RESUMO O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.

ABSTRACT The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.

RESUMO O objetivo deste trabalho foi avaliar o efeito do rolipram durante a maturação de oócitos bovinos, expressão gênica e embriões produzidos in vitro. Os ovários bovinos foram coletados no matadouro. Os COCs foram selecionados e divididos em cinco grupos: controle 0 tempo; controle: MIV por 24 horas; tratamentos rolipram com bloqueio MIV por 24 horas em meio de maturação contendo 100, 150 e 200µM. Após 24 horas, todos os grupos foram recolocados em MIV por mais 24 horas. Subsequentemente COCs foram submetidos ao mesmo sistema MIV e fertilizados, sendo avaliada a taxa de clivagem e de blastocisto, além da expressão dos seguintes genes: Mater, BMP15 e Bax. Nenhuma diferença foi observada na expressão gênica. Dos oócitos avaliados logo após a aspiração folicular, 79,0% estavam em GV, GVBD, MI, enquanto 13,40% estavam em MII, e 7,60% em D/NI. A diferença significativa foi observada em diferentes concentrações (T100, T200 e T150µM) em oócitos que atingiram a fase MII em comparação aos tratamentos de controle (P=0,3). Diferenças foram observadas nas taxas de clivagem (P<0,5) entre T150 e T200 quando comparadas com as taxas do grupo C/24. Uma grande diferença foi observada na taxa de blastocisto (P<0,1) entre os tratamentos em relação ao grupo controle.

ABSTRACT The aim of this work was to evaluate the effect of the Rolipram during the maturation of bovine oocytes and gene expression of embryos produced in vitro. Bovine ovaries were collected in slaughterhouse. The COCs were selected and divided into 5 groups: Control 0 time; Control: IVM for 24 hours; Rolipram treatments with IVM blocking for 24 hours in maturation medium containing (100, 150 and 200µM). After 24 hours all groups were reseated in IVM for another 24 hours. Subsequently COCs were subjected to the same IVM system and fertilized, being checked for cleavage post fertilization and for blastocyst. In addition, performed expression of the following genes: Mater, BMP15 and Bax. No difference was found in gene expression. Of oocytes evaluated shortly after follicular aspiration, 79.00% were in GV, GVBD, MI, while 13.40%, were in MII and 7.60%, D/NI. Significant difference was observed in different concentrations (T100, T200 and T150µM) in oocytes that have reached the MII phase compared to control treatments (P= 0.003). Differences were observed in cleavage rate (P< 0.05) between T150 and T200 when compared to the C/24 Group. A high difference was observed on blastocyst rate (P< 0.001) among treatments compared to the control group.

Resumen En el estudio se evaluó el establecimiento aséptico de Monstera acuminata Koch y Monstera deliciosa Liebm (Araceae) a partir de hojas y la inducción de la organogénesis in vitro de M. acuminata K. a partir de discos de tallos de brotes jóvenes. Para el efecto se aplicaron diferentes protocolos de desinfección a hojas maduras y brotes jóvenes, de los cuales se extrajeron explantes foliares de 1 cm2 y discos de tallo de 1 mm de grosor, aproximadamente. Los explantes se establecieron en medios semisólidos con diferentes tratamientos hormonales durante la etapa de establecimiento aséptico e inducción de la organogénesis. La desinfección con hipoclorito de sodio (NaClO) al 3% por 20 min y el medio Murashige y Skoog (1962) al 50% con el preservador para cultivo de tejidos vegetales (PPM) favoreció una menor oxidación en los explantes foliares de ambas especies. Todos los explantes de M. deliciosa en ambos tratamientos cultivados en medio adicionado con PPM y a diferentes protocolos de desinfección sobrevivieron, no presentaron contaminación y más de 80% conservó actividad celular hasta 49 días de edad de cultivo. A los 35 días de cultivo, con la desinfección en Tween-20 + Etanol al 20% + NaClO al 2.5%, y la siembra de los explantes en el medio MS adicionado con 1 mg/l de BAP, 0.5 mg/l de AIA y 0.1 mg/l de ANA se indujeron siete brotes nuevos de discos de tallo. Monstera deliciosa presentó mayor capacidad de adaptación a condiciones in vitro. Los avances en el establecimiento aséptico y en la inducción de la organogénesis en aráceas nativas para la producción de mimbre, son la base para la conservación ex situ de las poblaciones locales.

