Made available in DSpace on 2014-12-17T14:10:27Z (GMT). No. of bitstreams: 1 CleineANM_DISSERT.pdf: 945454 bytes, checksum: 14193afcd4e4e966eebd1618875dca42 (MD5) Previous issue date: 2012-09-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Herpes simplex is a virus that can be transmitted sexually and is potentially associated with vertical transmission. This study evaluated the prevalence of genital infection by herpes simplex virus (HSV) types 1 and 2 in pregnant and nonpregnant care in the city of Natal / RN, including a total of 222 women, 92 pregnant and 130 nonpregnant. The participants answered a questionnaire to obtain data and socio-demographic characteristics, as well as potential risk factors for sexually transmitted diseases. After the interview, we collected two cervical specimens, one for the Pap test and the other for DNA extraction and analyzed by polymerase chain reaction (PCR) to detect both virus serotypes. Then the women underwent a clinical examination by colposcopy. For statistical analysis, we used the chi-square and logistic regression by SSPS 17.0 Statistic. Most women were up to 30 years of age, nonwhite ethnicity, married, elementary education, family income below the poverty level; initiated sexual activity with age up to 18 years; had more than one sexual partner lifelong and was not pregnant, but has had at least one child. The HSV-1 showed a prevalence of 26.1% among pregnant women and 30.0% in non-pregnant women. While HSV-2 prevalence was found with 10.9% and 19.2% in pregnant and nonpregnant women, respectively. The largest proportion of morphological changes of the uterine cervix was detected among nonpregnant women, both in cytology and in colposcopy. The women were nonwhite ethnicity, married, became pregnant aged less than or equal to 18 years and who had one to two pregnancies had a lower risk of acquiring genital HSV infection. There was a high prevalence of genital HSV infection, HSV-1 is more prevalent than HSV-2. No association was found between morphological changes of the uterine cervix and the presence of the virus in pregnant and nonpregnant women, nor between genital HSV infection and the classic risk factors for sexually transmitted diseases / O herpes simples ? um dos v?rus que pode ser transmitido sexualmente e est? potencialmente associado ? transmiss?o vertical. Este estudo avaliou a preval?ncia da infec??o genital pelo v?rus herpes simples (HSV) tipos 1 e 2 em mulheres gr?vidas e n?o gr?vidas atendidas no munic?pio de Natal/RN, incluindo um total de 222 mulheres, sendo 92 gr?vidas e 130 n?o gr?vidas. As participantes responderam um question?rio para a obten??o de dados e caracter?sticas s?cio-demogr?ficas, al?m de potenciais fatores de risco para doen?as sexualmente transmiss?veis. Ap?s a entrevista, foram coletados dois esp?cimes cervicais, um destinado ao exame citol?gico de Papanicolaou e o outro, para extra??o de DNA e analisado pela rea??o em cadeia da polimerase (PCR) para detectar os dois sorotipos do v?rus. Em seguida, as mulheres foram submetidas a um exame cl?nico por meio de colposcopia. Para an?lise estat?stica dos dados, utilizou-se o teste de qui-quadrado e de regress?o log?stica pelo programa SSPS Statistic 17.0. A maioria das mulheres tinha at? 30 anos de idade, etnia n?o branca, casada, grau de instru??o elementar, renda familiar de at? um sal?rio m?nimo; iniciou a atividade sexual com idade at? 18 anos; teve mais de um parceiro sexual ao longo da vida e n?o estava gr?vida, mas j? teve no m?nimo um filho. O HSV-1 apresentou preval?ncia de 26,1% nas mulheres gr?vidas e 30,0% nas n?o gr?vidas. Enquanto que o HSV-2 foi encontrado com preval?ncia de 10,9% e 19,2% nas mulheres gr?vidas e n?o gr?vidas, respectivamente. A maior propor??o de altera??es morfol?gicas da c?rvice uterina foi detectada entre as mulheres n?o gr?vidas, tanto no exame citol?gico quanto na colposcopia. As mulheres de etnia branca, casadas, que engravidaram com idade menor ou igual a 18 anos e que tiveram de uma a duas gesta??es apresentaram menor risco de adquirir infec??o genital pelo HSV. Constatou-se uma alta preval?ncia infec??o genital pelo HSV, sendo HSV-1 mais prevalente que o HSV-2. N?o se observou associa??o entre altera??es morfol?gicas da c?rvice uterina e a presen?a do v?rus nas mulheres gr?vidas e n?o gr?vidas, nem entre a infec??o genital pelo HSV e os fatores de risco cl?ssicos para doen?as sexualmente transmiss?veis

