Effects of exogenous amylase on the in vitro digestion kinetics of whole-crop maize silages made from flint or dent grain type at different phenological stages grown in tropical condition

The effect of exogenous amylase on the in vitro rumen digestion kinetics of whole-crop maize silage made from dent (RB9004) or flint grain type (RB9308) was evaluated at different phenological stages: soft dough (SOD), early dent (EAD), ½ milkline (½M) and ¾ milkline (¾M). Forage was harvested from 70 to 110 days after sowing. Two rumen-cannulated cows receiving or not exogenous amylase (0.7 g/kg dry matter—DM, provided to achieve 396 kilo Novo units of amylase activity/kg of TMR DM) were used as donor of ruminal fluid. The in vitro gas production kinetics was evaluated according to a dual-pool logistic model. The chemical composition and gas production kinetics were affected by the hybrid and phenological stages. The flint hybrid had lower range for chemical analysis among physiological stages. Harvesting at ½M and ¾M improved DM content, bromatological composition and silage quality parameters compared to dent or flint types. Amylase (i) increased methane (CH₄) production and in vitro dry matter digestibility (IVDMD) in ½M stage, (ii) improved digestion kinetics by reducing lag time and increasing total gas production and fermentation rates of non-fibrous carbohydrates (NFC) and fibrous carbohydrates (FC), and (iii) increased extent and fermentation rate of NFC and increased fermentation rate of FC fraction in whole-crop maize silages produced from dent or flint types in all phenological stages. Harvesting between ½M and ¾M is the best phenological stage to improve chemical composition and silage quality parameters. Exogenous amylase showed improvements on fibre digestion of silages at ½M and ¾M phenological stages in both grain types of corn.     

An HPLC investigation of flower colour and breeding of anthocyanins in species and hybrids of Alstroemeria

The tepals of 28 Chilean species of Alstroemeria and 183 interspecific hybrids were analysed for anthocyanin content by high-performance liquid chromatography (HPLC). The anthocyanins were identified as 3-rutinosides of 6-hydroxydelphinidin, 6-hydroxyeyanidin, cyanidin, and delphinidin and 3-glyeosides of cyanidin and delphinidin, some of which were acylated with malonic acid. Comparisons of the anthocyanin contents in parents and offspring showed that no anthocyanidin or acylation pattern was dominant, and that offspring values were close to mid-parent values for the percentage of malonated anthocyanins, whereas the inheritance of cyanidin, 6-hydroxycyanidin, and delphinidin seems more complicated. Flower colour, hue, and intensity were measured by CIELab in fresh tepals and compared with their anthocyanin content and the estimated flavonoid concentrations. Colour intensity was positively correlated with anthocyanin concentration. Compared with flowers containing exclusively cyanidin 3-glycosides, the hues of flowers with delphinidin 3-glycosides were bluer and with 6-hydroxycyanidin 3-glycosides redder, respectively. Both malonation of anthocyanin and co-pigmentation with flavonoids caused a shift to bluish hues, irrespective of the anthocyanidins. By quantifying both chemical and colorimetric characteristics a model for the effect of anthocyanin on Alstroemeria flower colour was established. Breeding of new cultivars of Alstroemeria is discussed.     

Alterations in the brain monoaminergic neurotransmitters of rainbow trout related to naphthalene exposure at the beginning of vitellogenesis

The contents of dopamine (DA), noradrenaline (NA), serotonin (5HT), and some related metabolites were studied in different brain regions of rainbow trout at two different stages of sexual maturation (at the beginning of vitellogenesis), after naphthalene (NAP) administration. The effects of NAP varied according to duration of exposure, brain region and vitellogenesis stage of the trout, and were more significant during previtellogenesis. The changes observed in DA metabolism were generally stimulatory after exposure for 3 h, and either stimulatory or inhibitory (depending on the brain regions) after exposure for 3 days to NAP. NA levels were altered by NAP in various brain regions, but only during previtellogenesis. With respect to 5HT, treatment with NAP reduced levels of the amine and/or its main metabolite in most of the brain regions studied, particularly 3 h after treatment. The results suggest that NAP might interfere with the processes regulating brain monoamine metabolism, either locally or indirectly by altering steroid feedback to brain centres, and thus disrupt endocrine control of reproductive development through the brain–pituitary axis.     

