Influence of Dietary Ascorbic Acid on the Immune Responses of Juvenile Korean Rockfish Sebastes schlegelii

Juvenile Korean Rockfish Sebastes schlegelii (length, 13.6 ± 1.4 cm [mean ± SD]; weight, 53.6 ± 4.2 g) were fed twice daily with diets containing varying levels of ascorbic acid (0, 50, 100, 200, and 400 mg/kg) for 4 weeks. Significant increases in daily weight and length occurred in fish fed more than 50 mg/kg ascorbic acid. The lysozyme activity of fish fed diets containing ascorbic acid was considerably increased in rockfish plasma at 400 mg/kg ascorbic acid and in kidney at over 50 mg/kg ascorbic acid. Total plasma immunoglobulin M (IgM) levels were markedly elevated in fish fed 400 mg/kg ascorbic acid. The results suggest that dietary ascorbic acid supplementation in juvenile rockfish can induce a significant increase in growth and in immunological features such as lysozyme activity and plasma IgM levels.

Received October 1, 2014; accepted May 2, 2015     

Effect of ellagic acid on some haematological,immunological and antioxidant parameters of rainbow trout (Oncorhynchus mykiss)

In this study, effect of ellagic acid on some haematological, immunological and antioxidant parameters in the blood and various tissues of rainbow trout (Oncorhynchus mykiss) were examined. Four groups of rainbow trout were fed experimental diets containing either no ellagic acid (control) or supplemented with ellagic acid at 50 mg/kg diet (EA‐50), 100 mg/kg diet (EA‐100) or 150 mg/kg diet (EA‐150) for 21 days. Samples of the blood and tissue (liver, kidney and spleen) were collected at the end of the experiment and analysed for their haematological profile (the red blood cell count, the haemoglobin concentration and the haematocrit level), immune response (the white blood cell count, the oxidative radical production (NBT activity), the total plasma protein and total immunoglobulin level) and oxidant/antioxidant status (the malondialdehyde level, the superoxide dismutase, catalase and glutathione peroxidase activity as well as the reduced glutathione concentration). The findings of this study demonstrated that ellagic acid had a positive effect on the haematological parameters, the immune response and the antioxidant enzyme activities of the fish.     

Developmental competence of eggs produced by rainbow trout Doubled Haploids (DHs) and generation of the clonal lines

Poor quality eggs produced by the fully homozygous doubled haploids (DHs) may impair generation of clonal lines in fish species. In the present research, gynogenetic development of rainbow trout (Oncorhynchus mykiss) was induced in eggs originated from the DH females. Eggs were activated with the UV‐irradiated grayling (Thymallus thymallus) spermatozoa and subjected to the high hydrostatic pressure (HHP) shock to provide diploid clonal individuals. Only two of four DH females produced eggs that were successfully activated by the irradiated spermatozoa and subsequently developed into the gynogenetic embryos. Survival rates of rainbow trout from the clonal lines equalled 21.5% and 19.8% during embryogenesis and decreased after hatching to 18.6% and 14.9%, respectively. Some of the dead rainbow trout clones collected between hatching and swim‐up stage were emaciated and exhibited spinal deformities including scoliosis. Provided results confirmed limited developmental competences of eggs produced by rainbow trout DH females. Clonal rainbow trout developing in such eggs exhibited reduced survival and increased frequency of the body abnormalities.     

The dose–response relation for the antinociceptive effect of morphine in a fish,rainbow trout

Jones, S. G., Kamunde, C., Lemke, K., Stevens, E. D. The dose–response relation for the antinociceptive effect of morphine in a fish, rainbow trout. J. vet. Pharmacol. Therap.  35 , 563–570. There have been suggestions that analgesics be used by fish researchers. But in the absence of dose–response data for morphine, this suggestion seems imprudent. The purpose of the present study was to develop a dose–response relationship in fish using six doses of morphine. The response (movement of the fins or tail) to a noxious stimulus (electrical shock to the face region) was monitored before and after a dose of morphine intraperitoneally (i.p.). The i.p. dose of morphine ED₅₀ in rainbow trout was 6.7 ± 0.8 mg/kg (n = 12 at each dose). The plasma morphine concentration EC₅₀ was 4.1 ± 1.5 mg/L. In a second experiment, rainbow trout tested with equal amounts of morphine and naloxone (30 mg/kg) showed that the antinociceptive effect of morphine was blocked by naloxone. It has been suggested that stress‐induced analgesia has been a confounding factor in some fish studies. However, plasma cortisol levels in our study indicated that stress was not a confounding factor in the present experiments. The ED₅₀ for morphine in fish was higher than that reported for humans or other mammals. Our observation showing a dose–response relation for morphine using a noxious stimulus supports arguments for its effectiveness as an antinociceptive drug in fish.     