Orientador: Samir Issa Samara / Coorientador: Carlos Frederico Martins / Coorientador: José Robson Bezerra Sereno / Banca: Luis Antonio Mathias / Banca: Iveraldo dos Santos Dutra / Banca: Karen Martins Leão / Banca: Bruna Alexandrino / Resumo: Este trabalho teve como objetivo analisar parâmetros clínicos, alterações sorológicas e qualidade do sêmen de touros jovens infectados experimentalmente com o herpesvírus bovino tipo 1 (BoHV-1) entre os dias 0 e 49 pós-inoculação (PI), com o intuito de caracterizar as manisfestações que direcionam o diagnóstico, nas diversas fases de infecção. Neste experimento foram utilizados 12 touros com aproximadamente 30 meses de idade. Seis animais foram inoculados com a amostra padrão Nebraska do BoHV-1 pelas vias intranasal e intraprepucial com 10 mL de suspensão viral (107,0TCID50/mL) repartidos 5 mL em cada via. Os outros seis animais ficaram como grupo controle. Todos os touros inoculados com o BoHV-1 apresentaram edema e hiperemia no prepúcio entre o terceiro e o décimo sexto dia pi, dos quais dois também apresentaram exsudato nasal, detectado entre o terceiro e o oitavo dia PI. Nenhum dos animais do grupo controle manifestou sinais clínicos da doença. Por meio da técnica de virusneutralização (VN) foram detectados títulos de anticorpos com valores entre 4 e 512 em todos os animais inoculados, no período entre 14º e 49º dia PI. Em todos os animais inoculados foi constatada a presença de material genético do BoHV-1 nas amostras de sêmen analisadas por meio da técnica da reação em cadeia pela polimerase (PCR). Em quatro desses animais foi detectado o material genético viral aos 7,14 e 21 dias PI e em dois animais o material genético do vírus foi identificado por mais uma semana, ou seja, até o 28º dia PI. Todos os animais do grupo controle apresentaram resultados negativos nas técnicas de VN e PCR. Em relação à qualidade do sêmen, não foram observadas diferenças estatísticas (p≥0,05) entre o grupo de touros controle e o grupo de touros inoculados experimentalmente com o BoHV-1, por meio das... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study aimed to analyze clinical parameters, serological alterations and quality from the semen of young bulls experimentally infected with type 1 bovine herpes virus (BoHV-1) between days 0 and 49 post-inoculation (P. I.), in order to characterize the manifestations that orientate diagnosis, in several phases of infection. Twelve bulls were used in this experiment, all being approximately 30 months old. Six animals were inoculated with the Nebraska standard sample of BoHV-1 through intranasal and intrapreputial routes, each with 10 ml of the viral suspension (total dosage of 107.0TCID50/ml) divided in 5 ml for each route. The other six animals were used as a control group. All bulls inoculated with BoHV-1 presented edema and hyperemia on the foreskin between the third and sixteenth days after the inoculation, from which two animals also presented nasal exudate, detected between the third and eighth days P. I. No animals from the control group manifested any clinical signs characteristic of the infection caused by BoHV-1. By means of the viral neutralization (VN) technique, antibody titers with values ranging from 4 to 512 were detected in all inoculated animals between days 14 and 49 after the inoculation. In all inoculated animals, genetic material of BoHV-1 was present in semen samples analyzed by means of the polymerase chain reaction (PCR) technique. In four of these animals, viral genetic material was detected on days 7, 14 and 21 P. I. and in two animals the genetic material of the virus was identified for one more week, until day 28 after the infection. All animals in the control group showed negative results for the previously mentioned VN and PCR techniques. Considering the quality of semen samples from experimental bulls, no statistical differences were observed (p ≥ 0.05) between the control group and the group... (Complete abstract click electronic access below) / Doutor

With approximately 8.5 million unwanted cats euthanized in the U.S. annually, convenient, cost effective methods of sterilization are greatly needed. Current spay/neuter techniques, such as surgery and hormonal intervention, are not satisfying this need due to their high cost, significant expertise required, and the need for feral cats to be collected and brought into clinics for treatment. The aim of this research is to develop a safe contraceptive vaccine that could be delivered to the feral cat population in bait without compromising non-feline species. Feline Herpes Virus (FHV) is a feline specific virus. The USDA has approved the immunization of cats with an attenuated, non-pathogenic strain of FHV expressing foreign