RESUMEN
During the summer of 2023, the European Region experienced a limited resurgence of mpox cases following the substantial outbreak in 2022. This increase was characterised by asynchronous and bimodal increases, with countries experiencing peaks at different times. The demographic profile of cases during the resurgence was largely consistent with those reported previously. All available sequences from the European Region belonged to clade IIb. Sustained efforts are crucial to control and eventually eliminate mpox in the European Region.
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Brotes de Enfermedades , Filogenia , Humanos , Europa (Continente)/epidemiología , Masculino , Femenino , Adulto , Persona de Mediana Edad , Adolescente , Adulto Joven , Niño , Anciano , Vigilancia de la Población , Preescolar , IncidenciaRESUMEN
Social distancing, mask-wearing, and travel restrictions during the COVID-19 pandemic have significantly impacted the spread of influenza viruses. The objectives of this study were to analyze the pattern of influenza virus circulation with respect to that of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Bulgaria during the 2021-2022 season and to perform a phylogenetic/molecular analysis of the hemagglutinin (HA) and neuraminidase (NA) sequences of representative influenza strains. Influenza infection was confirmed using real-time reverse transcription polymerase chain reaction in 93 (4.2%) of the 2193 patients with acute respiratory illness tested wherein all detected viruses were subtyped as A(H3N2). SARS-CoV-2 was identified in 377 (24.3%) of the 1552 patients tested. Significant differences in the incidence of influenza viruses and SARS-CoV-2 were found between individual age groups, outpatients/inpatients, and in the seasonal distribution of cases. Two cases of coinfections were identified. In hospitalized patients, the Ct values of influenza viruses at admission were lower in adults aged ≥65 years (indicating higher viral load) than in children aged 0-14 years (p < 0.05). In SARS-CoV-2-positive inpatients, this association was not statistically significant. HA genes of all A(H3N2) viruses analyzed belonged to subclade 3C.2a1b.2a. The sequenced viruses carried 11 substitutions in HA and 5 in NA, in comparison to the vaccine virus A/Cambodia/e0826360/2020, including several substitutions in the HA antigenic sites B and C. This study revealed extensive changes in the typical epidemiology of influenza infection, including a dramatic reduction in the number of cases, diminished genetic diversity of circulating viruses, changes in age, and seasonal distribution of cases.
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COVID-19 , Vacunas contra la Influenza , Gripe Humana , Adulto , Niño , Humanos , Subtipo H3N2 del Virus de la Influenza A/genética , SARS-CoV-2/genética , Estaciones del Año , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Bulgaria/epidemiología , Filogenia , Prevalencia , Pandemias , COVID-19/epidemiología , ARN Viral/genética , Análisis de Secuencia de ADN , Hemaglutininas , Neuraminidasa/genéticaRESUMEN
Installing efficient protective immunity by anti-SARS-CoV-2 vaccines is the only current means to overcome coronavirus disease 2019 pandemics. The cellular and humoral immune responses induced with an messenger RNA (mRNA) (BNT162b2) or with a vector (ChAdOx1nCoV-19) vaccine among Bulgarian healthcare workers (n = 123, aged 23-71 years) were studied in the course of 16 weeks after priming. Receptor-binding domain (RBD)-blocking Abs and SARS-CoV-2 RBD immunoglobulin A (IgA) were evaluated in parallel with interferon gamma (IFNγ)-producing virus-specific T cells. Both vaccines induced RBD-blocking Abs in 100% of the participants after complete immunization while the levels of protection after a single dose largely varied (22%-98%). Advanced age had a negative impact on the level and longevity of virus-neutralizing activity induced by one dose mRNA, but not by the vector vaccine. RBD-binding IgA was detected in 100% of tested donors from the mRNA vaccine cohort, and in 67% of tested from the vector vaccine cohort, at least 1 month after completed immunization. One month after completing mRNA immunization, the number of IFNγ-producing T cells correlated significantly with the levels of RBD-specific IgA and virus-neutralizing activity induced after priming. Enumeration of circulating virus-specific IFNγ+ T cells is not recommended for evaluation of protective immunity as their detection may require longer stimulation beyond the firstmonth postimmunization. In conclusion, BNT162B2 and ChAdOx1nCoV-19 induced potent and comparable humoral and cellular anti-SARS-CoV-2 immune responses, peaking between 10 and 30 days after complete immunization. A single dose of any vaccine did not induce adequate protection in a great part of donors, making the shorter interval between mRNA vaccine doses preferable in the settings of increased risk of infection.
