Hantavirus infection has been a notifiable disease since 2008in Hong Kong. A total of 44 cases were reported from 1995 to 2013. Rodents are the major hosts of pathogenic hantaviruses. However, the epidemiology data about hantavirus infection in rodents is not known. The present study aims to investigate the prevalence of hantavirus infection in rodents and strains of hantaviruses prevailing in Hong Kong. A total of 502 street rodents were collected from various districts between October 2008 and July 2013 and the majority was Rattus norvegicus. Spleen and kidney tissues were extracted to perform RT-PCR. Among 1004 tissue samples, hantaviruses were detected in 15 samples (1.49%) from 11 rodents (2.19%). The hantavirus positive rodents were widely distributed in Hong Kong in which most cases were found from Yuen Long and Wong Tai Sin. Hantavirus infection cases were mainly reported in early-spring and summer. Phylogenetic analysis of the partial nucleocapsid protein gene of the positive rodent sequences revealed that all of them belonged to Seoul virus (SEOV). Three strains of SEOV which were genetically close to strains in China, Indonesia and Vietnam, were identified. / published_or_final_version / Microbiology / Master / Master of Medical Sciences

Thesis (M.S.)--University of Nevada, Reno, 2008. / "May, 2008." Includes bibliographical references. Online version available on the World Wide Web.

Thesis (M. Sc.)--Carleton University, 1997. / Includes bibliographical references. Also available in electronic format on the Internet.

Thesis (Ph. D.)--University of Hawaii at Manoa, 1990. / Includes bibliographical references (leaves 92-108) / Microfiche. / ix, 108 leaves, bound ill. 29 cm

As hantaviroses são zoonoses de roedores silvestres que causam doenças humanas graves: febre hemorrágica com síndrome renal na Ásia e Europa, com letalidade de 10 a 12% e no continente americano causam a Síndrome Pulmonar e Cardiovascular por Hantavirus (SPCVH), letalidade de 59%. Os Hantavirus (família Bunyaviridae) são vírus envelopados que medem aproximadamente 120 nm, possuem RNA de fita simples e polaridade negativa, dividida em 3 segmentos (L, M e S) que se replicam no citoplasma. A infecção humana relaciona-se à inalação de aerossóis de excretas de roedores infectados com Hantavirus, embora existam relatos de transmissão interpessoal na América do Sul. No Estado de São Paulo, os roedores infectados mais encontrados foram o “rato do rabo peludo" (Bolomys lasiurus), o “rato da mata" (Akodon cursor) e o “ratinho do arroz" (Oligoryzomys negrips). Desde os 3 primeiros casos de hantavirose descritos no Brasil (1993) com indivíduos moradores da área rural de Juquitiba, SP, evidenciando o primeiro surto conhecido de SPCVH, mais de uma centena de casos da SPCVH foram notificados, causando 60 óbitos (letalidade de 46%). Na região de Ribeirão Preto, ocorreram quatorze casos de SPCVH de 1998 a 2001, com letalidade de 64,2%, o que motivou este trabalho. Um primeiro objetivo deste trabalho foi estudar aspectos clínicos e epidemiológicos dos casos de hantavirose ocorridos na região de Ribeirão Preto, de 1998 a 2001, para conhecer o comportamento das hantaviroses nesta região. Analisando e acompanhando prontuários de 14 pacientes com SPCVH, observou-se febre (100% dos casos), estertores pulmonares, dispnéia e tosse (64,2%), taquicardia acompanhada de hipotensão (64,2%), cefaléia e sintomas digestivos (57%), adinamia e indisposição geral (50%), fenômenos hemorrágicos (28,5%), mialgia (21,4%) e convulsão (7,5%). No perfil laboratorial, a plaquetopenia <130000/mm3 (100%), hematócrito >55%, (78,6%); leucocitose >15000/mm3 (64,2%), neutrofilia >7000/ mm3 e bastonetes >600 cels/mm3 (64,2%), creatinina sérica >1,5mg/100ml (63,6%) e PO2 <70 mmHg (54,5%). O diagnóstico laboratorial das hantaviroses foi feito por ELISA para IgM e IgG anti Sin Nombre após a alta. Houve diferença significativa entre casos fatais e sobreviventes, quanto à época de suspeita diagnóstica de hantavirose (p=0,0152) e quanto ao uso de hidratação parenteral (p=0,0152): nos sobreviventes, a suspeita diagnóstica de SPCVH foi feita entre o 1º e o 2º dia de internação e, nos casos fatais, a suspeita foi feita apenas na necropsia. Existe uma correlação significativa entre o volume de infusão endovenosa de líquidos &#61619; 2500 ml, nas primeiras 24h de tratamento, e a evolução dos casos para o óbito. A oxigenação precoce, ventilação mecânica, assim como o uso de aminas vasoativas e o de corticosteróides não tiveram