Herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) are closely related viruses that establish lifelong infection in their hosts and can periodically reactivate to cause painful lesions. Approximately 50 million Americans are infected with HSV-2, the primary cause of genital ulcerative disease. In addition to the pain of the physical symptoms, genital herpes is highly stigmatized and can cause significant reductions in quality of life. HSV-2 has been demonstrated to have a synergistic relationship with HIV, where it enhances the transmissibility, susceptibility and severity of disease. In addition, potentially life-threatening neonatal herpes occurs in about 1 in 10,000 live births. While there are known means of reducing the risk of transmission, the overwhelming majority of infected persons are unaware of their status, and they remain the driving force behind new infections. Virological tests, such as PCR, are ill suited for asymptomatic infections since the detectable virus is only shed on <10% of days. Currently available type-specific serological tests based on glycoprotein G are prone to false-positive results, lack the sensitivity to detect new infections and may be affected by cross-reactivity between HSV-1 and HSV-2. Due to the known limitations of HSV serological testing, it is not recommended for the general population or pregnant women. In the first half of my thesis, I evaluate a new flow-cytometry-based method that measures serum antibody-binding to virus-infected cells (ABVIC). We obtained a panel of human control sera from Westover Heights Clinic (WHC) and determined if the ABVIC could measure HSV-specific antibody binding to test-cells. We found that the assay was sensitive, semi-quantitative, and could be made type-specific with the addition of a pre-adsorption step. The ABVIC offers an advantage over the standard of care single-antigen tests as the result measuring antibody binding to all viral proteins fixed in their native conformation. In the second half of my thesis, I used the ABVIC assay test n=34 blinded test-sera. Of these, n=17 had previously tested as “indeterminate” by bother Herpeselect ELISA as well as Western Blot. Following unblinding, we found that the ABVIC properly identified all n=17 patient sera of an unambiguous serostatus. Of the indeterminate sera, all were found to be seronegative for HSV-2. Based on these surprising results, we requested an additional n=11 indeterminate samples, which were also found to be HSV-2 seronegative. Most of the "Indeterminate" serum samples exhibited high background, which produced weak reactivity in HerpeSelect and Western blot assays, but did not confound the internally controlled ABVIC test.

Ocorrências comuns de infecções virais graves, aparecimento de cepas resistentes e um limitado número de quimioterápicos antivirais disponíveis mostram a necessidade da busca por novas substâncias ativas como antivirais. Muitas substâncias derivadas de plantas são candidatas ao estudo do seu potencial na terapia sistêmica e/ou profilaxia de herpes simplex virus 1 (HSV-1). Dentre as plantas com atividade antiviral, destacam-se as do gênero Baccharis. O objetivo geral do trabalho foi o estudo da composição química e avaliação da atividade antiviral das folhas de Baccharis anomala D.C. através de fracionamento bioguiado. Utilizando precipitação com etanol e fracionamento por permeação molecular foi possível separar os constituintes químicos ativos contra o vírus HSV-1 presentes no extrato aquoso de Baccharis anomala. Os testes de prospecção de constituintes químicos indicaram a presença de taninos, catequinas e saponinas no extrato aquoso da espécie. Através da análise cromatográfica foi possível detectar a presença de compostos fenólicos, utilizando-se coloração com cloreto férrico e reagente natural. Em relação à atividade antiviral, a fração ativa denominada AQ PPT FR 4-5 apresentou pronunciada atividade antiviral, inibindo a replicação viral em 100 % nas concentrações de 1,25, 0,625, 0,312 e 0,156 mg/mL, contra as cepas ATCC-VR733 e Aciclovir-resistente 29-R do HSV-1. Em relação ao mecanismo de ação, observou-se atividade virucida da fração AQ PPT FR 4-5. Estes resultados são muito importantes, pois, de acordo com a literatura, ainda não foram relatados compostos com atividade virucida úteis clinicamente para o tratamento de infecções por HSV-1. Conclui-se, portanto, que Baccharis anomala possui potente atividade antiviral contra o vírus HSV-1 e é promissora para estudos posteriores que visem ao isolamento, identificação e estudo do mecanismo de ação antiviral de compostos ativos da espécie, considerando a emergência de cepas resistentes e a necessidade de compostos com novos mecanismos de ação. / Common occurrences of severe viral infections, emergence of resistant strains and a limited number of available antiviral chemotherapeutics show the need to search for new active substances as antiviral. Many compounds derived from plants are candidates for the study of their potential in systemic therapy and/or prophylaxis of herpes simplex virus type 1 (HSV-1). Among the plants with antiviral activity, those from the genus Baccharis are remarkable. The main objective of the work was the study of the chemical composition and evaluation of the antiviral activity of extracts from Baccharis anomala D.C., using bioactivity guided