Temperature modulates liver lipid accumulation in Atlantic salmon (Salmo salar L.) fed low dietary levels of long‐chain n‐3 fatty acids

Atlantic salmon (Salmo salar) were fed five graded levels of eicosapentaenoic acid (EPA, 20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3), from 1.4 to 5.2% of total fatty acids (FA, 5–17 mg kg^?1 feed), and grew from ~160 g to ~3000 g, with the period from 1450 g onwards conducted both at 6 °C and at 12 °C. All fish appeared healthy, and there were no diet‐related differences in haematological or plasma parameters, as well as intestinal histological or gut microbiota analysis. Fish reared at 6 °C had higher accumulation of storage lipids in the liver compared to fish reared at 12 °C. Liver lipids also increased with decreasing dietary EPA + DHA at 6 °C, while there was no such relationship at 12 °C. Gene expression of SREBP1 and 2, LXR, FAS and CPT1 could not explain the differences in liver lipid accumulation. In liver polar lipids, DHA was found to be reduced when dietary EPA + DHA was <2.7% of FAs, while the level of EPA in the membranes was not affected. In conclusion, reducing dietary EPA + DHA from 5.2 to 1.4% of total FAs had a minor impact on fish health. Temperature was the factor that most affected the liver lipid accumulation, but there was also an interaction with dietary components.     

Partial replacement of soybean meal with Methylobacterium extorquens single‐cell protein in feeds for rainbow trout (Oncorhynchus mykiss Walbaum)

A feeding trial was conducted with juvenile rainbow trout (15–16 g initial weight) to assess the effects of including single‐cell protein (SCP) produced from Methylobacterium extorquens in trout feeds. Three isonitrogenous and isoenergetic diets were produced: a control diet and two experimental diets containing 5% or 10% bacterial protein meal replacing soybean meal. Triplicate tanks, each containing 35 fish, were fed each diet to apparent satiation in a constant‐temperature (15°C), flow‐through tank system for 12 weeks. No statistically significant differences in final fish weight or other fish growth parameters were observed. Similarly, feed efficiency parameters showed no significant differences among groups. Nutrient retention indices (protein, fat, energy) were relatively high and similar among fish in each dietary treatment group, as were whole body proximate compositions. Fish survival was high, with a small but statistically significant increase for the 10% SCP diet. Overall, results demonstrate that SCP from M. extorquens is a safe and effective alternative protein for rainbow trout diets at the low inclusion levels tested. Slightly lower weight gain in fish fed the 10% SCP diet was largely due to lower feed intake, suggesting that adding palatability‐enhancing ingredients to feeds may allow higher levels of M. extorquens SCP to be used without compromising fish growth.     

Calcium nutrition of the sugarbeet

Abstract

When sugarbeet seedlings are transferred from a complete nutrient solution to one from which Ca has been withheld, the rootlets and tops fail to develop. The same transfer at the eight‐leaf stage causes the rootlets to become stubby and swollen at the tips and blade expansion becomes modified; particularly the upper portions of the blades attaining nearly full development, which pucker and often develop a cupping or hooding effect; a unique symptom characteristic of Ca deficiency. As each new leaf develops, the blade area becomes smaller until only a black tip remains at the apex of the petiole, which is the symptom referred to as tip‐burn for this petiole and the successively . shorter petioles formed as Ca deficiency increases in severity. Strangely, these symptoms also appear during periods of rapid growth when the nutrient solution contains as much as 10 to 28 milliequivalents per liter of Ca or when soils are high in Ca. This implies that Ca absorption and possibly translocation limits the Ca supply at the growing point. Increasing Mg in the nutrient solution decreases Ca uptake and increases Ca deficiency. Potassium deficiency, unexpectedly, induces Ca deficiency apparently by decreasing the translocation of Ca to the growing point.

These phenomena suggest the hypothesis that when ion absorption takes place from the root exchange site that has the affinity for H > Ca > Mg > K > Na, then the H generated internally replaces, and the roots absorb, Na, K and Mg preferentially. Externally, Ca would be adsorbed preferentially from the nutrient solution by the exchange complex, and with the addition of Mg, it would compete for the common adsorption site of Ca and limit Ca absorption internally. Under these conditions potassium‐deficient nutrient solutions would not induce Ca deficiency by decreasing Ca absorption but rather by decreasing Ca translocation. Theoretically, Ba would replace H more readily than Ca on the exchange complex, and therefore, Ba would be adsorbed preferentially and Ca uptake would increase. This effect of Ba was verified experimentally.

Since the translocation of ⁴⁵Ca to the growing point was found to be unrestricted under Ca‐sufficient and Ca‐deficient conditions and since the formation of insoluble Ca compounds such as phosphate or oxalate did not account for the Ca deficiency at the growing point, the cause of the Ca deficiency at the growing point is most likely the higher priority of the storage root for Ca over tops when leaf blades and storage root are both expanding rapidly. However, Ca retransport from older to younger parts of the sugarbeet plant may be restricted by the formation of Ca phosphate under Ca‐deficient conditions and Ca oxalate under Ca‐sufficient conditions.

