Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Evaluation of result variability with a commercial Johne's disease enzyme-linked immunosorbent assay kit and repeat testing of samples.

Two hundred one serum samples from individual dairy cows with a range of results on initial testing with a commercial Johne's disease enzyme-linked immunosorbent assay (ELISA) kit were repeat tested 5 times in each of 2 laboratories with kits produced by the same manufacturer. The results for the samples with all 10 replicates showed that the values for individual samples often had a coefficient of variation greater than 20%. As expected, the standard deviation for the results increased as the average value increased and the coefficient of variation was greater in samples with low mean values. The different lots of the commercial ELISA kit used in this study had a significant effect on both the optical density and the calculated sample to positive (S/P) ratio for test replicates. Based on the variability detected in S/P ratios of replicate samples, application of a single cutoff point to interpret individual test results as positive or negative for antibodies to Mycobacterium avium subspecies paratuberculosis could result in inconsistent classification of animals as positive or negative for Johne's disease. Such inconsistency in test interpretation leads to frustration in large animal veterinarians or producers trying to make management decisions based on individual test results. Instead of dichotomizing the test results as positive or negative based on a single cutoff value, reporting numerical values and supplying a classification scheme that includes a suspect category reflecting the uncertainty inherent in the test is recommended to provide more reliable result interpretation.     

Production of Haploid Sugar Beet (Beta vulgaris L.) by Culturing Unpollinated Ovules

The culture of unpollinated ovules is shown to be a suitable system for the production of haploid sugar beet (Beta vulgaris L.). The yield of haploids depended upon the genotype and varied between 0 and 13 % with a mean of 1.0 %. Haploid plants could be produced from approximately 50 % of all genotypes examined. The majority of the haploids isolated (about 90%) maintained the haploid genome level during the in vitro culture and propagation; 10% of the haploid clones showed a spontaneous doubling to the diploid genome level.     

Linkage and quantitative trait locus mapping of foliage late blight resistance in the wild species Solanum vernei

The global cultivation of potato (Solanum tuberosum) is threatened by epidemics caused by new variants of the late blight pathogen, Phytophthora infestans. New sources of durable late blight resistance are urgently needed and these may be found in wild Solanum species. The diploid wild species, S. vernei, has not previously been subjected to mapping of quantitative trait loci (QTLs) for late blight resistance. Two populations designated HGIHJS and HGG, originating from a cross between a clone of S. vernei and two different S. tuberosum clones were evaluated in field trials for late blight infestation. The relative area under the disease progress curve (RAUDPC) was estimated and used for QTL mapping. A linkage map of S. vernei, comprising 11 linkage groups, nine of which could be assigned to chromosomes, was constructed. Results indicated that the resistance in S. vernei was quantitatively inherited. Significant QTLs for late blight resistance were identified on chromosomes VIII (HGG), VI and IX (HGIHJS). In addition, potential QTLs were detected on chromosomes VII (HGIHJS) and IX (HGG). A putative and a significant QTL for tuber yield were found on chromosomes VI and VII in HGG, but no linkage between yield and resistance was indicated. The QTL for late blight resistance, which mapped to chromosome IX, could be useful for late blight resistance breeding as it was located close to the microsatellite marker STM1051 in both populations.     

Half-sib progeny evaluation and selection of potatoes resistant to the US8 genotype of Phytophthora infestans from crosses between resistant and susceptible parents

