Assay performance during validation of freezing channel catfish Ictalurus punctatus (Rafinesque) infected with a Gram‐negative bacterium

Recovery of bacteria from infected fish during population sampling can be affected by factors including the type of assay, method of specimen preservation and concentration of bacteria present. Consequently, before use in field sampling, methods should be validated. The three objectives of this study were, first, to determine whether a channel catfish Ictalurus punctatus (Rafinesque) fingerling classified as positive for Gram‐negative Edwardsiella ictaluri infection according to bacterial culture before freezing was also classified as positive after freezing, second, to determine how direct culture from the kidney (DIRECT), culture of homogenate (HOMOG) and standard PCR (PCR) agree with bacterial culture in terms of classifying fish as positive or negative and third, to estimate diagnostic sensitivity (dSe) and diagnostic specificity (dSp) for DIRECT, HOMOG and PCR. In fresh and frozen fish, as bacterial concentration decreased, the ability of each assay to detect positive fish also decreased, especially when there were <10⁴ colony‐forming units per gram (CFU g^?1) tissue. HOMOG proved to be the most reliable at correctly classifying catfish, whether they were subclinically or clinically infected. PCR assay was the least reliable. Overall, values for this study population for dSe were 0.66, 0.92 and 0.43, and for dSp were 0.86, 0.91 and 0.95, for DIRECT, HOMOG and PCR respectively.     

Organization and sequence of four flagellin-encoding genes of Edwardsiella ictaluri

Edwardsiella ictaluri , the cause of enteric septicaemia in channel catfish ( Ictalurus punctatus ), is motile by means of peritrichous flagella. We determined the complete flagellin gene sequences and their organization in E. ictaluri by sequencing genomic segments from a λ-ZAP phage genomic library of E. ictaluri . Four flagellin genes ( fliC1, fliC2, fliC3 and fliC4 ) are arranged in tandem within 6 kb in the E. ictaluri genome. Each flagellin-coding sequence is preceded by a σ²⁸ recognition site consensus sequence. The predicted amino acid sequences of all four flagellin proteins (between 36 and 37.5 kDa) are similar in the N-terminal (1–160 aa) and C-terminal (last 74 aa) portions and are divergent in the central portion of the proteins. Proteins encoded by flC1, fliC2 and fliC3 are more similar to each other (88–90% aa identity) than to the protein encoded by fliC4 (76–78% aa identity). basic local alignment search tool analysis of GenBank sequences showed that all flagellin aa sequences are more similar to those of Serratia marcescens (72–74% identity) than to those of Edwardsiella tarda (≤64% identity). Primary determination of E. ictaluri flagellin gene sequences facilitate advanced studies on the role of flagella in host–pathogen interaction.     

Growth Response and Resistance to Streptococcus iniae of Nile Tilapia, Oreochromis niloticus, Fed Diets Containing Distiller's Dried Grains with Solubles

This study was conducted to evaluate the effect of dietary levels of distiller’s dried grains with solubles (DDGS) on growth, body composition, hematology, and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus iniae challenge. Five isocaloric diets containing DDGS at levels of 0, 10, 20, and 40%, and 40% DDGS + lysine (Diets 1–5) as partial replacements of a combination of soybean meal (SBM) and corn meal (CM) on an equal protein basis were fed to juvenile Nile tilapia (9.41 ± 0.14 g) for 10 wk. Fish fed Diet 4 had the lowest weight gain (WG), feed efficiency ratio, protein efficiency ratio (PER), and whole‐body protein. Supplementation of lysine to the 40% DDGS diet (Diet 5) improved WG and PER. Hematological and immunological parameters were not affected by dietary treatment. There were no significant differences among the average number of days to first mortality after S. iniae challenge and cumulative mortality 14 d postchallenge among fish in various treatments. DDGS can be incorporated in tilapia diet at a level of 20% as a substitute for a combination of SBM and CM without affecting their growth performance, body composition, hematological parameters, immune response, and resistance to S. iniae infection.     

Quantifying contributions to tournament catches among resident,stocked and hybrid black basses (Micropterus spp.)

