Sheep and goats as indicator animals for the circulation of CCHFV in the environment.

Crimean--Congo hemorrhagic fever (CCHF) is a zoonotic viral disease that is asymptomatic in infected animals, but a serious threat to the health of humans.Human infections begin with non--specific febrile symptoms, but progresses to a serious haemorrhagic syndrome with a high case fatality rate. 1,2CCHF is caused by the CCHF virus (CCHFV), a member of the genus Nairovirus in the family Bunyaviridae.The virus is stable for up to 10 days in blood kept at 40°C.Although the causative virus is often transmitted by ticks, animal--to--human and human--to--human transmission also occurs.CCHF affects mostly adults (no case has been reported in children under 15 in South Africa since 1993) 3 and is endemic in many countries in Africa, Europe and Asia.During 2001, cases or outbreaks were recorded in Iran, Pakistan, South Africa with the latest being in India. 4It has also been found in parts of Europe including southern portions of the former USSR (Crimea, Astrakhan, Rostov, Uzbekistan, Kazakhstan, and Tajikistan), Turkey, Bulgaria, Greece, Albania and Kosovo (a province of the former Yugoslavia). 5--10Limited serological evidence suggests that CCHFV might also occur in parts of Hungary, France and Portugal.The most recent CCHF outbreak occurred in January 2011 in the state of Ahmedabad (India).In this outbreak this rare deadly virus killed three people.The three victims included an adult female, a nurse, and the doctor who treated the adult female at a private hospital in Ahmedabad (India). 11The patients died due to multiple organ failure, specifically failure of the liver and kidney.The National Institute of Virology (NIV), (Pune, India) confirmed that all three patients were infected with the CCHF virus.11 The NIV is testing some 50 samples from the area, and the Gujarat government, warning of a possible outbreak, has begun a screening exercise covering approximately 16,000 villagers.No CCHF case has ever been reported in India before.

A one-step multiplex real-time RT-PCR for the universal detection of all currently known CCHFV genotypes

Unique strain of Crimean-Congo hemorrhagic fever virus, Mali.

BackgroundThere are only few assays available for the detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)-specific antibodies in animals, and data about diagnostic sensitivity and specificity are incompletely documented for most of these tests. This is unfortunate since CCHFV antibodies in animals can be used as indicator for virus circulation in a geographic area and therewith potential risk of human exposure. This paper therefore reports on a novel ELISA for the detection of CCHFV-specific antibodies in cattle and on its application for testing ruminant sera from the Former Yugoslav Republic of Macedonia.Principal FindingsA highly sensitive and specific ELISA was developed to detect CCHFV-specific IgG antibodies in cattle. The assay was validated by using 503 negative serum samples from a country where CCHFV has never been detected until now, and by using 54 positive serum samples. The positive sera were verified by using two commercially available assays (for testing human serum) which we have adapted for use in animals. The sensitivity of the novel ELISA was 98% and its specificity 99%. The presence of Hyalomma ticks was demonstrated in the Former Yugoslav Republic of Macedonia and depending on the region antibody prevalence rates up to 80% were detected in the cattle population.ConclusionThis article describes a fully validated, highly sensitive and specific ELISA for the detection of CCHFV-specific IgG antibodies in cattle. Using this assay, CCHFV-specific antibodies were detected for the first time in cattle in the Former Yugoslav Republic of Macedonia, giving evidence for an active circulation of this virus in the country. Supporting this conclusion, the occurrence of the main vector of CCHFV was demonstrated in the present work for the first time in Former Yugoslav Republic of Macedonia.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus, and zoonosis, and affects large regions of Asia, Southwestern and Southeastern Europe, and Africa. CCHFV can produce symptoms, including no specific clinical symptoms, mild to severe clinical symptoms, or deadly infections. Virus isolation attempts, antigen-capture enzyme-linked immunosorbent assay (ELISA), and reverse transcription polymerase chain reaction (RT-PCR) are all possible diagnostic tests for CCHFV. Furthermore, an efficient, quick, and cheap technology, including biosensors, must be designed and developed to detect CCHFV. The goal of this article is to offer an overview of modern laboratory tests available as well as other innovative detection methods such as biosensors for CCHFV, as well as the benefits and limits of the assays. Furthermore, confirmed cases of CCHF are managed with symptomatic assistance and general supportive care. This study examined the various treatment modalities, as well as their respective limitations and developments, including immunotherapy and antivirals. Recent biotechnology advancements and the availability of suitable animal models have accelerated the development of CCHF vaccines by a substantial margin. We examined a range of potential vaccines for CCHF in this research, comprising nucleic acid, viral particles, inactivated, and multi-epitope vaccines, as well as the present obstacles and developments in this field. Thus, the purpose of this review is to present a comprehensive summary of the endeavors dedicated to advancing various diagnostic, therapeutic, and preventive strategies for CCHF infection in anticipation of forthcoming hazards.Graphical For the protection of medical personnel and effective case management, an early diagnosis of Crimean-Congo hemorrhagic fever (CCHF) is critical. CCHF is diagnosed through laboratory procedures such as RT-PCR, ELISA, virus isolation attempts, and ELISA detection of IgG and IgM antibodies. This review examines several biomarkers researched for their potential use in the diagnosis and prognosis of critical viral infections. It also explores the utility of more traditional diagnostic markers in predicting secondary complications, distinguishing Crimean-Congo hemorrhagic fever virus (CCHFV) infection, and serving as severity indicators. CCHFV vaccine development is advancing at an accelerated rate, facilitated by the availability of a lethal animal infection model. Hence, this review aims to furnish a comprehensive synopsis of the endeavors devoted to various vaccine candidates utilizing distinct approaches against CCHFV. These candidates comprise inactivated, virus-like particles, recombinant proteins, and nucleic acid vaccines. Furthermore, supportive therapy serves as the principal modality of treatment. Human cases of CCHF have been treated with ribavirin, a broad-spectrum antiviral medication; nevertheless, the therapeutic advantages of this intervention remain elusive. This article analyzes the present advancements and prospective trajectories in the realm of antiviral approaches targeting CCHFV.