antigens. In our research, we have partially replaced Glycoprotein I of FHV to express a fusion protein of Flagellin (FliC), Enhanced Green Fluorescent Protein (EGFP), and Gonadotropin Releasing Hormone (GnRH). FliC has been shown to stimulate a heightened antibody response when antigens are expressed as fusion proteins with it. GnRH, a major reproductive hormone responsible for the development of testes and ovaries in felines, is the target of our vaccine vector. Expression of EGFP will allow tracking of the viral vector. The expression of the fusion protein (FliC-EGFP-GnRH) is expected to stimulate an antibody and cell mediated immune response directed towards feline GnRH, which will provide an immunocontraceptive effect specific to cats. / Master of Science

Recentemente, estudos têm associado a presença de vírus da família herpesviridae à doença periodontal, os quais poderiam estar envolvidos na ocorrência e progressão de diferentes formas da doença periodontal, através da supressão do sistema imune do periodonto, liberação de citotoxinas, mediadores pró-inflamatórios, o que poderia favorecer o crescimento subgengival de microrganismos. Neste estudo, testamos a hipótese de que a prevalência do herpes vírus na placa subgengival de pacientes com gengivite é igual a de pacientes portadores de periodontite crônica. Desse modo, o presente estudo teve como objetivo determinar a presença dos vírus Herpes simples vírus tipo 1 (HSV-1), Citomegalovírus (HCMV) e Epstein-Barr vírus tipo 1 (EBV-1), relacionando-os com a presença de bactérias periodontopatogênicas como: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) e Dialister pneumosintes (Dp) em amostras de placa subgengival, coletadas de 30 pacientes portadores de periodontite crônica (grupo PC), 30 pacientes com gengivite (grupo G) e 30 indivíduos periodontalmente saudáveis (grupo C). Foram coletadas quatro amostras de placa subgengival do sítio mais profundo de cada quadrante nos pacientes do grupo PC e nos pacientes do grupo G e C, um sítio aleatório de cada quadrante foi examinado. A detecção de espécies bacterianas e de herpes vírus na placa subgengival dos grupos foi realizada por PCR e Nested PCR, respectivamente. A análise estatística mostrou que o HCMV foi detectado com freqüência similar nos três grupos estudados e que houve uma maior prevalência do HSV-1, EBV-1 e P. intermedia nos pacientes com periodonite crônica e gengivite em relação ao grupo controle. Houve associação da periodontite crônica com o EBV-1 e as cinco bactérias estudadas, além da associação entre os vírus (EBV-1+ HCMV; EBV-1 + HSV-1; HSV-1 + HCMV) e entre vírus e bactérias (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans; HSV-1 + T. forsythia; HSV-1 + P. gingivalis). / Recently, studies have linked the presence of the virus family herpesviridae and periodontal disease, which may be involved in the occurrence and progression of different forms of periodontal disease through the suppression of the immune system of the periodontium, release of cytotoxins, pro-inflammatory mediators and immunopathological events, may promote growth of subgingival microorganisms. In this study, we tested the hypothesis that the prevalence of herpes virus in sub-gingival plaque of patients with gingivitis is equal to patients with chronic periodontitis. Thus, this study aimed to determine the presence of the virus Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1), relating to the presence of bacteria as periodontopathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) and Dialister pneumosintes (Dp) in subgingival plaque samples collected from 30 patients with chronic periodontitis (CP group), 30 patients with gingivitis (group G) and 30 periodontally healthy subjects (group C). We collected four samples of subgingival plaque from the deepest site in each quadrant and in the PC group and patients in group C and G, a random site in each quadrant was examined. The detection of bacterial species and herpes virus in subgingival plaque of the groups were identified by PCR and nested PCR respectively. Statistical analysis showed that HCMV was detected with a similar frequency among the three groups and significant difference in prevalence of HSV-1, EBV-1 and P. intermedia in patients with gingivitis in the control group. The chronic periodontitis was associated with EBV-1 and the five bacteria studied, and the association of the virus (EBV-1 + HCMV, EBV-1 + HSV-1, HSV-1 + HCMV) and between viruses and bacteria (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans, HSV-1 + T. forsythia; HSV-1 + P. gingivalis).

