Parasites & Vectors

Publications in this journal

• Article: Altitudinal variation in haemosporidian parasite distribution in great tit populations.

  Juan van Rooyen, Fabrice Lalubin, Olivier Glaizot, Philippe Christe
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  ABSTRACT: BACKGROUND: One of the major issues concerning disease ecology and conservation is knowledge of the factors that influence the distribution of parasites and consequently disease outbreaks. This study aimed to investigate avian haemosporidian composition and the distribution of these parasites in three altitudinally separated great tit (Parus major) populations in western Switzerland over a three-year period. The objectives were to determine the lineage diversity of parasites occuring across the study populations and to investigate whether altitudinal gradients govern the distribution of haemosporidian parasites by lineage. METHODS: In this study molecular approaches (PCR and sequencing) were used to detect avian blood parasites (Plasmodium sp., Haemoproteus sp. and Leucocytozoon sp.) in populations of adult great tits caught on their nests during three consecutive breeding seasons. RESULTS: High levels of parasite prevalence (88-96%) were found across all of the study populations with no significant altitude effect. Altitude did, however, govern the distribution of parasites belonging to different genera, with Plasmodium parasites being more prevalent at lower altitudes, Leucocytozoon parasites more at high altitude and Haemoproteus parasite prevalence increasing with altitude. A total of 27 haemosporidian parasite lineages were recorded across all study sites, with diversity showing a positive correlation to altitude. Parasites belonging to lineage SGS1 (P. relictum) and PARUS4 and PARUS19 (Leucocytozoon sp.) dominated lower altitudes. SW2 (P. polare) was the second most prevalent lineage of parasite detected overall and these parasites were responsible for 68% of infections at intermediate altitude, but were only documented at this one study site. CONCLUSIONS: Avian haemosporidian parasites are not homogeneously distributed across host populations, but differ by altitude. This difference is most probably brought about by environmental factors influencing vector prevalence and distribution. The high occurrence of co-infection by different genera of parasites might have pronounced effects on host fitness and should consequently be investigated more rigorously.
  Parasites & Vectors 05/2013; 6(1):139.
• Article: Spatio-temporal analysis to identify determinants of Oncomelania hupensis infection with Schistosoma japonicum in Jiangsu province, China.

  Kun Yang, Wei Li, Le-Ping Sun, Yi-Xin Huang, Jian-Feng Zhang, Feng Wu, De-Rong Hang, Peter Steinmann, You-Sheng Liang
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  ABSTRACT: BACKGROUND: With the successful implementation of integrated measures for schistosomiasis control, Jiangsu province has reached low-endemicity status. However, infected Oncomelania hupensis snails could still be found in certain locations along the Yangtze river until 2009, and there is concern that they might spread again, resulting in the possible re-emergence of infections among people and domestic animals alike. In order to establish a robust surveillance system that is able to detect the spread of infected snails at an early stage, sensitive and reliable methods to identify risk factors for the establishment of infected snails need to be developed. METHODS: A total of 107 villages reporting the persistent presence of infected snails were selected. Relevant data on the distribution of infected snails, and human and livestock infection status information for the years 2003 to 2008 were collected. Spatio-temporal pattern analysis including spatial autocorrelation, directional distribution and spatial error models were carried out to explore spatial correlations between infected snails and selected explanatory factors. RESULTS: The area where infected snails were found, as well as their density, decreased significantly between 2003 and 2008. Changes in human and livestock prevalences were less pronounced. Three statistically significant spatial autocorrelations for infected snails were identified. (i) The Moran's I of infected snails increased from 2004 to 2007, with the snail density increasing and the area with infected snails decreasing. (ii) The standard deviations of ellipses around infected snails were decreasing and the central points of the ellipses moved from West to East. (iii) The spatial error models indicated no significant correlation between the density of infected snails and risk factors. CONCLUSIONS: We conclude that the contribution of local infection sources including humans and livestock to the distribution of infected snails might be relatively small and that snail control may limit infected snails to increasingly small areas ecologically most suitable for transmission. We provide a method to identify these areas and risk factors for persistent infected snail presence through spatio-temporal analysis, and a suggested framework, which could assist in designing evidence based control strategies for schistosomiasis elimination.
  Parasites & Vectors 05/2013; 6(1):138.
• Article: Malaria infection does not affect the sensitivity of peripheral receptor neurons in Anopheles stephensi.

