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Wild rodents are considered as potential carriers of several zoonotic vector-borne bacteria but their epidemiology is poorly understood in Tunisia. A total of 305 biological samples (100 spleens, 100 livers, 100 kidneys, and 5 pooled ectoparasites (Xenopsylla cheopis, Laelaps echidninus, Ornithonyssus sp., Hoplopleura sp. and eggs of the rat fleas)) were collected from 100 wild rodents from three Tunisian governorates. Molecular screening was performed to reveal infections with main vector-borne bacteria. Captured rodents belonged to three rodent genera and species including Rattus rattus (n = 51, 51%), Meriones shawi (n = 24, 24%) and Mus musculus (n = 25, 25%). Examined rodents were found to be heavily infested by the rat flea X. cheopis (n = 32, 47%) and the rat mite L. echidninus (n = 22, 32.3%). However, the rat mite Ornithonyssus sp. (n = 13, 19.1%) and the rat lice Hoplopleura sp. (n = 1, 1.5%) were rarely identified. Based on 16S rRNA and msp4 genes, infection with Anaplasmataceae bacteria was detected in six specimens of R. rattus and one M. shawi. Pathogenic A. phagocytophilum (n = 1), A. phagocytophilum-like 1 (Anaplasma sp. Japan) (n = 1), and A. ovis (n = 5) were identified. On the basis of ompB, ompA and gltA genes, infection with Rickettsia spp. was identified in three specimens of R. rattus and one of M. shawi. Five Rickettsia species of the spotted fever group, corresponding to R. monacensis, R. helvetica, R. massiliae, R. africae, and R. aeschlimannii, were detected in mixed infections. Bartonella henselae DNA was also found in two R. rattus, based on rpoB partial sequences. All revealed Anaplasma, Rickettsia and Bartonella bacteria were detected in spleen samples. Ehrlichia, Coxiella and Borrelia spp. were not identified in any of the tested samples. In Tunisia, this is the first report indicating infections with Anaplasma, Rickettsia and Bartonella spp. in wild rodents, particularly present alongside domestic livestock and human. This represents a serious risk of potential bacterial transmission. Thus, controlling rodent population in animal herds, residential areas and sensitizing local people to this risk seem absolutely necessary., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The moth pest fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is now present throughout much of the Eastern Hemisphere where it poses a significant economic threat to a number of crops. Native to the Western Hemisphere, fall armyworm is one of the primary pests of corn in the Americas and periodically causes significant economic damage to sorghum, millet, cotton, rice, and forage grasses. This broad host range is in part the result of two populations historically designated as host strains (C-strain and R-strain) that differ in their host plant preferences. Reports of infestations in Africa have to date mostly been limited to the C-strain preferred crops of corn and sorghum, with little evidence of an R-strain presence. However, this could reflect a bias in monitoring intensity, with the R-strain perhaps being more prevalent in other crop systems that have not been as routinely examined for the pest. Because knowledge of whether and to what extent both strains are present is critical to assessments of crops at immediate risk, we analyzed specimens obtained from a systematic survey of pasture grass and rice fields, habitats typically preferred by the R-strain, done contemporaneously with collections from corn fields in Ghana and Togo. Substantial larval infestations were only observed in corn, while pheromone trap capture numbers were high only in corn and rice habitats. Little to no fall armyworm were found in the pasture setting. Comparisons with a meta-analysis of studies from South America identified differences in the pattern of strain-specific markers typically found in fall armyworm collected from rice habitats between the two hemispheres. Genetic tests of specimens from rice and corn area traps failed to show evidence of differential mating between strains. These results are consistent with the R-strain being rare or even absent in Africa and, at least for the Ghana-Togo area, this R-strain lack does not appear to be due to limitations in pest monitoring. The implications of these results to the crops at risk in Africa and the accuracy of existing molecular markers of strain identity are discussed., Competing Interests: The authors have declared that no competing interests exist.

Through international trades, Europe, Africa and South America share a long history of exchanges, potentially of pathogens. We used the worldwide parasite Toxoplasma gondii to test the hypothesis of a historical influence on pathogen genetic diversity in Benin, a West African country with a longstanding sea trade history. In Africa, T. gondii spatial structure is still non-uniformly studied and very few articles have reported strain genetic diversity in fauna and clinical forms of human toxoplasmosis so far, even in African diaspora. Sera from 758 domestic animals (mainly poultry) in two coastal areas (Cotonou and Ouidah) and two inland areas (Parakou and Natitingou) were tested for T. gondii antibodies using a Modified Agglutination Test (MAT). The hearts and brains of 69 seropositive animals were collected for parasite isolation in a mouse bioassay. Forty-five strains were obtained and 39 genotypes could be described via 15-microsatellite genotyping, with a predominance of the autochthonous African lineage Africa 1 (36/39). The remaining genotypes were Africa 4 variant TUB2 (1/39) and two identical isolates (clone) of Type III (2/39). No difference in terms of genotype distribution between inland and coastal sampling sites was found. In particular, contrarily to what has been described in Senegal, no type II (mostly present in Europe) was isolated in poultry from coastal cities. This result seems to refute a possible role of European maritime trade in Benin despite it was one of the most important hubs during the slave trade period. However, the presence of the Africa 1 genotype in Brazil, predominant in Benin, and genetic analyses suggest that the triangular trade was a route for the intercontinental dissemination of genetic strains from Africa to South America. This supports the possibility of contamination in humans and animals with potentially imported virulent strains., Competing Interests: The authors have declared that no competing interests exist.

The worldwide presence of Leishmania parasites increases in the poorest regions. Current leishmaniasis treatments are unsatisfactory due to resistance development, side effects and cost. Herein, we describe the in vitro activity of artemisinin (ART), artemether (ATM), artesunate (ATS) and dihydroartemisinin (DHA) against Leishmania amazonensis. Selected compounds were assayed in the animal model of cutaneous leishmaniasis in BALB/c mice. On intracellular amastigotes, similar activity (p > 0.05) was observed for ART, ATM and ATS (IC 50  = 15.0-19.2 μM), which were inferior (p < 0.05) respect to reference endoperoxide ascaridole (IC 50  = 11.5 ± 1.0 μM) and superior (p < 0.05) compared with reference drug Glucantime® (IC 50  = 30.1 ± 9.0 μM). In contrast, DHA (IC 50  = 38.5 ± 4.7 μM) showed higher IC 50 values (p < 0.05) than other artemisinins and ascaridole, but similar (p > 0.05) than Glucantime®; while deoxyartemisinin caused smaller inhibition (IC 50  = 88.9 ± 5.2 μM). Selectivity indexes of >13, 6, 11 and 1 were obtained for ART, ATM, ATS and DHA, respectively. In addition, the potential effect of ART and ATS was also demonstrated in the murine model, causing a significant reduction (p < 0.05) of the lesion size and parasite load regarding untreated animals and treated with vehicle. Effects of both artemisinins were comparable (p > 0.05) with Glucantime® and ascaridole-treated mice. In particular, artemisinin is recommended to further studies, which could be an advantage over the ascaridole endoperoxide and could be useful in endemic areas of parasite resistance to antimonials., (Copyright © 2020. Published by Elsevier B.V.)

Introduction: Hammondia hammondi and Toxoplasma gondii are closely related protozoan parasites, but only T. gondii is zoonotic. Both species use felids as definitive hosts and cannot be differentiated by oocyst morphology. In T. gondii, a 529-base pair (bp) repetitive element (TgREP-529) is of utmost diagnostic importance for polymerase chain reaction (PCR) diagnostic tests. We identified a similar repetitive region in the H. hammondi genome (HhamREP-529)., Methods: Based on reported sequences, primers and probes were selected in silico and optimal primer probe combinations were explored, also by including previously published primers. The analytical sensitivity was tested using serial dilutions of oocyst DNA. For testing analytical specificity, DNA isolated from several related species was used as controls. The newly established TaqMan PCR (Hham-qPCR1) was applied to tissues collected from H. hammondi-infected gamma-interferon gene knockout (GKO) mice at varying time points post-infection., Results: Ten forward and six reverse primers were tested in varying combinations. Four potentially suitable dual-labelled probes were selected. One set based on the primer pair (Hham275F, Hham81R) and the probe (Hham222P) yielded optimal results. In addition to excellent analytic specificity, the assay revealed an analytical sensitivity of genome equivalents of less than one oocyst. Investigation of the tissue distribution in GKO mice revealed the presence of parasite DNA in all examined organs, but to a varying extent, suggesting 100- to 10,000-fold differences in parasitic loads between tissues in the chronic state of infection, 42 days post-infection., Discussion: The use of the 529-bp repeat of H. hammondi is suitable for establishing a quantitative real-time PCR assay, because this repeat probably exists about 200 times in the genome of a single organism, like its counterpart in T. gondii. Although there were enough sequence data available, only a few of the primers predicted in silico revealed sufficient amplification; the identification of a suitable probe was also difficult. This is in accord with our previous observations on considerable variability in the 529-bp repetitive element of H. hammondi., Conclusions: The H. hammondi real-time PCR represents an important novel diagnostic tool for epidemiological and cell biological studies on H. hammondi and related parasites.

