Liquid Storage of Geophagus brasiliensis semen in the Presence of Different Extenders.

BACKGROUND: In order to preserve the genetic diversity of cichlid fish in gene banks, it is necessary to use certain extenders to maintain the integrity of spermatozoa cells during cooling. OBJECTIVE: To evaluate the effects of different extenders on the quality parameters of cooled semen of Geophagus brasiliensis. MATERIALS AND METHODS: Semen samples were collected from seven adult fish and diluted with five extenders: Beltsville Thawing Solution (BTS™), Hanks' Balanced Salt Solution (HBSS), Tris-glucose, Ginsburg's Fish Ringers, and Phosphate buffered Saline. All parameters were evaluated in fresh semen samples and after cooling at 4°C at 0, 24, 48, and 96 h to evaluate cell viability (membrane integrity, DNA, and mitochondrial functionality) and motility rate and weather motility. RESULTS: The BTS and Tris-glucose resulted in the best outcomes (P<0.05) in terms of membrane integrity assessments (35.1% and 30.9%, respectively), DNA integrity (71.6%; 75.7%), mitochondrial function (26.9%; 28.0%) and motility rate (8.6%; 8.6%), respectively, for semen cooled to 4°C for 96 h. However, the 48-h period motility after cooling in BTS was superior to all other treatments. CONCLUSION: BTS and Tris-glucose can be considered as the best extenders for the cold storage of Geophagus brasiliensis spermatozoa.

Embryonic and larval development of Brycon amazonicus (SPIX & AGASSIZ, 1829).

The objective of this study was to describe the embryonic and larval development of Brycon amazonicus, featuring the main events up to 50 hours after fertilization (AF). The material was provided by the Aquaculture Training, Technology and Production Center, Presidente Figueiredo (AM). The characterization was based on stereomicroscopic examination of the morphology of eggs, embryos and larvae and comparison with the literature. Matrinxã eggs are free, transparent, and spherical, with a perivitelline space of 0.56 ± 0.3 mm. The successive divisions give rise to cells with 64 blastomeres during the first hour AF. The gastrula stage, beginning 02 h 40 min AF, was characterized by progressive regression cells and the formation of the embryonic axis, leading to differentiation of the head and tail 05 h 30 min AF. From 06 to 09 h AF the somites, notochord, otic and optic vesicles and otoliths were observed, in addition to heart rate and the release of the tail. The larvae hatched at 10 h 30 min AF (29.9 °C), with a total length of 3.56 ± 0.46 mm. Between 19 and 30 h AF, we observed 1) pigmentation and gut formation, 2) branchial arches, 3) pectoral fins, 4) a mouth opening and 5) teeth. Cannibalism was initiated earlier (34 h AF) which was associated with rapid yolk absorption (more than 90% until 50 h AF), signaling the need for an exogenous nutritional source. The environmental conditions (especially temperature) influenced the time course of some events throughout the embryonic and larval development, suggesting the need for further studies on this subject.

Embryonic and larval development of Brycon amazonicus (SPIX & AGASSIZ, 1829) / Desenvolvimento embrionário e larval de Brycon amazonicus (SPIX & AGASSIZ, 1829)

Abstract The objective of this study was to describe the embryonic and larval development of Brycon amazonicus, featuring the main events up to 50 hours after fertilization (AF). The material was provided by the Aquaculture Training, Technology and Production Center, Presidente Figueiredo (AM). The characterization was based on stereomicroscopic examination of the morphology of eggs, embryos and larvae and comparison with the literature. Matrinxã eggs are free, transparent, and spherical, with a perivitelline space of 0.56 ± 0.3 mm. The successive divisions give rise to cells with 64 blastomeres during the first hour AF. The gastrula stage, beginning 02 h 40 min AF, was characterized by progressive regression cells and the formation of the embryonic axis, leading to differentiation of the head and tail 05 h 30 min AF. From 06 to 09 h AF the somites, notochord, otic and optic vesicles and otoliths were observed, in addition to heart rate and the release of the tail. The larvae hatched at 10 h 30 min AF (29.9 °C), with a total length of 3.56 ± 0.46 mm. Between 19 and 30 h AF, we observed 1) pigmentation and gut formation, 2) branchial arches, 3) pectoral fins, 4) a mouth opening and 5) teeth. Cannibalism was initiated earlier (34 h AF) which was associated with rapid yolk absorption (more than 90% until 50 h AF), signaling the need for an exogenous nutritional source. The environmental conditions (especially temperature) influenced the time course of some events throughout the embryonic and larval development, suggesting the need for further studies on this subject.

