Isolation of bovine coronavirus (BCoV) in monolayers of HmLu-1 cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histopathological and ultrastructural characteristics of myeloid leukosis in broiler chicken

Caracterizaram-se a linhagem e o grau de diferenciação das células neoplásicas no estudo histopatológico e ultraestrutural da leucose mielóide. Histologicamente as células neoplásicas apresentaram pleomorfismo, núcleos ovais, nucléolos proeminentes, cromatina distribuída de maneira irregular, figuras de mitose atípicas e moderada quantidade de citoplasma contendo granulações eosinofílicas esféricas. Essas características indicam a linhagem mielóide. Ultraestruturalmente evidenciaram-se células com núcleo oval, volumoso, eletrodenso, com predomínio de eucromatina e citoplasma com numerosos grânulos esféricos, eletrodensos e homogêneos, indicando mielócitos com diferenciação para eosinófilos. Constatou-se também a presença de partículas virais tipo-C no espaço intercelular dos túbulos renais, no interior de vesículas intracitoplasmáticas dos mielócitos imaturos presentes na medula óssea e ovário, e PCR positivo para ALV-J.

In vitro evaluation of three different biomaterials as scaffolds for canine mesenchymal stem cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação de uma equação de predição das exigências protéicas para aves reprodutoras pesadas na fase de produção

O objetivo do presente trabalho foi avaliar uma equação de predição das exigências de proteína bruta (PB) para reprodutoras pesadas na fase de produção. O experimento foi realizado com 600 aves reprodutoras pesadas, Hubbard HI-Y, durante o período de 31 a 46 semanas de idade, alojadas em boxes num delineamento experimental inteiramente casualizado com três tratamentos e cinco repetições de 40 aves. Os tratamentos consistiram de: T1- Fornecimento de PB de acordo com o manual da linhagem (controle), T2- Fornecimento de PB de acordo com a equação de predição determinada, utilizando os dados de desempenho médio das aves do tratamento controle para predizer as exigências e T3- Fornecimento de PB de acordo com a equação de predição determinada, utilizando os dados de desempenho de cada parcela experimental para predizer as exigências, onde a equação de predição avaliada foi: PB=2,282.P0,75+0,356.G+0,262.MO, sendo PB a exigência de proteína bruta (g/ave/dia), P o peso corporal (kg), G o ganho de peso (g) e MO a massa de ovos (g). As rações foram formuladas para atender as exigências nutricionais e quando necessário eram incluídos os aminoácidos sintéticos, metionina, lisina, triptofano, treonina e arginina. As aves alimentadas de acordo com a equação ingeriram menores quantidades de proteína (20,8g/dia) quando comparadas às alimentadas de acordo com as recomendações (23,80g), entretanto isto levou a menores pesos dos ovos refletindo no peso dos pintos. A equação de predição proporcionou melhores resultados quanto à eficiência protéica. Assim, concluiu-se que a equação de predição não forneceu a quantidade mínima de proteína bruta para atender as exigências dos aminoácidos não suplementados na dieta.

Identificação da Leptospira interrogans sorovar pomona em camundongos geneticamente selecionados, para a alta e baixa produção de anticorpos, através da técnica de imunoperoxidase em tecido renal e isolamento bacteriano em meio de Fletcher

O presente trabalho teve por objetivo identificar a presença da Leptospira interrogans sorovar pomona em camundongos geneticamente selecionados para a alta e baixa resposta a anticorpos. Todos os animais foram submetidos ao isolamento bacteriano, imunohistoquímica (imunoperoxidase) em cortes de tecido renal e coloração através da hematoxilina-eosina. A técnica de imunoperoxidase apresentou-se pouco mais sensível em relação ao cultivo, entretanto, ambas foram bons parâmetros de identificação do agente. Presença de lesões renais mais intensas ocorreram em períodos em que houve maior número de bactérias isoladas em meio de cultivo. Camundongos da linhagem HIV-A conseguiram eliminar as leptospiras com maior eficiência e rapidez em relação as linhagem LIV-A, entretanto o estudo demonstrou que ambas linhagens da seleção IV-A foram eficientes em controlar o processo infeccioso.