  Alan J Grant, Marc A Muskavitch, Robert J O Connell
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  ABSTRACT: BACKGROUND: Mosquitoes transmit many important diseases including malaria, dengue and yellow fever. Disease transmission from one vertebrate host to another depends on repeated blood feedings by single mosquitoes. In order for the mosquito to acquire the blood that it needs to complete oogenesis, the insect must locate a suitable host. Olfactory cues (including carbon dioxide) released by the host and detected by the mosquito are the primary signals that vector insects use for host location. Previous studies have suggested that the physiological status - including bacterial, fungal, viral and Plasmodium infections - can modulate aspects of behavior in haematophagous insects. METHODS: Standard electrophysiological techniques were used to record extracellular responses from the receptor neurons located in sensilla found on the maxillary palps of the insects. The recording microelectrode was inserted through the cuticle at the base of an individual sensillum and the extracellular electrical signals obtained from the three neurons within the sensillum were recorded. Stimulations consisted of 2 s pulses of the desired concentrations of CO2 or dosages of 1-octen-3-ol. RESULTS: Accordingly, we were interested in determining whether Plasmodium infection affects the sensitivity of those peripheral olfactory sensors that are involved in host-seeking in mosquitoes. Our studies indicate that infection of female Anopheles stephensi with Plasmodium berghei does not alter the response characteristics of the neurons innervating the maxillary palp sensilla that respond to the attractants carbon dioxide and 1-octen-3-ol. Although the response characteristics of the peripheral sensory neurons are not affected by infection status, we found that the age of the mosquito alone does affect the threshold of sensitivity of these neurons to carbon dioxide. The proportion of older insects (21--30 d post-emergence) that responds to 150 ppm carbon dioxide is higher than the proportion that responds among younger insects (1--10 d post-emergence). CONCLUSIONS: Anopheles stephensi infected with Plasmodium berghei exhibit sensitivities to stimulation with carbon dioxide and 1-octen-3-ol similar to those of uninfected mosquitoes. However, the age of the infected or uninfected mosquito does affect the threshold of sensitivity of these neurons to carbon dioxide.
  Parasites & Vectors 05/2013; 6(1):134.
• Article: Anopheles species associations in Southeast Asia: indicator species and environmental influences.

  Valérie Obsomer, Marc Dufrene, Pierre Defourny, Marc Coosemans
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  ABSTRACT: BACKGROUND: Southeast Asia presents a high diversity of Anopheles. Environmental requirements differ for each species and should be clarified because of their influence on malaria transmission potential. Monitoring projects collect vast quantities of entomological data over the whole region and could bring valuable information to malaria control staff but collections are not always standardized and are thus difficult to analyze. In this context studying species associations and their relation to the environment offer some opportunities as they are less subject to sampling error than individual species. METHODS: Using asymmetrical similarity coefficients, indirect clustering and the search of indicator species, this paper identified species associations. Environmental influences were then analysed through canonical and discriminant analysis using climatic and topographic data, land cover in a 3 km buffer around villages and vegetation indices. RESULTS: Six groups of sites characterized the structure of the species assemblage. Temperature, rainfall and vegetation factors all play a role. Four out of the six groups of sites based on species similarities could be discriminated using environmental information only. CONCLUSIONS: Vegetation indices derived from satellite imagery proved very valuable with one variable explaining more variance of the species dataset than any other variable. The analysis could be improved by integrating seasonality in the sampling and collecting at least 4 consecutive days.
  Parasites & Vectors 05/2013; 6(1):136.
• Article: Hepatic stellate cells increase in Toxoplasma gondii infection in mice