Recentemente, estudos têm associado a presença de vírus da família herpesviridae à doença periodontal, os quais poderiam estar envolvidos na ocorrência e progressão de diferentes formas da doença periodontal, através da supressão do sistema imune do periodonto, liberação de citotoxinas, mediadores pró-inflamatórios, o que poderia favorecer o crescimento subgengival de microrganismos. Neste estudo, testamos a hipótese de que a prevalência do herpes vírus na placa subgengival de pacientes com gengivite é igual a de pacientes portadores de periodontite crônica. Desse modo, o presente estudo teve como objetivo determinar a presença dos vírus Herpes simples vírus tipo 1 (HSV-1), Citomegalovírus (HCMV) e Epstein-Barr vírus tipo 1 (EBV-1), relacionando-os com a presença de bactérias periodontopatogênicas como: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) e Dialister pneumosintes (Dp) em amostras de placa subgengival, coletadas de 30 pacientes portadores de periodontite crônica (grupo PC), 30 pacientes com gengivite (grupo G) e 30 indivíduos periodontalmente saudáveis (grupo C). Foram coletadas quatro amostras de placa subgengival do sítio mais profundo de cada quadrante nos pacientes do grupo PC e nos pacientes do grupo G e C, um sítio aleatório de cada quadrante foi examinado. A detecção de espécies bacterianas e de herpes vírus na placa subgengival dos grupos foi realizada por PCR e Nested PCR, respectivamente. A análise estatística mostrou que o HCMV foi detectado com freqüência similar nos três grupos estudados e que houve uma maior prevalência do HSV-1, EBV-1 e P. intermedia nos pacientes com periodonite crônica e gengivite em relação ao grupo controle. Houve associação da periodontite crônica com o EBV-1 e as cinco bactérias estudadas, além da associação entre os vírus (EBV-1+ HCMV; EBV-1 + HSV-1; HSV-1 + HCMV) e entre vírus e bactérias (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans; HSV-1 + T. forsythia; HSV-1 + P. gingivalis). / Recently, studies have linked the presence of the virus family herpesviridae and periodontal disease, which may be involved in the occurrence and progression of different forms of periodontal disease through the suppression of the immune system of the periodontium, release of cytotoxins, pro-inflammatory mediators and immunopathological events, may promote growth of subgingival microorganisms. In this study, we tested the hypothesis that the prevalence of herpes virus in sub-gingival plaque of patients with gingivitis is equal to patients with chronic periodontitis. Thus, this study aimed to determine the presence of the virus Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1), relating to the presence of bacteria as periodontopathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) and Dialister pneumosintes (Dp) in subgingival plaque samples collected from 30 patients with chronic periodontitis (CP group), 30 patients with gingivitis (group G) and 30 periodontally healthy subjects (group C). We collected four samples of subgingival plaque from the deepest site in each quadrant and in the PC group and patients in group C and G, a random site in each quadrant was examined. The detection of bacterial species and herpes virus in subgingival plaque of the groups were identified by PCR and nested PCR respectively. Statistical analysis showed that HCMV was detected with a similar frequency among the three groups and significant difference in prevalence of HSV-1, EBV-1 and P. intermedia in patients with gingivitis in the control group. The chronic periodontitis was associated with EBV-1 and the five bacteria studied, and the association of the virus (EBV-1 + HCMV, EBV-1 + HSV-1, HSV-1 + HCMV) and between viruses and bacteria (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans, HSV-1 + T. forsythia; HSV-1 + P. gingivalis).

