; so detection and quantification of circulating antigens may provide appropriate and timely information for treatment, which prevents irreversible damage. Here we report the standardization of a monoclonal antibody based antigen capture ELISA to diagnose human toxocariasis without cross-reaction. Methods: We developed anti-T. canis polyclonal antibodies in rabbits and a monoclonal antibody in mouse which did not cross-react with 15 antigens from several parasites. The sandwich ELISA standardization was performed using sera from mice experimentally infected. We tested the method using 29 positive and 58 negative human sera previously typified with a commercial kit, which detects antibodies. Results: Only 5.0 μg/mL and 10 μg/mL polyclonal antibodies and monoclonal antibody, respectively, were needed in the sandwich ELISA standardization, detecting since 440 pg/mL larva antigens. Nine out of 29 antibody-positive sera were also positive for antigens and no false positive were found. Taking the antibody kit as the reference standard, the sensibility and specificity of the antigen test were 31% and 100%, respectively. Conclusions: With these tools we established a detection threshold as low as 440 pg/mL antigen. Monoclonal antibody is specific, and did not cross-react with antigens from other parasites. Detection of circulating antigens helps provide appropriate and timely treatment and prevents irreversible damage.

Background: In general, there was a large reduction in the occurrence of cases of Chagas disease in the last decades in Brazil. However, despite all of these efforts, there have been various reports of persistent reinfestations of T. sordida in a large part of the state of Minas Gerais, for reasons still little investigated. Thus, this purpose of this study was to characterize the deltamethrin susceptibility profile of peridomestic T. sordida populations from North of Minas Gerais – Brazil. Methods: Susceptibility to deltamethrin was assessed in seventeen peridomestic populations of T. sordida from North region of Minas Gerais, Brazil. Serial dilutions of deltamethrin in acetone (0.2 μL) were topically applied in first instar nymphs (F1, five days old, fasting, weight 1.2 ± 0.2 mg). Dose response results were analyzed with POLO program, determining the lethal doses, slope and resistance ratios (RR). Results: Susceptibility profile characterization of T. sordida populations revealed resistance ratios (RR50) ranging from 2.50 to 7.27. Conclusions: In fact, we know very little about the real impact of the resistance ratios obtained in the laboratory bioassays on the effectiveness of the vector control activities in the field. Thus, we prefer to refer to the populations with RR > 5 as populations with altered susceptibility. For these populations, the realization of laboratory and field trials, simultaneous and complementary, permitting the evaluation of both, is recommended.

Background: Newly recognized endemic foci for human babesiosis include Europe, where Ixodes ricinus, a vector for several species of Babesia, is the most commonly identified tick. Vector-based surveillance provides an early warning system for the emergence of human babesiosis, which is likely to be under-reported at emerging sites. In the present study, we set out to screen I. ricinus collected from Danish domestic dogs for Babesia, in order to identify whether humans in Denmark are exposed to the parasite.FindingsA total of 661 ticks (Ixodes spp.) were collected from 345 Danish domestic dogs during April-September 2011 and pooled, one sample per dog. DNA was extracted from each sample and examined by PCR and sequencing for Rickettsia spp., Borrelia burgdorferi sensu lato, Bartonella spp., Francisella tularensis, Candidatus Neoehrlichia mikurensis, and Babesia spp. In total, 34% of the samples were positive for tick-borne microorganisms potentially pathogenic to humans: Rickettsia spp. were detected in 16% of the pools, with 79% being R. helvetica. Borrelia burgdorferi sensu lato was found in 15%, with the main species identified as Borrelia afzelii (39%). Likewise, 8% of the samples were positive for Babesia spp. (Babesia microti, 82%; Babesia venatorum (‘EU1’), 18%). Lastly, 1% of the samples tested positive for Candidatus Neoehrlichia mikurensis, and 0.6% for Bartonella spp. No ticks were found to be infected with Francisella tularensis. Conclusions: Our data are in support of endemic occurrence of potentially zoonotic Babesia in Denmark and confirms I. ricinus as a vector of multiple pathogens of public health concern.

