Isolation and preliminary characteristics of β-N-acetylglucosaminidase in the sperm of Siberian sturgeon (Acipenser baerii) and rainbow trout (Oncorhynchus mykiss)

The aim of this study was to characterize the enzyme β- N -acetyglucosaminidase (β-NAGase) in the milt and spermatozoa extracts from Siberian sturgeon and rainbow trout. After ion exchange chromatography one protein peak showed β-NAGase activity in sturgeon milt plasma and sperm extracts of both species. Surprisingly, two protein peaks showing β-NAGase activity were found in rainbow trout milt plasma. The molecular mass of β-NAGase was estimated by gel filtration as 127 kDa for rainbow trout spermatozoa, 271 kDa for sturgeon spermatozoa, and 74 kDa for milt plasma from both species. The kinetic parameters were determined for milt plasma and sperm extracts. The optimum pH of the β-NAGases was 3.8 for sturgeon milt plasma, 4.4 for sturgeon sperm extract, and 4.4–4.8 for milt plasma and sperm extract from rainbow trout. K _m value of the β-NAGases was 0.212, 0.563, 0.779 m m for sturgeon milt plasma, sturgeon sperm extract or rainbow trout extract, respectively. The β-NAGase from sperm extracts in both species showed 100% activity even after incubation at 56°C by 20 min, whereas its activity was decreased to 23% in sturgeon milt plasma and to 2% in trout milt plasma.     

Temperature comparisons for antibody production in vitro by plaque-forming cells from trout, Salmo gairdneri (Richardson), and mice

Anterior kidney and splenic cells were taken from rainbow trout and splenic cells from BALB/c mice immunized with a T-dependent (sheep red blood cells) or T-independent (DNP-Ficoll) antigen. The cells were incubated at different temperatures in Jerne plaque assays (direct or passive haemolytic plaque assays). The optimum numbers of in vitro plaque-forming cells (PFC) after incubation with homologous complement were directly correlated with normal body temperatures of the respective species. The optimum incubation temperature was 37°C for mouse cells and 10°C for fish cells. Incubation of mouse cells at lower temperatures of 30, 20, 10, 4 or 0°C appeared to yield a direct line reduction in numbers of PFC. Trout cells developed significantly fewer PFC at 4 and 20°C and none at 30°C or above; however, significant numbers still appeared at 0°C. More PFC per million white blood cells were obtained from the anterior kidney; however, related to temperatures, no differences in development of numbers of PFC could be seen between the spleen and anterior kidney cells of trout. When the incubation time was lengthened for both trout and mouse cells held at low temperatures, the numbers of PFC approached those of the cells incubated at the optimum temperatures for 10 h.     

THE INFLUENCE OF TEMPERATURE ON THE SURVIVAL OF SEVERAL SPECIES OF FISH IN LOW TENSIONS OF DISSOLVED OXYGEN

The resistance to low tensions of dissolved oxygen over periods ranging from less than a day to a week was determined for rainbow trout ( Salmo gairdnerii Richardson), perch ( Perca fluviatilis L.), roach ( Rutilus rutilus (L.)), and mirror carp, a domestic form of the common carp ( Cyprinus carpio L.) at 10°, 16° and 20°C., tench ( Tinea tinea (L.)) at 10° and 16°C., dace ( Leuciscus leuciscus (L.)) at 16° and 20°C., chub ( Squalius cephalus (L.)) at 20°C., and bleak ( Alburnus alburnus (L.)) at 16°C.
At each temperature period of survival decreased with fall in oxygen tension.
Rise in temperature between 10° and 20°C. reduced resistance to lack of oxygen of all species except rainbow trout, in which there was no significant difference between resistance at 16° and 20°C. Rise in temperature between 10° and 16°C. did, however, lower resistance considerably.
Tench were the most resistant to lack of oxygen and, in general, trout were the most susceptible. At 20°C. mirror carp required a higher tension of oxygen to enable them to survive for a week than did rainbow trout, but for shorter periods could withstand much lower tensions than rainbow trout; at 10° and 16°C. their resistance was intermediate between that of tench and rainbow trout.     