  Atmaca HT, Gazyagci AN, Canpolat S, Kul O
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  ABSTRACT: Background Toxoplasma gondii is a ubiquitous protozoan parasite that can infect humans and animals. The severity of toxoplasmosis varies according to the immune status of the individual, parasite strain, and host species. In mammalian species, it has been observed that severe lesions of acute toxoplasmosis form in visceral organs such as the liver, lung, and spleen. Some epidemiological studies have reported an association of T. gondii infection with liver cirrhosis. Methods Acute infection was induced in fifteen 30-day-old normal Swiss albino mice. The mice were infected by intraperitoneal inoculation of 5000 T. gondii RH strain tachyzoites. The mice were sacrificed in groups of 5 at 2, 4, and 6 days after inoculation. Another group of 5 mice were used as the controls. Anti-glial fibrillary acidic protein (GFAP) and anti-T. gondii antibodies were used to compare GFAP-immunoreactive cells and anti-T. gondii--immunopositive areas in the liver between the T. gondii-infected groups and the healthy controls, respectively. Results There was a significant correlation between the numbers of GFAP-positive hepatic stellate cells (HSCs) when they were compared with T. gondii antigen immunostaining (p < 0.05). The amount of T. gondii immunostaining increased significantly with the increase in the number of HSCs. Conclusions There is a significant relationship between the number of HSCs and T. gondii antigens, which may represent an active role of HSCs in liver pathology and the pathobiology of T. gondii-related hepatitis.
  Parasites & Vectors 05/2013; 6(135).
• Article: Insecticide-impregnated netting as a potential tool for long-lasting control of the leishmaniasis vector Lutzomyia longipalpis in animal shelters.

  Daniel Peter Bray, James G Hamilton
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  ABSTRACT: BACKGROUND: Leishmaniasis remains a serious neglected disease, with more than 350 million people potentially at risk worldwide. Control strategies often rely on spraying residual insecticides to target populations of the sand fly vectors that transmit Leishmania parasites when blood-feeding. These programmes are often difficult to sustain effectively, as sand fly resting sites must be resprayed on a regular basis. Here, we investigate whether application of insecticide-impregnated netting to a surface could act as an alternative to residual spraying for controlling the American visceral leishmaniasis vector Lutzomyia longipalpis. METHODS: Female L. longipalpis from our laboratory colony were exposed for 1 h to three treatments applied to plywood surfaces: 2% permethrin-impregnated netting (Olyset(R)), 20 mg a.i.m-2 micro-encapsulated lambda-cyhalothrin (Demand CS(R)) and a no-treatment control. We compared the speed at which these treatments acted, by measuring the percentage of sand flies killed both immediately after exposure to the treatment for 1 hour, as well as the number that had died 24 h after the 1 hour exposure. We repeated the experiment at 6 and 12 months following application to test the effectiveness of each treatment over time. RESULTS: When first applied, the lambda-cyhalothrin killed more sand flies in the first hour than the permethrin-impregnated netting. However, the effectiveness of the lambda-cyhalothrin diminished over time, so that there was no difference between the two treatments at 12 months. Both killed more sand flies than the control. When measured 24 h following exposure, both test treatments had killed close to 100% of sand flies when first applied, but while the lethal effect of the netting was maintained at close to 100% over 12 months, the effectiveness of the residual insecticide diminished to approximately 80% after 6 months. CONCLUSIONS: The results of these initial laboratory experiments indicate that covering surfaces with insecticide impregnated netting material may provide a longer-lasting solution for killing sand flies than residual spraying. Field trials are needed to identify the feasibility of treating surfaces with netting or similar impregnated materials as part of a control program. In targeting L. longipalpis, the greatest benefits may be seen in treating animal sheds with netting, where these sand flies aggregate in large numbers, and which can be difficult to treat repeatedly by conventional spraying.
  Parasites & Vectors 05/2013; 6(1):133.
• Article: Redescription of Cercopithifilaria bainae Almeida & Vicente, 1984 (Spirurida, Onchocercidae) from a dog in Sardinia, Italy.