associação com sobrevida e a presença de choque não teve associação com óbito. O segundo objetivo foi estudar a prevalência de anticorpos para Hantavirus na população de Jardinópolis, na região de Ribeirão Preto, SP. Em estudo prévio entre cidades da região de Ribeirão Preto, obteve-se maior índice de prevalência sorológica (4,5%) para Hantavirus naquele município. Através de coleta aleatória em parte da área rural e em toda a área urbana, obtivemos 818 amostras de sangue de moradores entre 15 e 70 anos, por digitopuntura em papel de filtro, dos quais coletamos dados relevantes como idade, sexo, cor, procedência, atividade profissional, endereço, telefone; moléstia atual, medicamentos em uso, pneumonia grave pregressa, tipo de moradia, número de moradores na casa, esgoto, coleta de lixo, celeiro, contato com roedores. Essas amostras foram processadas nas diluições 1/50, 1/100, 1/400 e 1/1600 e submetidas ao ELISA indireto para detecção de IgG para Hantavirus Andes. Os resultados foram analisados pelo teste de Fisher e teste do Qui-quadrado, utilizando o programa InStat 3.0 (GraphPad Software Inc, USA) e para &#945; de 5%, com intervalo de confiança de 95%. A positividade geral foi de 14,3% com ELISA à diluição 1/50, na zona urbana foi de 15,3% e na zona rural de 6,5%, demonstrando alta representatividade para a população do município de Jardinópolis. Entre moradores de zona urbana e rural, a positividade para Hantavirus foi significativamente maior para os moradores urbanos (p= 0,0183). E entre as diferentes faixas etárias com a das restantes, observou-se diminuição significativa para a faixa etária de 21 a 30 anos (p= 0,0117) e aumentada para a de 31 a 40 anos (p= 0,0036). Não houve diferença significativa de positividade para Hantavirus observada nos homens e nas mulheres (p= 0,0892), nem para indivíduos com atividade profissional urbana e rural (p=0,8799), nem com os indivíduos que referiram pneumonia grave (p= 0,6096) e da mesma forma nos indivíduos que declararam contato com roedores (p= 0,4842). Assim como não houve associação entre títulos de 100 e 400 para as mesmas características analisadas. / The hantaviruses are zoonoses of wild rodents that cause serious human diseases: hemorrhagic fever with renal syndrome in Asia and Europe (10 to 12% lethality rates) and Pulmonary and Cardiovascular Syndrome in the American continent (HPCVS - 59% lethality rate). Hantaviruses (family Bunyaviridae) are enclosing viruses that measure 120 nm approximately, they possess RNA of simple ribbon and negative sense, divided in 3 segments (L, M and S) that are replication in the cytoplasm. The human infection links to the inhalation of aerosols of Hantavirus infected rodents excretes, although reports of person-to-person transmission exist in South America. In the State of São Paulo the infected rodents found were the Bolomys lasiurus, the Akodon cursor and the Oligoryzomys negrips. From the first 3 cases of hantaviruses described in Brazil (1993) with individuals inhabitants of the rural area of Juquitiba, SP, evidencing the first well-known supplies of HPCVS, more than a hundred cases have been notified in Brazil, causing 60 deaths (of 46% lethality rate). There have been fourteen cases of HPCVS since1998 to 2001 with of 64,2% letality rate, in the Ribeirão Preto area what motivated this work. One of the objectives was clinical and epidemic aspects studies of the cases of hantaviruses that happened in the Area of Ribeirão Preto, from 1998 to 2001, to know the behavior of the hantavirusess in this area. Analyzing and accompanying patient medical records of 14 patients with HPCVS, fever was observed (100% of the cases), disnea and coughs (64,2%), accompanied of arterial hipotension, tachicardia (64,2%), migraine and digestive symptoms (57%); adynamic and general weakness (50%), hemorrhagic phenomenons (28,5%), mialgia (21,4%) and convulsion (7,5%). In the laboratorial profile the platelets count <130000/mm3 (100%), hematocrit> 55%, (78,6%); leucocytosis > 15000/mm3 (64,2%), neutrofilia >7000 / mm3 and small sticks> 600 cels / mm3 (64,2%), creatinine serical level > 1,5mg/100ml (63,6%) and PO2 <70 mmHg (54,5%). The diagnosis laboratorial of the hantaviruses was made by ELISA for IgM and IgG anti Sin Nombre after the discharge. There was significant difference among fatal and surviving cases, as the hantaviruses diagnosis suspicious (p=0,0152) and parenteral hydratation (p=0,0152): in the survivors, the HPCVS diagnosis suspicious was made between the 1st