Piscine reovirus in wild and farmed salmonids in British Columbia,Canada: 1974–2013

Piscine reovirus (PRV) was common among wild and farmed salmonids in British Columbia, western Canada, from 1987 to 2013. Salmonid tissues tested for PRV by real‐time rRT‐PCR included sections from archived paraffin blocks from 1974 to 2008 (n = 363) and fresh‐frozen hearts from 2013 (n = 916). The earliest PRV‐positive sample was from a wild‐source steelhead trout, Oncorhynchus mykiss (Walbaum), from 1977. By histopathology (n = 404), no fish had lesions diagnostic for heart and skeletal muscle inflammation (HSMI). In some groups, lymphohistiocytic endocarditis affected a greater proportion of fish with PRV than fish without PRV, but the range of Ct values among affected fish was within the range of Ct values among unaffected fish. Also, fish with the lowest PRV Ct values (18.4–21.7) lacked endocarditis or any other consistent lesion. From 1987 to 1994, the proportion of PRV positives was not significantly different between farmed Atlantic salmon, Salmo salar L. (44% of 48), and wild‐source salmonids (31% of 45). In 2013, the proportion of PRV positives was not significantly different between wild coho salmon, Oncorhynchus kisutch (Walbaum), sampled from British Columbia (5.0% of 60) or the reference region, Alaska, USA (10% of 58).     

Effects of day and night temperature on photosynthesis,antioxidant enzyme activities,and endogenous hormones in tomato leaves during the flowering stage

To study the effects of day and night temperature difference (DIF) on tomato growth, a controlled experiment using Solanum lycopersicum L., cv. Jinguan 5 was conducted. The daily mean temperature (T_m) was maintained at 18°C and 25°C, and the DIF was set at 0°C, 6°C, and 12°C. The results indicated that chlorophyll a (Chl a) and chlorophyll b (Chl b) gradually increased as DIF rose. At 18°C T_m, the carotenoid content reached a maximum at 12°C DIF. The Chl a/Chl b, net photosynthetic rate (P_N), photosynthetic rate at irradiation saturation (P_max), stomatal conductance (g_s), intercellular CO₂ concentration (C_i), stomatal limitation value, the maximum assimilation rate (A_max), apparent quantum efficiency (A_q), carboxylation efficiency (C_e), superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), gibberellin A₃ (GA₃), and indole-3-acetic acid (IAA) were highest, while malondialdehyde (MDA) was lowest at 6°C DIF. At 25°C T_m, P_N, P_max, A_max, A_q, C_e, g_s, C_i, CAT, POD, GA₃, IAA, and zeatin reached the maximum under 6°C DIF, while SOD and MDA reached the maximum under 12°C DIF. Furthermore, the morphological index peaked at 6°C DIF under 18°C and 25°C T_m. The results suggested that 6°C DIF improved the growth and development of tomato during the flowering stage. ABBREVIATIONS: Aq – apparent quantum efficiency; Amax – the maximum assimilation rate; CAT – Catalase; Chl a(b) – chlorophyll a(b); Ca – ambient CO₂ concentration; Ce – carboxylation efficiency; Ci – intercellular CO₂ concentration; DIF – difference between day temperature (TD) and night temperature(TN); FM – fresh mass; gs – stomatal conductance; GA₃ – gibberellin A₃; IAA – indole-3-acetic acid; Ls – stomatal limitation value; MDA – malondialdehyde; Pmax – photosynthetic rate at irradiation saturation; PN – net photosynthetic rate; POD – peroxidase; ROS – reactive oxygen species; SOD – superoxidedismutase; Tm – daily mean temperature; ZT – zeatin.     

An assay for quantitative virulence in Rhynchosporium commune reveals an association between effector genotype and virulence

Detailed knowledge of the evolutionary genetics of virulence is needed to understand and predict host–pathogen dynamics. This study used a virulence assay based on digital image analysis and treated virulence as a quantitative rather than a binary trait. Such quantitative data may better reflect the genetic underpinning of virulence in many pathogen systems and provide better resolution in statistical investigations. A greenhouse experiment based on a common garden design was conducted to measure virulence (% of leaf area covered by lesions) of 126 genetically distinct isolates of the barley scald pathogen, Rhynchosporium commune, originating from nine field populations around the world. Virulence in this pathosystem was found to be a quantitative trait with a continuous distribution in all populations. By comparing population genetic differentiation for virulence and neutral microsatellite markers (i.e. a Q_ST/G_ST comparison), evidence that virulence is under stabilizing selection across populations was found. Heritability values were high and ranged from 0·52 to 0·96 with a mean heritability of 0·84. Virulence was positively correlated with spore production as predicted by the trade‐off theory of virulence evolution. Furthermore, an association analysis between virulence and sequence haplotypes of three known necrosis‐inducing effector genes (NIP1, NIP2 and NIP3) revealed a significant effect of NIP2 haplotypes and NIP1 deletions. Overall, the results support a quantitative model for virulence in the R. commune–barley pathosystem and very high evolutionary potential for this trait.     

