Detection and localization of oxidized proteins in muscle cells by fluorescence microscopy

In meat, no detailed studies on the intracellular distribution of oxidized proteins during oxidative stress have been performed, to our knowledge. Therefore, we used fluorescence microscopy to detect and locate protein carbonyls, oxidation products of basic amino acids, generated in bovine M. Rectus abdominis during either exposition to a chemical free radical generating system, or refrigerated storage, or cooking. The technique consisted of an immunohistochemical detection of carbonyls by reaction with the specific probe DNPH (2,4-dinitrophenylhydrazine) followed by the sequential addition of a first antibody against DNPH-carbonylated proteins and a CY3-labeled secondary antibody. The fluorescence of the CY3 probe increased regularly with level of free radical generating system and storage time. Moreover, an important heterogeneity of carbonyl distribution was observed, with a higher oxidation level at the periphery than inside the muscle cells. Cooking induced fluorescence increase only at the periphery of cells. Specific coloration of collagen by Sirius red showed that collagen was not involved in fluorescence. We can deduce that accumulation of oxidized proteins observed in the cell periphery was linked to membrane protein oxidation and not to connective tissue oxidation. Biochemical assays were performed in parallel on membrane and myofibrillar proteins to provide complementary quantitative data on level of oxidized proteins.     

Thermal biology of Tuta absoluta: demographic parameters and facultative diapause

Journal of Pest Science - The South American tomato pinworm, Tuta absoluta, (SATP) is now a devastating pest worldwide of crops in the family Solanaceae. Most prior studies of SATP’s thermal...     

Modeling the egg and larval transport pathways of the Antarctic toothfish (Dissostichus mawsoni) in the East Antarctic region: New insights into successful transport connections

The Antarctic toothfish (Dissostichus mawsoni) is an important fishery species widely distributed in the Southern Ocean, especially in areas covered by sea ice. Understanding fish distributions and life cycles, including the transport and survival of eggs and larvae, is essential for the assessment and sustainable management of the fishery. However, owing to difficulties with in situ winter observations, information on the early life stages of D. mawsoni is lacking. Here, we investigated the transport pathways of fish eggs and larvae through a particle tracking study, using satellite-derived ocean surface velocities in the East Antarctic region, which includes important D. mawsoni habitats and exploratory fisheries. Our results indicate that particles released from continental slopes are more likely to be successfully transported to suitable settlement grounds than those released from the BANZARE Bank (the southern region of the Kerguelen Plateau), which is situated further north and has been hypothesized to be a potential spawning ground for D. mawsoni. This study demonstrates successful source–settlement connections in relation to ocean recirculation and suggests important settlement regions for D. mawsoni larvae in the East Antarctic region.     

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.     

Coastal and interior Douglas-fir provenances differ in growth performance and response to drought episodes at adult age

Context

Since the 2003 drought and heat wave, Douglas-fir dieback has been reported in France in trees older than 30 years. Consequently, it is questioned whether selected Douglas-fir provenances are suited to the frequent and severe drought events which are forecast due to climate change.

Aims

Our objective was to contribute to the screening of variability in productivity and growth response to soil-water deficit of mature trees from provenances not currently used for plantation in France.

Methods

We sampled 22 provenances, including coastal and interior Douglas-fir, covering a wide part of its natural distribution, from Oregon to California for coastal provenances and from British Columbia to New Mexico for interior provenances. These provenances were planted at the mid 1970s in two provenance trials located in the south-west area of France. Variability of productivity, of wood density, and of radial growth in response to drought episodes among provenances was quantified and related to soil-water deficit computed by daily water balance calculations.

Results

Whatever the provenance, annual radial growth is highly dependent on local soil-water deficit (Felines R ² = 0.57, Le Treps R² = 0.49). Radial growth and wood properties exhibit large differences between provenances at 30 years old. Variability between provenances for all wood characteristics studied is mainly structured geographically. Coastal provenances perform best for productivity at 30 years old (619 cm²?±?59), and exhibit a small growth reduction in 2004, the second successive year of drought (?10.7 %?±?3.8). Surprisingly, the southern interior provenances from the driest environments in the natural range show a large growth reduction in 2004 (?30.5 %?±?5.2).

Conclusions

The provenances tested exhibited significant differences in growth performance and growth reduction induced by the soil-water deficit. The approach coupling retrospective analysis of radial growth on mature trees and water balance modelling is relevant for evaluating provenance adaptation to more frequent or severe drought episodes.     

The effect of different HUFA enrichment emulsions on the nutritional value of rotifers (Brachionus plicatilis) fed to larval haddock (Melanogrammus aeglefinus).