Fighting trypanosomiasis with an anti-trypanosome vaccine is ineffective, the parasite being protected by a Variable Surface Glycoprotein (VSG) whose structure is modified at each peak of parasitaemia, which allows it to escape the host's immune defenses. However, the host immunization against an essential factor for the survival of the parasite or the expression of its pathogenicity could achieve the same objective. Here we present the results of mouse immunization against the Translationally Controlled Tumor Protein (TCTP), a protein present in the Trypanosoma brucei gambiense (Tbg) secretome, the parasite responsible for human trypanosomiasis. Mice immunization was followed by infection with Tbg parasites. The production of IgG, IgG1 and IgG2a begun after the second TCTP injection and was dose-dependant, the maximum level of anti-TCTP antibodies remained stable up to 4 days post-infection and then decreased. Regarding cytokines (IL-2, 4, 6, 10, INFγ, TNFα), the most striking result was their total suppression after immunization with the highest TCTP dose. Compared to the control group, the immunized mice displayed a reduced first peak of parasitaemia, a 100% increase in the time to onset of the second peak, and an increased time of mice survival. The effect of immunization was only transient but demonstrated the likely important role that TCTP plays in host-parasite interactions and that some key parasite proteins could reduce infection impact., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Klossiella is a genus of apicomplexan coccidian parasites with a global distribution, whose members typically infect the renal tissue of a wide variety of vertebrate hosts with a high level of host specificity. The presence of this parasite has been previously associated with kidney inflammatory processes. To our knowledge, this is the first report on the prevalence of Klossiella muris in the house mouse (Mus musculus) in Portugal (São Miguel Island - Azores). The prevalence of K. muris was determined through histopathological examination of renal tissue collected during necropsy of 130 mice captured between the years of 2011-2019. K. muris was diagnosed in 45.38% (CI 95 : 40.9-85.4) of the examined mice. Infection with this parasite was associated with mild to severe kidney inflammation, assessed by the presence of inflammatory processes in the renal cortex and medulla. CAPSULE: First record on coccidiosis caused by infection of Klossiella muris in Mus musculus in Portugal., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Because isolated ecosystems contribute to species variability, especially oceanic island ecosystems, the present work focused on the study of the Bartonella species and haplotypes in Lanzarote and El Hierro, two Canary islands with evident bioclimatic differences between them. A total of 123 rodents and 110 fleas from two islands were screened for the presence of Bartonella by PCR analysis of the gltA and nuoG genes. The overall prevalence was 5.7% in rodents and 20.4% in fleas. A total of seven gltA-haplotypes was found in both rodents and fleas, belonging to the species Bartonella mastomydis and Bartonella tribocorum in Lanzarote, and to Bartonella rochalimae and Bartonella elizabethae in El Hierro, as well as recently described species Bartonella kosoyi in both islands. Besides, potential co-infections were detected based on the nuoG analysis. Further, Xenopsylla cheopis was the only flea species identified. Our study shows that isolated ecosystems such as the Canary Islands lead to the appearance of new Bartonella haplotypes along different biotopes, with diverse flea species involved in the spreading of the pathogen being of great relevance due to the zoonotic potential of the species found., (© 2020 The Society for Vector Ecology.)

Objectives: Progress in national schistosomiasis control in China has successfully reduced disease transmission in many districts. However, a low transmission rate hinders conventional snail surveys in identifying areas at risk. In this study, Schistosoma japonicum-infected sentinel mice surveillance was conducted to identify high-risk areas of schistosomiasis transmission in Hubei province, China., Methods: The risk of schistosomiasis transmission was assessed using sentinel mice monitoring in Hubei province from 2010 to 2018. Field detections were undertaken in June and September, and the sentinel mice were kept for approximately 35 days in a laboratory. They were then dissected to determine whether schistosome infection was present. Ripley's K-function and kernel density estimation were applied to analyze the spatial distribution and positive point pattern of schistosomiasis transmission., Results: In total, 190 sentinel mice surveillance sites were selected to detect areas of schistosomiasis infection from 2010 to 2018, with 29 (15.26%) sites showing infected mice. Of 4723 dissected mice, 256 adult worms were detected in 112 infected mice. The infection rate was 2.37%, with an average of 2.28 worms detected per infected mouse. Significantly more infected mice were detected in the June samples than in the September samples (χ 2 =12.11, p<0.01). Ripley's L(d) index analysis showed that, when the distance was ≤34.52km, the sentinel mice infection pattern showed aggregation, with the strongest aggregation occurring at 7.86km. Three hotspots were detected using kernel density estimation: at the junction of Jingzhou District, Gong'an County, and Shashi District in Jingzhou City; in Wuhan City at the border of the Huangpi and Dongxihu Districts, and in the Hannan and Caidian Districts., Conclusion: The results showed that sentinel mice surveillance is useful in identifying high-risk areas, and could provide valuable information for schistosomiasis prevention and control, especially concerning areas along the Yangtze River, such as the Fu-Lun, Dongjing-Tongshun, and Juzhang River basins., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Detection methods for Demodex musculi were historically unreliable, and testing was rarely performed because its prevalence in laboratory mice was underestimated. Although infestations are unapparent in most mouse strains, D. musculi burdens are higher and clinical signs detected in various immunodeficient strains. The parasite's influence on the immune system of immunocompetent mice is unknown. We characterized mite burden (immunocompetent and immunodeficient strains) and immunologic changes (immunocompetent strains only) in naïve Swiss Webster (SW; outbred), C57BL/6NCrl (B6; Th1 responder), BALB/cAnNCrl (BALB/c; Th2 responder) and NOD. Cg- Prkdc scid Il2rg tm1Wjl /SzJ (NSG; immunodeficient) mice after exposure to Demodex- infested NSG mice. Infested and uninfested age-matched mice of each strain ( n = 5) were euthanized 14, 28, 56, and 112 d after exposure. Mite burden was determined through PCR analysis and skin histopathology; B-cell and CD4 + and CD8 + T-cell counts and activation states (CD25 and CD69) were evaluated by using flow cytometry; CBC counts were performed; and serum IgE levels were measured by ELISA. Mite burden and PCR copy number correlated in NSG mice, which had the highest mite burden, but not in immunocompetent strains. Infested immunocompetent animals developed diffuse alopecia by day 112, and both BALB/c and C57BL/6 mice had significantly increased IgE levels. These findings aligned with the skewed Th1 or Th2 immunophenotype of each strain. BALB/c mice mounted the most effective host response, resulting in the lowest mite burden of all immunocompetent strains at 112 d after infestation without treatment. Clinically significant hematologic abnormalities were absent and immunophenotype was unaltered in immunocompetent animals. Topical treat- ment with imidacloprid-moxidectin (weekly for 8 wk) was effective at eradicating mites by early as 7 d after treatment. IgE levels decreased substantially in infested BALB/c mice after treatment. These findings demonstrate a need for D. musculi surveillance in mouse colonies, because the infestation may influence the use of infested mice in select studies.

Rodents are reservoirs and hosts of several pathogens around the world, including zoonotic parasite species. This study aimed to determine the occurrence of zoonotic gastrointestinal helminths in rodents captured inside households in a rural community from southern Guatemala. Sixty-nine rodents were captured in 33% (49/148) of the surveyed households, including Rattus rattus, Rattus norvegicus, Mus musculus, and Sigmodon hispidus. Thirty-six percent (25/69) of these rodents (3 Rattus and 22 Mus musculus), from 45% (22/49) of the households, were parasitized with at least 1 gastrointestinal helminth species. Helminths from 6 species were identified: Hymenolepis diminuta, Moniliformis moniliformis, Heterakis spumosa, Nippostrongylus sp., Strongyloides sp., and Syphacia sp. Two zoonotic species were found in Rattus, H. diminuta in R. rattus (1/6), and M. moniliformis in R. norvegicus (1/1). Coinfection with other non-zoonotic helminth parasites, such as He. spumosa and Strongyloides sp., also was observed in the Rattus genus. Mus musculus had only non-zoonotic helminths: He. spumosa, Nippostrongylus sp., and Syphacia sp. being the most common, and He. spumosa (96%) followed by Nippostrongylus sp. (48%), with a higher presence in males than females, with a similar proportion in adult and young individuals. This is the first report of zoonotic and non-zoonotic helminths parasites in rodents from Guatemala., (© American Society of Parasitologists 2020.)