Evidence of diadromy in the French Polynesian Kuhlia malo (Teleostei: Percoidei) inferred from otolith microchemistry analysis

Abstract – Strontium/Calcium (Sr/Ca) and Barium/Calcium (Ba/Ca) ratios were measured in 60 otoliths of the French Polynesian flagtail Kuhlia malo. Both elemental ratios were needed to correctly distinguish residence in marine, brackish and fresh water. High Sr/Ca and low Ba/Ca around the nucleus of all otoliths provided evidence of marine residency during the early life of all individuals. At about 0.5 mm from the core, Ba/Ca increased rapidly and remained high in all otoliths. High and unstable Sr/Ca ratios were attributed to estuarine residency, whereas low and stable Sr/Ca ratios were attributed to freshwater residency. These chemical signatures inferred that following recruitment to rivers, some fish remained in the estuary, while others moved upstream. Some individuals shared their time between fresh and brackish waters later in life. The findings of this study have important implications for the protection of nearshore, brackish and freshwater environments for the conservation of this French Polynesian diadromous species.     

Comparison of dietary essential amino acid requirements determined from group‐housed versus individually‐housed juvenile bluegill,Lepomis macrochirus

Two 60‐day experiments were conducted sequentially to determine (i) lysine requirement of juvenile bluegill, Lepomis macrochirus based on the dose–response method, (ii) requirements for other essential amino acids (EAAs) using whole‐body amino acid profile and (iii) whether differences in growth rates of group‐housed versus individually‐housed bluegills lead to different lysine requirement levels because of the presence and absence, respectively, of social hierarchies. Seven, semi‐purified, experimental diets (isonitrogenous, isocaloric) were prepared to contain graded levels of digestible lysine (10–31 g kg⁻¹). Experiment‐1 involved group‐housed bluegills (approximately 27 g, n = 10 fish/chamber, 4 chambers/diet) whereas experiment‐2 involved individually‐housed bluegills (approximately 30 g, n = 1 fish/chamber, 14 chambers/diet). Fish were fed twice daily to apparent satiation. Bluegill growth responses in both experiments generally improved (P < 0.05, anova ) with increasing dietary lysine levels from 10 to 16 g kg⁻¹, and then levelled off with further increase in lysine level (P > 0.05). Optimal dietary lysine level (digestible basis) was estimated to be 15 g kg⁻¹ based on broken‐line regression analyses of relative growth rate and feed conversion ratio with no differences being observed between the two rearing methods. Determined dietary requirement levels for other EAAs ranged from 2.4 g kg⁻¹ (tryptophan) to 15.3 g kg⁻¹ (leucine).     

High dietary energy level stimulates growth hormone receptor and feed utilization in large Atlantic salmon (Salmo salar L.) under hypoxic conditions

This study examines how appetite and growth regulation of Atlantic salmon are affected by low dissolved oxygen (LO) and dietary digestible energy levels (DE: high [HE] vs. low [LE]). Long‐term exposure to LO resulted in a reduced feed intake, growth, digestible protein and fat retention efficiencies and increased feed conversation ratio and plasma ghrelin concentrations (p < .05) compared to high dissolved oxygen (HO). Pair‐feeding of rations based on the feed intake of the LO groups, but fed at HO, resulted in a 50% growth improvement in HE diet groups. This suggests that the poor growth under LO was not entirely caused by the reduced feed intake. Salmon adapted to LO by increased haemoglobin concentrations, while osmoregulation was affected by increased plasma chloride concentrations (p < .05). Plasma ghrelin concentration was unaffected by DE (p > .05). Growth regulation was affected by the HE diet, with increased liver and muscle growth hormone receptor ghr1 mRNA (p < .05), regardless of oxygen level. The growth depression due to low oxygen appears to be related to higher metabolic costs, while higher DE upregulates the GH‐IGF system at the ghr1 level and found to be beneficial for growth, feed intake, oxyregulation and osmoregulation under hypoxia.     

Identification of Neofabraea species causing bull's eye rot of apple in Poland and their direct detection in apple fruit using multiplex PCR

Based on partial sequence analysis of the β‐tubulin gene, 19 isolates of fungi causing bull's eye rot on apple in Poland were classified into species: Neofabraea alba, N. perennans and N. kienholzii. To the authors’ knowledge, the detection of N. kienholzii is the second in Europe and the first in Poland. Species affiliation of these fungi was confirmed by a new species‐specific multiplex PCR assay developed on the basis of previously published methods. The new protocol allowed for the specific identification of bull's eye rot‐causing species, both from pure cultures and directly from the skin of diseased or apparently healthy apples. In 550 samples of diseased fruits collected from nine cold storage rooms located in three regions of Poland, in 2011 and 2012, N. alba was detected as the predominant species causing bull's eye rot, occurring on average in 94% of the tested samples. Neofabraea perennans was found in a minority of apple samples, N. kienholzii was found only in two apple samples, while N. malicorticis was not detected in any sample tested. In tests on 120 apparently healthy fruits, only N. perennans was detected in a single sample. The results of genetic diversity analyses of bull's eye rot‐causing fungi based on the β‐tubulin gene sequence and an ISSR (inter‐simple sequence repeat) PCR assay with two primers were consistent, showing the expected segregation of tested isolates with respect to their species boundaries. However, the genetic distance between N. perennans and N. malicorticis was very low, as reported previously.