Triploid grass carp susceptibility and potential for disease transfer when used to control aquatic vegetation in reservoirs with avian vacuolar myelinopathy.

Avian vacuolar myelinopathy (AVM) is an often-lethal neurologic disease that affects waterbirds and their avian predators (i.e., bald eagles Haliaeetus leucocephalus) in the southern United States. Feeding trials and field surveys provided evidence that AVM is caused by a toxin-producing, undescribed cyanobacterium (UCB), which grows as an epiphyte on the leaves of submerged aquatic vegetation (SAV). Reservoirs with documented AVM epornitics support dense growth of nonnative SAV. Waterbirds ingest the toxin when feeding on aquatic plants with the epiphytic UCB, and secondary intoxication occurs when raptors consume these birds. Vegetation management has been proposed as a means to reduce waterbird exposure to the putative toxin. We fed aquatic vegetation with and without the UCB to triploid Grass Carp Ctenopharyngodon idella in laboratory and field trials. Only Grass Carp that ingested aquatic vegetation with the UCB developed lesions in the central nervous system. The lesions (viewed using light microscopy) appeared similar to those in birds diagnosed with AVM. Grass Carp that received aquatic vegetation without the UCB were unaffected. Grass Carp tissues from each treatment were fed to domestic chickens Gallus domesticus (an appropriate laboratory model for AVM) in a laboratory trial; the chickens displayed no neurologic signs, and histology revealed a lack of the diagnostic lesions in brain tissues. Results from our trials suggest that (1) triploid Grass Carp are susceptible to the AVM toxin, although no fish mortalities were documented; and (2) the toxin was not accumulated in Grass Carp tissues, and the risk to piscivorous avifauna is likely low. However, a longer exposure time and analysis of sublethal effects may be prudent to further evaluate the efficacy and risk of using triploid Grass Carp to manage aquatic vegetation in a system with frequent AVM outbreaks.

Towards the ictalurid catfish transcriptome: generation and analysis of 31,215 catfish ESTs.

BACKGROUND: EST sequencing is one of the most efficient means for gene discovery and molecular marker development, and can be additionally utilized in both comparative genome analysis and evaluation of gene duplications. While much progress has been made in catfish genomics, large-scale EST resources have been lacking. The objectives of this project were to construct primary cDNA libraries, to conduct initial EST sequencing to generate catfish EST resources, and to obtain baseline information about highly expressed genes in various catfish organs to provide a guide for the production of normalized and subtracted cDNA libraries for large-scale transcriptome analysis in catfish. RESULTS: A total of 17 cDNA libraries were constructed including 12 from channel catfish (Ictalurus punctatus) and 5 from blue catfish (I. furcatus). A total of 31,215 ESTs, with average length of 778 bp, were generated including 20,451 from the channel catfish and 10,764 from blue catfish. Cluster analysis indicated that 73% of channel catfish and 67% of blue catfish ESTs were unique within the project. Over 53% and 50% of the channel catfish and blue catfish ESTs, respectively, had significant similarities to known genes. All ESTs have been deposited in GenBank. Evaluation of the catfish EST resources demonstrated their potential for molecular marker development, comparative genome analysis, and evaluation of ancient and recent gene duplications. Subtraction of abundantly expressed genes in a variety of catfish tissues, identified here, will allow the production of low-redundancy libraries for in-depth sequencing. CONCLUSION: The sequencing of 31,215 ESTs from channel catfish and blue catfish has significantly increased the EST resources in catfish. The EST resources should provide the potential for microarray development, polymorphic marker identification, mapping, and comparative genome analysis.

First report of Yersinia ruckeri biotype 2 in the USA.

A polyphasic characterization of atypical isolates of Yersinia ruckeri (causative agent of enteric redmouth disease in trout) obtained from hatchery-reared brown trout Salmo trutta in South Carolina was performed. The Y. ruckeri isolates were biochemically and genetically distinct from reference cultures, including the type strain, but were unequivocally ascribed to the species Y. ruckeri, based on API 20E, VITEK, fatty acid methyl ester profiles, and 16S rRNA gene sequencing analysis. These isolates were nonmotile and unable to hydrolyze Tween 20/80 and were therefore classified as Y. ruckeri biotype 2. Genetic fingerprint typing of the isolates via enterobacterial repetitive intergenic consensus (amplified by polymerase chain reaction) and fragment length polymorphism showed biotype 2 as a homogeneous group distinguishable from other Y. ruckeri isolates. This is the first report of Y. ruckeri biotype 2 in the USA.

