Screening of bat faeces for arthropod-borne apicomplexan protozoa: Babesia canis and Besnoitia besnoiti-like sequences from Chiroptera

Background : 45 Microbats (Chiroptera: Microchiropte ra) are among the most eco - epidemiologically important 46 mammals, owing to their presence in human settlements and ani mal keeping facilities . 47 Roosting of bats in buildings may bring pathogens of veterinary - medical importance into the 48 environment of domestic animals and humans. In this context bats have long been studied as 49 carriers of various pathogen groups. However, despite their close association with arthropods 50 (both in their f oo d and as their ectoparasites), only a few molecular surveys have been 51 publish ed on their role as carriers of vector - borne protozoa. The aim of the present study was 52 to compensate for this scarcity of information. 53 Findings : 54 Altogether 221 (mostly individual) bat faecal samples were collected in Hungary and the 55 Netherlands. The DNA w as extracted , and analysed with PCR and sequencing for the 56 presence of arthropod - borne apicomplexan protozoa. Babesia canis canis (with 99 - 100% 57 homology) was identified in five samples, all from Hungary. Because it was excluded with an 58 Ixodidae - specific PC R that the relevant bats consumed ticks, these sequences derive either 59 from insect carriers of Ba. canis , or from the infection of bats. In one bat faecal sample from 60 the Netherlands a sequence having the highest (99%) homology to Besnoitia besnoiti was 61 am plified. 62 Conclusions : 63 The se findings suggest that some aspects of the epidemiology of canine babesiosis are 64 underestimated or unknown, i.e. the potential role of insect - borne mechanical transmission 65 and/or the susceptibility of bats to Ba. canis . In addit ion, b ats need to be added to future 66 studies in the quest for the final host of Be. besnoiti

Latrocimicinae completes the phylogeny of Cimicidae: meeting old morphologic data rather than modern host phylogeny.

The family Cimicidae includes obligate hematophagous ectoparasites (bed bugs and their relatives) with high veterinary/medical importance. The evolutionary relationships of Cimicidae and their hosts have recently been reported in a phylogenetic context, but in the relevant study, one of the six subfamilies, the bat-specific Latrocimicinae, was not represented. In this study the only known species of Latrocimicinae, i.e., Latrocimex spectans, was analyzed with molecular and phylogenetic methods based on four (two nuclear and two mitochondrial) genetic markers. The completed subfamily-level phylogeny of Cimicidae showed that Latrocimicinae is most closely related to Haematosiphoninae (ectoparasites of birds and humans), with which it shares systematically important morphologic characters, but not hosts. Moreover, in the phylogenetic analyses, cimicid bugs that are known to infest phylogenetically distant bat hosts clustered together (e.g., Leptocimex and Stricticimex within Cacodminae), while cimicid subfamilies (Latrocimicinae, Primicimicinae) that are known to infest bat hosts from closely related superfamilies clustered distantly. In conclusion, adding Latrocimicinae significantly contributed to the resolution of the phylogeny of Cimicidae. The close phylogenetic relationship between Latrocimicinae and Haematosiphoninae is consistent with long-known morphologic data. At the same time, phylogenetic relationships of genera within subfamilies are inconsistent with the phylogeny of relevant hosts

Interspecies virus transfer via protoplast fusions between Fusarium poae and black Aspergillus strains

Similarities between the genome organisation of dsRNA mycoviruses and dsRNA patterns in different fungal species suggest a relatedness between these viruses, which could be the result of co-evolved infections or of interspecies transfer. Such interspecies transfer between species is suggested by our observation of transfer and maintenance of mycoviral dsRNAs between Fusarium and Aspergillus via protoplast fusion

Study on the Course of Cryptosporidium baileyi infection in chickens treated with interleukin-1 or indomethacin

The effects exerted by human recombinant interleukin-1β (hrIL-1β) and the prostaglandin inhibitor indomethacin on the course of Cryptosporidium baileyi infection in chickens were studied. Daily oocyst shedding was monitored by a quantitative method throughout the experiment. Humoral immune response to C. baileyi was assessed by ELISA at 3 weeks of age while the level of cellular immune response to phytohaemagglutinin-P (PHA-P) by a skin test at 23 days of age. Parenteral application of hrIL-1b decreased oocyst shedding to 62%, but the infection ran a similar course in treated and control birds. The PHA-P skin test demonstrated increased cellular immune reaction in chickens receiving IL-1b, but there was no significant difference in the humoral responses of the two groups as detected by ELISA. On the other hand, indomethacin mixed to the feed lessened oocyst shedding to 13.7% and also shortened its duration. Immunological parameters as reflected by PHA-P skin test and ELISA results indicated enhanced cellular but unaltered humoral immune response. These data suggest that the sys- temic application of interleukin-1 can induce partial protection against C. baileyi in chickens and that prolonged, abundant oocyst shedding is due to an indometha- cin-sensitive immunodepression via the prostaglandin pathway

Quantification of the relationship between the environment and Fusarium head blight, Fusarium pathogen density, and mycotoxins in winter wheat in Europe