The partial substitution of digestible protein with gelatinized starch as an energy source reduces susceptibility to lipid oxidation in rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax) muscle

We evaluated the influence of dietary gelatinized starch and protein on the fatty acid composition of muscle in rainbow trout and European sea bass and on the susceptibility of flesh to lipid peroxidation. The possibility that flesh peroxidation could be accounted for by lipogenesis and the deposition of fat was also explored. The inclusion of gelatinized starch in the diet of rainbow trout improved growth with respect to that observed in fish fed crude starch (P<.001). This was especially noticeable at the lowest concentration of dietary protein tested (P = .037); suggesting that gelatinized starch may partially replace protein in the production of energy without inducing a negative effect on growth. However, in European sea bass, the gelatinization of starch and dietary protein concentration showed no significant effect on final body weight. The intramuscular neutral lipid concentration of the sea bass was reduced by the gelatinization of dietary starch (P = .034). The highest dietary protein concentration increased the proportion of saturated fatty acids in the neutral (P = .0742) and polar (P = .0033) lipid fractions. The dietary inclusion of high levels of protein in rainbow trout led to a lower concentration of total (n-3) (P = .0457) and (n-6) (P = .0522) fatty acids and a higher concentration of total monounsaturated fatty acids (P = .0006). The inclusion of gelatinized starch led to a lower concentration of (n-3) fatty acids (P = .0034) and a higher concentration of saturated fatty acids (P = .0007). The polar fraction was hardly affected by the same treatment. A significantly lower susceptibility of the dorsal muscle to oxidation was observed in groups of European sea bass fed gelatinized starch (P<.01). A similar trend was observed in rainbow trout, although differences were not significant. The findings suggest that the digestible protein concentration of nutrient-dense diets for rainbow trout and European sea bass can be reduced with a beneficial effect on tissue lipid oxidation and no negative effects on growth and muscle composition.     

Effects of Supplemental Amino Acid-Chelated Trace Elements on the Immune Response of Rainbow Trout Subjected to Bacterial Challenge

Abstract

This study examined whether supplemental amino acid-chelated trace elements were effective in improving disease resistance in rainbow trout Oncorhynchus mykiss. Four semipurified diets containing trace elements from either sulfates or an amino acid chelate source were fed to the fish for 18 weeks. Subsequently, fish weighing about 20–40 g were challenged with β-hemolytic Streptococcus sp. The cumulative mortality was less in the chelate-supplemented groups but not significantly different from that of the sulfate groups. Antibody titer was significantly higher in the chelate-fed fish. Lysozyme activity and total immunoglobulin content were slightly more enhanced in the chelate-fed than in the sulfate-fed groups. Despite the increased antibody production, no distinct improvement in disease resistance against Streptococcus sp. was achieved by supplementing rainbow trout diets with amino acid-chelated trace elements. Further confirmatory studies focusing on the amounts of the trace elements used are needed to shed light on the influence of amino acid chelates on disease resistance.     