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COVID-19 , Adulto , Anciano , Anticuerpos Antivirales , Vacuna BNT162 , Bulgaria , COVID-19/prevención & control , Estudios de Cohortes , Personal de Salud , Humanos , Inmunidad Celular , Inmunidad Humoral , Persona de Mediana Edad , Estudios Prospectivos , ARN Mensajero , SARS-CoV-2 , Vacunas Sintéticas , Adulto Joven , Vacunas de ARNmRESUMEN
The evolution of the emerging SARS-CoV-2 variants carrying mutations in the spike protein raises concerns about the possibility of accelerated transmission in the ever-evolving COVID-19 pandemic worldwide. AY.4.2, a sublineage of the Delta variant, was considered a variant under investigation (VUI) and also gained the nickname "Delta Plus," due to its extra mutations, Y145H and A222V. In this study, using genomic epidemiology, we provide the first insights into the introduction of AY.4.2 in Bulgaria and the AY.4.2.1 sublineage that found larger dissemination only in Bulgaria and the United Kingdom.
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COVID-19 , SARS-CoV-2 , Bulgaria/epidemiología , COVID-19/epidemiología , Genómica , Humanos , Mutación , Pandemias , SARS-CoV-2/genética , Glicoproteína de la Espiga del Coronavirus/genéticaRESUMEN
We illustrate the potential for specialist laboratory networks to be used as preparedness and response tool through rapid collection and sharing of data. Here, the Emerging Viral Diseases-Expert Laboratory Network (EVD-LabNet) and a laboratory assessment of chikungunya virus (CHIKV) in returning European travellers related to an ongoing outbreak in Thailand was used for this purpose. EVD-LabNet rapidly collected data on laboratory requests, diagnosed CHIKV imported cases and sequences generated, and shared among its members and with the European Centre for Disease Prevention and Control. Data across the network showed an increase in CHIKV imported cases during 1 October 2018-30 April 2019 vs the same period in 2018 (172 vs 50), particularly an increase in cases known to be related to travel to Thailand (72 vs 1). Moreover, EVD-LabNet showed that strains were imported from Thailand that cluster with strains of the ECSA-IOL E1 A226 variant emerging in Pakistan in 2016 and involved in the 2017 outbreaks in Italy. CHIKV diagnostic requests increased by 23.6% between the two periods. The impact of using EVD-LabNet or similar networks as preparedness and response tool could be improved by standardisation of the collection, quality and mining of data in routine laboratory management systems.