Abstract In this paper we present the bases for the aseptic establishment of Monstera acuminata Koch and Monstera deliciosa Liebm (Araceae) from leaves and the induction of in vitro organogenesis of M. acuminata K. from discs of stems of young shoots. Different disinfection protocols were applied to mature leaves and young shoots, from which foliar explants of 1 cm2 of both species were extracted, and stem disks approximately 1 mm of thick in M. acuminata K. The explants were established in semi-solid media with different hormonal treatments during the stage of aseptic establishment and induction of organogenesis. Disinfection with 3 % of sodium hypochlorite (NaClO) for 20 min and 50 % Murashige y Skoog medium (1962) with PPM, led to less oxidation in the leaf explants of both species. All the explants in the two treatments belonging to M. deliciosa, grown in medium added with PPM and different disinfection protocols, survived, did not present contamination and more than 80 % maintained answer’s capacity until 49 days of culture. After 35 days of culture, with disinfection in Tween 20 + Ethanol 20% + 2.5% NaClO, and sowing of the explants in the MS medium added with 1 mg/l of BAP, 0.5 mg/l of AIA and 0.1 mg/l of ANA were indueces seven new shoots of stem disks. M. deliciosa had better ability to adapt to in vitro conditions. Advances in the establishment and induction of organogenesis in native araceas for wicker production are the basis for ex situ conservation of local populations.

Abstract The Tacaná-Boquerón region (TBR) in Chiapas is considered an area of high biodiversity in Mexico, with a rich but poorly studied orchid flora, which is an important component of the vegetation and threatened by an accelerated rate of deforestation. By means of fieldwork, and the revision of scientific collections and literature, an orchid checklist for the TBR and adjacent areas was made; using geographic information systems the distribution of this orchid flora was analyzed in the study area. From 1,235 records we report 105 genera and 325 species, two of them determined to infraspecific level and two are natural hybrids; nine species were additions to the Mexican flora; 39 species are considered at risk in Mexico; the national distribution of a further 20 species are restricted to the TBR. This orchid flora represents 24 % and 44 % of the national and state orchid species richness, respectively, is the second richest in Mexico, and only surpassed by the region El Momón-Las Margaritas-Montebello (Chiapas). The greatest orchid richness is concentrated between elevations of 500 to 2,499 m, in areas now given over to permanent agriculture (including coffee and cocoa plantations); in primary forest the richness is lower. The Tacaná Volcano Biosphere Reserve is the only protected area in the TBR and the proposal to extend it to the Boquerón peak would promote the protection of the orchid flora growing above 1,000 m in the region. Traditional coffee plantations could be alternative to conserve orchids that grow below 1,600 m.