White Sturgeon as a Potential Vector of Infectious Hematopoietic Necrosis Virus

Abstract

Cell lines from white sturgeon Acipenser transmontanus were derived from peripheral blood cells, heart, and spleen. Incubated with infectious hematopoietic necrosis virus (IHNV) for 8 d at l5°C, these cell lines produced 0.7–53.2 plaque-forming units (PFU)/cell. Waterborne exposure of larval white sturgeons (60 d posthatch) to 10⁶ PFU/mL of IHNV resulted in 10% mortality 5–6 d postinfection, with virus concentrations consistently greater than 10⁵ PFU/g. A replicate group of larval white sturgeons that were sampled at different times post-IHNV exposure had no detectable virus at 24 h, but 72% of the fish had IHNV concentrations of 10²-10⁶ PFU/g when they were examined 2–9 d postinfection. Juvenile white sturgeons (mean weight, 35 g) immersed in or injected with IHNV exhibited no mortality, and virus was only detected immediately postexposure in just 25% of the fish tested. Juvenile white sturgeons fed either virus-free rainbow trout Oncorhynchus mykiss or dead IHNV-infected rainbow trout had no viable virus in their feces. Juvenile white sturgeons fed or exposed to IHNV failed to transmit the virus to cohabiting rainbow trout fry. These results suggest that IHNV can replicate in larval white sturgeons but presumably not in juveniles or adults. Virus neutralization activity was detected in serum from adult white sturgeons (4–6 years old) cultured with rainbow trout exposed to IHNV but not in white sturgeons kept in a pathogen-free environment and fed a manufactured diet. White sturgeon serum with IHNV-neutralizing activity was used to passively immunize rainbow trout, and it provided significant (P < 0.01) protection against IHNV challenge.     

Vitamin E and organic selenium enhances the antioxidative status and quality of chicken semen under high ambient temperature

1. The effects of supplemental dietary vitamin E and organic selenium (Se), and their combination, on improving semen quality characteristics and antioxidative status were investigated in cockerels exposed to high ambient temperature.

2. A total of 36 Egyptian local cross males, 40 weeks old, were housed individually in cages in an open-sided building (average daily temperature ranged from 33 to 36°C and relative humidity from 60 to 70%). Birds were divided randomly into 4 experimental treatments (n?=?9 each): (1) control (basal diet without any supplementation with vitamin E or Se); (2) vitamin E (basal diet +200?mg α-tocopherol acetate/kg diet); (3) Se (basal diet +0·3?mg organic Se/kg diet); and (4) vitamin E?+?Se (basal diet +200?mg α-tocopherol acetate/kg diet +0·3?mg organic Se/kg diet).

3. Under heat stress conditions, inclusion of vitamin E and/or organic Se in the diets enhanced the semen quality traits, including the spermatozoa count and motility, and reduced the percentage of dead spermatozoa.

4. A combination of 200?mg/kg vitamin E with 0·3?mg/kg organic Se reduced the thiobarbituric acid reactive substance concentration in seminal plasma samples to about 28% of the controls; and also enhanced the seminal plasma glutathione peroxidase activity by two-fold compared with controls.

5. It was concluded that dietary vitamin E in combination with organic Se has a synergistic effect in minimising lipid peroxidation and improving the antioxidative status in seminal plasma of the domestic fowl, which probably translated into enhanced spermatozoa count, motility and reduced percentage of dead spermatozoa under heat stress conditions.     

Influence of Seminal Plasma Antioxidants and Osteopontin on Fertility of the Arabian Horse

This study was designed to investigate enzymatic antioxidants’ activity and nonenzymatic antioxidants’ levels in seminal plasma of stallions and to relate them with season, age, and fertility of stallions. Fifty ejaculates were collected from six healthy Arabian stallions, 4-22 years old. Ejaculates were evaluated by conventional methods. Five milliliters of each semen sample was centrifuged, and the supernatant seminal plasma was stored at −20°C. Five antioxidants, in addition to osteopontin (OPN) and testosterone, were determined in stallion seminal plasma by using commercial enzyme-linked immunosorbent assay kits. Results revealed that uric acid, ascorbic acid, OPN, and testosterone concentrations and glutathione peroxidase (GPx) activity in stallions’ seminal plasma were high (P < .05) during spring. GPx activity was higher (P < .05) in age group B (11-18 years old) than in age group A (4-10 years old). The effect of stallions’ age on GPx activity in the fertility groups was highly significant (P < .01). OPN concentration was highest (P < .05) in age group A. Uric acid and OPN concentrations and GPx activity in stallions’ seminal plasma and percent of sperm motility were higher (P < .05) in fertility group III (>70%) than in fertility group I (<50%). However, ascorbic acid concentration, catalase activity and percentage of sperm abnormalities were lower (P < .05) in fertility group III than in fertility group I. It was concluded that season and stallion age may affect antioxidant defense systems in stallions’ seminal plasma. The impairment of seminal antioxidants and OPN could lead to low fertility.     

