The DEVD motif of Crimean- Congo hemorrhagic fever virus nucleoprotein is essential for viral replication in tick cells

The DEVD motif of Crimean-Congo hemorrhagic fever virus nucleoprotein is essential for viral replication in tick cell

Molecular identification of adenoviruses associated with respiratory infection in Egypt from 2003 to 2010.

BACKGROUND: Human adenoviruses of species B, C, and E (HAdV-B, -C, -E) are frequent causative agents of acute respiratory infections worldwide. As part of a surveillance program aimed at identifying the etiology of influenza-like illness (ILI) in Egypt, we characterized 105 adenovirus isolates from clinical samples collected between 2003 and 2010. METHODS: Identification of the isolates as HAdV was accomplished by an immunofluorescence assay (IFA) and confirmed by a set of species and type specific polymerase chain reactions (PCR). RESULTS: Of the 105 isolates, 42% were identified as belonging to HAdV-B, 60% as HAdV-C, and 1% as HAdV-E. We identified a total of six co-infections by PCR, of which five were HAdV-B/HAdV-C co-infections, and one was a co-infection of two HAdV-C types: HAdV-5/HAdV-6. Molecular typing by PCR enabled the identification of eight genotypes of human adenoviruses; HAdV-3 (n = 22), HAdV-7 (n = 14), HAdV-11 (n = 8), HAdV-1 (n = 22), HAdV-2 (20), HAdV-5 (n = 15), HAdV-6 (n = 3) and HAdV-4 (n = 1). The most abundant species in the characterized collection of isolates was HAdV-C, which is concordant with existing data for worldwide epidemiology of HAdV respiratory infections. CONCLUSIONS: We identified three species, HAdV-B, -C and -E, among patients with ILI over the course of 7 years in Egypt, with at least eight diverse types circulating

Crimean-Congo Hemorrhagic Fever Virus in High-Risk Population, Turkey

In the Tokat and Sivas provinces of Turkey, the overall Crimean-Congo hemorrhagic fever virus (CCHFV) seroprevalence was 12.8% among 782 members of a high-risk population. CCHFV seroprevalence was associated with history of tick bite or tick removal from animals, employment in animal husbandry or farming, and being >40 years of age

Nosocomial Outbreak of Crimean-Congo Hemorrhagic Fever, Sudan

To confirm the presence of Crimean-Congo hemorrhagic fever in Sudan, we tested serum of 8 patients with hemorrhagic fever in a rural hospital in 2008. Reverse transcription–PCR identified Crimean-Congo hemorrhagic fever virus. Its identification as group III lineage indicated links to virus strains from South Africa, Mauritania, and Nigeria

Evidence of Prolonged Crimean-Congo Hemorrhagic Fever Virus Endemicity by Retrospective Serosurvey, Eastern Spain

We conducted a retrospective serosurvey for antibodies against Crimean-Congo hemorrhagic fever virus in wild ungulates along the eastern Mediterranean Coast of Spain. The virus has been endemic in this region since 2010 but is mainly restricted to geographic clusters with extremely high seropositivity associated with high density of bovids.info:eu-repo/semantics/publishedVersio

Antiviral Res

Early in the COVID-19 pandemic, demand for N95 respirators far exceeded the supply, leading to widespread shortages. Initially, the US Centers for Disease Control and Prevention did not recommend N95 respirators in nonhealthcare settings, in order to reserve them for healthcare workers. As N95s became more available, the recommendations were updated in May 2021 to include N95 respirators for nonhealthcare settings. In this study, we estimated the numbers of N95s needed for nonhealthcare essential workers in the United States. This information is valuable for crisis preparedness and planning for future large-scale communicable respiratory infectious disease epidemics or pandemics. We adapted a spreadsheet-based tool originally built to estimate the potential demand for N95 respirators during an influenza pandemic. We defined nonhealthcare essential occupations according to the 2020 US Department of Homeland Security guidance and used US Bureau of Labor Statistics employment numbers and Occupational Information Network data as model parameters. We modeled minimum, intermediate, and maximum N95 provision scenarios (as 1, 2, and 5 N95 respirators, respectively) per week per worker, for pandemic durations of 15 and 40 weeks. For 85.15 million nonhealthcare essential workers during a 15-week pandemic, an estimated 1.3 billion N95 respirators would be needed under minimum provision scenarios, 2.6 billion for intermediate provision, and 6.4 billion for maximum provision. During a 40-week pandemic, these estimates increased to 3.4 billion, 6.8 billion, and 17 billion. Public health authorities and policymakers can use these estimates when considering workplace respirator-wearing practices, including prioritization of allocation, for nonhealthcare essential workers. Our novel spreadsheet-based tool can also be used to quickly generate estimates of other preparedness and response equipment.CC999999/ImCDC/Intramural CDC HHSUnited States

