Determination of F1 hybrid seed purity in pepper using PCR-based markers

We present the potential use of two kinds of PCR-based markers, RAPDs (Random Amplified Polymorphic DNA) and SPARs (Single Primer Amplification Reactions), as tools for hybrid seed purity determination. Five F₁ pepper hybrids (Capsicum annuum L.) and their parents were analyzed with 100 10-mer primers and 10 nucleotide repeat primers. We found at least one useful marker for testing the purity of all hybrids studied. Despite their dominant inheritance these markers could be an efficient implement in the process of quality testing of hybrid seeds. This revised version was published online in August 2006 with corrections to the Cover Date.     

Evidence of Migration and Endophytic Presence of Agrobacterium tumefaciens in Rose Plants

Agrobacterium tumefaciens was isolated from stem tumors of several rose cultivars showing that the bacterium is the causal agent of aerial galls in rose plants. No differences were observed in the characteristics of the Agrobacterium isolates from crown or aerial galls. Stem inoculation of ten rose cultivars showed that all of them were susceptible to A. tumefaciens but differences in the size of the resulting tumors were observed. The movement of A. tumefaciens in rose plants was demonstrated using two wild type strains and two antibiotic resistant mutants. Three months after inoculation, the inoculated strains were recovered in the roots, crown and below and above the inoculation site but low numbers of pathogenic Agrobacterium cells were isolated. New tumors appeared in 5% of the noninoculated wounds. A. tumefaciens was isolated from the stem at different distances from the tumor in naturally infected plants. In symptomless commercial plants, the isolation from the roots, crown and at different stem levels demonstrated the existence of systemic and latent infections in rose. Direct isolation using a nonselective and selective media with or without a previous enrichment step were efficient methods for isolating tumorigenic Agrobacterium from the different parts of rose plants.     

Inter‐annual variability in amphibian assemblages: implications for diversity assessment and conservation

• 1. Diversity assessments and conservation management should take into account the dynamic nature of populations and communities, particularly when they are subject to highly variable and unpredictable environmental conditions.
• 2. This study evaluates the inter‐annual variability in the assemblage composition (temporal turnover) of an amphibian community breeding in a highly dynamic habitat, a Mediterranean temporary pond system, during a 4‐year period.
• 3. A comprehensive framework is provided to evaluate temporal turnover from data of a differing nature (species richness, presence/absence and relative abundance) and, especially, to discern variation in richness (species loss) from changes in the identity or abundance of species (species replacement).
• 4. Results show that the pond amphibian assemblages in Doñana National Park exhibited high inter‐annual variability during the study period, both in the number of species, species identity and their relative abundance. This result provides evidence for the inadequacy of surveys conducted only in one breeding season to characterize the species assemblage associated with a given pond. Besides, it suggests that a given pond offers different breeding opportunities over time, being suitable for different species depending on the year. This alternation will contribute to the medium‐term preservation of all species in the assemblage.
• 5. It is highly relevant to preserve the natural dynamism and spatial variability of temporary pond systems, which will favour the conservation of populations through their intrinsic variability. Copyright © 2010 John Wiley & Sons, Ltd.

     

Marine and freshwater crab meals in diets for red porgy (Pagrus pagrus): effect on growth,fish composition and skin colour

River crab (RC) meal (Procambarus clarkii) and marine crab (MC) meal (Chaceon affinis) were tested as a partial replacement for fish meal in diets for red porgy (Pagrus pagrus), and their effects on growth performance, fish proximate composition and skin colouration were evaluated. Red porgy were fed during 165 days with five diets. High‐quality fish meal diet was used as a control diet (CD). Protein of fish meal in the control was replaced by increasing the dietary levels of protein derived from RC and MC by up to 10% and 20% of each of them (RC10, RC20, MC10 and MC20). Fish fed on MC20 showed the highest values in feed intake, weight gain and growth (%). No differences were found in FCR and protein efficiency ratio among the treatments. Inclusion of both crab meals in diets significantly decreased the lipid content in whole fish compared with the control animals. On the other hand, no differences in muscle composition were found between the diets. Feeding both crab meals resulted in colour improvement compared with that of the control fish, with better hue values for the RC meal group than those for the MC meal group. The crab meals tested in the present study are suitable as a partial replacement for fish meal in diets for the red porgy, with the MC meal improving growth and both crabs meals improving skin colour, with further improvements in skin colour produced in fish‐fed diets containing the RC meal.     