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Fiebre Chikungunya/epidemiología , Virus Chikungunya/aislamiento & purificación , Enfermedades Transmisibles Emergentes/prevención & control , Brotes de Enfermedades/prevención & control , Laboratorios/normas , Fiebre Chikungunya/diagnóstico , Notificación de Enfermedades , Humanos , Laboratorios/organización & administración , Filogenia , Tailandia/epidemiología , ViajeRESUMEN
BackgroundCrimean-Congo haemorrhagic fever virus (CCHFV) is considered an emerging infectious disease threat in the European Union. Since 2000, the incidence and geographic range of confirmed CCHF cases have markedly increased, following changes in the distribution of its main vector, Hyalomma ticks.AimsTo review scientific literature and collect experts' opinion to analyse relevant aspects of the laboratory management of human CCHF cases and any exposed contacts, as well as identify areas for advancement of international collaborative preparedness and laboratory response plans.MethodsWe conducted a literature review on CCHF molecular diagnostics through an online search. Further, we obtained expert opinions on the key laboratory aspects of CCHF diagnosis. Consulted experts were members of two European projects, EMERGE (Efficient response to highly dangerous and emerging pathogens at EU level) and EVD-LabNet (Emerging Viral Diseases-Expert Laboratory Network).ResultsConsensus was reached on relevant and controversial aspects of CCHF disease with implications for laboratory management of human CCHF cases, including biosafety, diagnostic algorithm and advice to improve lab capabilities. Knowledge on the diffusion of CCHF can be obtained by promoting syndromic approach to infectious diseases diagnosis and by including CCHFV infection in the diagnostic algorithm of severe fevers of unknown origin.ConclusionNo effective vaccine and/or therapeutics are available at present so outbreak response relies on rapid identification and appropriate infection control measures. Frontline hospitals and reference laboratories have a crucial role in the response to a CCHF outbreak, which should integrate laboratory, clinical and public health responses.
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Técnicas de Laboratorio Clínico/métodos , ADN Viral/genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/diagnóstico , Fiebre Hemorrágica de Crimea/transmisión , Ensayos de Aptitud de Laboratorios/normas , Animales , Enfermedades Transmisibles Emergentes/epidemiología , ADN Viral/análisis , Brotes de Enfermedades/prevención & control , Ensayo de Inmunoadsorción Enzimática , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica de Crimea/epidemiología , Fiebre Hemorrágica de Crimea/virología , Humanos , Inmunoglobulina G/sangre , Ixodidae , Laboratorios , Ensayos de Aptitud de Laboratorios/métodos , Análisis de Secuencia de ARN , Garrapatas/virologíaRESUMEN
IntroductionSequence-based typing of hepatitis A virus (HAV) is important for outbreak detection, investigation and surveillance. In 2013, sequencing was central to resolving a large European Union (EU)-wide outbreak related to frozen berries. However, as the sequenced HAV genome regions were only partly comparable between countries, results were not always conclusive.AimThe objective was to gather information on HAV surveillance and sequencing in EU/European Economic Area (EEA) countries to find ways to harmonise their procedures, for improvement of cross-border outbreak responses.MethodsIn 2014, the European Centre for Disease Prevention and Control (ECDC) conducted a survey on HAV surveillance practices in EU/EEA countries. The survey enquired whether a referral system for confirming primary diagnostics of hepatitis A existed as well as a central collection/storage of hepatitis A cases' samples for typing. Questions on HAV sequencing procedures were also asked. Based on the results, an expert consultation proposed harmonised procedures for cross-border outbreak response, in particular regarding sequencing. In 2016, a follow-up survey assessed uptake of suggested methods.ResultsOf 31 EU/EEA countries, 23 (2014) and 27 (2016) participated. Numbers of countries with central collection and storage of HAV positive samples and of those performing sequencing increased from 12 to 15 and 12 to 14 respectively in 2016, with all countries typing an overlapping fragment of 218 nt. However, variation existed in the sequenced genomic regions and their lengths.ConclusionsWhile HAV sequences in EU/EEA countries are comparable for surveillance, collaboration in sharing and comparing these can be further strengthened.
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Brotes de Enfermedades/prevención & control , Virus de la Hepatitis A/aislamiento & purificación , Hepatitis A/diagnóstico , Tipificación Molecular/métodos , Vigilancia de la Población/métodos , Secuenciación Completa del Genoma/métodos , Europa (Continente)/epidemiología , Unión Europea , Hepatitis A/epidemiología , Virus de la Hepatitis A/genética , Humanos , ARN Viral/análisis , Análisis de Secuencia de ADNRESUMEN
A case of a 66-year-old man with West Nile neuroinvassive disease manifested with fever, weakness, fatigue, consciousness disorders and underlying diabetes mellitus type 2 and cardiovascular diseases is presented. Laboratory data showed elevated erythrocyte sedimentation rate and fibrinogen. Serological tests revealed West Nile virus specific antibodies of class IgM and IgG in serum. West Nile virus RNA was detected in urine sample. Supportive therapy was applied.