Spasmolytic Effect of Caulerpine Involves Blockade of Ca2+ Influx on Guinea Pig Ileum

In this work, we investigated the spasmolytic effect of caulerpine, a bisindole alkaloid isolated from marine algae of the Caulerpa genus, on guinea pig ileum. Our findings indicated that caulerpine inhibited phasic contractions induced by carbachol (IC₅₀ = 7.0 ± 1.9 × 10⁻⁵ M), histamine (IC₅₀ = 1.3 ± 0.3 × 10⁻⁴ M) and serotonin (IC₅₀ = 8.0 ± 1.4 × 10⁻⁵ M) in a non-selective manner. Furthermore, caulerpine concentration-dependently inhibited serotonin-induced cumulative contractions (pD′₂ = 4.48 ± 0.08), shifting the curves to the right with E_max reduction and slope of 2.44 ± 0.21, suggesting a noncompetitive antagonism pseudo-irreversible. The alkaloid also relaxed the ileum pre-contracted by KCl (EC₅₀ = 9.0 ± 0.9 × 10⁻⁵ M) and carbachol (EC₅₀ = 4.6 ± 0.7 × 10⁻⁵ M) in a concentration-dependent manner. This effect was probably due to inhibition of Ca²⁺ influx through voltage-gated calcium channels (Ca_V), since caulerpine slightly inhibited the CaCl₂-induced contractions in depolarizing medium without Ca²⁺, shifting the curves to the right and with E_max reduction. According to these results, the spasmolytic effect of caulerpine on guinea pig ileum seems to involve inhibition of Ca²⁺ influx through Ca_V. However, other mechanisms are not discarded.     

True hermaphroditism in a horned goat with 60XX/60XY chimerism

Anatomical, behavioral, histological, endocrinological, and cytogenetic characteristics were determined in a horned intersex goat. Histology of the gonads confirmed that the goat was a true hermaphrodite. Cytogenetic analysis showed that it was a chimera (60XX/60XY). Use of laparoscopy allowed determination of characteristics of the internal gonads.     

Genetic typing of classical swine fever virus

Three regions of the classical swine fever virus (CSFV) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. Sequence data-sets were assembled for 55 CSFVs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the E2 envelope glycoprotein gene and 409 nucleotides of the NS5B polymerase gene. Phylogenetic analysis of each data-set revealed similar groups and subgroups. For closely related viruses, the more variable or larger data-sets gave better discrimination, and the most reliable classification was obtained with sequence data from the NS5B region. No evidence was found for intertypic recombination between CSFVs. A larger data-set was also analysed comprising 190 nucleotides of E2 sequence from 100 CSFVs from different parts of the world, in order to assess the extent and global distribution of CSFV diversity. Additional groups of CSFV are evident from Asia and the nomenclature of Lowings et al. (1996) [Lowings, P., Ibata, G., Needham, J., Paton, D., 1996. J. Gen. Virol. 77, 1311-1321] needs to be updated to accommodate these. A tentative assignment, adapting rather than overturning the previous nomenclature divides CSF viruses into three groups with three or four subgroups: 1.1, 1.2, 1.3; 2.1, 2.2, 2.3; 3.1, 3.2, 3.3, 3.4. The expanding data-base of CSFV sequences should improve the prospects of disease tracing in the future, and provide a basis for a standardised approach to ensure that results from different laboratories are comparable.     

Recruitment of catarina scallop (Argopecten ventricosus) larvae on artificial collectors off the NE coast of the Gulf of California

We evaluated recruitment of larvae of catarina scallop, Argopecten ventricosus, in the area of Puerto Peñasco, NE Gulf of California. We moored artificial collectors in six sites from June 2007 to August 2008 and replaced them every 2 months. We used monthly (July 2002–September 2011) sea surface temperature (SST, °C) and surface chlorophyll‐a concentration (SSChl, mg m^?3) Aqua/MODIS satellite data to describe seasonal environmental behaviour study area. Also, we recorded bottom temperature at each site every 4 h, and every 2 months measured sea surface salinity, temperature and dissolved oxygen. We used a repeated measures anova to evaluate differences in the number of recruited spat between main factors, and analysed the presence of multimodal spat shell size frequency distributions. Overall, spat recruitment was negatively correlated with seawater temperature and showed higher spat recruitment abundances throughout winter, which is the season with the highest surface chlorophyll a concentration. We estimated multimodal shell size frequency distributions characterized by more than one modal size. The natural collection of A. ventricosus spat on artificial collectors in the area can be successfully performed over a protracted period (November–December to May–June). Our results extend the area where collection of A. ventricosus spat can be successful.     