Millions of Florida bass, Micropterus floridanus Lesueur, are stocked annually into populations of largemouth bass, Micropterus salmoides Lacepède, to increase trophy fish abundance. However, little effort has related the role that resultant hybrids make to angler catches. Largemouth bass were sampled from an important recreational fishery subject to extensive Florida bass stocking to address the hypothesis that anglers capture Florida bass, largemouth bass and hybrids at rates equivalent to their overall abundance in the population. Fin clips obtained from tournament angling events (n = 348) and boat‐mounted electrofishing sampling (n = 219) were screened at 38 species‐diagnostic markers and individuals were assigned to genealogical classes using Bayesian clustering algorithms. No significant differences were identified between angler and electrofishing catches providing evidence that hybridised individuals stemming from a long‐term stocking programme may constitute an important contribution to tournament angling catch.     

The 15N-isotope dilution technique applied to the estimation of biological nitrogen fixation associated with Paspalum notatum cv. batatais in the field

The contribution of associated biological nitrogen fixation to the nitrogen nutrition of Paspaulum notatum cv. batatais was estimated using the ¹⁵N-isotope dilution technique. The plants were grown in the field in concrete cylinders (60 cm dia) filled with soil, to which were added small quantities of ¹⁵N-labelled fertilizer at frequent intervals over 12 months. The pensacola cultivar of P. notatum was used as a non-N₂-fixing control plant. This was justified by the observation that nitrogenase (intact core C₂H₂ reduction) activity associated with the pensacola cultivar was consistently much lower than that of the batatais cultivar.At the first harvest, no evidence for N₂ fixation associated with the batatais cultivar was obtained, probably because of slow establishment of the N₂-fixing association. However, at the subsequent three harvests the batatais cultivar exhibited a lower ¹⁵N-enrichment and yielded more N than the pensacola cultivar. These data together suggested that 8–25% of the N in the batatais cultivar originated from N₂ fixation.The grass Paspalum maritimum was also included in the experiment and exhibited low nitrogenase activity similar to that of the pensacola cultivar of P. notatum. However, the total N and ¹⁵N data of these two grasses were not in good agreement indicating that it is important for the use of the isotope dilution technique that control plants are of very similar physiology and growth habit.     

Population management: potential impacts of advances in genomics

Even with advanced gene technologies continued population improvement remains a key foundation for future genetic gain in forest trees. Currently, this is served by maintaining genetic diversity while capturing genetic improvement, often through structuring populations into a genetic hierarchy, setting population sizes, and managing pedigrees and inbreeding. In the future, information from genomic technologies will complement classical approaches, such as common-garden field experiments, for characterizing the genetic base and measuring and monitoring genetic diversity. This will entail directly measuring DNA sequence diversity of both selectively neutral markers and functional genes. Calibrating neutral marker diversity against functional diversity will become easier with functional genomics. For maintaining pedigree and managing inbreeding, genomic information can be used to relax some traditional tenets of population management. With future knowledge of functional polymorphisms, the better understanding of the nature and origins of genetic variation should enhance management of populations to both conserve genetic diversity and exploit it by more efficient selection. Where fungal diseases threaten biotic crises, very large populations may be needed, the requisite sizes often being very uncertain. Gene discovery holds enormous promise, but depends heavily on comparative genomics, capitalizing on genomic information from Arabidopsis, Populus and Eucalyptus, and the increasing numbers of accessible ESTs. Much greater insights into non-codon mutational processes and rates should also guide population management. A key challenge, however, will be to utilize information and apply tools cost-effectively. Also, very detailed genomic information, exemplified by the poplar-genome sequencing, may allow earlier adaptation of technology and development of new information in angiosperms than in gymnosperms.     

Release of acetaldehyde from beta-cyclodextrins inhibits postharvest decay fungi in vitro

Many naturally occurring plant volatiles are known to have antifungal properties. However, they have limited use because they diffuse rapidly in air. In this in vitro study, acetaldehyde was chosen as a prototype volatile in order to study the controlled release of antifungal volatiles from cyclodextrins (CD). The major postharvest pathogens Alternaria alternata, Botrytis cinerea, and Colletotrichum acutatum were exposed to the pure volatile for 7 days at 23 degrees C. Acetaldehyde was most effective against A. alternata, followed by C. acutatum, and B. cinerea, with 0.12, 0.56, and 1.72 microL/L in air being required to inhibit fungal growth, respectively, according to the bioassay developed. Second, the effectiveness of the new beta-CD-acetaldehyde release system was evaluated against A. alternata for 7 days at 23 degrees C. Sufficient volatile was released from 0.7 g of beta-CD-acetaldehyde to prevent fungal growth in vitro.