Genetic diversity in animal immune systems is usually beneficial. In hybrid recombinants, this is less clear, as the immune system could also be impacted by genetic conflicts. In the European house mouse hybrid zone, the long-standing impression that hybrid mice are more highly parasitized and less fit than parentals persists despite the findings of recent studies. Working across a novel transect, we assessed infections by intracellular protozoans, Eimeria spp., and infections by extracellular macroparasites, pinworms. For Eimeria, we found lower intensities in hybrid hosts than in parental mice but no evidence of lowered probability of infection or increased mortality in the centre of the hybrid zone. This means ecological factors are very unlikely to be responsible for the reduced load of infected hybrids. Focusing on parasite intensity (load in infected hosts), we also corroborated reduced pinworm loads reported for hybrid mice in previous studies. We conclude that intensity of diverse parasites, including the previously unstudied Eimeria, is reduced in hybrid mice compared to parental subspecies. We suggest caution in extrapolating this to differences in hybrid host fitness in the absence of, for example, evidence for a link between parasitemia and health., (© 2019 The Authors. Journal of Evolutionary Biology published by John Wiley & Sons Ltd on behalf of European Society for Evolutionary Biology.)

Toxoplasma gondii is a ubiquitous highly prevalent zoonotic protozoan. Cats are the definitive hosts, while all other warm-blooded animals are intermediate hosts for this parasite. Commensal rodents, being the main prey of cats, are probably the major reservoir for T. gondii in the domestic environment. Rodents can acquire infection after ingestion of oocysts that have sporulated in the environment. However, experimental evidence shows that vertical transmission can be sufficient for the perpetuation of transmission between generations of mice. In natural settings, the relative epidemiological importance of vertical transmission over oral transmission is a matter of debate and raises the question of the possibility of a T. gondii cycle in the absence of cats. In the present study, we took advantage of an extensive survey of commensal rodents in Dakar, Senegal, where the house mouse is the predominant putative reservoir of T. gondii. Mice genotypes and spatial location through GPS referencing of all trapping localizations were investigated in relation to T. gondii infection in eight sites of the city of Dakar and on Goree Island. In each sampling site, the occurrence of over-prevalence zones of T. gondii infection was investigated through Kulldorf's statistic using SaTScan software. Genetic structure and relatedness between mice were investigated within each over-prevalence zone, in order to find clues of transmission between related mice. Within each of the four over-prevalence zones identified across nine sites, infected mice belonged to more than one genetic group. No association between the degree of relatedness and the occurrence of T. gondii infection could be detected. These findings suggest an environmental source of infection for mice associated with localized putative foci of environmental contamination and support an oral route of infection for mice from Dakar rather than a cycle based on vertical transmission. However, further investigations based on a denser sampling in different epidemiological contexts are recommended., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest related to this publication., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The role of rodents as reservoirs of helminths of public health importance is not well known. The zoonotic potential of Syphacia spp. has been confirmed; therefore, the study aimed to estimate the occurrence of oxyurid nematodes in small rodents from pet shops and breeding clubs in Slovakia. Fecal samples of 586 pet rodents kept in 133 cages were collected between 2016 and 2018 and examined by Faust´s flotation method. Four species of oxyurid nematodes, Syphacia muris, S. obvelata, Aspiculuris tetraptera and Paraspidodera uncinata were detected. A. tetraptera was found in the faecal samples of all rodent species included in this survey. The number of positive boxes varied from 5.4% in hamsters to 70.0% with mice. The prevalence of Syphacia muris was highest in Mongolian gerbils where up to 75.0% boxes were positive; S. obvelata was found in 26.7% of boxes with mice, 25.0% of boxes with Mongolian gerbils and 3.2% of boxes with rats. The high prevalence of Syphacia spp. in all animal species points out the infection risk for humans. Animals offered for sale are often in close contact with human beings; therefore they should be regularly tested for parasites and then effectively dewormed.

Introduction: Trypanosoma cruzi is the causal agent of the American trypanosomiasis, an endemic disease in México. The commensal rodents Mus musculus and Rattus rattus are reservoirs of this parasite, which invades cardiac fibers and develops parasite nests causing various lesions. Histopathological studies in naturally infected rodents are scarce. Objective: To describe the types and frequencies of microscopic lesions in cardiac tissue of M. musculus and R. rattus infected with T. cruzi captured in Mérida, México. Materials and methods: The rodents were captured in suburban environments of Mérida. Cardiac tissue was extracted and processed by the paraffin inclusion technique and hematoxylin and eosin stained. The observation was made with a conventional microscope and all the lesions, as well as their degree, were identified. Results: Eight tissue samples of M. musculus and seven of R. rattus were studied. Parasite nests were found in 7/15, specifically 3/8 in M. musculus and 4/7 in R. rattus. The inflammatory infiltrate was the most frequent lesion. Other lesions were: Degeneration of cardiac fibers (8/15), congestion of blood vessels (6/15), and necrosis (5/15). Discussion: The lesions we observed have been described in experimental animal models and in humans with American trypanosomiasis. The inflammatory infiltrate has been identified as the most significant lesion in humans and reservoirs in the chronic stage of the disease. Conclusion: The lesions we described are associated with T. cruzi infection, which confirms that the rodents studied are reservoirs of this parasite.

Toxoplasma gondii is a zoonotic protozoan with a worldwide occurrence, but the determinants of the current pattern in the geographical distribution of T. gondii lineages and strains remain poorly understood. To test the influence of human trade on T. gondii populations, we conducted a population genetic study of 72 T. gondii animal isolates from Senegal, a West African country in which the ongoing inland progress of invasive murine hosts (introduced in port cities of Senegal since the 16th century by European sailors) is well described. Isolates were mainly collected on free-range poultry, which are considered as relevant bioindicators of T. gondii strain diversity in the domestic environment. Sampling was conducted in two port cities of Senegal (Dakar and Saint-Louis) and in one inland region (Kedougou). Population genetic analyses using 15 microsatellite markers revealed different patterns between port cities where lineages non-virulent for mice (type II, type III, and Africa 4) were predominant, and Kedougou where the mouse-virulent Africa 1 lineage was the most common. By considering the current spatial pattern in the inland progress of invasive rodents in Senegal, our results suggest that the invasive house mouse Mus musculus domesticus counter-selects the Africa 1 lineage in the invaded areas. The comparison of the microsatellite alleles of type II strains from Senegal to type II strains from other areas in Africa and Western Europe, using discriminant analysis of principal components and Network analysis, point to a mainly Western European origin of the type II lineage in Senegal. Collectively, these findings suggest that human-mediated intercontinental migrations of murine hosts are important vectors of T. gondii strains. Differential susceptibility of endemic and introduced murine hosts to various T. gondii strains probably determines the persistence of these strains in the environment, and therefore their availability for human and animal infection., Competing Interests: The authors have declared that no competing interests exist.

An inverse relationship between biodiversity and human health has been termed the 'dilution effect' paradigm. In the case of tick-borne infections such as Lyme disease, the key assumption is that Borrelia burgdorferi sensu lato abundance is increased by the loss of less competent (dilution) hosts as biodiversity declines. White-tailed deer play a dual role in the pathogen cycle, as key reproductive hosts for adult ticks and incompetent hosts for the pathogen. While the role of deer as hosts of adult ticks is well established, the extent to which deer also feed immature ticks and reduce the proportion infected is unknown because of logistic constraints in measuring this empirically. We estimated the proportion of larvae that fed on deer in an extremely species-poor community on Block Island, RI, where tick nymphal infection prevalence was found to be lower than expected. In 2014, we measured the density, larval tick burdens, and realized reservoir competence of small mammal and bird hosts on Block Island, RI. In 2015, we measured the infection prevalence of host-seeking Ixodes scapularis nymphs resulting from larvae fed on available hosts in 2014. We back-estimated the proportion of larvae expected to have fed on deer in 2014 (the only unknown parameter) to result in the nymphal infection prevalence observed in 2015. Back-estimation predicted that 29% of larval ticks must have fed on deer to yield the observed 30% nymphal infection prevalence. In comparison, the proportion of larvae feeding on mice was 44% and 27% on birds. Our study identified an influential role of deer in reducing nymphal tick infection prevalence and a potential role as dilution hosts if the reduction in nymphal infection prevalence outweighs the role of deer as tick population amplifiers. Because both deer and competent hosts may increase in anthropogenic, fragmented habitats, the links between fragmentation, biodiversity, and Lyme disease risk may be complex and difficult to predict. Furthermore, a nonlinear relationship between deer abundance and Lyme disease risk would reduce the efficacy of deer population reduction efforts to control Lyme disease., (Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.)