Resumen La Región Tacaná-Boquerón (RTB) en Chiapas se considera un área de alta biodiversidad en México, posee una orquideoflora rica pero poco estudiada aunque es un componente importante de su vegetación, la cual está amenazada por un acelerado proceso de deforestación. Mediante trabajo de campo y revisiones de colecciones científicas y literatura se elaboró un listado de las orquídeas de la RTB y áreas adyacentes. Usando sistemas de información geográfica se analizó la distribución de esta orquideoflora en la zona de estudio. A partir de 1,235 registros se reportan 105 géneros y 325 especies, dos de ellas determinadas a nivel infraespecífico y dos híbridos naturales; nueve especies resultaron ser adiciones a la flora mexicana; 39 orquídeas son especies consideradas en riesgo en México, otras 20 restringen su distribución nacional a la RTB. Esta orquideoflora representa 24 % y 44 % de la riqueza orquideológica nacional y estatal, respectivamente, es la segunda más rica en México, solo superada por El Momón-Las Margaritas-Montebello (Chiapas). La mayor riqueza de orquídeas se concentra entre 500 y 2,499 m de elevación, en zonas actualmente destinadas a la agricultura permanente (incluyendo plantaciones de café y cacao); en los bosques primarios la riqueza es menor. La Reserva de la Biosfera del Volcán Tacaná es la única área protegida en la RTB, la propuesta de extenderla hasta el cerro Boquerón favorecería la protección de la orquideoflora que crece arriba de 1,000 m en la región; los cafetales tradicionales podrían ser alternativas para conservar orquídeas que crecen por debajo de 1,600 m.

Halophilic microorganisms are able to grow in the presence of salt and are also excellent source of enzymes and biotechnological products, such as exopolysaccharides (EPSs) and polyhydroxyalkanoates (PHAs). Salt-tolerant bacteria were screened in the Organic Composting Production Unit (OCPU) of São Paulo Zoological Park Foundation, which processes 4 ton/day of organic residues including plant matter from the Atlantic Rain Forest, animal manure and carcasses and mud from water treatment. Among the screened microorganisms, eight halotolerant bacteria grew at NaCl concentrations up to 4 M. These cultures were classified based on phylogenetic characteristics and comparative partial 16S rRNA gene sequence analysis as belonging to the genera Staphylococcus, Bacillus and Brevibacterium. The results of this study describe the ability of these halotolerant bacteria to produce some classes of hydrolases, namely, lipases, proteases, amylases and cellulases, and biopolymers. The strain characterized as of Brevibacterium avium presented cellulase and amylase activities up to 4 M NaCl and also produced EPSs and PHAs. These results indicate the biotechnological potential of certain microorganisms recovered from the composting process, including halotolerant species, which have the ability to produce enzymes and biopolymers, offering new perspectives for environmental and industrial applications.

A enxertia em porta-enxertos resistentes é uma técnica frequentemente recomendada para a cultura do pepino em áreas infestadas com nematoides-das-galhas. O presente trabalho teve como objetivo avaliar a produção de pepino enxertado em dois porta-enxertos comparado com pé-franco em solo infestado com Meloidogyne incognita raça 2 ou com M. javanica. Foram avaliados nove tratamentos (fatorial 3x3: pepino híbrido Tsuyataro pé-franco, enxertado na abóbora híbrida 'Shelper' e enxertado na abóbora híbrida 'Excitte Ikki' x sem inoculação, inoculação com M. incognita e inoculação com M. javanica), no delineamento em blocos ao acaso, com quatro repetições e seis plantas por parcela. Para as características de produção de frutos (número de frutos totais e comerciais por planta, massa de frutos totais e comerciais por planta) não foi obtida interação entre os fatores. Obteve-se maior produção de frutos em plantas enxertadas sobre a abóbora 'Shelper', independente da espécie de nematoide, enquanto que a enxertia sobre 'Excitte Ikki' não diferiu do pé-franco para produção total. Ambas as espécies de nematoide reduziram a produção de frutos, porém este efeito foi mais pronunciado com M. javanica. Tanto o pepino pé-franco como as plantas enxertadas foram suscetíveis, pois permitiram a multiplicação dos nematoides, com fator de reprodução, em avaliação realizada aos 72 dias após a inoculação, variando de 3,57 a 15,04, sendo que para M. javanica este valor foi maior no pepino pé-franco.

