Preclinical assessment of an anti-HTLV-1 heterologous DNA/MVA vaccine protocol expressing a multiepitope HBZ protein.

BACKGROUND: Human T-lymphotropic virus 1 (HTLV-1) is associated with the development of several pathologies and chronic infection in humans. The inefficiency of the available treatments and the challenge in developing a protective vaccine highlight the need to produce effective immunotherapeutic tools. The HTLV-1 basic leucine zipper (bZIP) factor (HBZ) plays an important role in the HTLV-1 persistence, conferring a survival advantage to infected cells by reducing the HTLV-1 proteins expression, allowing infected cells to evade immune surveillance, and enhancing cell proliferation leading to increased proviral load. METHODS: We have generated a recombinant Modified Virus Vaccinia Ankara (MVA-HBZ) and a plasmid DNA (pcDNA3.1(+)-HBZ) expressing a multiepitope protein based on peptides of HBZ to study the immunogenic potential of this viral-derived protein in BALB/c mice model. Mice were immunized in a prime-boost heterologous protocol and their splenocytes (T CD4+ and T CD8+) were immunophenotyped by flow cytometry and the humoral response was evaluated by ELISA using HBZ protein produced in prokaryotic vector as antigen. RESULTS: T CD4+ and T CD8+ lymphocytes cells stimulated by HBZ-peptides (HBZ42-50 and HBZ157-176) showed polyfunctional double positive responses for TNF-α/IFN-γ, and TNF-α/IL-2. Moreover, T CD8+ cells presented a tendency in the activation of effector memory cells producing granzyme B (CD44+High/CD62L-Low), and the activation of Cytotoxic T Lymphocytes (CTLs) and cytotoxic responses in immunized mice were inferred through the production of granzyme B by effector memory T cells and the expression of CD107a by CD8+ T cells. The overall data is consistent with a directive and effector recall response, which may be able to operate actively in the elimination of HTLV-1-infected cells and, consequently, in the reduction of the proviral load. Sera from immunized mice, differently from those of control animals, showed IgG-anti-HBZ production by ELISA. CONCLUSIONS: Our results highlight the potential of the HBZ multiepitope protein expressed from plasmid DNA and a poxviral vector as candidates for therapeutic vaccine.

First report of colonization by Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil / Primeiro relato de colonização por Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) na Região Metropolitana de São Paulo, Brasil

Abstract We report the first known occurrence of Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil. In 2018, adult specimens were sent by residents to the competent authorities and, in the inspection of the property, a large focus associated with a marsupial nest was found. This finding reinforces the importance of the species in the state of São Paulo, serves as an alert for epidemiological surveillance and extends the species colonization area in the state of São Paulo.

Resumo Relatamos a primeira ocorrência de Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) na Região Metropolitana de São Paulo, Brasil. Em 2018, espécimes adultos foram enviados por residentes e a pesquisa na propriedade constatou a presença de um grande foco associado a um ninho marsupial. Este achado reforça a importância da espécie no estado de São Paulo, serve de alerta para a vigilância epidemiológica e amplia a área de colonização da espécie no estado de São Paulo.

Gill metazoan parasites of the spotted goatfish Pseudupeneus maculatus (Ostheichthyes: Mullidae) from the Coast of Pernambuco, northeastern Brazil / Metazoários parasitos de brânquias do saramunete Pseudupeneus maculatus (Ostheichthyes: Mullidae) da costa de Pernambuco, Nordeste do Brasil

Abstract This study evaluated the parasite fauna on the gills of spotted goatfish Pseudupeneus maculatus captured in the dry and rainy seasons in the coast of the State of Pernambuco, Brazil. Eight parasite species were identified belonging to the following taxa: Monogenea (Haliotrema caraïbensis, Haliotrema caballeroi and Haliotrema golvani); Crustacea (Rocinela signata, Hamaticolax scutigerulus and Caligidae gen. sp.) and Cestoda (Nybelinia indica and Pseudolacistorhynchus noodti). The most prevalent parasites were: Monogenea (100%), H. scutigerulus (35%), N. indica (11.7%), R. signata (8.3%), Caligidae gen. sp. (3.33%) and P. noodti (0.83%). Values of infestation are compared to other studies, and the uncommon occurrence of Trypanorhyncha on the gills suggests that the spotted goatfish could be an intermediate host for the parasite.