The objectives of this study were to evaluate the use of potato (Solanum tuberosum L.) late blight (Phytophthora infestans (Mont.) de Bary) resistant parents in cultivar development and identify superior clones possessing moderate to high late blight resistance combined with acceptable maturity and tuber quality. Ninety-five crosses were made between eight unadapted parents with reported late blight resistance (B0718-3, Bertita, Bzura, Greta, Libertas, Stobrawa, Tollocan and Zarevo) and susceptible parents (cultivars or advanced breeding clones) adapted to North American growing conditions. A total of 408 field selected clones were assessed for late blight resistance in the greenhouse and in the field using a mixture of US8 P. infestans isolates (A2 mating type, metalaxyl resistant) that overcame all known R-genes except R8 and R9. Clones with ≤ 10% infected foliar area in the greenhouse test or ≤ 0.30 RAUDPC (relative area under the disease progress curve) value in the field in 1998 were re-tested in 1999. A total of 118 (29% of 408) putative late blight resistant clones were selected. The eight late blight resistant parents differed in both the ability to transmit late blight resistance and in the level of resistance transmitted to the progeny. The Tollocan and B0718-3 families (half-sib progeny) had the greatest degree of resistance and frequency of resistant clones. Scott-Knott cluster analysis ranked 79 clones (67% of 118) in the high and moderate late blight resistant groups. Among these 79 clones, 19 clones had vine maturity equal to or earlier than mid-season combined with acceptable tuber quality. Further selection in 2000 resulted in eight advanced selected clones (six from Tollocan and two from B0718-3 families) with the same level of resistance as the parent combined with vine maturity and tuber quality equivalent to Atlantic, a standard cultivar for chip processing in North America. The results indicate that this breeding approach can be used to select parents for late blight resistance breeding and to identify superior clones with high levels of late blight resistance and marketable vine maturity and tuber quality. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of type of added salt and ionic strength on physicochemical and functional properties of casein isolates produced by electroacidification

A procedure developed for soybean protein precipitation which was based on electrodialysis was tested for the production of acid casein from reconstituted skim milk. In a previous paper, the performance of bipolar membrane electroacidification (BMEA) was evaluated under different conditions of ionic strength (micro(added) = 0, 0.25, 0.5, or 1.0 M) and added salt (CaCl(2), NaCl, or KCl) (1). The aim of this study, which is the complement of the work on evaluation of BMEA performance, was to evaluate the functionality of the protein isolates produced by BMEA and to compare the BMEA isolates to commercial isolates and an isolate produced by chemical acidification. It was not possible to show differences between the functional properties of isolates produced by BMEA, except at 1 M CaCl(2) micro(added), due to the variability of the isolates. However, the results showed that it is possible to obtain isolates similar to commercial isolates and that the addition of salt during the process does not induce variations in functional properties. From results on mineral concentrations, it appeared that the addition of monovalent cations did not influence the retention of monovalent or divalent cations in the BMEA isolates, while addition of divalent cations (CaCl(2)) influenced the retention of magnesium. According to previous results on evaluation of BMEA performances under different conditions of ionic strength and added salt, the difference observed for the BMEA isolate produced at 1.0 M CaCl(2) was confirmed.     

Patterns of amphibian diversity in Brazilian Amazonia: conservation implications

A literature survey was conducted to determine the amphibian diversity and distribution in Brazilian Amazonia. Patterns of endemisms and similarity of fauna between localities were also addressed. Twenty-eight inventories were found for the region, the majority localized in areas with easier access by road or river. A minimum of 163 amphibian species was recorded for Brazilian Amazonia. Although many species are endemic to the Amazonian lowlands as an entity, the patterns of species uniqueness among sites suggested low endemic distribution within the lowlands of the Amazon Basin. The mean similarity between localities varied from low to intermediate (mean=0.40), indicating that the Brazilian Amazonia is characterized by distinctive assemblages of amphibians throughout its extension. Localities further apart had lower similarity. No threatened species were recorded. These results contribute to determine priority areas for new inventories and establishment of conservation units. We suggest that areas next to the Amazon deforestation frontier should be prioritized for new studies due to the high rate of alteration and potential loss of species. Additionally, studies on amphibian population dynamics are few in Brazilian Amazonia and more of them should be emphasized to help to draw a better picture of the status of amphibians in this region.     

Aquatic Gastropods as Mercury Indicatorsin the Pantanal of Poconé Region (Mato Grosso, Brasil)

Gold mining is the principal polluting activity in themunicipality of Poconé at the northern limit ofthe Pantanal of Mato Grosso, Brasil and ischaracterized by the inadequate use of mercury in thegold extraction process. A way to contribute to theassessment of the environmental impact of mercuryrelease in the Pantanal is to make use of bioaccumulation. Thus, determinations of total mercuryin 188 Gastropods of three different species were madein seven different sampling sites during the dry andwet periods. Increase of mercury levels in the snailswas restricted to the gold mining area and adjacentsedimentation areas. The largest mercuryconcentrations were found at the Tanque dos Padressite, reaching 2.04 g g^-1 in individuals ofPomacea scalaris. In the other sampling sites,molluscs collected in the Araras/Piuval Bay complex,a mercury retention area, showed the largest mercuryconcentration, reaching up to 1.12 g g^-1.