  Domenico Otranto, Antonio Varcasia, Cinzia Solinas, Antonio Scala, Emanuele Brianti, Filipe Dantas-Torres, Giada Annoscia, Coralie Martin, Yasen Mutafchiev, Odile Bain
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  ABSTRACT: BACKGROUND: Three species of the genus Cercopithifilaria have been morphologically and molecularly characterized in dog populations in southern Europe: Cercopithifilaria grassii (Noe, 1907), Cercopithifilaria sp. sensu Otranto et al., 2011 (reported as Cercopithifilaria sp. I), and Cercopithifilaria sp. II sensu Otranto et al., 2012. The adults of Cercopithifilaria sp. I have remained unknown until the present study. METHODS: The material originated from a dog from Sardinia (Italy) diagnosed with dermal microfilariae of Cercopithifilaria sp. I. The holotype and three paratypes of Cercopithifilaria bainae Almeida & Vicente, 1984, described from dogs in Brazil, were studied as comparative material. A cox1 (~689 bp) and 12S (~330 bp) gene fragments were amplified and phylogenetic analysis carried out. RESULTS: The highest numbers of adult nematodes (82%) were collected in the sediment of the subcutaneous tissues of the trunk (n = 37) and forelimbs (n = 36). The morphology of the adult nematodes and microfilariae collected from the dog in Sardinia corresponded to those of C. bainae. All cox1 and 12S gene sequences showed a high homology (99-100%) with sequences from microfilariae of Cercopithifilaria sp. I. CONCLUSIONS: The morphological and molecular identity of the microfilariae of C. bainae overlap those described previously as Cercopithifilaria sp. sensu Otranto et al., 2011 (=Cercopithifilaria sp. I). Therefore, the present study reports the occurrence of C. bainae in Europe, for the first time after its description and the single record in Brazil. C. bainae appears to be highly diffused in dog populations in southern Europe. The phylogenetic analyses based on cox1 and 12S do not reveal the three species of Cercopithifilaria parasitizing dogs as a monophyletic group, which suggests that they have derived independently by host switching.
  Parasites & Vectors 05/2013; 6(1):132.
• Article: Hepatic stellate cells increase in Toxoplasma gondii infection in mice.

  Hasan Tar K Atmaca, Aycan Nuriye Gazya C, S la Canpolat, O Uz Kul
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  ABSTRACT: BACKGROUND: Toxoplasma gondii is a ubiquitous protozoan parasite that can infect humans and animals. The severity of toxoplasmosis varies according to the immune status of the individual, parasite strain, and host species. In mammalian species, it has been observed that severe lesions of acute toxoplasmosis form in visceral organs such as the liver, lung, and spleen. Some epidemiological studies have reported an association of T. gondii infection with liver cirrhosis. METHODS: Acute infection was induced in fifteen 30-day-old normal Swiss albino mice. The mice were infected by intraperitoneal inoculation of 5000 T. gondii RH strain tachyzoites. The mice were sacrificed in groups of 5 at 2, 4, and 6 days after inoculation. Another group of 5 mice were used as the controls. Anti-glial fibrillary acidic protein (GFAP) and anti-T. gondii antibodies were used to compare GFAP-immunoreactive cells and anti-T. gondii--immunopositive areas in the liver between the T. gondii-infected groups and the healthy controls, respectively. RESULTS: There was a significant correlation between the numbers of GFAP-positive hepatic stellate cells (HSCs) when they were compared with T. gondii antigen immunostaining (p < 0.05). The amount of T. gondii immunostaining increased significantly with the increase in the number of HSCs. CONCLUSIONS: There is a significant relationship between the number of HSCs and T. gondii antigens, which may represent an active role of HSCs in liver pathology and the pathobiology of T. gondii-related hepatitis.
  Parasites & Vectors 05/2013; 6(1):135.
• Article: Taxis assays measure directional movement of mosquitoes to olfactory cues.

