Assessing genetic diversity in cultured aquatic species : the Sydney Rock Oyster (Saccostrea glomerata) stock improvement program as a model

Genetic variation is the resource animal breeders exploit in stock improvement programs. Both the process of selection and husbandry practices employed in aquaculture will erode genetic variation levels overtime, hence the critical resource can be lost and this may compromise future genetic gains in breeding programs. The amount of genetic variation in five lines of Sydney Rock Oyster (SRO) that had been selected for QX (Queensland unknown) disease resistance were examined and compared with that in a wild reference population using seven specific SRO microsatellite loci. The five selected lines had significantly lower levels of genetic diversity than did the wild reference population with allelic diversity declining approximately 80%, but impacts on heterozygosity per locus were less severe. Significant deficiencies in heterozygotes were detected at six of the seven loci in both mass selected lines and the wild reference population. Against this trend however, a significant excess of heterozygotes was recorded at three loci Sgo9, Sgo14 and Sgo21 in three QX disease resistant lines (#2, #5 and #13). All populations were significantly genetic differentiated from each other based on pairwise FST values. A neighbour joining tree based on DA genetic distances showed a clear separation between all culture and wild populations. Results of this study show clearly, that the impacts of the stock improvement program for SRO has significantly eroded natural levels of genetic variation in the culture lines. This could compromise long-term genetic gains and affect sustainability of the SRO breeding program over the long-term.

Telomere dynamics in the Sydney rock oyster (Saccostrea glomerata): an investigation into the effects of age, tissue type, location and time of sampling.

Telomere length has been purported as a biomarker for age and could offer a non-lethal method for determining the age of wild-caught individuals. Molluscs, including oysters and abalone, are the basis of important fisheries globally and have been problematic to accurately age. To determine whether telomere length could provide an alternative means of ageing molluscs, we evaluated the relationship between telomere length and age using the commercially important Sydney rock oyster (Saccostrea glomerata). Telomere lengths were estimated from tissues of known age individuals from different age classes, locations and at different sampling times. Telomere length tended to decrease with age only in young oysters less than 18 months old, but no decrease was observed in older oysters aged 2-4 years. Regional and temporal differences in telomere attrition rates were also observed. The relationship between telomere length and age was weak, however, with individuals of identical age varying significantly in their telomere length making it an imprecise age biomarker in oysters.

PRELIMINARY STUDIES ON CRYOPRESERVATION OF SYDNEY ROCK OYSTER (SACCOSTREA GLOMERATA) LARVAE

In this study several parameters critical to the success of cryopreserving Sydney rock oyster (Saccostrea glomerata) larvae were investigated. They were: (1) cryoprotectants (10% dimethyl sulfoxide and 10% propylene glycol). (2) freezing protocols (with or without the seeding step). (3) larval concentrations (1,000, 3,000, 5,000, 10,000, 30,000 individuals mL(-1)). and (4) larval ages (6, 12, 24, 48 and 96 h old). The survival rates were determined as percentages of postthaw larvae performing active movements for the 6 and 12 h larvae or active cilia movement for the 24, 48 and 96 h larvae. Analyses showed that the difference in survival rates between different age classses was significant in all the experiments conducted, with the maximum survival rate being achieved in the 24-h-old larvae the postthaw survival rates of larvae cryopreserved with 10% dimethyl sulfoxide (93.1 +/- 0.2%) were significantly higher (P < 0.001) that those with 10% propylene glycol (81.5 +/- 0.4%). Differences in postthaw survival rates between different concentrations (1,000 30,000 individuals mL(-1)) were not significant within each of the three larval age classes (6-, 12-, and 24-h-old ) used.

