Molecular characterization of canine coronavirus strains circulating in Brazil

To characterize canine coronavirus (CCoV) circulating in diarrheic puppies in Brazil, 250 fecal samples collected between 2006 and 2012 were tested. By using RT-PCR to partially amplify the M gene, CCoV RNA was detected in 30 samples. Sequence analysis of the M protein grouped eight strains with CCoV-I and another 19 with CCoV-II prototypes. To genotype/subtype the CCoV strains and assess the occurrence of single or multiple CCoV infections, RT-PCR of the S gene was performed, and 25/30 CCoV-positive strains amplified with one or two primer pairs. For 17/25 samples, single infections were detected as follows: six CCoV-I, nine CCoV-IIa and two CCoV-IIb. Eight samples were positive for more than one genotype/subtype as follows: seven CCoV-I/IIa and one CCoV-I/IIb. Sequence analysis revealed that the CCoV-I and IIa strains shared high genetic similarity to each other and to the prototypes. The Brazilian strains of CCoV-IIb displayed an aminoacid insertion that was also described in CCoV-IIb-UCD-1 and TGEV strains. Among the 25 CCoV-positive puppies, five had a fatal outcome, all but one of which were cases of mixed infection. The current study is the first reported molecular characterization of CCoV-I, IIa and IIb strains in Brazil.     

Evaluation of different approaches to individual tree growth and survival modelling using data collected at irregular intervals - a case study for Pinus patula in Kenya

Background： The minimum set of sub-models for simulating stand dynamics on an individual-tree basis consists of tree-level models for diameter increment and survival. Ingrowth model is a necessary third component in uneven-aged management. The development of this type of model set needs data from permanent plots, in which all trees have been numbered and measured at regular intervals for diameter and survival. New trees passing the ingrowth limit should also be numbered and measured. Unfortunately, few datasets meet all these requirements. The trees may not have numbers or the length of the measurement interval varies. Ingrowth trees may not have been measured, or the number tags may have disappeared causing errors in tree identification. Methods： This article discussed and demonstrated the use of an optimization-based approach to individual-tree growth modelling, which makes it possible to utilize data sets having one or several of the above deficiencies. The idea is to estimate all parameters of the sub-models of a growth simulator simultaneously in such a way that, when simulation begins from the diameter distribution at the first measurement occasion, it yields a similar ending diameter distribution as measured in the second measurement occasion. The method was applied to Pinus patula permanent sample plot data from Kenya. In this dataset, trees were correctly numbered and identified but measurement interval varied from 1 to 13 years. Two simple regression approaches were used and compared to the optimization-based model recovery approach. Results： The optimization-based approach resulted in far more accurate simulations of stand basal area and number of surviving trees than the equations fitted through regression analysis. Conclusions： The optimization-based modelling approach can be recommended for growth modelling when the modelling data have been collected at irregular measurement intervals.     

Insights into population genetics,connectivity and demographic history of the longnosed skate Dipturus oxyrinchus (Linnaeus, 1758) in the western Mediterranean Sea

1. Genetic information is crucial for the conservation of Dipturus oxyrinchus (Linnaeus, 1758), a threatened large skate with declining populations over most of its geographical range. The main aim of the present study was to investigate the genetic structure, connectivity and demographic history of the longnosed skate in Sardinia (western Mediterranean Sea).
2. Patterns of population structure were assessed in 175 specimens from six sampling sites. Variation in two mitochondrial genes (cytochrome c oxidase subunit I (COI) and control region) highlighted high genetic diversity and low but significant genetic differentiation among sites, which clustered into three groups corresponding to the north‐west, north‐east and south Sardinian coasts.
3. The observed genetic structuring could presumably depend on a combination of past geological events, contemporary restrictions to dispersal and biological characteristics of the species (e.g. site‐fidelity, no pelagic larval stage, limited dispersal of juveniles and/or adults).
4. Demographic analyses showed signs of past population expansion, but substantial current stability of Sardinian populations. From a conservation perspective, these results are encouraging, and indicate that Sardinian populations are still large and stable, and seem not to have suffered negative side‐effects from the ever‐growing fishing pressure in the region.
5. The occurrence of genetic structuring strongly supported the close monitoring of populations to identify any erosion of their gene pool, and high genetic variability of the Sardinian D. oxyrinchus populations could thus represent priority populations for conservation purposes, providing potential sources for recolonization in cases of local extinctions in other areas of the distribution range of the species.
6. When the sequences from Sardinia were compared with those available from other areas, the data seem to exclude the possibility that the Atlantic and Mediterranean host totally isolated populations or even different species, as recently suggested. However, additional markers and a larger sampling sites are needed to confirm these findings.

