Functional investigation of a QTL affecting resistance to Haemonchus contortus in sheep

This study reports a functional characterization of a limited segment (QTL) of sheep chromosome 12 associated with resistance to the abomasal nematode Haemonchus contortus. The first objective was to validate the identified QTL through the comparison of genetically susceptible (N) and resistant (R) sheep produced from Martinik × Romane back-cross sheep. The R and N genotype groups were then experimentally infected with 10 000 H. contortus larvae and measured for FEC (every three days from 18 to 30 days post-challenge), haematocrit, worm burden and fertility. Significant differences in FEC and haematocrit drop were found between R and N sheep. In addition, the female worms recovered from R sheep were less fecund. The second step of the characterization was to investigate functional mechanisms associated with the QTL, thanks to a gene expression analysis performed on the abomasal mucosa and the abomasal lymph node. The gene expression level of a candidate gene lying within the QTL region (PAPP-A2) was measured. In addition, putative interactions between the chromosome segment under study and the top ten differentially expressed genes between resistant MBB and susceptible RMN sheep highlighted in a previous microarray experiment were investigated. We found an induction of Th-2 related cytokine genes expression in the abomasal mucosa of R sheep. Down-regulation of the PAPP-A2 gene expression was observed between naïve and challenged sheep although no differential expression was recorded between challenged R and N sheep. The genotyping of this limited region should contribute to the ability to predict the intrinsic resistance level of sheep.     

Arctic charr in Britain and Ireland – 15 species or one?

Abstract –  From both a modern and a historical perspective there is little doubt that the Arctic charr, Salvelinus alpinus , in Britain and Ireland (as well as elsewhere) is a 'difficult' species. Historically 15 separate species have been recognised from populations in Britain and Ireland and there have been recent attempts to reassert these specific names. Here we review the evidence for the status of these 'species'. We conclude that the evidence for these 15 being afforded full species status is poor. However, both historical and contemporary data show that Salvelinus alpinus (Linnaeus 1758) in Britain and Ireland exhibits levels of variability in form that is much greater than in other species. We argue that a comprehensive genotypic and phenotypic survey of charr populations in Britain and Ireland is required to determine the full extent of variability and the status of populations with a view of providing suitable protection.     

Identification of Mycobacterium spp. isolated from snakehead, Channa striata (Fowler), and Siamese fighting fish, Betta splendens (Regan), using polymerase chain reaction–reverse cross blot hybridization (PCR–RCBH)

A variety of methods have been used to identify Mycobacterium spp. isolated from snakehead and Siamese fighting fish, including biochemistry, mycolic acid profiles and antibody-based methods. However, these methods are unable to differentiate between different species of Mycobacterium . Polymerase chain reaction (PCR) followed by reverse cross blot hybridization (RCBH) was adapted in this study to speciate aquatic mycobacteria. The method was highly specific for Mycobacterium spp. and identified the bacteria to species level with a detection limit of 100 fg DNA, equivalent to 20 mycobacteria. Twenty-nine isolates previously collected and cultured from Siamese fighting fish (10 isolates) and snakehead (19 isolates) during outbreaks of mycobacteriosis were analysed using PCR–RCBH. Six of the Siamese fighting fish isolates and nine of the snakehead isolates were identified as Mycobacterium fortuitum , while the remainder were classified as M. marinum . Notably, two isolates recovered from snakehead and Siamese fighting fish, previously identified as M. poriferae and M. piscicida , respectively, were confirmed to be M. fortuitum .     

Volume and Value of Marine Ornamentals Collected in Florida, 1990–98

The marine life fishing industry in Florida is defined by the state as the non-lethal harvest of marine plants, finfish, and invertebrates that are sold live for commercial purposes (primarily into the saltwater aquarium industry). Approximately 330 different species of finfish and invertebrates are harvested by marine life collectors in Florida, including 180 species of finfish and 150 species of invertebrates and plants. In 1998, the total dockside value of these species was approximately US$1.9 million. The industry is highly regulated via limits on gear, handling methods, harvest sizes, and trip/bag/possession limits. Entry into the industry has been recently curtailed by the implementation of a moratorium on marine life endorsements. The total number of licensed harvesters exceeded 700 in 1998. Approximately 70 wholesales buyers also participated in the market during 1998. The majority of the dockside value is generated in south Florida, with the majority of the remaining value accruing from the region including and north of Tampa Bay. Over 80% of the total value associated with both finfish and invertebrates is generated by only 10 species within each group. Dockside prices vary considerably among the various species landed, with those landed in greater volumes exhibiting the lower prices.     