and the 2nd day of internment and the fatal cases, the suspicion was just made to the necropsies. Among infusion of liquid EV ³ over 2500 ml in the first 24 hours of treatment and evolution for the death. The precocious oxygen support, mechanics ventilation, as well as the use of vasoatives amines and the one of corticosteróides, did not have association with survival and the shock presence did not have association with death. The second objective was to antibodies for Hantavirus prevalence studies in the Area of Ribeirão Preto, in the population of Jardinópolis, SP. In the previous study among cities in that area it was obtained larger index of serologic prevalence (4,5%) for Hantavirus, it was in the Jardinópolis county Through ramdomised collection partly of the rural area and whole urban area, we obtained 818 samples of blood, inhabitants between 15 and 70 years, for digital puncture in filter paper and important data as age, gender, color, origin, professional activity, address, telephone; current disease, medications in use, pneumonia serious, dwelling type, number of inhabitants in the house, sewer, collects of garbage, barn, contact with rodents. These samples were processed, in the dilutions 1/50, 1/100, 1/400 and 1/1600. and submitted to indirect ELISA for detection of IgG for Hantavirus Los Andes. The results were analyzed by Fisher and Qui-square tests, using the InStat 3.0 program (GraphPad.Software Inc, USA) and for the one of 5%, with interval of trust of 95%. The general positivity rated 14,3% with ELISA (at title 50), urban zone (15,3%) and in rural zone 6,5% and they demonstrated high rates for the population of the Jardinópolis county. Comparing the positivity rates for Hantavirus between urban and rural zone inhabitants was significantly larger for the urban inhabitants (p = 0,0183). Significant increase was observed for the age group of 21 to 30 years (p = 0,0117) and for the one of 31 to 40 years (p = 0,0036) among different age groups. There was not significant difference of positivist for Hantavirus between men and women (p = 0,0892); nor for individuals with urban and rural professional activity (p=0,8799), neither for the individuals that referred serious pneumonia (p = 0,6096) as well as in the individuals that declared contact with rodents (p = 0,4842). There was not association among titles of 100 and 400 for Hantavirus with gender, local of habitation, contact with rodents and pneumonia serious.

"Sin Nombre virus is a hantavirus first recognized in New Mexico in 1993. This virus is responsible for causing Hantavirus Pulmonary Syndrome, an acute, life threatening illness characterized by pulmonary edema, capillary leaking, and extreme respiratory distress. CD8+ cytotoxic T-cell lines specific for Sin Nombre virus were isolated from the peripheral blood mononuclear cells (PBMC) of a donor (NM3) who was naturally infected with the Sin Nombre virus, and has survived hantavirus pulmonary syndrome (HPS). Cytotoxic T lymphocyte (CTL) assays showed that one of these cell lines, 10K, specifically recognizes a nine amino acid epitope, TAHGVGIIP (amino acids 664-672 of the precursor GPC protein), which is located in the G2 protein after cleavage. Another cell line, 10c27, specifically recognized an eight amino acid epitope, AHGVGIIP (amino acids 665-672 of the precursor GPC protein), located in the G2 protein after cleavage. Using polymerase chain reaction (PCR) and CTL assays, the recognition of these epitopes was shown to be restricted by the B35.01 Class 1 human leukocyte associated antigen (HLA) allele. This information will be useful in creating a vaccine for use in immunizing people against the Sin Nombre hantavirus, as well as elucidating the pathogenesis of this disease. "

As hantaviroses são zoonoses de roedores silvestres que causam doenças humanas graves: febre hemorrágica com síndrome renal na Ásia e Europa, com letalidade de 10 a 12% e no continente americano causam a Síndrome Pulmonar e Cardiovascular por Hantavirus (SPCVH), letalidade de 59%. Os Hantavirus (família Bunyaviridae) são vírus envelopados que medem aproximadamente 120 nm, possuem RNA de fita simples e polaridade negativa, dividida em 3 segmentos (L, M e S) que se replicam no citoplasma. A infecção humana relaciona-se à inalação de aerossóis de excretas de roedores infectados com Hantavirus, embora existam relatos de transmissão interpessoal na América do Sul. No Estado de São Paulo, os roedores infectados mais encontrados foram o “rato do rabo