Background: Focal screening and treatment (FSAT) of malaria infections has recently been introduced in Peru to overcome the inherent limitations of passive case detection (PCD) and further decrease the malaria burden. Here, we used a relatively straightforward mathematical model to assess the potential of FSAT as elimination strategy for Plasmodium falciparum malaria in the Peruvian Amazon Region. Methods Methods: A baseline model was developed to simulate a scenario with seasonal malaria transmission and the effect of PCD and treatment of symptomatic infections on the P. falciparum malaria transmission in a low endemic area of the Peruvian Amazon. The model was then adjusted to simulate intervention scenarios for predicting the long term additional impact of FSAT on P. falciparum malaria prevalence and incidence. Model parameterization was done using data from a cohort study in a rural Amazonian community as well as published transmission parameters from previous studies in similar areas. The effect of FSAT timing and frequency, using either microscopy or a supposed field PCR, was assessed on both predicted incidence and prevalence rates. Results: The intervention model indicated that the addition of FSAT to PCD significantly reduced the predicted P. falciparum incidence and prevalence. The strongest reduction was observed when three consecutive FSAT were implemented at the beginning of the low transmission season, and if malaria diagnosis was done with PCR. Repeated interventions for consecutive years (10 years with microscopy or 5 years with PCR), would allow reaching near to zero incidence and prevalence rates. Conclusions Conclusions: The addition of FSAT interventions to PCD may enable to reach P. falciparum elimination levels in low endemic areas of the Amazon Region, yet the progression rates to those levels may vary substantially according to the operational criteria used for the intervention.

Background: The pathogenesis of S. mansoni infection involves chronic inflammatory responses to parasite eggs which can be associated with a characteristic periportal fibrosis (PPF) and the progression to severe hepatosplenic disease. The effects of HIV-1 co-infection and the influence of CD4+ cell numbers on these clinical manifestations of chronic S. mansoni are not known. To understand the effects of HIV-1 co-infection on these morbidities, we examined S. mansoni ultrasound-detectable morbidities in relation to HIV-1 infection and CD4+ cell counts, and other factors in fishing communities where the two infections are present. Methods: Ultrasonographical examination was conducted during a cross-sectional study of 1,671 (aged 21–55 years) individuals in North-Western Tanzania. Blood samples were obtained for HIV-1 screening and CD4+ cell quantification. A single stool sample was examined for S. mansoni eggs using the Kato-Katz technique. A questionnaire was used to collect socio-demographic-economic information. Results: The prevalence of PPF (grade C-F) was 13.79% and 15.01% for the HIV-1 infected and non-infected individuals (P = 0.72). Male gender (P< 0.001), age group 21–30 years (P< 0.028) and, residential time of 11–20 (P< 0.01) and ≥21 years (P< 0.01) were associated with PPF in S. mansoni infected individuals. The height-adjusted measurements of the left liver lobe were significantly larger in HIV-1/S. mansoni co-infected compared to S. mansoni only-infected individuals (t = −2.0702, P< 0.039). Predictors of the height-adjusted measurements of the left liver lobe and spleen were age, male gender, malaria infection, fishing occupation, village of residence and heavy intensity of S. mansoni infection. After accounting for these factors, neither HIV-1 infection nor CD4+ cell counts predicted PPF, hepatosplenomegaly, measurements of the liver or spleen. Height-adjusted ultrasound measurements of the left liver lobe did not correlate with the CD4+ cells counts in co-infected individuals (r = −0.16, P = 0.084). Conclusion: S. mansoni-related PPF, liver and spleen enlargement are prevalent in the study population. The intensity of S. mansoni infection was associated with the enlargement of liver, spleen and hepatosplenomegaly. The PPF grades observed were similar in both HIV-1/S. mansoni co-infected and in those only infected with S. mansoni. There was no evidence that HIV-1 infection or CD4+ cells counts were associated with these S. mansoni morbidities.

Background: Triatoma infestans, the main vector of Trypanosoma cruzi (causative agent of Chagas disease) has been successfully eliminated over much of its original geographic distribution over the southern cone countries of South America. However, populations of the species are still infesting houses of rural communities of the Gran Chaco region of Argentina and Bolivia. This study reports for the first time a large-scale effect of a vector control intervention using a microencapsulated formulation of organophosphates and insect growth regulator on house infestation by T. infestans, in the southwestern region of Santa Cruz de la Sierra Department, within the Bolivian chaco. Methods: The vector control intervention included the treatment and entomological evaluation of 1626 individually coded and georeferenced houses with the microencapsulated formulation. House infestation by T. infestans was evaluated by active searches with fixed capture effort carried out before and after two, 16 and 32 months of the treatment application. Results: House infestation prevalence was 30.5% before the intervention, spatially aggregated in two clusters of 38 and 25 localities that showed 41% and 38% house infestation by T. infestans. Infestation prevalence was reduced to 2.4% two months after the intervention and remained very low (1.7%) until the end of the study after 32 months of the control intervention, without any other additional vector control intervention. Conclusions: The obtained results show an important long lasting effect on house protection against triatomine infestation in a region of known pyrethroid resistant populations of T. infestans, as the result of the slow release of the active ingredients, protected by the formulation microcapsule.