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Monitoreo Epidemiológico , Enfermedades del Sistema Nervioso/epidemiología , Enfermedades del Sistema Nervioso/fisiopatología , Enfermedades del Sistema Nervioso/terapia , Fiebre del Nilo Occidental/fisiopatología , Fiebre del Nilo Occidental/terapia , Anciano , Bulgaria/epidemiología , Humanos , Masculino , Resultado del Tratamiento , Fiebre del Nilo Occidental/epidemiologíaRESUMEN
Crimean-Congo haemorrhagic fever (CCHF) is a severe tick-borne zoonotic disease in humans caused by CCHF virus. It has been observed in Bulgaria since 1952 and over the years more than 1600 cases have been reported in the country. Close contact with viraemic livestock was shown as one of the main causes of the infection. Detection of CCHF virus specific antibodies in livestock can be used as an indicator for virus circulation and risk assessment. CCHF seroprevalence was investigated in 843 cattle, 88 goats and 130 sheep, originating from all 28 districts of Bulgaria. CCHF virus-specific IgG antibodies were observed in 165 cattle (19.6, 95% CI 17.0-22.4%), in 20 goats (22.7, 95% CI 15.2-32.5%) and in 10 sheep (7.7, 95% CI 4.2-13.6%). The highest seroprevalence was detected in the district of Kardzhali, South Bulgaria (86.7, 95% CI 73.8-93.7%), a well-known focus of CCHF in humans. The other two districts with human CCHF cases, Blagoevgrad and Burgas, located in southwest and southeast Bulgaria, showed overall seroprevalences in livestock of 41.9% (95% CI 28.4-56.7%) and 31.3% (95% CI 22.2-42.1%), respectively. Seroprevalences in districts with no history of human CCHF cases varied between 55% (95% CI 39.8-69.3%) and 22.5% (95% CI 12.3-37.5%). These results suggest frequent CCHF virus infections even in regions without human CCHF cases and an enhanced risk of infection for humans in close contact with the infected livestock.
Asunto(s)
Enfermedades de los Bovinos/epidemiología , Enfermedades de las Cabras/epidemiología , Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/veterinaria , Enfermedades de las Ovejas/epidemiología , Animales , Bulgaria/epidemiología , Bovinos , Enfermedades de los Bovinos/virología , Enfermedades de las Cabras/virología , Cabras , Fiebre Hemorrágica de Crimea/epidemiología , Fiebre Hemorrágica de Crimea/virología , Prevalencia , Estudios Seroepidemiológicos , Ovinos , Enfermedades de las Ovejas/virología , Enfermedades por Picaduras de Garrapatas/epidemiología , Enfermedades por Picaduras de Garrapatas/virología , Zoonosis/epidemiología , Zoonosis/virologíaRESUMEN
To assess local circulation and risk for human infections with West Nile virus (WNV) and Tick-borne encephalitis virus (TBEV) in Bulgaria, a nationwide seroprevalence study was conducted. In total, 1451 residents of all 28 districts in Bulgaria were tested for WNV-specific and TBEV-specific IgG antibodies. The survey found overall seroprevalence of 1.5% and 0.6%, respectively. The highest WNV seroprevalence was found in Sofia Province and districts near the river Danube. TBEV circulation was detected among residents of six districts. The results showed that the two virus infections seem to be more wide-spread in the country as has been described.