The Crimean Congo Haemorrhagic Fever (CCHF) is a viral infection caused by the CCHF virus (CCHFV), a member of the family Bunyaviridae, genus Nairovirus. This virus is transmitted by ticks of the genus Hyalomma and has been reported in more than 30 countries in Africa, Asia, South-East Europe and the Middle East [1] arising considerable public health concern in many regions of the world. The first case of CCHF was probably described by a Physician in Tajikistan in 1110 DC in a patient with hemorrhagic manifestations [2]. More recently, CCHF was described for the first time as a proper clinical entity in 1944-45 when about 200 Soviet soldiers were infected while providing assistance to farmers in the Crimea during the Second World War [3]. However the virus was first isolated from a Patient in the Democratic Republic of the Congo only in 1956 [4]. The disease is currently reported in Asia (China, Kazakhstan, Tajikistan, Pakistan, Afghanistan, Iraq, Iran, UAE, Oman, Yemen, southern Saudi Arabia), Africa (Mauritania, Senegal, Burkina Faso, Congo, Uganda, Kenya, Tanzania, Namibia, South Africa) [5] and Europe [6]. H. marginatum, the main vector of CCHF virus in Europe, is located in Albania, Bulgaria, Cyprus, France, Greece, Italy, Kosovo, Moldova, Portugal, Romania, Russia, Serbia, Spain, Turkey and Ukraine. In 2006 it was isolated for the first time in the Netherlands and in southern Germany [7,8]. As evidenced by a recent study (Estrada-Pena, 2012), favorable climate and ecological conditions present in several European countries bordering the Mediterranean Sea, could ensure a CCHFV expansion in the near future. The risk of infection is particularly high in Italy due to the presence of concomitant factors such as the wild ecological heterogeneity that characterizes its territory, and the peculiar position in the middle of the Mediterranean area. Besides, the recent raising of the average seasonal temperatures in south European countries could increase the colonization by vector populations that were not naive [9]. Typically CCHF’s clinical manifestations are high fever, malaise, severe headache and gastrointestinal symptoms. Extensive bleeding may occur in an advanced stage of the disease, with mortality rates ranging from 5% to 50%. CCHF is a disease of immediate communication to the public health authorities [4]. During the last decade the CCHFV has emerged and / or re-emerged in several countries of the Balkans, Turkey, in the regions of the south-west of the Russian Federation and Ukraine, with a significantly high mortality rates. The reasons for the re-emergence of CCHFV include climate and anthropogenic factors such as changes in agricultural practices or hunting activities and cattle movement. These factors can influence the dynamics of host-tick-virus. In the light of the dangerousness of this disease an International surveillance for the management of an outbreak of CCHF is needed. This should be carry out with an integrated approach among the countries at risk of infection in the Mediterranean area in order to design measures for prevention and control of this disease, mapping of endemic areas and assessing risk of CCHF transmission. In addition, the areas at risk of further expansion of CCHF should be identified also oversight [6]. Different elements concerning diagnosis, monitoring and treatment of CCHF should be considered in order to increase the surveillance in Europe defining appropriate measures for prevention and control. Currently there are no standard definitions regarding how to notify CCHF infection and how “contact tracing” must be carry out in different European countries. Recent cases of nosocomial infection of health care workers infected by CCHFV have been documented. These cases stress the needing for a focused education, with particular regard to transmission pathways and to preventive measures; it is also crucial providing adequate resources on contrasting the diffusion of this disease [6]. Besides, in many countries at risk of infection, standard collaboration procedures and protocols for data exchange are not settled between human and animal health services. Therefore designing and creating a surveillance system for these vector-transmitted diseases should be aimed to develop common procedures and protocols for data exchange. References