Mecanismos celulares e moleculares que controlam o desenvolvimento e o crescimento muscular

O músculo estriado esquelético é formado pela associação de fibras musculares com a matriz extracelular. Esse tecido possui alta plasticidade e o conhecimento das características morfológicas, da miogênese, e da dinâmica do crescimento é importante para o entendimento da morfofisiologia bem como para a seleção de animais visando a melhoria na produção de carne. A maioria dos músculos estriados originam-se de células precursoras do mesoderma a partir dos somitos do embrião e o controle da diferenciação ocorre pela ação de fatores indutores ou inibidores. Um grupo de fatores transcricionais, pertencentes à família MyoD tem um papel central na diferenciação muscular. Coletivamente chamados de Fatores de Regulação Miogênica (MRFs), são conhecidos quatro tipos: MyoD, myf-5, miogenina e MRF4. Esses fatores ligam-se à seqüências de DNA conhecidas como Ebox (CANNTG) na região promotora de vários genes músculo-específicos, levando à expressão dos mesmos. As células embrionárias com potencial para diferenciação em células musculares (células precursoras miogênicas) expressam MyoD e Myf-5 e são denominadas de mioblastos. Essas células proliferam, saem do ciclo celular, expressam miogenina e MRF4, que regulam a fusão e a diferenciação da fibra muscular. Uma população de mioblastos que se diferencia mais tardiamente, as células miossatélites, são responsáveis pelo crescimento muscular no período pós natal, que pode ocorrer por hiperplasia e hipertrofia das fibras. As células satélites quiescentes não expressam os MRFs, porém, sob a ação de estímulos como fatores de crescimento ou citocinas, ocorre a ativação desse tipo celular que prolifera e expressa os MRFs de maneira similar ao que ocorre com as células precursoras miogênicas durante a miogênese. Os mecanismos de crescimento muscular são regulados pela expressão temporal dos (MRFs), que controlam a expressão dos genes relacionados com o crescimento muscular.

Tipos de miosinas de linhagens de frangos de corte criados em sistemas de confinamento e semiconfinamento

O objetivo neste trabalho foi avaliar o peso vivo, o peso de pernas, os aspectos morfológicos das fibras musculares do músculo flexor longo do hálux e o perfil eletroforético das miosinas de cadeia pesada de quatro linhagens de frangos de corte criados nos sistemas de confinamento e semiconfinamento. Foram utilizados 1.440 pintos distribuídos em delineamento inteiramente casualisado em esquema fatorial 4 × 2, composto de quatro linhagens (Ross 305, Máster Gris, Label Rouge e Vermelhão Pesado) e dois sistemas de criação (confinamento e semiconfinamento), cada combinação avaliada com quatro repetições. Aos 28 e 84 dias de idade, foram abatidas quatro aves por tratamento, totalizando 64 aves. A eletroforese identificou a presença das três isoformas de miosinas, tipo MyHC-I, MyHC-IIa e MyHC-IIb, no músculo flexor longo do hálux dos frangos de corte. Com aumento da idade, a isoforma de miosina MyHC-II aumenta, enquanto a MyHC-I diminui. Somente aos 84 dias de idade, a expressão das isoformas de miosina do tipo MyHC-II foram influenciadas pela linhagem, confirmando o reflexo da seleção na linhagem Ross no músculo mais glicolítico.A linhagem Ross apresenta maior peso vivo, peso de perna, peso e área do músculo flexor longo do hálux em comparação às linhagens tipo caipira.

Gill chemoreceptors and cardio-respiratory reflexes in the neotropical teleost pacu, Piaractus mesopotamicus

This study examined the location and distribution of O-2 chemoreceptors involved in cardio-respiratory responses to hypoxia in the neotropical teleost, the pacu (Piaractus mesopotamicus). Intact fish and fish experiencing progressive gill denervation by selective transection of cranial nerves IX and X were exposed to gradual hypoxia and submitted to intrabuccal and intravenous injections of NaCN while their heart rate, ventilation rate and ventilation amplitude were measured. The chemoreceptors producing reflex bradycardia were confined to, but distributed along all gill arches, and were sensitive to O-2 levels in the water and the blood. Ventilatory responses to all stimuli, though modified, continued following gill denervation, however, indicating the presence of internally and externally oriented receptors along all gill arches and either in the pseudobranch or at extra-branchial sites. Chemoreceptors located on the first pair of gill arches and innervated by the glossopharyngeal nerve appeared to attenuate the cardiac and respiratory responses to hypoxia. The data indicate that the location and distribution of cardio-respiratory O-2 receptors are not identical to those in tambaqui (Colossoma macropomum) despite their similar habitats and close phylogenetic lineage, although the differences between the two species could reduce to nothing more than the presence or absence of the pseudobranch.