Beta-carotene isomer composition of sub- and supercritical carbon dioxide extracts. Antioxidant activity measurement

In the present work sub- and supercritical extraction conditions using carbon dioxide were studied in order to obtain extracts with different compositions from the green microalgae Dunaliella salina. Different compositions of beta-carotene isomers were identified in the extracts by using HPLC-DAD. Also, antioxidant activity of the extracts was measured using a TEAC assay. An experimental design was applied considering two factors, extraction pressure and temperature, in a wide range of values, trying to maximize the extraction yield. Higher yields were obtained at high pressures and low temperatures, that is, at higher CO2 densities. Attempts were made to correlate the antioxidant activity of the extracts with their chemical composition by means of principal component analysis. A certain relationship was found between their antioxidant activity and the isomeric composition of beta-carotenes. As a result, an original equation is proposed to predict the antioxidant activity of extracts from D. salina in terms of the ratio 9-cis-beta-carotene/all-trans-beta-carotene, the concentration of alpha-carotene, and, especially, the concentration of 9-cis-beta-carotene.     

Tagging and mapping a second resistance gene for <Emphasis Type="Italic">Fusarium</Emphasis> wilt race 0 in chickpea

A second gene conferring resistance to the chickpea wilt pathogen, Fusarium oxysporum f. sp ciceris race 0, has been mapped to linkage group 2 (LG2) of the chickpea genetic map. Resistance to race 0 is controlled by two genes which segregate independently; one present in accession JG62 (Foc0 ₁ /foc0 ₁ ) and mapping to LG5 and the second present in accession CA2139 (Foc0 ₂ /foc0 ₂ ) but remaining unmapped. Both genes separately confer complete resistance to race 0 of the wilt pathogen. Using a Recombinant Inbred Line (RIL) population that segregated for both genes (CA2139 × JG62) and the genotypic information provided by two markers flanking Foc0 ₁ /foc0 ₁ ten resistant lines containing the resistant allele Foc0 ₂ /foc0 ₂ were selected. Genotypic analysis using these ten resistant lines paired with ten susceptible RILs, selected in the same population, revealed that sequence tagged microsatellite sites (STMS) markers sited on LG2 were strongly associated with Foc0 ₂ /foc0 ₂ . Linkage analysis, using data from two mapping populations (CA2139/JG62 and CA2156/JG62), located Foc0 ₂ /foc0 ₂ in a region where genes for resistance to wilt races 1, 2, 3, 4 and 5 have previously been reported and which is highly saturated with tightly-linked STMS markers that could be used in marker-assisted selection (MAS).     

Evidence for a relationship between bovine erythrocyte lipid membrane peculiarities and immune pressure from ruminal ciliates

Erythrocytes of bovines and other ruminants have a strikingly anomalous phospholipid composition, with low or absent phosphatidylcholine (PC) together with high sphingomyelin (SM) content. Here, we report the presence in normal bovine serum of high levels of anti-phospholipid antibodies of IgM isotype against, PC and the phosphono analogue of phosphatidylethanolamine, aminoethylphosphonolipid (AEPL), normally produced by rumen ciliates. In contrast, no antibodies were detected against SM or N-acyl-phosphatidylethanolamine (NAPE), the major components of bovine erythrocytes. In addition, we found that exposure of the ciliate Tetrahymena thermophila to bovine serum results in rapid lysis, an effect that was inhibited by adsorption of the serum with SM/AEPL liposomes. Furthermore, incubation with bovine serum had a similar effect on freshly obtained ruminal ciliates, and the lytic activity was eliminated by pre-adsorption of the serum with SM/PE liposomes. The ruminant mode of life with its concomitant ciliate fauna is hereby linked to the peculiar conformation of bovine erythrocyte membranes. We propose that the unique phospholipid composition of bovine erythrocytes appears as an evolutionary adaptation to tolerate the lytic effects of anti-phospholipid antibodies generated against AEPL, a membrane component of the huge mass of ruminal ciliates, necessary commensals of this group of mammals.