Evaluation of Agave lechuguilla by‐product crude extract as a feed additive for juvenile shrimp Litopenaeus vannamei

Agave lechuguilla is a succulent plant species, mainly distributed in the northeast of Mexico and south of the United States of America. The main use for this plant is the fibre´s extraction (known as Tampico fibres), resulting in 15% of fibres and 85% of a by‐product waste named guishe. The lechuguilla collectors, normally incinerate the guishe, thus causing environmental contamination. Interestingly, recent studies showed that guishe contains molecules with nutritional properties, such as saponins, flavonoids and sugars. Therefore, in this work, we evaluated the effect of the crude extract of guishe as a feed additive in whiteleg shrimp diets. According to that, MS‐HPLC analysis of the extract showed the presence of saponins such as diosgenin, smilagenin, hecogenin, manogenin, tigogenin hexose, yucagenin, chlorogenin, diosgenin diglucoside and the flavonol, quercetin. After chemical analysis, the crude extract was included into an experimental diet in four levels; 0% (L0%), 0.1% (L0.1%), 0.3% (L0.3%) and 0.6% (L0.6%). Dietary incorporation of the extract was evaluated by zootechnical performance, haemolymph biochemistry, histomorphology and digestive enzyme activity of shrimps. After 5‐week feeding, the L0.3% diet showed significantly higher growth and better feed utilization among treatments. A significant increase in tubule epithelium height and tubule coverage area from hepatopancreas in shrimp under L0.3% diet compared with the control diet suggest an improvement of the health and nutritional status of the shrimp. Inclusion of L0.3% and L0.6% of the crude extract resulted in a reduction in amylase activity, without effect in glucose levels in the haemolymph. Thus, we suggest that lechuguilla guishe crude extract contains nutritional molecules that may be used as a feed additive to promote shrimp productivity.     

Local and remote physical processes driving variability of the planktonic system in the Juan Fernández Archipelago: A multidisciplinary framework providing conservation insights

1. The Juan Fernandez Ridge, a vulnerable marine ecosystem located far off the coast of central Chile and formed by several seamounts, guyots and three islands (Robinson Crusoe, Santa Clara and Alejandro Selkirk), has recently been declared a Coastal Marine Protected Area of Multiple Uses with several National Parks embedded in it.
2. Recent studies have highlighted the influence of remote and local oceanographic structures on the hydrographic dynamics of this ridge. However, there is still a gap in understanding how they affect the structure and dynamics of the surrounding insular planktonic communities.
3. A hydroacoustic and oceanographic survey was conducted during the austral spring (October 2016), including hydrographic and zooplankton sampling around Robinson Crusoe Island. Oceanographic features were identified and tracked using satellite data (chlorophyll-a (Chl-a) and sea surface temperature) and modelling results.
4. Two events of Chl-a increase relative to a threshold (>0.45 mg m⁻³) were forced by different physical processes, both affecting the western side of Robinson Crusoe Island. In event A during the cruise period, Chl-a subsurface maxima were associated with the arrival of a coastal meander originating on the continental shelf off Chile (remote process); the zooplankton was dominated by copepods and salps, with an evident coastal–oceanic gradient. In event B, Chl-a maxima were linked to a local upwelling forced by the intensification of a localized SSW wind. No influence of remote eddies or local Von Kármán vortices on Chl-a distribution was observed.
5. These findings highlight the influence of remote and local physical processes on the structure of planktonic communities around Robinson Crusoe Island. Understanding the variability of these mechanisms and their effects at the base of the pelagic food web is critical in adopting an ecosystem-level approach.

     

Diterpenes from Salvia broussonetii transformed roots and their insecticidal activity

The new diterpenes brussonol (1) and iguestol (6alpha,11-dihydroxy-12-methoxy-abieta-8,11,13-triene) (2) with an icetexane and a dehydroabietane skeleton, respectively, have been isolated from hairy root cultures of Salvia broussonetii. Other previously known diterpenes, 7-oxodehydroabietane, 11-hydroxy-12-methoxyabietatriene, taxodione, inuroyleanol, ferruginol, deoxocarnosol 12-methyl ether, cryptojaponol, pisiferal, sugiol, isomanool, 14-deoxycoleon U, 6alpha-hydroxydemethylcryptojaponol, demethylsalvicanol, and demethylcryptojaponol, were also obtained from these roots. The insect antifeedant and toxic effects of several of these compounds were investigated against the insect pests Spodoptera littoralis and Leptinotarsa decemlineata. Additionally, their comparative cytotoxic effects were tested on insect Sf9 and mammalian CHO cells. Demethylsalvicanol (4) was a moderate antifeedant to L. decemlineata, whereas brussonol (1) was inactive. 14-Deoxycoleon U (15) was the strongest antifeedant, whereas demethylcryptojaponol (11) was toxic to this insect. None of these compounds had antifeedant or negative effects on S. littoralis ingestion or weight gains after oral administration. Demethylcryptojaponol (11) was cytotoxic to mammalian CHO and insect Sf9 cell lines, followed by the icetexane derivative brussonol (1), with moderate cytotoxicity in both cases. The remainder of the test compounds showed a strong selective cytotoxicty to insect Sf9 cells, with demethylsalvicanol (4) being the most active.     

