Effects of Garlic and Ginger Oils on Hematological and Biochemical Variables of Sea Bass Dicentrarchus labrax

Abstract

This study was conducted to investigate the effects of garlic and ginger oils on hematological and biochemical health characteristics of sea bass Dicentrarchus labrax. Fish were exposed to garlic oil (0.01 or 0.02 mL/L), ginger oil (0.01 or 0.02 mL/L), or a combination of the two oils (each oil at a concentration of 0.005 or 0.01 mL/L) for 96 h via bath immersion. Results showed that the red blood cell count, hematocrit (%), hemoglobin (Hb) concentration (g/dL), mean corpuscular volume (μm³), mean corpuscular Hb (pg), and mean corpuscular Hb concentration (%) were not significantly affected by herb oil exposure. However, some changes in biochemical variables were observed. Sea bass exposed to the 0.005-mL/L garlic oil–ginger oil mixture exhibited a significant increase in serum glucose. Serum total protein and albumin levels decreased in sea bass that were exposed to a garlic oil–ginger oil mixture (0.005 or 0.01 mL/L) or to garlic oil at 0.02 mL/L. Serum globulin levels decreased and triglyceride levels increased in sea bass exposed to 0.02-mL/L garlic oil or to the 0.01-mL/L mixture. The serum lipase level decreased and the cholesterol level increased in fish that were exposed to 0.02-mL/L garlic oil. In summary, ginger oil at 0.01–0.02 mL/L can be used without negative effects, while the garlic oil or garlic oil–ginger oil mixture should be applied at a concentration below 0.005 mL/L for bath immersion of sea bass. This is the first study to examine how garlic oil and ginger oil exposure via bath immersion affects the hematological and biochemical status of sea bass.

Received March 19, 2012; accepted July 2, 2012     

A Single Sea Lamprey Attack Causes Acute Anemia and Mortality in Lake Sturgeon

Abstract

The effects of sea lamprey Petromyzon marinus parasitism on hematological variables have not been quantified for lake sturgeon Acipenser fulvescens. Our study objectives were to (1) assess changes in lake sturgeon hematology immediately after a single sea lamprey attack and after a 2-week recovery period and (2) assess changes in the histological condition of major hematopoietic organs. Lake sturgeon from four size-groups (470–570, 570–650, 650–760, and 950–1,500 mm fork length) were individually subjected to a sea lamprey attack in a series of 55 experimental trials. Survival of lake sturgeon after a single sea lamprey attack was size dependent, with fish in smaller size-groups exhibiting higher direct and indirect mortality than individuals in larger size-classes. The most sensitive blood chemistry variable was hematocrit: each 1% decline in hematocrit resulted in a 5.1% increase in mortality risk. Other important variables were plasma protein level, with a 10-g/dL decline resulting in a 4.2% increase in mortality risk; and hemoglobin, with a 1-g/dL decline resulting in a 2.9% increase in mortality risk. Most of the surviving lake sturgeon were unable to restore hemoglobin, hematocrit, and plasma protein to pre-attack levels by the end of the 2-week recovery period. We developed an index of histological spleen condition, which indicated that short-duration (<5-d) sea lamprey attachments depleted red blood cell reserves faster than longer-duration attacks. Our study results indicate that sea lamprey parasitism has the potential to induce acute anemia in lake sturgeon and that nonlethal attacks on smaller (<760-mm) fish can have serious physiological implications.

Received August 3, 2011; accepted February 2, 2012.     