Colletotrichum graminicola (Ces.) Wilson sobre pasto ovillo (Dactylis Glomerata L.) en la Argentina

p.127-130

Involvement of nitric oxide in the gastroprotective effects of an aqueous extract of Pfaffia glomerata (Spreng) Pedersen, Amaranthaceae, in rats

The plants belonging to Pfaffia genus are used in folk medicine to treat gastric disturbances. This study examined the effects of an aqueous extract of Pfaffia glomerata (Spreng) Pedersen (AEP) on the gastrointestinal tract. Wistar rats were pretreated orally (p.o.) with the AEP (125, 250, 500 and 1000 mg.kg(-1)) before induction of ulcers by hypothermic restraint stress (HRS, 3 h restraint stress at 4 degreesC), ethanol (ET, 70%; 0.5 ml/animal; p.o.) or indomethacin (IND, 20 mg.kg(-1); s.c.). Control animals received water (C) or ranitidine (60 mg.kg(-1)) p.o. The AEP protected rats against HRS and ET-induced ulcers, but was not able to protect the gastric mucosa against IND-induced ulcers. When injected into the duodenal lumen, the AEP reduced total acidity and both basal and histamine-stimulated acid secretion in pylorus-ligated rats. In addition, gastric secretion from AEP-treated animals exhibited increased concentrations of nitrite and nitrate. Treatment of animals with L-NAME (120 mg.kg(-1), p.o.) prevented both the reduction of total acidity and the increase in NO, levels promoted by AEP treatment. In conclusion, AEP effectively protected the gastric mucosa and inhibited gastric acid secretion in rats, probably by involving the histaminergic pathway and an enhanced production of nitric oxide in the stomach. (C) 2003 Elsevier B.V. All rights reserved.

Produção de β-ecdisona em Pfaffia glomerata (Spreng.) Pedersen em função da adubação orgânica em 6 épocas de crescimento

O trabalho teve como objetivo verificar a influência da adubação orgânica em 6 épocas de crescimento na produção de β-ecdisona por plantas de Pfaffia glomerata. O experimento foi conduzido na Fazenda Santo Antonio do Araquá, distrito de Catâneo Ângelo, município de São Manuel, São Paulo, Brasil. Utilizou-se o delineamento de blocos ao acaso, num esquema fatorial 5x6, com quatro repetições, considerando-se 8 plantas úteis por parcela. Os blocos foram constituídos de 6 épocas de crescimento (60, 120, 180, 240, 300 e 360 dias após a germinação) e de 5 doses de esterco de galinha curtido [testemunha (sem adubação), 15, 30, 45 e 60 t ha-1]. Após cada colheita, as raízes das plantas foram secas em estufa com circulação de ar forçada a 40ºC e pesadas para posterior extração do β-ecdisona, seguindo metodologia desenvolvida por Magalhães (2000). Os resultados foram submetidos à análise de variância e ao teste de separação de médias de Scott Knott, todos a 5% de probabilidade. Quando ocorreu interação os resultados foram avaliados usando-se análise de regressão polinominal. O teor de β-ecdisona não foi influenciado pelas doses de adubo e nem pela época do crescimento das plantas. Porém a quantidade total de β-ecdisona por raiz foi influenciada pela época de crescimento, sendo que aos 360 dias após a emergência ocorreu uma maior quantidade do princípio ativo em todos os tratamentos. Apesar de não diferir estatisticamente dos demais tratamentos, aos 360 dias após a emergência das plantas, o tratamento 30 t ha-1 foi o que proporcionou maior quantidade de β-ecdisona.

Influência de diferentes meios de cultura sobre o crescimento de Pfaffia glomerata (Spreng.) Pedersen (Amaranthaceae) para conservação in vitro

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

Antidiarrhoeal effects of Mikania glomerata Spreng. (Asteraceae) leaf extract in mice

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of Mikania glomerata (Asteraceae) leaf extract combined with anti-venom serum on experimental Crotalus durissus (Squamata: Viperidae) envenomation in rats

Crotalic envenomation represents the highest number of deaths when compared to other snakebite envenomations of medical interest. Crotalic venom has important characteristics such as neurotoxicity, myotoxicity, nephrotoxicity, and clotting and hemolytic action. We evaluated the clinical and laboratory aspects of Crotalus durissus terrificus experimental envenomation in Wistar rats treated with antivenom and the aqueous extract of the plant mikania glomerata. The animals were divided into three groups: Group C (control); Group VS-venom and antivenom; Group VSM-venom, antivenom and aqueous extract of M glomerata. Crotalic poison caused clinical and laboratory alterations in Wistar mice. Significant clinical alterations were: temperature decrease, edema in the venom inoculated member, sedation and a locomotion decrease in groups VS and VSM when compared with group C. A faster recovery from sedation was observed only for animals of group VSM when compared to VS. There was an increase in the number of leukocytes, neutrophils and creatine kinase in the VS and VSM groups, compared to group C. Wistar rats showed a high resistance to crotalic venom. Additional studies with different doses, time of treatment, different administration methods and histopathological and immunological studies are necessary to understand the action of M glomerata in crotalic accidents. Rev. Biol. Trop. 57 (4): 929-937. Epub 2009 December 01.