The severe virulence of Toxoplasma gondii in classical laboratory inbred mouse strains contradicts the hypothesis that house mice (Mus musculus) are the most important intermediate hosts for its transmission and evolution because death of the mouse before parasite transmission equals death of the parasite. However, the classical laboratory inbred mouse strains (Mus musculus domesticus), commonly used to test Toxoplasma strain differences in virulence, do not capture the genetic diversity within Mus musculus. Thus, it is possible that Toxoplasma strains that are severely virulent in laboratory inbred mice are avirulent in some other mouse sub-species. Here, we present insight into the responses of individual mouse strains, representing strains of the genetically divergent Mus musculus musculus, Mus musculus castaneus and Mus musculus domesticus, to infection with individual clonal and atypical Toxoplasma strains. We observed that, unlike M. m. domesticus, M. m. musculus and M. m. castaneus are resistant to the clonal Toxoplasma strains. For M. m. musculus, we show that this is due to a locus on chromosome 11 that includes the genes that encode the interferon gamma (IFNG)-inducible immunity-related GTPases (Irgs) that can kill the parasite by localising and subsequently vesiculating the parasitophorous vacuole membrane. However, despite the localization of known effector Irgs to the Toxoplasma parasitophorous vacuole membrane, we observed that some atypical Toxoplasma strains are virulent in all the mouse strains tested. The virulence of these atypical strains in M. m. musculus could not be attributed to individual rhoptry protein 5 (ROP5) alleles, a secreted parasite pseudokinase that antagonises the canonical effector Irgs and is indispensable for parasite virulence in laboratory inbred mice (M. m. domesticus). We conclude that murine resistance to Toxoplasma is modulated by complex interactions between host and parasite genotypes and may be independent of known effector Irgs on murine chromosome 11., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Diagnosis of parasitic diseases that involve tissue-stage larvae is challenging, and serology remains the most effective antemortem test for detecting these infections. Baylisascaris procyonis, the raccoon roundworm, is a zoonotic ascarid. Raccoons are the usual definitive host, and humans may be infected as accidental hosts. More than 150 species of birds and mammals may act as paratenic hosts, and rodents play an important role in the transmission and maintenance of this parasite in nature. Migratory larvae in paratenic host tissues can produce ocular disease and severe to fatal neurologic disease, but not all infected hosts develop signs. A sensitive and specific Western blot (WB) assay based on a recombinant Baylisascaris-specific antigen (rBpRAG-1) has been developed for use in humans. We evaluated the use of this antigen to detect Baylisascaris spp. infections in rodent paratenic hosts. With the use of 4 species of Peromyscus mice ( Peromyscus californicus, Peromyscus leucopus, Peromyscus maniculatus, Peromyscus polionotus) from a previous infection trial, we developed species-adapted WB and ELISA assays and evaluated performance compared to detection of larvae in tissue samples. These assays revealed species-level differences in seroconversion and terminal antibody concentrations, with P. leucopus developing significantly greater antibody concentrations than P. californicus and P. polionotus at all dose levels, and P. maniculatus at the low dose. Some P. californicus and P. polionotus failed to seroconvert despite the recovery of larvae from their tissues. WB and ELISA results were correlated; however, the WB demonstrated higher sensitivity than the ELISA overall (72.2% versus 63.9%, respectively). With the use of experimental samples, specificity was 100% for WB and 94.1% for ELISA. A WB was also used to test Mus and Rattus samples, and although numbers were too limited to evaluate sensitivity and specificity, all animals known to be infected by tissue digestion were WB positive, and all uninfected animals were negative. Finally, the Peromyscus-adapted WB and ELISA were used to test a set of serum samples from wild-trapped P. maniculatus and Rattus rattus. Both assays were generally sensitive, but specificity was equivocal. This emphasizes the challenge of using serology for investigation of wildlife diseases, in which hosts have unknown exposure histories. Nevertheless, serologic methods have utility in the study of Baylisascaris spp. in paratenic hosts, either wild or captive, and have advantageous attributes (non-lethal, high-throughput), but results should be interpreted carefully.

Parasite hybrid zones resulting from host secondary contact have never been described in nature although parasite hybridization is well known and secondary contact should affect them similarly to free-living organisms. When host populations are isolated, diverge and recontact, intimate parasites (host specific, direct life cycle) carried during isolation will also meet and so may form parasite hybrid zones. If so, we hypothesize these should be narrower than the host's hybrid zone as shorter parasite generation time allows potentially higher divergence. We investigate multilocus genetics of two parasites across the European house mouse hybrid zone. We find each host taxon harbours its own parasite taxa. These also hybridize: Parasite hybrid zones are significantly narrower than the host's. Here, we show a host hybrid zone is a suture zone for a subset of its parasite community and highlight the potential of such systems as windows on the evolutionary processes of host-parasite interactions and recombinant pathogen emergence., (© 2018 John Wiley & Sons Ltd.)

Background: Nicotinamide adenine dinucleotide (NAD+) is an essential molecule in the energy metabolism of living beings, and it has various cellular functions. The main enzyme in the biosynthesis of this nucleotide is nicotinamide/nicotinate mononucleotide adenylyltransferase (NMNAT, EC 2.7.7.1/18) because it is the convergence point for all known biosynthetic pathways. NMNATs have divergences in both the number of isoforms detected and their distribution, depending on the organism., Methods: In the laboratory of basic research in biochemistry (LIBBIQ: acronym in Spanish) the NMNATs of protozoan parasites (Leishmania braziliensis, Plasmodium falciparum, Trypanosoma cruzi, and Giardia duodenalis) have been studied, analysing their catalytic properties through the use of proteins. Recombinants and their cellular distribution essentially. In 2014, O'Hara et al. determined the cytoplasmic localization of NMNAT of P. falciparum, using a transgene coupled to GFP, however, the addition of labels to the study protein can modify several of its characteristics, including its sub-cellular localization., Results: This study confirms the cytoplasmic localization of this protein in the parasite through recognition of the endogenous protein in the different stages of the asexual life cycle. Additionally, the study found that PfNMNAT could be a phosphorylation target at serine, tyrosine and threonine residues, and it shows variations during the asexual life cycle., Conclusions: These experiments confirmed that the parasite is situated in the cytoplasm, fulfilling the required functions of NAD+ in this compartment, the PfNMNAT is regulated in post-transcription processes, and can be regulated by phosphorylation in its residues.

Background: Increasing resistance to current anti-malarial therapies requires a renewed effort in searching for alternative therapies to combat this challenge, and combination therapy is the preferred approach to address this. The present study confirms the anti-plasmodial effects of two compounds, cryptolepine and xylopic acid and the relationship that exists in their combined administration determined., Methods: Anti-plasmodial effect of cryptolepine (CYP) (3, 10, 30 mg kg -1 ) and xylopic acid (XA) (3, 10, 30 mg kg -1 ) was evaluated in Plasmodium berghei-infected male mice after a 6-day drug treatment. The respective doses which produced 50% chemosuppression (ED 50 ) was determined by iterative fitting of the log-dose responses of both drugs. CYP and XA were then co-administered in a fixed dose combination of their ED 50 s (1:1) as well as different fractions of these combinations (1/2, 1/4, 1/8, 1/16 and 1/32) to find the experimental ED 50 (Z exp ). The nature of interaction between cryptolepine and xylopic acid was determined by constructing an isobologram to compare the Z exp with the theoretical ED 50 (Z add ). Additionally, the effect of cryptolepine/xylopic acid co-administration on vital organs associated with malarial parasiticidal action was assessed., Results: The Z add and Z exp were determined to be 12.75 ± 0.33 and 2.60 ± 0.41, respectively, with an interaction index of 0.2041. The Z exp was significantly (P < 0.001) below the additive isobole indicating that co-administration of cryptolepine and xylopic acid yielded a synergistic anti-plasmodial effect. This observed synergistic antiplasmodial effect did not have any significant deleterious effect on the kidney, liver and spleen. However, the testis were affected at high doses., Conclusion: The co-administration of cryptolepine and xylopic acid produces synergistic anti-malarial effect with minimal toxicity.