Resumo O objetivo deste trabalho foi descrever o desenvolvimento embrionário e larval do Brycon amazonicus, caracterizando os principais eventos ocorridos até 50 h Após Fertilização (AF). O material é proveniente do Centro de Treinamento, Tecnologia e Produção em Aqüicultura em Presidente Figueiredo (AM). A caracterização foi feita com base na análise estereomicroscópica da morfologia dos ovos, embriões e larvas e comparação bibliográfica. Os ovos da matrinxã são livres, transparentes, esféricos com espaço perivitelínico de 0,56 ± 0,3 mm. As sucessivas clivagens originam células com 64 blastômeros na primeira hora AF. A gástrula, iniciada 02 h e 40 min AF caracterizou-se por progressiva involução celular e formação do eixo embrionário, culminando com diferenciação de cabeça e cauda com 05 h 30 min AF. De 06 às 09h AF foi observada a formação de somitos, notocorda, vesículas óptica, ótica e otólitos, além de batimentos cardíacos e liberação da cauda. As larvas eclodiram com 10 h 30 min AF (29,9 °C), com 3,56 ± 0,46 mm de comprimento total. Entre 19 e 30 h AF foram observadas: 1) pigmentação e formação do tubo digestivo 2) surgimento de arcos branquiais 3) nadadeira peitoral 4) abertura da boca e 5) surgimento dos dentes. O canibalismo iniciou mais precocemente (34 h AF), em relação aos trabalhos existentes com o gênero, o que associado à rápida absorção do vitelo (mais de 90% até 50 h AF), sinaliza a necessidade de ofertar recurso alimentar exógeno. As condições ambientais (especialmente temperatura e pH) influenciaram na abreviação de alguns eventos ao longo do desenvolvimento embrionário e larval, sugerindo a necessidade de estudos complementares a esse respeito.

Comportamento ingestivo de ovinos alimentados com diferentes fontes de fibra / Ingestive behavior of sheep fed with different sources of fiber

Avaliou-se o efeito de dietas distintas sobre o comportamento ingestivo (alimentação, ruminação e ócio) em ovinos. Dezesseis carneiros, castrados, confinados em gaiolas de metabolismo, com peso médio de 45,1kg, foram distribuídos aleatoriamente em quatro tratamentos: T1- silagem de cana + concentrado; T2- silagem de cana + concentrado + 15% de caroço de algodão; T3- silagem de cana e T4- feno de Tifton 85. Os animais foram submetidos à observação visual durante 24 horas a intervalos de 10 minutos. Houve diferença entre os tratamentos com relação aos tempos despendidos com alimentação e ócio, sendo que o tratamento com feno de Tifton 85 apresentou os mais altos valores, 6,04 h/dia, para o tempo de alimentação, e os mais baixos valores para o tempo de ócio, 7,50 h/dia.

The effects of different diets on the ingestive feeding behavior in wethers (feeding, ruminating and idle) were evaluated. Sixteen wethers with an average weight of 45.1kg were housed in metabolic cages and were randomly assigned to four treatments (T1- sugarcane silage + concentrate; T2- sugarcane silage + concentrate + 15% whole cottonseed; T3- sugarcane and T4- hay Tifton 85). The ingestive behavior was assessed by visual observation at 10 minute intervals for 24 hours. There were differences between treatments for time spent feeding and idle, and the treatment with Tifton 85 hay showed the highest values for feeding time, 6.04 h/day, and lower values for idle time, 7.50h/day.

Use of amides as cryoprotectants in extenders for frozen sperm of tambaqui, Colossoma macropomum.

Amides were tested as cryoprotectants in comparison with glycerol and DMSO (more traditional cryoprotectants) for recovery of Colossoma macropomum (tambaqui fish) sperm. Milt was extended in Beltsville Thawing Solution, then frozen with the addition of 2%, 5%, 8%, or 11% of: (1) dimethylacetamide (DMA), (2) dimethylformamide (DMF), (3) methylformamide (MF), or with 5% glycerol or 10% dimethylsulfoxide. Fertilization rates were greatest (P<0.001) with amides; 8% DMF (91.6±1.3%), 5% DMF (88.9±1.6%), and 8% MF (83.0±1.6%), which did not significantly differ among themselves, when compared with glycerol (51.6±2.4%) and DMSO (61.9±3.1%). The best hatching rates (P<0.001) also occurred for 5% or 8% DMF and 8% MF (79.1±3.1, 87.6±1.5, and 74.8±3.0, respectively) and were also similar (P>0.05). For such treatments, both fertilization and hatching rates were similar (P>0.05) to those with fresh sperm (91.7±1.4 and 87.4±1.4, respectively). The best sperm motility across extenders (at least 55.7%) was with 5%, 8%, and 11% DMF (P<0.001). Those same treatments, along with 11% MF, provided the longest (P<0.001) period of motility (at least 1 min). The greatest sperm integrity (more than 54%) was with 5% and 11% MF and with DMA and DMF at all tested concentrations (P<0.001). The greatest (P<0.001) sperm viability (at least 31%) was for 5%, 8%, and 11% DMA, and with 8% and 11% MF, and also for DMF at all tested concentrations. Sperm DNA integrity was best (more than 50%) for 2%, 5%, and 8% MF and for DMA and DMF at all concentrations (P<0.001), whereas 2% DMA, 11% MF, 11% DMF, and the three amides at both 5% and 8% yielded the highest mitochondrial functionality (at least 44%; P<0.001); thus, 8% MF and both 5% and 8% DMF were the cryoprotectants with the best postthaw quality for C. macropomum sperm.

