Upper and Lower Temperature Tolerance Limits for Juvenile Red Drums from Texas and South Carolina

Abstract

Upper and lower temperature tolerance limits, reported as temperatures lethal to 50% of the test fish (LT50s), were determined for juveniles of red drum Sciaenops ocellatus obtained from Texas and South Carolina brood stock. This information on red drum thermal tolerance limits was used to determine whether a more northerly distributed population exhibits greater cold tolerance and could therefore be used to improve success of stocking programs. A 96-h temperature tolerance test was performed on juveniles (71–155 mm total length) acclimated to water temperatures ranging from 12.0 to 28.0°C at a salinity of 20‰. Test fish were transferred directly into a series of high-temperature (27.0–37.0°C) or low-temperature (2.0–16.0°C) water baths. The upper LT50 for Texas red drums acclimated to 28.0°C was within 1.1 °C of that calculated for South Carolina red drums acclimated to the same temperature (upper LT50s were 35.7 and 34.6°C). Upper LT50s for fish acclimated to 20.0 and 12.0°C were also similar between the two stocks. Likewise, lower LT50s were similar between the two stocks of red drums: as determined from fish acclimated to 14.0 and 28.0°C, ranges of lower LT50s were 2.8–9.0°C for the Texas stock and 2.69.3°C for the South Carolina stock. The lower LT50 could not be determined for red drums acclimated to 12.0°C, because only three fish died. No consistent differences in temperature tolerance limits were found between the two hatchery stocks, precluding any recommendation to stock one group instead of the other.     

Temperature and mass scaling affect cutaneous and pulmonary respiratory performance in a diving frog

Global climate change is altering patterns of temperature variation, with unpredictable consequences for species and ecosystems. The Metabolic Theory of Ecology (MTE) provides a powerful framework for predicting climate change impacts on ectotherm metabolic performance. MTE postulates that physiological and ecological processes are limited by organism metabolic rates, which scale predictably with body mass and temperature. The purpose of this study was to determine if different metabolic proxies generate different empirical estimates of key MTE model parameters for the aquatic frog Xenopus laevis when allowed to exhibit normal diving behavior. We used a novel methodological approach in combining a flow-through respirometry setup with the open-source Arduino platform to measure mass and temperature effects on 4 different proxies for whole-body metabolism (total O₂ consumption, cutaneous O₂ consumption, pulmonary O₂ consumption, and ventilation frequency), following thermal acclimation to one of 3 temperatures (8°C, 17°C, or 26°C). Different metabolic proxies generated different mass-scaling exponents (b) and activation energy (E_A) estimates, highlighting the importance of metabolic proxy selection when parameterizing MTE-derived models. Animals acclimated to 17°C had higher O₂ consumption across all temperatures, but thermal acclimation did not influence estimates of key MTE parameters E_A and b. Cutaneous respiration generated lower MTE parameters than pulmonary respiration, consistent with temperature and mass constraints on dissolved oxygen availability, SA:V ratios, and diffusion distances across skin. Our results show that the choice of metabolic proxy can have a big impact on empirical estimates for key MTE model parameters.     

Metabolic rate and body temperature of an African sunbird,Nectarinia Chalybea: daily rhythm and the effect of ambient temperature

The oxygen consumption (VO₂) of the lesser double-collared sunbird, Nectarinia chalybea. was measured at ambient temperatures (Ta) from 7 to 35°C. The diel variation in body temperature (Tb) and wet thermal conductance (C) was also determined. The sunbirds (mean mass 8,36 g ± S.E. 0.21 g) showed a pronounced diel cycle of VO₂, with a mean reduction of 50% at night. Basal metabolic rate was estimated to be 3.23 ml O₂/g.h, which is 88% of the predicted value for a passerine bird of this size. There was a mean reduction in Tb of 3.6°C at night. Both day and night values of Tb decreased with decreasing Ta, as did individual values of C. Winter breeding in the south-west Cape is energetically stressful for lesser double-collared sunbirds, but they have the physiological capacity to reduce their energy expenditure substantially at night.     