fractionation. Through precipitation with ethanol and fractionation by molecular permeation it was achieved the separation of the active chemical constituents against HSV-1 virus in the aqueous extract of Baccharis anomala. Phytochemical tests indicated the presence of tannins, catechins and saponins in the aqueous extract. By thin layer chromatography it was detected the presence of phenolic compounds using ferric chloride and natural reagent. Concerning the antiviral activity, the active fraction named AQ PPT FR 4-5 showed pronounced antiviral activity, represented by 100 % inhibition of viral replication at concentrations of 1.25, 0.625, 0.312 and 0.156 mg/mL, against the strains ATCC-VR733 and Acyclovir-resistant 29-R of HSV-1 virus. Regarding the mechanism of action, virucidal activity on the fraction AQ PPT FR 4-5 was detected, which is also very important because, as far as we know, compounds with virucidal activity for clinical use in the treatment of HSV-1 infections were not reported yet. In conclusion, Baccharis anomala displayed pronounced antiviral activity against HSV-1 virus and it is promising for further studies aimed to the isolation, identification and mechanism of action of antiviral active compounds of the species, considering the emergence of resistant strains and the need for compounds with new mechanisms of action.

Ocorrências comuns de infecções virais graves, aparecimento de cepas resistentes e um limitado número de quimioterápicos antivirais disponíveis mostram a necessidade da busca por novas substâncias ativas como antivirais. Muitas substâncias derivadas de plantas são candidatas ao estudo do seu potencial na terapia sistêmica e/ou profilaxia de herpes simplex virus 1 (HSV-1). Dentre as plantas com atividade antiviral, destacam-se as do gênero Baccharis. O objetivo geral do trabalho foi o estudo da composição química e avaliação da atividade antiviral das folhas de Baccharis anomala D.C. através de fracionamento bioguiado. Utilizando precipitação com etanol e fracionamento por permeação molecular foi possível separar os constituintes químicos ativos contra o vírus HSV-1 presentes no extrato aquoso de Baccharis anomala. Os testes de prospecção de constituintes químicos indicaram a presença de taninos, catequinas e saponinas no extrato aquoso da espécie. Através da análise cromatográfica foi possível detectar a presença de compostos fenólicos, utilizando-se coloração com cloreto férrico e reagente natural. Em relação à atividade antiviral, a fração ativa denominada AQ PPT FR 4-5 apresentou pronunciada atividade antiviral, inibindo a replicação viral em 100 % nas concentrações de 1,25, 0,625, 0,312 e 0,156 mg/mL, contra as cepas ATCC-VR733 e Aciclovir-resistente 29-R do HSV-1. Em relação ao mecanismo de ação, observou-se atividade virucida da fração AQ PPT FR 4-5. Estes resultados são muito importantes, pois, de acordo com a literatura, ainda não foram relatados compostos com atividade virucida úteis clinicamente para o tratamento de infecções por HSV-1. Conclui-se, portanto, que Baccharis anomala possui potente atividade antiviral contra o vírus HSV-1 e é promissora para estudos posteriores que visem ao isolamento, identificação e estudo do mecanismo de ação antiviral de compostos ativos da espécie, considerando a emergência de cepas resistentes e a necessidade de compostos com novos mecanismos de ação. / Common occurrences of severe viral infections, emergence of resistant strains and a limited number of available antiviral chemotherapeutics show the need to search for new active substances as antiviral. Many compounds derived from plants are candidates for the study of their potential in systemic therapy and/or prophylaxis of herpes simplex virus type 1 (HSV-1). Among the plants with antiviral activity, those from the genus Baccharis are remarkable. The main objective of the work was the study of the chemical composition and evaluation of the antiviral activity of extracts from Baccharis anomala D.C., using bioactivity guided fractionation. Through precipitation with ethanol and fractionation by molecular permeation it was achieved the separation of the active chemical constituents against HSV-1 virus in the aqueous extract of Baccharis anomala. Phytochemical tests indicated the presence of tannins, catechins and saponins in the aqueous extract. By thin layer chromatography it was detected the presence of phenolic compounds using ferric chloride and natural reagent. Concerning the antiviral activity, the active fraction named AQ PPT FR 4-5 showed pronounced antiviral activity, represented by 100 % inhibition of viral replication at concentrations of 1.25, 0.625, 0.312 and 0.156 mg/mL, against the strains ATCC-VR733 and Acyclovir-resistant 29-R of HSV-1 virus. Regarding the mechanism of action, virucidal activity on the fraction AQ PPT FR 4-5 was detected, which is also very important because, as far as we know, compounds with virucidal activity for clinical use in the treatment of HSV-1 infections were not reported yet. In conclusion, Baccharis anomala displayed pronounced antiviral activity against HSV-1 virus and it is promising for further studies aimed to the isolation, identification and mechanism of action of antiviral active compounds of the species, considering the emergence of resistant strains and the need for compounds with new mechanisms of action.