The Effects of Whirling Disease on Growth and Survival of Snake River Cutthroat and Colorado River Rainbow Trout Fingerlings

Abstract

Two sizes of fingerling Snake River cutthroat trout Oncorhynchus clarkii behnkei and Colorado River rainbow trout O. mykiss were raised at hatcheries testing negative for Myxobolus cerebralis and stocked into the Dolores and Cache la Poudre rivers from 1999 to 2001. Populations were resampled over a 2-year period to determine which species and size combination had the highest growth and survival rates. Fish were tested for M. cerebralis via polymerase chain reaction and pepsin?trypsin digest analyses. Growth and survival rates between the species and size groups were not significantly different in either river. In the Dolores River, annual survival for both species and sizes of fish combined ranged from 0.063 to 0.12. In the Cache la Poudre River, survival for both sizes of rainbow trout was 0.004; survival for cutthroat trout ranged from 0.182 to 0.53. Larger fish had higher growth rates than smaller fish, and cutthroat trout had higher rates than similar sizes of rainbow trout. In both rivers, a higher percentage of the rainbow trout sample was infected than in the cutthroat trout sample. Rainbow trout also had a higher mean number of spores per head than cutthroat trout, and small rainbow trout had higher spore counts than large rainbow trout. Survival rates for cutthroat trout in the Cache la Poudre River were the highest of any of the groups, suggesting a difference that is biologically significant. Raising fingerlings to sizes greater than 100 mm can improve poststocking survival. If rainbow trout are stocked into contaminated waters, raising fingerlings to a larger size does not appear to improve growth or survival rates. Stocking rainbow trout in the spring could maximize growth rates but will expose fish to greater triactinomyxon densities, resulting in higher intensities of infection.     

Metabolism and residue depletion of albendazole and its metabolites in rainbow trout, tilapia and Atlantic salmon after oral administration

Metabolic and residue depletion profiles of albendazole (ABZ) and its major metabolites in three fish species, rainbow trout, tilapia and Atlantic salmon are reported. Based on these profiles, similarities (or dissimilarities) between species will determine the potential to group fish species. ABZ at 10 mg/kg body weight was incorporated into fish food formulated in a gelatin base or in gel capsule and fed as a single dose to six fish from each species. Rainbow trout were held three each in a partitioned 600-L tank. Tilapia and Atlantic salmon were housed in separate 20-L tanks. Samples of muscle with adhering skin were collected at 8, 12, 18, 24, 48, 72, and 96 h postdose from trout kept at 12 degrees C, at 4, 8, 12, 24, 48, 72, 96, 120, and 144 h postdose from tilapia kept at 25 degrees C and at 8, 14, 24, 48, 72, and 96 h postdose from Atlantic salmon kept at 15 degrees C. The samples were homogenized in dry ice and subjected to extraction and cleanup procedures. The final extracts were analyzed for parent drug ABZ and its major metabolites, albendazole sulfoxide (ABZ-SO), albendazole sulfone (ABZ-SO2) and albendazole aminosulfone using high-performance liquid chromatography with fluorescence detection. ABZ was depleted by 24 h in trout and tilapia and by 48 h in salmon; ABZ-SO, a pharmacologically active metabolite, was depleted by 48 h in tilapia, by 72 h in rainbow trout and was present until 96 h in salmon; and low levels of ABZ-SO2 and albendazole aminosulfone, both inactive metabolites, were detectable at least till 96 h in all three fish species.     

Lipid and protein oxidation levels in spermatozoa and seminal plasma of Asian Elephants (Elephas maximus) and their relationship with semen parameters

Peroxidation damage to spermatozoa and seminal plasma has an important role in sperm quality. Thus, the objective of this study was to determine the levels of lipid and protein oxidation in spermatozoa and seminal plasma of Asian elephants (Elephas maximus) with varying percentage of progressive motility. Lipid and protein oxidation was measured by the thiobarbituric acid‐reactive species (TBARS) assay and the 2, 4‐dinitrophenylhydrazine (DNPH) carbonyl groups assay, respectively. Fresh semen samples were collected from Asian elephants and classified according to the percentage of motile spermatozoa into good (>60%) and poor (≤20%) motility. Results revealed that seminal plasma malondialdehyde (MDA) and seminal plasma protein carbonyls (PCs) were significantly higher in poor motility than in good motility (p < .05). The MDA and PC levels in seminal plasma were negatively correlated with the percentages of progressive motility (p < .05). In addition, the negative correlation between sperm concentration and seminal plasma MDA level was investigated (p < .05). The sperm viability was also negatively correlated with sperm PC level (p < .05). This study indicated that lipid and protein oxidation has deleterious effect on semen quality of Asian elephants.     