Optimal Dosage of Erythromycin Thiocyanate in a New Feed Additive to Control Bacterial Kidney Disease

Abstract

An experimental feed additive containing erythromycin thiocyanate was formulated into fish feeds and tested to determine the optimal dosage and length of administration to treat acute infections of Renibacterium salmoninarum in chinook salmon Oncorhynchus tshawytscha. Trials were conducted on groups of yearling salmon acclimated and held in the laboratory at 10°C or 14°C in both the winter and spring. Parametric and nonparametric statistical analyses were used to assess survival rates during the tests and to evaluate the condition of fish alive at the completion of the trials. Survival was highest in trials of fish held at 10°C and among those fish that consumed higher quantities of erythromycin for 28 d of continuous therapy. Because portions of the daily rations were more likely to be refused when target dosages exceeded 100 mg/kg body weight, particularly in winter conditions, we recommend a standard therapeutic dosage of 100 mg/kg for 28 d.     

Temperature determines the shift of thermal neutral zone and influences thermogenic capacity in striped hamsters

The thermoneutral zone (TNZ) reflects the adaptation of mammals to their natural habitat. However, it remains unclear how TNZ shifts in response to variations in ambient temperature. To test the hypothesis that ambient temperature plays a key role in determining TNZ variations between seasons, we measured metabolic rate, body temperature, and cytochrome c oxidase (COX) activity of several visceral organs in striped hamsters (Cricetulus barabensis) either acclimated to semi-natural conditions over a year, or subjected to a gradual decrease in mean temperature from 30 ± 1°C to −15 ± 1°C. The TNZ range in striped hamsters differed seasonally, with a wider TNZ and a lower lower-critical temperature in winter compared to summer. The hamsters showed a considerable leftward shift of lower-critical temperature from 30°C to 20°C after the ambient temperature of acclimation from 30°C down to −15°C, whereas the upper-critical temperature of TNZ remained fixed at 32.5°C. The resting metabolic rate in thermoneutral zone (RMRt), nonshivering thermogenesis (NST), and COX activity of brown adipose tissue, liver, skeletal muscle, brain, and kidneys, increased significantly in hamsters acclimated at lower ambient temperatures. Following acute exposure to 5°C and −15°C, hamsters acclimated to 32.5°C had significantly lower maximal NST and lower serum thyroid tri-iodothyronine (T₃) levels compared to those kept at 23°C. These findings suggest that acclimation to the upper-critical temperature of TNZ impairs the hamsters’ thermogenic capacity to cope with extreme cold temperature. Reduced ambient temperature was mainly responsible for the leftward shift of TNZ in striped hamsters, which reflects the adaptation to cold environments.     

Effects of bacterial strains on the development of the ripening flavor of cured pork loins

To determine the effect of bacteria on the development of the ripening flavor of cured meat, pork loins were cured with pickles containing 8 or 16% sodium chloride at 0 and 8°C. The bacterial flora, total free amino acid content and total free fatty acid content in the cured loins, sensory properties of cooked pork loins and the relationships between the proteolytic and lipolytic activities of isolates and curing conditions were investigated. Desirable bacteria, including salt‐tolerant bacteria and lactic acid bacteria, were predominant under the condition of 8% sodium chloride in the pickles and a curing temperature of 8°C. The total free amino acid content and total free fatty acid content at a curing temperature of 8°C were higher than those at 0°C. Cooked pork loins cured in pickles containing 8% sodium chloride at a temperature of 8°C for more than 7 days were preferred in terms of color, flavor and taste. Before the curing procedure, gram‐negative bacteria (Vibrio, Acinetobacter, Pseudomonas and Enterobacteriaceae) were predominant in the pork loins. During the curing period, the numbers of viable gram‐positive bacteria (Micrococcus, Staphylococcus and Pediococcus) increased and the numbers of viable gram‐negative bacteria decreased. Strains of Micrococcus and Staphylococcus with proteolytic and/or lipolytic activities in cured meat also increased during the curing period and were more predominant in the pork loins cured in pickles containing 8% sodium chloride and at a curing temperature of 8°C than in pork loins cured in pickles containing 16% sodium chloride and at a curing temperature of 0°C. The actions of these bacteria were thought to be important factors affecting the flavor of the cured pork. Micrococcus and Staphylococcus with proteolytic activity might contribute to the development of the ripening flavor of ham. The results of the present study together with the results of further investigations on the relationships of the enzyme activities of Micrococcus and Staphylococcus in cured meat with a preferable flavor would be useful for establishing a novel effective method for using bacteria to produce ham of a high quality.     