Made available in DSpace on 2018-06-27T18:56:12Z (GMT). No. of bitstreams: 2 Larissa Daiane Willrich de Melo.pdf: 1466861 bytes, checksum: 3695d65f6aa448f284f69ac1afd4510f (MD5) Larissa Daiane Willrich de Melo.pdf.jpg: 3370 bytes, checksum: 20955982b01b119bb6af06d3b4896dcc (MD5) Previous issue date: 2013-03-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Efforts to develop an effective vaccine against Herpes Simplex Virus-1 (HSV-1) has been intense enough, however, there is no effective vaccine yet. The using of nanocapsules (Ncs) for production of viral vaccines may contribute to the enhancement and effectiveness of the already developed vaccines models. The main properties of the nanostructured system, which may contribute to the improvement of the immune response generated by vaccine is the slow and gradual release of the encapsulated content, protection of the encapsulated molecule or protein and adjuvanticity, inherent in every Ncs. The objective of this study was to produce and characterize nanocapsules with aqueous core to encapsulate the SSIEFARL peptide from HSV-1. This peptide has hydrophilic characteristics which impossibility the encapsulation in conventional nanocapsules with oil core. For the standardization of the tunnel several adaptations where modified in the system previously developed by Lambert et al. Among these, the stirring of the suspension, temperature, amount of surfactant, amount of peptide and settling time were adapted. All protocols two aqueous suspensions were tested, one with the organic solvent ethanol in the aqueous phase or only water. The results showed that the production with magnetic stirring in the presence of ethanol in the aqueous phase lead to Ncs with better physicochemical characteristics. Empty Ncs presented an average of particle size of 285 nm (+ 19,4) and zeta potential of -40.1 mV (+ 1,7) and the Ncs containing SSIEFARL peptide presented the size of 315 nm (+ 20,7) and zeta potential of -35,5 mV (+ 2,4). Other condition of production tested was the water temperature added during the production. When water was used at 40 °C the physico-chemical parameters were better than when room temperature water was used. Additionally, we tested the stability of the suspensions after storage at room temperature or 4 °C. The Ncs were stable for longer periods of time under refrigeration. For the characterization of morphostructure, Ncs were analyzed by transmission electron microscopy (TEM). This analysis revealed that Ncs present as spherical structures, homogeneous and uniform as standards in nanostructure for biological application. The analysis of Ncs by confocal microscopy was applied to investigate the encapsulation. Three amounts of SSIEFARL-6 FAM peptide were used (20, 30 or 50 &#956;L), and the 20 &#956;L showed better encapsulation. The analysis of the release profile of the peptide from Ncs showed that of the suspension produced with 20 &#956;L was more linear, demonstrating slow release for at least 24 h. These data indicate that the method of production under magnetic stirring with ethanol and 20 &#956;L of peptide in the aqueous phase is more suitable for production of aqueous core Ncs. Furthermore, empty Ncs at low concentrations showed no cytotoxicity to lymphoid cells from BALB-c in cell culture. Thus, we suggest that this approach has promising potential for developing a vaccine against HSV-1. / Os esforços para o desenvolvimento de uma vacina eficaz contra o Herpes Simplex Vírus-1 (HSV-1) tem sido bastante intensos, entretanto, ainda não existe uma vacina eficaz. A utilização de nanocápsulas (Ncs) para a produção de vacinas anti-virais pode contribuir para o aprimoramento e eficácia dos modelos vacinas já desenvolvidos. As principais propriedades desse sistema nanoestruturado, que podem contribuir para a melhora da resposta imune gerada por uma vacina são liberação lenta e gradual do conteúdo encapsulado, a proteção da molécula ou proteína encapsulada e a adjuvanticidade, inerente a cada Ncs. Assim, o objetivo deste estudo foi produzir e caracterizar nanocápsulas de núcleo aquoso, capazes de encapsular o peptídeo SSIEFARL do HSV-1. Este peptídeo possui características hidrofílicas sendo impossível a encapsulação em nanocápsulas convencionais de núcleo oleoso. Para a padronização do encapsulamento, foram combinadas diversas adaptações ao sistema previamente desenvolvido por Lambert e colaboradores. Dentre essas, foram adaptados o modo de agitação das suspensões, temperatura, quantidade de tensoativo, quantidade de peptídeo e tempo de sedimentação. Em todos os métodos foram testados duas alterações na fase aquosa das suspensões, foram produzidas Ncs com solvente etanol, na fase aquosa e/ou sem. Como resultados foram observados que a produção com agitação magnética em presença de etanol na fase aquosa foi a que produziu NCs com melhores características físico-químicas. Foram obtidas Ncs brancas com tamanho de partícula médio de 285 nm (+ 19,4) e potencial zeta de -40,1 mV (+ 1,7) e Ncs contendo o peptídeo SSIEFARL com tamanho de 315 nm (+ 20,7) e potencial zeta de -35,5 mV (+2,4). Outra condição alterada foi a temperatura da água adicionada durante os processos de produção. Quando foi utilizada água na temperatura de 40 oC os parâmetros físico-químicos foram melhores do que quando foi utilizada água na temperatura ambiente (T.A). Além disso, foi testada a estabilidade das suspensões após armazenamento em temperatura ambiente e geladeira (4oC). As nanocápsulas se mantiveram estáveis por mais tempo em sob refrigeração. Para a caracterização morfoestrutural das Ncs produzidas, foi utilizada a microscopia eletrônica de transmissão (MET). Essa análise revelou que as Ncs apresentam-se como estruturas esféricas, homogêneas e uniformes, conforme padrões estabelecidos para esse tipo de nanoestrutura. A análise das Ncs por microscopia confocal foi utilizada para comprovação do encapsulamento do peptídeo SSIEFARL-6-Fam, assim como a melhor quantidade. Foram utilizadas três quantidades de peptídeo (20, 30 ou 50 &#61549;L), sendo que a quantidade de 20 &#61549;L foi a que apresentou melhores resultados. O gráfico do perfil de liberação da suspensão produzida com 20 &#956;L mostrou-se mais linear, demonstrando liberação lenta por pelo menos 24 h. Estes dados apontam que a metodologia de produção sob agitação magnética com etanol e 20 &#956;L de peptídeo na fase aquosa apresentaram os melhores resultados para produção de nanocápsulas de núcleo aquoso. Além disso, as nanocápsulas brancas em baixas concentrações não apresentaram citotoxicidade para células de órgãos linfóides de camundongos BALB-c em cultura de células. Com isso, podemos sugerir que esta metodologia possui potencial promissor para desenvolvimento de vacina contra o HSV-1.

