Crimean-Congo Hemorrhagic Fever Virus in Ticks Collected from Cattle, Corsica, France, 2023.

We report the detection of Crimean-Congo hemorrhagic fever virus (CCHFV) in Corsica, France. We identified CCHFV African genotype I in ticks collected from cattle at 2 different sites in southeastern and central-western Corsica, indicating an established CCHFV circulation. Healthcare professionals and at-risk groups should be alerted to CCHFV circulation in Corsica.

Comparison of Crimean-Congo Hemorrhagic Fever Virus and Aigai Virus in Life Cycle Modeling Systems Reveals a Difference in L Protein Activity.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne orthonairovirus that causes a severe, often fatal, hemorrhagic disease throughout Africa, Asia, and Southeast Europe. A wide variety of strains are circulating in the field which broadly correlate to their geographic distribution. The viral determinants of pathogenicity remain unclear, as does the contribution of strain-specific differences to pathology. Aigai virus (AIGV) is a closely related virus (formally designated CCHFV genotype VI, Europe II, or AP92-like virus), which has been proposed to be less virulent than CCHFV. However, the molecular details leading to potential differences in virulence are unknown. To explore if differences exist, life cycle modeling systems, including both a minigenome and a transcriptionally competent virus-like particle assay, were developed for AIGV to allow the comparison with the CCHFV reference IbAr10200 strain. Using this approach, we could demonstrate that AIGV exhibits lower viral gene expression than the reference strain of CCHFV. Subsequent systematic exchange of viral components revealed that the L protein is responsible for the observed differences in gene expression and that the interferon (IFN) antagonistic activity of the ovarian tumor-type protease domain is not responsible for this effect. IMPORTANCE Crimean-Congo hemorrhagic fever virus (CCHFV) is the cause of severe hemorrhagic disease, which is often fatal. Present throughout Africa, Asia, and Southeast Europe, a diverse number of viral genotypes exist. However, the viral determinants of pathogenicity remain unclear. It has been proposed that the closely related Aigai virus (AIGV) may be a less virulent virus. Here, using newly developed and improved life cycle modeling systems we have examined potential differences between the CCHFV reference strain, IbAr10200, and AIGV. Using this approach, we identified lower viral gene expression driven by the AIGV viral polymerase as a major difference which may be indicative of lower virulence.

Sequential determination of viral load, humoral responses and phylogenetic analysis in fatal and non-fatal cases of Crimean-Congo hemorrhagic fever patients from Gujarat, India, 2019.

BACKGROUND: Thirty-four CCHF cases (17 fatal; 17 survived) were confirmed from Gujarat state, India during the year 2019. We aimed to find out the viral load, antibody kinetics, cytokine profile and phylogenetic analysis between fatal and non- fatal cases. METHODS: Thirty four cases were included in this study. Blood and urine samples were collected from all the cases on the day of admission to hospital. Non-fatal cases were followed weekly for understanding the profile of viral kinetics, anti-CCHFV IgM and IgG antibodies. We also quantified the cytokines in both fatal and non-fatal cases. For epidemiological correlation, livestock were screened for anti-CCHF IgG antibodies and the tick pool specimens were tested by real time RT-PCR. Virus isolation was attempted on tick pools and human specimens and phylogenetic analysis performed on human and ticks complete genome sequences. RESULTS: CCHF cases were detected throughout year in 2019 with the peak in August. Out of 34 cases, eight secondary CCHF cases were reported. Cases were predominantly detected in males and in 19-45 years age group (55.88%). The persistence of viremia was observed till 76th POD (post onset date) in one case whereas anti-CCHFV IgM and IgG was detected amongst these cases from the 2nd and 20th POD respectively. Positivity observed amongst livestock and tick pools were was 21.57% and 7.4% respectively. The cytokine analysis revealed a significant increase in the level of serum IL-6, IL-10 and IFN-γ during the acute phase of the infection, but interestingly IL-10 lowered to normal upon clearance of the virus in the clinically recovered case. Fatal cases had high viral RNA copy numbers. Bleeding from one or two mucosal sites was significantly associated with fatality (OR-16.47;p-0.0034 at 95% CI). We could do CCHF virus isolation from two cases. Phylogenetic analysis revealed circulation of re-assortment of Asian-West African genotypes in humans and ticks. CONCLUSIONS: The persistence of CCHF viral RNA was detected till 76th POD in one of the survivors. The circulation of a re-assortment Asian-West African genotype in a CCHF case is also reported first time from India.