Resumo O presente estudo investigou a fauna parasitária das brânquias do peixe conhecido como saramunete Pseudupeneus maculatus, capturados nas estações seca e chuvosa na costa do estado de Pernambuco, Brasil. Oito espécies de parasitos foram identificadas, pertencentes aos seguintes táxons: Monogenea (Haliotrema caraïbensis, Haliotrema caballeroi e Haliotrema golvani); Crustacea (Rocinela signata, Hamaticolax scutigerulus e Caligidae gen. sp.) e Cestoda (Nybelinia indica e Pseudolacistorhynchus noodti). Os parasitos mais prevalentes foram: Monogenea (100%), H. scutigerulus (35%), N. indica (11,7%), R. signata (8,3%), Caligidae gen. sp. (3,33%) e P. noodti (0,83%). Os valores de infestação são comparados a outros estudos, e a ocorrência incomum de cestóides Trypanorhyncha sugere que o saramunete pode ser um hospedeiro intermediário para o parasito.

Impact of the anatomical location, alcoholism and smoking on the prevalence of advanced oral cancer in Brazil.

BACKGROUND: To evaluate the prevalence of oral cancer in Brazil according to the clinical stage, anatomical location, alcoholism and smoking. MATERIAL AND METHODS: Data referring to 31,217 cases of oral cancer, from 2000 to 2010, were obtained from the Integrator Module of the Hospital Registry of Cancer. Inconsistent data ("non-classified" cases) was eliminated and 21,160 cases were analyzed. The frequency distribution according to clinical stage, anatomical location, alcoholism and smoking was analyzed descriptively and through a binary logistic regression model (α<0.05). The clinical stage (dependent variable) was dichotomized in early stage (I and II) or advanced stage (III and IV). The year of diagnosis, anatomical location and deleterious habits (alcoholism and smoking) were considered independent variables. RESULTS: The most frequent characteristics were: oropharynx location (n=3856, 18.41%), clinical stage IV (n=11924, 56.09%) and combined use of alcohol and tobacco (n=19226; 61.59%). The year 2009 (p<0.01, PR = 1.162, CI-95%=1.053-1.283) and location at the base of tongue (p<0.01, PR = 2.485, CI-95% = 2.182-2.807) presented a higher prevalence ratio for advanced stage oral cancer. The combined use of alcohol and tobacco showed a higher prevalence rate for the advanced clinical stage of cancer (p<0.01, PR =1.449, CI-95%=1.382-1.520) if compared to individuals without habits, or just alcoholics. CONCLUSIONS: Higher prevalence of advanced stage of oral cancer is related to the localization at the base of the tongue and to the concomitant use of alcohol and tobacco. Therefore, it can be suggested that all these characteristics lead to a worse prognosis of oral cancer.

Efficacy of Ocimum gratissimum essential oil against the monogenean Cichlidogyrus tilapiae gill parasite of Nile tilapia / Eficácia do óleo essencial de Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae parasita de brânquias de tilápia-do-nilo

The phythotherapy is an alternative to use of chemotherapeutical agents against monogenean infection. This study evaluated the anthelmintic activity of essential oil Ocimum gratissimum against monogenean Cichlidogyrus tilapiae as well as its acute toxicity in tilapia juveniles. The mean lethal concentration (LC50) and different concentrations of the essential oil, both in vitro and in vivo assays (short and long-term baths) were assessed. The LC50 was 40.70mg.L-1 and in the in vitro assay this concentration showed 80% efficacy at the last two hours and in the in vivo assay 65.87% efficacy in long-term bath. However, it provoked morphological alterations on the gills such as hyperplasia and edema. The parasites exposure at the highest concentration (320mg.L-1) showed 100% mortality after 2h exposure in the in vitro assay, whereas in the in vivo assay, short-term baths of 5min for 3 consecutive days showed an efficacy of 87.71% without gills damage. These results demonstrate the anthelminthic activity of essential oil O. gratissimum and the safety concentration to use in Nile tilapia.(AU)

A fitoterapia é uma alternativa ao uso de agentes químicos contra infecções por parasitos monogenéticos. Este estudo avaliou a atividade anti-helmíntica do óleo essencial Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae , bem como sua toxicidade aguda e histopatologia em juvenis de tilápias. Foram avaliadas a concentração letal média (CL50) e diferentes concentrações de óleo essencial, tanto in vitro como in vivo (banho de curta e longa duração). A CL50 foi de 40,70mg.L-1; no ensaio in vitro, essa concentração apresentou 80% de eficácia, e no ensaio in vivo 65,87% de eficácia em banho de exposição crônica. No entanto, provocou alterações morfológicas nas brânquias, como hiperplasia e edema. A exposição dos parasitas na concentração mais elevada (320mg.L-1) mostrou 100% de mortalidade após duas horas de exposição no ensaio in vitro, enquanto no ensaio in vivo, em banho de curta duração de cinco minutos, durante três dias consecutivos, apresentou uma eficácia de 87,71%, sem danos às brânquias. Esses resultados demonstraram a atividade anti-helmíntica do óleo essencial de O. gratissimum e a concentração de segurança para uso na tilápia-do-nilo em banhos de curta duração.(AU)

Human leukocyte antigen distribution and genomic ancestry in Brazilian patients with sickle cell disease.