peludo” (Bolomys lasiurus), o “rato da mata” (Akodon cursor) e o “ratinho do arroz” (Oligoryzomys negrips). Desde os 3 primeiros casos de hantavirose descritos no Brasil (1993) com indivíduos moradores da área rural de Juquitiba, SP, evidenciando o primeiro surto conhecido de SPCVH, mais de uma centena de casos da SPCVH foram notificados, causando 60 óbitos (letalidade de 46%). Na região de Ribeirão Preto, ocorreram quatorze casos de SPCVH de 1998 a 2001, com letalidade de 64,2%, o que motivou este trabalho. Um primeiro objetivo deste trabalho foi estudar aspectos clínicos e epidemiológicos dos casos de hantavirose ocorridos na região de Ribeirão Preto, de 1998 a 2001, para conhecer o comportamento das hantaviroses nesta região. Analisando e acompanhando prontuários de 14 pacientes com SPCVH, observou-se febre (100% dos casos), estertores pulmonares, dispnéia e tosse (64,2%), taquicardia acompanhada de hipotensão (64,2%), cefaléia e sintomas digestivos (57%), adinamia e indisposição geral (50%), fenômenos hemorrágicos (28,5%), mialgia (21,4%) e convulsão (7,5%). No perfil laboratorial, a plaquetopenia <130000/mm3 (100%), hematócrito >55%, (78,6%); leucocitose >15000/mm3 (64,2%), neutrofilia >7000/ mm3 e bastonetes >600 cels/mm3 (64,2%), creatinina sérica >1,5mg/100ml (63,6%) e PO2 <70 mmHg (54,5%). O diagnóstico laboratorial das hantaviroses foi feito por ELISA para IgM e IgG anti Sin Nombre após a alta. Houve diferença significativa entre casos fatais e sobreviventes, quanto à época de suspeita diagnóstica de hantavirose (p=0,0152) e quanto ao uso de hidratação parenteral (p=0,0152): nos sobreviventes, a suspeita diagnóstica de SPCVH foi feita entre o 1º e o 2º dia de internação e, nos casos fatais, a suspeita foi feita apenas na necropsia. Existe uma correlação significativa entre o volume de infusão endovenosa de líquidos &#61619; 2500 ml, nas primeiras 24h de tratamento, e a evolução dos casos para o óbito. A oxigenação precoce, ventilação mecânica, assim como o uso de aminas vasoativas e o de corticosteróides não tiveram associação com sobrevida e a presença de choque não teve associação com óbito. O segundo objetivo foi estudar a prevalência de anticorpos para Hantavirus na população de Jardinópolis, na região de Ribeirão Preto, SP. Em estudo prévio entre cidades da região de Ribeirão Preto, obteve-se maior índice de prevalência sorológica (4,5%) para Hantavirus naquele município. Através de coleta aleatória em parte da área rural e em toda a área urbana, obtivemos 818 amostras de sangue de moradores entre 15 e 70 anos, por digitopuntura em papel de filtro, dos quais coletamos dados relevantes como idade, sexo, cor, procedência, atividade profissional, endereço, telefone; moléstia atual, medicamentos em uso, pneumonia grave pregressa, tipo de moradia, número de moradores na casa, esgoto, coleta de lixo, celeiro, contato com roedores. Essas amostras foram processadas nas diluições 1/50, 1/100, 1/400 e 1/1600 e submetidas ao ELISA indireto para detecção de IgG para Hantavirus Andes. Os resultados foram analisados pelo teste de Fisher e teste do Qui-quadrado, utilizando o programa InStat 3.0 (GraphPad Software Inc, USA) e para &#945; de 5%, com intervalo de confiança de 95%. A positividade geral foi de 14,3% com ELISA à diluição 1/50, na zona urbana foi de 15,3% e na zona rural de 6,5%, demonstrando alta representatividade para a população do município de Jardinópolis. Entre moradores de zona urbana e rural, a positividade para Hantavirus foi significativamente maior para os moradores urbanos (p= 0,0183). E entre as diferentes faixas etárias com a das restantes, observou-se diminuição significativa para a faixa etária de 21 a 30 anos (p= 0,0117) e aumentada para a de 31 a 40 anos (p= 0,0036). Não houve diferença significativa de positividade para Hantavirus observada nos homens e nas mulheres (p= 0,0892), nem para indivíduos com atividade profissional urbana e rural (p=0,8799), nem com os indivíduos que referiram pneumonia grave (p= 0,6096) e da mesma forma nos indivíduos que declararam contato com roedores (p= 0,4842). Assim como não houve associação entre títulos de 100 e 400 para as mesmas características analisadas. / The hantaviruses are zoonoses of wild rodents that cause serious human diseases: hemorrhagic fever with renal syndrome in Asia and Europe (10 to 12% lethality rates) and Pulmonary and Cardiovascular Syndrome in the American continent (HPCVS - 59% lethality rate). Hantaviruses (family Bunyaviridae) are enclosing viruses that