PCR method for detection of largemouth bass virus.

A polymerase chain reaction (PCR) method was developed for largemouth bass virus (LMBV). This iridovirus can cause a lethal disease of largemouth bass Micropterus salmoides, but also subclinically infects largemouth bass and other species of fishes. Oligonucleotide primers were designed to specifically amplify the major capsid protein gene of LMBV. The protocol for sample processing and PCR provided a method that was more sensitive than cell culture for detection of LMBV in fish. The specific amplification of LMBV also provided an improved method for confirming the identity of cell-culture isolates presumptively identified as LMBV.

First isolation of largemouth bass virus.

The first reported fish kill caused by largemouth bass virus (LMBV) occurred in 1995 in Santee-Cooper Reservoir, South Carolina, USA. Subsequently, this iridovirus has been implicated in additional fish kills and has also been found in clinically healthy fish in numerous locations in the southeastern USA. We compared the virus from Santee-Cooper Reservoir with a virus isolated in 1991 from large-mouth bass, Micropterus salmoides, from Lake Weir, Florida. Restriction fragment length polymorphisms and the DNA sequence of a portion of the major capsid protein gene were identical for the South Carolina and Florida isolates. These results establish that LMBV was first found in Florida, rather than South Carolina. We propose that the name largemouth bass virus continue to be used for this virus, rather than alternative names based on geographical origin.

Reproductive characteristics of male mosquitofish (Gambusia affinis) inhabiting a small southeastern U.S. river receiving treated domestic sewage effluent.

A population of western mosquitofish (Gambusia affinis) living below a wastewater treatment plant in the vicinity of Birmingham (AL, USA) was studied for evidence of exposure to estrogens. Mosquitofish are sexually dimorphic live-bearing fish. Males have an elongated and modified anal fin, called a gonopodium, used in mating. It has been hypothesized that exposure to estrogens and/or anti-androgens in treated wastewater might inhibit the androgen-dependent development of the gonopodium. The population in this study showed no evidence of having been exposed to endocrine-disrupting chemicals. The difference in adjusted (for overall fish size via analysis of covariance) mean gonopodium length between effluent-exposed and control populations was not significant. No detectable levels of vitellogenin were observed in the blood of any of the male mosquitofish, either from the effluent-exposed or the control population. Testes and livers were weighed and examined histologically. The fish exposed to treated wastewater effluent had significantly larger adjusted mean weights for both testis and liver, but no histological changes indicating exposure to estrogens were observed.

Effects of nonylphenol on the gonadal differentiation of the hermaphroditic fish, Rivulus marmoratus.

Nonylphenol (NP) is an estrogenic degradation product of alkylphenol polyethoxylate surfactants. In this study, the effects of NP on gonadal differentiation and development in Rivulus marmoratus (Osteichthyes, Cyprinodontiformes), a self-fertilizing, hermaphroditic species, were examined. Starting at hatching, fish were exposed to 150 or 300 microg 1(-1) NP (nominal concentrations) in a static system with daily renewal. The measured concentration of NP in the test water decreased rapidly; half-life was 8.0 h. After 60 d of exposure to NP, fish were kept in uncontaminated water for 20 d and were then preserved for histological examination. No fish exposed to 300 microg l(-1) NP (N=8) and only two of nine fish exposed to 150 microg l(-1) NP developed testicular tissue, compared with nine of 13 water-control fish and five of nine solvent-control fish. Oogenesis was also significantly inhibited by NP. None of the fish exposed to 300 microg l(-1) and only two of nine fish exposed to 150 microg l(-1) NP had vitellogenic oocytes, compared with seven of 11 water-control fish (not including males) and six of nine solvent-control fish. Dysplasia of the gonadal lumen also occurred in fish exposed to 300 microg l(-1) NP. These changes, including testicular agenesis, have not been previously reported in fish exposed to NP.