Hematopoietic stem-cell transplantation (HSCT) is currently the only established curative treatment for sickle cell disease (SCD), but is limited by donor availability. Ethnicity is thought to have an impact on the complications experienced by patients that undergo HSCT and on the likelihood of identifying an human leukocyte antigen (HLA) matched donor. In the present study, we investigated the genomic ancestry and the distribution of HLA allele groups in Brazilian patients with SCD, compared these HLA profiles to worldwide populations and evaluate the availability of HLA-matched donors. A broad intercontinental admixture of patients with SCD was observed, with African ancestry ranging from 6.7% to 93.4%. In a dendrogram based on HLA frequencies, Brazilian patients with SCD were included in a branch containing only populations with a significant African component. Among the 126 patients evaluated, 10 (8%) found a HLA-matched unrelated donor in a database of 18 134 donors. Self-reported white, brown and black matched donors were identified, and no significant difference in the percentage of compatible donors was observed between these ethnic groups. Our results show that Brazilian patients with SCD are very admixed, indicating that this group is a promising target for admixture mapping of genes involved in complications after HSCT. Additional studies may help to clarify the impact of the genetic diversity and admixture of these patients on the donor availability.

First report of colonization by Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil

Abstract We report the first known occurrence of Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil. In 2018, adult specimens were sent by residents to the competent authorities and, in the inspection of the property, a large focus associated with a marsupial nest was found. This finding reinforces the importance of the species in the state of São Paulo, serves as an alert for epidemiological surveillance and extends the species colonization area in the state of São Paulo.

Resumo Relatamos a primeira ocorrência de Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) na Região Metropolitana de São Paulo, Brasil. Em 2018, espécimes adultos foram enviados por residentes e a pesquisa na propriedade constatou a presença de um grande foco associado a um ninho marsupial. Este achado reforça a importância da espécie no estado de São Paulo, serve de alerta para a vigilância epidemiológica e amplia a área de colonização da espécie no estado de São Paulo.

A new species of Tripartiella (Ciliophora: Trichodinidae) from Aequidens tetramerus (Perciformes: Cichlidae) in north Brazil / Uma nova espécie de Tripartiella (Ciliophora: Trichodinidae) em Aequidens tetramerus no norte do Brasil

Abstract A new species of Tripartiella is described from the gills of the wild saddle cichlid Aequidens tetramerus in north Brazil. Wet smears of skin and gills of examined fish were air-dried at room temperature and impregnated with Klein’s dry silver method for examination of the adhesive disc’s structures and denticles. Total prevalence of parasitism was 65%. This ciliate is characterized as a small-sized trichodinid, body diameter 37.03 ± 4.9 μm, adhesive disc 30.50 ± 2.71 μm, denticulate ring 13.28 ± 0.8 μm and 24 ± 2.0 denticles. Taxonomic and morphometric data for the new species are discussed.

Resumo Um nova espécie de Tripartiella encontrada nas brânquias de Aequidens tetramerus da região Norte do Brasil é descrita. Esfregaços de pele e brânquias de peixes examinados foram secos à temperatura ambiente e impregnados pelo método de Klein para observação das estruturas do disco adesivo e dentículos. A prevalência do parasitismo foi de 65%. Este ciliado é caracterizado como pequeno trichodinídeo, com corpo medindo 37,03 ± 4,9 μm de diâmetro, disco adesivo com 30,50 ± 2,71 μm, anel denticulado com 13,28 ± 0,8 μm e 24 ± 2,0 dentículos. Dados taxonômicos e morfométricos para a nova espécie são discutidos.

The role of TWEAK/Fn14 signaling in the MPTP-model of Parkinson's disease.