Seroprevalence of Crimean-Congo Hemorrhagic Fever Virus, Bulgaria

Tick-borne diseases pose a serious threat to human health in South-Eastern Europe, including Kosovo. While Crimean–Congo hemorrhagic fever (CCHF) is a well-known emerging infection in this area, there are no accurate data on Lyme borreliosis and tick-borne encephalitis (TBE). Therefore, we sampled and tested 795 ticks. Ixodes ricinus (n = 218), Dermacentor marginatus (n = 98), and Haemaphysalis spp. (n = 24) were collected from the environment by flagging (all from Kosovo), while Hyalomma marginatum (n = 199 from Kosovo, all from Kosovo) and Rhipicephalus bursa (n = 130, 126 from Albania) could be collected only by removal from animal pasture and domestic ruminants. Ticks were collected in the years 2014/2015 and tested for viral RNA of CCHF and TBE viruses, as well as for DNA of Borrelia burgdorferi sensu lato by real-time PCR. In Kosovo, nine ticks were positive for RNA of Crimean–Congo hemorrhagic fever virus and seven for DNA of B. burgdorferi s. l. None of the ticks tested positive for TBEV. CCHF virus was detected in one H. marginatum male specimen collected while feeding on grazing cattle from the Prizren region and in eight R. bursa specimens (five females and three males collected while feeding on grazing sheep and cattle) from the Prishtina region (Kosovo). B. burgdorferi s. l. was detected in seven questing ticks (four male and one female D. marginatus, two I. ricinus one female and one male) from the Mitrovica region (Kosovo). Our study confirmed that CCHF virus is circulating in Kosovo mainly in H. marginatum and R. bursa in the central areas of the country. B. burgdorferi s. l. was found in its major European host tick, I. ricinus, but also in D. marginatus, in the north of the Kosovo. In order to prevent the spread of these diseases and better control of the tick-borne infections, an improved vector surveillance and testing of ticks for the presence of pathogens needs to be established.

Epidemiological investigations of Crimean-Congo haemorrhagic fever virus infection in sheep and goats in Balochistan, Pakistan

Molecular phylogenetic study on S-segment of Crimean–Congo Hemorrhagic Fever (CCHF) virus genome isolated from ticks in in West Azerbaiajan and Sistan and Baluchestan provinces, Iran, 2010

A novel double-antigen sandwich ELISA for the species-independent detection of Crimean-Congo hemorrhagic fever virus-specific antibodies