Occurrence of biflagellate spermatozoa in Cetopsidae, Aspredinidae, and Nematogenyidae (Teleostei : Ostariophysi : Siluriformes)

In the present study spermiogenesis was investigated in Cetopsis coecutiens (Cetopsidae), and Bunocephalus amazonicus (Aspredinidae), while spermatozoa ultrastructure was investigated in C. coecutiens, B. amazonicus, and Nematogenys inermis (Nematogenyidae). Aspredinidae and Cetopsidae share a spermatogenesis of the semicystic type, and a particular type of spermiogenesis process not reported in any fish group. In the three species analyzed, spermatozoa are biflagellate with flagella having the classical axoneme formulae (9 + 2). The analysis of thirteen characters showed the presence of eight characters shared by Cetopsidae and Aspredinidae, and six characters shared by Cetopsidae and Nematogenyidae, which may suggest that these three families may be more related than actually hypothesized, comprising a very primitive siluriform lineage originated after Diplomystidae.

Taxonomic status of the genus Sotalia: Species level ranking for tucuxi (Sotalia fluviatilis) and costero (Sotalia guianensis) dolphins

Dolphins of the genus Sotalia are found along the Caribbean and Atlantic coasts of Central and South America and in the Amazon River and most of its tributaries. At present, the taxonomy of these dolphins remains unresolved. Although five species were described in the late 1800s, only one species is recognized currently (Sotalia fluviatilis) with two ecotypes or subspecies, the coastal subspecies (Sotalia fluviatilis guianensis) and the riverine subspecies (Sotalia fluviatilis fluviatilis). Recent morphometric analyses, as well as mitochondrial DNA analysis, suggested recognition of each subspecies as separate species. Here we review the history of the classification of this genus and present new genetic evidence from ten nuclear and three mitochondrial genes supporting the elevation of each subspecies to the species level under the Genealogical/Lineage Concordance Species Concept and the criterion of irreversible divergence. We also review additional evidence for this taxonomic revision from previously published and unpublished genetic, morphological, and ecological studies. We propose the common name costero for the coastal species, Sotalia guianensis (Van Beneden 1864), and accept the previously proposed tucuxi dolphin, Sotalia fluviatilis (Gervais, 1853), for the riverine species.

Floral development and embryology in the early-divergent grass Pharus

A detailed study of floral ontogeny, anatomy, and embryology in two (of six) species of Pharus is presented as part of a series of comparative investigations on early-divergent grasses. Pharus is a taxonomically isolated genus belonging to the earliest-diverging grass lineage with a true grass spikelet. It is unusual in possessing remarkably dimorphic florets: male florets possess two lodicules, six stamens, and a pistillode, whereas female florets lack lodicules entirely but possess six staminodes and a tricarpellary ovary with three stigmas. The rudimentary lodicules in male florets are initiated after the stamen whorls. There are most commonly six androecial organs, but in some florets, a five-staminate condition was observed, resulting from suppression of the abaxial stamen from the inner whorl, or even a four-staminate condition resulting from subsequent fusion of the two adaxial outer stamens (i.e., elements of both whorls). Thus, the pattern of floral zygomorphy in Pharus differs from that of many other grasses. Centrifixed anther attachment is reported for the first time in Pharus, resembling the condition in another early-divergent grass, Anomochloa, though anthers are introrse in Anomochloa compared with latrorse in Pharus. Anther wall development is of the reduced type in Pharus, in contrast to most other monocots. Microsporogenesis is of the successive type, as in many other monocots. The ovary develops from three distinct primordia and is unilocular with a single ovule and a pronounced ovary beak that is highly characteristic of Pharus. There is a hollow style, in contrast to the solid styles that are common in many other grasses. The embryo is highly differentiated, as in other grasses, with a distinct epiblast and a small cleft between the scutellum and the coleorhiza.

Alterations of spermatogenesis in etoposide-treated rats: A stereological study

The etoposide is an anticancer drug that interacts with topoisomerase II. Thirty-day-old rats received intraperitonially 2mg/kg of etoposide for 30 consecutive days. Their testes were analyzed in the adult phase under light microscopy according to histomorphometric and stereological parameters. Random 3mum-thick-paraplast sections of testis were stained with periodic acid-Schiff reaction and Harris' hematoxylin method. Serum testosterone level and reproductive performance were also investigated. The results showed an accentuated decrease in the frequency of germinal lineage cell types and differentiated spermatogonia were the most affected cell types. Morphometric and stereological testicular parameters exhibited highly, significant reductions in adult etoposide-treated rats. Their reproductive performance diminished but their serum testosterone level was not significantly altered. The mortality frequency of the progenies was 100%.

