Testosterone secretion and pharmacological spermatozoal recovery in the cane toad (Bufo marinus)

The cane toad (Bufo marinus) was used as a model to study male anuran reproductive endocrinology and to develop a protocol for non-invasive sperm recovery. Circulating testosterone concentrations in 6-hourly samples did not vary significantly (P < 0.05) over a 24 h period although there was a tendency (P = 0.06) for testosterone to be elevated at 19:00 h relative to other times of the day, which may be related to the nocturnal activity pattern of this species. Testosterone secretion after intraperitoneal (IP) injection of either a GnRH agonist (5 mu g IP) or hCG (1000 IU) was also examined. While the GnRH agonist did not produce a significant increase above basal plasma testosterone (0.29, 95% C.I. of 0.05-1.10 ng/ml), injection of hCG resulted in an increase (P < 0.01) of plasma testosterone with peak concentrations at approximately 120 min (4.17, 95% C.I. of 2.69-7.44 ng/ml) after injection. Non-invasive pharmaceutical sperm recovery was attempted following IP injection of graded doses of GnRH agonist, hCG or FSH. Urine was collected at 3, 6 and 12 h after treatment to assess sperm quality and quantity. The optimal protocol for sperm recovery in cane toads was injection of either 1000 or 2000 IU hCG; there was no significant difference in the quality of the spermic urine samples obtained using either dose of hCG or with respect to collection time. The findings indicated that hCG can be used to assess testicular steroidogenic status and also to induce sperm recovery in the cane toad. The hCG protocols developed in this study will have application in studies on the reproductive biology of rare and endangered male anurans. (c) 2005 Elsevier B.V. All rights reserved.

Effects of sodium on potassium nutrition in three tropical root crop species

The potassium (K) nutrition and high K requirement of tropical root crops may be affected by their sodium (Na) status, as has been observed in a number of plant species. Solution culture was used to study the effects of K and Na supplies in tannia [Xanthosoma sagittifolium (L.) Schott.], sweetpotato [Ipomoea batatas (L.) Lam.] and taro [Colocasia esculenta (L.) Schott]. At low K supply, Na ameliorated symptoms of K deficiency and increased growth in tannia, and to a lesser extent in sweetpotato, but not in taro. None of the species responded to Na at adequate K supply. Differences in response to Na were attributed to differences in Na translocation to plant tops. At maximum Na supply, the Na concentration in index leaves averaged 1.82% in tannia, 0.205% in sweetpotato, and 0.0067% in taro. An increase in the supply of Na resulted in a shift in the critical K concentration for deficiency (i.e., 90% of maximum yield) in index leaves from 2.9% to 1.2% in tannia, and from 4.8% to 2.5% in sweetpotato. The critical K concentration in taro was 3.3%, irrespective of Na supply. To overcome the problem in tannia and sweetpotato of determining the critical concentration relevant to a leaf sample of unknown K status, a relationship was established for each species relating the critical K concentration to the concentration of Na in the index leaves.

Environmental and biological drivers of feeding and spatial dynamics in the green sea urchin, Strongylocentrotus droebachiensis

In eastern Canada, the destruction of foundational kelp beds by dense aggregations (fronts) of the omnivorous green sea urchin, Strongylocentrotus droebachiensis, is a key determinant of the structure and dynamics of shallow reef communities. Current knowledge about factors affecting the ability of S. droebachiensis to exert top-down community control is based largely on observational studies of patterns in natural habitats, yielding fragmentary, and sometimes contradictory, results. The present research incorporated laboratory microcosm experiments and surveys of urchins in natural habitats to test the effects of abiotic (wave action, water temperature) and biotic (body size, population density) factors on: (1) individual and aggregative feeding on the winged kelp, Alaria esculenta; and (2) displacement, microhabitat use, distribution, and aggregation in food-depleted habitats. Wave action, water temperature, and body size strongly affected the ability of urchins to consume kelp: individual feeding increased with increasing body size and temperature, while aggregative feeding decreased with increasing wave action. Yet, feeding in large urchins dropped by two orders of magnitude between 12 and 18°C. Increasing wave action triggered shifts in urchin displacement, microhabitat use, distribution, and aggregation: urchins reduced displacement and abandoned flat surfaces in favour of crevices. They increasingly formed two-dimensional aggregations at densities ≥110 individuals m⁻². Collectively, results provide a foundational understanding of some of the drivers of feeding and spatial dynamics of S. droebachiensis and potential impacts on the formation of grazing fronts.

Fresh biomass of macroalgae collected at Hansneset, Kongsfjorden, Spitsbergen in 1996/1998

We surveyed macroalgae at Hansneset, Blomstrand in Kongsfjorden, Svalbard, down to 30 m depth between 1996 and 1998. In total, 62 species were identified: 16 Chlorophyta, 25 Phaeophyceae, and 21 Rhodophyta. The majority of species (53.5%) belonged to the Arctic cold-temperate group, followed in frequency by species distributed from the Arctic to the warm-temperate region (25.9%). Four endemic Arctic species (Laminaria solidungula, Acrosiphonia flagellata, A. incurva, and Urospora elongata) were found. Two species (Pogotrichum filiforme and Mikrosyphar polysiphoniae) were new to Svalbard. Chlorophyta, Phaeophyceae, and Rhodophyta extended from the eulittoral zone down to 11, 21, and >30 m depths with maximum biomasses at 1-5 m, 5-10 m, and 5-30 m depths, respectively. Annual and pseudoperennial species had highest biomasses in the upper 5 m, while perennials were distributed deeper. The highest biomass (8600 g/m**2 wet weight) at 5 m depth comprised mainly L. digitata, Saccorhiza dermatodea, Alaria esculenta, and Saccharina latissima. The biogeographic composition of macroalgae at Hansneset was rather similar to that of northeastern Greenland, but different from that of northern Norway, which has a higher proportion of temperate species. Climate warming and ship traffic may extend some of the distribution ranges of macroalgae from mainland Norway to Svalbard.