Influence of light colours on growth and stress response of rainbow trout (Oncorhynchus mykiss) under laboratory conditions

Summary The influence of light colours on growth and stress response in rainbow trout Oncorhyncus mykiss (15.16 ± 0.29 cm; 32.27 ± 1.18 g) was studied. Fish were reared in 16 glass aquaria (140 × 30 × 80 cm) each with 12 fish under one of four different lighting spectra: yellow (546 nm), red (605 nm), blue (470 nm) and white (full spectrum, control). Experiments lasted 125 days. The stress response was evaluated by measuring cortisol levels. Body weight and total length of the fish reared under yellow light were greater compared with the other colour regimes while feed conversion ratio significantly lowers. Condition factor and specific growth rate, however, were not differentiated among experimental light treatments. Stressed fish showed lower cortisol levels under yellow light compared with other light exposures. The study indicates that under laboratory conditions, rainbow trout grow best under yellow light and that yellow light lowers the stress‐induced cortisol response in this fish species.     

Effect of Temperature and Dissolved Oxygen Concentration on Edwardsiella ictaluri in Experimentally Infected Channel Catfish

Abstract

Channel catfish Ictalurus punctatus were experimentally infected with Edwardsiella ictaluri by immersion exposure. After clinical disease ran its course for 52 d, the surviving fish were exposed to one of the following environmental regimes in troughs: 25°C with aeration, 25°C with no aeration, or variable temperature (18–23°C) with no aeration. After 29 d of exposure to the environmental regimes, various organs and tissues of the fish were assayed to determine the effects of these conditions on E. ictaluri concentrations (colony-forming units/mL of tissue sample). The concentrations of this pathogen were significantly (P < 0.05) higher in all tissues (trunk kidney, liver, head kidney, blood, spleen, gallbladder, muscle, brain, and gonad) 52 d postinfection than 29 d after exposure to any of the environmental regimes (81 d postinfection). Fish exposed to a near-normal concentration of dissolved oxygen (6.4 mg/L) and a constant temperature of 25°C had E. ictaluri concentrations that were significantly (P < 0.01) lower than those offish exposed to a low oxygen concentration (2.6 or 1.8 mg/L) and either a constant or a variable temperature.     

North American Strain of Viral Hemorrhagic Septicemia Virus is Highly Pathogenic for Laboratory-Reared Pacific Herring

Abstract

Specific-pathogen-free Pacific herring Clupea pallasi were reared in the laboratory from eggs and then challenged at 5, 9, and 13 months of age by waterborne exposure to low (10^1.5–2.5 plaque-forming units [PFU] per milliliter), medium (10^3.5–4.5 PFU/mL), or high (10^5.5–6.5 PFU/mL) levels of a North American isolate of viral hemorrhagic septicemia virus (VHSV). The fish were extremely susceptible to the virus, showing clinical disease, mortality approaching 100%, and only a limited increase in resistance with age. Mortality began 4–6 d after exposure and peaked at approximately day 7 in fish exposed to high levels of virus. Whereas the mean time to death showed a significant dose response (P < 0.001), the percent mortality and virus titers in dead fish were generally high in all groups regardless of initial challenge dose. External signs of disease were usually limited to 1–2-mm hemorrhagic areas on the lower jaw and isthmus and around the eye, but 2 of 130 infected fish exhibited extensive cutaneous hemorrhaging. Histopathologic examination of tissues from moribund fish sampled at 2–8 d after exposure revealed multifocal coagulative necrosis of hepatocytes, diffuse necrosis of interstitial hematopoietic tissues in the kidney, diffuse necrosis of the spleen, epidermis, and subcutis, and occasional necrosis of pancreatic acinar cells. Virus titers in tissues of experimentally infected herring were first detected 48 h after exposure and peaked 6-8 d after exposure at 10^7.7 PFU/g. Fish began shedding virus at 48 h after exposure with titers in the flow-through aquaria reaching 10^2.5 PFU/mL at 4–5 d after exposure, just before peak mortality. When the water flow was turned off for 3 h, titers in the water rose to 10^3.5 PFU/mL, and the amount of virus shed by infected fish (on average, greater than 10^6.5 PFU/h per fish) appeared sufficient to sustain a natural epizootic among schooling herring. Taken together, these data suggest that VHSV could be a significant limiting factor for populations of Pacific herring.     

