Temperature-sex determination in Podocnemis expansa (Testudines, Podocnemididae)

This study has been carried out at the central region of the Araguaia river on the border between the states of Goiás and Mato Grosso in the Brazilian Amazon Basin from September to December 2000. We recorded temperature fluctuation, clutch-size, incubation period and hatching success rate and hatchlings' sex ratio of five nests of Podocnemis expansa (Schweigger, 1812). Despite the relatively small sample size we infer that: a) nests of P. expansa in the central Araguaia river have a lower incubation temperature than nests located further south; however, incubation period is shorter, hatching success rate is lower and clutch-size is larger; b) Podocnemis expansa may present a female-male-female (FMF) pattern of temperature sex-determination (TSD); c) thermosensitive period of sex determination apparently occur at the last third of the incubation period; and, d) future studies should prioritize the relationship between temperature variation (i.e., range and cycle) and embryos development, survivorship and sex determination.

Anfíbios Anuros do Distrito Federal

The frogs of the Federal District of Brazil are listed and discussed as to habit, biology and ecology. The F. D is situated 22° 54' 24" S. & 43° 10' 21" W Gr. and comprises 1.356 km². Its topography includes sea-shore, maritime scrub, lagoons, plains and marsh, open slopes, forested mountains and great heads of rock. Three thousand feet of altitude are attained at two points. Fifty two different frogs occur in the F.D. Three fifths of them live in open country. Two fifths of these have never been found above the plains; the others range higher but mostly in open country. Their environment offers conditions suitable for average tadpoles and adults. these frogs are more or less unspecialized. There are six genera and thirty species. Two thirds of the latter belong to the type genera of the large neotropical families Bufonidae, Leptodactylidae and Hylidae. Only in the maritime scrub formation are conditions somewhat different. Water for average tadpoles is provided by the lagoons. The xerophytism of the vegetation is, however, so marked that bromeliads growing on the ground provide almost the only appropriate shelter for adult tree-frogs used to sleeping upright on the vegetation. One large Hylid genus lives entirely in them. It is casque-headed and phragmotic, shutting the lumen of the leaf-cup with head used as a plug. Another large Hylid genus shows a lesser degree of the same specialization. (Lutz A & Lutz B, 1939 II). One genus with two species is entirely saxicolous; it lives on wet ledges of rock at all phases of its life history. (B. Lutz 1948). The other two fifths of the frogs from F. D. are montane forest forms. Their environment offers numerous and varied biotopes and is near optimum for adults. There is,however, hardly any standing water available for larvae. These frogs are ecologically diversified. They also show a general trend towards spawning in the adult biotipe, which leads to delayed hatching, semi-aquatic and terrestrial larvae and direct development. (B Lutz, 1948). The author interprets the morphological specialization of the casque-headed Hylids and the biological specialization of the montane forest forms as adaptive. Casque-headedness and phragmosis increase protection against blood-suckers and predators. The humidity of the rain forest permits eggs, embryos and larvae to develop, unharmed, outside their usual, aquatic, environment.

Effects of Laurus nobilis (Lauraceae) on Biomphalaria glabrata (Say, 1818)

Experiments were carried out using aqueous extracts from leaves and flowers of Laurus nobilis on Biomphalaria glabrata. Treatments were performed on blastula stage (± 15 h after first cleavage) and on adult snails (11-18 mm). In both instances they were exposed for 24 h to different concentrations of the extracts on snails (200 to 2500 ppm) and embryos (20 to 300 ppm) at 25 ± 1ºC. The embryos were observed for a period of 20 days after treatment and the snails for 10 days. Results obtained with leaf aqueous extracts have shown a degree of toxicity on embryos starting at a concentration of 125 ppm, the flower extract being effective at 35 ppm. The malformation obtained with the different concentrations falls into the unespecific type category, however some cephalic and shell malformations were found in embryos treated with concentrations over 50 ppm (leaves) and 25 ppm (flowers). The LD90 on adult snails obtained by treatments with flower and leaf extract was observed at concentrations of 340 ppm and 1900 ppm respectively.