The effects of heat shock and acclimation temperature on hsp70 and hsp30 mRNA expression in rainbow trout: in vivo and in vitro comparisons

Heat shock (25° C) of 10° C-acclimated rainbow trout Oncorhynchus mykiss led to increases in heat shock protein 70 (hsp70) mRNA in blood, brain, heart, liver, red and white muscle, with levels in blood being amongst the highest. Hsp30 mRNA also increased with heat shock in all tissues with the exception of blood. When rainbow trout blood was heat shocked in vitro , both hsp70 and hsp30 mRNA increased significantly. In addition, these in vitro experiments demonstrated that blood from fish acclimated to 17° C water had a lower hsp70 mRNA heat shock induction temperature than did 5° C acclimated fish (20 v. 25° C). The hsp30 mRNA induction temperature (25° C), however, was unaffected by thermal acclimation. While increases in hsp70 mRNA levels in blood may serve as an early indicator of temperature stress in fish, tissue type, thermal history and the particular family of hsp must be considered when evaluating stress by these molecular means.     

Characteristics of the spawning migrations of brown trout, Salmo trutta L., and rainbow trout, S. gairdneri Richardson, in Great Lake, Tasmania

Upstream spawning migrations of mature brown trout, S. trutta , and rainbow trout, S. gairdneri , were studied in Liawenee Canal, Great Lake from 1949 to 1985. Brown trout migrations normally occurred from early April to mid-May and rainbow trout from late August to early November. In 1983, 16 425 brown trout and 1338 rainbow trout passed through a fixed upstream diversion trap. Brown trout spawning migrations occurred predominantly over the temperature range 6–10° C, while rainbow trout migrated predominantly over the range 5–11° C. Migrations peaked at water temperatures of 7.6°C (males) and 7.8°C (females) for brown trout, and 8.3°C (males) and 9.6°C (females) for rainbow trout. Rainbow trout migrations occurred at high flow conditions and were positively correlated with canal flow increases, while brown trout migrated under low canal flow. Mean length, weight and condition of rainbow trout of both sexes decreased significantly during migrations. Female brown trout decreased in weight and condition but not in length; male brown trout did not change in condition despite decreases in both length and weight during migrations. Overall sex ratio was 2:1 (female:male) for both species, with the relative proportion of male fish decreasing as migrations progressed. Age composition changed during migrations; dominant age classes were 3 < 4 < 5 + years for both species. Comparison of length, weight, condition and age revealed minor changes during the 37-year period 1949–1985.     

Body temperature and seawater adaptation in farmed Atlantic salmon and rainbow trout during prolonged chilling

The core temperature of the rainbow trout Oncorhynchus mykiss (3·5 kg) dropped to 1·0° C during the first 6 h of chilling at 0·5° C, remained stable until 24 h, and dropped significantly to 0·7° C after 39 h. Blood plasma osmolality increased and muscle moisture content decreased gradually with increasing chilling time. After 39 h of chilling, the rainbow trout experienced 40 mosmol l^-1 higher blood plasma osmolality and 2·8% less muscle moisture content compared with initial values. In the Atlantic salmon Salmo salar (5·3 kg), core temperature dropped to 1·3° C and blood plasma osmolality increased significantly during the first 6 h of chilling at 0·5° C, but remained relatively stable throughout the rest of the experimental period. After 39 h of chilling, the salmon experienced 20 mosmol l^-1 higher blood plasma osmolality and 0·5% less muscle moisture content compared with initial values. In rainbow trout muscle moisture content was inversely related to blood plasma osmolality indicating reduced seawater adaptation with increasing hours of chilling. No such relationship was observed in the Atlantic salmon. Hence, changes in plasma osmolality and muscle moisture in the Atlantic salmon do not indicate osmoregulatory failure since the new levels, once established, were maintained throughout the chilling time.     

Bile Pigments in the Bile of Freshwater Fish,Rainbow Trout,Tilapia, and Pejerrey

Bile pigments in the bile of the rainbow trout, Salmo gairdneri, tilapia, Tilapia nilotica, and pejerrey, Odonthetes bonariensis, were analyzed by HPLC and TLC.

Major bile pigments of rainbow trout and pejerrey were bilirubin glucuronides and ditauro-bilirubin, respectively. Ditaurobilirubin was not detected in rainbow trout and bilirubin glucuronides were scarcely found in pejerrey. Biliverdin seemed to be the sole bile pigment in tilapia, but it was a minor component in the other fish. These results are in accord with the previous reports in which the diversity of bile pigment composition was demonstrated in some fish.     