Efficacy of Hydrogen Peroxide to Control Parasitic Infestations on Hatchery-Reared Fish

Abstract

The efficacy of hydrogen peroxide to control external parasitic infestations on juvenile (10–33-g) rainbow trout Oncorhynchus mykiss was evaluated in three clinical field trials. Fish were exposed to hydrogen peroxide concentrations ranging from 0 to 560 mg/L for 30 min once every other day for a total of three treatments. Pre- and posttreatment skin scrapes and gill wet mounts of test fish were microscopically examined to identify and enumerate external parasites. Infestation severity was classified as nonexistent (0 organisms), low (1–10 organisms), moderate (11–20 organisms), or high (?21 organisms). In trial 1, pretreatment skin examinations revealed a severe infestation of the protozoan Ambiphrya on all fish examined. Posttreatment skin examinations conducted within 24 h of the last treatment indicated that all hydrogen peroxide treatments eliminated Ambiphrya, whereas control fish remained severely infested with the protozoan. In trial 2, pretreatment examinations of skin and gill samples indicated a high infestation of the trematode Gyrodactylus (skin) and the protozoan Trichodina (gills) on all fish. Posttreatment examinations conducted within 24 h of the last treatment indicated that Gyrodactylus was eliminated from the skin of all treated fish; however, the high infestation of Trichodina remained on the gills of the test fish. All control fish had high infestation levels of both parasites. A high infestation of Ambiphrya was found on the skin of test fish before treatment (trial 3). Posttreatment examinations conducted 14 d after the last treatment revealed that 56% of the fish were parasite free, whereas the remaining test fish had low infestation levels. Control fish remained severely infested with the parasite. Based on the efficacy data, all hydrogen peroxide treatment regimens were efficacious in the control of Ambiphrya and Gyrodactylus.     

Experimental Transmission and Pathogenicity of a Viral Erythrocytic Infection in Sea Bass Dicentrarchus labrax

Abstract

The experimental transmission of a viral erythrocytic infection (VEI) in sea bass Dicentrarchus (=Morone) labrax of the Mediterranean is reported. Transmission procedures included the intraperitoneal inoculation of healthy fishes with blood and hematopoietic tissue extracts from VEI-affected fishes, direct contact between VEI-affected and unaffected fishes, and rearing of fishes in unfiltered, nonirradiated seawater that had been used for holding VEI-affected fishes. Successful transmission of the infection was achieved in all experimental conditions, as ascertained by the presence of inclusion bodies in Giemsa-stained blood cells and subsequently confirmed by indirect immunofluorescent staining. Transmission of VEI by the inoculation of filtered cell-free extracts from affected tissues confirmed not only the infectious character of the disease but also its viral etiology. Two different patterns of effects on the hematological variables were observed: an erythroblastic polycythemia occurred 3 months after infection, whereas a moderate anemia appeared after 5–6 months. Fish growth was also negatively affected by VEI. Sea bass infected with VET were found to be more susceptible to bacterial challenge infection with Vibrio anguillarum isolate AO 18 than were control fish; the V. anguillarum dose that was lethal to 50% of the test fish was lower for VEI-affected fish, and their cumulative mortality in the bacterial challenge was higher. The injection of cyclophosphamide produced immunosuppression in unaffected sea bass and an increase in the level of infection in VEI-affected fishes; these results suggest the role of the immunological status of the host sea bass in the development of VEI.     