Responses of Blue Catfish and Channel Catfish to Environmental Nitrite

Abstract

In nitrite-exposure experiments, percent methemoglobin, plasma nitrite concentration, and plasma chloride ion concentration were compared between channel catfish Ictalurus punctatus and blue catfish I. furcatus exposed to sublethal levels of nitrite for 48 h at 25°C. In nitrite-recovery experiments, fish exposed to elevated environmental nitrite for 12 h were transferred to freshwater, and blood characteristics were monitored during the 24-h recovery period. Blue catfish appeared to be more resistant to environmental nitrite than channel catfish. Methemoglobin levels (percent of hemoglobin in methemoglobin form) were significantly lower in blue catfish than in channel catfish. Maximum plasma nitrite concentrations were 137 mg NO₂-/L plasma in blue catfish and 164 mg NO₂-/L plasma in channel catfish. Percent methemoglobin and plasma nitrite concentration were closely correlated. Plasma chloride decreased initially with exposure to nitrite but quickly returned to control levels. Blue catfish exposed to nitrite at 10°C required 1 week to recover when placed in nitrite-free water. The methemoglobin reductase enzyme apparently functioned at a slow rate in fish acclimated to cold temperatures.     

Longevity of Bolbophorus damnificus Infections in Channel Catfish

Abstract

The digenean Bolbophorus damnificus infects commercial channel catfish Ictalurus punctatus, causing mortality, lower feed consumption, and reduced growth in surviving fish. The purpose of this study was to determine the length of time for which B. damnificus prodiplostomulum metacercariae (juvenile trematode stage that infects fish) would remain viable (parasite appearing to be intact or exhibiting movement) in channel catfish. Fish (n = 210) were infected with molecularly confirmed B. damnificus cercariae harvested from naturally infected marsh rams-horn snails Planorbella trivolvis. During the first sampling (at 20 d postinfection), 8.3 ± 3.6 metacercariae/fish (mean ± SD) were found in the host muscle and visceral organs. The channel catfish were then acclimated to a water temperature of either 18°C or 28°C. After 11 months, 6.8 ± 3.5 and 5.9 ± 3.0 metacercariae/fish were found in groups held at 18°C and 28°C, respectively. The mean number of parasites per fish did not significantly differ between fish held at the two temperatures and did not significantly decline over time at either temperature. Fish examined from 13 to 30 months postinfection all contained viable metacercariae that were morphologically and molecularly identified as B. damnificus. At 18 months, 12 metacercariae (of which 11 were intact and 10 displayed movement) were found in the one fish sampled; at 30 months, the last fish sampled contained three intact metacercariae (one displayed slight movement). Our results indicate that B. damnificus metacercariae can remain viable in channel catfish for at least an 18–30-month production cycle during which they have the potential to affect fish growth; in addition, infected fish may serve as intermediate hosts for these metacercariae for at least 2.5 years postinfection.

Received July 14, 2010; accepted March 6, 2011     

Pharmacokinetics of orbifloxacin in crucian carp (Carassius auratus) after intravenous and intramuscular administration

The pharmacokinetics of orbifloxacin was studied after a single dose (7.5 mg/kg) of intravenous or intramuscular administration to crucian carp (Carassius auratus ) reared in freshwater at 25°C. Plasma samples were collected from six fish per sampling point. Orbifloxacin concentrations were determined by high‐performance liquid chromatography with a 0.02 μg/ml limit of detection, then were subjected to noncompartmental analysis. After intravenous injection, initial concentration of 5.83 μg/ml, apparent elimination rate constant (λ_z) of 0.039 hr^?1, apparent elimination half‐life (T_1/2λz) of 17.90 hr, systemic total body clearance (Cl) of 75.47 ml hr^?1 kg^?1, volume of distribution (Vz) of 1,948.76 ml/kg, and volume of distribution at steady‐state (Vss) of 1,863.97 ml/kg were determined, respectively. While after intramuscular administration, the λ_z, T _1/2λz, mean absorption time (MAT ), absorption half‐life (T _1/2ka), and bioavailability were determined as 0.027 hr^?1, 25.69, 10.26, 7.11 hr, and 96.46%, respectively, while the peak concentration was observed as 3.11 ± 0.06 μg/ml at 2.0 hr. It was shown that orbifloxacin was completely but relatively slowly absorbed, extensively distributed, and slowly eliminated in crucian carp, and an orbifloxacin dosage of 10 mg/kg administered intravenously or intramuscularly would be expected to successfully treat crucian carp infected by strains with MIC values ≤0.5 μg/ml.     