measure 120 nm approximately, they possess RNA of simple ribbon and negative sense, divided in 3 segments (L, M and S) that are replication in the cytoplasm. The human infection links to the inhalation of aerosols of Hantavirus infected rodents excretes, although reports of person-to-person transmission exist in South America. In the State of São Paulo the infected rodents found were the Bolomys lasiurus, the Akodon cursor and the Oligoryzomys negrips. From the first 3 cases of hantaviruses described in Brazil (1993) with individuals inhabitants of the rural area of Juquitiba, SP, evidencing the first well-known supplies of HPCVS, more than a hundred cases have been notified in Brazil, causing 60 deaths (of 46% lethality rate). There have been fourteen cases of HPCVS since1998 to 2001 with of 64,2% letality rate, in the Ribeirão Preto area what motivated this work. One of the objectives was clinical and epidemic aspects studies of the cases of hantaviruses that happened in the Area of Ribeirão Preto, from 1998 to 2001, to know the behavior of the hantavirusess in this area. Analyzing and accompanying patient medical records of 14 patients with HPCVS, fever was observed (100% of the cases), disnea and coughs (64,2%), accompanied of arterial hipotension, tachicardia (64,2%), migraine and digestive symptoms (57%); adynamic and general weakness (50%), hemorrhagic phenomenons (28,5%), mialgia (21,4%) and convulsion (7,5%). In the laboratorial profile the platelets count <130000/mm3 (100%), hematocrit> 55%, (78,6%); leucocytosis > 15000/mm3 (64,2%), neutrofilia >7000 / mm3 and small sticks> 600 cels / mm3 (64,2%), creatinine serical level > 1,5mg/100ml (63,6%) and PO2 <70 mmHg (54,5%). The diagnosis laboratorial of the hantaviruses was made by ELISA for IgM and IgG anti Sin Nombre after the discharge. There was significant difference among fatal and surviving cases, as the hantaviruses diagnosis suspicious (p=0,0152) and parenteral hydratation (p=0,0152): in the survivors, the HPCVS diagnosis suspicious was made between the 1st and the 2nd day of internment and the fatal cases, the suspicion was just made to the necropsies. Among infusion of liquid EV ³ over 2500 ml in the first 24 hours of treatment and evolution for the death. The precocious oxygen support, mechanics ventilation, as well as the use of vasoatives amines and the one of corticosteróides, did not have association with survival and the shock presence did not have association with death. The second objective was to antibodies for Hantavirus prevalence studies in the Area of Ribeirão Preto, in the population of Jardinópolis, SP. In the previous study among cities in that area it was obtained larger index of serologic prevalence (4,5%) for Hantavirus, it was in the Jardinópolis county Through ramdomised collection partly of the rural area and whole urban area, we obtained 818 samples of blood, inhabitants between 15 and 70 years, for digital puncture in filter paper and important data as age, gender, color, origin, professional activity, address, telephone; current disease, medications in use, pneumonia serious, dwelling type, number of inhabitants in the house, sewer, collects of garbage, barn, contact with rodents. These samples were processed, in the dilutions 1/50, 1/100, 1/400 and 1/1600. and submitted to indirect ELISA for detection of IgG for Hantavirus Los Andes. The results were analyzed by Fisher and Qui-square tests, using the InStat 3.0 program (GraphPad.Software Inc, USA) and for the one of 5%, with interval of trust of 95%. The general positivity rated 14,3% with ELISA (at title 50), urban zone (15,3%) and in rural zone 6,5% and they demonstrated high rates for the population of the Jardinópolis county. Comparing the positivity rates for Hantavirus between urban and rural zone inhabitants was significantly larger for the urban inhabitants (p = 0,0183). Significant increase was observed for the age group of 21 to 30 years (p = 0,0117) and for the one of 31 to 40 years (p = 0,0036) among different age groups. There was not significant difference of positivist for Hantavirus between men and women (p = 0,0892); nor for individuals with urban and rural professional activity (p=0,8799), neither for the individuals that referred serious pneumonia (p = 0,6096) as well as in the individuals that declared contact with rodents (p = 0,4842). There was not association among titles of 100 and 400 for Hantavirus with gender, local of habitation, contact with rodents and pneumonia serious.