Grafting on resistant rootstock is a technique often recommended for cucumber crop in root-knot nematodes infested areas. We studied the yield of grafted cucumber on two rootstocks compared with non grafted in soil infested with Meloidogyne javanica or with M. incognita race 2. Nine treatments (factorial 3x3: Japanese cucumber hybrid Tsuyataro without grafting, grafted on squash hybrid Shelper and grafted on squash hybrid Excitte Ikki x without nematode inoculation, inoculated with M. incognita and inoculated with M. javanica) were evaluated in a randomized block design, with four replications and six plants per plot. There was no interaction between factors for yield evaluated characteristics (total and marketable fruit number per plant, total and marketable fruit weight per plant). Higher fruit yield was obtained in cucumber grafted on 'Shelper', for both nematode species, while cucumber grafted on 'Excitte Ikki' did not differ from non grafted plants for total fruit weight. Both nematode species reduced fruit yield, which was more evident when M. javanica was inoculated. Grafted and non grafted plants were susceptible, because they enabled the multiplication of nematodes, with a reproduction factor, at 72 days after inoculation, ranging from 3.57 to 15.04, with the highest value in cucumber non grafted inoculated with M. javanica.

In order to define the phytotoxic potential of Salvia species a database was developed for fast and efficient data collection in screening studies of the inhibitory activity of Salvia exudates on the germination of Papaver rhoeas L. and Avena sativa L.. The structure of the database is associated with the use of algorithms for calculating the usual germination indices reported in the literature, plus the newly defined indices (Weighted Average Damage, Differential Weighted Average Damage, Germination Weighted Average Velocity) and other variables usually recorded in experiments of phytotoxicity (LC50, LC90). Furthermore, other algorithms were designed to calculate the one-way ANOVA followed by Duncan's multiple range test to highlight automatically significant differences between the species. The database model was designed in order to be suitable also for the development of further analysis based on the artificial neural network approach, using Self-Organising Maps (SOM).

SUMMARY: Considering the importance of the diagonal artery for myocardial vascularization and the existence of a series of similar features between the hearts of men and pigs, this reserch was carried out to make a morphometric study of the diagonal artery in hearts of pigs. For this study 100 hearts of pigs were used. Coronary arteries of these hearts were injected with 15% gelatin solution stained with red chess. Next, the hearts were fixed in 10% formol solution for 10 days and dissectd to allow visualization of the diagonal artery and measurement of the ventricular height. It was considered diagonal artery only the vessel emerging from the angle between the anterior interventricular and circunflex arteries. It was observed that the diagonal artery in the hearts of pigs was present in 20% of the samples. Its lenght varied from 11,33 mm to 39,5 mm, with mean of 22,4 mm, being the majority (95%) described as artery of short course on the subepidardial plane. As for its behaviour, it was noted that it gave rise to a single branch in 50% of the cases, two branches in 45% of the cases and in one case (5%) it gave rise to three brances. It can be concluded that in case of absence of the diagonal artery the circunflex artery gives rise to many branches, possibly to play the role of irrigation suplly of the left ventricle.

RESUMEN: Debido a la importancia de la arteria diagonal en la vascularización del miocardio y a la existencia de características semejantes entre el corazón del hombre y del porcino, realizamos un estudio morfométrico de la arteria diagonal en 100 corazones de porcinos. Se inyectaron las arterias coronarias con una solución de gelatina al 15%, teñida de rojo. Luego, los corazones fueron fijados en solución de formol al 10%, por un período de 10 días y disecados para visualización de la arteria diagonal y comparación de la altura ventricular. Sólo se consideró como arteria diagonal al vaso que emergía de la bisectriz del ángulo formado por la arteria interventricular anterior y circunfleja. La arteria diagonal estuvo presente en el 20% de los casos. Su longitud varió entre 11,3 mm y 39,5 mm (x 22,4 mm), por lo que se clasificó a la mayoría de ellas (95%), como arterias de corto trayecto en el plano subepicárdico. Con relación a su ramas, se verificó que dicha arteria emitía una rama en el 50% de los casos, 2 ramas en el 45% y 3 ramas en el 5%. Se puede concluir entonces, que en el caso de ausencia de la arteria diagonal, la arteria circunfleja emitiría varias ramas, seguramente para suplir la insuficiente irrigación del ventrículo izquierdo.