The tumor necrosis factor like weak inducer of apoptosis (TWEAK) and its receptor, fibroblast growth factor-inducible 14 (Fn14), mediate inflammation and neuronal apoptosis in cerebral edema, ischemic stroke and multiple sclerosis. The downstream effectors and pathways linked to TWEAK-Fn14 signaling are strongly implicated in the pathology of Parkinson's disease (PD), thus indicating a putative role for TWEAK/Fn14 signaling in PD neurodegeneration. Using the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model, we aimed to determine whether genetic ablation or pharmacologic mitigation of the TWEAK protein and its Fn14 receptor affected substantia nigra and striatum Parkinsonian pathology. Changes in endogenous TWEAK protein expression were also quantified in tissue from both MPTP-treated mice and PD human samples. TWEAK protein expression was transiently increased in the striatal tissue but remained unaltered in substantia nigra tissue of MPTP-treated mice. There was also no change of TWEAK protein levels in the substantia nigra or the striatum of human PD patients as compared to matched control subjects. Mitigating the effects of endogenous TWEAK protein using neutralizing antibody did affect MPTP-mediated neurotoxicity in the substantia nigra using the sub-acute model of MPTP (30mg/kg i.p. over five consecutive days). Neither TWEAK nor Fn14 genetic ablation led to attenuation of MPTP-toxicity in the acute model. These findings suggest that TWEAK signaling might be an aspect of MPTP-mediated neuropathology and be involved in the overall neurodegenerative pathology of PD.

First record of larvae of Hysterothylacium (Nematoda: Anisakidae) with zoonotic potential in the pirarucu Arapaima gigas (Osteichthyes: Arapaimidae) from South America / Primeiro registro de larvas de Hysterothylacium sp.(Nematoda: Anisakidae) com potencial zoonótico em pirarucu Arapaima gigas (Osteichthyes: Arapaimidae) na América do Sul

Abstract Third-stage larvae (L3) of Hysterothylacium sp. were collected by the first time in juveniles of pirarucu Arapaima gigas farmed in the Rio Preto da Eva, Amazonas state. Ninety-eight (98) out of 100 examined fish showed to be parasitized. Five hundred and ninety larvae of Hysterothylacium sp. were collected from the intestines, stomach and pyloric caeca. The mean intensity of parasite indexes was 6.02 (±5.75) ranging from 1 to 40 larvae per host and the mean abundance was 5.9 (±5.76). The A. gigas is the new host record for larvae of Hysterothylacium sp. in Brazil, and this is the first record of larvae of Hysterothylacium (Nematoda: Anisakidae) with zoonotic potential in the pirarucu from South America.

Resumo Larvas de terceiro estágio (L3) de Hysterothylacium sp. foram coletadas pela primeira vez em juvenis de pirarucu Arapaima gigas cultivados no Rio Preto da Eva, Estado do Amazonas. Noventa e oito (98) dos 100 peixes examinados estavam parasitados. Quinhentos e noventa larvas de Hysterothylaciumsp.foram coletados no intestino, estômago e cecos pilóricos. O índice parasitário de intensidade média foi de 6,02 (±5,75) variando de 1 a 40 larvas por hospedeiro e o de abundância média foi de 5,9 (±5,76). A. Gigas é um novo registro de hospedeiro para larvas de Hysterothylacium sp. no Brasil, também sendo, o primeiro registro de larvas de Hysterothylacium sp. com potencial zoonótico em pirarucu da América do Sul.

Effect of water temperature and salinity in oviposition, hatching success and infestation of Aphanoblastella mastigatus (Monogenea, Dactylogyridae) on Rhamdia quelen / Efeito da temperatura da água e salinidade na oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus (Monogenea, Dactylogyridae) em Rhamdia quelen

Abstract Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasite’s survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatus’s eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Resumo Diversos parâmetros ambientais podem afetar os processos biológicos de diversos organismos invertebrados aquáticos, como os Monogenea. Neste estudo, nós investigamos a oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus, parasito de jundiá Rhamdia quelen, em diferentes condições de temperatura (~ 24 e 28 °C) e salinidade (por meio da adição de cloreto de sódio na água nas concentrações de 0, 5 e 9 g/L) em laboratório. Não houve diferença significativa na taxa de oviposição nem no sucesso de infestação de A. mastigatus à 24 e 28 °C; por outro lado, a concentração de 9 g/L de cloreto de sódio na água inviabilizou não só a sobrevivência do parasito mas também a viabilidade dos ovos. Esse resultado indica que seu uso é eficiente como potencial tratamento profilático. A taxa de eclosão dos ovos de A. mastigatus foi significativamente maior a 28 °C. A partir de 5 g/L, a mesma foi significativamente menor. Durante um período de 24 horas, dois picos na oviposição ocorreram nos momentos de maior variação da luminosidade (06h15 e 18h15). Futuros estudos são recomendados utilizando intervalos de temperatura e intensidades de infestação experimental maiores para verificar a influência da temperatura na longevidade e sucesso de infestação de A. mastigatus.