Influence of Infection with Renibacterium salmoninarum on Susceptibility of Juvenile Spring Chinook Salmon to Gas Bubble Trauma

Abstract

During experiments in our laboratory to assess the progression and severity of gas bubble trauma (GBT) in juvenile spring chinook salmon Oncorhynchus tshawytscha, we had the opportunity to assess the influence of Renibacterium salmoninarum (Rs), the causative agent of bacterial kidney disease, on the susceptibility of salmon to GBT. We exposed fish with an established infection of Rs to 120% total dissolved gas (TDG) for 96 h and monitored severity of GBT signs in the fins and gills, Rs infection level in kidneys by using an enzyme-linked immunosorbent assay (ELISA), and mortality. Mortality occurred rapidly after exposure to 120% TDG, with a LT20 (time necessary to kill 20% of the population) of about 37 h, which is at a minimum about 16% earlier than other bioassays we have conducted using fish that had no apparent signs of disease. Fish that died early (from 31 to 36 h and from 49 to 52 h) had significantly higher infection levels (mean ± SE ELISA absorbance = 1.532 ± 0.108) than fish that survived for 96 h (mean ± SE ELISA absorbance = 0.828 ± 0.137). Fish that died early also had a significantly greater number of gill filaments occluded with bubbles than those that survived 96 h. Conversely, fish that survived for 96 h had a significantly higher median fin severity ranking than those that died early. Our results indicate that fish with moderate to high levels of Rs infection are more vulnerable to the effects of dissolved gas supersaturation (DGS) and die sooner than fish with lower levels of Rs infection. However, there is a substantial amount of individual variation in susceptibility to the apparent cumulative effects of DGS and Rs infection. Collectively, our findings have important implications to programs designed to monitor the prevalence and severity of GBT in juvenile salmonids in areas like the Columbia River basin, and perhaps elsewhere.     

Attempts to Control Flexibacter columnaris Epizootics in Pond-Reared Channel Catfish by Vaccination

Abstract

Evaluation of bacterins for the immunization of channel catfish Ictalurus punctatus against Flexibacter columnaris was carried out over 5 years. Groups of 60,000–100,000 pondreared channel catfish, weighing an average of 1.5–4.1 g, were vaccinated each year with formalininactivated Flexibacter columnaris bacterins by immersion. Bacterins were prepared from isolates of the preceding year's epizootic. Equal numbers of nonvaccinated channel catfish reared under similar conditions served as controls. Fish were vaccinated during May each year when water temperatures ranged from 16 to 18°C. Data were recorded daily from early June through midSeptember as water temperatures rose to 24–26°C. Deaths were recorded daily, and the number of hours required for antibiotic treatments were monitored. Analysis of the data indicated a general trend toward a beneficial effect of vaccination, particularly for larger fish. Mortality and hours of antibiotic treatment were significantly higher for most control groups than for vaccinated groups.     

Preliminary Assessment of Dietary Mannanoligosaccharides on Growth Performance and Health Status of Gilthead Seabream Sparus auratus

Abstract

A feeding trial was performed to assess the potential beneficial effect of two levels of mannanoligosaccarides (MOS) on the growth performance, feed utilization, hematological parameters, and liver histopathology of gilthead seabream Sparus auratus (also known as gilthead bream). Mannanoligosaccarides were added at the rates of 2 and 4 g/kg to a fish-meal-based control diet, and each diet was given (twice daily [midmorning and midafternoon] to apparent satiation) to triplicate groups of gilthead seabream growers (mean weight = approximately 170 g) in sea cages. The trial lasted 12 weeks, and the average ambient water temperature ranged from 19.6°C to 24.7°C during the experimental period. At the end of the experiment, fish attained market size (350–450 g) and their health status was evaluated by blood analysis and liver histology. There were no differences in survival rates among fish fed experimental diets. However, there were significant improvements in both growth and feed utilization among fish fed diets supplemented with MOS. Hemoglobin (Hb) and hematocrit (Ht) levels and erythrocyte, leukocyte, and thrombocyte (Thr) counts were unaffected by any dietary MOS. The levels of Hb (g/dL; mean ± SD) and Ht (%; mean ± SD) were 11.0 ± 2.5 and 45.6 ± 6.7 for the control group, 11.1 ± 1.7 and 39.3 ± 8.0 for the 2-g/kg group, and 11.2 ± 1.9 and 40.2 ± 8.4 for the 4-g/kg group. The mean Thr count ranged from 47.6 to 53.8 × 10³/mm³. Despite the apparently higher Thr counts for fish fed diets supplemented with MOS, these differences were not significant. Moreover, no histopathological differences were observed in liver tissue cross sections between control and treatment groups. These results suggest that supplementation of diets with MOS had no significant effects on general fish health.

