The use of a real-time PCR primer/probe set to observe infectivity of Yersinia ruckeri in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and steelhead trout, Oncorhynchus mykiss (Walbaum).

Yersinia ruckeri is the causative agent of enteric redmouth disease (ERM), a common pathogen affecting aquaculture facilities and implicated in large losses of cultured fish. Fisheries scientists continue to gain a greater understanding of the disease and the pathogen by investigating methods of identification and pre- and post-infection treatment. In this study, a real-time PCR probe set for Y. ruckeri was developed to detect daily changes in the bacterial load during pathogen challenges. Two species of fish, Chinook salmon, Oncorhynchus tshawytscha, and steelhead trout, Oncorhynchus mykiss, were exposed to two strains of Y. ruckeri (Hag and SC) during bath challenges. A subset of fish was killed daily for 14 days, and the kidney tissue was biopsied to enumerate copies of pathogen DNA per gram of tissue. While Chinook exposed to either the Hag or SC strains exhibited similar pathogen loads, those exposed to the Hag strain displayed higher mortality (∼66%) than fish exposed to the SC strain (∼24% mortality). Steelhead exposed to the Hag strain exhibited a greater pathogen load and higher mortality (∼42%) than those exposed to the SC strain (<1% mortality). Steelhead challenged with either strain showed lower pathogen loads than Chinook. The study illustrates the efficacy of the probe set to enumerate Y. ruckeri bacterial growth in the kidneys of fish. Also, strains of Y. ruckeri display species-specific growth patterns that result in differential mortality and pathogen load.

Physiological condition differentially affects the behavior and survival of two populations of sockeye salmon during their freshwater spawning migration.

Recently, a segment of the Adams-Shuswap sockeye salmon (Oncorhynchus nerka) population initiated freshwater migration several weeks earlier than historically recorded, resulting in high mortality rates. The comigrating Chilko population maintained their historic river entry timing and did not experience elevated mortality. To test the hypothesis that population-specific differences in physiological condition would differentially influence behavior and survival when exposed to fisheries capture stress, we physiologically sampled individuals from both populations at the onset of the freshwater phase of their reproductive migration and tracked the remainder of their migrations using radio telemetry. Adams-Shuswap individuals had slower migration rates and were less likely to reach natal subwatersheds relative to Chilko individuals. Metabolic and osmoregulatory impairment was related to mortality for Adams-Shuswap individuals but not for Chilko individuals. Similarly, physiological condition correlated with migration rate for Adams-Shuswap but not Chilko fish. Survival to natal subwatersheds was 1.9 times higher for Chilko relative to Adams-Shuswap, a result that did not emerge until individuals approached natal subwatersheds several days after the stressor was applied. We conclude that physiological condition differentially affects the behavior and survival of these two populations, which may be a consequence of the early-entry phenomenon by a segment of the Adams-Shuswap population.

The influence of dissolved oxygen on winter habitat selection by largemouth bass: an integration of field biotelemetry studies and laboratory experiments.

In this study, field biotelemetry and laboratory physiology approaches were coupled to allow understanding of the behavioral and physiological responses of fish to winter hypoxia. The biotelemetry study compared dissolved oxygen levels measured throughout the winter period with continually tracked locations of nine adult largemouth bass obtained from a whole-lake submerged telemetry array. Fish habitat usage was compared with habitat availability to assess whether fish were selecting for specific dissolved oxygen concentrations. The laboratory study examined behavioral and physiological responses to progressive hypoxia in juvenile largemouth bass acclimated to winter temperatures. Results from the dissolved oxygen measurements made during the biotelemetry study showed high variance in under-ice dissolved oxygen levels. Avoidance of water with dissolved oxygen <2.0 mg/L by telemetered fish was demonstrated, but significant use of water with intermediate dissolved oxygen levels was also found. Results from the lab experiments showed marked changes in behavior (i.e., yawning and vertical movement) at <2.0 mg/L of dissolved oxygen but no change in tissue lactate, an indicator of anaerobic metabolism. Combined results of the biotelemetry and laboratory studies demonstrate that a dissolved oxygen content of 2.0 mg/L may be a critical threshold that induces behavioral responses by largemouth bass during the winter. In addition, the use by fish of areas with intermediate levels of dissolved oxygen suggests that there are multiple environmental factors influencing winter behavior.

