Transcriptome analyses of amoebic gill disease-affected atlantic salmon (Salmo salar) tissues reveal localized host gene suppression

The transcriptome response of Atlantic salmon (Salmo salar) displaying advanced stages of amoebic gill disease (AGD) was investigated. Naïve smolt were challenged with AGD for 19 days, at which time all fish were euthanized and their severity of infection quantified through histopathological scoring. Gene expression profiles were compared between heavily infected and naïve individuals using a 17 K Atlantic salmon cDNA microarray with real-time quantitative RT-PCR (qPCR) verification. Expression profiles were examined in the gill, anterior kidney, and liver. Twenty-seven transcripts were significantly differentially expressed within the gill; 20 of these transcripts were down-regulated in the AGD-affected individuals compared with naïve individuals. In contrast, only nine transcripts were significantly differentially expressed within the anterior kidney and five within the liver. Again the majority of these transcripts were down-regulated within the diseased individuals. A down-regulation of transcripts involved in apoptosis (procathepsin L, cathepsin H precursor, and cystatin B) was observed in AGD-affected Atlantic salmon. Four transcripts encoding genes with antioxidant properties also were down-regulated in AGD-affected gill tissue according to qPCR analysis. The most up-regulated transcript within the gill was an unknown expressed sequence tag (EST) whose expression was 218-fold (± SE 66) higher within the AGD affected gill tissue. Our results suggest that Atlantic salmon experiencing advanced stages of AGD demonstrate general down-regulation of gene expression, which is most pronounced within the gill. We propose that this general gene suppression is parasite-mediated, thus allowing the parasite to withstand or ameliorate the host response. © 2008 Springer Science+Business Media, LLC.

Resistance to amoebic gill disease (AGD) is characterised by the transcriptional dysregulation of immune and cell cycle pathways

Amoebic gill disease (AGD) is a parasite-mediated proliferative gill disease capable of affecting a range of teleost hosts. While a moderate heritability for AGD resistance in Atlantic salmon has been reported previously, the mechanisms by which individuals resist the proliferative effects remain poorly understood. To gain more knowledge of this commercially important trait, we compared gill transcriptomes of two groups of Atlantic salmon, one designated putatively resistant, and one designated putatively susceptible to AGD. Utilising a 17k Atlantic salmon cDNA microarray we identified 196 transcripts that were differentially expressed between the two groups. Expression of 11 transcripts were further examined with real-time quantitative RT-PCR (qPCR) in the AGD-resistant and AGD-susceptible animals, as well as non-infected naïve fish. Gene expression determined by qPCR was in strong agreement with the microarray analysis. A large number of differentially expressed genes were involved in immune and cell cycle responses. Resistant individuals displayed significantly higher expression of genes involved in adaptive immunity and negative regulation of the cell cycle. In contrast, AGD-susceptible individuals showed higher expression of acute phase proteins and positive regulators of the cell cycle. Combined with the gill histopathology, our results suggest AGD resistance is acquired rather than innately present, and that this resistance is for the most part associated with the dysregulation of immune and cell cycle pathways. © 2008 Elsevier Ltd. All rights reserved.

Analysis, characterisation and expression of gill-expressed carbonic anhydrase genes in the freshwater crayfish Cherax quadricarinatus