Received February 20, 2011; accepted September 12, 2011     

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.     

Communications: Tolerance of Rainbow Trout to Dissolved Oxygen Supplementation and a Yersinia ruckeri Challenge

Abstract

Dissolved oxygen accounted for a significant proportion of the variation in mortality rate of rainbow trout Oncorhynchus mykiss acclimated for 10 weeks to a range of dissolved oxygen levels and challenged with Yersinia ruckeri. Moderate levels of oxygen supersaturation (150%) resulted in greater cumulative mortality (17.9%) among fish exposed to a 0.5-h static bath challenge with Y. ruckeri (1 × 10⁷ colony-forming units/mL). In contrast, hypoxic (70%) and normoxic (100%) levels of dissolved oxygen resulted in 12.8% and 10% cumulative mortality. Higher oxygen levels provided no additional “protection” against infection by Y. ruckeri in this study.     

Fin Condition of Atlantic Salmon Reared at High Densities in Heated Water

Abstract

We assessed the condition of dorsal and pectoral fins of parr of Atlantic salmon Salmo salar that were reared for 70 d at 17.5°C to final densities of 14–55 kg/M² (rearing-unit bottom area) or 80–310 kg/m³ (rearing-unit volume). Pectoral fins were not significantly affected by rearing density, but dorsal fins were more eroded in fish reared at the highest density than in fish checked before the rearing experiments or in those reared at lower densities. Densities as great as 41 kg/m² (229 kg/m³) did not affect fin quality of Atlantic salmon reared at 17.5°C.     

Effects of Dietary Zinc and Calcium on Select Immune Functions of Channel Catfish

Abstract

The effects of dietary zinc and calcium on immunocompetence of channel catfish Ictalurus punctatus (mean initial weight = 3.1 g/fish) were determined after a 10-week feeding trial. Fish were fed daily amounts equal to 3% of their body weight of one of six purified diets that were deficient in zinc (2 mg/kg of diet), calcium (0.02% of diet), or both; replete with zinc (20 mg/kg of diet) and calcium (0.5% of diet); or excessive in zinc (200 mg/kg of diet) or calcium (2.5% of diet). Immunocompetence of immunized and nonimmunized fish fed the various diets was assessed by measuring serum immunoglobulin M levels and mortality after an intraperitoneal injection of live Aeromonas hydrophila. Channel catfish fed diets deficient in zinc, calcium, or both had significantly (P ≤ 0.05) less weight gain and survival as compared with those fed diets replete with or excessive in zinc or calcium. Tissue mineral concentrations of fish indicated that various levels of zinc and calcium nutriture existed when immune functions were tested. Diet affected mortality of only nonimmunized fish in the bacterial-challenge test; zinc deficiency and calcium excess decreased mortality. Whereas growth and survival of fish were reduced by dietary zinc or calcium deficiency, the measured immunological characteristics were not equally impaired by these mineral deficiencies.     

Experimentally Induced Whirling Disease I. Dose Response of Fry and Adults of Rainbow Trout Exposed to the Triactinomyxon Stage of Myxobolus cerebralis