Prevalence and phylogenetic analysis of Crimean-Congo hemorrhagic fever virus in ticks collected from Punjab province of Pakistan.

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease of human that caused by CCHF virus. To study the epidemiological distribution of CCHFV, 2183 tick samples were collected from sheep, goats, cattle and buffalo of different livestock farms of ten districts of Punjab province of Pakistan. Detection of CCHFV was done using enzyme link immunosorbent assay (ELISA) after proper identification of tick samples. The partial S-segment of CCHFV from ELISA positive tick samples was amplified by PCR and sequenced to determine the genotype of CCHFV. Out of2183 collected tick samples, 1913 ticks belonged to 5 species of genus Hyalomma as H. antolicum (48%), H. marginatum (30.2%), H. rufipes (10.82%), H. impressum (5.43%) and H. dromedarii (5.27%). While 270 ticks belonged to 3 species of genus Rhipicephalus as R. microplus (44.8%), R. sanguineus (32.22%) and R. turanicus (24.8%). The overall antigenic prevalence of CCHFV was found to be 12.13% in collected tick samples and 21 tick pools were sequenced for partial S-segment of CCHFV. All of the 21 tick pools were clustered in genotype IV (Asia-1). The highest prevalence of CCHFV was found in district Chakwal (24.13%) followed by Mianwali (23.68%), Rawalpindi (23.07%), Attock (20.0%), Rajanpur (10.52%) and Lahore (8.33%). In positive tick pools, the highest prevalence of CCHFV antigen was found in H. antolicum (39.6%) followed by H. marginatum (30.18%), H. rufipes (13.2%), H. impressum (3.77%), H. dromedarii (1.88%), R. microplus (5.66%) and R. sanguineus (5.66%). The current study confirms the presence of CCHFV in the ticks population of Punjab. The CCHF virus present in Punjab belongs to Asia-1 genotype. It is important to control the tick infestation of the animals present in these areas. So that the transmission cycle of CCHF can be inhibited.

Arboviruses in the Astrakhan region of Russia for 2018 season: The development of multiplex PCR assays and analysis of mosquitoes, ticks, and human blood sera.

The Astrakhan region of Russia is endemic for the number of arboviruses. In this paper, we describe the results of the detection of the list of neglected arboviruses in the Astrakhan region for the 2018 season. For the purpose of the study in-house PCR assays for detection of 18 arboviruses have been developed and validated using arboviruses obtained from Russian State Collection of Viruses. Pools of ticks (n = 463) and mosquitoes (n = 312) as well as 420 samples of human patients sera have been collected and analyzed. Using developed multiplex real-time PCR assays we were able to detect RNA of eight arboviruses (Crimean-Congo hemorrhagic fever virus, Dhori (Batken strain) virus, Batai virus, Tahyna virus, Uukuniemi virus, Inkoo virus, Sindbis virus and West Nile fever virus). All discovered viruses are capable of infecting humans causing fever and in some cases severe forms with hemorrhagic or neurologic symptoms. From PCR-positive samples, we were able to recover one isolate each of Dhori (Batken strain) virus and Crimean-Congo hemorrhagic fever virus which were further characterized by next-generation sequencing. The genomic sequences of identified Dhori (Batken strain) virus strain represent the most complete genome of Batken virus strain among previously reported.

High prevalence and different genotypes of Crimean-Congo hemorrhagic fever virus genome in questing unfed adult Hyalomma marginatum in Thrace, Turkey.