Realizou-se inquérito sorológico e epidemiológico para cisticercose em indivíduos de cinco municípios da região Norte do Estado do Paraná, Brasil. De 2.180 indivíduos investigados através da reação de imunofluorescência indireta, 69 (3,2%) apresentaram títulos significativos de anticorpos anti-Cysticercus cellulosae. Os percentuais de indivíduos com títulos significativos encontrados em Sarandi (6,6%) e Marialva (4,7%) não diferem estatisticamente (Z=1.319, P=0,0936), mas diferem dos percentuais encontrados em Mandaguaçu, Paiçandu e Maringá (P<0,01). Destes indivíduos, 47,9% estavam na faixa etária de 21 a 49 anos e 79,4% eram do sexo feminino. Foi comum o relato de queixas como "dores de cabeça" (70,6%), "tonturas" (57,4%) e "convulsões" (7,4%), além de história de teníase (22,1%) e hábitos de ingestão de carne crua bovina (41,2%) ou suína (27,9%) e carne com "canjiquinha" (25,0%).

An epidemiological and serological study was carried out on a sample of 2,180 individuals, in five counties in the north of Paraná State - Brazil, using the indirect immunofluorescence test to detect anti-Cysticercus cellulosae antibodies. These individuals, 69 (3.2%) showed significant titers of antibodies. No single significant difference between the proportion of reactivity in Sarandi (6.6%) and in Marialva (4.7%) was observed (Z=1,319, P=0,0936), but it was significantly higher than that observed in Mandaguaçu, Paiçandu and Maringá (P<0.01). Of these individuals, 47.9% were within 21-49 years old and 79.4% were of female sex. "Headache" (70.6%), "faintness" (57.4%), and "convulsions" (7.4%) were among the most frequent by reported, moreover, cases of Taenia infections (22.1%) and the custom of eating uncooked beef (41.2%) or pork (27.9%) and meat containing cysticerci (25.0%) were also related.

Realizou-se inquérito sorológico e epidemiológico para cisticercose em indivíduos de cinco municípios da região Norte do Estado do Paraná, Brasil. De 2.180 indivíduos investigados através da reação de imunofluorescência indireta, 69 (3,2%) apresentaram títulos significativos de anticorpos anti-Cysticercus cellulosae. Os percentuais de indivíduos com títulos significativos encontrados em Sarandi (6,6%) e Marialva (4,7%) não diferem estatisticamente (Z=1.319, P=0,0936), mas diferem dos percentuais encontrados em Mandaguaçu, Paiçandu e Maringá (P<0,01). Destes indivíduos, 47,9% estavam na faixa etária de 21 a 49 anos e 79,4% eram do sexo feminino. Foi comum o relato de queixas como "dores de cabeça" (70,6%), "tonturas" (57,4%) e "convulsões" (7,4%), além de história de teníase (22,1%) e hábitos de ingestão de carne crua bovina (41,2%) ou suína (27,9%) e carne com "canjiquinha" (25,0%).

An epidemiological and serological study was carried out on a sample of 2,180 individuals, in five counties in the north of Paraná State - Brazil, using the indirect immunofluorescence test to detect anti-Cysticercus cellulosae antibodies. These individuals, 69 (3.2%) showed significant titers of antibodies. No single significant difference between the proportion of reactivity in Sarandi (6.6%) and in Marialva (4.7%) was observed (Z=1,319, P=0,0936), but it was significantly higher than that observed in Mandaguaçu, Paiçandu and Maringá (P<0.01). Of these individuals, 47.9% were within 21-49 years old and 79.4% were of female sex. "Headache" (70.6%), "faintness" (57.4%), and "convulsions" (7.4%) were among the most frequent by reported, moreover, cases of Taenia infections (22.1%) and the custom of eating uncooked beef (41.2%) or pork (27.9%) and meat containing cysticerci (25.0%) were also related.