La dérégulation de l’expression des microARN peut induire des cancers. De plus, ils jouent un rôle crucial dans la pathogénèse et la survie des virus. L’herpès virus humain de type 8 (HHV-8 ou KSHV) est l’agent étiologique du sarcome de Kaposi et est impliqué dans la génération de lymphomes agressifs de type B. De manière intéressante, le génome ce virus code 12 pré-miARN localisés dans la région de latence et exprimés sur un même pri-miARN. Les miARN du KSHV sont importants pour le maintien de la latence, l’inhibition de l’apoptose ou encore la régulation du cycle cellulaire de l’hôte. Nous nous intéressons à leur expression et leur régulation durant l’infection virale. Nous avons résolu la structure secondaire de l’ARN codant ces miARN afin d’identifier les critères structuraux responsables de leur accumulation différentielle. Nous avons initié une analyse cinétique de la première étape de maturation et enfin nous essayons d’identifier des co-facteurs modulant leur expression. / It is now well known that modulation of microRNAs expression is linked to the development of cancers. Moreover, they play a crucial role in the pathogenesis and the survival of some viruses. Kaposi’s sarcoma associated herpes virus (KSHV) is the etiologic agent of Kaposi’s sarcoma and is involved in human aggressive B lymphomas generation. Its genome encodes 12 precursor miRNAs that are clustered in a latency region and expressed on a single long primary transcript. KSHV miRNAs are important to maintain the virus latency and to regulate or inhibit the host cell cycle or apoptosis, respectively. Therefore, understanding the regulation of KSHV miRNA accumulation is of prime importance. In this respect, we resolved the secondary structure of them iRNA cluster to identify structural criteria responsible of their differential accumulation. In addition, we started to analyse the mechanism of their maturation by kinetics studies. Finally we tried to identify some cofactors of miRNA expression.

Le Ketosis-Prone Diabetes (KPD) est un phénotype de diabète intermédiaire entre le diabète de type 1 et le diabète de type 2, fréquemment rencontré chez le sujet d’origine noire africaine. Cette forme de diabète suscite un intérêt certain de par son évolution clinique marquée notamment par la restauration de l’insulinosécrétion initialement altérée. Sobngwi et coll. en 2008 ont établi une association entre le virus HHV8 et le KPD chez des sujets Africains vivant en France. Nulle part ailleurs l’étude n’a été reproduite. L’objectif de cette thèse était de rechercher la potentielle association entre l’infection à HHV8 et le KPD ; puis d’évaluer l’impact de l’infection à HHV8 sur le profil inflammatoire des phénotypes du diabète de type 2. L’étude s’appuie sur une population de patients Africains vivant en Afrique admis consécutivement pour une décompensation hyperglycémique (glycémie à jeun≥2,5g/l) au Centre National d’Obésité de l’Hôpital Central de Yaoundé. Plus spécifiquement, il était question de :• étudier la fréquence du diabète non auto-immun à tendance cétosique (KPD); • étudier l’association entre le virus HHV8 et le KPD;• rechercher si l’infection à HHV8 est associée à un profil inflammatoire pouvant participer aux phénotypes de diabète. Etait inclus dans l’étude tout patient diabétique âgé de plus de 18 ans présentant un diabète aigu avec syndrome cardinal et cétonurie (KPD1), ceux ayant présenté un diabète inaugural aigu avec syndrome cardinal et cétonurie et en rémission depuis plus de trois mois et sans cétonurie à l’inclusion (KPD2), et ceux présentant un diabète de type 2 connu sans cétonurie (DT2). Etait exclu de l’étude tout patient présentant des stigmates d’auto-immunité du diabète de type 1 A, un diabète « MODY », une endocrinopathie, une maladie du pancréas, ou un diabète de type auto-immun. Chez l’ensemble des participants admis, nous avons collecté les données cliniques (le poids, la taille, l’IMC, le rapport tour de taille sur tour de hanche, la pression artérielle, et le pourcentage de graisse) et des prélèvements à jeun ont été effectués (sérum et cellules mononuclées du sang périphérique) pour les analyses biologiques : la glycémie par glucose oxydase, l’HbA1c par HPLC, les paramètres du profil lipidique par des methodes enzymatiques, les concentrations d’insuline et de peptide-C par électrochimiluminescence, les anticorps anti-HHV8 par immunofluorescence, l’ADN viral HHV8 par PCR en temps réel, et les marqueurs de l’inflammation par le Luminex. Les indices HOMA-β et HOMA-IR ont été utilisés pour évaluer l’insulinosécrétion et la sensibilité à l’insuline respectivement. Les marqueurs sérologiques de l’inflammation recherchés étaient : TNF-α, MCP-1, IL-8, MIP-1β, VEGF et MIP-1α... / Ketosis-Prone Diabetes (KPD) is a diabetes phenotype intermediate between type 1 and type 2 diabetes, frequently encountered in populations of African