Five species within the genera Ebolavirus and Marburgvirus of the family Filoviridae are known to cause severe hemorrhagic fever with high mortality rates in humans and non-human primates. Recent large outbreaks of Ebola virus disease in West Africa (2014 -2016) and the Democratic Republic of the Congo (2018 -ongoing) have demonstrated the epidemic potential with devastating public health consequences. Several known and novel filovirus species have been found in bats in recent years. However, the role of each virus species in the disease ecology of human disease is still unclear. In particular, the transmission mechanism from potential animal hosts to humans is not known. Therefore, a simple, flexible, cost-effective screening tool for detecting the presence of any (putative) member of the filovirus family in animal samples is needed. In this study, a one-step conventional pan-filovirus RT-PCR assay was developed. The designed universal consensus primers of this screening test target two highly conserved regions of the nucleoprotein (NP) of all currently known filoviruses. The assay was capable of specific amplification of viral RNA of all six primatepathogenic (human and non-human) filovirus species and resulted in 317 bp long RT-PCR products. This amplicon length renders the assay suitable for flexible application as conventional reverse transcription polymerase chain reaction (RT-PCR) as well as for future use as rapid real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR).

Under controlled test conditions, unfed male and female Hyalomma truncatum ticks exhibited a positive scototaxis to stationary, two-dimensional targets. Upright-positioned rectangles were the most attractive targets. The attractiveness of these targets increased with their size. Significantly more ticks responded scototactically positively to the targets under a luminance contrast ratio of 5:1, as compared with other luminance contrast ratios. Targets with an elevation angle of 13 degrees were occupied more frequently than objects with higher elevation angles. Scototaxis was the same towards a stationary and a sinusoid oscillating target. When an upright-positioned rectangle was combined with a CO2 gradient, the number of ticks that migrated into the CO2 gradient and contacted the target did not increase significantly. The interval between exposure and first locomotion of the ticks, however, was significantly shorter under the influence of a CO2 gradient than in all other experiments without a CO2 gradient. A temperature gradient simulating a natural host (cattle) did not alter the scototaxis. The results of these investigations suggest that the positive scototaxis exhibited by adult H. truncatum ticks is not likely to be part of their appetence behaviour but rather searching behaviour to find adequate protection from harsh climatic conditions.

___________________________________________________________________________First evaluations on field samples, including carnivore feces, animal and human hydatid cyst material from Uganda and Kenya, showed specific amplification of two target regions of the mitochondrial genome of Echinococcus species according to melt and high-resolution melt curve analyses of the developed real-time PCR assays. Consecutive sequencing of PCR products revealed that, apart from Echinococcus felidis, sequences of two other tapeworm species, Echinococcus granulosus sensu stricto and Echinococcus canadensis, which are also endemic in East Africa, were detected by the developed real-time PCR assays. ___________________________________________________________________________ IntroductionThe aim of the present study was to develop a rapid, sensitive, and safe method for molecular detection of Echinococcus felidis, and possibly, related tapeworm species. We first investigated nucleic acid extraction methods suitable for isolating templates directly from fecal samples of carnivores and hydatid cyst material of intermediate hosts in the field without tedious laboratory based enrichment methods, such as digestion or flotation methods. Then, we developed real-time PCR assays which target two different mitochondrial genome regions of E. felidis.