Changes in water quality parameters such as pH and salinity can have a significant effect on productivity of aquaculture species. Similarly, relative osmotic pressure influences various physiological processes and regulates expression of a number of osmoregulatory genes. Among those, carbonic anhydrase (CA) plays a key role in systemic acid–base balance and ion regulation. Redclaw crayfish (Cherax quadricarinatus) are unique in their ability to thrive in environments with naturally varied pH levels, suggesting unique adaptation to pH stress. To date, however, no studies have focused on identification and characterisation of CA or other osmoregulatory genes in C. quadricarinatus. Here, we analysed the redclaw gill transcriptome and characterized CA genes along with a number of other key osmoregulatory genes that were identified in the transcriptome. We also examined patterns of gene expression of these CA genes when exposed to three pH treatments. In total, 72,382,710 paired end Illumina reads were assembled into 36,128 contigs with an average length of 800 bp. Approximately 37% of contigs received significant BLAST hits and 22% were assigned gene ontology terms. Three full length CA isoforms; cytoplasmic CA (ChqCAc), glycosyl-phosphatidylinositol-linked CA (ChqCAg), and β-CA (ChqCA-beta) as well as two partial CA gene sequences were identified. Both partial CA genes showed high similarity to ChqCAg and appeared to be duplicated from the ChqCAg. Full length coding sequences of Na+/K+-ATPase, V-type H+-ATPase, sarcoplasmic Ca+-ATPase, arginine kinase, calreticulin and Cl− channel protein 2 were also identified. Only the ChqCAc gene showed significant differences in expression across the three pH treatments. These data provide valuable information on the gill expressed CA genes and their expression patterns in freshwater crayfish. Overall our data suggest an important role for the ChqCAc gene in response to changes in pH and in systemic acid–base balance in freshwater crayfish.

Comparative analysis of gill transcriptomes of two freshwater crayfish, Cherax cainii and C. destructor

We undertook deep sequencing of gill transcriptomes from two freshwater crayfish, Cherax cainii and Cherax destructor, in order to generate genomic resources for future genomics research. Over 83 and 100 million high quality (quality score (Q) ≥ 30) paired-end Illumina reads (150 bp) were assembled into 147,101 and 136,622 contigs in C. cainii and C. destructor, respectively. A total of 24,630 and 23,623 contigs received significant BLASTx hits and allowed the identification of multiple gill expressed candidate genes associated with pH and salinity balance. These functionally annotated transcripts will provide a resource to facilitate comparative genomic research in the genus Cherax, and in particular allow insights into respiratory and osmoregulatory physiology of this group of animals.

Transcriptome analysis and characterisation of gill-expressed carbonic anhydrase and other key osmoregulatory genes in freshwater crayfish Cherax quadricarinatus

The pH and salinity balance mechanisms of crayfish are controlled by a set of transport-related genes. We identified a set of the genes from the gill transcriptome from a freshwater crayfish Cherax quadricarinatus using the Illumina NGS-sequencing technology. We identified and characterized carbonic anhydrase (CA) genes and some other key genes involved in systematic acid-base balance and osmotic/ionic regulation. We also examined expression patterns of some of these genes across different sublethal pH levels [1]. A total of 72,382,710 paired-end Illumina reads were assembled into 36,128 contigs with an average length of 800 bp. About 37% of the contigs received significant BLAST hits and 22% were assigned gene ontology terms. These data will assist in further physiological-genomic studies in crayfish.

Effects of temperature on complexes I and II mediated respiration, ROS generation and oxidative stress status in isolated gill mitochondria of the mud crab Scylla serrata

Effects of fluctuations in habitat temperature (18-30 degrees) on mitochondrial respiratory behavior and oxidative metabolic responses in the euryhaline ectotherm Scylla serrate are not fully understood. In the present study, effects of different temperatures ranging from 12 to 40 degrees C on glutamate and succinate mediated mitochondrial respiration, respiratory control ratio (RCR), ATP generation rate, ratio for the utilization of phosphate molecules per atomic oxygen consumption (P/O), levels of lipid peroxidation and H2O2 in isolated gill mitochondria of S. serrata are reported. The pattern of variation in the studied parameters was similar for the two substrates at different temperatures. The values recorded for RCR ( >= 3) and P/O ratio (1.4-2.7) at the temperature range of 15-25 degrees C were within the normal range reported for other animals (3-10 for RCR and 1.5-3 for P/O). Values for P/O ratio, ATP generation rate and RCR were highest at 18 degrees C when compared to the other assay temperatures. However, at low and high extreme temperatures, i.e. at 12 and 40 degrees C, states III and IV respiration rates were not clearly distinguishable from each other indicating that mitochondria were completely uncoupled. Positive correlations were noticed between temperature and the levels of both lipid peroxidation and H2O2. It is inferred that fluctuations on either side of ambient habitat temperature may adversely influence mitochondria respiration and oxidative metabolism in S. serrata. The results provide baseline data to understand the impacts of acute changes in temperature on ectotherms inhabiting estuarine or marine environments. (C) 2014 Elsevier Ltd. All rights reserved.