Establishment and Characterization of a Cell Line from the Mosquito Anopheles albimanus (Diptera: Culicidae)

A new cell line designated LSB-AA695BB, was established from embryos of the mosquito Anopheles albimanus. The primary culture was initiated in April, 1995, and the first passage was made 48 days later. Serial subcultures of the cells have been carried through 90 passages from Abril 1995 to February 1996. The cells were grown at 28°C in MK/VP12 medium, supplemented with 20% fetal bovine serum; the pH tolerance ranged between 6.8 to 7.0. The cells have also been adapted to MM/VP12 medium under the same pH, temperature and serum concentration. The majority of the cells were a fibroblast-type. Isozyme characterization showed a pattern similar to that of An. albimanus pupae and adults but distinct from Ae. taeniorhynchus and Ae. albopictus (C6/36) mosquito cell lines. The culture was shown to be free of mycoplasma, bacteria and fungi. Microsporidia contamination of transovarial transmission was controlled with 6.0 mg/ml of albendazole

The Influence of Self-fertilization performance and Copulation Behaviour in Reproduction by Cross-fertilization in Groups of Biomphalaria tenagophila (Mollusca, Planorbidae)

The following hypotheses were tested for groups of simultaneous hermaphrodites Biomphalaria tenagophila: (a) snails that have low reproductive success during the process of self-fertilization do not increase their reproductive success after the end of grouping; (b) the copulation behaviour and the presence of one snail whose eggs have a low viability rate influence the partner's reproductive success by cross-fertilization. Groups were constituted by a homozygous pigmented snail and two albinos: one with a viability rate higher than 70% ("good reproducers") and the other less than 10% ("bad reproducers"). All pigmented snails had viability rates higher than 70%. The "good" and "bad" reproducer albino snails had similar copulation behaviour. However, after the end of grouping, the "bad reproducers" continued to have viability rates less than 10% over 30 days. In 100% of the cases that pigmented snails copulated (performing either a male role or simultaneously male and female roles) exclusively with "good" reproducer albinos, they presented high reproductive success (producing, on average of 8.4 pigmented embryos/egg-mass). However, in 100% of the cases that pigmented snails copulated with both partners, the "good" reproducer albino snails produced none or very few embryos (the highest average was 2.2 pigmented embryos/egg-mass). Therefore, the production of viable embryos by cross-fertilization was more influenced by self-fertilization performance than by copulation behaviour. The presence of a snail whose eggs have a low viability rate could decrease their partners reproductive success

Temperature influence on embryonic development of Anopheles albitarsis and Anopheles aquasalis

Temperature influence on the embryonic development of Anopheles aquasalis and An. albitarsis was investigated. At 26ºC, 75% and 60% of respectively An. aquasalis and An. albitarsis eggs hatched, with one peak of eclosion, between the 2nd and 3rd day after oviposition. At 20 ± 2ºC, around 66-70% of An. aquasalis eggs hatched, with one eclosion peak, on the 5th day. On the other hand, An. albitarsis eclosion at 21 ± 2ºC decreased to 10-22%, with two eclosion peaks, on the 4th-5th day and on the 9th-12th day. These data indicate a stronger temperature influence over An.albitarsis than over An. aquasalis embryos.

Consumo e custo da alimentação para recuperação da atividade ovariana luteal cíclica de vacas mestiças leiteiras com anestro

O objetivo do trabalho foi verificar o consumo e o custo de alimentos para recuperação da atividade ovariana luteal cíclica (AOLC) em vacas mestiças Holandês x Zebu com anestro. Foram usadas 18 vacas, não-gestantes, não-lactantes, magras, apresentando ovários sem função luteal, de tamanho normal e sem folículos palpáveis na superfície. Doze animais permaneceram em confinamento e receberam alimentação para ganho de peso até o reinício da AOLC. Os seis animais restantes, constituindo o grupo controle, receberam alimentação de mantença para o baixo peso apresentado e permaneceram em anestro durante o período experimental. A AOLC foi avaliada pela concentração de progesterona no soro sangüíneo (coleta de sangue a cada sete dias), pelo exame dos ovários por palpação retal a cada 12 dias, e observação visual dos sinais do estro três vezes ao dia. A recuperação da AOLC nas vacas com anestro exigiu um consumo médio de 722,4 kg de matéria seca, 50,4 kg de proteína bruta e 402,4 kg de NDT, relativos à ingestão média de 2.374,2 kg de volumoso e 194,1 kg de concentrado. O custo desses alimentos somado à perda estimada da produção de leite numa vaca de 3.000 litros de leite/lactação, provocada pelo prolongamento do intervalo de partos, foi equivalente a 1.364,2 litros de leite (preço recebido pelo produtor = R$ 0,20/litro). Esse custo elevado da recuperação do anestro onera o custo final da produção de leite, tendo em vista a alta incidência de anestro nos rebanhos brasileiros.