A comparison of the antigenicity of the extracellular products and whole-cell sonicates from Mycobacterium spp. in rabbits, mice and fish by immunoblotting and enzyme-linked immunosorbent assay

The antigenicity of extracellular products (ECPs) derived from Mycobacterium spp. isolated from snakehead, Channa striata (Bloch), and Siamese fighting fish, Betta splendens (Regan), were examined by immunoblotting and enzyme-linked immunosorbent assay (ELISA) using sera collected from immunized rabbits, mice and fish (rainbow trout). All three species responded to a 65-kDa protein present in both the ECPs and whole cell sonicates (WCSs) from a variety of Mycobacterium spp. Cross-reactivity of anti- M. tuberculosis and anti-human heat-shock protein monoclonal antibodies (MAbs), and the presence of fibronectin binding proteins secreted into ECPs of mycobacteria were also examined. The MAbs against human 60-kDa heat-shock protein cross- reacted with the band at 65 kDa in the ECPs of TB1 (isolated from snakehead fish) and the type strain M. marinum, while the anti- M. tuberculosis MAb F29–47 elicited a strong reaction with a band at 21 kDa with most of the ECPs from mycobacterial strains examined. The major fibronectin-binding proteins were located between 21 and 25 kDa. The 65-kDa protein from ECPs of Mycobacterium spp. proved strongly immunogenic to rabbits, mice and fish. Rabbit antiserum against the 65-kDa protein from strain TB267 reacted with many non- Mycobacterium WCSs, and therefore, the 65-kDa protein from Mycobacterium spp. is believed to be a common protein found in many fish bacterial pathogens.     

Ecology of larval blue sucker (Cycleptus elongatus) in the Mississippi River

Abstract –  Ecology of larval blue sucker, Cycleptus elongatus , a North American catostomid that has declined throughout its range, is poorly known because larvae have rarely been sampled. A total of 316 young blue suckers (16.0–39.0 mm total length) was captured over 3 years at 14 off-channel sites in Pool 25, Mississippi River. Blue suckers demonstrated distinct temporal and spatial abundance patterns. Blue suckers were only captured in late May/early June, and were more abundant during a year of little flooding. Abundance was correlated with the distance a site was from the main channel, being highest in island borders and island sloughs in proximity to the channel. Fish guts contained a relatively high biomass of chironomids and zooplankton, suggesting islands were providing suitable feeding opportunities. Because of their proximity to flows, slack waters provided by islands were accessible to blue sucker larvae dispersing from channel spawning areas and facilitated their development into rheophilic juveniles.     

Experimental amoebic gill disease of Atlantic salmon, Salmo salar L.: further evidence for the primary pathogenic role of Neoparamoeba sp. (Page, 1987)

Amoebic gill disease (AGD) has been attributed to infection by Neoparamoeba sp. The causal mechanisms for AGD lesion development and the primary pathogenic role of Neoparamoeba sp. require elucidation. Three groups of Atlantic salmon were exposed to viable gill isolated amoebae, to sonicated amoebae, or to sea water containing viable amoebae without direct contact with gill epithelia. Fish were removed 8 days post-exposure and the gills assessed histologically for AGD. AGD occurred only when fish were exposed to viable trophozoites. Consequently, in an accompanying experiment, infection was evaluated histologically at 12, 24 and 48 h post-exposure in three groups of salmon, one group being mechanically injured 12 h prior to exposure. A progressive host response and significant increase (P < 0.001) in the numbers of attached amoebae was apparent over the 48-h duration in undamaged hemibranchs in both treatment groups. There were no significant differences to mucous cell populations. Attachment of Neoparamoeba sp. to damaged gill filaments was significantly reduced (P < 0.05) by 48 h post-exposure. These data further confirm and describe the primary pathogenic role of Neoparamoeba sp. and the early host response in AGD. Preliminary evidence suggests that lesions resulting from physical gill damage are not preferentially colonized by Neoparamoeba sp.