A study of the length at maturity and hooks/gill net mesh selection in Clarias lazera from Lake Chad

The minimum length at first maturity of Clarias lazera was found to be 24 cm (4.8%) for females and 20 cm (1.8%) for males. Fifty percent maturity was attained at length of 28 cm to 30 cm for both sexes; there being little difference among the sexes at this level of maturity. The modal retention lengths for gill nets were: 13 cm for 25.5 mm mesh; 18 cm for 32 mm mesh; 28 cm for 57 mm mesh; and 38 cm for 76 mm mesh. Modal lengths of Clarias lazera caught by various hooks sizes were No. 10 (28 cm); No. 11 (33 cm); Nos. 15 and 16 (28 cm). It is recommended that to protect the clarias fishery in Lake Chad, the use of gill nets of less than 57 mm mesh size and fishing hook No. 16 (and smaller sizes) which caught 43.94% of immature fishes should be discouraged

An evaluation of geographic and annual variation in morphometric characters and gill-raker counts of yellowfin tuna, Thunnus albacares, from the Pacific Ocean

ENGLISH: Samples of yellowfin tuna, Thunnus albacares, collected from five areas of the Pacific Ocean (Mexico, Ecuador, Australia, Japan, and Hawaii) between January and May of 1988 and 1990 were examined for spatiotemporal variation in morphometric characters and gill-raker counts. 'Iwo-factor analysis of variance, with area and year treated as grouping factors, indicated a significant difference in the means of the total gill-raker counts among fish from different areas, but no significant difference between fish caught in different years. The morphometric data were adjusted by allometric formulae to remove size effects. The correct classification rates for the five groups, using discriminant function analysis, based on adjusted morphometric characters, were 77.60/0 for the samples from 1988 and 74.40/0 for those from 1990. These are 72.00/0 and 68.00/0 (Cohen's kappa statistic) better than would have occurred chance. The pattern of geographic variability, however, is unstable for these two years, thus requiring separate discriminant functions for each year. Although there is annual variability in the morphometric characters, these results demonstrate that the stocks examined are morphometrically distinguishable and that their phenetic relationships reflect their geographic origin. SPANISH: Se examinaron muestras de atún aleta amarilla, Thunnus albacares, tomadas de cinco áreas del Océano Pacífico (México, Ecuador, Australia, Japón, y Hawaii) entre enero y mayo de 1988 y 1990, para descubrir variaciones espaciotemporales en las características morfométricas y los conteos de branquiespinas. Un análisis de varianza de dos factores, con área y año como factores de agrupación, indicó una diferencia significativa en los promedios de los conteos de branquiespinas totales entre peces de distintas áreas, pero ninguna entre peces capturados en distintos años. Se ajustaron los datos morfométricos con fórmulas alométricas para eliminar los efectos de la talla del pez. En un análisis de función discriminante, las tasas de clasificación correcta de los cinco grupos, basadas en características morfométricas ajustadas, fueron 77.60/0 para las muestras de 1988 y 74.40/0 para aquellas de 1990. Estas cifras son 72.00/0 y 68.00/0 (estadístico de kappa de Cohen) mejores de lo que se hubiera obtenido al azar. Sin embargo, la variabilidad geográfica es inestable en estos dos años, requiriendo por lo tanto funciones discriminantes separadas para cada año. Aunque existe variabilidad anual en las características morfométricas, estos resultados demuestran que los stocks examinados son morfamétricamente distinguibles, y que su relación fenética refleja su origen geográfico. (PDF contains 31 pages.)