Characterization of the Hazara Virus and Tick cell lines interactions towards Crimean-Congo Haemorrhagic Fever Virus control

Tomaz, F., Antunes, S., Parreira, R., Domingos, A., Seixas, G.,” Characterization of Hazara virus infection in Hyalomma lusitanicum tick cell line.” Poster ID: 185. National Congress of Microbiology and Biotechnology-MicroBiotec2. 23rd – 26th November 2021 (Online Web-conference)Crimean-Congo haemorrhagic fever virus (CCHFV) (Nairoviridae, Orthonairovirus) is a tick-borne virus that causes a severe disease with high mortality rates in humans and is classified as a biosafety level (BSL) 4 pathogen. This virus is mainly transmitted by Hyalomma spp. ticks and has been increasingly detected in Europe and other CCHFV-endemic areas. Since H. lusitanicum geographical distribution encompasses the Iberian Peninsula, where recent human cases of CCHFV disease have been detected, a more comprehensive understanding of the tick-virus interactions is needed to fully realize the role this vector plays in the (re)emergence of CCHFV. As showed in previous studies, Hazara virus (HAZV) is a valid surrogate model to study CCHFV under BSL2 conditions. The aim of this thesis was to establish and characterize the infection of three tick-derived cell lines - HLE/LULS42, HAECTVM9 and ISE6 - with HAZV to elucidate the dynamics of the infection (in vitro) in the vector. Tick cell lines were maintained in parallel with HAZV grown in Vero E6 and SW-13 cells. The three tick cell lines were infected with HAZV at different multiplicities of infection and had their supernatants retrieved at several time-points post infection and later processed for RNA extraction using tri-reagent. Viral RNA was quantified using an in-house designed quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) using a TaqMan probe targeting a region of the S segment of HAZV genome. Results obtained demonstrate that a persistent infection is formed in all three cell lines, without hampering the fitness of the tick cells. The viral titres obtained by RT-qPCR provided an image of the kinetics of the infection and show a peak of viral genomes at approximately 7-8 days post-infection. The data also shows that HLE/LULS42 – HAZV infected cells produce higher values of viral titre in all the three cell lines studied, displaying a difference in the kinetics of viral replication. Future studies are warranted in HLE/LULS42 tick cell line to study the molecular mechanisms underlying the viral persistence without damaging the host. Altogether, this work provides critical knowledge in understanding the vector-pathogen interactions and will guide future studies towards prevention and treatment of poorly studied viral threats.O vírus da febre hemorrágica da Crimeia-Congo (CCHFV) (Nairoviridae, Orthonairovirus) é um vírus transmitido por carraças responsável por uma grave doença com elevada taxa de mortalidade em humanos e é classificado como agente patogénico de nível de biossegurança (BSL) 4. Este vírus é transmitido principalmente por carraças do género Hyalomma spp. e a sua deteção na Europa e outras áreas endémicas de CCHFV tem aumentado. Visto que, a distribuição geográfica de H. lusitanicum abrange a Península Ibérica, onde foram detectados casos recentes de infeção por CCHFV, é necessária uma compreensão mais detalhada das interações entre a carraça e o vírus para melhor compreender o papel que este vetor desempenha na (re)emergência de CCHFV. Estudos prévios com o vírus Hazara (HAZV), validaram este vírus como um modelo para estudar CCHFV em condições BSL2. Esta tese teve como objectivos, estabelecer e caracterizar a infecção por HAZ, em três linhagens celulares distintas de carraças - HLE/LULS42, HAECTVM9 e ISE6 – de modo a elucidar a dinâmica de infecção (in vitro) neste vetor. As linhas celulares de carraça foram mantidas, em paralelo, com células Vero E6 e SW-13. As três linhagens celulares de carraça foram infectadas com HAZV em diferentes multiplicidades de infecção e o seu sobrenadante foi recuperado em vários pontos temporais pós-infecção, sendo posteriormente processado para extração de RNA usando tri-reagente. O RNA viral foi quantificado por meio de uma nova reação em cadeia da polimerase via transcriptase reversa em tempo real (RT-QPCR), usando uma sonda TaqMan, complementar a sequencia específica do segmento S do genoma de HAZV. Os resultados obtidos, evidenciam a formação de uma infecção persistente, presente em todas as três linhagens celulares, sem prejudicar a sobrevivência das mesmas. Os títulos virais obtidos através de RT-QPCR revelam um pico na deteção de genomas virais, aproximadamente 7-8 dias após a infecção. Estes resultados demonstram igualmente que a linhagem celular HLE/LULS42, infetada com HAZV apresenta os valores de título viral mais elevado em todas as três linhagens celulares estudadas, indicando uma diferença na cinética de replicação viral. É necessário realizar estudos futuros, usando a linha celular HLE/LULS42, para uma melhor compreensão dos mecanismos moleculares implícitos na manutenção da persistência da infeção viral sem danificar o hospedeiro. Concluindo, este trabalho fornece novos dados para a compreensão das interações entre vetor e agente patogénico e permitirá realizar novo estudos para a prevenção e tratamento de ameaças virais