origin. This form of diabetes arouses some interest because of its clinical course marked in particular by restoring the initial impaired insulin secretion. Sobngwi et al. in 2008 established an association between HHV-8 virus and KPD in African population living in France. Nowhere else has the study been replicated. The objective of this thesis was to investigate the potential association between HHV-8 infection and KPD; then evaluate the impact of HHV-8 infection on the inflammatory profile of type 2 diabetes phenotypes. The study is based on African patients living in Africa consecutively admitted for hyperglycemic decompensation (Fasting blood glucose≥2,5g/l) at the National Obesity Centre of the Yaounde Central Hospital. More specifically, the issue was:• study the frequency of non-immune ketosis-prone diabetes (KPD);• investigate the association between HHV8 and KPD;• investigate whether HHV-8 infection is associated with an inflammatory profile that may participate in diabetes phenotypes.Was included in this study all diabetic patients old more than 18 years with acute diabetes with syndrome cardinal and ketonuria (KPD1), those who presented with an acute inaugural cardinal syndrome and diabetes ketonuria and in remission for more than three months and without ketonuria at baseline (KPD2), and those with type 2 diabetes experienced without ketonuria (T2D). Was excluded from the study all patient with stigmata of autoimmunity of type 1 A diabetes, diabetes "MODY", endocrinopathy, pancreatic disease or an autoimmune diabetes.Among all participants admitted, we collected clinical data (weight, height, BMI, waist to hip ratio, blood pressure, and the percentage of fat) and levies fasting were made (serum and peripheral blood mononuclear cells) for biological testing: glyceamia by glucose oxidase, HbA1c by HPLC, lipid profile by enzymatic methods, the insulin and C-peptide concentrations by electrochemiluminescence, anti-HHV8 antibodies by immunofluorescence, HHV8 viral DNA by real-time PCR, and markers of inflammation by Luminex. HOMA-β and HOMA-IR indices were used to assess insulinsecretion and insulinsensitivity respectively. Serological markers of inflammation investigated were : TNF-α, MCP-1, IL-8, MIP-1β, MIP-1α and VEGF...

MSc (Microbiology) / Department of Microbiology / Background: Human Herpes Virus-8 (HHV-8) and Human Immunodeficiency Virus (HIV) are endemic in sub-Saharan Africa. However, the prevalence of HHV-8 in HIV-infected individuals in sub-Saharan Africa has not been fully described and characterized. Objectives: The objective of this study was to determine the seroprevalence and genetic subtypes of HHV-8 in an HIV-infected population in Cameroon. Methodology: KSHV/HHV-8 Enzyme-linked Immunosorbent Assay (ELISA) kit (Advanced Biotechnologies Inc., USA) was used to detect IgG antibodies in the plasma of 406 HIV-infected outpatients of the Mutengene Baptist Health Centre, Cameroon. To detect the viral presence, a 233 bp fragment of the ORF 26 gene of HHV-8 was targeted by polymerase chain reaction (PCR) in total DNA purified from patients’ whole blood. A 453 bp of the K1 gene was amplified by nested PCR, sequenced and phylogenetically analysed to infer subtypes. The online tool, Synonymous Non-synonymous Analysis Program (SNAP), was used to determine the rate of synonymous and nonsynonymous mutations in the K1 gene. The genetic variability among the derived K1 nucleotide sequences was determined by mean genetic distance analysis. Results: Of the 406 participants, an HHV-8 seroprevalence of 79.1% was obtained. There was a statistically significant association of seroprevalence with age (p= 0.00), CD4+ cell count (p= 0.02), marital status (p= 0.02) and ownership of a transistor radio set (p= 0.00). Seventy samples (23.3%) were successfully amplified for ORF 26 gene confirming the presence of replicating virus. K1 sequences were obtained for 14 of the 20 (70%) K1 amplified DNAs. The mean genetic diversity of K1 sequences ranges from 0.0%-22.3%. Phylogenetic analysis revealed two infecting viral subtypes in the study cohort: subtype A5 (57.1%), and subtype B (35.7%). Greater positive selection and genetic diversity were observed in A5 subtype compared to B subtype of K1. Interestingly, one sample (BM 547) clustered with an unclassifiable sequence from South Africa. Conclusions and recommendation: This study revealed the endemicity of HHV-8 infection in the studied population, with subtypes A5 and B as the most important epidemiological genetic variants. In addition, targeting the ORF 26 region by PCR could be an approach to detect replicating virus in individuals. Further studies should investigate the association between HHV-8 infection and KS development in the study area which is endemic for HIV. This study contributes data to the HIV/HHV-8 co-infection landscape in the study area and in Africa at large.