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Anticuerpos Antivirales/sangre , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Encefalitis Transmitida por Garrapatas/epidemiología , Estudios Seroepidemiológicos , Fiebre del Nilo Occidental/epidemiología , Adulto , Anciano , Bulgaria/epidemiología , Encefalitis Transmitida por Garrapatas/virología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Grupos de Población , Factores de Riesgo , Fiebre del Nilo Occidental/inmunología , Fiebre del Nilo Occidental/virología , Virus del Nilo Occidental/aislamiento & purificaciónRESUMEN
Crimean-Congo hemorrhagic fever (CCHF) and hantavirus infections are the two viral hemorrhagic fevers spread in Europe. To test actual circulation of CCHF virus (CCHFV) and hantaviruses in Bulgaria, we conducted country-wide seroepidemiological studies. Serum samples were collected prospectively from 1500 residents of all 28 districts in Bulgaria. CCHFV seroprevalence of 3.7% was revealed. Anamnesis for tick bites, contact with livestock, age over 40 years and residency in Haskovo district were found as risk factors. The highest CCHFV seroprevalence was observed in the known endemic districts in southeastern Bulgaria: Haskovo (28%) and Yambol (12%). Reactive samples were found in residents of 20 of the 28 districts in Bulgaria. In comparison with the previous studies, the data presented indicate that CCHFV increased substantially its circulation in the endemic regions and was introduced in many new areas. Hantavirus seroprevalence was based on results of the immunoblot and estimated as 3.1%. Surprisingly, contrary to all available data, Puumala virus seroprevalence rate was 2.3% versus 0.8% of Dobrava-Belgrade virus. Evidence for hantavirus IgG seropositivity was found in residents of 23 of the 28 districts in the country. The first hantavirus seroprevalence study in Bulgaria showed that Puumala virus is probably more wide-spread in the country than Dobrava-Belgrade virus.
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Anticuerpos Antivirales/sangre , Infecciones por Hantavirus/epidemiología , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica de Crimea/epidemiología , Orthohantavirus/inmunología , Estudios Seroepidemiológicos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bulgaria/epidemiología , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Orthohantavirus/aislamiento & purificación , Infecciones por Hantavirus/inmunología , Infecciones por Hantavirus/virología , Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/inmunología , Fiebre Hemorrágica de Crimea/virología , Humanos , Inmunoglobulina G/sangre , Lactante , Masculino , Persona de Mediana Edad , Mordeduras de Garrapatas , Adulto JovenRESUMEN
In countries from which Crimean-Congo haemorrhagic fever (CCHF) is absent, the causative virus, CCHF virus (CCHFV), is classified as a hazard group 4 agent and handled in containment level (CL)-4. In contrast, most endemic countries out of necessity have had to perform diagnostic tests under biosafety level (BSL)-2 or -3 conditions. In particular, Turkey and several of the Balkan countries have safely processed more than 100 000 samples over many years in BSL-2 laboratories. It is therefore advocated that biosafety requirements for CCHF diagnostic procedures should be revised, to allow the tests required to be performed under enhanced BSL-2 conditions with appropriate biosafety laboratory equipment and personal protective equipment used according to standardized protocols in the countries affected. Downgrading of CCHFV research work from CL-4, BSL-4 to CL-3, BSL-3 should also be considered.
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Contención de Riesgos Biológicos/normas , Virus de la Fiebre Hemorrágica de Crimea-Congo/fisiología , Fiebre Hemorrágica de Crimea/prevención & control , Exposición Profesional/prevención & control , Animales , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica de Crimea/virología , Humanos , Exposición Profesional/normasRESUMEN
Crimean-Congo hemorrhagic fever (CCHF) is endemic in Bulgaria. During 2013-2014, 11 confirmed CCHF cases have been reported in the country (seven in 2013 and four in 2014). The present study provides the CCHF molecular epidemiology in Bulgaria based on all currently available S, M, and L RNA segment nucleotide sequences spanning the years 1978-2014. A relatively low genetic difference (0-6%, the maximum seen in the M RNA segment) was seen among the CCHFV sequences suggesting that a slow evolving CCHFV strain belonging to "Europe 1" clade is present in Bulgaria. Although the virus emerged in new foci during the recent years, it is more active in the established endemic foci which seem to offer the most suitable ecosystem and environment. Understanding the CCHF epidemiology and virus evolution is the basis for public health programs and vaccine design.