  Lena M Lorenz, Aidan Keane, Jason D Moore, Cristina J Munk, Laura Seeholzer, Antony Mseka, Emmanuel Simfukwe, Joseph Ligamba, Elizabeth L Turner, Lubandwa R Biswaro, Fredros O Okumu, Gerry F Killeen, Wolfgang R Mukabana, Sarah J Moore
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  ABSTRACT: BACKGROUND: Malaria control methods targeting indoor-biting mosquitoes have limited impact on vectors that feed and rest outdoors. Exploiting mosquito olfactory behaviour to reduce blood-feeding outdoors might be a sustainable approach to complement existing control strategies. Methodologies that can objectively quantify responses to odour under realistic field conditions and allow high-throughput screening of many compounds are required for development of effective odour-based control strategies. METHODS: The olfactory responses of laboratory-reared Anopheles gambiae in a semi-field tunnel and A. arabiensis females in an outdoor field setting to three stimuli, namely whole human odour, a synthetic blend of carboxylic acids plus carbon dioxide and CO2 alone at four distances up to 100 metres were measured in two experiments using three-chambered taxis boxes that allow mosquito responses to natural or experimentally-introduced odour cues to be quantified. RESULTS: Taxis box assays could detect both activation of flight and directional mosquito movement. Significantly more (6-18%) A. arabiensis mosquitoes were attracted to natural human odour in the field up to 30 metres compared to controls, and blended synthetic human odours attracted 20% more A. gambiae in the semi-field tunnel up to 70 metres. Whereas CO2 elicited no response in A. arabiensis in the open field, it was attractive to A. gambiae up to 50 metres (65% attraction compared to 36% in controls). CONCLUSIONS: We have developed a simple reproducible system to allow for the comparison of compounds that are active over medium- to long-ranges in semi-field or full-field environments. Knowing the natural range of attraction of anopheline mosquitoes to potential blood sources has substantial implications for the design of malaria control strategies, and adds to the understanding of olfactory behaviour in mosquitoes. This experimental strategy could also be extended from malaria vectors to other motile arthropods of medical, veterinary and agricultural significance.
  Parasites & Vectors 05/2013; 6(1):131.
• Article: Efficacy of indoxacarb applied to cats against the adult cat flea, Ctenocephalides felis, flea eggs and adult flea emergence.

  Michael W Dryden, Patricia A Payne, Vicki Smith, Kathleen Heaney, Fangshi Sun
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  ABSTRACT: BACKGROUND: A study was conducted to evaluate the effect of indoxacarb applied to cats on adult cat fleas, Ctenocephalides felis, flea egg production and adult flea emergence. METHODS: Sixteen cats were selected for the study and allocated to two treatment groups. Eight cats were treated with a 19.5% w/v topical spot-on solution of indoxacarb on day 0 and eight cats served as untreated controls. Each cat was infested with 50 fleas on Days -2, 7, 14, 21, 28, 35 and 42. On Days 1, 2, and 3, and at 2 and 3 days after each post treatment reinfestation flea eggs were collected from the pan under each cat cage. Eggs were counted and viability assessed by evaluating adult flea emergence 28 days after egg collection. Three days after treatment or infestation, each cat was combed to remove and count live fleas. RESULTS: Treatment with indoxacarb provided 100% efficacy following infestations on day -2, 7, 14, 21 and 28 and efficacy was 99.6% following infestations on days 35 and 42. Egg production from indoxacarb treated cats was reduced by 99.9% within 72 hours of treatment. For subsequent infestations no eggs were produced from treated cats from day 8 through day 30. Egg production was still reduced by >=95.8% through day 45. Indoxacarb treatment also reduced adult flea emergence from eggs for 5 weeks after treatment. The combination of reduction in egg numbers and egg viability from indoxacarb treated cats reduced predicted flea emergence by 100% from days 2 -- 31 and 99.9%, 100%, 96.4% and 99.0% on days 37, 38, 44 and 45, respectively. CONCLUSIONS: A topical spot-on formulation of indoxacarb provided >=99.6% efficacy against flea infestations on cats for 6 weeks following a single treatment. Indoxacarb also eliminated or markedly reduced egg production for the entire evaluation period and reduced the viability of the few eggs that were produced from Day 1 through Day 38. Given indoxacarb's effect on adult fleas, egg production and egg viability; this formulation can interrupt flea reproduction on treated cats for at least 6 weeks after treatment.
  Parasites & Vectors 05/2013; 6(1):126.
• Article: Bioefficacy of long-lasting insecticidal nets against pyrethroid-resistant populations of Anopheles gambiae s.s. from different malaria transmission zones in Uganda.