Anti-inflammatory and anti-ulcerogenic properties of Stachytarpheta cayennensis (L.C. Rich) Vahl.

In the present work, the anti-inflammatory and gastroprotective properties of ethanolic extracts of Stachytarpheta cayennesis (L.C. Rich) Vahl (Verbenaceae) were assessed. Chromatographic analysis of the crude ethanolic extract, SC01, revealed high concentrations of the iridoid ipolamiide, whereas the SC02, the second ethanolic extract, presented the arylpropanoid verbacoside as a major constituent. The oral administration of SC01 (100 mg/kg) into Swiss mice failed to inhibit paw oedema and pleural exudation induced by carrageenan and zymosan, whereas SC02 (100 mg/kg, p.o.) inhibited oedema and protein extravasation in all instances. Both extracts inhibited total leukocyte accumulation into the pleural cavity 4 and 24h after the intrathoracic (i.t.) injection of carrageenan, due to the inhibition of neutrophil and mononuclear cell influx, whereas only SC02 was able to inhibit leukocyte mobilization induced by zymosan (100 microg/cavity, i.t.). SC02 inhibited LPS (250 ng/cavity)-induced total leukocyte, neutrophil and eosinophil accumulation in the pleural cavity, whereas SC01 selectively inhibited neutrophil influx. In addition, our data indicates that the extract SC02 presents an important anti-ulcerogenic activity, since it inhibited diclofenac-induced (100 mg/kg, p.o.) gastric ulcera. Overall, these data provide evidence for the anti-inflammatory and gastroprotective properties of Stachytarpheta cayennensis, supporting its use in folk medicine for such purposes.

Antinociceptive effect of Nidularium procerum: a Bromeliaceae from the Brazilian coastal rain forest.

Nidularium procerum, a common plant of the Brazilian flora, has not yet been studied for its pharmacological properties. We report here that extracts of N. procerum show both analgesic and anti-inflammatory properties. Oral (p.o.) or intraperitoneal (i.p.) administration of an aqueous crude extract from leaves of N. procerum (LAE) inhibited the writhing reaction induced by acetic acid (ED50 value = 0.2 mg/kg body weight, i.p.) in a dose-dependent manner. This analgesic property was confirmed in rats using two different models of bradykinin-induced hyperalgesia; there was 75% inhibition of pain in the modified Hargreaves assay, and 100% inhibition in the classical Hargreaves assay. This potent analgesic effect was not blocked by naloxone, nor was it observed in the hot plate model, indicating that the analgesic effect is not associated with the activation of opioid receptors in the central nervous system. By contrast, we found that LAE (0.02 microg/ml) selectively inhibited prostaglandin E2 production by cyclooxygenase (COX)-2, but not COX-1, which is a plausible mechanism for the analgesic effect. A crude methanol extract from the leaves also showed similar analgesic activity. An identical extract from the roots of N. procerum did not, however, block acetic acid-induced writhes, indicating that the analgesic compounds are concentrated in the leaves. Finally, we found that LAE inhibited an inflammatory reaction induced by lipopolysaccharide in the pleural cavity of mice.

The intraperitoneal Plasmodium berghei-Pasteur infection of Swiss mice is not a system that is able to detect the antiplasmodial activity in the Pothomorphe plant extracts that are used as antimalarials in Brazilian endemic areas.

The antimalarial activity of the hexane and methanol extracts derived from the Brazilian plants Pothomorphe peltata and Pothomorphe umbellata-whose leaves are popularly employed in medicinal folk remedies for the treatment of malaria-was assessed through in vivo tests with the Peters method. The extracts were delivered to Plasmodium berghei-infected mice via the oral or the subcutaneous route. A suppressive effect on the parasitemia seemed to be evident when data regarding the intraperitoneal injection of Pothomorphe umbellata extracts were analyzed. However, a definitive conclusion on an effective antimalarial activity is not possible, as two distinct-"standard" and "slow"-patterns of parasitemia occurring at similar frequencies in both treated and untreated intraperitoneally infected mice were observed. Nevertheless, the existence of two distinct profiles of parasitemia was not clear among the animals that were infected via the intravenous route. These data indicate the need for further studies on the biological features of the host/parasite interaction in the intraperitoneally P. berghei-infected Swiss mice system to standardize the model and to improve its usefulness in the screening of antimalarial compounds.