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Enfermedades Endémicas , Variación Genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/clasificación , Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/epidemiología , Fiebre Hemorrágica de Crimea/virología , Adolescente , Adulto , Anciano , Bulgaria/epidemiología , Niño , Femenino , Genoma Viral , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Nairovirus , Análisis de Secuencia de ADN , Adulto JovenRESUMEN
Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.
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Antígenos Bacterianos/inmunología , Leptospirosis/diagnóstico , Polisacáridos/inmunología , Juego de Reactivos para Diagnóstico/normas , Argentina , Bulgaria , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Leptospira/aislamiento & purificación , Leptospira/metabolismo , Leptospira interrogans/aislamiento & purificación , Leptospira interrogans/metabolismo , Leptospirosis/microbiología , Masculino , República de Corea , Sensibilidad y EspecificidadRESUMEN
Crimean-Congo hemorrhagic fever (CCHF) is a human disease with high fatality rate. Although its pathogenesis is not elucidated yet, it is considered that cytokines play a significant role in the progression and outcome of the disease. Serum CXCL10 levels were estimated in 35 patients with acute CCHF and were correlated with the viral load, and various demographic and clinical parameters. The mean CXCL10 concentration in the patients' group was higher compared to the respective value in the control group (4421.74 pg/ml vs. 28.47 pg/ml, P < 0.05). A strong positive correlation between CXCL10 and viral load was seen (rs = 0.57, P < 0.001), while the outcome of the disease was related with the viral load (rs = 0.47, P = 0.004) and the presence of hemorrhagic manifestations (P < 0.001). The study provides an insight into the strong correlation between CXCL10 and viral load in acute CCHF cases suggesting that it plays an important role in CCHF pathogenesis.
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Quimiocina CXCL10/sangre , Virus de la Fiebre Hemorrágica de Crimea-Congo/fisiología , Fiebre Hemorrágica de Crimea/inmunología , Fiebre Hemorrágica de Crimea/virología , Carga Viral , Adolescente , Adulto , Anciano , Anticuerpos Antivirales/sangre , Quimiocina CXCL10/inmunología , Niño , Preescolar , Femenino , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica de Crimea/mortalidad , Fiebre Hemorrágica de Crimea/patología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto JovenRESUMEN
Several Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in Europe: Dobrava-Belgrade virus (DOBV), Puumala, Saaremaa, Sochi, and Seoul virus. Although HFRS is endemic in Bulgaria, genome sequences of hantaviruses have never been detected in wild rodents. To identify rodent reservoirs, a total of 691 rodents from three endemic regions were trapped in 2011-2012 and screened by TaqMan RT-PCR for detection of hantaviral genomic RNA. Partial small (S) and/or large (L)-segment sequences were recovered from six Apodemus mice: five of the species A. flavicollis and one A. agrarius. Phylogenetic analysis revealed that all recovered sequences belonged to DOBV. On the phylogenetic trees, the novel Bulgarian hantavirus sequences clustered together with sequences of established previously DOBV variants recovered from Bulgarian HFRS patients and also with variants found in wild rodents trapped in Slovenia, Greece, and Slovakia. One of the novel Bulgarian DOBV S-sequences from A. agrarius was related closely to DOBV sequences recovered from A. flavicollis, suggesting a spillover of DOBV from its natural host to A. agrarius mice. The results of this study confirmed the circulation of DOBV in wild rodents in Bulgaria. The complexity of the epidemiological situation in the Balkans requires further studies of hantaviruses in rodent hosts and human HFRS cases.