Heat production and dissipation in a South African diurnal murid Lemniscomys griselda

Heat production measured as oxygen consumption (V_O2 and heat dissipation calculated as the means of the overall thermal conductance were studied in a South African diurnal murid Lemniscomys griselda. This species maintains its body temperature (T_b) well at around 36,5 ± 1,0 °C over ambient temperatures (T_a) ranging between 5 and 34 °C. The thermoneutral zone lies between T _a = 30-34 °C. When exposed to temperatures below the low critical point L. griselda at rest conforms to Newton's law of cooling. Non-shivering thermo- genesis (NST) was studied as a response to an injection of noradrenaline (sc). Following the administration of 1,5 mg noradrenaline per kg body mass to an anaesthetized animal, an increase of 329% in VO₂ was recorded. The distribution pattern of L. griselda in comparison with that of Rhabdomys pumilio, the more common diurnal murid species in South Africa, is discussed.     

Effects of Water Temperature on Experimental Transmission of Dermal Sarcoma in Fingerling Walleyes

Abstract

The effects of temperature on experimental transmission of dermal sarcoma, a spontaneous tumor of adult walleyes Stizostedion vitreum to fingerling walleyes was determined. Test temperatures were 10, 15, and 20°C. Three-month-old walleyes were inoculated intramuscularly with a cell-free filtrate from a dermal sarcoma collected from an adult walleye in the spring of the year. Tumors were first grossly visible in fish held at 15°C at 8 weeks postinoculation. At 12 weeks postinoculation, all fish were euthanized and examined for presence of tumors. Tumor transmission was most successful at 15°C, followed by that at 20°C; many fish maintained at these temperatures had grossly visible tumors. Although the majority offish held at 10°C also had tumors, tumors in these fish developed to a lesser degree than those observed in fish held at the higher temperatures.     

The effect of inhalant anesthetic and body temperature on peri-anesthetic serum concentrations of transdermally administered fentanyl in dogs

Objectives To determine whether moderate hypothermia during anesthesia significantly affects the serum concentration of transdermally delivered fentanyl and whether halothane or isoflurane affect these concentrations. Study Design Randomized cross‐over experimental trial. Animals Six mature, healthy Beagles (three males, three females) weighing 10.6 ± 0.43 kg. Methods A 50‐µg hour^?1 fentanyl patch was applied 36 hours prior to anesthesia. Anesthesia was induced at time 0 (t = 0). Each dog received four treatments: isoflurane + normothermia (ISO‐NORM), isoflurane + hypothermia (ISO‐HYPO), halothane + normothermia (HAL‐NORM), and halothane + hypothermia (HAL‐HYPO). Dogs were intubated and maintained at 1.5 times MAC. Animals in the hypothermia treatments were cooled to 35 °C during anesthesia. Serum fentanyl analysis was performed at ?36, ?24, ?12, 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 18, and 26 hours. Direct arterial blood pressures and arterial blood gases were monitored. Results The mean body temperatures (±SEM) during the anesthetic period for the four treatments were: ISO‐NORM = 37.7 ± 0.07 °C, ISO‐HYPO = 35.8 ± 0.1 °C, HAL‐NORM = 37.7 ± 0.06 °C, and HAL‐HYPO = 35.8 ± 0.13 °C. The mean (±SEM) serum fentanyl concentrations (SFC) for both hypothermia treatments were significantly lower than baseline concentrations at t = 1 hour and persisted for the duration of anesthesia for the ISO‐HYPO treatment but only from t = 1 to 2 hours for the HAL‐HYPO treatment. Serum fentanyl concentrations returned to baseline within one hour of the end of anesthesia, regardless of body temperature. There were no significant differences between treatments for systolic or diastolic blood pressure but mean blood pressures were higher during normothermia versus hypothermia during the last hour of anesthesia. Conclusions and clinical relevance Hypothermia during inhalation anesthesia produced a significant reduction in SFC using transdermal administration and was more protracted with isoflurane than halothane anesthesia. While significant reductions in SFC occurred, the SFC were still within the range believed to confer analgesia.     