Background: Malaria is an infectious disease caused by parasites of the genus Plasmodium, of which Plasmodium vivax and Plasmodium falciparum are the major species that cause the disease in humans. As there are relatively few alternatives for malaria treatment, it is necessary to search for new chemotherapeutic options. Colombia possesses a great diversity of plants, which are potential sources of new compounds of medical interest. Thus, in this study the antiplasmodial effect of extracts from two species of plants from the families Simaroubaceae and Picramniaceae (Picramnia latifolia and Picrolemma huberi) was evaluated in vitro and in vivo. These plants were chosen because they contain secondary metabolites with interesting medicinal effects., Results: The ethanolic extracts of both species were highly active with IC 50 : 1.2 ± 0.19 µg/mL for P. latifolia and IC 50 : 0.05 ± 0.005 µg/mL for P. huberi. The P. latifolia extract had a stage specific effect on trophozoites and inhibited parasite growth in vivo by 52.1 ± 3.4%, evaluated at 1000 mg/kg in Balb/c mice infected with Plasmodium berghei. On the other hand, evaluated at 150 mg/kg body weight in the same murine model, the ethanolic extract from P. huberi had an antiplasmodial effect in all the asexual intraerythrocytic stages of P. falciparum FCR3 and inhibited the parasitic growth in 93 ± 32.9%., Conclusions: This is the first report of anti-malarial activity for these two species of plants. Thus, P. latifolia and P. huberi are potential candidates for the development of new drugs for treating malaria.

Background: In the context of malaria elimination/eradication, drugs that are effective against the different developmental stages of the parasite are highly desirable. The oldest synthetic anti-malarial drug, the thiazine dye methylene blue (MB), is known for its activity against Plasmodium blood stages, including gametocytes. The aim of the present study was to investigate a possible effect of MB against malaria parasite liver stages., Methods: MB activity was investigated using both in vitro and in vivo models. In vitro assays consisted of testing MB activity on Plasmodium falciparum, Plasmodium cynomolgi and Plasmodium yoelii parasites in human, simian or murine primary hepatocytes, respectively. MB in vivo activity was evaluated using intravital imaging in BALB/c mice infected with a transgenic bioluminescent P. yoelii parasite line. The transmission-blocking activity of MB was also addressed using mosquitoes fed on MB-treated mice., Results: MB shows no activity on Plasmodium liver stages, including hypnozoites, in vitro in primary hepatocytes. In BALB/c mice, MB has moderate effect on P. yoelii hepatic development but is highly effective against blood stage growth. MB is active against gametocytes and abrogates parasite transmission from mice to mosquitoes., Conclusion: While confirming activity of MB against both sexual and asexual blood stages, the results indicate that MB has only little activity on the development of the hepatic stages of malaria parasites.

Background: Trypanosoma cruzi is the agent of Chagas disease, a major public health problem in Latin America. Many wild and domestic animals are naturally infected with T. cruzi; rodents are one of the groups which have been consistently detected infected in different countries. The aim of this work was to characterize blood T. cruzi load in naturally infected rodents from a Chagas disease endemic region in Chile., Methods: Baited traps were set in domestic and peridomestic areas of rural dwellings. The rodents were anesthetized and blood sampled; DNA was extracted and the parasite load was quantified by T. cruzi satellite DNA real-time PCR assays., Results: Seventy-one rodents of four species, Rattus rattus, Mus musculus, Phyllotis darwini and Octodon degus, were captured; R. rattus was the most abundant species. Fifty-nine samples (83.1%) were T. cruzi-positive and the median value of the parasite load was 2.99 parasite equivalents (par-eq)/ml. The comparison of frequency of infection or parasite load by species showed no differences. However, one R. rattus presented very elevated parasitemia (1644 par-eq/ml)., Conclusions: The overall levels of parasitemia were similar to those found in humans in Chile. The high infection levels in exotic and endemic rodents very near to rural settlements increases their relevance as T. cruzi hosts.

Genetic and environmental factors shape host susceptibility to infection, but how and how rapidly environmental variation might alter the susceptibility of mammalian genotypes remains unknown. Here, we investigate the impacts of seminatural environments upon the nematode susceptibility profiles of inbred C57BL/6 mice. We hypothesized that natural exposure to microbes might directly (e.g., via trophic interactions) or indirectly (e.g., via microbe-induced immune responses) alter the hatching, growth, and survival of nematodes in mice housed outdoors. We found that while C57BL/6 mice are resistant to high doses of nematode (Trichuris muris) eggs under clean laboratory conditions, exposure to outdoor environments significantly increased their susceptibility to infection, as evidenced by increased worm burdens and worm biomass. Indeed, mice kept outdoors harbored as many worms as signal transducer and activator of transcription 6 (STAT6) knockout mice, which are genetically deficient in the type 2 immune response essential for clearing nematodes. Using 16S ribosomal RNA sequencing of fecal samples, we discovered enhanced microbial diversity and specific bacterial taxa predictive of nematode burden in outdoor mice. We also observed decreased type 2 and increased type 1 immune responses in lamina propria and mesenteric lymph node (MLN) cells from infected mice residing outdoors. Importantly, in our experimental design, different groups of mice received nematode eggs either before or after moving outdoors. This contrasting timing of rewilding revealed that enhanced hatching of worms was not sufficient to explain the increased worm burdens; instead, microbial enhancement and type 1 immune facilitation of worm growth and survival, as hypothesized, were also necessary to explain our results. These findings demonstrate that environment can rapidly and significantly shape gut microbial communities and mucosal responses to nematode infections, leading to variation in parasite expulsion rates among genetically similar hosts.

Syphacia stroma (von Linstow, 1884) Morgan, 1932 and Syphacia frederici Roman, 1945 are oxyurid nematodes that parasitize two murid rodents, Apodemus sylvaticus and Apodemus flavicollis, on the European mainland. Only S. stroma has been recorded previously in Apodemus spp. from the British Isles. Despite the paucity of earlier reports, we identified S. frederici in four disparate British sites, two in Nottinghamshire, one each in Berkshire and Anglesey, Wales. Identification was based on their site in the host (caecum and not small intestine), on key morphological criteria that differentiate this species from S. stroma (in particular the tail of female worms) and by sequencing two genetic loci (cytochrome C oxidase 1 gene and a section of ribosomal DNA). Sequences derived from both genetic loci of putative British S. frederici isolates formed a tight clade with sequences from continental worms known to be S. frederici, clearly distinguishing these isolates from S. stroma which formed a tight clade of its own, distinct from clades representative of Syphacia obvelata from Mus and S. muris from Rattus. The data in this paper therefore constitute the first record of S. frederici from British wood mice, and confirm the status of this species as distinct from both S. obvelata and S. stroma.

We undertook a study on Cryptosporidium spp. in wild cricetid rodents. Fecal samples were collected from meadow voles (Microtus pennsylvanicus), southern red-backed voles (Myodes gapperi), woodland voles (Microtus pinetorum), muskrats (Ondatra zibethicus) and Peromyscus spp. mice in North America, and from bank voles (Myodes glareolus) and common voles (Microtus arvalis) in Europe. Isolates were characterized by sequence and phylogenetic analyses of the small subunit ribosomal RNA (SSU) and actin genes. Overall, 33·2% (362/1089) of cricetids tested positive for Cryptosporidium, with a greater prevalence in cricetids from North America (50·7%; 302/596) than Europe (12·1%; 60/493). Principal Coordinate analysis separated SSU sequences into three major groups (G1-G3), each represented by sequences from North American and European cricetids. A maximum likelihood tree of SSU sequences had low bootstrap support and showed G1 to be more heterogeneous than G2 or G3. Actin and concatenated actin-SSU trees, which were better resolved and had higher bootstrap support than the SSU phylogeny, showed that closely related cricetid hosts in Europe and North America are infected with closely related Cryptosporidium genotypes. Cricetids were not major reservoirs of human pathogenic Cryptosporidium spp.

Some small mammals occur as household pests and harbour a number of parasites that could be of public health importance. This study profiled the helminth and protozoan parasites in trapped small mammals within and around human dwelling places (houses) located across 4 major towns (Auchi, Benin, Ekpoma, and Uromi) and environs in Edo state, Nigeria. Six genera ( Apodemus sp., Crocidura sp., Mastomys natalensis , Mus musculus , Rattus sp., and Sorex sp.) were identified from 502 trapped small mammals. Overall, M. musculus (71.9%) and Rattus rattus (20.1%) were the most frequently trapped. In total, on examination of blood, gastrointestinal contents, and brain tissues, 12 helminth taxa ( Angiostrongylus sp., Aspicularis sp., Capillaria sp., Gongylonema sp., Heterakis spumosa, Hymenolepis diminuta, Hymenolepis nana, Mastophorus muris, Moniliformis moniliformis, Nippostrongylus brasiliensis, Strongyloides sp., Trichosomoides sp., and Trichuris sp.), and 6 protozoan parasites ( Babesia sp., Trypanosoma lewisi, Plasmodium sp., Eimeria sp., Isospora sp., and Toxoplasma gondii ) were isolated. Most prevalent helminths with relatively heavy mean intensity were Strongyloides sp. and Heterakis spumosa , while Plasmodium, Eimeria , and Isospora were the most prevalent protozoan parasites. Generally, intrinsic factors like sex and age had marginal influence on the rate and burden of infection in M. musculus and R. rattus . Although the infection rate and prevalence of zoonotic parasites were low, they were largely recovered in rodents from Ekpoma. This study elucidates the public health implication of the presence of zoonotic parasites in these small mammals.