Submitted by MARCIA ROVADOSCHI (marciar@unifra.br) on 2018-08-16T17:48:59Z No. of bitstreams: 2 Dissertacao_LarissaDaianeWillrichDeMelo.pdf: 1512214 bytes, checksum: b264a462161b24ec3ce2aacf05afea14 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-08-16T17:48:59Z (GMT). No. of bitstreams: 2 Dissertacao_LarissaDaianeWillrichDeMelo.pdf: 1512214 bytes, checksum: b264a462161b24ec3ce2aacf05afea14 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2013-03-25 / Efforts to develop an effective vaccine against Herpes Simplex Virus-1 (HSV-1) has been intense enough, however, there is no effective vaccine yet. The using of nanocapsules (Ncs) for production of viral vaccines may contribute to the enhancement and effectiveness of the already developed vaccines models. The main properties of the nanostructured system, which may contribute to the improvement of the immune response generated by vaccine is the slow and gradual release of the encapsulated content, protection of the encapsulated molecule or protein and adjuvanticity, inherent in every Ncs. The objective of this study was to produce and characterize nanocapsules with aqueous core to encapsulate the SSIEFARL peptide from HSV-1. This peptide has hydrophilic characteristics which impossibility the encapsulation in conventional nanocapsules with oil core. For the standardization of the tunnel several adaptations where modified in the system previously developed by Lambert et al. Among these, the stirring of the suspension, temperature, amount of surfactant, amount of peptide and settling time were adapted. All protocols two aqueous suspensions were tested, one with the organic solvent ethanol in the aqueous phase or only water. The results showed that the production with magnetic stirring in the presence of ethanol in the aqueous phase lead to Ncs with better physicochemical characteristics. Empty Ncs presented an average of particle size of 285 nm (+ 19,4) and zeta potential of -40.1 mV (+ 1,7) and the Ncs containing SSIEFARL peptide presented the size of 315 nm (+ 20,7) and zeta potential of -35,5 mV (+ 2,4). Other condition of production tested was the water temperature added during the production. When water was used at 40 °C the physico-chemical parameters were better than when room temperature water was used. Additionally, we tested the stability of the suspensions after storage at room temperature or 4 °C. The Ncs were stable for longer periods of time under refrigeration. For the characterization of morphostructure, Ncs were analyzed by transmission electron microscopy (TEM). This analysis revealed that Ncs present as spherical structures, homogeneous and uniform as standards in nanostructure for biological application. The analysis of Ncs by confocal microscopy was applied to investigate the encapsulation. Three amounts of SSIEFARL-6 FAM peptide were used (20, 30 or 50 μL), and the 20 μL showed better encapsulation. The analysis of the release profile of the peptide from Ncs showed that of the suspension produced with 20 μL was more linear, demonstrating slow release for at least 24 h. These data indicate that the method of production under magnetic stirring with ethanol and 20 μL of peptide in the aqueous phase is more suitable for production of aqueous core Ncs. Furthermore, empty Ncs at low concentrations showed no cytotoxicity to lymphoid cells from BALB-c in cell culture. Thus, we suggest that this approach