Background: The application of the sterile insect technique (SIT) requires mass-production of sterile males of good biological quality. The size of the project area will in most cases determine whether it is more cost effective to produce the sterile flies locally (and invest in a mass-rearing facility) or import the sterile flies from a mass-rearing facility that is located in another country. This study aimed at assessing the effect of long distance transport of sterile male Glossina palpalis gambiensis pupae on adult male fly yield. Methods: The male pupae were produced at the Centre International de Recherche-Développement sur l’Elevage en zone Subhumide (CIRDES), Bobo-Dioulasso, Burkina Faso, and shipped with a commercial courier service in insulated transport boxes at a temperature of ±10°C to Senegal (±36 h of transport). Upon arrival in the insectary in Dakar, the pupae were transferred to an emergence room and the flies monitored for 3–6 days. Results: The results showed that the used system of isothermal boxes that contained phase change material packs (S8) managed to keep the temperature at around 10°C which prevented male fly emergence during transport. The emergence rate was significantly higher for pupae from batch 2 (chilled at 4°C for one day in the source insectary before transport) than those from batch 1 (chilled at 4°C for two days in the source insectary before transport) i.e. an average (±sd) of 76.1 ± 13.2% and 72.2 ± 14.3%, respectively with a small proportion emerging during transport (0.7 ± 1.7% and 0.9 ± 2.9%, respectively). Among the emerged flies, the percentage with deformed (not fully expanded) wings was significantly higher for flies from batch 1 (12.0 ± 6.3%) than from batch 2 (10.7 ± 7.5%). The amount of sterile males available for release as a proportion of the total pupae shipped was 65.8 ± 13.3% and 61.7 ± 14.7% for batch 1 and 2 pupae, respectively. Conclusions: The results also showed that the temperature inside the parcel must be controlled around 10°C with a maximal deviation of 3°C to maximize the male yield.

Background: Landscape modifications, urbanization or changes of use of rural-agricultural areas can create more favourable conditions for certain mosquito species and therefore indirectly cause nuisance problems for humans. This could potentially result in mosquito-borne disease outbreaks when the nuisance is caused by mosquito species that can transmit pathogens. Anopheles plumbeus is a nuisance mosquito species and a potential malaria vector. It is one of the most frequently observed species in the Netherlands. Information on the distribution of this species is essential for risk assessments. The purpose of the study was to investigate the potential spatial distribution of An. plumbeus in the Netherlands. Methods: Random forest models were used to link the occurrence and the abundance of An. plumbeus with environmental features and to produce distribution maps in the Netherlands. Mosquito data were collected using a cross-sectional study design in the Netherlands, from April to October 2010–2013. The environmental data were obtained from satellite imagery and weather stations. Statistical measures (accuracy for the occurrence model and mean squared error for the abundance model) were used to evaluate the models performance. The models were externally validated. Results: The maps show that forested areas (centre of the Netherlands) and the east of the country were predicted as suitable for An. plumbeus. In particular high suitability and high abundance was predicted in the south-eastern provinces Limburg and North Brabant. Elevation, precipitation, day and night temperature and vegetation indices were important predictors for calculating the probability of occurrence for An. plumbeus. The probability of occurrence, vegetation indices and precipitation were important for predicting its abundance. The AUC value was 0.73 and the error in the validation was 0.29; the mean squared error value was 0.12. Conclusions: The areas identified by the model as suitable and with high abundance of An. plumbeus, are consistent with the areas from which nuisance was reported. Our results can be helpful in the assessment of vector-borne disease risk.

Background: Ixodes holocyclus ticks are a frequently fatal threat to dogs in eastern Australia. These ticks secrete a neurotoxin that can produce an ascending paralysis after 72 h attachment that can lead to death in affected animals. Fluralaner is a potent systemic acaricide with immediate and persistent efficacy for tick control including evidence of 100% efficacy against Ixodes ricinus ticks within 72 h. This study investigated the potential for oral fluralaner administration to control I. holocyclus infestation and the subsequent risk of host paralysis. Methods: Healthy Foxhound and Foxhound cross dogs immunized against holocyclotoxin were randomly allocated to receive either a single fluralaner (at least 25 mg/kg) dose or no treatment. All dogs were penned individually and infested with 30 adult unfed female I. holocyclus 1 day before treatment and 14, 28, 42, 56, 70, 84, 112 and 140 days following treatment. Ticks were counted and assessed at 24, 48 and 72 h after the initial fluralaner treatment and after each subsequent infestation. Ticks were not removed at the 24 and 48 h assessments, but were removed after the 72 h assessments. On 112 and 140 days post treatment a new group of untreated control dogs was used. Results: Fluralaner treatment efficacy against I. holocyclus was 100% at 72 h post treatment. Following re-infestations the efficacy remained at 100% at the 72 h assessments for 115 days and reached 95.7% at 143 days. The differences between mean live tick counts on treatment and control groups were significant (P < 0.00l) at all assessment time points for 143 days following treatment. Conclusions: Oral fluralaner treatment can prevent Australian paralysis tick infestations for at least 115 days.