Toxicity of crude extracellular products of Aeromonas hydrophila in tilapia, Tilapianilotica

Extracellular products (ECP) secreted from Aeromonas hydrophila with haemolytic andproteolytic activity were studied with respect to temperature and time of incubation as well as thelethal toxicity on tilapia, Tilapia nilotica . The highest production of the haemolysin productwas achieved when Aer. hydrophila was grown at 35°C for 30 h. Tilapia erythrocytewas found to be more susceptible than sheep erythrocyte for determining the haemolytic activity.The haemolytic activity against tilapia erythrocyte was completely inactivated after heating theECP at 60°C for 10 min or 55°C for 15 min. The proteolytic activity was maximized whenthe bacterium was grown at 30°C for 36 h. Complete inactivation of the protease enzyme wasperformed after heating the ECP at 80°C for 10 min or 70°C for 15 min. Aeromonashydrophila was found to produce haemolytic and proteolytic exotoxin lethal to tilapia (LD₅₀ 2·1 × 10⁴ cell/fish), as well as heat stable unknown virulent factors thatwere responsible for 20% mortality. The lethality of ECP was decreased by heating andcompletely inactivated by boiling at 100°C for 10 min.     

Temperature-dependence of cardiac output and regional blood flow in rainbow trout, Salmo gairdneri Richardson

Cardiac output, blood flow distribution and regional perfusion were determined in free-swimming rainbow trout acclimated to 6, 12 and 18°C, using the indicator dilution and microsphere methods. Cardiac output (ml min⁻¹ kg⁻¹) increased linearly with increasing temperature, while circulation time decreased. Blood flow distribution (% of cardiac output) to the spleen, liver, kidney, gall bladder and gastro-intestinal tract was significantly reduced at 18°C relative to 6°C-acclimated fish. White muscle received the largest fraction of cardiac output, and blood flow distribution to white muscle increased significantly with increasing acclimation temperature. Blood perfusion (ml h⁻¹ g⁻¹) of various organs and red muscle was not influenced by acclimation temperature, while white muscle perfusion increased with increasing temperature. These results demonstrate physiological adaptation of the cardiovascular system of rainbow trout to changes in acclimation temperature.     

三种鲟科鱼类和两种鲑科鱼类卵黄免疫球蛋白性质的比较

从施氏鲟(Acipenser schrencki)、小体鲟(Acipenser ruthenus)、西伯利亚鲟(Acipenser baeri)、哲罗鲑(Hucho taimen)和金鳟(Oncorhynchus mykiss)五种鱼类卵黄中分离、纯化Ig,并对其分子结构进行了初步研究。结果表明:五种鱼类卵中Ig的分子量分别为施氏鲟524kD、小体鲟468kD、西伯利亚鲟475kD、哲罗鲑为490kD,金鳟498kD,其Ig重链的相对分子量相同,约为97kD。轻链的相对分子量各不相同,施氏鲟为27kD,小体鲟28.5kD,西伯利亚鲟30kD,哲罗鲑28.5kD和金鳟16kD。其中西伯利亚鲟、施氏鲟和小体鲟Ig等电点约为5.85,哲罗鲑和金鳟的Ig等电点约为6.55,其蛋白质结构上具有一定的相似性,且均为糖蛋白     

Comparative stable isotope analyses of green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) in the San Francisco estuary

Green sturgeon (Acipenser medirostris) and white sturgeon (A. transmontanus) are closely related, sympatric species that inhabit the San Francisco estuary. Green sturgeon have a more marine life history but both species spawn in the Sacramento River and reside for some duration in San Francisco Bay. These sturgeons are of conservation concern, yet little is known about their dietary competition when they overlap in space and time. To examine evidence of dietary differentiation, we collected whole blood and blood plasma from 26 green sturgeon and 35 white sturgeon in San Francisco Bay. Using carbon and nitrogen stable isotope analyses, we compared their relative trophic levels and foraging locations along the freshwater to marine gradient. Sampling blood plasma and whole blood allowed comparison of dietary integration over shorter and longer time scales, respectively. Plasma and whole blood δ¹³C values confirmed green sturgeon had more marine dietary sources than white sturgeon. Plasma δ¹⁵N values revealed white sturgeon fed at lower trophic levels than green sturgeon recently, however, whole blood δ¹⁵N values demonstrated the two species fed at the same trophic level over longer time scales. Larger individuals of both species had higher δ¹³C values than smaller individuals, reflecting more marine food sources in adulthood. Length did not affect δ¹⁵N values of either species. Isotope analyses supported the more marine life history of green than white sturgeon and potentially highlight a temporary trophic differentiation of diet between species during and preceding the overlapping life stage in San Francisco Bay.     