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral disease in the old continents, in many countries south of the 50 °North Parallel. The agent is known to be the most prevalent and major cause of severe and fatal human hemorrhagic diseases among the tick-borne viruses, and is the second most widespread of all medically critical arboviruses following dengue. Members of the Hyalomma genus are mainly involved in the natural transmission of the CCHF virus (CCHFV); of those, H. marginatum is known to be the primary vector of the disease in the Western Palaearctic. In general, epidemiological studies have been based on serological detections in the hosts and/or virus screening of ticks collected from the hosts. To the best of our knowledge, only a few studies have been carried out to screen the virus in unfed, questing field ticks. Nevertheless, detection of the virus in questing ticks is known to be a crucial parameter to determine the possible vector roles of the ticks and to understand the ecological dynamics of related diseases. In this study, 200 (75 males,125 females) questing H. marginatum adults collected from the field in nine villages in Thrace, located in the European part of Turkey, were screened individually for CCHFV using nested PCR. As a result, 103 (51.5 %) ticks were determined as positive with various strains of CCHFV. High positivity in questing vectors in a region where a significantly lower number of human cases have been encountered suggests that there should be some region-specific drivers that are effective in the natural dynamics of the disease. Detailed etiological and epidemiological studies are needed to reveal the possible reason for this unexpected discrepancy.

Detection of Crimean Congo haemorrhagic fever virus in North-eastern Senegal, Bokidiawé 2019.

We diagnosed a human case of Crimean Congo hemorrhagic fever (CCHF) in Bokidiawe (North-eastern Senegal), 2019. The phylogenetic analysis revealed that the isolate belongs to genotype III and is closely related to a strain reported in Mauritania in 1984 and Spain in 2016. Distribution area of CCHF in Senegal is progressively increasing.

Detection of African genotype in Hyalomma tick pools during Crimean Congo hemorrhagic fever outbreak, Rajasthan, India, 2019.

Crimean Congo hemorrhagic fever (CCHF) is a zoonotic viral disease presenting with fever and hemorrhagic manifestations in humans. After several outbreaks of CCHF being reported from Gujarat since 2011 till 2019 and from Rajasthan in 2014 and 2015, the present study reports the CCHF outbreak which was recorded from five human cases in three districts Jodhpur, Jaisalmer, and Sirohi of Rajasthan state since August 2019 till November 2019. A high percent of positivity was recorded in livestock animal samples for the CCHFV IgG antibody. CCHF virus (CCHFV) positive human blood samples and Hyalomma tick pool samples were sequenced using next-generation sequencing method. Two different M segment genotypes, encoding glycoprotein precursor, were identified from tick pools in the study: first from Asian and second from African lineage. The L gene (polymerase) and the S gene (nucleocapsid) clustered in the Asian lineage. The present study illustrates the existence of two different CCHFV lineages being circulating within the Hyalomma tick pools in the Rajasthan state, India. We also observed 3.56% amino acid changes between the death and the survived case of CCHFV in the M gene. This report also sets an alarm to enhance human, tick and livestock surveillance in other districts of Rajasthan and nearby states of India. Biosafety measures, barrier nursing along with the availability of personal protective equipment and ribavirin drug will always be a mainstay in preventing nosocomial infection for proper case management.

Genetic diversity and phylogenetic analysis of Crimean-Congo Hemorrhagic Fever viruses circulating in Pakistan during 2019.

Being an endemic country for Crimean Congo hemorrhagic fever (CCHF), this study aimed to explore the genetic diversity of CCHF virus (CCHFV) detected in Pakistan during 2019. Serum samples from patients with clinical signs of hemorrhagic fever attending tertiary care hospitals in Pakistan were tested for CCHFV RNA using real-time PCR at Department of Virology, National Institute of Health. The partial S-gene fragments were directly sequenced to determine the prevailing CCHFV genotypes and their molecular epidemiology in Pakistan. During January-December 2019, 280 samples from suspected CCHF patients were tested and 28 (10%) were found positive on real-time PCR. Positive cases were detected from 14 districts and across all four provinces of Pakistan with majority reported during August-September. The mean age of CCHFV positive patients was 37.25 years (range 5-65 years) with a high frequency in males (92.8%; n = 26) and a case fatality rate of 40.7% was observed. Phylogenetic analysis showed that S- segment of 2019 PAK CCHFV strains (n = 13) belonged to Asia-1 genotype and clustered with regional strains from Iran, Oman, and Afghanistan. We conclude that Asia-1 genotype of CCHF virus remains endemic in Pakistan. Our findings emphasize to establish a laboratory based surveillance program to monitor the disease burden and identify outbreak hotspots for effective control.

New aspects about Crimean-Congo hemorrhagic fever (CCHF) cases and associated fatality trends: A global systematic review and meta-analysis.