Determination of sulfadiazine,trimethoprim, and N4‐acetyl‐sulfadiazine in fish muscle plus skin by Liquid Chromatography–Mass Spectrometry. Withdrawal‐time calculation after in‐feed administration in gilthead sea bream (Sparus aurata L.) fed two different diets

This study presents a depletion study for sulfadiazine and trimethoprim in muscle plus skin of gilthead sea bream (Sparus aurata L.). N⁴‐acetyl‐sulfadiazine, the main metabolite of sulfadiazine (SDZ), was also examined. The fish were held in seawater at a temperature of 24–26 °C. SDZ and trimethoprim (TMP) were administered orally with medicated feed for five consecutive days at daily doses of 25 mg SDZ and 5 mg TMP per kg of fish body weight per day. Two different diets, fish oil‐ and plant oil‐based diets, were investigated. Ten fish were sampled at each of the days 1, 3, 5, 6, 8, 9, 10, and 12 after the start of veterinary medicine administration. However for the calculation of the withdrawal periods, sampling day 1 was set as 24 h after the last dose of the treatment. Fish samples were analyzed for SDZ, TMP, and acetyl‐sulfadiazine (AcSDZ) residues by liquid chromatography–mass spectrometry. SDZ and TMP concentrations declined rapidly from muscle plus skin. Considering a maximum residue limit of 100 μg/kg for the total of sulfonamides and 50 μg/kg for TMP residues in fish muscle plus skin, the withdrawal periods of the premix trimethoprim‐sulfadiazine 50% were calculated as 5 and 6 days, at 24–26 °C, in fish oil (FO) and plant oil (PO) groups, respectively. The investigation of this work is important to protect consumers by controlling the undesirable residues in fish.     

Yolk‐sac larval development of the substrate‐brooding cichlid Archocentrus nigrofasciatus in relation to temperature

In order to conserve and culture the cichlid fish Archocentrus nigrofasciatus, more information about its reproductive biology and its larval behavior and morphogenesis is necessary. Currently, temperatures ranging from 21 to 27 °C are used in ornamental aquaculture hatcheries. Lower temperatures are preferred to reduce the costs of water heating, and 23 °C is usually the selected temperature. However, there is limited information on culturing protocols for ornamental species and most of the information generated on this topic remains scarce. Thus, the present study examines the morphological development of Archocentrus nigrofasciatus during the yolk‐sac period up to the age of 100 h post‐hatching in relation to 2 temperature regimes used in ornamental aquaculture: a temperature of 27 °C (thermal optimum) and a decreased temperature of 23 °C (thermal tolerance). The results of this study suggest that the 27 °C temperature generates intense morphological changes in yolk‐sac development in a shorter period. This has advantages as it reduces the time of yolk‐sac larval development, and, thus, minimizes the transition phase to exogenous feeding and maximizes the efficiency at which yolk is converted into body tissues. The present paper provides necessary information to produce freshwater ornamental fish with better practices so as to increase larval survival and capitalize on time for growth.     

Influence of Dietary Lipid and Temperature on Bactericidal Activity of Channel Catfish Macrophages

Abstract

We fed juvenile channel catfish Ictalurus punctatus purified diets containing different lipid sources for 90–110 d to examine the influence of dietary lipids on intracellular killing of Edwardsiella ictaluri by pronephros macrophages. The diets contained either menhaden oil, soybean oil, beef tallow, or a combination ofall three lipids. We performed the study with nonimmune and immunized fish acclimated to optimal (28°C) and suboptimal (19°C) temperatures. In both temperature trials, bactericidal activity was positively correlated with the level of n-3 fatty acids in the diet (particularly long-chain highly unsaturated fatty acids). Vaccination resulted in substantially enhanced bactericidal activity, except at suboptimal temperature in fish fed the diet containing beef tallow (primarily saturated and monoenoic fatty acids). Overall results were similar at optimal and suboptimal temperatures. This study demonstrates the potential for nutritional manipulation of disease resistance in fish and illustrates the need for standardized diets in fish health research.     

Metabolism and thermoregulation of individual and clustered long-fingered bats,Miniopterus schreibersii,and the implications for roosting

Oxygen consumption of individual long-fingered bats, Miniopterus schreibersii, was measured at air temperatures (T_r) between 2 and 42°C and that of clusters of four and six bats between 5 and 30°C. BMR of individuals was estimated to be 2.29 ml O₂ g ¹ h^-1 between 34 and about 38°C. M. schreibersii showed two different responses to T_a, either maintaining body temperature (T_b) by increasing metabolic rate, or allowing T_b to fall close to ambient temperature and conserving energy (torpor). Euthermic clusters of four and six bats had lower rates of mass-specific oxygen consumption and lower thermal conductances than individuals at equivalent temperatures and torpid clusters maintained a greater temperature differential between T_b and T_a. Lowest rates of metabolism were measured for euthermic bats at air temperatures higher than those available to them in their natural roost in summer, so typical roost temperatures result in metabolic rates that are about 2 4–3.1 times estimated BMR. Roost temperatures of M. schreibersii in South Africa are substantially higher in winter than are generally accepted as being suitable for hibernation but these do not substantially affect torpid metabolic rates, which are low and independent of ambient temperature below 22°C. Clearly at least some species of insectivorous bats are capable of hibernating at temperatures generally considered to be too warm for this activity.     