Inhibition of β-N-acetylglucosaminidase by acetamide affects sperm motility and fertilization success of rainbow trout (Oncorhynchus mykiss) and Siberian sturgeon (Acipenser baerii)

β-N-Acetylglucosaminidase (β-NAGase) is an enzyme found in the sperm acrosome of numerous animal species including fish. Fish spermatozoa differ in their morphology including acrosome or acrosomeless aquasperm in chondrostean (e.g., sturgeon) and teleostean (e.g., rainbow trout). It has been shown that β-NAGase exists with high activity in both eggs and sperm of these species. The present study shows the potency of β-NAGase in fertilization. In rainbow trout, increase in sperm motility parameters (VAP and MOT) were observed in the presence of acetamide, an inhibitor for β-NAGase. In contrast, sperm motility parameters (VCL, VSL, VAP, MOT, and PRG) were reduced on the Siberian sturgeon in the presence of acetamide. The inhibition of the activity of β-NAGase in rainbow trout spermatozoa was led to a reduction in the number of fertilized eggs from 79% to 40%, whereas in sturgeon no change was observed in fertilization. Moreover, inhibition of β-NAGase in both spermatozoa and eggs of trout and sturgeon resulted in significant decrease in fertilization rate from 79% to 1% in rainbow trout and from 84% to 12% in Siberian sturgeon. Our research proves that β-NAGase can play a significant role in the fertilization process in teleosteans.     

Effects of ambient colour on colour preference and growth of juvenile rainbow trout Oncorhynchus mykiss (Walbaum)

Colour preference of individual juvenile rainbow trout Oncorhynchus mykiss was tested at 1 and 12° C, and also at 12° C after a 42 day growth experiment under white, blue, green, yellow or red ambient colour. All experiments were carried out under controlled laboratory conditions and the preference was assessed by the location of the fish in a preference tank with four chambers. Rainbow trout showed a preference for blue and green at 1° C and for green at 12° C. After the growth experiment the fish reared in blue tanks preferred blue and green but green was the most preferred colour for the fish reared in green, yellow and red tanks. Yellow and especially red chambers were avoided, irrespective of the ambient colour during the growth trial. The final mass of fish reared in the red aquaria was significantly smaller than that of the fish in green tanks. In addition, when the data of the preference tests were correlated with the data of the growth experiment using mean values of the four tested colours, a very good linear relationship was observed between the preference ( i.e. visit frequency in coloured compartments) and growth rate as well as food intake. When considering the results both from the preference and growth trials it is suggested that green is the best environmental colour for rearing juvenile rainbow trout while rearing in a red environment cannot be recommended.     

Oxygen uptake by rainbow trout blood, Salmo gairdneri

Oxygen uptake rate by rainbow trout red cells was measured in the temperature range 5° to 40° C. Between 5° and 20° C there was a steady increase in rate and at 20° C a maximum rate of 2.5 μl O₂/min/ml red cells occurred, followed by a decline in the rate as the temperature increased. A morphological investigation of the red cell was conducted and possible functions of the nucleus in fish red blood cells are discussed.     

The level and composition of free fatty acids in the plasma of freshwater fish in a post-absorptive condition

The level and fatty acid composition of plasma free fatty acids (FFA) were determined in four important aquaculture teleost species: goldfish (Carassius auratus), common carp (Cyprinus carpio), rainbow trout (Oncorhynchus mykiss) and tilapia (Oreochromis mossambicus). Plasma was obtained from animals after a 3 day fast in order to study the mobilization of fatty acids in a post-absorptive condition. Plasma [FFA] amounted to 0.95 in goldfish, 0.77 in both carp and rainbow trout and 0.30 μmol/ml in tilapia. The fatty acid composition was dominated by mono-unsaturates in goldfish and carp, whereas saturates were the main fatty acids in tilapia. The largest amount of PUFA was observed in rainbow trout (41%) which was considerably higher than in the other species (27–29%). The interspecies differences of plasma FFA levels and patterns are discussed in relation to the site of lipid storage, mobility and other factors.     