Morphological correlates of swimming activity in wild largemouth bass (Micropterus salmoides) in their natural environment.

Individual variation in morphology has been linked to organismal performance in numerous taxa. Recently, the relationship between functional morphology and swimming performance in teleost fishes has been studied in laboratory experiments. In this study, we evaluate the relationship between morphology and swimming activity of wild largemouth bass (Micropterus salmoides) during the reproductive period, providing the first data derived on free-swimming fish not exposed to forced swim trials in the laboratory. Sixteen male largemouth bass were angled from their nests, telemetered, and subsequently monitored by a whole-lake acoustic hydrophone array with sub-meter accuracy. Additionally, eleven morphological measurements were taken from digital images of each fish. A principal components analysis of the morphological measurements described 79.8% of the variance. PC1 was characterized by measures of overall body stoutness, PC2 was characterized by measures of the length and depth of the caudal region, and PC3 characterized individuals with relatively large anterior portions of the body and relatively small caudal areas. Of these variables, only PC3 showed significant relationships to swimming activity throughout the parental care period. PC3 was negatively correlated with multiple measures of swimming activity across the parental care period. Furthermore, swimming performance of individual male bass was noted to be repeatable across the parental care period indicating that this phenomenon extends beyond the laboratory.

Effects of a dietary Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle.

Three experiments were conducted to examine the effects of an Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle. In Exp. 1, 120 crossbred steers were used in a randomized complete block design to evaluate the effects of roughage source (alfalfa hay vs. cottonseed hulls) and supplemental alpha-amylase at 950 dextrinizing units (DU)/kg of DM. Significant roughage source x alpha-amylase interactions (P < 0.05) were observed for performance. In steers fed cottonseed hulls, supplemental alpha-amylase increased ADG through d 28 and 112 and tended (P < 0.15) to increase ADG in all other periods. The increases in ADG were related to increased DMI and efficiency of gain during the initial 28-d period but were primarily related to increased DMI as the feeding period progressed. Supplemental alpha-amylase increased (P = 0.02) the LM area across both roughage sources. In Exp. 2, 96 crossbred heifers were used in a randomized complete block design with a 2 x 3 factorial arrangement of treatments to evaluate the effects of corn processing (dry cracked vs. high moisture) and supplemental alpha-amylase concentration (0, 580, or 1,160 DU/kg of DM). Alpha-amylase supplementation increased DMI (P = 0.05) and ADG (P = 0.03) during the initial 28 d on feed and carcass-adjusted ADG (P = 0.04) across corn processing methods. Longissimus muscle area was greatest (quadratic effect, P = 0.04), and yield grade was least (quadratic effect, P = 0.02) in heifers fed 580 DU of alpha-amylase/kg of DM across corn processing methods. In Exp. 3, 56 crossbred steers were used in a randomized complete block design to evaluate the effects of supplemental alpha-amylase (930 DU/kg of DM) on performance when DMI was restricted to yield a programmed ADG. Alpha-amylase supplementation did not affect performance when DMI was restricted. We conclude that dietary alpha-amylase supplementation of finishing beef diets may result in increased ADG through increased DMI under certain dietary conditions and that further research is warranted to explain its mode of action and interactions with dietary ingredients.

Long-term effects of low histidine intake on men.