Photoacclimation in a tropical population of Cladophora glomerata (L.) Kützing 1843 (Chlorophyta) from southeastern Brazil

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

Collection of Fafia [Pfaffia glomerata (Spreng.) Pedersen] in northwestern state of Parana-Brazil

Interviews were conducted with fafia (Pfaffia glomerata (Spreng.) Pedersen) collectors from the municipal districts of Querencia do Norte and Porto Rico, where enterprises and buyers of this plant are concentrated. The results allowed us to conclude that both the adults and children make collection. Each adult collects from 50 to 150 kg of roots/day, depending on the collection area, for about 8 months/year. Collections mostly occur all over the year, but the activity becomes more intense from May to August. All families are not exclusively dedicated to fafia collection and also develop other rural activities. They have been collecting fafia for 2 to 13 years, indicating that an intensive exploitation has been present in the region for over a decade. During collection, no plant part is used for replanting the species. The roots are commercialised by regional buyers. The price of the roots varies from US $ 0.07 to US $ 0.13/kg and average gain is about US $ 2,055/family/year, representing a considerable profit for the collectors.

Influence of fertilizer on coumarin content and biomass production in Mikania glomerata sprengel

Mikania glomerata Sprengel leaf extracts have been used in cough syrup formulations due to the presence of active coumarin derivatives. Yields of such natural compounds in cultivated plants can be affected by several nutritional factors. To determine the effect of fertilization on biomass productivity, organ proportion and coumarin content, Mikania glomerata plants were submitted during one year to treatments with organic (humus or manure) and inorganic (different levels of nitrogen) fertilizers. Coumarin concentration was raised by organic fertilization, inorganic nutrients induced increased phytomass (stem and leaf) yield, however no fertilized plants showed enhanced leaf production.

Vegetative propagation of Mikania glomerata: Micro-propagation and cuttings

The scope of this work was to compare two systems for vegetative propagation: conventional one (from cut stems) and in vitro micropropagation from axillary buds. Nodal segments (1 cm) of Mikania glomerata were used as explants. The experiments were evaluated in relation to number of shoots; % of rooting; number of roots and total fresh weight. Multiple shoots developed in MS containing 0.5 mg/L BAP. Rooting was induced in the presence of 1.0 mg/L IBA. Stems with five buds and one pair of leaves were the most appropriate for the production of cuttings. The time necessary for developing a protocol for the production of M. glomerata micropropagated plantlets was 6 months, whereas only half time was required to produce plantlets from stem cuttings. The greatest problem met during micropropagation was the culture contamination by endophytic bacteria and fungi.

Coumarin production in Mikania glomerata callus

Cell cultures of Mikania glomerata Sprengel were established with leaf segments cultured on White medium supplemented with 1 mg/L BA and 3 mg/L NAA. Different types and concentrations of growth regulators were tested for callus maintenance. Determination of coumarin content was performed in HPLC using authentic coumarin standard. Growth regulator concentration affected biomass and coumarin accumulation. Cultures developed in semisolid medium containing both BA and NAA exhibited enhanced biomass production as well as coumarin accumulation. In the most favorable conditions tested, cells accumulated 25 μg/g of dry weight what is much inferior to the yield already reported in intact plants (5 mg/g of dry weight). However, results obtained so far suggest several alternatives for culture manipulation in order to optimize the productivity of coumarin by M. glomerata cultured cells.

Seasonal variation in coumarin content of Mikania glomerata

Mikania glomerata Spengal extract contains approximately 0.5 percent of coumarin, a substance that displays anti-inflammatory and expectoratory activities. Extracts from different young leaves of plants collected during the early evening of December and July contained the highest levels coumarin. Plants dried in oven with circulating air contained 7.3 mg of coumarin/g of dry weight, demonstrating that this was the most appropriate drying condition for Mikania glomerata.