Influence of Temperature on the Arginine and Methionine Requirements of Young Broiler Chicks

Two experiments were conducted with Ross × Ross broiler chicks in battery brooders from 7 to 21 d of age to determine the Arg and Met requirements of young broiler chicks at control (25°C) and warm (35°C) temperatures. In both experiments, 1-d-old broiler chicks were fed a corn and soybean meal based starter diet for 7 d. Six replications (2 replicates in each of 3 rooms per temperature) with 8 chicks each were used for each treatment. In experiment 1, the basal diet was based on corn (34.52%), whey (26.96%), corn gluten meal (16.53%), soybean meal (11.74%), and poultry fat (23% of CP and 3.20 kcal/g of ME_n). Six levels of Arg (0, 0.1, 0.2, 0.3, 0.4, and 0.5%), supplementing the basal diet containing 0.95% Arg, were the dietary treatments. A broken-line linear model was used to estimate chick Arg requirements. Based on body gain and feed conversion ratio (FCR) data, respectively, the Arg requirements of young Ross × Ross broiler chicks raised at 35°C were 1.15 ± 0.03% and 1.13 ± 0.02%, whereas those of chicks at 25°C were 1.26 ± 0.03% and 1.27 ± 0.02%. In experiment 2, the influences of temperature and dietary Arg on the Met requirements of young broiler chicks were investigated. The basal diet was based on corn (53.45%), soybean meal (37.72%), and poultry fat (23% of CP and 3.20 kcal/g of ME). Experiment 2 had a 6 × 2 factorial arrangement, with the basal diet (0.35% Met and 1.52% Arg) supplemented with 6 levels of dietary Met (0, 0.05, 0.1, 0.15, 0.2, or 0.3%) and 2 levels of dietary Arg (0 and 1.0%). When chicks were fed a corn-soybean meal basal diet containing 1.52% Arg, the Met requirements of young Ross × Ross broiler chicks raised at 35°C were 0.43 ± 0.02% and 0.43 ± 0.03%, whereas those of chicks at 25°C was 0.43 ± 0.01% and 0.48 ± 0.03%, based on body gain and FCR data, respectively. When Arg levels were increased to 2.52%, the Met requirement of young Ross × Ross broiler chicks was greater at both temperatures (P < 0.05). The requirements of chicks raised at 35°C were 0.50 ± 0.02% and 0.49 ± 0.02% and at 25°C were 0.59 ± 0.03% and 0.57 ± 0.02%, based on body gain and FCR data, respectively. Temperature and amino acid balance may both affect the amino acid requirements of broilers.     

Effects of Hydrogen Peroxide (H2O2) on Fresh and Cryopreserved Buffalo Sperm Functions During Incubation at 37°C In Vitro

The magnitude of damage to buffalo spermatozoa during incubation with different levels of H₂O₂ was assessed. A total number of 24 ejaculates from four Murrah buffalo bulls were analysed in the study. Each ejaculate was split into two parts (part I and II). Part I was extended in Tris–egg yolk–citrate extender (20% egg yolk:7% glycerol), equilibrated (4 h at 5°C) and cryopreserved in 0.5‐ml French straws and stored in liquid nitrogen. The other part was utilized for fresh semen studies. The sperm in fresh, equilibrated and frozen–thawed semen was separated by centrifugation (1500 g ; 15 min) and were washed with sperm TALP. The sperm cells were re‐suspended in incubation TALP at the rate of 10⁸ sperm cells per millilitre and incubated with 0, 10, 25, and 50 μm H₂O₂ per ml at 37°C. Sperm motility, viability and intact acrosome percentages were assessed at 15‐min intervals up to 60 min of incubation. Lipid peroxidation levels of sperm were assessed at 0 and 60 min of incubation. The results of the experiment revealed that sperm motility decreased drastically during incubation with H₂O₂. Among the different levels of H₂O₂, the 50‐μm H₂O₂‐incorporated group had significantly (p < 0.05) higher malonaldehyde (MDA) level than the other groups. In the 50‐μm H₂O₂‐incorporated group, the MDA levels in fresh, equilibrated and frozen–thawed semen after incubation for 60 min were 961.6 ± 12.7, 991.8 ± 10.3 and 1234.9 ± 9.6 nm per 10⁹ spermatozoa respectively. An inverse relationship was observed between sperm motility, viability, intact acrosome percentages and concentration of H₂O₂ and duration of incubation. The decrease in sperm functions with duration of incubation and concentration of H₂O₂ was significantly (p < 0.05) higher in frozen–thawed than fresh and equilibrated spermatozoa.     