Rodents are the main reservoir animals of leptospirosis. In this study, we characterized and quantified the urinary excretion dynamics of Leptospira by Mus musculus infected with 2 × 10 8 virulent Leptospira borgpetersenii serogroup Ballum. Each micturition was collected separately in metabolic cages, at 12 time points from 7 to 117 days post-infection (dpi). We detected Leptospira in all urine samples collected (up to 8 per time point per mouse) proving that Leptospira excretion is continuous with ca. 90% live L. borgpetersenii Ballum, revealed by viability quantitative polymerase chain reaction. Microscopic visualization by Live/Dead fluorescence confirmed this high proportion of live bacteria and demonstrated that L. borgpetersenii Ballum are excreted, at least partly, as bacterial aggregates. We observed two distinct phases in the excretion dynamics, first an increase in Leptospira concentration shed in the urine between 7 and 63 dpi followed by a plateau phase from 63 dpi onward, with up to 3 × 10 7 Leptospira per mL of urine. These two phases seem to correspond to progressive colonization of renal tubules first, then to stable cell survival and maintenance in kidneys. Therefore, chronically infected adult mice are able to contaminate the environment via urine at each micturition event throughout their lifetime. Because Leptospira excretion reached its maximum 2 months after infection, older rodents have a greater risk of contaminating their surrounding environment.

Objective To investigate the water infectivity in key water regions, and analyze the transmission risk of schistosomiasis in suspicious and high-risk environments in Hubei Province in 2016. Methods Schistosome-endemic areas of the Yangtze River, the Hanbei River and the Fu River were chosen as the surveillance and forecast sites. The water infectivity was detected by using the sentinel mice during the flood season. The infection status of residents in the villages around the surveillance sites and the activities of human beings and domestic animals were surveyed. The emergency response system was initiated when the water infectivity areas were detected. Results From May to June, among the ten surveillance sites of the first batch, two positive spots with infected mice were found, accounting for 20%. All the recovered mice (totally 200) were dissected: five mice were infected, with a total infection rate of 2.5%. Totally five schistosome adult worms were collected, with mean worm burden of 1 worm per infected mouse. The site with cercariae-infected water body started the emergency response and no epidemic occurred. From August to September, among the eight surveillance sites of the second batch, the recovered mice (totally 160) were dissected, and no infected sentinel mice were found. Conclusion The sentinel mice method plays an important role in analyzing the schistosomiasis transmission risk in Hubei Province.

Tick infestations on small mammals were studied from April to November, 2010, in deciduous woodland in southern England in order to determine whether co-infestations with tick stages occurred on small mammals, a key requirement for endemic transmission of tick-borne encephalitis virus (TBEV). A total of 217 small mammals was trapped over 1,760 trap nights. Yellow-necked mice (Apodemus flavicollis) made up the majority (52.5%) of animals, followed by wood mice (A. sylvaticus) 35.5% and bank voles (Myodes glareolus) 12%. A total of 970 ticks was collected from 169 infested animals; 96% of ticks were Ixodes ricinus and 3% I. trianguliceps. Over 98% of ticks were larval stages. Mean infestation intensities of I. ricinus were significantly higher on A. flavicollis (6.53 ± 0.67) than on A. sylvaticus (4.96 ± 0.92) and M. glareolus (3.25 ± 0.53). Infestations with I. ricinus were significantly higher in August than in any other month. Co-infestations with I. ricinus nymphs and larvae were observed on six (3.6%) infested individuals, and fifteen small mammals (8.9%) supported I. ricinus - I. trianguliceps co-infestations. This work contributes further to our understanding of European small mammal hosts that maintain tick populations and their associated pathogens, and indicates that co-infestation of larvae and nymph ticks does occur in lowland UK. The possible implications for transmission of tick-borne encephalitis virus between UK ticks and small mammals are discussed., (© 2017 The Society for Vector Ecology.)

Background: The liver stages of Plasmodium parasites are important targets for the discovery and development of prophylactic drugs., Methods: A real-time in vivo imaging system was used to determine the level of luminescence measured from firefly luciferase expression by sporozoites developing in hepatocytes in different strains of mice., Results: The luminescence values (photon counts/sec) measured from the anatomical liver location in the untreated mice infected with 10,000 Plasmodium berghei sporozoites were 8.15 × 10 5 for C57BL/6 Albino, 2.12 × 10 5 for C3H/HeNCrL, 0.91 × 10 5 for C57BL/6 WT, 0.28 × 10 5 for BALB/c, and 0.16 × 10 5 for ICR/CD-1 mice. This data suggests that the C57BL/6 Albino strain is most susceptible to luminescent photon, mainly because the less light scattering and absorption from deeper tissues and the skin in the strain of mouse. The photon count observed in black C57BL/6 wild type mice was shown to be 88.83% lower compared to C57BL/6 Albino mice. Although the highest growth rate of sporozoites in hepatocytes was found for C57BL/6 wild type mice in this study, the black skin of this mouse significantly reduced parasite-associated bioluminescence., Conclusions: The minimal light scattering and absorption and also enhanced susceptibility to liver infection of C57BL/6 Albino mice makes this strain preferable sensitivity for discovery and development of prophylactic antimalarial drugs., (Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.)

Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10 7 tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Understanding why some exotic species become widespread and abundant in their colonised range is a fundamental issue that still needs to be addressed. Among many hypotheses, newly established host populations may benefit from a parasite loss ("enemy release" hypothesis) through impoverishment of their original parasite communities or reduced infection levels. Moreover, the fitness of competing native hosts may be negatively affected by the acquisition of exotic taxa from invaders ("parasite spillover") and/or by an increased transmission risk of native parasites due to their amplification by invaders ("parasite spillback"). We focused on gastrointestinal helminth communities to determine whether these predictions could explain the ongoing invasion success of the commensal house mouse (Mus musculus domesticus) and black rat (Rattus rattus), as well as the associated decrease in native Mastomys spp., in Senegal. For both invasive species, our results were consistent with the predictions of the enemy release hypothesis. A decrease in overall gastrointestinal helminth prevalence and infracommunity species richness was observed along the invasion gradients as well as lower specific prevalence/abundance (Aspiculuris tetraptera in Mus musculus domesticus, Hymenolepis diminuta in Rattus rattus) on the invasion fronts. Conversely, we did not find strong evidence of GIH spillover or spillback in invasion fronts, where native and invasive rodents co-occurred. Further experimental research is needed to determine whether and how the loss of gastrointestinal helminths and reduced infection levels along invasion routes may result in any advantageous effects on invader fitness and competitive advantage., (Copyright © 2016 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