Crimean-Congo hemorrhagic fever (CCHF) is the most popular tick-borne disease causing by Crimean-Congo hemorrhagic fever virus (CCHFV). There are several valuable reviews considering some fields of the CCHF aspects. While there is no a systematic review about means and trends of CCHF cases and fatality rate, means and trends of CCHF cases and fatality rates of human occupations involved in CCHF. Therefore, this meta-analysis review performed to highlight and provide a global detailed of the above CCHF aspects. Among 398 collected papers, 173 papers were become this meta-analysis review. The study results confirm that an apparent increasing CCHF cases occurred through the past decades. The trends of annually and periodically CCHF cases and fatality rates were also increased. The means of annually and periodically CCHF cases and fatality rates were 57 and 432 cases, and 10 cases and 32.2 % and 49 cases and 28.8 %, respectively. The means of annually and periodically CCHF fatality rates are about one-tenth of CCHF human cases. The mean of CCHF fatality rates in Africa (22.0 %) is lower than Asia (33.5 %) and Europe (33.8 %). Among occupations involved in CCHF, agricultural (28.9 %), health-care (19.2 %) and slaughterhouse (16.7 %) workers, and farmers (13.9 %) had the maximum CCHF fatality rates in order. Based on literature review of CCHFV S-segment aspects, several clades and genotypes are reported to distribute in Africa, Asia and Europe regions. There are very wide fields to investigate the epidemiology characteristics of CCHFV clades, genotypes and their distribution in the future.

Crimean-Congo Hemorrhagic Fever Virus Genome in Tick from Migratory Bird, Italy.

We detected Crimean-Congo hemorrhagic fever virus in a Hyalomma rufipes nymph collected from a whinchat (Saxicola rubetra) on the island of Ventotene in April 2017. Partial genome sequences suggest the virus originated in Africa. Detection of the genome of this virus in Italy confirms its potential dispersion through migratory birds.

Scoping Review of Crimean-Congo Hemorrhagic Fever (CCHF) Literature and Implications of Future Research.

Crimean-Congo hemorrhagic fever (CCHF) is one of the severe forms of high-fatality hemorrhagic fever transmitted by bite of infected ticks or body fluids of infected individuals. Lack of sufficient research and endemic potential of the disease is posing serious threats to public health. The aim of this review was to explore the current status of Crimean-Congo hemorrhagic fever virus (CCHFV) related research and to identify knowledge gaps and the areas that are yet to be explored. An interpretative scoping review methodology was followed to systematically characterize the most recent literature. Literature survey was conducted using electronic databases: PubMed, Scopus, ScienceDirect and Google Scholar. This comprehensive research yielded more than 300 records, but we excluded 100 articles based on our inclusion criteria and duplicates removal. All articles (n=85) that have been published currently were discussed in this scoping review. From a total of 303 documents retrieved, 85 met the criteria. All the documents (case studies, review articles, systematic reviews, meta-analysis, case control studies, cohort studies, randomised control trials, and longitudinal studies) were included in the study. The articles mainly cover different areas such as epidemiology, prevalence, diagnosis, pathogenesis, clinical outcomes, molecular basis, phylogenetics, transmission and treatment of CCHF. Treatment and prevention related knowledge is limited; therefore, future research should focus the development of therapeutics to mitigate the increasing risk of CCHF. Priority future goal should be studies on the molecular basis and treatment of CCHFV infection because several knowledge gaps have been identified in these areas.

Survey of Crimean-Congo Hemorrhagic Fever Enzootic Focus, Spain, 2011-2015.

During 2011-2015, we conducted a Crimean-Congo hemorrhagic fever virus (CCHFV) survey in captured ticks that were feeding mainly on wild and domestic ungulates in Spain, where presence of this virus had been reported previously. We detected CCHFV RNA in Hyalomma lusitanicum and H. marginatum ticks for 3 of the 5 years. The rate of infected ticks was 2.78% (44/1,579), which was similar to those for other countries in Europe with endemic foci for CCHFV (Kosovo, Bulgaria, and Albania). These data confirm the established spread of CCHFV into western Europe. Phylogenetic study of the small RNA segment showed Africa-3 clade as the only genotype identified, although we observed cocirculation of genetic variants during 2011 and 2015. We could not rule out genetic reassortments because of lack of sequence data for the medium and large RNA segments of the virus genome.