Influence of temperature on silver accumulation and depuration in rainbow trout

To assess the influence of water temperature on silver uptake, rainbow trout Oncorhynchus mykiss ( c . 50 g; held at 13° C) were exposed to 0·1 μM AgNO₃ in ion‐poor water for 1 week at 4 and 16° C without previous temperature acclimation. To assess the influence of temperature on elimination of previously accumulated Ag, rainbow trout were exposed to 0·1 μM AgNO₃ in ion‐poor water for 1 week at 12° C, then were randomly divided amongst two Ag‐free water containers, differing only in temperature (3 and 16° C), for 2 months. In the uptake study greater accumulation of Ag was seen in the gills, plasma and especially the livers and bile of 'warm' rainbow trout (16° C) compared to 'cold' rainbow trout (4° C), which can be explained by the higher metabolic rates of the warmer fish. In the depuration study there was no net elimination of Ag from the livers and bile but there was biphasic elimination of Ag from the gills and plasma of 'warm' and 'cold' fish, but with few differences between them. This indicated that temperature‐dependent processes were less important in Ag elimination than in Ag uptake. Toxicokinetic modelling of Ag uptake by livers indicated four‐fold greater uptake of Ag by 'warm' rainbow trout compared to 'cold' rainbow trout (one compartment uptake model). Elimination of previously accumulated Ag from the plasma was best fitted by a two compartment rate‐constant based model, with approximately half the plasma Ag load eliminated within 24 h, followed by slower elimination of Ag over 2 months.     

THE EFFECT OF TENSION OF DISSOLVED OXYGEN ON THE TOXICITY OF UN-IONIZED AMMONIA TO SEVERAL SPECIES OF FISH

The resistance to rapidly lethal concentrations of un-ionized ammonia ranging from about 2.0 to 8.8 p. p. m. N was determined for rainbow trout ( Salmo gairdnerii Richardson), perch ( Perca fluviatilis L.), roach ( Rutilus rutilus (L.)), and gudgeon ( Gobio gobio (L.)) in tensions of dissolved oxygen 53.4 and 96.7% of air-saturation value at 15.2°C.
The relation between concentration of un-ionized ammonia and period of survival from a few hours to 13 days was investigated for rainbow trout in two tensions of dissolved oxygen, 45.7 and 100.3% of air-saturation value, at 20.1°C.
Period of survival of all the species tested decreased with rise in concentration of un-ionized ammonia.
Decrease in tension of dissolved oxygen increased the toxicity of un-ionized ammonia to all species except gudgeon, for which there was no significant difference in period of survival at the two levels of oxygen.
The effect of oxygen tension on period of survival was greatest in the lowest concentrations of un-ionized ammonia.
Resistance of perch and roach to lack of oxygen was not significantly affected, but that of rainbow trout was reduced significantly, by the presence of a small concentration of ammonia (not toxic to trout within at least 13 days).     

Temperature effects on anaesthesia with clove oil in six temperate‐zone fishes

The potential use of clove oil (eugenol) as an anaesthetic for Atlantic salmon Salmo salar , brown trout Salmo trutta , rainbow trout Oncorhynchus mykiss , whitefish Coregonus lavaretus , perch Perca fluviatilis and roach Rutilus rutilus was examined at 5, 10, 15 and 20° C using three anaesthetic concentrations (varying from 20 to 200 mg l⁻¹) at each temperature. Substantial species differences in sensitivity to clove oil were observed, even amongst congeners, and there may be some disadvantages (slow recovery and possibly mortality) with using clove oil for 0+ year whitefish and at low temperatures for perch and roach.     

Comparison of the activity of carp and rat β-actin gene regulatory sequences in tilapia and rainbow trout embryos

A comparative study on the level of expression of lacZ reporter constructs driven by equivalent carp and rat β-actin regulatory sequences was carried out in embryos of tilapia and rainbow trout. DNA was microinjected into fertilised tilapia and rainbow trout eggs and the embryos/fry were assayed at various developmental stages for β-galactosidase expression. We provide evidence to demonstrate that the carp β-actin promoter/lacZ reporter gene is expressed at higher levels than the equivalent rat β-actin construct in both species. © 1996 Wiley-Liss, Inc.     

Effect of starvation and refeeding on digestive enzyme activities in sturgeon (Acipenser naccarii) and trout (Oncorhynchus mykiss)

The digestive enzyme activities were determined in Adriatic sturgeon and rainbow trout during starvation and refeeding period. Overall, the digestive enzyme activities are affected in the same sense in both species. The protease and lipase activities were decreased later than amylase activity. Even after 1 month of starvation, both species would be prepared to digest protein and lipids in an effective way. After 72 days of starvation, the digestive machinery of the sturgeon and of the trout shows an altered capacity to digest macronutrients. The capacity to digest proteins and lipids, after 60 days of refeeding, begins to become re-established in sturgeon and trout. In contrast, in this period, the capacity to digest carbohydrates remains depressed in both species.