The cat flea, Ctenocephalides felis felis (Bouché), feeds on different host species, causing annoyance or transmitting disease agents. In this study, the influence of the host of the cat flea on blood feeding, egg production, and sex ratio of the offspring was investigated. Two strains of C. felis were domesticated on either rats or mice for >10 yr in the laboratory, and in this study, these fleas were placed in the following groups and fed on rats or mice continuously: Group A (rat-domesticated C. felis with rats as host); Group B (rat-domesticated C. felis with mice as host); Group C (mouse-domesticated C. felis with rats as host); and Group D (mouse-domesticated C. felis with mice as host). In total, 240 adult fleas were in each group at a sex ratio of female:male = 1.7:1. The mean egg production per flea of Groups A, B, C, and D was 55.0, 19.2, 62.5, and 13.2, respectively. A significant correlation between egg production and the volume of blood consumed was detected for Groups A, B, C, and D. The sex ratio (F:M) of the offspring in Groups A and C was 2.07 and 2.11, respectively, whereas in Groups B and D, the ratio was 1.04 and 1.03, respectively. In conclusion, the C. felis with rats as host consumed more blood, produced more eggs, and had higher sex ratios of the offspring than those with mice as the host., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Demodex musculi, a prostigmatid mite that has been reported infrequently in laboratory mice, has been identified with increasing frequency in contemporary colonies of immunodeficient mice. Here we describe 2 episodes of D. musculi infestation with associated clinical signs in various genetically engineered mouse strains, as well as treatment strategies and an investigation into transmissibility and host susceptibility. The first case involved D. musculi associated with clinical signs and pathologic lesions in BALB/c-Tg(DO11.10)Il13(tm) mice, which have a defect in type 2 helper T cell (Th2) immunity. Subsequent investigation revealed mite transmission to both parental strains (BALB/c-Tg[DO11.10] and BALB/c-Il13(tm)), BALB/c-Il13/Il4(tm), and wild-type BALB/c. All Tg(DO11.10)Il13(tm) mice remained infested throughout the investigation, and D. musculi were recovered from all strains when they were cohoused with BALB/c-Tg(DO11.10)Il13(tm) index mice. However, only Il13(tm) and Il13/Il4(tm) mice demonstrated persistent infestation after index mice were removed. Only BALB/c-Tg(DO11.10)Il13(tm) showed clinical signs, suggesting that the phenotypic dysfunction of Th2 immunity is sufficient for persistent infestation, whereas clinical disease associated with D. musculi appears to be genotype-specific. This pattern was further exemplified in the second case, which involved NOD.Cg-Prkdc(scid)Il2r(tm1Wjl)/SzJ (NSG) and C;129S4 Rag2(tm1.1Flv) Il2rg(tm1.1Flv)/J mice with varying degrees of blepharitis, conjunctivitis, and facial pruritis. Topical amitraz decreased mite burden but did not eliminate infestation or markedly ameliorate clinical signs. Furthermore, mite burden began to increase by 1 mo posttreatment, suggesting that topical amitraz is an ineffective treatment for D. musculi. These experiences illustrate the need for vigilance regarding opportunistic and uncommon pathogens in rodent colonies, especially among mice with immunologic deficits.

Detection of Toxoplasma gondii DNA in cat feces is considered indicative of the presence of T. gondii oocysts. This study aims to demonstrate that the high sensitivity of qPCR can lead to T. gondii DNA detection in cat feces in the absence of oocysts. A cat immune to toxoplasmosis was fed with a mouse experimentally infected with T. gondii. Detection of DNA of this parasite was performed by qPCR on feces passed: (i) on the day the cat ingested the infected prey; (ii) during the three previous days; and (iii) during the three following days. The kinetics of qPCR results are clearly not linked to oocyst shedding and this result demonstrates that qPCR can detect T. gondii DNA related to bradyzoites from an infected prey, in the absence of oocysts. Caution is thus recommended when interpreting T. gondii qPCR results for samples of cat feces., (© M.-L. Poulle et al., published by EDP Sciences, 2016.)

The reed vole Microtus fortis is the only known mammal in which the schistosome is naturally prevented from maturing and schistosome infection does not cause significant pathogenesis. However, the mechanism behind this phenomenon remains unknown. In the present study, Solexa deep sequencing technology was used to carry out high-throughput sequencing and comparative analysis of microRNA (miRNA) between small RNA libraries isolated from 10 days oldschistosomula of M. fortis and BALB/c mice.In total, 10d schistosomula from M. fortis and BALB/c mice yielded 13.37 and 10.84 million reads, respectively, and nearly 39% and 40% of reads could be mapped to selected miRNAs in miRbase. Based on a bioinformatic analysis, we found that most of the miRNAs identified in Schistosoma japonicum were detected in our study. Further analysis revealed that 24 miRNAs were differentially expressed between the schistosomula from the two rodents, of which 21 were down-regulated and three were up-regulated in schistosomula from M. fortis. Also, six novel miRNAs were predicted and identified in this study. Target genes were mapped and filtered by correlating them with differentially expressed genes obtained from S. japonicum oligonucleotide microarray analyses performed in previous studies. miRNAs such as miR-10-3p, miR-10-5p, and miR-2b-5p may affect the growth, differentiation, and metabolism of worms via regulation of the expression of target genes such as enolase, aquaporin, TGF-beta-inducible nuclear protein, and paramyosin. Gene Ontology analysis of the predicted target genes of these six differentially expressed miRNAs revealed that some important biological pathways, such as metabolic processes,glycolysis, and catalytic activity, were involved. The results of this study highlight the function of miRNAs in the development and survival of the schistosome, and provide valuable information to increase our understanding of the regulatory function of miRNAs in schistosome development and host-parasite interactions in a differentially susceptible host environment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

We carried out the first survey of Hymenolepis spp. infection in pet rodents in Italy. Fresh fecal samples were collected from 172 pet rodents as follows: guinea pigs (Cavia porcellus; n = 60), squirrels (Callosciurus finlaysonii, Callosciurus prevosti, Tamias striatus, Tamias sibiricus, Sciurus calorinensis; n = 52), hamsters (Phodopus campbelli, Mesocricetus auratus; n = 30), chinchillas (Chinchilla lanigera; n = 13), rats (Rattus norvegicus; n = 10), and mice (Mus minutoides; n = 7). These animals were housed either in pet shops or in private houses. All fecal samples were processed using the FLOTAC pellet technique to assess the number of eggs per gram (EPG) of feces. Eggs of Hymenolepis nana were found in 24 out of 172 (13.9 %; 95 % confidence interval = 9.3-20.2 %) pet rodents. Of those rodents, 41.6 % (10/24) were rats (mean EPG = 55.7; range = 2-200), 29.2 % (7/24) mice (mean EPG = 64.5; range = 32-120), 25.0 % (6/24) were chinchillas (mean EPG = 25.5; range = 10-50), and 4.2 % (1/24) hamsters (P. campbelli) (EPG = 86.0). In addition, Hymenolepis diminuta eggs were found in 2 out of 172 (1.2 %; 95 % confidence interval = 0.2-4.6 %) rodents examined, both of which (100 %; 2/2) were pet squirrels (C. prevosti) (mean EPG = 10; range = 4-16). To the authors' knowledge, this is the first report of a natural infection of H. diminuta in pet squirrels.

Schistosomiasis is a parasitic disease caused by flatworms of the genus Schistosoma. Among the Schistosoma species known to infect humans, S. mansoni is the most frequent cause of intestinal schistosomiasis in sub-Saharan Africa and South America: the World Health Organization estimates that about 200,000 deaths per year result from schistosomiasis in sub-Saharan Africa alone. The Schistosoma life cycle requires two different hosts: a snail as intermediate host and a mammal as definitive host. People become infected when they come into contact with water contaminated with free-living larvae (e.g. when swimming, fishing, washing). Although S. mansoni has mechanisms for escaping the host immune system, only a minority of infecting larvae develop into adults, suggesting that strain selection occurs at the host level. To test this hypothesis, we compared the Belo Horizonte (BH) strain of S. mansoni recovered from definitive hosts with different immunological backgrounds using random amplification of polymorphic DNA-polymerase chain reaction (RAPD-PCR). Schistosoma mansoni DNA profiles of worms obtained from wild-type (CD1 and C57BL/6J) and mutant (Jα18- / - and TGFβRIIdn) mice were analysed. Four primers produced polymorphic profiles, which can therefore potentially be used as reference biomarkers. All male worms were genetically distinct from females isolated from the same host, with female worms showing more specific fragments than males. Of the four host-derived schistosome populations, female and male adults recovered from TGFβRIIdn mice showed RAPD-PCR profiles that were most similar to each other. Altogether, these data indicate that host immunological backgrounds can influence the genetic diversity of parasite populations.

The gut microbiota profoundly affects the biology of its host. The composition of the microbiota is dynamic and is affected by both host genetic and many environmental effects. The gut microbiota of laboratory mice has been studied extensively, which has uncovered many of the effects that the microbiota can have. This work has also shown that the environments of different research institutions can affect the mouse microbiota. There has been relatively limited study of the microbiota of wild mice, but this has shown that it typically differs from that of laboratory mice (and that maintaining wild caught mice in the laboratory can quite quickly alter the microbiota). There is also inter-individual variation in the microbiota of wild mice, with this principally explained by geographical location. In this study we have characterised the gut (both the caecum and rectum) microbiota of wild caught Mus musculus domesticus at three UK sites and have investigated how the microbiota varies depending on host location and host characteristics. We find that the microbiota of these mice are generally consistent with those described from other wild mice. The rectal and caecal microbiotas of individual mice are generally more similar to each other, than they are to the microbiota of other individuals. We found significant differences in the diversity of the microbiotas among mice from different sample sites. There were significant correlations of microbiota diversity and body weight, a measure of age, body-mass index, serum concentration of leptin, and virus, nematode and mite infection.