Lassa and Crimean-Congo Hemorrhagic Fever Viruses, Mali.

We report detection of Lassa virus and Crimean-Congo hemorrhagic fever virus infections in the area of Bamako, the capital of Mali. Our investigation found 2 cases of infection with each of these viruses. These results show the potential for both of these viruses to be endemic to Mali.

Characterization of Novel Reoviruses Wad Medani Virus (Orbivirus) and Kundal Virus (Coltivirus) Collected from Hyalomma anatolicum Ticks in India during Surveillance for Crimean Congo Hemorrhagic Fever.

In 2011, ticks were collected from livestock following an outbreak of Crimean Congo hemorrhagic fever (CCHF) in Gujarat state, India. CCHF-negative Hyalomma anatolicum tick pools were passaged for virus isolation, and two virus isolates were obtained, designated Karyana virus (KARYV) and Kundal virus (KUNDV), respectively. Traditional reverse transcription-PCR (RT-PCR) identification of known viruses was unsuccessful, but a next-generation sequencing (NGS) approach identified KARYV and KUNDV as viruses in the Reoviridae family, Orbivirus and Coltivirus genera, respectively. Viral genomes were de novo assembled, yielding 10 complete segments of KARYV and 12 nearly complete segments of KUNDV. The VP1 gene of KARYV shared a most recent common ancestor with Wad Medani virus (WMV), strain Ar495, and based on nucleotide identity we demonstrate that it is a novel WMV strain. The VP1 segment of KUNDV shares a common ancestor with Colorado tick fever virus, Eyach virus, Tai Forest reovirus, and Tarumizu tick virus from the Coltivirus genus. Based on VP1, VP6, VP7, and VP12 nucleotide and amino acid identities, KUNDV is proposed to be a new species of Coltivirus Electron microscopy supported the classification of KARYV and KUNDV as reoviruses and identified replication morphology consistent with other orbi- and coltiviruses. The identification of novel tick-borne viruses carried by the CCHF vector is an important step in the characterization of their potential role in human and animal pathogenesis.IMPORTANCE Ticks and mosquitoes, as well Culicoides, can transmit viruses in the Reoviridae family. With the help of next-generation sequencing (NGS), previously unreported reoviruses such as equine encephalosis virus, Wad Medani virus (WMV), Kammavanpettai virus (KVPTV), and, with this report, KARYV and KUNDV have been discovered and characterized in India. The isolation of KUNDV and KARYV from Hyalomma anatolicum, which is a known vector for zoonotic pathogens, such as Crimean Congo hemorrhagic fever virus, Babesia, Theileria, and Anaplasma species, identifies arboviruses with the potential to transmit to humans. Characterization of KUNDV and KARYV isolated from Hyalomma ticks is critical for the development of specific serological and molecular assays that can be used to determine the association of these viruses with disease in humans and livestock.

[Results of investigation of ticks in Volga river delta (Astrakhan region, 2017) for Crimean-Congo hemorrhagic fever virus (Nairoviridae, Orthonairovirus, CCHFV) and other tick-borne arboviruses.]

INTRODUCTION: There are natural foci of Crimean-Congo hemorrhagic fever (CCHF) that vectored by Hyalomma marginatum ticks in Volga river delta (Astrakhan region, South of Russia). The circulation of Dhori virus (DHOV) (Thogotovirus: Orthomyxoviridae) has been also shown here. We hypothesized that other tick-borne arboviruses are also likely to circulate in the region. In particular, Bhanja virus (Phlebovirus: Phenuiviridae), Wad Medani virus (Orbivirus: Reoviridae), and Tamdy virus (Orthonairovirus: Nairoviridae), which were found to circulate in neighboring regions and are vectored by Haemaphysalis spp., Dermacenter spp., and Hyalomma spp. ticks. OBJECTIVES: The aim of the study was to examine ixodid ticks in Volga river delta for the presence of CCHFV, DHOV, Bhanja virus, Wad Medani virus, and Tamdy virus. MATERIAL AND METHODS: Ticks were collected in Volga river delta in 2017. We used molecular genetic methods for the detection and analysis of nucleic acids (PCR, sequencing, phylogenetic analysis). RESULTS: We detect CCHFV and DHOV RNA in H. marginatum ticks. The rate of infected H. marginatum ticks was 1.98% for CCHFV and 0.4% for DHOV. The results of genetic analysis showed that found DHOV strains are almost identical (99-100% in the M gene) and forms a separate genetic lineage alongside of Batken virus from Central Asia. At the same time, Bhanja virus, Wad Medani virus, and Tamdy virus were not found in ticks, collected in this region. CONCLUSIONS: DHOV is circulating in the natural foci of CCHF in the Volga river delta. The ratio of infection of H. marginatum with CCHFV and DHOV was determined for the first time.