Measurements of local environmental conditions, intensity of Fusarium head blight (FHB) in wheat spikes, biomass of Fusarium graminearum, F. culmorum, and F. poae (pathogens causing FHB) and concentration of the mycotoxins deoxynivalenol (DON) and nivalenol (NIV) in harvested wheat grain were obtained in a total of 150 location-years, originating in three European countries (Hungary, Ireland, United Kingdom) from 2001 to 2004. Through window-pane methodology, the length and starting time of temporal windows where the environmental variables were significantly associated with the biological variables were identified. Window lengths of 5 to 30 days were evaluated, with starting times from 18 days before anthesis to harvest. Associations were quantified with nonparametric Spearman correlation coefficients. All biological variables were significantly associated with at least one evaluated environmental variable (P≤0.05). Moisture-related variables (e.g., average relative humidity, hours of relative humidity above 80%) had the highest positive correlations with the biological variables, but there also was a significant negative correlation between average temperature and several biological variables. When significant correlations were found, they were generally for all window lengths, but for a limited number of window start times (generally before anthesis for disease index and after anthesis for the toxins and late-season fungal biomasses). Semi-partial Spearman correlation coefficients were used to evaluate the relationship between the environmental variables and the concentration of DON and NIV after the effects of FHB intensity and fungal biomass on the mycotoxins were removed. Significant semi-partial correlations were found between relative humidity variables and DON, and between temperature and relative humidity variables and NIV for time windows that started after anthesis (and not for any earlier time windows). Results confirm that the environment influences disease, fungal biomass, and mycotoxin production, and help refine the time windows where the association is greatest. However, variability in the relationships was high, indicating that no single environmental variable is sufficient for prediction of disease or mycotoxin contamination

Q fever epidemic in Hungary, April to July 2013

We investigated a Q fever outbreak with human patients showing high fever, respiratory tract symptoms, headache and retrosternal pain in southern Hungary in the spring and summer of 2013. Seventy human cases were confirmed by analysing their serum and blood samples with micro-immunofluorescence test and real-time PCR. The source of infection was a merino sheep flock of 450 ewes, in which 44.6% (25/56) seropositivity was detected by enzyme-linked immunosorbent assay. Coxiella burnetii DNA was detected by real-time PCR in the milk of four of 20 individuals and in two thirds (41/65) of the manure samples. The multispacer sequence typing examination of C. burnetii DNA revealed sequence type 18 in one human sample and two manure samples from the sheep flock. The multilocus variable-number tandem repeat analysis pattern of the sheep and human strains were also almost identical, 4/5-9-3-3-0-5 (Ms23-Ms24-Ms27-Ms28-Ms33-Ms34). It is hypothesised that dried manure and maternal fluid contaminated with C. burnetii was dispersed by the wind from the sheep farm towards the local inhabitants. The manure was eliminated in June and the farm was disinfected in July. The outbreak ended at the end of July 2013

Diverse tick-borne microorganisms identified in free-living ungulates in Slovakia

Background: Free-living ungulates are hosts of ixodid ticks and reservoirs of tick-borne microorganisms in central Europe and many regions around the world. Tissue samples and engorged ticks were obtained from roe deer, red deer, fallow deer, mouflon, and wild boar hunted in deciduous forests of south-western Slovakia. DNA isolated from these samples was screened for the presence of tick-borne microorganisms by PCR-based methods. Results: Ticks were found to infest all examined ungulate species. The principal infesting tick was Ixodes ricinus, identified on 90.4% of wildlife, and included all developmental stages. Larvae and nymphs of Haemaphysalis concinna were feeding on 9.6% of wildlife. Two specimens of Dermacentor reticulatus were also identified. Ungulates were positive for A. phagocytophilum and Theileria spp. Anaplasma phagocytophilum was found to infect 96.1% of cervids, 88.9% of mouflon, and 28.2% of wild boar, whereas Theileria spp. was detected only in cervids (94.6%). Importantly, a high rate of cervids (89%) showed mixed infections with both these microorganisms. In addition to A. phagocytophilum and Theileria spp., Rickettsia helvetica, R. monacensis, unidentified Rickettsia sp., Coxiella burnetii, "Candidatus Neoehrlichia mikurensis", Borrelia burgdorferi (s.l.) and Babesia venatorum were identified in engorged I. ricinus. Furthermore, A. phagocytophilum, Babesia spp. and Theileria spp. were detected in engorged H. concinna. Analysis of 16S rRNA and groEL gene sequences revealed the presence of five and two A. phagocytophilum variants, respectively, among which sequences identified in wild boar showed identity to the sequence of the causative agent of human granulocytic anaplasmosis (HGA). Phylogenetic analysis of Theileria 18S rRNA gene sequences amplified from cervids and engorged I. ricinus ticks segregated jointly with sequences of T. capreoli isolates into a moderately supported monophyletic clade. Conclusions: The findings indicate that free-living ungulates are reservoirs for A. phagocytophilum and Theileria spp. and engorged ixodid ticks attached to ungulates are good sentinels for the presence of agents of public and veterinary concern. Further analyses of the A. phagocytophilum genetic variants and Theileria species and their associations with vector ticks and free-living ungulates are required.Fil: Kazimírová, Mária. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Hamšíková, Zuzana. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Spitalská, Eva. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; EslovaquiaFil: Minichová, Lenka. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; EslovaquiaFil: Mahríková, Lenka. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Caban, Radoslav. Široká ; EslovaquiaFil: Sprong, Hein. National Institute for Public Health and Environment.Laboratory for Zoonoses and Environmental Microbiology; Países BajosFil: Fonville, Manoj. National Institute for Public Health and Environment.Laboratory for Zoonoses and Environmental Microbiology; Países BajosFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Kocianová, Elena. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; Eslovaqui