Abstract

The intensity and prevalence of whirling disease was tested by exposure of 2-monthold fry and 1-, 2-, and 3.5-year-old adults of rainbow trout Oncorhynchus mykiss to a known number of laboratory-produced Myxobolus cerebralis at the actinosporean triactinomyxon stage. Fry exposed to graded concentrations of infectivity (triactinomyxons) for 3 h were individually examined for spores of Myxobolus cerebralis 5 and 6 months later. Exposure of fish to the lowest doses, 1 and 10 triactinomyxons per fish, did not result in detectable myxosporean spores. Fish that became lightly infected by a dose of 100 triactinomyxons per fish experienced a decrease in the incidence of infection between 5 and 6 months after exposure. A linear relationship was found between the numbers of recovered myxosporean spores and doses of 100–10,000 triactinomyxons per fish, and the spore burden appeared to plateau at doses of 10,000–100,000 triactinomyxons per fish. Adult fish continuously exposed to the highest dose of triactinomyxons for 3.5 months were infected and asymptomatic, however, the severity of myxosporean infection decreased with increased age of fish. This information may help in controlling whirling disease in salmonids.     

Experimental Transmission of Walleye Dermal Sarcoma to Yellow Perch

Abstract

Walleye dermal sarcoma was transmitted under experimental conditions to yellow perch Perca flavescens. Fish (20 weeks posthatch) were challenged with cell-free tumor filtrates by topical application on the right flank and then held for observation for 25 weeks in 15°C dechlorinated municipal water. Additional treatment groups included yellow perch challenged with cell-free filtrates of walleye discrete epidermal hyperplasia, yellow perch discrete epidermal hyperplasia, normal yellow perch skin (control), and normal walleye skin (control). Walleye dermal sarcoma was first observed on yellow perch at 20 weeks postexposure, at which time the tumors were small (1–2-mm), slightly raised masses on the right flank of the fish. At the end of the 25-week study, walleye dermal sarcoma was grossly observed in 42% (22 of 53) of the remaining fish and was confirmed by microscopic examination in an additional 29% (9 of 31). Gross and microscopic evaluation of fish at 25 weeks postexposure did not reveal development of lesions in any of the other treatment groups. This study extends the host range of experimentally transmitted walleye dermal sarcoma virus to include the yellow perch.     

Stress Protein Expression in Coho Salmon with Bacterial Kidney Disease

Abstract

The expression of stress protein-70 (SP-70), also known as heat shock protein-70 (HSP-70), was measured by enzyme-linked immunosorbent assay in kidney and liver from coho salmon Oncorhynchus kisutch with bacterial kidney disease (BKD) experimentally induced by injection with Renibacterium salmoninarum. Fish with BKD had more SP-70 in both kidney and liver than did the control fish. The SP-70 measured was derived from the host tissue, not from the pathogen. In fish without clinical disease, SP-70 was not significantly elevated. Elevated stress protein (SP) levels in fish with BKD raise the possibility that BKD or other infectious diseases may interfere with the use of SP induction as a marker of environmental stress in fish and lead to statistical artifacts when the prevalence of disease is not considered.     

DNA Vaccination Partially Protects Muskellunge against Viral Hemorrhagic Septicemia Virus (VHSV-IVb)

Abstract

A DNA vaccine containing the glycoprotein (G) gene of the North American viral hemorrhagic septicemia virus (VHSV) genotype IVb was developed to evaluate the immune response of fish following vaccination and evaluate its efficacy in protecting a susceptible species, the Muskellunge Esox masquinongy, against VHSV-IVb challenge. Seven weeks (539 degree-days) following vaccination with 10 μg of either pVHSivb-G or a control plasmid, Muskellunge were challenged by immersion with 10⁵ plaque-forming units (pfu)/mL of VHSV-IVb. Fish vaccinated with pVHSivb-G had a relative percent survival (RPS) of 45%. Vaccinated fish also had significantly lower mean viral titers in tissues (4.2 × 10² pfu/g) and viral prevalence (4%) than fish receiving the plasmid control vaccine (3.3 × 10⁵ pfu/g; 82%). Neutralizing antibodies were detected 28 d (308 degree-days) postchallenge (11 weeks postvaccination) in 100% of Muskellunge vaccinated with pVHSivb-G compared with only 12% of plasmid-control-vaccinated Muskellunge, suggesting robust induction of a secondary, adaptive immune response. In addition, pVHSivb-G–vaccinated Rainbow Trout Oncorhynchus mykiss challenged 7 d (100 degree-days) postvaccination with the heterologous novirhabdovirus, infectious hematopoietic necrosis virus (IHNV), experienced an RPS of 61%, compared to control fish, suggesting induction of an early and transient nonspecific antiviral immune response. This study provides an important starting point for VHSV-IVb vaccine development and useful information about the antiviral immune response elicited by DNA vaccination in a nondomesticated fish species.