Calcium deficiency increases net photosynthesis per unit blade area initially, probably because of blade puckering, but not on a per unit chlorophyll basis.     

Developmental temperature stress and parental identity shape offspring burst swimming performance in sockeye salmon (Oncorhynchus nerka)

Abstract – The persistent effects of embryonic temperature stress and individual parentage on fry swimming performance were examined in a cross‐fertilisation experiment using sockeye salmon (Oncorhynchus nerka). A fixed‐velocity test of burst swimming was used to assess the endurance capacity and behavioural performance of individual fry from 10 offspring families incubated at 12, 14 or 16 °C to hatch and then reared through yolk absorption and exogenous feeding stages in a common posthatch environment (average 6.9 °C). Fry burst swim time (BST) was influenced by an interaction between incubation temperature and family identity. Average BST was longer for fry from the 12 °C prehatch treatment compared to 14 and 16 °C, although differences were largely attributable to temperature effects on average fry size. Behavioural observations revealed that fish incubated at 16 °C performed more poorly, having a larger proportion of individuals that required stimulation to swim, fatigued more frequently or were classified as ‘nonswimmers’. Within all three incubation temperature treatments, mean BST varied significantly among offspring families, independent of fry mass and length. An interesting relationship was observed within the 16 °C treatment, whereby families with higher survivorship were characterised with lower mean BSTs. Collectively, these findings demonstrate that exposure to high temperatures in early sockeye salmon development can result in persistent, parentally mediated effects on fry performance. As such, these results provide important insight into how elevated temperature events during egg incubation may affect early life history selection processes and survival in stages beyond when the stressor is experienced.     

Somatic cell and innate immune responses in mammary glands of lactating cows to intramammary infusion of Bifidobacterium breve at pre-drying off period

Intramammary infusion of Bifidobacterium breve (B. breve)-induced somatic cell (SC) counts, chemiluminescent response (CL), lactoferrin (LF) concentrations and mastitis-causing pathogens from quarters with subclinical mastitis were measured to evaluate innate immune response of mammary glands in dairy cows at 3 to 4 weeks before drying off. SC counts in 7 quarters of 7 control cows and 5 quarters of 6 cows with mastitis increased markedly on day 1 and SC values in control cows were significantly (P<0.05) increased and returned to pre-infusion levels on day 5 after B. breve-infusion. CL values in both groups increased markedly on day 1 and then decreased after B. breve-infusion; however, CL values in cows with mastitis did not return to normal levels on day 5 and at postpartum. The CL values were highly correlated with their SC counts in milk from both groups. LF concentrations increased toward day 3 after B. breve-infusion and were higher in cows with mastitis. B. breve-infusion eliminated 16.6% (1/6) of pathogens from 6 quarters with chronic subclinical mastitis. B. breve-induced SC responses in quarters from 3 cows with mastitis showed characteristic patterns of recovery, persistent and new infections. B. breve-induced SC counts in quarters from the cows in the pre-drying off were lower (25.7–70.6%) than those of the cows in mid-lactation. The intrinsic innate immune response in cows on pre-drying off may be decreased and appears to be insufficient to eliminate pathogens from mammary gland in the pre-drying off.     

Factors affecting the virulence of Ralstonia solanacearum and its colonization on tobacco roots

Tobacco bacterial wilt caused by Ralstonia solanacearum is a serious disease affecting tobacco cultivation in southwest China. The response surface methodology was employed to evaluate the optimal conditions of tobacco bacterial wilt, and green fluorescent protein gene (gfp) labelling was applied to monitor the location and survival dynamics of R. solanacearum (Rs::gfp) on tobacco roots and in soil under these optimal conditions. The results showed that the highest wilt incidence was 91.13%, which occurred when the population reached 6.6 × 10⁶ CFU/g soil, the temperature was 30.55 °C, and the humidity was >81.42%. The Rs::gfp densely colonized the root tips and root hairs, and cells of Rs::gfp were observed intermittently in the elongation zone or at the point of the emerging lateral roots. The Rs::gfp number in the rhizosphere soil was 10.75‐, 73.13‐ and 74.86‐times higher than that in the bulk soil at 10, 15 and 20 days after transplantation, respectively. Increased colonization by Rs::gfp was related to the population of the pathogen, the environmental temperature and the humidity in the soil. These three conditions determined whether R. solanacearum would induce tobacco wilt. This is the first study to investigate factors affecting the virulence of a tobacco wilt bacterial pathogen, which is important for conducting field diagnosis and biocontrol of tobacco bacterial wilt.     