Effect of water temperature and salinity in oviposition, hatching success and infestation of Aphanoblastella mastigatus (Monogenea, Dactylogyridae) on Rhamdia quelen

Abstract Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasites survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatuss eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Resumo Diversos parâmetros ambientais podem afetar os processos biológicos de diversos organismos invertebrados aquáticos, como os Monogenea. Neste estudo, nós investigamos a oviposição, sucesso de eclosão e infestação de Aphanoblastella mastigatus, parasito de jundiá Rhamdia quelen, em diferentes condições de temperatura (~ 24 e 28 °C) e salinidade (por meio da adição de cloreto de sódio na água nas concentrações de 0, 5 e 9 g/L) em laboratório. Não houve diferença significativa na taxa de oviposição nem no sucesso de infestação de A. mastigatus à 24 e 28 °C; por outro lado, a concentração de 9 g/L de cloreto de sódio na água inviabilizou não só a sobrevivência do parasito mas também a viabilidade dos ovos. Esse resultado indica que seu uso é eficiente como potencial tratamento profilático. A taxa de eclosão dos ovos de A. mastigatus foi significativamente maior a 28 °C. A partir de 5 g/L, a mesma foi significativamente menor. Durante um período de 24 horas, dois picos na oviposição ocorreram nos momentos de maior variação da luminosidade (06h15 e 18h15). Futuros estudos são recomendados utilizando intervalos de temperatura e intensidades de infestação experimental maiores para verificar a influência da temperatura na longevidade e sucesso de infestação de A. mastigatus.

Helicometrina nimia Linton, 1910 (Digenea: Opecoelidae) in dusky grouper Epinephelus marginatus (Lowe, 1834) (Teleostei: Serranidae) from southeastern Brazil / Helicometrina nimia Linton, 1910 (Digenea: Opecoelidae) em garoupa verdadeira Epinephelus marginatus (Lowe, 1834) (Teleostei: Serranidae) no Sudeste do Brasil

Helicometrina nimia (Opecoelidae) is a digenean with wide distribution. Fish families most commonly used as hosts for H. nimia are Serranidae, Pomodasydae, Scorpaenidae and Clinidae. In the present study, a new host and a new host locality are presented for the species. A description of the studied specimens, besides comments concerning its taxonomic status and biometrically compared tables of H. nimia reports are given. The taxonomic status of members of Helicometrina has been questionable. The greatest controversy for the genus seems to be related to the validity of diagnostic features, especially in regard to the number of testes. In the present study, all studied specimens presented a permanent and steady number of testes (n=9) and therefore its use as a diagnostic character is supported by the present authors. Epinephelus marginatus is considered a new host for Helicometrina nimia, and São Paulo state, southeastern Brazil, a new locality for the species.

Helicometrina nimia (Opecoelidae) é um digenético com distribuição mundial. As famílias de peixes mais comumente utilizadas como hospedeiros por H. nimia são Serranidae, Pomodasydae, Scorpaenidae e Clinidae. No presente estudo, um novo hospedeiro e uma nova localidade são apresentados para a espécie. A descrição dos espécimes estudados, além de comentários sobre o seu status taxonômico e uma tabela comparativa de medidas de H. nimia são apresentados. O status taxonômico dos membros de Helicometrina tem sido questionável. A maior controvérsia dentro do gênero parece estar relacionada com a validade das características diagnósticas, especialmente no que diz respeito ao número de testículos. No presente estudo, todos os espécimes estudados apresentaram número permanente e estável de testículos (n=9) e, portanto, sua utilização como caráter diagnóstico é suportada pelos presentes autores. Epinephelus marginatus é considerada um novo hospedeiro para Helicometrina nimia e São Paulo, sudeste do Brasil, uma nova localidade para a espécie.

Parasite fauna of wild and cultured dusky-grouper Epinephelus marginatus (Lowe, 1834) from Ubatuba, southeastern Brazil / Fauna parasitária da garoupa selvagem e de cultivo Epinephelus marginatus (Lowe, 1834) em Ubatuba, sudeste do Brasil

This study aimed at identifying and quantifying the parasites of wild and cultured dusky grouper Epinephelus marginatus. During a year and thereby all four seasons, 20 wild and 20 cultured groupers were examined for the presence of parasites, except in the last season, in which 19 wild and 20 cultured fish were examined, totalling 159 groupers analysed from Ubatuba, southeastern Brazil. Prevalence, mean intensity of infection, mean abundance and mean relative dominance were calculated. Five species of parasites were identified in fish from both origins: Pseudorhabdosynochus beverleyburtonae (Monogenea), Neobenedenia melleni (Monogenea), Pseudempleurosoma sp. (Monogenea), Helicometrina nimia (Digenea) and larvae of Contracaecum sp. (Nematoda). The prevalence of ectoparasites, in most cases, was higher than endoparasites. The most abundant parasite was the monogenea Pseudorhabdosynochus beverleyburtonae in both wild and cultured fish, along all seasons. Neobenedenia melleni was observed in wild and cultured fish in all seasons, with a gradual increase in the number of parasites from the coldest to the hottest seasons, with the highest prevalence and mean intensity in the summer. Helicometrina nimia was found in all seasons in both wild and cultured fish, except for summer, where its presence was detected only in wild fish. Pseudempleurosoma sp. and larvae of Contracaecum sp. showed low prevalence occurring in wild and cultured fish in the autumn and spring, respectively. This study revealed high intensities of potentially pathogenic parasites that could favour disease outbreaks in culture conditions.