Kunitzins: Prototypes of a new class of protease inhibitor from the skin secretions of European and Asian frogs

Amphibian skin secretions contain biologically-active compounds, such as anti-microbial peptides and trypsin inhibitors, which are used by biomedical researchers as a source of potential novel drug leads or pharmacological agents. Here, we report the application of a recently developed technique within our laboratory to “shotgun” clone the cDNAs encoding two novel but structurally-related peptides from the lyophilized skin secretions of one species of European frog, Rana esculenta and one species of Chinese frog, Odorrana schmackeri. Bioanalysis of the peptides established the structure of a 17-mer with an N-terminal Ala (A) residue and a C-terminal Cys (C) residue with a single disulphide bridge between Cys 12 and 17, which is a canonical Kunitz-type protease inhibitor motif (-CKAAFC-). Due to the presence of this structural attribute, these peptides were named kunitzin-RE (AAKIILNPKFRCKAAFC) and kunitzin-OS (AVNIPFKVHLRCKAAFC). Synthetic replicates of these two novel peptides were found to display a potent inhibitory activity against Escherichia coli but were ineffective at inhibiting the growth of Staphylococcus aureus and Candida albicans at concentrations up to 160 μM, and both showed little haemolytic activity at concentrations up to 120 μM. Subsequently, kunitzin-RE and kunitzin-OS were found to be a potent inhibitor of trypsin with a Ki of 5.56 μM and 7.56 μM that represent prototypes of a novel class of highly-attenuated amphibian skin protease inhibitor. Substitution of Lys-13, the predicted residue occupying the P1 position within the inhibitory loop, with Phe (F) resulted in decrease in trypsin inhibitor effectiveness and antimicrobial activity against Esherichia coli, but exhibits a potential inhibition activity against chymotrypsin.

A importância dos ácidos fenólicos na atividade anti-inflamatória de cogumelos comestíveis

Os cogumelos comestíveis são uma fonte rica de moléculas bioativas que lhes conferem importantes atividades biológicas. Moléculas como os Polissacáridos, terpenóides e os compostos fenólicos têm sido descritos como os componentes mais importantes no que respeita á atividade anti-inflamatória dos cogumelos (1). No presente trabalho, os extratos etanólicos de cogumelos comestíveis foram obtidos por maceração e caracterizados quimicamente em termos de ácidos fenólicos por técnicas de HPLC-DAD. Além disso, derivados metilados e glucuronados dos ácidos fenólicos identificados foram também sintetizados com o objetivo de mimetizar reações de metabolização no organismo e estudar a capacidade destas moléculas de manter a bioatividade exibida inicialmente. Os extratos obtidos, os ácidos fenólicos e compostos sintetizados foram avaliados pela sua atividade anti-inflamatória. De entre as amostras analisadas, B. impolitus revelou o mais elevado conteúdo em ácidos fenólicos (675 ± 23 μg/g), seguido de C. cibarius > A. caesaria > L. deliciosus > B. aereus > M. esculenta > B. edulis; devido à contribuição do ácido cinâmico que foi encontrada em maior quantidade nesta amostra (505 ± 12 μg/g). Mais ainda, B. impolitus apresentou também maior inibição da produção de NO (EC50=166 ± 10 μg/mL) seguido das amostras A. caesaria > C. cibarius > L. deliciosus > M. esculenta > B. aereus > B. edulis. No que respeita aos compostos individuais, o ácido cinâmico (CA) revelou a atividade mais forte (EC50 = 182 ± 16 μM), seguido pelos ácidos p-hidroxibenzóico (HA) (239 ± 29 μM) e p-Cumárico (CoA) (442± 33 μM), o que realça a importância destas moléculas para a atividade anti-inflamatória dos cogumelos. Comparando a atividade exibida pelos ácidos fenólicos com os respetivos derivados, é possível verificar a seguinte ordem de atividades: ácido p-hidroxibenzóico: HA > HA-M3 > HA-M2 > HA-M1 > HA-G; ácido p-cumárico : CoA-M1 > CoA-G > CoA-M2 > CoA-M3 > CoA e ácido cinâmico: CA-G > CA > CA-M1. Perante os resultados obtidos é de realçar a importância dos ácidos fenólicos na contribuição para a bioatividade exibida pelos cogumelos em estudo. Mais ainda, foi possível concluir que as alterações das moléculas pelas reações de conjugação no organismo têm influência na bioativade das moléculas iniciais, sendo que muitas vezes esta atividade é aumentada.

Plantas forrageiras das caatingas: usos e potencialidades.

Coleta de analises; descricao e usos das especies: camaratuba, carqueija, catingueira-verdadeira, espinheiro/jacurutu/jiquiri, facheiro, favela/faveleira, feijao-brabo, imbuzeiro/umbuzeiro, juazeiro, jurema-preta, jurema-vermelha, macambira, mandacaru/mandacaru-de-boi, manicoba, marmeleiro/marmeleiro-preto, moleque-duro, mororo/unha-de-vaca, quebra-faca, sabia, sete-cascas/cascudo/pau-de-casca. Comentarios gerais e recomendacoes.