Effects of Ergosan on the expression of cytokine genes in the liver of juvenile rainbow trout (Oncorhynchus mykiss) exposed to enteric red mouth vaccine

The effects of administration of the immunomodulator Ergosan, an algal extract containing alginic acid, in juvenile rainbow trout (Oncorhynchus mykiss) exposed to AquaVac vaccination, were tested. Juveniles treated with Ergosan, 95 days after the beginning of first solid feeding and control fish fed solely on commercial diet, were vaccinated by immersion in AquaVac solution. The time-course of the effects of vaccination on liver immunorelated gene modulation and on the tolerance to stress manipulation connected with the vaccination was investigated. Liver and plasma sampling was performed at the following times: T=pre-vaccination, T0=5min, T1=2h, T2=8h, T3=24h, T4=48h and T5=72h post-vaccination. Interleukin-1beta (IL-1beta), interleukin-8 (IL-8), tumor necrosis factor alpha 2 (TNF alpha 2) and heat shock protein 70 (Hsp70) gene expression in trout liver was monitored by real-time PCR using Acidic Ribosomal Phosphoprotein P0 (ARP) as internal standard. The evaluation of the plasma cortisol levels was performed by EIA. In AquaVac-vaccinated fish, both the gene expression of Hsp70 and the plasma cortisol levels during the time-course were significantly (P<0.05) lower in Ergosan-treated fish with respect to control, indicating the positive role of Ergosan on handling stress tolerance. This study also demonstrated the stimulatory properties of Ergosan on cytokine genes expression involved in innate immune response: liver IL-1beta, IL-8 and TNF alpha 2 gene expression was significantly (P<0.05) higher in trout fed on Ergosan compared to control, indicating a positive role of this feed additive in improving the immune responsiveness to AquaVac vaccine.     

Distribution of Myxobolus cerebralis in Salmonid Fishes in Montana

Abstract

Over an approximately 2-year period, 20,974 fish (trout and other salmonid species) from 230 separate waters (creeks, rivers, lakes, reservoirs, ponds, hatcheries, and irrigation ditches) within 21 of the 22 major drainages in Montana were examined for Myxobolus cerebralis. Nine of the major river drainages have waters containing infected fish: Beaverhead, Big Hole, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the south fork of the Flathead River, Jefferson, Madison, Missouri above the Marias River, and Yellowstone above the Bighorn River. The Beaverhead, Clark Fork above the Bitterroot River, Jefferson, Madison, and Missouri above the Marias River have the greatest number of waters containing fish infected with M. cerebralis. Comparisons of infection levels (number of pooled samples that contain fish infected with M. cerebralis) between species among these drainages show significantly lower levels of infection in brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss in the Missouri above the Marias River drainage and significantly higher levels of infection of rainbow trout in the Jefferson. Comparisons of differences in infection levels between drainages among species show that, in the Beaverhead, Clark Fork above the Bitterroot River, and Madison, infection levels in brown trout are significantly higher than in rainbow trout. This is partially attributed to losses of juvenile rainbow trout because of M. cerebralis infection, leading to biased samples. Histopathologic studies showed lesions were consistently less severe in brown trout than other species and occurred in a different location (gill arches versus ventral calvarium). In six of the nine affected drainages (Beaverhead, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the South Fork, Jefferson, and Madison), infected fish were found at or near the time that intensive sampling was initiated in the spring of 1995. In the three remaining affected drainages (Missouri above the Marias River, Yellowstone above the Bighorn River, and the Big Hole), infected fish were not identified until at least 15 months after the initiation of widespread testing. This indicates that in the first six drainages listed above, the infection was well established prior to 1995 but spread into the last three drainages in the ensuing months. Methods of transmission and the sources of infection are unknown, although the absence of infected fish in state, private, and federal hatcheries in Montana indicates hatchery fish from these sources are not likely to be responsible.     