A feeding study was conducted in the winter 2001 to determine the effects of feeding rotifers (Brachionus plicatilis) enriched with various levels of essential fatty acids on the growth and survival of haddock larvae (Melanogrammus aeglefinus). Rotifer enrichment treatments were: 1) mixed algae, 2) high DHA (docosahexaenoic acid, 22:6n-3), 3) high DHA and EPA (eicosapentaenoic acid, 20:5n-3), and 4) DHA, EPA, and AA (arachidonic acid, 20:4n-6). Larvae were fed rotifers enriched with the different treatments from days 1 to 16 post-hatch. From day 17 until 25 all treatment groups were fed rotifers reared on mixed algae and then weaned onto the International Council for Exploration of the SEA (ICES) Standard Reference Weaning diet (http://allserv.rug.ac.t/aquaculture/rend/rend.htm) over a five day period. The experiment was terminated on day 41 post-hatch. The enrichment treatments affected the fatty acid composition of the rotifers and correlated with the accumulation of these fatty acids in the haddock larvae. However, no significant differences in larval growth or survival to 40 days post hatch were detected, suggesting that all treatments provided the minimal essential fatty acid requirements for haddock.     

In vitro modulation of reactive oxygen and nitrogen intermediate (ROI/RNI) production in Crassostrea gigas hemocytes

Bivalve hemocyte competence has been measured by quantifying functional characteristics, including reactive oxygen intermediate (ROI) production after activation with zymosan or phorbol myristate acetate (PMA). However, untreated oyster hemocytes also produce ROI and RNI (reactive nitrogen intermediates) after bleeding even if not stimulated by zymosan or PMA. Extensive investigation of this parameter by flow cytometry showed that, in vitro, ROI/RNI production by untreated hemocytes maintained in seawater appeared to be independent of both bacterial burden in the serum and non-self particle phagocytosis. ROI/RNI production in granulocytes was higher than in hyalinocytes and could be intensified when activated by zymosan but not by PMA. Both cell types used NADPH-oxidase- and NO-synthase-like pathways to produce these molecules; the NO-synthase pathway seemed relatively more dominant in hyalinocytes and NADPH-oxidase appeared more effective in granulocytes. These results provide new insights for interpreting the modulation of ROI/RNI production by untreated hemocytes shown by other studies, relative to environmental conditions or physiological status of the oysters.     

Recent developments and improvements in ornamental fish packaging systems for air transport

Current ornamental fish packaging systems are characterized by very high fish loading densities and high metabolic wastes in the transport water after shipment. They focus mainly on management of the quality of transport water. Recent studies using the guppy as a model fish showed that post‐shipment mortality could be reduced through enhancement of the stress resistance of the fish, and hence emphases should also be placed on the preparation of the fish for transport and recovery of the fish after shipment. Farmers can contribute significantly by applying nutritional prophylaxis before harvesting. Exporters may use the salinity stress test to identify fish lots of good quality for transport, apply health prophylaxis to eradicate parasites and optimize other techniques such as starvation of the fish or addition of salt to the transport water to enhance the stress resistance of the fish. Importers may adopt proper acclimation procedure and allow fish to recover in low salinity water to reduce post‐shipment mortality. As the main bulk of post‐shipment mortality is stress‐mediated and occurs during the 1‐week recovery period, the industry should consider revising the basis of the current warranty system for their customers, from death on arrival to cumulative mortality at 7 days post shipment (or death after 7 days, DA7), in order to cut down fish losses after shipment.     

Automation in stock-culture maintenance and juvenile separation of the mysid Mysidopsis bahia (molenock)

A batch system with subgravel filter for stock-culture maintenance of Mysidopsis bahia in artificial sea water is described. The feeding of mysids on Artemia is optimized through the cyclic pumping of brine shrimp instar I nauplii from a refrigerator to the stock-cultures. An easily constructed incubator-separator apparatus is described for the standardized harvesting of mysid juveniles to be used as test organisms.     

Link between Domoic Acid Production and Cell Physiology after Exchange of Bacterial Communities between Toxic Pseudo-nitzschia multiseries and Non-Toxic Pseudo-nitzschia delicatissima

Bacteria are known to influence domoic acid (DA) production by Pseudo-nitzschia spp., but the link between DA production and physiology of diatoms requires more investigation. We compared a toxic P. multiseries to a non-toxic P. delicatissima, investigating links between DA production, physiological parameters, and co-occurring bacteria. Bacterial communities in cultures of both species were reduced by antibiotic treatment, and each of the diatoms was inoculated with the bacterial community of the other species. The physiology of P. delicatissima was minimally affected by the absence of bacteria or the presence of alien bacteria, and no DA was detected. P. multiseries grew faster without bacteria, did not produce a significant amount of DA, and exhibited physiological characteristics of healthy cells. When grown with alien bacteria, P. multiseries did not grow and produced more DA; the physiology of these cells was affected, with decreases in chlorophyll content and photosynthetic efficiency, an increase in esterase activity, and almost 50% mortality of the cells. The alien bacterial community had morphological and cellular characteristics very different from the original bacteria, and the number of free-living bacteria per algal cell was much higher, suggesting the involvement of bacteria in DA production.