Ethanol inactivation of orthonairoviruses in ixodid ticks.

Ixodid ticks represent vectors and reservoirs for a broad range of zoonotic pathogens. Collected ticks from field studies are therefore usually stored in ethanol, which in higher concentrations effectively inactivates most of the known tick-borne pathogens. Although commonly practiced as gold standard for inactivation, hardly any scientific data demonstrate that ethanol sufficiently penetrates the comparatively thick cuticula of ticks. Therefore, Amblyomma hebraeum tick pools were stored for 21 days in ethanol (96%). Afterwards, the ethanol was removed and the ticks were homogenized. Quantitative 1H-NMR spectroscopic analysis was applied to determine the residual concentration of ethanol inside the ticks. 1H-NMR spectroscopic analysis revealed that ethanol constituted 28.3-42.6 mg of the total weight of three ticks in the pools (89.9-121.5 mg). In addition, the low-pathogenic Hazara orthonairovirus (HAZV) was used as a cell culture model for this study. The virus was exposed to ethanol concentrations between 0 and 60% and incubated under various temperature conditions for four time periods. Afterwards, the residual virus infectivity was determined by titration. Following ethanol exposure, HAZV did not grow in cells after 9 h of exposure to an ethanol concentration of 25%. These results demonstrate an extremely low ethanol resistance of the virus, which was generally in line with previously reported ethanol inactivation data for Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV). After prolonged storage and impregnation, comparable ethanol concentrations are achieved in the ticks, indicating the suitability of this inactivation method also for Bunyaviruses in ticks. At the very least, a massive virus inactivation can be assumed. Definitive proof of virus inactivation would require a bioassay of ethanol-treated infected ticks under appropriate biosafety conditions.

Detection of Crimean-Congo hemorrhagic fever virus in blood-fed Hyalomma ticks collected from Mauritanian livestock.

BACKGROUND: Crimean-Congo hemorrhagic fever virus (CCHFV) belongs to the genus Orthonairovirus (Nairovididae) and is a (re)emerging tick-borne pathogen. It is endemic in most parts of Africa, Asia and southern Europe, and can cause severe hemorrhagic symptoms in humans, with high fatality rates (5-30%). METHODS: Hyalomma ticks were collected from four different livestock herds (cattle and camels) in Mauritania in 2018. The tick species were determined morphologically and confirmed molecularly by using the cytochrome oxidase 1 gene marker. For the detection of CCHFV, ticks were tested individually by one-step multiplex real-time reverse-transcriptase quantitative polymerase chain reaction. The small segment of all positive samples was sequenced to determine the CCHFV genotype. RESULTS: In total, 39 of the 1523 ticks (2.56%) collected from 63 cattles and 28 camels tested positive for CCHFV. Three Hyalomma species were identified. Hyalomma rufipes had the largest proportion of positivity (5.67%; 16/282), followed by Hyalomma dromedarii (1.89%; 23/1214). No Hyalomma impeltatum tested positive (0%; 0/21). Positive ticks were found in only six out of 91 host animals. Viral sequence analysis revealed the presence of two different CCHFV lineages (Africa I and Africa III). CONCLUSIONS: In this study, 2.56% of Hyalomma ticks collected from camels and cattle in Mauritania tested positive for CCHFV. However, the true prevalence of CCHFV in unfed ticks may be lower, as a considerable number of ticks may have been passively infected during blood-feeding by co-feeding ticks or due to viremia of the host. The results indicate the need to track the actual area of circulation of this virus.

Molecular discrimination of Hyalomma tick species serving as reservoirs and vectors for Crimean-Congo hemorrhagic fever virus in sub-Saharan Africa.

The species identification of tick vectors of Crimean-Congo hemorrhagic fever virus (CCHFV), especially Hyalomma (H.) species, is a prerequisite to understand the eco-epidemiology of this disease and to reveal vector and virus reservoir species. However, the morphologic species discrimination can be difficult for damaged or blood-fed ticks and in case of species intercrosses. Therefore, we used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and restriction fragment length polymorphism (RFLP) analysis to distinguish the most common Hyalomma species from sub-Saharan Africa (H. truncatum, H. rufipes and H. dromedarii). Within the last years, MALDI-TOF MS analysis based on tick leg proteins has been shown to be a reliable method to distinguish several tick species. For this purpose, a reference spectral library of several European, American and African tick species was established. In this study, six different Hyalomma species were tested, all of which were all clearly distinguishable by mass spectrometric analyses. Moreover, MALDI TOF- MS was able to confirm morphologic findings where sequencing provided ambiguous results. In addition, a polymerase chain reaction (PCR) based on the CO1 gene amplification of ticks has been developed for the unequivocal species identification by amplicon sequencing and specific restriction endonuclease cleavage pattern analysis. RFLP proved to be a feasible auxiliary discrimination tool for selected Hyalomma species when access to sequencing methods is not available, as for instance during field studies.

Seroprevalence and Associated Risk Factors of Rift Valley Fever in Domestic Small Ruminants in the North Region of Cameroon.