The effect of long term exploitation by the gill net fishery on the multispecies fish stocks in Kainji Lake, Nigeria, 1969-1997

The findings are presented of an assessment made of the gillnet fishery in Kainji Lake, Nigeria from 1969 to the present, on the basis of data sets from commercial and experimental gillnet fishing, with the purpose to detect trends in some key fishery monitoring indicators. During this period, there has been an increase in the number of small meshed nets in the fishery resulting in a shift in the mode to lower mesh sizes; consequently, the average mesh size declined gradually in the fishery. This trend is found to be directly correlated with the decline in the CPUE and mean weight of the fish species. It is argued that the observed trend in CPUE and mean weight is forcing the fishermen to switch effort to gears such as traps which have very small meshes and can indescriminately take all sizes of the fish. It is shown that the catch composition by weight of Citharinus citharus, Lates niloticus and tilapias declined in the gillnet fishery in the late 70's and early 80's. Recent data, from 1994 to 1996, however indicates that C. citharus is recovering, but with declining mean weight. This suggests that the exploitation pattern is shifting to the smaller fish through the use of small meshed nets. In general, however, there has not been drastic changes in species bio-diversity in the Lake as a result of predatory effect and ecosystem overfishing as has happened in other great African Lakes. The species composition since lake formation continued to be dominated by fewer that 20 species. The potential yield for the lake has been estimated to be 32,166 tonnes (excluding clupeids) and the required optimum fishing effort to be 1,814 fishing canoes. In view of the relative stability of the species diversity in the lake and the current fish production level, it is proposed here that this MSY be adopted for all species. This would be achieved with the current effort level in the lake assuming that the efficiency of the fishermen and their gears do not improve. It should be reviewed after 10 or more years of catch and effort data collection. (PDF contains 65 pages)

Some statistical aspects of the long-term gill net monitoring programme for pike Esox lucius in Windermere (English Lake District)

For more than 55 years, data have been collected on the population of pike Esox lucius in Windermere, first by the Freshwater Biological Association (FBA) and, since 1989, by the Institute of Freshwater Ecology (IFE) of the NERC Centre for Ecology and Hydrology. The aim of this article is to explore some methodological and statistical issues associated with the precision of pike gill net catches and catch-per-unit-effort (CPUE) data, further to those examined by Bagenal (1972) and especially in the light of the current deployment within the Windermere long-term sampling programme. Specifically, consideration is given to the precision of catch estimates from gill netting, including the effects of sampling different locations, the effectiveness of sampling for distinguishing between years, and the effects of changing fishing effort.

Mechanisms of Toxicity of Ag Nanoparticles in Comparison to Bulk and Ionic Ag on Mussel Hemocytes and Gill Cells

Silver nanoparticles (Ag NPs) are increasingly used in many products and are expected to end up in the aquatic environment. Mussels have been proposed as marine model species to evaluate NP toxicity in vitro. The objective of this work was to assess the mechanisms of toxicity of Ag NPs on mussel hemocytes and gill cells, in comparison to ionic and bulk Ag. Firstly, cytotoxicity of commercial and maltose stabilized Ag NPs was screened in parallel with the ionic and bulk forms at a wide range of concentrations in isolated mussel cells using cell viability assays. Toxicity of maltose alone was also tested. LC50 values were calculated and the most toxic Ag NPs tested were selected for a second step where sublethal concentrations of each Ag form were tested using a wide array of mechanistic tests in both cell types. Maltose-stabilized Ag NPs showed size-dependent cytotoxicity, smaller (20 nm) NPs being more toxic than larger (40 and 100 nm) NPs. Maltose alone provoked minor effects on cell viability. Ionic Ag was the most cytotoxic Ag form tested whereas bulk Ag showed similar cytotoxicity to the commercial Ag NPs. Main mechanisms of action of Ag NPs involved oxidative stress and genotoxicity in the two cell types, activation of lysosomal AcP activity, disruption of actin cytoskeleton and stimulation of phagocytosis in hemocytes and increase of MXR transport activity and inhibition of Na-K-ATPase in gill cells. Similar effects were observed after exposure to ionic and bulk Ag in the two cell types, although generally effects were more marked for the ionic form. In conclusion, results suggest that most observed responses were due at least in part to dissolved Ag.

The effect of mesh size on the fishing efficiency of sardine gill nets

The study deals with the effect of mesh size on the fishing power of gill nets. The authors have shown that there can be substantial difference in the out puts of sardine gill nets, of identical design and rigging, but of different mesh sizes, operated under the same conditions.