Susceptibility of Selected Inland Salmonids to Experimentally Induced Infections with Myxobolus cerebralis,the Causative Agent of Whirling Disease

Abstract

Laboratory exposures to the infectious stages (triactinomyxons) of Myxobolus cerebralis demonstrated a range of susceptibility to whirling disease among four species of inland salmonids. Replicate groups of each species were exposed to two concentrations of triactinomyxons, a low dose (100–200 per fish) and a high dose (1,000–2,000 per fish). Exposed fish were evaluated for clinical signs, for severity of microscopic lesions at 35 d, 2 and 5 months, and for spore concentrations in the head cartilage at 5 months. A standard strain of rainbow trout Oncorhynchus mykiss matched for age served as a susceptible species control. Rainbow trout, westslope cutthroat trout O. clarki lewisi, Yellowstone cutthroat trout O. clarki bouvieri, and bull trout Salvelinus confluentus were susceptible to M. cerebralis infections. Clinical signs, including radical swimming (“whirling”) and black tails, were observed at 7 weeks postexposure among rainbow and cutthroat trout challenged at 3 weeks of age. Clinical signs were rare among bull trout exposed at an age of 4 weeks and absent among rainbow and cutthroat trout exposed at 3 months posthatch. Most rainbow, cutthroat, and bull trout were found to be infected when examined at 5 months postexposure. The most severe microscopic lesions among infected fish at 5 months postexposure were found among rainbow trout. Cutthroat trout had less severe lesions, bull trout had mild infections, and no evidence of infection was found among Arctic grayling Thymallus arcticus. Mean spore concentrations among infected fish correlated with the severity of microscopic lesion scores. Rainbow trout had mean concentrations of spores in head cartilage reaching 10⁶, whereas more resistant species such as bull trout had 10⁴ spores; no spores were found among Arctic grayling at 5 months postexposure.     

Effect of dietary Ulva and Spirulina on weight loss and body composition of rainbow trout, Oncorhynchus mykiss (Walbaum), during a starvation period

A study was conducted to determine the effect of dietary algal supplementation on weight loss and proximate composition in rainbow trout, Oncorhynchus mykiss, during starvation. An algae-free control diet (C) and four experimental diets, including varying levels of raw Ulva meal (5% = U5; 10% = U10) and Spirulina meal (5% = S5; 10% = S10) were formulated. Fish were fed to satiation for 12 weeks and then subjected to a 3-week starvation period. Body weight, viscerosomatic index (VSI, %), hepatosomatic index (HSI, %), dress-out (DO, %) and chemical composition of carcass were analysed at the end of each starvation week. The accumulated weight loss of the fish fed dietary algae was almost 50% less than the control group (p < 0.05) after 2 and 3 weeks starvation. No significant differences in weight loss were observed among the fish fed different algal diets during the starvation period. Total body protein and ash contents remained constant in all groups throughout the starvation period. Fish in all treatments lost a significant level of total lipids and changes of liver fatty acid profiles were also observed. Results indicate that low level inclusion of algae in aquafeeds may have economical advantages in terms of reducing weight loss in fish when subjected to a short-term fasting period, that sometimes is necessary after a heavy rainfall or when fish stocks are held back to meet market demand.     

Differences between Plasma and Serum Samples for the Evaluation of Blood Chemistry Values in Rainbow Trout,Channel Catfish,Hybrid Tilapias,and Hybrid Striped Bass

Abstract

Blood chemistry analytes are determined for fish from either serum or plasma samples. Serum and plasma are similar in that they both represent the fluid component of blood; however, plasma contains clotting factors that are not present in serum. This study was conducted to determine whether the type of sample—plasma or serum—had an effect on measured blood analytes in rainbow trout Oncorhynchus mykiss, channel catfish Ictalurus punctatus, hybrid tilapia Oreochromis spp., and hybrid striped bass (striped bass Morone saxatilis × white bass M. chrysops). Paired plasma and serum samples were analyzed for the following standard biochemical analytes: total protein, albumin, globulin, creatinine, total bilirubin, alkaline phosphatase, aspartate aminotransferase, sodium, potassium, chloride, calcium magnesium, phosphorus, glucose, and cholesterol. For all four taxa, values for potassium were lower in the serum and magnesium and phosphorus values were higher in the serum. Glucose values were lower in the serum from rainbow trout, hybrid striped bass, and channel catfish; whereas cholesterol values were higher in the serum of rainbow trout, channel catfish, and hybrid tilapias. The differences observed between serum and plasma were distinct from changes occurring with hemolysis and, therefore, do not represent release of erythrocyte constituents. The differences most likely represent metabolic utilization of blood constituents while the blood was clotting. This work indicates that plasma should be used preferentially to serum for biochemical analysis because analyte levels determined from serum may not accurately reflect those found in circulating blood.     