Peso para recuperação da atividade ovariana luteal cíclica em vacas leiteiras mestiças em anestro

Procurou-se conhecer o ganho de peso necessário e o peso mínimo para restabelecimento da atividade ovariana luteal cíclica (AOLC) em vacas adultas, mestiças Holandês x Zebu, não-lactantes, magras e com ovários inativos, anestro adquirido após longo período de restrição alimentar. Seis animais, com peso médio de 322,0 ± 27,0 kg, receberam dieta de mantença para o baixo peso apresentado (grupo I) e 12 animais, com peso médio de 315,0 ± 29,4 kg, foram alimentados para ganho de peso até a recuperação da AOLC (grupo II). O sangue foi coletado (dosagem de progesterona-RIA), e os animais, pesados semanalmente. A AOLC foi avaliada pela concentração de progesterona no sangue, exame dos ovários por palpação retal a cada 12 dias e observação visual do estro três vezes ao dia. O reinício da AOLC ocorreu nos animais do grupo II quando pesaram em média 392,7 ± 29,4 kg. A média do ganho de peso total nesse grupo foi de 77,7 ± 11,2 kg, correspondendo a 24,7 ± 4,5% do peso desses animais em anestro, ou 37,7% dos 206,2 kg perdidos na fase de restrição alimentar para adquirir anestro. Os seis animais do grupo I permaneceram em anestro. Os resultados mostram a influência do nível alimentar sobre a função luteal ovariana e a necessidade de ganho de peso para o restabelecimento do ciclo estral em vacas mestiças leiteiras, magras e em anestro.

Efeito do pastejo restringido em aveia sobre a produção de leite

O objetivo deste trabalho foi avaliar o efeito do pastejo restringido em aveia (Avena sp.) sobre a produção de leite. Foram utilizadas 12 vacas mestiças holandês-zebu, distribuídas em blocos ao acaso, em dois tratamentos: A. pastejo restringido em aveia (três horas dia-1) mais 10 kg de silagem de milho; B. silagem de milho à vontade como único volumoso. Todos os animais receberam 5,0 kg de concentrado cabeça-1 dia-1 com 19% de proteína bruta (PB) e 75% de nutrientes digestíveis totais (NDT). Não houve diferença (P > 0,05) entre os tratamentos A e B na produção média de leite (14,6 vs 13,3 kg vaca-1 dia-1). Houve diferença (P < 0,05) no teor de gordura (3,4 vs. 3,8%), porém o mesmo não afetou (P > 0,05) a produção de leite corrigida para 4% de gordura. Houve diferença (P < 0,08 ) no ganho de peso vivo. Os animais que pastejaram aveia ganharam 0,53 kg dia-1, e as vacas que consumiram silagem de milho como único volumoso ganharam 0,25 kg dia-1. Concluiu-se que o pastejo restringido na cultivar de aveia São Carlos permite elevada produção de leite com vacas mestiças.

Plant regeneration from protoplast of Brazilian citrus cultivars

A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 muM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 muM), NAA (1.07 muM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development.

Daily intake of lactating crossbred cows grazing elephant grass rotationally

The goal of this trial was to estimate the total dry matter (TDMI) and daily pasture dry matter intakes (PDMI) by lactating crossbred Holstein - Zebu cows grazing elephant grass (Pennisetum purpureum Schum.) paddocks submitted to different rest periods. Three groups of 24 cows were used during two years. The paddocks were grazed during three days at the stocking rate of 4.5 cows/ha. Treatments consisted of resting periods of 30 days without concentrate and resting periods of 30, 37.5 and 45 days with 2 kg/cow/day of 20.6% crude protein concentrate. From July to October, pasture was supplemented with chopped sugarcane plus 1% urea. Total daily dry matter intake was estimated using the extrusa in vitro dry matter digestibility and the fecal output with chromium oxide. Regardless of the treatment the estimated average TDMI was 2.7, 2.9 and 2.9±0.03% and the mean PDMI was 1.9, 2.1 and 2.1±0.03% of body weight in the first, second and third grazing day, respectively (P<0.05). Only during the summer pasture quality was the same whichever the grazing day. Sugarcane effectively replaced grazing pasture, mainly in the first day when pasture dry matter intake was lowest.