Effects of concentrate‐to‐forage ratios and 2‐methylbutyrate supplementation on ruminal fermentation,bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers

This study evaluated the effects of dietary concentrate levels and 2‐methylbutyrate (2MB ) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD ) in steers. Eight ruminally cannulated Simmental steers (12 months of age; 389 ± 3.7 kg of body weight) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Moderate‐concentrate (400 g/kg diet [MC ]) or high‐concentrate (600 g/kg diet [HC ]) diets were fed with or without 2MB (0 g/day [2MB ?] or 15.0 g/day [2MB +]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate‐to‐propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH ₃‐N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus , Ruminococcus flavefaciens , Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α‐amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet × 2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.     

Comparative activity of Choristoneura fumiferana nucleopolyhedrovirus propagated in different hosts

The biological activity of the Ireland strain of Choristoneura fumiferana (Clem) nucleopolyhedrovirus (CfMNPV) propagated in different hosts was determined to provide the basis upon which genetically modified CfMNPV, or other naturally occurring isolates, should be compared. Occlusion bodies (OB) derived from CF-203 cells were significantly larger and more pathogenic than those propagated in vivo when tested against the fifth larval instar of C fumiferana (Clem) and C occidentalis Freeman. The dose-responses (LD50 and LD95, expressed as occlusion bodies per larva) of C fumiferana larvae to in vitro-propagated OBs were 274 and 5785, respectively. The values of LD50 and LD95 to C occidentalis larvae were 19 and 118, respectively. There were no significant differences in pathogenicity or size when OBs propagated in C fumiferana larvae were tested against either insect species, nor were there significant differences for OBs propagated in C occidentalis larvae. The LD50 and LD95 of in vivo-produced OBs to C fumiferana were 925 and 61988, respectively. The LD50 and LD95 to C occidentalis were 50 and 453, respectively. OBs propagated in vitro had a mean volume of 13.13 microm3, whereas those propagated in vivo ranged from 0.84 to 1.41 microm3. The median survival time-responses (ST50) of fifth-instar C fumiferana or C occidentalis larvae to OBs propagated in vivo were not significantly different from those propagated in vitro at the dosage levels tested. Values of ST50 of C fumiferana larvae to in vitro- and in vivo-produced OBs at dosages causing less than 50% mortality rangedfrom 9.6 to 9.8 days post-inoculation (dpi), whereas a LD95 dose resulted in ST50 values ranging from 7.3 to 7.7 days. ST50 values of C occidentalis larvae at dosages causing less than 50% mortality ranged from 9.8 to 10.2 dpi, whereas a LD95 dose resulted in ST50 values ranging from 9.5 to 9.8 dpi. The median feeding cessation time-response (FT50) of fifth-instar C fumiferana larvae to OBs propagated in vitro (5.7 days) was not significantly different from the FT50 of those propagated in vivo in either insect species (5.3 and 5.7 days) at the dosage level tested (LD95). No significant differences in FT50 values were observed between OBs propagated in either larval host. The FT50 of C occidentalis larvae to OBs propagated in vitro (7.7 days) was not significantly different from that to those propagated in vivo in C occidentalis larvae (7.6days), but somewhat different (7.2 days) from that to those propagated in C fumiferana larvae. Results indicate that CfMNPV can be propagated in vivo in either C fumiferana or C occidentalis larvae (or sequentially through both) without alteration in infectivity, although the use of the CF-203 cell line yields the most biologically active OBs.     

Effect of environmental conditions on the seasonal and inter‐annual variability of small pelagic fish abundance off North‐West Africa: The case of both Senegalese sardinella

The objective of this study was to assess the effect of environmental variations on the abundance of Sardinella aurita and Sardinella maderensis in Senegalese waters in the upwelling system. Monthly data indicating the abundance of sardinella were first estimated from commercial statistics, using Generalized Linear Model from 1966 to 2011. Abundance indices (AIs) were then compared with environmental indices, at the local scale, a Coastal Upwelling Index (CUI) and a coastal Sea Surface Temperature (SST) index, and on a large scale, the North Atlantic Oscillation (NAO), the Atlantic Multidecadal Oscillation (AMO) and the Multivariate El Niño Southern Oscillation Index (MEI), using correlations and times series analyses. The results showed that the abundance of sardinella is determined by a strong seasonal pattern and inter‐annual fluctuations. The abundance of S. aurita peaked in spring and in autumn, whereas that of S. maderensis peaked in the warm season (July–September). The trend of the sardinella abundance was significantly correlated with the CUI, especially in autumn and spring. Interannual fluctuations of S. maderensis and S. aurita abundance are, respectively, driven by the precocity and the duration of the upwelling season that is attributed to distinct migration patterns. Both sardinella species also respond with a delay of around 4 years to the winter NAO index and the autumn CUI, and the AMO index, respectively, both related to migration patterns. The wide variations in sardinella biomass are caused by variations in environmental conditions, which should be considered in the implementation of an ecosystem‐based approach in sardinella stocks management.     

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.