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Reservorios de Enfermedades , Murinae/virología , Orthohantavirus/aislamiento & purificación , ARN Viral/aislamiento & purificación , Animales , Bulgaria , Análisis por Conglomerados , Orthohantavirus/clasificación , Orthohantavirus/genética , Ratones , Filogenia , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
Crimean-Congo haemorrhagic fever (CCHF) is a tick-borne zoonotic disease. Over the past decade, CCHF cases in humans have emerged in Turkey and reemerged in the Balkan countries, Ukraine and Tajikistan. Occupational contact with infected livestock has been recognized as a common cause of the disease. A cross-sectional seroprevalence study in livestock was conducted in farming communities of an endemic area in Bulgaria, southeastern Europe. Overall, 72% of the tested animals were positive for IgG antibodies to CCHF virus. By the time the animals were one-year old almost 50% had serologic evidence of CCHF infection, and by two years already 80% of them had been infected. The data obtained in this study reflect current situation of CCHF virus infection among livestock in Bulgaria. The results showed active CCHF virus circulation that poses risk for humans to be infected during contacts with animals and requires public health awareness.
RESUMEN
Introduction. After two seasons of absence and low circulation, influenza activity increased significantly in the winter of 2022-2023. This study aims to characterize virological and epidemiological aspects of influenza infection in Bulgaria during the 2022-2023 season and perform a phylogenetic/molecular analysis of the hemagglutinin (HA) and neuraminidase (NA) sequences of representative influenza strains.Hypothesis/Gap Statement. Influenza A and B viruses generate new genetic groups/clades each season, replacing previously circulating variants. This results in increased antigenic distances from current vaccine strains. Strengthening existing influenza surveillance is essential to meet the challenges posed by the co-circulation of influenza and SARS-CoV-2.Methodology. We tested 2713 clinical samples from patients with acute respiratory illnesses using a multiplex real-time RT-PCR kit (FluSC2) to detect influenza A/B and Severe acute respiratory syndrome coronavirus-2(SARS-CoV-2) simultaneously. Representative Bulgarian influenza strains were sequenced at the WHO Collaborating Centres in London, UK, and Atlanta, USA.Results. Influenza virus was detected in 694 (25.6â%) patients. Of these, 364 (52.4â%), 213 (30.7â%) and 117 (16.9â%) were positive for influenza A(H1N1)pdm09, A(H3N2) and B/Victoria lineage virus, respectively. HA genes of the 47 influenza A(H1N1)pdm09 viruses fell into clades 5a.2. and 5a.2a.1 within the 6B.5A.1A.5a.2 group. Twenty-seven A(H3N2) viruses belonging to subclades 2b, 2a.1, 2a.1b and 2a.3a.1 within the 3C.2a1b.2a.2 group were analysed. All 23 sequenced B/Victoria lineage viruses were classified into the V1A.3a.2 group. We identified amino acid substitutions in HA and NA compared with the vaccine strains, including several substitutions in the HA antigenic sites.Conclusion. The study's findings showed genetic diversity among the influenza A viruses and, to a lesser extent, among B viruses, circulating in the first season after the lifting of anti-COVID-19 measures.