has promising potential for developing a vaccine against HSV-1. / Os esforços para o desenvolvimento de uma vacina eficaz contra o Herpes Simplex Vírus-1 (HSV-1) tem sido bastante intensos, entretanto, ainda não existe uma vacina eficaz. A utilização de nanocápsulas (Ncs) para a produção de vacinas anti-virais pode contribuir para o aprimoramento e eficácia dos modelos vacinas já desenvolvidos. As principais propriedades desse sistema nanoestruturado, que podem contribuir para a melhora da resposta imune gerada por uma vacina são liberação lenta e gradual do conteúdo encapsulado, a proteção da molécula ou proteína encapsulada e a adjuvanticidade, inerente a cada Ncs. Assim, o objetivo deste estudo foi produzir e caracterizar nanocápsulas de núcleo aquoso, capazes de encapsular o peptídeo SSIEFARL do HSV-1. Este peptídeo possui características hidrofílicas sendo impossível a encapsulação em nanocápsulas convencionais de núcleo oleoso. Para a padronização do encapsulamento, foram combinadas diversas adaptações ao sistema previamente desenvolvido por Lambert e colaboradores. Dentre essas, foram adaptados o modo de agitação das suspensões, temperatura, quantidade de tensoativo, quantidade de peptídeo e tempo de sedimentação. Em todos os métodos foram testados duas alterações na fase aquosa das suspensões, foram produzidas Ncs com solvente etanol, na fase aquosa e/ou sem. Como resultados foram observados que a produção com agitação magnética em presença de etanol na fase aquosa foi a que produziu NCs com melhores características físico-químicas. Foram obtidas Ncs brancas com tamanho de partícula médio de 285 nm (+ 19,4) e potencial zeta de -40,1 mV (+ 1,7) e Ncs contendo o peptídeo SSIEFARL com tamanho de 315 nm (+ 20,7) e potencial zeta de -35,5 mV (+2,4). Outra condição alterada foi a temperatura da água adicionada durante os processos de produção. Quando foi utilizada água na temperatura de 40 oC os parâmetros físico-químicos foram melhores do que quando foi utilizada água na temperatura ambiente (T.A). Além disso, foi testada a estabilidade das suspensões após armazenamento em temperatura ambiente e geladeira (4oC). As nanocápsulas se mantiveram estáveis por mais tempo em sob refrigeração. Para a caracterização morfoestrutural das Ncs produzidas, foi utilizada a microscopia eletrônica de transmissão (MET). Essa análise revelou que as Ncs apresentam-se como estruturas esféricas, homogêneas e uniformes, conforme padrões estabelecidos para esse tipo de nanoestrutura. A análise das Ncs por microscopia confocal foi utilizada para comprovação do encapsulamento do peptídeo SSIEFARL-6-Fam, assim como a melhor quantidade. Foram utilizadas três quantidades de peptídeo (20, 30 ou 50 L), sendo que a quantidade de 20 L foi a que apresentou melhores resultados. O gráfico do perfil de liberação da suspensão produzida com 20 μL mostrou-se mais linear, demonstrando liberação lenta por pelo menos 24 h. Estes dados apontam que a metodologia de produção sob agitação magnética com etanol e 20 μL de peptídeo na fase aquosa apresentaram os melhores resultados para produção de nanocápsulas de núcleo aquoso. Além disso, as nanocápsulas brancas em baixas concentrações não apresentaram citotoxicidade para células de órgãos linfóides de camundongos BALB-c em cultura de células. Com isso, podemos sugerir que esta metodologia possui potencial promissor para desenvolvimento de vacina contra o HSV-1.