  Michael Okia, Richard Ndyomugyenyi, James Kirunda, Anatol Byaruhanga, Seraphine Adibaku, Denis K Lwamafa, Fred Kironde
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  ABSTRACT: BACKGROUND: There are major concerns over sustaining the efficacy of current malaria vector control interventions given the rapid spread of resistance, particularly to pyrethroids. This study assessed the bioefficacy of five WHO-recommended long-lasting insecticidal nets (LLINs) against pyrethroid-resistant Anopheles gambiae field populations from Uganda. METHODS: Adult An. gambiae from Lira, Tororo, Wakiso and Kanungu districts were exposed to permethrin (0.75%) or deltamethrin (0.05%) in standard WHO susceptibility tests. Cone bioassays were used to measure the bioefficacy of four mono-treated LLINs (Olyset(R), Interceptor(R), Netprotect(R) and PermaNet(R) 2.0) and one combination LLIN (PermaNet(R) 3.0) against the four mosquito populations. Wireball assays were similarly conducted to determine knockdown rates. Species composition and kdr mutation frequency were determined for a sample of mosquitoes from each population. Chemical assays confirmed that test nets fell within target dose ranges. RESULTS: Anopheles gambiae s.s. predominated at all four sites (86 - 99% of Anopheles spp.) with moderate kdr L1014S allelic frequency (0.34 -- 0.37). Confirmed or possible resistance to both permethrin and deltamethrin was identified for all four test populations. Reduced susceptibility to standard LLINs was observed for all four populations, with mortality rates as low as 45.8% even though the nets were unused. The combination LLIN PermaNet(R)3.0 showed the highest overall bioefficacy against all four An. gambiae s.l. populations (98.5 - 100% mortality). Wireball assays provided a more sensitive indicator of comparative bioefficacy, and PermaNet 3.0 was again associated with the highest bioefficacy against all four populations (76.5 -- 91.7% mortality after 30 mins). CONCLUSIONS: The bioefficacy of mono-treated LLINs against pyrethroid-resistant field populations of An. gambiae varied by LLIN type and mosquito population, indicating that certain LLINs may be more suitable than others at particular sites. In contrast, the combination LLIN PermaNet 3.0 performed optimally against the four An. gambiae populations tested. The observed reduced susceptibility of malaria vectors to mono-treated LLINs is of particular concern, especially considering all nets were unused. With ongoing scale-up of insecticidal tools in the advent of increasing resistance, it is essential that those interventions with proven enhanced efficacy are given preference particularly in areas with high resistance.
  Parasites & Vectors 05/2013; 6(1):130.
• Article: Predicting Culex pipiens/restuans population dynamics by interval lagged weather data.

  Karin Lebl, Katharina Brugger, Franz Rubel
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  ABSTRACT: BACKGROUND: Culex pipiens/restuans mosquitoes are important vectors for a variety of arthropod borne viral infections. In this study, the associations between 20 years of mosquito capture data and the time lagged environmental quantities daytime length, temperature, precipitation, relative humidity and wind speed were used to generate a predictive model for the population dynamics of this vector species. METHODS: Mosquito population in the study area was represented by averaged time series of mosquitos counts captured at 6 sites in Cook County (Illinois, USA). Cross-correlation maps (CCMs) were compiled to investigate the association between mosquito abundances and environmental quantities. The results obtained from the CCMs were incorporated into a Poisson regression to generate a predictive model. To optimize the predictive model the time lags obtained from the CCMs were adjusted using a genetic algorithm. RESULTS: CCMs for weekly data showed a highly positive correlation of mosquito abundances with daytime length 4 to 5 weeks prior to capture (quantified by a Spearman rank order correlation of rS = 0.898) and with temperature during 2 weeks prior to capture (rS = 0.870). Maximal negative correlations were found for wind speed averaged over 3 week prior to capture (rS = -0.621). Cx. pipiens/restuans population dynamics was predicted by integrating the CCM results in Poisson regression models. They were used to simulate the average seasonal cycle of the mosquito abundance. Verification with observations resulted in a correlation of rS = 0.899 for daily and rS = 0.917 for weekly data. Applying the optimized models to the entire 20-years time series also resulted in a suitable fit with rS = 0.876 for daily and rS = 0.899 for weekly data. CONCLUSIONS: The study demonstrates the application of interval lagged weather data to predict mosquito abundances with a feasible accuracy, especially when related to weekly Cx. pipiens/restuans populations.
  Parasites & Vectors 05/2013; 6(1):129.
• Article: Babesia lengau associated with cerebral and haemolytic babesiosis in two domestic cats.