Reassessment of the phylogenetic position of conulariids (?Ediacaran-Triassic) within the subphylum medusozoa (Phylum Cnidaria)

Fossil taxa of uncertain phytogenetic affinities can play a crucial role in the analysis of character evolution within major extant groups. Marques & Collins (2004) concluded that conulariids (?Ediacaran-Triassic) are an extinct group of medusozoan cnidarians most closely related to Stauromedusae. However, only six of the 87 characters used by these authors can be observed in conulariid fossils. Rescoring the character states of conulariids in a conservative manner yields a new hypothesis for the phylogenetic position of conulariids, namely that they are the sister group of the scyphozoan order Coronatae rather than Stauromedusae, which is revealed as the earliest diverging lineage of Medusozoa. This new hypothesis also implies several different sequences of character evolution within Cnidaria. Specifically, the presence of a periderm completely covering the polyp in conutariids and coronates appears to be derived within Scyphozoa. Strobilation appears to be a synapomorphy uniting conulariids, Coronatae, Rhizostomeae and Semaeostomeae. This result supports the controversial interpretation of one exceptionally preserved conulariid that potentially shows that these animals produced ephyrae by strobilation. Finally, the pelagic adult medusa stage and the giant fibre nerve net appear to be features that are derived within Medusozoa.

Root surface conditioning with nicotine or cotinine reduces viability and density of fibroblasts in vitro

The purpose of study was to evaluate fibroblast attachment and cellular morphology on root surfaces chemically conditioned with nicotine or cotinine. A secondary objective was to determine if mechanical scaling and root planning of these chemically conditioned surfaces would alter cellular attachment. Root surface dentin specimens were prepared from uniradicular teeth of non-smoking patients. Specimens were randomly assigned to two experimental groups: no treatment (chemical conditioning only) and scaling and root planning after conditioning (SRPC). The concentrations of the tested substances were in the range of 0-1 mg/mL (nicotine) and 0-1 ?g/mL (cotinine). After a 24-h conditioning period, dentin slices were incubated with continuous lineage of fibroblastic cells from rat (McCoy cells) for another 24 h. Specimens were prepared for SEM analysis and microphotographs. The statistical analysis of the data indicated significant alteration of cellular morphology on fibroblasts that were grown on root surface exposed to nicotine concentrations greater than 1 ? g/mL. This effect of nicotine was not reduced by SRPC. on the other hand, in the SRPC group cellular density was greater. For cotinine-conditioned specimens, the greater concentrations also led to alteration on morphology, and these alterations were observed in the SRPC group as well. Cotinine did not induce significant changes on cellular density. The results indicated that fibroblasts are negatively influenced by nicotine present on the dentin substrate and also that scaling may reduce these effects. Cotinine treatment on root surfaces may alter cell morphology and density but these effects were less severe than that promoted by nicotine, and were not affected by scaling.

Root instrumentation with an erbium : yttrium-aluminum-garnet laser: Effect on the morphology of fibroblasts

Objective: the purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet laser instrumentation of root surfaces on the morphology of fibroblasts from continuous lineage. Method and materials: Dentinal slices with 4 mm(2) of surface area were obtained from teeth extracted for severe periodontal involvement. Specimens were assigned to one of three treatment groups: group 1, application of the laser with an energy level of 250 mJ at 103 pulses per second; group 2, application of the laser with an energy level of 80 mJ at 166 pulses per second; and group 3, similar to group 2, but with concomitant water irrigation of the device. The specimens were incubated in multiwell plates containing cell culture media. After 24 hours, the specimens were submitted to routine preparation for scanning electron microscopy. Three independent and blind examiners used photomicrographs to evaluate the morphology of the fibroblasts: 0 = without cells; 1 = flat cells; 2 = round cells; and 3 = combination of round and flat cells. Results: Statistical analysis indicated that there were significant differences among treatment groups and that group 3 was significantly different from groups 1 and 2. Conclusion: There was no difference between groups 1 and 2 in the morphology of fibroblasts. Laser instrumentation with concomitant irrigation impaired the adhesion of fibroblasts to dentinal surfaces.