of thesis Diagnostics of neuroinfection caused by human herpesviruses using nucleic acid amplification methods author: MUDr. Klára Labská supervisor: doc. MUDr. Vilma Marešová, CSc. In recent years, the diagnosis of neuroinfections has undergone a shift towards molecular biology methods. Our research focused on the predictive value of the capture of herpesvirus (HV) DNA in cerebrospinal fluid. In the first study, we examined the presence of DNA neurotropic herpes viruses (HSV1, HSV2, VZV and HHV6) in cerebrospinal fluid in immunocompetent patients with laboratory-confirmed tick-borne meningoencephalitis and enterovirus meningitis and meningoencephalitis. The control group consisted of patients with proven absence of an inflammation in the cerebrospinal fluid. Patients were followed for 6 months. The course of the disease and its consequences, including laboratory tests, were compared between groups of patients with and without the presence of HV DNA. In the second study, we tried to demonstrate the presence of HSV1 DNA in cerebrospinal fluid during its symptomatic reactivation in patients with purulent meningitis. In our group of immunocompetent patients with non-purulent inflammation in the cerebrospinal fluid, the proportion of HV DNA positive patients reached 7.5% (13 out of 173), we also...

No description available.

Purpose of Review: Recent technological developments have vastly improved our ability to study the host microbiome and its role in many disease states. Numerous other reviews have contributed to our understanding of single viruses and gut microbiota or immunological outcomes. Here, we report, in aggregate, the newest data on genital microbiota interactions with the three most common viral STIs. Recent Findings: Four themes emerge: (1) the repeatability of specific community state types corresponding with infection risk, (2) a role for the microbiota as both therapeutic target and major player in treatment efficacy, (3) a need for models in which to study the mechanisms at play in microbiota/virus interactions, and (4) the impact of microbiota populating external genitalia on viral transmission. Summary: The studies reviewed herein suggest a convoluted interplay between host microbiota and viral STIs. More mechanistic studies are needed in order to leverage these interactions to improve prevention and treatment strategies.

The purpose of this investigation was to observe changes induced by types 1 and 2 herpes simplex viruses (HSY 1 and HSY 2) when these viruses were cultivated in fish 2 cells. The possibility of attempting to use human strains of herpesviruses in order to transform normal fish tissue culture preparations was considered. Three different fish cell cultures were tested: CHSE-214 cell lines were derived from embryonic chinook salmon (Oncorhynohus tshawytscha), and STE-137 cell lines were obtained from embryonic steelhead trout (Salmo gairdneri). Cells isolated from immature ovaries of rainbow trout (Salmo gairdneri) were propagated by the investigator in these studies. Both HSV 1 and HSV 2 demonstrated cytopathic effects on the 3 different cell types described above when these cells were observed during the exponential growth phase. If virus was added before cells were dividing actively, or when the cells were maintained in a stationary phase of growth, no effect was observed. Several methods were employed in an attempt to trans- form fish cells grown in tissue culture. Different quantities of live virus, preparations of ultraviolet light in- activated virus and purified viral DNA were used in these studies but malignant conversion by known viral components was not achieved. In these experiments, the 2B strain of cells obtained originally from immature ovaries of rainbow trout appeared to undergo spontaneous transformation thereby becoming established as a permanent cell line. To date tissue cultures designated 2B have undergone 25 transfers in this laboratory.

No description available.