  Anna-Mari Bosman, Marinda C Oosthuizen, Estelle H Venter, Johan Ca Steyl, Tertius A Gous, Barend L Penzhorn
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  ABSTRACT: BACKGROUND: Although reported sporadically from various countries, feline babesiosis appears to be a significant clinical entity only in South Africa, where Babesia felis is usually incriminated as the causative agent. Babesia lengau, recently described from asymptomatic cheetahs, has now possibly been incriminated as the causative agent in two severe clinical cases in domestic cats. FINDINGS: Both cats were euthanised in extremis. While typical feline babesiosis in South Africa is an afebrile disease with a chronic manifestation, there was acute onset of severe clinical signs in both cats and their body temperatures were above the normal range when they were presented for treatment. Haemolytic anaemia was confirmed in one case. To our knowledge, this is the first report of cerebral babesiosis in cats.On reverse line blot 18S rDNA PCR products obtained from both cats showed positive hybridization profiles with the B. lengau species-specific probe. The two partial parasite 18S rRNA gene sequences obtained, showed high sequence similarity (99.9 %) to B. lengau. In a representative tree constructed by the neighbor-joining method using the two-parameter model of Kimura the two obtained partial 18S rDNA sequences and that of B. lengau formed a monophyletic group with B. conradae and sequences previously isolated from humans and wildlife in the western USA. CONCLUSION: All clinical cases of feline babesiosis in South Africa are not necessarily caused by B. felis. Other piroplasms, e.g. B. lengau, may be incriminated in clinical cases, especially those occurring outside the known endemic area.
  Parasites & Vectors 05/2013; 6(1):128.
• Article: Expression and cellular trafficking of GP82 and GP90 glycoproteins during Trypanosoma cruzi metacyclogenesis.

  Ethel Bayer-Santos, Narcisa Leal Cunha-E-Silva, Nobuko Yoshida, José Franco da Silveira
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  ABSTRACT: BACKGROUND: The transformation of noninfective epimastigotes into infective metacyclic trypomastigotes (metacyclogenesis) is a fundamental step in the life cycle of Trypanosoma cruzi, comprising several morphological and biochemical changes. GP82 and GP90 are glycoproteins expressed at the surface of metacyclic trypomastigote, with opposite roles in mammalian cell invasion. GP82 is an adhesin that promotes cell invasion, while GP90 acts as a negative regulator of parasite internalization. Our understanding of the synthesis and intracellular trafficking of GP82 and GP90 during metacyclogenesis is still limited. Therefore, we decided to determine whether GP82 and GP90 are expressed only in fully differentiated metacyclic forms or they start to be expressed in intermediate forms undergoing differentiation. METHODS: Parasite populations enriched in intermediate forms undergoing differentiation were analyzed by quantitative real-time PCR, Western blot, flow cytometry and immunofluorescence to assess GP82 and GP90 expression. RESULTS: We found that GP82 and GP90 mRNAs and proteins are expressed in intermediate forms and reach higher levels in fully differentiated metacyclic forms. Surprisingly, GP82 and GP90 presented distinct cellular localizations in intermediate forms compared to metacyclic trypomastigotes. In intermediate forms, GP82 is localized in organelles at the posterior region and colocalizes with cruzipain, while GP90 is localized at the flagellar pocket region. CONCLUSIONS: This study discloses new aspects of protein expression and trafficking during T. cruzi differentiation by showing that the machinery involved in GP82 and GP90 gene expression starts to operate early in the differentiation process and that different secretion pathways are responsible for delivering these glycoproteins toward the cell surface.
  Parasites & Vectors 05/2013; 6(1):127.
• Article: Screening and identification of novel B cell epitopes of Toxoplasma gondii SAG1.