Este estudo teve como objetivo identificar e quantificar os parasitos da garoupa verdadeira Epinephelus marginatus selvagem e cultivada. Durante um ano, nas quatro estações, 20 garoupas selvagens e 20 cultivadas foram examinadas quanto à presença de parasitos, exceto na última estação, em que foram examinados 19 peixes selvagens e 20 cultivados, totalizando 159 peixes examinados em Ubatuba, Sudeste do Brasil. A prevalência, intensidade média de infecção, abundância média and dominância média relativa foram calculados. Cinco espécies de parasitos foram identificadas nos peixes de ambas as origens: Pseudorhabdosynochus beverleyburtonae (Monogenea), Neobenedenia melleni (Monogenea), Pseudempleurosoma sp. (Monogenea), Helicometrina nimia (Digenea) e larvas de Contracaecum sp. (Nematoda). A prevalência de ectoparasitos foi, na maioria das vezes, maior do que a de endoparasitos. O parasito mais abundante foi o monogenea Pseudorhabdosynochus beverleyburtonae em ambos os peixes selvagens e cultivados, durante todas as estações. Neobenedenia melleni foi observado e, peixes selvagens e cultivados em todas as estações, com aumento gradativo no número de parasitos das estações mais frias para as mais quentes, com as maiores prevalências e intensidades médias no verão. Helicometrina nimia foi encontrado em todas as estações em ambos os peixes selvagens e cultivados, exceto no verão, em que foi observada sua presença apenas em peixes selvagens. Pseudempleurosoma sp. e larvas de Contracaecum sp. apresentaram baixas prevalências, ocorrendo em peixes selvagens e cultivados no outono e na primavera, respectivamente. Este estudo revelou altas intensidades de parasitos potencialmente patogênicos que podem favorecer surtos de doenças em situações de cultivo.

Questing for an optimal, universal viral agent for oncolytic virotherapy.

One of the most promising strategies to treat cancer is attacking it with viruses designed to exploit specific altered pathways. Here, the effects of oncolytic virotherapy on tumors having compact, papillary, and disconnected morphologies are investigated through computer simulations of a multiscale model coupling macroscopic reaction-diffusion equations for the nutrients with microscopic stochastic rules for the actions of individual cells and viruses. The interaction among viruses and tumor cells involves cell infection, intracellular virus replication, and the release of new viruses in the tissue after cell lysis. The evolution over time of both the viral load and cancer cell population, as well as the probabilities for tumor eradication, were evaluated for a range of multiplicities of infection, viral entries, and burst sizes. It was found that in immunosuppressed hosts, the antitumor efficacy of a virus is primarily determined by its entry efficiency, its replicative capacity within the tumor, and its ability to spread over the tissue. However, the optimal traits for oncolytic viruses depend critically on the tumor growth dynamics and do not necessarily include rapid replication, cytolysis, or spreading, currently assumed as necessary conditions for a successful therapeutic outcome. Our findings have potential implications on the design of new vectors for the viral therapy of cancer.

Laboratorial diagnosis of paracoccidioidomycosis and new insights for the future of fungal diagnosis.

Paracoccidioidomycosis (PCM) is the most prevalent mycosis in Latin-America. As for other mycosis, its importance of has been largely underestimated, partially due to the limited geographical distribution of the etiologic fungal agent (Paracoccidioides brasiliensis). However, the advent of AIDS and other immune suppressing conditions is creating an emergent need for improved diagnostic tests envisaging simpler, cheaper, faster and more sensitive and accurate detection of pathogenic fungi, especially those causing systemic and opportunistic diseases. Routine laboratorial diagnosis of PCM disease relies mainly on direct observation of the fungus. However, culture growing is slow and, too often, definite diagnosis can only be obtained at later growing stages. Immunodiagnosis is also widely employed, although usually cumbersome and complex. Enzyme-based immunoassays are more amenable to automation for high-throughput testing, but may lead to cross-reactivity with other fungi. Plus, molecular diagnosis relying on polymerase-chain reaction (PCR) and nucleic-acid hybridization, although still at early stages of application to routine diagnosis of P. brasiliensis, has triggered the development of techniques for its improved specific detection, thus contributing for epidemiological studies as well. In the future, microarrays and newer biosensing technologies, coupled to new bionanotechnological tools, will certainly improve diagnosis of PCM and other mycosis through very specific and sensitive pathogen biomolecular detection.