Received May 1, 2016; accepted September 1, 2016     

Early Changes in Pigmented Macrophages in Head Kidney of Channel Catfish Infected with Aeromonas hydrophila

Abstract

Channel catfish Ictalurus punctatus with scarified skin were exposed to the bacterial pathogen Aeromonas hydrophila. Systemic infections developed in 80% of the exposed fish, and the remaining exposed fish had infections limited to the cutaneous lesion. Skin of control fish was injured in the same manner as for the exposed fish, but control fish were not exposed to A. hydrophila nor was A. hydrophila recovered from them. None of the fish died during the 3-d experiment, and gross lesions in head kidneys, including changes in the size or relative weight, were not found in infected fish. Macrophage aggregates in head kidney of systemically infected fish increased 68% in mean area, 28% in number, and 111% in total relative volume. Volume of lipofuscin, the predominate pigment found in channel catfish head kidney, in systemically infected fish was about twice that of control fish. Volume of hemosiderin was about four times as great and macrophages containing hemosiderin were about three times as numerous in systemically infected fish as in control fish. The amount of melanin in macrophages did not change in infected fish. Significant differences in macrophage aggregates and pigments were not found between controls and fish with only superficial infections. Microscopic lesions, other than changes in macrophage aggregates and pigments, were not found in the head kidney. Increases in macrophage aggregates, lipofuscin, and hemosiderin in head kidney of channel catfish were useful for quantification of injury caused by a systemic bacterial infection.     

The Lethal Dose of Largemouth Bass Virus in Juvenile Largemouth Bass and the Comparative Susceptibility of Striped Bass

Abstract

Largemouth bass virus (LMBV) is an iridovirus that was isolated from wild adult largemouth bass Micropterus salmoides in the southeastern United States in 1994. Although originally isolated from moribund wild fish, its virulence to juvenile largemouth bass is uncertain. To help clarify this point, two LMBV titrations were made in juvenile largemouth bass. Titers of LMBV in fathead minnow cells were 10^4.8 and 10^5.8 tissue culture infectious doses—50% cytopathic endpoint (TCID50) per milliliter, respectively. Tenfold serial dilutions of LMBV employed in each cell culture titration, injected intraperitoneally (0.1 mL/fish) into largemouth bass produced calculated lethal dose—50% mortality endpoints (LD50s) of 282 (10^2.45) and 288 (10^2.46) infectious doses in two consecutive infectivity trials. Virus yield of assayed infected fish averaged 10^8.5 TCID50/g and 10^7.7 TCID50/g in viscera of moribund and dead fish in the two trials and 10^6.5 TCID50/g in surviving exposed fish 14 d after infection. In a second experiment, largemouth bass had 100% mortality 5 d after injection while virus immersed fish had a significantly (P ≤ 0.005) lower mortality of 17% at 14 d. Similarly treated juvenile striped bass Morone saxatilis suffered 63% mortality after injection and significantly (P ≤ 0.005) lower mortality of 10% after immersion. In a third study of 25 d, 100% of injected largemouth bass died by 5 d after injection, and all of them were virus-positive. Injected striped bass had a significantly (P ≤ 0.005) lower mortality of 24%; all three fish were virus-positive initially, two fish were virus-positive at 18 d, and none were positive at 25 d. Juvenile largemouth bass were highly susceptible to LMBV injection and striped bass were moderately susceptible, but both species were only mildly susceptible when exposed by immersion.