Development and characterization of a new set of 164 polymorphic EST‐SSR markers for diversity and breeding studies in rubber tree (Hevea brasiliensis Müll. Arg.)

Despite its economic importance and recent genome release, the need for molecular tools for Hevea brasiliensis is high. In the frame of a disease resistance study, EST sequences were retrieved from public database or generated by sequencing SSH libraries. Sequences were trimmed and microsatellite motifs searched using an ad hoc bioinformatic pipeline, and pairs of primers for the amplification of candidate markers were generated. We found a total of 10 499 unigenes from both sources of sequences, and 673 microsatellites motifs were detected using the default parameters of the pipeline. Two hundred sixty‐four primer pairs were tested and 226 (85.6%) successfully amplified. Out of the amplified candidate markers, 164 exhibited polymorphism. Relationships based on dendrograms using simple matching index and diversity statistics based on EST‐SSRs were compared with Genomic SSRs, showing the potentialities of EST‐derived microsatellites for resistance studies but also for population genetics approaches.     

Identification of Volatiles of the Dinoflagellate Prorocentrum cordatum

The dinoflagellate Prorocentrum cordatum, often called P. minimum, is a potentially toxic alga found in algal blooms. Volatile compounds released by the alga might carry important information, e.g., on its physiological state, and may act as chemical messengers. We report here the identification of volatile organic compounds emitted by two strains, xenic P. cordatum CCMP 1529 and axenic P. cordatum CCMP 1329. The volatiles released during culture were identified despite their low production rates, using sensitive methods such as open-system-stripping analysis (OSSA) on Tenax TA desorption tubes, thermodesorption, cryofocusing and GC/MS-analysis. The analyses revealed 16 compounds released from the xenic strain and 52 compounds from the axenic strain. The majority of compounds were apocarotenoids, aromatic compounds and small oxylipins, but new natural products such as 3,7-dimethyl-4-octanolide were also identified and synthesized. The large difference of compound composition between xenic and axenic algae will be discussed.     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.     

Antibody response in goats experimentally infected with Toxoplasma gondii

Serum samples of five goats inoculated with Toxoplasma gondii were analyzed using the enzyme linked immunosorbent assay (ELISA), indirect hemagglutination (IHA) and western blotting (WB). Antibodies detected by ELISA peaked between 19 and 62 days after inoculation and persisted throughout the experiment with no association to parasitaemia. Using WB, the main antigens detected had molecular weights of approximately 68, 62, 50, 48, 42, 34, 28, 26, 22 and 19 kDa. Antibody titers of between 1:256 and 1:32000 were observed using IHA, with a significant drop in activity after treatment with 2-mercapto-ethanol between days 12 and 48. This coincided with the parasitaemic period that occurs between 5 and 64 days after inoculation.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

温度对大叶桃花心木(Swietenia macrophylla King)幼苗叶片的光合影响

本文研究了大叶桃花心木（Swietenia macrophylla King）一年生幼苗在经过夜温处理后的光响应曲线和在饱和光强下的CO2反应曲线.结果表明:在大气CO2浓度下,叶片的最佳光合作用温度在25-31℃之间,而在饱和CO2浓度下为31-35℃.在25℃以下光合速率开始降低,主要是由于羧化效率的降低,而当温度超过31℃时,光合速率下降,是因为羧化效率的降低和呼吸速率的增加.CO2浓度对光合的促进作用在低温下受到抑制,这意味着未来在CO2浓度增高的情况下,高浓度的CO2对热带常绿植物光合的促进在冬天低温情况下表现不十分明显.图4参23.     

First feeding of diploid and triploid yellowtail tetra Astyanax altiparanae: An initial stage for application in laboratory studies

In this study, the aim was to establish a protocol for first feeding of diploid and triploid yellowtail tetra Astyanax altiparanae in laboratory conditions. The fry were fed with five different diets: (i) Artemia franciscana nauplii, (ii) plankton, (iii) dry food, (iv) Artemia franciscana nauplii + plankton, and (v) Artemia nauplii + plankton + dry food. Additionally, the growth and survival rates of diploid and triploid individuals were also evaluated. On day 10, the length of the fish between the treatments differed significantly (p = .0001) and ranged from 4.07 ± 0.06 mm (dry food) to 8.50 ± 0.64 mm (plankton + Artemia). The sizes of the fish increased with time, except for the fish fed with dry food. The survival rates were similar for the fish fed with the four diets and ranged from 80.7 ± 5.4% (dry food + plankton + Artemia to 92.0 ± 1.6% (plankton + Artemia), but differed from the fish fed with dry food (17.7 ± 5.8%, p = .0017). Diploids and triploids did not present differences on day 0 (p = .2252) and on day 10 (p = .4844) when the fish presented 6.77 ± 0.25 mm and 6.54 ± 0.15 mm respectively. Survival of diploids (87.3 ± 5.13%) and triploids (74.67 ± 2.30%) were also similar (p = .0285). These data are innovative and useful for establishing protocols for this species in both academic and applied sciences.