Infecção por cestóides proteocefalídeos em tucunaré Cichla sp. (Osteichthyes: Cichlidae), no Rio Paraná, São Paulo / Proteocephalid cestode infection in tucunaré Cichla sp. (Osteichthyes: Cichlidae) from Paraná River, São Paulo

The occurrence of proteocephalid cestodes in tucunaré Cichla sp., captured monthly, between August 2000 and August 2001, in Paraná River, Presidente Epitácio, SP, was evaluated. From 128 specimens, 71 (55.6 percent) were parasitized by Proteocephalus macrophallus (Diesing, 1850) and/or P. microscopicus (Woodland, 1935). Total mean abundance and intensity were 157.08 and 223.41, respectively. The highest prevalence (90 percent) mean abundance (1,122.4) and intensity indexes (1,247.11) occurred in February 2001, while in September 2000 there were no observed animals infected by cestodes. No relationship between the sex of the host and parasitological indexes was found.

Avaliou-se a ocorrência de cestóides proteocefalídeos em tucunaré Cichla sp., capturados mensalmente, entre agosto de 2000 e agosto de 2001, no rio Paraná, em Presidente Epitácio, SP. Um total de 128 espécimes foram analisados, dos quais 71 (55,6 por cento) estavam parasitados por Proteocephalus macrophallus (Diesing, 1850) e/ou P. microscopicus (Woodland, 1935). A abundância e intensidade média total foram de 157,08 e 223,41, respectivamente. A maior prevalência (90 por cento), juntamente com os maiores índices de abundância (1122,4) e intensidade média (1247,11) ocorreram no mês de fevereiro 2001, enquanto no mês de setembro 2000 não foram observados animais parasitados por cestóides. Não houve relação entre o sexo do hospedeiro e os índices parasitológicos.

First record of Trichodina heterodentata (Ciliophora: Trichodinidae) from channel catfish, Ictalurus punctatus cultivated in Brazil / Primeiro registro de Trichodina heterodentata (Ciliophora: Trichodinidae) em bagre-do-canal, Ictalurus punctatus cultivado no Brasil

This study characterises morphologically Trichodina heterodentata Duncan, 1977 from channel catfish, Ictalurus punctatus (Rafinesque, 1818) in the State of Santa Catarina, Brazil. Body and gill smears were air-dried at room temperature, impregnated with silver nitrate and/or stained with Giemsa. Ten characteristics were selected to compare the present material with other morphological characterisations of T. heterodentata. Prevalence rate was 100 percent, mean intensity 89,333.70 (3,125 to 299,100 parasites per host). Trichodina heterodentata was considered medium-sized trichodinid with mean body diameter 59.4 ± 8.5 μm, denticulate ring 38.5 ± 4.5 μm, adhesive disc 60.2 ± 6.7 μm diameter and 24.4 ± 1.6 denticles. In relation to previous reports of T. heterodentata this material resembles in 90 percent of the analysed characters. This work confirms the biometrical variation that exists in the different populations of T. heterodentata. A list of hosts and comparative measurements of T. heterodentata are presented and the channel catfish is considered a new host.

Este estudo caracteriza morfologicamente Trichodina heterodentata Duncan, 1977 em bagre-do-canal, Ictalurus punctatus (Rafinesque, 1818) no Estado de Santa Catarina, Brasil. Esfregaços do corpo e brânquias foram secados à temperatura ambiente, impregnados com nitrato de Prata e/ou corados com Giemsa. Dez características foram selecionadas para comparar o presente material com as diferentes caracterizações morfológicas de T. heterodentata. A taxa de prevalência foi de 100 por cento, a intensidade média foi de 89.333.75 (3.125 a 299.100 parasitos por hospedeiro). Trichodina heterodentata foi considerado um tricodinídeo de tamanho médio com a média do diâmetro do corpo de 59,4 ± 8.5 μm, anel denticulado 38,5 ± 4,5 μm, disco adesivo 60,2 ± 6,7 μm de diâmetro e 24,4 ± 1,6 dentículos. Em relação a registros prévios de T. heterodentata, 90 por cento das características foram semelhantes. Este trabalho confirma a variação biométrica que existe em diferentes populações de T. heterodentata. Uma lista de hospedeiros e medidas comparativas de T. heterodentata são apresentadas e o bagre-do-canal considerado um novo hospedeiro.