The purpose of this 85-day study was to investigate the long-term effects of histidine depletion on nitrogen utilization in young adult men. A low nitrogen (6.3 g/day), low histidine (10 mg/day) amino acid diet was fed to seven men for 8 weeks. Mean nitrogen balance became negative at the end of the 8-week period. Free histidine in postabsorptive plasma and 24-hour urine decreased significantly during the first 2 weeks of the depletion and remained low and constant for the remaining 6 weeks. Hemoglobin concentration decreased somewhat, and serum iron concentration increased significantly during histidine depletion. Lean body mass, urinary N'-methylhistidine and total creatinine did not change significantly. On addition of histidine to the low histidine diet for 2 weeks, nitrogen retention became positive, plasma and urinary histidine returned to initial values, serum iron fell, and hemoglobin concentration slowly increased. These parameters remained unchanged in two control men fed the same diet supplemented with histidine (1.05 g/day) for 8 weeks. The results suggest that histidine is indispensable for young men consuming a low nitrogen diet.

Effects of dietary histidine and arginine on nitrogen retention of men.

Effects of dietary histidine and arginine on nitrogen retention were compared in six young men consuming for 1 week, each of six semipurified diets containing eight indispensable amino acids proportioned as in casein and 6.3 g nitrogen daily. Nonspecific nitrogen was either A) a mixture of six dispensable amino acids and arginine (diet 1) or arginine and histidine (diet 2) in casein proportions, or B) an isonitrogenous mixture of glycine and diammonium citrate alone (diet 3), with histidine (diet 4), arginine (diet 5), or histidine and arginine (diet 6). Nitrogen retention was significantly greater when the nonspecific nitrogen source was dispensable amino acids, arginine and histidine (diet 2) than when it was glycine and diammonium citrate (diet 3). mean balances were positive only when diets contained histidine (diets 2, 4, and 6). Histidine with arginine (diets 2 and 6) significantly improved nitrogen retention compared to arginine alone, but the balance, although positive, was not significantly improved when histidine was fed without arginine (diet 4). Urinary urea nitrogen confirmed these data. Indicators of erythrocyte status, plasma enzyme activities and proteins, and creatinine clearance were unaffected by diet. In summary, histidine supplementation of the low nitrogen diet improved total nitrogen utilization when arginine was present in the diet.

Effect of bile salts on hepatic phosphatidylcholine synthesis and transport into rat bile.

The effect of transport of micelle-forming and non-micelle-forming conjugated bile salts on phosphatidylcholine synthesis and transport into bile was studied in the ex vivo perfused rat liver. Single additions of sodium taurocholate, a good micelle-forming conjugated bile salt, caused an increase in bile flow associated with increased phosphatidylcholine and taurocholate concentration. The specific activity of phosphatidylcholine with respect to incorporated [1,2-(14)C]choline and [(3)H]methyl of l-[Me-(3)H]methionine was not significantly altered by the increased transport of phosphatidylcholine. The data suggested that bile phosphatidylcholine is synthesized to a great extent, although not exclusively, by phosphorylcholine glyceride transferase. Single additions of the glycine conjugate of dehydrocholate, a poor micelle-forming bile salt, caused an increase in bile flow comparable to that seen with sodium taurocholate administration. However, the concentrations of phosphatidylcholine in bile decreased. Thin-layer and gas-liquid chromatographic analyses of bile secreted before and after glycodehydrocholate administration revealed no significant increase in bile salt secretion other than the administered glycodehydrocholate. Investigations utilizing radiochemically pure [(14)C]glycine dehydrocholate revealed that increased bile flow after [(14)C]-glycine dehydrocholate administration occurs concomitantly with the secretion of 75-95% of the administered [(14)C]glycine dehydrocholate as a single peak into bile. Thus the increased bile flow without increased phosphatidylcholine concentration noted after glycodehydrocholate administration is due to transport of an intact, nonmetabolized, conjugated bile salt with poor micelle-forming properties. The data indicate that the formation of a bile salt-phosphatidylcholine micelle is responsible for phosphatidylcholine transport into bile.