Effects of Acute Stress on Blood Clotting and Yeast Killing by Phagocytes of Rainbow Trout

Abstract

The effects of different levels of acute stress on blood parameters in rainbow trout Oncorhynchus mykiss were examined. Experimental fish were exposed to a combination of stressors, including handling, confinement, and thermal shock, and were bled 30 s to 30 min later. Control fish were sampled quickly without intentional stress. Two activities of blood were investigated: (1) whole-blood clotting (coagulation) times and (2) the ability of heparinized plasma to influence the survival of yeast soaked in this plasma and subsequently exposed to trout phagocytes. The speed of blood clotting increased with increasing levels of stress, whereas blood from many resting fish failed to clot. Yeast incubated in plasma from stressed fish before exposure to pronephric phagocytes experienced a greater reduction in colony-forming units (cfus) than yeast incubated in plasma from resting fish. Different levels of stress did not induce significant differences in reduction of yeast cfus. These data support the idea that acute stress may actually stimulate primary defense mechanisms rather than suppress them. Furthermore, whole-blood clotting times seemed to be reliable indicators of the severity of stressors to which fish were exposed. We postulate that factors appearing in blood very quickly, such as catecholamines and specific plasma proteins, play important roles in both the killing activities of phagocytes and the blood-clotting process.     

Pathological effects of Arcobacter cryaerophilus infection in rainbow trout (Oncorhynchus mykiss Walbaum)

Arcobacter cryaerophilus was isolated from naturally infected rainbow trout (Oncorhynchus mykiss Walbaum), and its pathogenicity was tested by intramuscular injection into 40 healthy 1-year-old rainbow trout at 16 degrees C. The lethal dosage of 50% end point (LD50) for A. cryaerophilus was calculated 2.25 x 10(4) viable cells. Experimental infection caused deaths with gross clinical abnormalities such as degenerated opercula and gills, liver damage, haemorrhagic kidney and serous fluid in swollen intestines. The counts of A. cryaerophilus in kidney, liver and gills of experimentally infected fish ranged from 1.59 x 10(10) colony forming units (cfu)/g to 7.41 x 10(12) cfu/g. The means of erythrocyte (RBC) count, haematocrit level, serum cholesterol, triglyceride, albumin and total protein concentrations in the blood of the experimentally infected rainbow trout group were significantly lower than in the healthy fish. Leukocyte (WBC) counts of the experimentally infected rainbow trout were significantly higher than those of healthy fish. The present work shows that the selected blood characteristics may be good indicators of response to infections in rainbow trout.     

欧亚鲈幼鱼饲料中维生素C适宜含量的研究

选用初始体质量为(7.46±0.29)g的欧亚鲈幼鱼为试验对象,投喂6种不同水平的维生素C(0、50、100、200、400和800 mg/kg)的试验饲料,饲养8周,探讨欧亚鲈幼鱼饲料维生素C的适宜含量。结果表明,饲料中维生素C添加量从0.0升高到100 mg/kg时,鱼体增重率呈上升趋势,添加量为100 mg/kg显著高于对照组(P<0.05),但添加量继续增大,对鱼体增重率无显著变化(P>0.05);经折线模型分析表明,欧亚鲈鱼种达到最大生长时,饲料中的维生素C的最低添加量为93.58 mg/kg。欧亚鲈肝脏中维生素C含量随着饲料中维生素C(0～200 mg/kg)的增加而显著升高(P<0.05),但添加量继续升高,则肝脏中维生素C达到饱和状态,经折线分析表明,肝脏中维生素C达到饱和时,饲料中的维生素C的最低添加量为149.27 mg/kg。急性胁迫前,各组血糖之间无显著差异(P>0.05),急性胁迫后,血糖升高峰值最低和恢复最快的是饲料中维生素C为400～800 mg/kg。在本试验条件下,欧亚鲈幼鱼达到最大生长、肝脏维生素C饱和及抗拥挤胁迫的维生素C适宜添加量分别为93.58、149.27 mg/kg和400 mg/kg。