Rift Valley fever (RVF) is a zoonotic vector borne infectious disease of major medical and veterinary importance particularly in sub-Saharan Africa. However, there is dearth of epidemiological knowledge of the disease in Cameroon. We conducted a cross-sectional study (January 2016-January 2017) to investigate the seroprevalence and potential risk factors of Rift Valley fever virus (RVFV) in sheep and goats in the North region of Cameroon. Stratified sampling approach was used to select herds where sera were collected from 680 randomly selected small ruminants (355 goats and 325 sheep) in eight localities (Kismatari, Lagdo, Pitoa, Garoua, Bocklé, Dembo, Poli and Touboro) within three administrative divisions (Bénoué, Mayo-Rey and Faro) in the North region. Anti-RVFV antibodies were detected using a competitive Enzyme-Linked Immunosorbent Assay (ELISA), while a capture ELISA was used for the detection of specific RVFV-Immunoglobulin M (Ig-M) antibodies. We evaluated the associated potential risk factors of RVF in small ruminants based on observations of animal-related intrinsic and extrinsic factors in combination with serological results. The results revealed that 3.4% (95% confidence interval (CI): 2.2-5.1%) of sampled animals and 24.6% (95% CI: 15.1-37.1%) of 65 sampled herds were seropositive for anti-RVFV antibodies and no difference in seropositivity between sheep and goats at individual animal as well as at herd levels was observed. Localities along hydrographic or large water banks such as Kismatari (OR: 14.333, (95% CI: 1.436-145.088)) and Pitoa (OR = 11.467 (95% CI: 1.249-50.306)) were significantly associated to RVFV antibody seroprevalence in a simple logistic regression. In addition, the multiple regression analysis showed that age and access to water points significantly influenced RVFV antibody seroprevalence in small ruminants. This study revealed that anti-RVFV antibodies are present in sheep and goats in the North region of Cameroon. It highlights the likely endemic circulation of RVFV in the considered localities despite the absence of clinical cases reported in animals or humans. Under these conditions, it is necessary to set up an early warning, surveillance and control strategy based on epizootic risk.

Immune recognition of Onchocerca volvulus proteins in the human host and animal models of onchocerciasis.

Onchocerca volvulus is a tissue-dwelling, vector-borne nematode parasite of humans and is the causative agent of onchocerciasis or river blindness. Natural infections of BALB/c mice with Litomosoides sigmodontis and of cattle with Onchocerca ochengi were used as models to study the immune responses to O. volvulus-derived recombinant proteins (OvALT-2, OvNLT-1, Ov103 and Ov7). The humoral immune response of O. volvulus-infected humans against OvALT-2, OvNLT-1 and Ov7 revealed pronounced immunoglobulin G (IgG) titres which were, however, significantly lower than against the lysate of O. volvulus adult female worms. Sera derived from patients displaying the hyperreactive form of onchocerciasis showed a uniform trend of higher IgG reactivity both to the single proteins and the O. volvulus lysate. Sera derived from L. sigmodontis-infected mice and from calves exposed to O. ochengi transmission in a hyperendemic area also contained IgM and IgG1 specific for O. volvulus-derived recombinant proteins. These results strongly suggest that L. sigmodontis-specific and O. ochengi-specific immunoglobulins elicited during natural infection of mice and cattle cross-reacted with O. volvulus-derived recombinant antigens. Monitoring O. ochengi-infected calves over a 26-month period, provided a comprehensive kinetic of the humoral response to infection that was strictly correlated with parasite load and occurrence of microfilariae.

[Evaluation of atrial septum defects in adulthood using echocardiography methods]. / Beurteilung von Vorhofseptumdefekten im Erwachsenenalter mittels echokardiographischer Verfahren.

Two-dimensional and Doppler echocardiographic methods are used to noninvasively detect atrial septal defects. The value of these methods to predict the magnitude of intracardiac left-to-right shunts has not been thoroughly investigated. In this study, we derived right ventricular (RV) and septal defect dimensions, and Qp/Qs-ratios from two-dimensional and Doppler echocardiography in 30 consecutive patients (17 females, 3 males, age 37 +/- 17 years) with invasively confirmed atrial septal defects. Noninvasively obtained parameters were compared to atrial shunt size as measured by oxymetry. RV dimensions correlated only poorly (RV length: r = 0.53, p < 0.005; RV-diameter r = 0.45 p < 0.05), but septal defect dimensions (r = 0.67, p < 0.001) and Qp/Qs-index (r = 0.65, p < 0.05) correlated fairly with shunt size. RV dilatation was highly sensitive (100%) but only moderately specific (67%) as an indicator of shunts > 30%. A defect length > or = 15 mm was moderately sensitive (81%) but highly specific (100%) and a Qp/Qs-index > or = 1.45 was highly sensitive (100%) and specific (76%) to detect shunts > 30%. None of the noninvasive parameters investigated in this study was able to differentiate moderate (> 30% but < 50%) from large ( > or = 50%) intracardiac left-to-right shunts.