Hantaviren sind einzelsträngige RNA-Viren, die zur Familie der Bunyaviridae gehören. In den letzten Jahren haben sie immer mehr Aufmerksamkeit als “emerging viruses” auf sich gezogen, welche zunehmend Relevanz als humanpathogene Erreger gewinnen. Sie verursachen, abhängig vom Hantavirustyp, zwei verschiedene Krankheitsbilder: das hämorrhagische Fieber mit renalem Syndrom (HFRS) und das kardiopulmonale Syndrom (HCPS). Beide Syndrome sind mit Veränderungen in der vaskulären Permeabilität, akutem Abfall der Blutplättchen (Thrombozytopenie) und Koagulationsdefekten verbunden. Die zugrundeliegenden Mechanismen sind noch immer kaum verstanden. Hantaviren infizieren zwar Endothelzellen, bisher aber konnte kein zytopathischer Effekt nachgewiesen werden. Diese Doktorarbeit untersucht in vitro, ob und auf welche Weise Hantaviren mit Zellen des hämostatischen Systems interagieren. Zu diesen gehören insbesondere Megakaryozyten, die Thrombozyten generieren und die Thrombozyten selbst. Wir zeigen, dass das pathogene Hantaan Virus (HTNV), im Gegensatz zu dem weniger pathogenen Tula Virus (TULV) und Prospect Hill Virus (PHV), megakaryozytäre Zellen infiziert. Interessanterweise erhöht sich nach Induktion der Differenzierung in den infizierten megakaryozytären Zellen die Virusrepliaktion drastisch. Das Überleben der Zellen und der Verlauf der Differenzierung wird dadurch jedoch nicht beeinflusst. Ähnlich, wie bereits früher für Endothelzellen gezeigt, haben demnach pathogene Hantaviren keinen direkten zytopathischen Effekt auf Megakaryozyten. Weiterhin verdeutlicht die vorliegende Doktorarbeit, dass HTNV mit humanen Thrombozyten interagiert. Dies resultiert in einer Herunterregulation von essentiellen Oberflächenmarkern, die wichtig für die Aggregation und das Signalling sind. / Hantaviruses are single stranded (ss) RNA viruses belonging to the family Bunyaviridae. Over the past years they attracted more and more attention as “emerging viruses”, increasingly gaining relevance as human pathogens. Depending on the hantavirus type they cause haemorrhagic fever with renal syndrome (HFRS) or hantavirus cardiopulmonary syndrome (HCPS). Both syndromes are associated with changes in vascular permeability, acute thrombocytopenia, and defects in platelet function. The underlying mechanisms are still poorly understood. Although hantaviruses infect endothelial cells, no viral cytopathic effect after infection was observed. This thesis investigates in vitro whether and how hantaviruses target cells of the haemostatic system. In particular megakaryocytes (MKs), which generate platelets, and platelets themselves. We show that pathogenic Hantaan virus (HTNV), in contrast to less pathogenic Tula virus (TULV) and Prospect Hill virus (PHV), infects megakaryocytic cells. Intriguingly, after induction of differentiation megakaryocytic cells switch from low-level to high-level HTNV production without reduction in cell survival or alteration in differentiation. Thus, there is no direct viral pathogenic effect on megakaryocytic cells as previously observed for endothelial cells. Furthermore, this study demonstrates that HTNV interacts with human platelets, resulting in downregulation of essential adhesion markers, which are important for platelet aggregation and signalling.

Thesis (MScMedSc (Pathology. Medical Virology))--University of Stellenbosch, 2010. / Bibliography / ENGLISH ABSTRACT: Throughout history disease entities have been described which match the description of diseases now known to be caused by hantaviruses; however these viruses were first identified as the aetiologic agent in 1976, the first species named Hantaan virus after the river near which its natural host, the rodent species Apodemus agrarius, was captured. Since then numerous species in the Hantavirus genus, family Bunyaviridae, have been found, with today more than 30 species worldwide being known. Hantaviruses are hosted by rodents from the Muridae and Cricetidae families and by shrews (insectivores) in the Soricidae family. There are two types of hantavirus disease, Haemorrhagic fever with renal syndrome (HFRS) in the Old World and Hantavirus cardiopulmonary syndrome (HCPS) in the New World. The first two African hantaviruses were identified