Crimean-Congo hemorrhagic fever: epidemiological trends and controversies in treatment

Crimean-Congo hemorrhagic fever (CCHF) virus has the widest geographic range of all tick-borne viruses and is endemic in more than 30 countries in Eurasia and Africa. Over the past decade, new foci have emerged or re-emerged in the Balkans and neighboring areas. Here we discuss the factors influencing CCHF incidence and focus on the main issue of the use of ribavirin for treating this infection. Given the dynamics of CCHF emergence in the past decade, development of new anti-viral drugs and a vaccine is urgently needed to treat and prevent this acute, life-threatening disease

The complete genome sequence of a Crimean-Congo Hemorrhagic Fever virus isolated from an endemic region in Kosovo

The Balkan region and Kosovo in particular, is a well-known Crimean-Congo hemorrhagic fever (CCHF) endemic region, with frequent epidemic outbreaks and sporadic cases occurring with a hospitalized case fatality of approximately 30%. Recent analysis of complete genome sequences of diverse CCHF virus strains showed that the genome plasticity of the virus is surprisingly high for an arthropod-borne virus. High levels of nucleotide and amino acid differences, frequent RNA segment reassortment and even RNA recombination have been recently described. This diversity illustrates the need to determine the complete genome sequence of CCHF virus representatives of all geographically distinct endemic areas, particularly in light of the high pathogenicity of the virus and its listing as a potential bioterrorism threat. Here we describe the first complete CCHF virus genome sequence of a virus (strain Kosova Hoti) isolated from a hemorrhagic fever case in the Balkans. This virus strain was isolated from a fatal CCHF case, and passaged only twice on Vero E6 cells prior to sequence analysis. The virus total genome was found to be 19.2 kb in length, consisting of a 1672 nucleotide (nt) S segment, a 5364 nt M segment and a 12150 nt L segment. Phylogenetic analysis of CCHF virus complete genomes placed the Kosova Hoti strain in the Europe/Turkey group, with highest similarity seen with Russian isolates. The virus M segments are the most diverse with up to 31 and 27% differences seen at the nt and amino acid levels, and even 1.9% amino acid difference found between the Kosova Hoti and another strain from Kosovo (9553-01). This suggests that distinct virus strains can coexist in highly endemic areas