This article describes two new skin mite species found on the house mouse Mus musculus L., 1758. Demodex marculus sp. nov. is a very small demodecid mite (adult stages, on average, 99 µm in length) found in mouse skin in the abdomen, back, limbs, and anal area. It is characterized by relatively large bossing hammer-shaped supracoxal spines, embedded in the trapezoidal gnathosoma. Demodex fusiformis sp. nov., in turn, is a little larger (adult stages on average 111 µm in length), with a small oval gnathosoma equipped with fine, knob-like supracoxal spines. It was found in the skin of abdomen, back, and limbs. Moreover, Demodex musculi (Oudemans, 1897) was redescribed, which is small demodecid mite (adult stages on average 142 µm in length) and characterized by relatively large morphological variation and considerable sexual dimorphism. The characteristic feature of this species is the strongly elongated and rectangular gnathosoma equipped with very large wedge-shaped supracoxal spines. D. musculi was found in the skin of various, haired regions of the mice body (head, neck, abdomen, back, limbs, genital-anal region, and tail). Moreover, one more demodecid mite was found in the skin of the examined mice, it was Demodex flagellurus Bukva, 1985, which was found only in the genital area. Overall infection of Mus musculus L. by all species of Demodex was with the prevalence of 100%, mean intensity of 24.0, and range of intensity of 1-109. Despite high infection levels, no symptoms of parasitosis were observed in the hosts., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

A new species Demodex conicus n. sp. is described based on adult and juvenile stages from the ear canals of the house mouse Mus musculus L. in Poland. The new species is most similar to D. auricularis Izdebska, Rolbiecki & Fryderyk, 2014 from the ear canals of the wood mouse Apodemus sylvaticus (L.), but differs in the following features: the gnathosoma is triangular, the supracoxal spines (setae elc.p) are conical, the spines on the terminal segment of palp are four, the striation on opisthosoma is fine but dense, the vulva is located at a distance of c.17 µm from posterior level of legs IV, and the male genital opening is located at the level of legs I. The differences also relate to body size and proportions, female D. conicus n. sp. being, on average slightly larger, and male significantly larger than D. auricularis. Males of the new species also have longer and more massive opisthosoma than males of D. auricularis. Demodex conicus n. sp. was found in 17.5% of the mice studied from different locations in Poland.

Bioinformatics approaches were used to identify B-cell epitopes and T-cell epitopes on SAG5A protein. Compared to SAG1, SAG5A with good B-cell epitopes and T-cell epitopes had a potentiality to become a more successful vaccine against Toxoplasma gondii. Thereafter, SAG5A DNA vaccine was constructed successfully and was injected into mice with peptide to evaluate the immunoprotection. Compared to the control groups, the vaccine (DNA/peptide) could induce more effective cellular and humoral immune responses in immunized mice. Furthermore, a significant reduction of brain cyst was detected in the mice vaccinated with peptide (732±160), pSAG5A (815±197), or pSAG5A/peptide (436±174) compared by the mice injected by PBS (1260±241) or pEGFP-C1 (1350±268). The number of cysts in brains was 35% reduced in the mice immunized with DNA/peptide than in the control mice treated by PBS. The results indicated that the DNA vaccine encoding SAG5A significantly induced immune responses and enhanced protection against cysts of PRU strain, especially with the help of peptide., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Objective: To establish and perfect the surveillance and forecast system of schistosomiasis, and timely discover the suspicious high risk environments for preventing the human and livestock from schistosomiasis., Methods: Eight villages of three counties were selected as survey points. Then, the surveillance and forecast of sentinel mice were carried out in the key water regions. The recovered sentinel mice were dissected in laboratory. The sentinel mouse serum antibodies against schistosome were detected by ELISA, the suspicious water contacts of residents and livestock were investigated and the results were analyzed, and the epidemic risk was assessed., Results: Totally 300 sentinel mice were placed, the recovery rate was 94.67%, and the mortality rate was 8.80%. There were no mice with positive serum antibodies against schistosome, and the results of the dissection of all the sentinel mice were negative. The humans who contacted with the suspicious water were mainly villagers, students, children and fishermen by washing hands and feet, washing vegetables, harvesting, fishing and swimming. The Oncomelania hupensis snail areas, human infection rates, and cattle infection rates were obviously declined in recent 3 years. However, the epidemic risks still existed., Conclusion: Although schistosomiasis transmission was effectively controlled in the three counties, the comprehensive control measures still should be strengthened.

Parasites can generate complex life history trade-offs in a host. In this study, we experimentally reduced the infection level of intestinal helminth parasites in the Taiwan field mouse (Apodemus semotus) to test (1) whether parasite richness and load are biased towards male or female mice (sex-biased parasitism) and (2) whether the effects of parasitism on the host's survival and reproduction are different between the sexes (sex-specific effects of parasitism). Our findings indicate that neither parasite richness (number of helminth taxa found in a fecal sample) nor parasite load (number of helminth eggs per gram of fecal material) was sexually biased in our A. semotus study population. These results are in agreement with those of previous studies on endoparasites in Apodemus spp., but are in contrast to those on ectoparasites in Apodemus spp. Parasite removal reduced the survival rate of reproducing females, possibly by allowing reproducing females to increase maternal investment in their current litters at the cost of their own future survival. Single-litter mothers with reduced parasitism had a higher body mass than the untreated single-litter mothers, suggesting an increased maternal investment. In addition, the reproductively more active A. semotus, particularly the females, carried higher parasite loads, suggesting a trade-off between reproduction and parasite defense. By demonstrating that parasites can affect life history trade-offs in A. semotus, our results highlight the importance of maintaining variation in life history traits under parasitism risks and illustrate the subtle demographic processes (e.g. reduced future survival among healthy reproducing females) that might be driven by parasitism.

We examined changes to the behaviour of flour beetles, Tribolium confusum, infected with the rodent stomach worm, the spirurid Protospirura muricola, in the context of the 'Behavioural Manipulation Hypothesis'. Trobolium confusum infected with the third-stage infective larvae of P. muricola showed consistently altered patterns of behaviour. Relative to uninfected beetles, over a measured time period, beetles infected with P. muricola were likely to move over a shorter distance, when moving their speed of movement was slower, they were more likely to stay in the illuminated area of their environment, more likely to emerge from darkened areas into the illuminated areas, and their longevity was significantly shortened. The changes in behaviour, as reflected in effects on speed of movement, were only evident among beetles that actually harboured infective cysts and not among those carrying younger infections when the larvae within their haemocoels would have been at an earlier stage of development and not yet capable of infecting the definitive murine hosts. We discuss whether these changes would have made the beetles more susceptible to predation by rodents, and specifically by the omnivorous eastern spiny mouse, Acomys dimidiatus, the natural definitive host of this parasite in Egypt, from where the P. muricola isolate originated, and whether they support the Behavioural Manipulation Hypothesis or reflect parasite-induced pathology.

Background: Synanthropic rats and mice share the same environment with humans and play an important role in epidemiology of toxoplasmosis; however, there is limited information about prevalence and genetic characterization of Toxoplasma gondii in synanthropic rats and mice in China., Findings: In the present study, the prevalence and genetic characterization of T. gondii naturally infected synanthropic rodents (Rattus norvegicus and Mus musculus) were investigated in the urban area of Xuzhou city, Eastern China between June 2013 and August 2014. DNA from the brain of each animal was prepared and screened by specific PCR assay targeting 35-fold repeated B1 gene (B1-PCR). PCR positive DNA samples were further genotyped by multi-locus PCR-RFLP. Overall, out of 123 synanthropic rodents, 29 samples were positive by B1 gene-targeted PCR (23.6%). Of these, 7 out of 31 (22.3%) M. musculus were positive, whereas the positive rate of R. norvegicus was 23.9% (22/92). Multi-locus PCR-RFLP analysis reveals that seven PCR-positive samples were completely genotyped and they were identified as type China 1 (ToxoDB# 9)., Conclusion: To our knowledge, this is the first report of molecular detection and genetic characterization of T. gondii infection in synanthropic rodents in Eastern China. The results of the present study showed a high infection pressure of T. gondii exists in the environment and synanthropic rodents infected by T. gondii may be an important source of infection for cats and other animals.

Seven laboratory mammal and bird species were orally inoculated with 200-1,000 encysted Metagonimus hakubaensis metacercariae that had been isolated from naturally infected lampreys (Lethenteron reissneri) captured in Aomori Prefecture. At 8 and 15 days post-infection, adult flukes were recovered from all of the laboratory animals tested, and therefore, hamster, rat, mouse, dog, cat, chicken and quail were considered as final hosts of M. hakubaensis. Recovery rates of the fluke were higher in dogs and hamsters than in cats, rats, mice, chickens and quails. The flukes recovered from dogs and hamsters showed increased body length and higher fecundity than those recovered from the other hosts. These results indicate that the suitability of dogs and hamsters for M. hakubaensis infection is higher than that of the other laboratory animals.