in 2006 in Guinea, West Africa; Sangassou virus (SANGV) in a rodent, the African wood mouse (Hylomyscus simus), and Tanganya virus (TGNV) in Therese’s shrew (Crocidura theresae). In this study, rodents and shrews were trapped at localities in the Western Cape and Northern Cape provinces of South Africa, and in the southern regions of Namibia. RNA was extracted from their lungs and screened for hantavirus sequences by RTPCR, using degenerate primers designed to detect all members of the Hantavirus genus. In addition, an in-house IgG ELISA assay was set up, based on recombinant N antigen from Dobrava virus, DOB-rN, and Puumala virus, PUU-rN. The assay was used to screen patient sera collected in an anonymous convenience serological survey using residual serum samples left over from routine testing at NHLS laboratories in the Western Cape for hantavirus-specific antibodies. RNA from 576 animal specimens was screened by RT-PCR; no hantavirus genome was detected in any of the specimens. Sera from 161 patients were screened for hantavirus antibodies; 11.18% of the sera were reactive to DOB-rN, 4.97% against PUU-rN and 2.48% against both antigens. v Though no virus was detected in the animals screened, this does not necessarily mean that there are no hantaviruses present in Southern Africa. A previous seroepidemiological survey conducted in South Africa reported on the presence of hantavirus specific antibodies by IFA in two species of rodents trapped in the Western Cape and Northern Cape Aethomys namquensis and Tatera leucogaster. Our was the second known study in South Africa conducted that determined and proved the presence of hantavirus specific antibodies in humans. / AFRIKAANSE OPSOMMING: Dwarsdeur die geskiedenis was daar beskrywings van siektes wat ooreenstem met die beskrywing van hantavirus simptome, maar die eerste etiologiese oorsaak van die siekte is eers in 1976 geïdentifiseer en Hantaan virus genoem, vernoem na die rivier waar naby die gasheer, Apodemus agrarius, gevang is. Van daar af het die soektog na nuwe hantavirusse intensief gevorder en vandag is daar meer as 30 spesies wêreldwyd wat aan die Hantavirus genus, ’n lid van die Bunyaviridae familie, behoort. Knaagdiere van die Muridae en Cricetidae families, sowel as spitsmuise (insekvreters) in die Soricidae familie is gasheer vir hantavirusse. Twee tipes hantavirus siekte is bekend, hemorragische koors met nier sindroom (HFRS) in die Ou Wêreld en hantavirus kardiopulmonale sindroom in die Nuwe Wêreld. Die eerste twee Afrika hantavirusse is in 2006 in Guinee Wes-Afrika geïdentifiseer; Sangassou virus (SANGV) in ’n knaagdier, die Afrika hout muis (Hylomyscus simus) en Tanganya virus (TGNV) in Therese se spitsmuis (Crocidura theresae). In hierdie studie is knaagdiere en spitsmuise op verskeie plekke in die Wes- en Noord-Kaap provinsies, asook die Suide van Namibië, gevang. RNS is onttrek vanuit die longe en hantavirus volgordes is gesoek deur middel RT-PKR deur gebruik te maak van Pan-Hanta primers wat ontwerp is om alle lede van die Hantavirus genus op te spoor. ’n Self-ontwerpde IgG ELISA, gebasseer op rekombinante N antigeen van Dobrava virus, DOB-rN en Puumala virus, PUU rN, is opgestel en gebruik om pasiënt serum, verkry in ’n anonieme serologiese opname, te toets; oorblywende serum, na toetse uitgevoer is deur NHLS laboratoriums in die Wes-Kaap, is verkry en getoets vir hantavirus spesifieke teenliggaampies. RNS van 576 dier monsters is getoets deur middel van RT-PKR en geen hantavirus is in enige van die monsters geïdentifiseer nie. Serum van 161 pasiënte is getoets vir hantavirus teenliggaampies; 11.18% van die serum was reaktief teen DOB-rN, 4.97% teen PUU-rN en 2.48% teen albei antigene. Alhoewel geen virus in die diere geïdentifiseer is nie, beteken dit nie noodwendig dat geen hantavirusse in Suidelike-Afrika voorkom nie. ‘n Vorige sero-epidemiologiese opname wat in Suid-Afrika gedoen is het die teenwoordigheid van hantavirus spesifieke teenliggaampies in twee knaagdier spesies, Aethomys namquensis en Tatera leucogaster gevang in die Wes-en Noord-Kaap, gevind. Ons studie is die tweede studie bekend in Suid-Afrika uitgevoer, wat die teenwoordigheid van hantavirus spesifieke teenliggaampies bevind en bewys het.