Effect of probiotic supplemented diet on marine shrimp survival after challenge with vibrio harveyi / Efeito do uso de dieta suplementada com o probiótico sobre a sobrevivência de camarões marinhos após o desafio com vibrio harveyi

The effect of a Lactobacillus plantarum-supplemented diet on shrimp growth, digestive tract bacterial microbiota, survival, and some hemato-immunological parameters after an experimental challenge with Vibrio harveyi was studied. No difference (P>0.05) was observed in shrimp survival (80±6 percent) and final weight (6.63±0.56g) after 60 days feeding trial. Total bacteria count and Vibrio spp. count in the digestive tract were not diferent (P>0.05) until day 40, but they were lower (P<0.05) in the probiotic-supplemented group on day 60. Total lactic bacteria in the shrimp digestive tract was higher after day 20 in the probiotic-supplemented group. Ten hours after V. harveyi challenge, survival of the probiotic-supplemented group (65.7±2.9 percent) was higher (P>0.05) than the control group (39.9±4.4 percent). Bacterial counts in hemolymph and hepatopancreas was lower (P<0.05) in the probiotic-supplemented group than in the control group after V. harveyi challenge. Total hemocyte count and serum agglutination activity were higher (P>0.05) in the probiotic-supplemented group after challenge with V. harveyi. Probiotic-supplemented diet modifies shrimp digestive tract bacterial microbiota, increasing resistance to V. harveyi infection.

Avaliou-se o efeito de uma dieta suplementada com Lactobacillus plantarum na engorda de Litopenaeus vannamei sobre a microbiota bacteriana do trato digestivo, a sobrevivência e alguns parâmetros hematoimunológicos após desafio com Vibrio harveyi. Não foi observada diferença significativa na sobrevivência (80±6 por cento) e no peso final dos camarões (6,63±0,56g) durante a engorda (60 dias). A população de bactérias totais e Vibrio spp. no intestino não diferiu até o 40º dia de cultivo, mas foi inferior nos camarões alimentados com dieta suplementada com probiótico no 60º dia de cultivo. A população de bactérias lácticas foi superior no intestino de camarões alimentados com dieta suplementada com probióticos após o 20º dia de cultivo. Dez horas após o desafio, a sobrevivência dos camarões alimentados com dieta suplementada com probiótico foi superior (65,7±2,9 por cento) a dos camarões do controle (39,9±4,4 por cento). Camarões alimentados com dieta suplementada com probióticos e desafiados com V. harveyi apresentaram menor contagem de bactérias na hemolinfa e hepatopâncreas. A contagem total de hemócitos e a atividade aglutinante da hemolinfa foram superiores nos camarões inoculados com V. harveyi alimentados com dieta suplementada com probióticos. A suplementação da dieta com probióticos modifica a microbiota bacteriana intestinal dos camarões aumentando sua resistência ao desafio com V. harveyi.

Lookback study of HTLV-1 and 2 seropositive donors and their recipients in Belo Horizonte, Brazil.

The objective of this study was to perform lookback study in recipients of blood components from human T-lymphotropic virus (HTLV) seropositive donors. HTLV-1/2 may be transmitted by blood transfusion. Brazil is an endemic area for the virus and its screening in blood donors is mandatory since 1993. Hemominas Foundation (HF) is the public transfusion centre in Minas Gerais, Brazil. Data on HTLV-1/2 seropositive donors and recipients from 1993 to 2004 were obtained at HF and 24 contracting hospitals. From 1993 to 2004, HTLV-1/2 enzyme immunoassay (EIA) was performed in 918 678 donations of approximately 422 600 blood donor candidates. Of these, 456 donors (0.1%) were reactive and confirmed by Western blot (WB): 449 HTLV-1 and 7 HTLV-2. Sixty-six (14.5%) were repeat donors and had 194 blood cellular components produced from their previous donations. Of the distributed components, 119/146 (81.5%) had the recipient traced, with a total of 114 individuals. Of these, only 13 recipients were tested: six (46%) were HTLV-1 positive (four recipients of red cell units, two of platelets) and seven (54%) were negative (six of red cell units and one of platelets). Eleven did not respond and 62/114 (54.0%) were deceased. Another 28/114 (25.0%) could not be located. All six seropositive HTLV-1 recipients identified had no symptoms suggestive of HTLV-1-associated diseases. Acellular components, when used alone, were not associated with HTLV seropositivity. HTLV-1 transmission by cellular blood components occurred before screening for the virus was introduced. Haemovigilance was difficult to perform due to unavailability of computer systems before 1999 and to inadequate medical records at hospitals.