  Yanhua Wang, Guangxiang Wang, Delin Zhang, Hong Yin, Meng Wang
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  ABSTRACT: BACKGROUND: The identification of protein epitopes is useful for diagnostic purposes and for the development of peptide vaccines. In this study, the epitopes of Toxoplasma gondii SAG1 were identified using synthetic peptide techniques with the aid of bioinformatics. FINDINGS: Eleven peptides derived from T. gondii SAG1 were assessed by ELISA using pig sera from different time points after infection. Four (PS4, PS6, PS10 and PS11), out of the eleven peptides tested were recognized by all sera. Then, shorter peptides that were derived from PS4, PS6, PS10 and PS11 were predicted using bioinformatics and tested by experimentation. Four out of nine shorter peptides were identified successfully (amino acids 106--120, 166--180, 289--300 and 313--332). CONCLUSIONS: We have precisely located the epitopes of T. gondii SAG1 using pig sera collected at different time points after infection. The identified epitopes may be useful for the further study of epitope-based vaccines and diagnostic reagents.
  Parasites & Vectors 04/2013; 6(1):125.
• Article: Molecular analysis of Baylisascaris columnaris revealed mitochondrial and nuclear polymorphisms.

  Frits Franssen, Kayin Xie, Hein Sprong, Joke van der Giessen
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  ABSTRACT: BACKGROUND: Baylisascaris species are intestinal nematodes of skunks, raccoons, badgers, and bears belonging to the genus Ascarididae. Oral uptake of embryonated Baylisascaris sp. eggs by a wide variety of mammals and birds can lead to visceral, ocular and neurological larva migrans. B. procyonis, the raccoon roundworm, is known to cause severe illness in intermediate hosts and in humans, whereas the skunk roundworm B. columnaris is probably less pathogenic. Skunks and raccoons are kept as pets in Europe, sometimes together with cats and dogs, living in close contact with humans. B. procyonis and B. columnaris are difficult to differentiate based on morphological criteria and molecular and phylogenetic information concerning B. columnaris is missing. This is the first study on the genetic characterisation of B. columnaris, based on mitochondrial and nuclear molecular markers. METHODS: B. columnaris worms were isolated from pet skunks, and used for molecular analysis. PCR primers targeted at mitochondrial cytochrome c oxidase 1 and 2 (CO1 and CO2), ribosomal ITS1-5.8S-ITS2 and ribosomal 28S genes were used. DNA sequences from B. columnaris, B. procyonis and B. transfuga from bears were analysed by cluster analysis. RESULTS: Four different multi-locus genotypes were found in B. columnaris, based on 14 single nucleotide polymorphisms (SNPs) and two insertions / deletions in CO1, CO2, ITS1-5.8S-ITS2 and 28S. CONCLUSIONS: The genetic characteristics of B. columnaris show close resemblance to those of B. procyonis, but in contrast to B. procyonis, show several polymorphisms in both mitochondrial and nuclear markers. These polymorphisms could be used as a tool to differentiate B. columnaris from B. procyonis in molecular diagnostic assays, and to identify B. columnaris by PCR, in addition to or replacing morphometric analysis. This might lead to more insight into the zoonotic relevance of B. columnaris in humans.
  Parasites & Vectors 04/2013; 6(1):124.
• Article: Paediatric Visceral Leishmaniasis in Italy: a 'One Health' approach is needed.

  Vincenzo Lorusso, Filipe Dantas-Torres, Francesco Caprio, Mariano Manzionna, Francesco Santoro, Gad Baneth, Domenico Otranto
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  ABSTRACT: Here we describe a case of paediatric visceral leishmaniasis recorded in an infant initially suspected for acute lymphoblastic leukaemia due to the clinical and haematological presentation. Eventually the patient was found positive for Leishmania infantum infection and successfully treated. This case emphasises how pivotal a 'One Health' approach is for diagnosing this zoonotic disease; highlighting the importance of including Visceral Leishmaniasis in the differential diagnosis of leukaemia-like syndromes in infants travelling to, and living in, the Mediterranean region.
  Parasites & Vectors 04/2013; 6(1):123.