Characterization and growth conditions of Vibrio parahaemolyticus strains with different virulence degrees that cause acute hepatopancreatic necrosis disease in Litopenaeus vannamei

The phenotypic characteristics and growth kinetics at several temperatures, salinities, and pH values of three Vibrio parahaemolyticus (Vp) strains with different virulence and one nonpathogenic strain were evaluated. Independent of the virulence of the strain, a high metabolic diversity was found, which yielded different colored phenotypes on the CHROMagar? Vibrio. All strains were resistant to ampicillin and carbenicillin, and Vp AHPND+ organisms were the most sensitive to enrofloxacin. The exponential growth of Vp strains started at 1–2 hr of incubation, although no relationship was observed between the bacterial density and degree of virulence. Moreover, the growth of the most virulent strain was independent of the nutrients in the incubation media during the initial hour postinoculation. No strain grew at 4°C in 0% NaCl and pH 4, but only Vp AHPND+ grew at 44°C. For all strains, the lag phase was proportional to the NaCl concentration, and the growth was better at pH 8–9. However, the Vp AHPND? strain displayed a greater variability, was more sensitive to extreme conditions, and showed a lag phase of 9 hr independent of the pH.     

Evaluation of single and comparative intradermal tuberculin tests for tuberculosis eradication in caprine flocks in Castilla y León (Spain)

Goats can act as reservoirs for tuberculosis (TB) infection. The main etiological agents of TB in goats are Mycobacterium caprae and Mycobacterium bovis and they infect also a wide range of domestic and wild animals and humans. Control programmes based mainly on the application of single and comparative intradermal tuberculin (SIT and SCIT respectively) tests are being implemented in certain regions of Spain with a high density of caprine flocks as Castilla y León, including goats with epidemiological relationship with cattle. The aim of this study was to evaluate the performance of the intradermal tests in naturally TB-infected caprine flocks from this region. The study was performed using data from 17,450 goats in 54 different flocks that were classified as TB-infected in the control programmes executed in 2010 and 2011. Data from 1237 goats from 7 dairy flocks depopulated after the first intradermal testing were used to estimate the sensitivity (Se) using bacteriology as the gold-standard. Overall Se of the SIT test using the severe interpretation was 43.9% (CI 95%, 40.4–47.4) and decreased to 38.8% (CI 95%, 35.5–42.3) using the standard interpretation. Overall Se of the SCIT test ranged between 21.3% (CI 95%, 17.6–25.4) and 7% (CI 95%, 4.9–9.8) depending of the interpretation criteria. A significant weak positive correlation was found between age and skin fold thickness (Spearman’s test p < 0.05). Results from this study yielded, in general, low Se values probably due the systematic detection and slaughter of reactors as a consequence of the eradication programme in previous years or the presence of factors that may interfere in the diagnosis. Therefore, these results suggest the necessity of including ancillary diagnostic tools and/or strict interpretation criteria to maximize detection of positive animals in infected settings.     

Utilization of enzyme mixtures to retard bread crumb firming

The influence of enzyme mixtures containing amylase and lipase activities on straight dough bread staling was studied. Amylopectin retrogradation, crumb firming, amylose-lipid complexes, and dextrin production were analyzed in bread samples supplemented with two enzyme mixtures. The addition of enzyme mixtures to bread formula causes a beneficial effect on bread keeping properties and the formation of a more thermostable amylose-lipid complex than the one found in control bread. Amylopectin retrogradation was inhibited by the use of the enzyme; the effect was accompanied by reduced crumb-firming rates. The enzymatically generated water-soluble dextrins (maltose and DP3, DP4, DP5, and DP6 dextrins) are the most effective in preserving crumb softness during bread storage.