     

Spillover effects of a Mediterranean marine protected area on the European spiny lobster Palinurus elephas (Fabricius, 1787) resource

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Microsatellite variability in natural populations of the blackspot seabream Pagellus bogaraveo (Brünnick, 1768): a database to access parentage assignment in aquaculture

Eight microsatellite loci previously reported were assessed for their utility in parentage assignment in 96 individuals belonging to natural populations of the blackspot seabream Pagellus bogaraveo (Brünnick, 1768) from the Mediterranean and Northeast Atlantic regions. At the mtDNA level, no differentiation was found between these two regions but based on microsatellite data an overall discrete genetic differentiation is perceivable between the two regions separated by the Strait of Gibraltar. The number of alleles per locus ranged from 8 to 30. A database constructed with allele frequency data from six populations was used in a simulation parentage assignment test using the software cervus . The test showed that the number of markers used is enough to perform parentage assignments with real data. The polymorphic information content for each locus was very high (mean value of 0.849), with a total exclusionary power of 0.9995. In summary, seven of the eight microsatellites analysed proved to be sufficient and powerful tools for parentage assignment in hatcheries and the allele frequency data given here can be used to perform pedigree analysis against which real data may be tested.     

Implications of early selection for grain colour on iron and zinc content and productivity of common bean

Brazilian consumers prefer beans of the beige grain colour. Common beans help supply dietary requirements for iron and zinc. The objectives of this study were (i) to determine whether early selection for grain colour affects the iron and zinc content and the grain yield in common beans and (ii) if an association between these characteristics was found, to obtain lines of carioca‐type grains with a high iron and zinc content and good grain yield. We evaluated 96 F_3:4, F_3:5 and F_3:6 progeny at two locations for iron and zinc content and grain yield. We selected 48 progeny for grain colour and chose 48 others at random. We found that early selection for grain colour does not affect the zinc content and grain yield. A positive and high association was found between iron and zinc content, and both iron and zinc content were slightly negatively correlated with grain yield. It is possible to obtain common bean lines combining a high iron and zinc content with good grain yield as long as a selection index is used.     

Characterization of commercial kava-kava herbal drug and herbal drug preparations by means of nuclear magnetic resonance spectroscopy

The efficiency of one- and two-dimensional NMR experiments in characterizing the content of the constituents of both herbal drugs and herbal drug preparations is demonstrated for kava-kava. These experiments directly detect active constituents represented by kavalactones in both a finely powdered herbal drug and a commercial extract. In addition, NMR spectroscopy can detect all other compounds present in the extract. As previously evidenced, NMR experiments can represent a generally applicable technique for rapid screening and are a complement to the classical analytical techniques such as high-performance thin-layer chromatography, high-performance liquid chromatography, capillary gas chromatography, and electrophoresis. These experiments can be considered a very simple and fast analytical method to obtain a fingerprint of the herbal drugs and their preparations, and to quantify the content of the active principles of the extract.     

Sulfluramid volatility reduction by beta-cyclodextrin

Sulfluramid is an expensive active principle of insecticidal baits that is lost by volatilization during the pelletization of baits. To increase the thermal stability of sulfluramid, we tested its molecular encapsulation in beta-cyclodextrin (beta-CD), using molar ratios of 1:1 and 1:2 (sulfluramid:beta-CD), using the complex preparation techniques of coprecipitation and kneading. The physical mixture of sulfluramid and beta-CD was also tested for comparison. The products of complexation were characterized by differential scanning calorimetry, thermogravimetry, and derivative thermogravimetry, indicating the formation of a sulfluramid/beta-CD complex and showing that the release of the complexed sulfluramid occurs in the range of 270-300 degrees C, a temperature range that is well above the temperature at which sulfluramid sublimates (40 degrees C). This result warrants a reduced sulfluramid loss in the preparation of insecticidal baits. The preparation of the complex by kneading with molar ratio of 1:2 gave the highest yield of complex, about 64%, in relation to the theoretical maximum.     

Effects of elicitors on the production of resveratrol and viniferins in cell cultures of Vitis vinifera L. cv Italia

Methyl jasmonate, jasmonic acid and chitosan were tested as elicitors on cell suspension cultures obtained from Vitis vinifera cv Italia to investigate their effect on stilbene production. Stilbene accumulation in the callus, grown under nonelicited conditions, was also investigated. Calli and cell suspensions were obtained in a B5 culture medium supplemented with 0.2 mg L(-1) NAA and 1 mg L(-1) KIN. Stilbene determination was achieved by HPLC/DAD/MS. Whereas callus biosynthesized only piceid, cell suspensions elicited with jasmonates produced several stilbenes, mainly viniferins. In suspended cells, methyl jasmonate and jasmonic acid were the most effective in stimulating stilbene biosynthesis, whereas chitosan was less effective; in fact, the amount of stilbenes obtained with this elicitor was not significantly different from that obtained for the control cells. The maximum production of total stilbenes was at day 20 of culture with 0.970 and 1.023 mg g(-1) DW for MeJA and JA, respectively.