Asunto(s)
Variación Genética , Virus de la Influenza B , Gripe Humana , Neuraminidasa , Filogenia , SARS-CoV-2 , Humanos , Gripe Humana/virología , Gripe Humana/epidemiología , Virus de la Influenza B/genética , Virus de la Influenza B/clasificación , Virus de la Influenza B/aislamiento & purificación , SARS-CoV-2/genética , SARS-CoV-2/clasificación , Neuraminidasa/genética , Adulto , Masculino , Persona de Mediana Edad , Femenino , Bulgaria/epidemiología , Adulto Joven , Anciano , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Preescolar , Niño , Adolescente , COVID-19/epidemiología , COVID-19/virología , Lactante , Estaciones del Año , Virus de la Influenza A/genética , Virus de la Influenza A/clasificación , Virus de la Influenza A/aislamiento & purificación , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/clasificación , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Subtipo H3N2 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/clasificación , Subtipo H3N2 del Virus de la Influenza A/aislamiento & purificaciónRESUMEN
Background: Respiratory syncytial virus (RSV) is a common cause of upper and lower respiratory tract infections. This study aimed to explore the prevalence of respiratory syncytial virus (RSV) and other respiratory viruses in Bulgaria, characterize the genetic diversity of RSV strains, and perform amino acid sequence analyses of RSV surface and internal proteins. Methods: Clinical and epidemiological data and nasopharyngeal swabs were prospectively collected from patients with acute respiratory infections between October 2020 and May 2023. Real-time PCR for 13 respiratory viruses, whole-genome sequencing, phylogenetic, and amino acid analyses were performed. Results: This study included three epidemic seasons (2020-2021, 2021-2022, and 2022-2023) from week 40 of the previous year to week 20 of the following year. Of the 3,047 patients examined, 1,813 (59.5%) tested positive for at least one viral respiratory pathogen. RSV was the second most detected virus (10.9%) after SARS-CoV-2 (22%). Coinfections between RSV and other respiratory viruses were detected in 68 cases, including 14 with SARS-CoV-2. After two seasons of low circulation, RSV activity increased significantly during the 2022-2023 season. The detection rates of RSV were 3.2, 6.6, and 13.7% in the first, second, and third seasons, respectively. RSV was the most common virus found in children under 5 years old with bronchiolitis (40%) and pneumonia (24.5%). RSV-B drove the 2022-2023 epidemic. Phylogenetic analysis indicated that the sequenced RSV-B strains belonged to the GB5.0.5a and GB5.0.6a genotypes. Amino acid substitutions in the surface and internal proteins, including the F protein antigenic sites were identified compared to the BA prototype strain. Conclusion: This study revealed a strong resurgence of RSV in the autumn of 2022 after the lifting of anti-COVID-19 measures, the leading role of RSV as a causative agent of serious respiratory illnesses in early childhood, and relatively low genetic diversity in circulating RSV strains.
RESUMEN
This study aimed to determine the incidence and etiological, seasonal, and genetic characteristics of respiratory viral coinfections involving severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Between October 2020 and January 2024, nasopharyngeal samples were collected from 2277 SARS-CoV-2-positive patients. Two multiplex approaches were used to detect and sequence SARS-CoV-2, influenza A/B viruses, and other seasonal respiratory viruses: multiplex real-time polymerase chain reaction (PCR) and multiplex next-generation sequencing. Coinfections of SARS-CoV-2 with other respiratory viruses were detected in 164 (7.2%) patients. The most common co-infecting virus was respiratory syncytial virus (RSV) (38 cases, 1.7%), followed by bocavirus (BoV) (1.2%) and rhinovirus (RV) (1.1%). Patients ≤ 16 years of age had the highest rate (15%) of mixed infections. Whole-genome sequencing produced 19 complete genomes of seasonal respiratory viral co-pathogens, which were subjected to phylogenetic and amino acid analyses. The detected influenza viruses were classified into the genetic groups 6B.1A.5a.2a and 6B.1A.5a.2a.1 for A(H1N1)pdm09, 3C.2a1b.2a.2a.1 and 3C.2a.2b for A(H3N2), and V1A.3a.2 for the B/Victoria lineage. The RSV-B sequences belonged to the genetic group GB5.0.5a, with HAdV-C belonging to type 1, BoV to genotype VP1, and PIV3 to lineage 1a(i). Multiple amino acid substitutions were identified, including at the antibody-binding sites. This study provides insights into respiratory viral coinfections involving SARS-CoV-2 and reinforces the importance of genetic characterization of co-pathogens in the development of therapeutic and preventive strategies.