Eine effektive Strategie zur Verhinderung der Ausbreitung von Infektionskrankheiten in der Aquakultur ist die Anwendung geeigneter Präventionsmaßnahmen, insbesondere die Impfung von Fischen. Das effektivste Impfverfahren stellt die individuelle Injektion des Impfstoffes dar. Bei Milliarden von Fischen jährlich ist dies jedoch sehr zeit- und kostenintensiv. Bei Säugetieren gilt Niederfrequenz-Sonophorese (LFS) als eine der fortschrittlichsten Technologien zur transdermalen Verabreichung von Wirkstoffen. So entstand die Idee, bei Fischen die Aufnahme von Wirkstoffen während einer Tauchbadbehandlung mit Hilfe von LFS zu verbessern. Die vorliegende Arbeit zeigt, dass mit einer Beschallungsintensität von etwa 60 mW/cm^2 die Aufnahme eines Bakterienimpfstoffs in das Kiemengewebe von Regenbogenforellen um den Faktor 240 erhöht werden konnte. Bei dieser Intensität traten geringe oder keine Nebenwirkungen auf. Bei höheren Beschallungsintensitäten wurde eine noch höhere Aufnahme des Bakterienimpfstoffes, aber auch schädliche Nebenwirkungen beobachtet. Darüber hinaus zeigte LFS eine durch eine lokale Entzündungsreaktion und Aktivierung von T-Helferzellen in den Kiemen charakterisierte, Adjuvans-ähnliche Wirkung. Ein Impfversuch mit Koi Karpfen und einem inaktivierten Impfstoff gegen das Koi-Herpesvirus zeigte, dass LFS das Potential hat, den mit einer Tauchbadimpfung erzielbaren Immunschutz zu verbessern. LFS konnte auch für die Verabreichung anderer Substanzen wie Antibiotika verwendet und die Aufnahme von Oxytetracyclin um den Faktor fünf erhöht werden. Zusammenfassend lässt sich sagen, dass die Effektivität einer Tauchbadimpfung von Fischen mit LFS durch die gesteigerte Impfstoffaufnahme und dessen Adjuvans-ähnlichen Wirkung verbessert werden kann. Darüber hinaus könnte mithilfe LFS die therapeutische Dosis von Antibiotika bei Badbehandlungen verringert werden. Es erfordert jedoch noch weitere Studien, um diese Technologie aus dem Labor in die Praxis zu übertragen. / In aquaculture, the use of prevention methods, such as vaccination of fish, is an effective strategy to avoid infectious diseases. However, the most effective route of vaccination for fish is the one-by-one intraperitoneal injection, which that is very laborious and expensive to apply for billions of fish every year. Low Frequency Sonophoresis (LFS) has been recognized as one of the most advanced technologies in transdermal delivery of substances in mammals. Thus, it has been suggested to use LFS to enhance the uptake of substances in fish during bath treatments. The present study shows that a low sonication intensity of ca. 60 mW/cm^2 at 37 kHz increased the uptake of a bacterin into the gill tissue of rainbow trout by up to a factor of 240. At this intensity, no or only minimal side effects occurred. At higher sonication intensities, an even higher bacterin uptake but also deleterious side effects were observed. In addition, LFS showed an adjuvant-like effect characterized by a local inflammatory response and T-helper cell activation in the gills. A vaccination trial with koi carp and an inactivated vaccine against the Koi Herpes Virus (KHV) showed that LFS has the potential to enhance the immune protection achieved by immersion vaccination. In addition LFS can also be used for the administration of other substances, such as antibiotics, here we showed that the uptake of Oxytetracycline could be increased by factor five. In summary, the efficacy of the immersion vaccination of fish could be improved by low-frequency ultrasound due, to the increased vaccine uptake along with its adjuvant-like effect. Furthermore, LFS could also reduce the required therapeutic dose of antibiotics in bath treatments, making them more effective, cheaper and environmentally friendly. However, further practical studies will be required to transfer this technology from the lab to the field.