To the Editor: Crimean-Congo hemorrhagic fever virus (CCHFV) (Nairovirus, Bunyaviridae), the causative agent of Crimean-Congo hemorrhagic fever, has been detected in sub-Saharan Africa, southeastern Europe, the Middle East, and central Asia. The virus has been detected in >31 species of ticks and is transmitted to humans by bite of infected ticks (mainly of the genus Hyalomma) or by contact with body fluids or tissue of viremic patients or livestock. The disease is characterized by fever, myalgia, headache, vomiting, and sometimes hemorrhage; reported mortality rate is 10%–50% (1). CCHFV strains currently constitute 7 evolutionary lineages, 1 of which (Europe 2) contains the prototype strain AP92, which was isolated in 1975 from Rhipicephalus bursa ticks collected from goats in Greece (2). This strain seems to have low or no pathogenicity for humans; only a few mild cases have been reported (3). This observation is supported by the relatively high (14.4%) seroprevalence but no clinical cases in humans in northwestern Greece (4). The documented tick carriers of this strain are R. bursa and Hyalomma marginatum (5). Hyalomma aegyptium ticks are highly host specific; adults feed almost entirely on tortoises of the genus Testudo (6) and occassionally on hedgehogs and hares. Unlike adult ticks, the larvae and nymphs are less host specific and feed on a wide spectrum of hosts (e.g., other reptiles, birds, and mammals [including humans]) (7). This trait elevates the epidemiologic role of the tick as a possible bridge vector connecting wildlife, domestic animals, and humans. To determine the biological and epidemiological role of H. aegyptium ticks, during 2009–2010, we collected 56 adult ticks from 12 Testudo graeca tortoises at a locality near the city of Aflou in Laghouat Province, Algeria. We tested the ticks for probable CCHFV infection by using nested reverse transcription PCR (8), which amplifies a partial fragment of the CCHFV small RNA segment. We slightly modified the assay: reverse transcription time was 60 minutes and annealing temperature was 52°C (9). In total, 16 (28.6%) ticks were positive for CCHFV. The PCR products of 15 (26.8%) positive samples were sequenced. BLAST (http://blast.ncbi.nlm.nih.gov//Blast.cgi) analysis identified all 15 sequences as CCHFV with 98%–100% identity to the AP92 strain (GenBank accession no. {type:entrez-nucleotide,attrs:{text:DQ211638,term_id:78191750,term_text:DQ211638}}DQ211638). Two variants of AP92 were detected and differed by 0.6%. A phylogenetic tree was constructed by Bayesian inference, using MrBayes version 3.1.2 (http://mrbayes.sourceforge.net/index/php) under a general time-reversible plus gamma distribution plus invariable site model with 107 generations setup (Figure). Sequences are available in GenBank (accession nos. KT99097 and KT99098). Figure Phylogenetic analysis of Crimean-Congo hemorrhagic fever virus small RNA segment sequences, performed by using Bayesian inference in MrBayes version 3.1.2. (http://mrbayes.csit.fsu.edu/) under a general time-reversible plus gamma distribution plus invariable ... Our findings demonstrate the presence of CCHFV in Algeria, either recently introduced or overlooked. The nearest location where CCHFV has been reported is the Zouala region in Morocco, where the virus was detected in H. marginatum tick larvae and nymphs collected from migratory birds (10). It also confirms association of AP92-like sequences with H. aegyptium ticks. This study shows that the Europe 2 lineage is not restricted to the Balkan region and Turkey. The role of H. aegyptium ticks as CCHFV vectors should be further tested. Further investigation of the distribution of CCHFV in ticks in Algeria is also needed. To date, CCHFV strains of lineage Europe 2 have not been associated with severe disease in humans. However, physicians in Algeria should be aware of potential Crimean-Congo hemorrhagic fever cases.

Crimean-Congo hemorrhagic fever (CCHF) virus is transmitted to humans by Hyalomma ticks or by direct contact with the blood of infected humans or domestic animals. CCHF virus has been reported from the Near, Middle, and Far East (countries such as Iraq, Pakistan, United Arab Emirates, Kuwait, Oman, and China [1, 6-8, 11]) and from several African countries (4, 9). Besides, there are several reports on CCHF virus in the former Yugoslavia (5, 10), but CCHF virus strains from this area have not been characterized up to now. We describe here a case of CCHF in the year 2000 in Kosovo that preceeded an outbreak in the same region in 2001 (2). On 23 May 2000, a farmer's wife (age, 17 years) living near Pristina, Kosovo, was bitten by a tick in the left femoral region while working in her garden. The tick was later identified as a member of the Hyalomma species at our institute. The disease started on 28 May with chills, myalgia, nausea, anorexia, vomiting, headache, and backache. The victim visited an outpatient clinic in Prizren, where broad-range antibiotic therapy was initiated because a septic infection could not be excluded. On 30 May, she developed massive hemorrhage with hematemesis (7 to 8 times per day), melena, hematuria, metrorrhagia, and petechia. She was hospitalized in a medical ward with no special isolation measures in a difficult clinical condition on 31 May, without palpable pulse or measurable blood pressure, with a body temperature of 39.7°C, and with severe hemorrhagic manifestations (petechiae, melena, hematemesis, and metrorrhagia). On 1 June, epistaxis and gingival bleeding were additionally observed and the high fever continued (40.1°C). The patient was fully orientated, without neurological symptoms, but prostrate. She complained of back pain and headache. Her eyes showed signs of hemorrhagic conjunctivitis and her gingiva and tongue were covered with hemorrhagic crusts, but the petechiae in the pectoral region were already in remission. Large ecchymoses appeared at the sites of venipuncture. The heart had a sinusal rhythm (90 beats/min) and weak tones, without noise. The abdomen was tender on palpation and the liver was palpable and slightly enlarged, but the spleen was not palpable. Diuresis was evident, with 1,500 ml of urine excreted per day. On 3 June she developed a polyuria (4,500 ml per day). The next day, the hemorrhagic diathesis had almost disappeared with only a light residual metrorrhagia. Blood pressure was 100/70 mmHg (pulse frequency, 60 beats/min). She was feeling better. Supportive therapy given to the patient during the course of the disease consisted of hydration and control of temperature. No blood transfusions were administered. No secondary cases have occurred in the hospital where the patient was treated.