Ο ιός του αιμορραγικού πυρετού Κριμαίας-Κογκό (Crimean-Congo Hemorrhagic Fever Virus, CCHFV), καθώς και οι χανταϊοί (hantaviruses) προκαλούν στον άνθρωπο αιμορραγικό πυρετό. Αυτοί παρουσιάζουν ευρεία γεωγραφική κατανομή και αποτελούν απειλή για τη δημόσια υγεία, λόγω του υψηλού ποσοστού θνητότητας που σημειώνουν και της απουσίας αποτελεσματικής θεραπευτικής αγωγής. Παρότι επιδημιολογικές μελέτες δείχνουν την παρουσία αντισωμάτων στον ελληνικό πληθυσμό, περιορισμένες είναι οι αναφορές κλινικών περιστατικών CCHF και HFRS στην Ελλάδα. Σκοπός της παρούσας μελέτης είναι να προσδιορίσει τον επιπολασμό της μόλυνσης με τον CCHFV και τους χανταϊούς στον Ν. Αχαΐας, που αν και παρουσιάζει ευνοϊκές συνθήκες για την κυκλοφορία των δύο ιών, δεν έχει μελετηθεί στο παρελθόν. Σχεδιάσθηκε διατμηματική μελέτη και συγκεντρώθηκαν προοπτικά 207 δείγματα ορού φαινομενικά υγιών ατόμων-κατοίκων της περιοχής, τα οποία εξετάστηκαν με τη μέθοδο ELISA και έμμεσου ανοσοφθορισμού για την ύπαρξη αντισωμάτων έναντι του CCHFV και των χανταϊών. Ο επιπολασμός για τη μόλυνση με CCHFV βρέθηκε 3,4% και 9,7% για τη μόλυνση με χανταϊούς, ενώ κανένα από τα οροθετικά άτομα δεν ανακαλούσε συμπτώματα παρόμοια με αυτά του CCHF ή του HFRS. Για τον CCHFV, βρέθηκε ότι η ηλικία, η αγροτοκτηνοτροφική ενασχόληση, η κατοχή/εκτροφή αιγοπροβάτων, το ιστορικό νύγματος κρότωνα, η μόνιμη διαμονή σε υψόμετρο ≥400μ., η μόνιμη διαμονή σε μη αρδευόμενες αρόσιμες εκτάσεις ή σε αγροτικές εκτάσεις με σημαντικό ποσοστό φυσικής βλάστησης, καθώς και η μόνιμη διαμονή σε αγροτική περιοχή είναι σημαντικοί παράγοντες κινδύνου. Από αυτούς, το νύγμα κρότωνα, η αγροτοκτηνοτροφική ενασχόληση και η μόνιμη διαμονή σε υψόμετρο ≥400μ. βρέθηκαν να προβλέπουν καλύτερα την οροθετικότητα ενός ατόμου. Επίσης, βρέθηκε ότι παράγοντες που σχετίζονται με τη μόλυνση με χανταϊούς είναι: η ηλικία, η θέαση τρωκτικών σε ακτίνα <200μ. γύρω από την οικία και η ιδιοκτησία υπόγειας αποθήκης. Από αυτούς, μόνο η ηλικία βρέθηκε να προβλέπει καλύτερα την οροθετικότητα ενός ατόμου. Επιπλέον, παρατηρήθηκε ότι σχεδόν το 75% των θετικών ατόμων για αντισώματα έναντι των χανταϊών παρουσίαζαν ήπια επηρεασμένη νεφρική λειτουργία. Εντοπίστηκαν, επίσης, ενδημικές εστίες των ιών στον νομό: ο Δ. Ερυμάνθου για τον CCHFV και ο Δ. Δυτικής Αχαΐας για του χανταϊούς. Λαμβάνοντας υπόψη τα παραπάνω αποτελέσματα, θα πρέπει οι κλινικοί γιατροί της περιοχής να συμπεριλαμβάνουν τον CCHF και τον HFRS στη διαφορική διάγνωση εμπύρετων νοσημάτων, ιδίως όταν αυτά συνοδεύονται από θρομβοπενία ή επηρεασμένη νεφρική λειτουργία. / Crimean-Congo hemorrhagic fever virus (CCHFV) and hantaviruses cause to humans fever with hemorrhagic manifestations. These viruses present wide geographic distribution and represent major threats for public health, because of the high fatality rate that they present and the lack of appropriate treatment. Although seroprevalence studies show the presence of antibodies against CCHFV and hantaviruses in the greek population, only some reports of human cases have been reported to date in Greece. The aim of the present study is to estimate seroprevalence for CCHFV and hantaviruses in humans in the prefecture of Achaia, where the local conditions potentially favor the circulation of these viruses and which has not been previously studied. A cross-sectional study was designed and 207 human sera were collected from apparently healthy individuals living in Achaia, which were tested for CCHFV and hantaviruses IgG antibodies by ELISA and by indirect immunofluorescence assay (IFA). Seroprevalence for CCHFV infection was estimated at 3.4%, whereas for hantaviruses at 9.7%; none recalled any illness resembling CCHF or HFRS. For CCHFV, it was found that age, agro-pastoral occupation, tending sheep and/or goats, tick bite, living in areas at an altitude of ≥400m., living at rural areas, living on non-irrigated arable land or on land principally occupied by agriculture, with significant areas of natural vegetation are significantly related to seropositivity. Among them, tick bite, agro-pastoral occupation and living in areas at an altitude of ≥400m. better predict seropositivity of an individual. For hantaviruses, it was found that age, rodent sighting around home and the ownership of an underground shed are significantly related to seropositivity. Among them, it seems that only age can predict seropositivity of an individual. Moreover, it was observed that almost 75% of the seropositive for hantaviruses individuals presented mild renal dysfunction. In this study, endemic foci were also detected: the municipality of Erimanthos for CCHFV and the municipality of Western Achaia for hantaviruses. Clinicians should include CCHF and HFRS in the differential diagnosis of an acute febrile case, especially when thrombocytopenia or impaired renal function is encountered.