Evaluation of Emamectin Benzoate for the Control of Experimentally Induced Infestations of Argulus sp. in Goldfish and Koi Carp

Abstract

The effect of 0.2% emamectin benzoate (SLICE; Intervet/ Schering-Plough Animal Health, Roseland, New Jersey) administered in top-dressed, pelleted commercial fish feed was evaluated for control of freshwater Argulus sp. in goldfish Carassius auratus and koi carp, a variant of common carp Cyprinus carpio, in freshwater aquaria at 24–25°C. Sixteen individually housed goldfish were each exposed to 37 Argulus. The number of fish lice attached to each fish at the start of the experiment was not determined; however, the total number of motile fish lice in each aquarium (on fish and in the water) was determined at the start and end of each experiment. Eight goldfish were fed the control diet (0 μg · kg fish biomass^?1 · d^?1) and eight were fed the medicated diet (50 μg · kg fish biomass^?1 · d^?1) for seven consecutive days. After treatment, fish louse infestation in controls was 20.5 ± 1.5 (mean ± SE) lice per fish. No Argulus were found on fish in the treated group. In a separate experiment, 10 individually housed koi were each exposed to 128 Argulus. Five koi were fed the control diet and five were fed a low-dose medicated diet (5 μg · kg fish biomass^?1 · d^?1) for 7 d. After treatment, fish louse infestation among the controls was 14.6 ± 3.8 lice per koi. No Argulus were found on koi in the treated group. Hence, a 7-d regimen of oral emamectin benzoate controlled experimental infestation of Argulus when administered to goldfish at 50 μg · kg fish biomass^?1 · d^?1 and to koi at 5 μg · kg fish biomass^?1 · d^?1.

Received March 24, 2010; accepted January 17, 2011     

Pharmacokinetics of florfenicol in blunt‐snout bream (Megalobrama amblycephala) at two water temperatures with single‐dose oral administration

The pharmacokinetics and residue elimination of florfenicol (FFC) and its metabolite florfenicol amine (FFA) were studied in healthy blunt‐snout bream (Megalobrama amblycephala, 50 ± 10 g). The study was conducted with a single‐dose (25 mg/kg) oral administration at a water temperature of 18 or 28°C, while in the residue elimination study, fish were administered at 25 mg/kg daily for three consecutive days by oral gavage to determine the withdrawal period (WDT) at 28°C. The FFC and FFA levels in plasma and tissues (liver, kidneys and muscle) were analysed using high‐performance liquid chromatography (HPLC). A no‐compartment model was used to analyse the concentration versus time data of M. amblycephala. In the two groups at 18 and 28°C, the maximum plasma concentration (C_max) of FFC was 5.89 and 6.21 μg/ml, while the time to reach C_max (T_max) was 5.97 and 2.84 hr, respectively. These suggested that higher temperature absorbed more drug and more quickly at M. amblycephala. And the elimination half‐life (T_1/2kβ) of FFC was calculated as 26.75 and 16.14 hr, while the total body clearance (CL) was 0.09 and 0.15 L kg^?1 hr^?1, and the areas under the concentration–time curves (AUCs) were 265.87 and 163.31 μg hr/ml, respectively. The difference demonstrated that the elimination rate of FFC in M. amblycephala at 28°C was more quickly than that at 18°C. The results of FFA showed the same trend in tissues of M. amblycephala. After multiple oral doses (25 mg/kg daily for 3 days), the k (eliminate rate constant) of FFA in M. amblycephala muscle was 0.017, the C₀ (initial concentration) was 3.07 mg/kg, and the WDT was 10 days (water temperature 28°C).