Efeito de concentração espermática e período de incubação oócitoespermatozóides na fecundação in vitro em bovinos da raça Gir

Avaliou-se o efeito de concentrações espermáticas e períodos de incubação na fecundação in vitro com sêmen de touros Gir (Bos indicus). Oócitos maturados in vitro foram fecundados com sêmen de dois touros, nas concentrações de 1, 2 ou 4x10(6) espermatozóides/mL, por 12 ou 18 horas, e avaliada a taxa de fecundação e polispermia. Posteriormente, foi avaliada a taxa de clivagem com 72 horas e de blastocisto no oitavo dia pósfecundação. Um segundo experimento verificou o efeito de 1, 2 e 4x10(6) espermatozóides/mL por 18 horas, utilizando sêmen de três touros separadamente, bem como o efeito de 12 e 18 horas de incubação, na concentração de 1 a 2x10(6) espermatozóides/mL. Observou-se maior (P<0,05) polispermia na concentração de 4x10(6) espermatozóides/mL e que a associação dessa concentração com 18 horas aumentou a taxa de clivagem (P<0,05). No segundo experimento, o aumento do período de incubação melhorou (P<0,05) a taxa de clivagem para um dos touros e, quando comparados entre si, os animais produziram taxa de clivagem e de blastocistos diferentes (P<0,05) em uma mesma concentração ou período de incubação. Os resultados revelaram que a polispermia é o principal efeito do aumento da concentração espermática e que a eficiência entre touros na fecundação in vitro é dependente da concentração e período de incubação.

Morfogênese e acúmulo de biomassa foliar em pastagem de capim-elefante avaliada em diferentes épocas do ano

O objetivo deste trabalho foi avaliar o efeito da época do ano nas características morfogênicas e estruturais e no acúmulo de biomassa foliar de uma pastagem de capim-elefante (Pennisetum purpureum Schum.). Vacas mestiças Holandês x Zebu foram manejadas segundo o método de pastejo com lotação rotacionada com três dias de ocupação e 30 dias de descanso. Foi usado delineamento em blocos casualizados com seis repetições. As avaliações foram realizadas em fevereiro/março, abril/maio, julho/agosto e outubro de 2001. As maiores taxas de alongamento e aparecimento de folhas ocorreram durante fevereiro/março. Os perfilhos aéreos superaram os basilares em quantidade, mas apresentaram menores taxas de alongamento (5,1 versus 9,8 cm/dia/perfilho), aparecimento (0,13 versus 0,16 folhas/dia/perfilho) e senescência (0,9 versus 1,3 cm/dia/perfilho) foliares. As produções e taxas de acúmulo de biomassa foliar foram maiores durante fevereiro/março e apresentaram estreita relação com as variáveis morfogênicas. Os perfilhos aéreos contribuíram, em média, com 63% da biomassa foliar do capim-elefante.

Association of bGH and Pit-1 gene variants with milk production traits in dairy Gyr bulls

The objective of this study was to obtain genetic marker information in the Gyr breed by analyzing bGH and Pit-1 gene polymorphisms and to verify their association with milk production traits. One sample including 40 Gyr bulls were genotyped at two bGH gene restriction sites (bGH- AluI and bGH-MspI) and at one restriction site in the Pit-1 gene (Pit-1 HinfI). The bGH-MspI(-) allele was favorable for fat milk percentage. The heterozigous Pit-1 HinfI (+/-) bulls were superior for fat milk production, in relation to homozigous Pit-1 HinfI (+/+). The Pit-1 and bGH genes are strong candidates in the dairy cattle QTL search, and zebuine populations are promising samples for this purpose.

Polyembryony and identification of Volkamerian lemon zygotic and nucellar seedlings using RAPD

The objectives of this work were to evaluate the frequency of polyembryony, and to identify zygotic and nucellar seedlings of Citrus volkameriana using RAPD. Twenty-five polyembryonic and eight monoembryonic seeds were cultivated in vitrofor six months. DNA from seedlings was extracted and used in combination with five RAPD primers to identify zygotic or nucellar origin of the seedlings. Environmental conditions of the year affected significantly (P<0.05) the morphological characteristics of fruitsand the number ofembryos per seed. Polyembryonic seeds ranged from 30.9%, 44.8% to 54.4% over three years. Morphological characteristic was not correlated with polyembryony. In vitro culture enable all embryos of each seed to grow, favoring the percentage of seedlings identified as zygotic. In polyembryonic and monoembryonic seeds, 25.9% and 87.5% of the seedlings, respectively, were sexually originated. In polyembryonic seeds, not all zygotic seedlings were produced by small embryos located at the micropyle.