Crimean-Congo Hemorrhagic Fever Virus, Mongolia, 2013-2014.

During 2013-2014, we collected 1,926 serum samples from humans and 4,583 ticks (Hyalomma asiaticum or Dermacentor nuttalli) in select regions of Mongolia to determine the risk for Crimean-Congo hemorrhagic fever virus (CCHFV) infection among humans in this country. Testing of human serum samples by ELISA demonstrated an overall CCHFV antibody prevalence of 1.4%; Bayankhongor Province had the highest prevalence, 2.63%. We pooled and analyzed tick specimens by real-time reverse transcription PCR; 1 CCHFV-positive H. asiaticum tick pool from Ömnögovi was identified. In phylogenetic analyses, the virus's partial small segment clustered with CCHFV isolates from Central Asia, and the complete medium segment grouped with CCHFV isolates from Africa, Asia, and the Middle East. This study confirms CCHFV endemicity in Mongolia and provides information on risk for CCHFV infection. Further research is needed to better define the risk for CCHFV disease to improve risk mitigation, diagnostics, and surveillance.

Viral metagenomics, genetic and evolutionary characteristics of Crimean-Congo hemorrhagic fever orthonairovirus in humans, Kosovo.

Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV) is a tick-borne virus which causes severe disease in humans with fatality cases up to 30%. We investigated the genetic and evolutionary characteristics of CCHFV in Kosovo, in particular in humans and found that different virus variants of genotype V circulate, with Turkey as a possible origin for the progenitor of southern European CCHF outbreaks. Phylogenetic analyses also revealed a single introduction event and in situ evolution of CCHFV in this country. The viral metagenomics revealed a more abundant virome in the fatal CCHF cases and the presence of a novel tick-borne segmented RNA virus belonging to the recently discovered Jingmenvirus group which raises questions about the potential pathogenic effect of this novel virus on human and animal health.

Prevalence and Phylogenetic Analysis of Crimean-Congo Hemorrhagic Fever Virus in Ticks from Different Ecosystems in Xinjiang, China.

The Crimean-Congo hemorrhagic fever virus (CCHFV), a member of the genus Orthonairovirus and family Nairoviridae, is transmitted by ticks and causes severe hemorrhagic disease in humans. To study the epidemiology of CCHFV in different ecosystems in Xinjiang, China, a total of 58,932 ticks were collected from Tarim Basin, Junggar Basin, Tianshan Mountain, and Altai Mountain from 2014 to 2017. Hyalomma asiaticum asiaticum was the dominant tick species in Tarim and Junggar basins, whereas Dermacentor nuttalli and Hyalomma detritum were found in Tianshan Mountain and Altai Mountain, respectively. Reverse transcription-polymerase chain reaction of the CCHFV small (S) genome segment was used for the molecular detection. The CCHFV-positive percentage was 5.26%, 6.85%, 1.94%, and 5.56% in Tarim Basin, Junggar Basin, Tianshan Mountain, and Altai Mountain, respectively. Sequences of the S segment were used for phylogenetic analysis and the results showed that the newly identified CCHFV strains belonged to two clades. Our study confirms that H. asiaticum asiaticum is the major vector of CCHFV in desert habitats which is consistent with previous studies, and also suggests that H. detritum and D. nuttalli are emerging vectors for CCHFV in Xinjiang. Moreover, this study reports the presence of CCHFV in the mountain habitat of Xinjiang for the first time, suggesting that future surveillance of CCHFV should also include mountainous areas.