Crimean-Congo hemorrhagic fever (CCHF) is a viral disease transmitted to humans mainly by bite of Ixodid ticks, mainly those of the Hyalomma genus. CCHFV belongs to the genus Nairovirus in the family Bunyaviridae. CCHF virus is a segmented, single stranded, negative sense and RNA viruses. The onset of the disease is very sudden, with symptoms such as fever, rigors, intense headache, chills, and backache or leg pains, myalgia, nausea, and vomiting. CCHF originally identified in the former Soviet Union and the Congo, has rapidly spread across large sections of Europe, Asia, and Africa, and has been reported in more than 30 countries. The climatic changes may affect the life cycle of ticks and the routes of migratory birds, leading to tick abundance and virus distribution in CCHF-free areas. Extended use of land for agriculture and farming and changes in hunting activities play also a role in CCHF incidence, while livestock trade and movement may influence host-tick-virus dynamics resulting in transfer of CCHFV-infected ticks in non-endemic areas. Recent years, the epidemiology of CCHF is changing in Balkans and Turkey. Balkan Peninsula is a known endemic CCHF area, and sporadic cases and even outbreaks are being reported every year. The annual number of human CCHF cases is increasing in Balkans and Turkey. While Bulgaria, Kosovo and Albania were ZET Krm-Kongo Kanamal Atei (KKKA), zellikle Ixodid cinsi kene sr (esas olarak Hyalomma cinsi)

Crimean-Congo haemorrhagic fever virus (CCHFV) and Nairobi sheep disease virus (NSDV) are orthonairoviruses of concern, able to cause haemorragic fever disease in humans and sheep, respectively. CCHFV and NSDV cocirculating in small ruminant populations across South Asia and East Africa. Cross-reactivity to viruses of the Orthonairovirus genus can potentially interfere with serological assays when employed for serosurveillance in regions where two or more genus members overlap in their distribution. In this study, sheep sera sampled from a region of confirmed CCHFV circulation and NSDV absence were utilized, thereby eliminating the possibility of co-exposure. Field sera were tested against in-house anti-NSDV ELISAs specific to the nucleoprotein (NSDV NP) and glycoprotein C (NSDV Gc) antigens as well as an in-house NSDV 80% plaque reduction neutralization test (PRNT80). We assessed whether there is a correlation between CCHFV- and NSDV-specific ELISAs. Furthermore, epitopes-derived from CCHFV antigens for sheep antibody that were available from the literature were analyzed. When comparing NSDV antigen-specific antibody responses against previously tested CCHFV antigen-specific antibody responses, a strong positive correlation was observed between the Gc-specific responses, while a weak positive correlation was observed between the NP-specific responses. Consequently, NP-specific ELISAs have a higher assay specificity compared to Gc-specific ELISAs, making them more suitable for serosurveillance in regions where multiple orthonairoviruses co-circulate. Crucially, only one seropositive sample to NSDV Gc-specific out of a set of 224 (0.4%) showed a neutralizing capacity at the lowest serum dilution (1:8), suggesting these field sera have not been exposed to NSDV. Based on an analysis of known epitopes in NP targeted by antibodies in sheep serum, we propose that NP is less cross-reactive because dominant epitopes are highly dissimilar between CCHFV and NSDV. Gc exhibited a strong cross-reaction while the NP was weakly cross-reactive due to dominant epitopes being highly dissimilar between CCHFV and NSDV. Our in-house PRNT80 assay can could be used as a confirmatory test in regions where CCHFV and NSDV circulate.