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.     

Carbonic Anhydrase Activities from the Rainbow Trout Lens Correspond to the Development of Acute Gas Bubble Disease

Abstract

Dissolved gas supersaturation is hazardous to fish and can result in gas bubble disease (GBD). Signs of GBD typically include bubbles in the eyes, fins, skin, lateral line, and gill filaments. Ocular abnormalities in diseased salmonids typically occur after aberrant gas production in the eyes. In this study, freshwater rainbow trout Oncorhynchus mykiss were exposed experimentally to percent total gas pressure (TGP%) levels of 104% (control) and 115%. No mortalities occurred during the 7-d experimental period. Effects of GBD were observed externally as a darkened skin, exophthalmia, localized hemorrhage in the eye, and gas bubbles on the operculum. Additional signs included increased swimming activity and, more frequently, panic episodes. Carbonic anhydrase (CA) enzyme activities from the lens and retina were determined at days 0, 1, 3, 5, and 7 of the study. Venous blood gases were also measured on day 7. Retinal pH did not differ between normal and affected fish, but blood characteristics such as the partial pressure of O₂, partial pressure of CO₂, carboxyhemoglobin level, and bicarbonate ion concentration were significantly elevated in affected fish relative to normal fish. Venous blood pH and oxyhemoglobin levels were not significantly different between affected and normal fish. Patterns of response to total dissolved gas levels differed between the lens and the retina. Mean CA activities in the lenses of fish exposed to a TGP% level of 115% were significantly below those of control fish. However, retinal CA activities did not significantly differ between the two groups over the course of the experiment. These findings show that dissolved gas supersaturation reduces CA activity in the rainbow trout lens.

Received June 17, 2010; accepted March 20, 2011     

Carbonic anhydrase activities from the rainbow trout lens correspond to the development of acute gas bubble disease

Dissolved gas supersaturation is hazardous to fish and can result in gas bubble disease (GBD). Signs of GBD typically include bubbles in the eyes, fins, skin, lateral line, and gill filaments. Ocular abnormalities in diseased salmonids typically occur after aberrant gas production in the eyes. In this study, freshwater rainbow trout Oncorhynchus mykiss were exposed experimentally to percent total gas pressure (TGP%) levels of 104% (control) and 115%. No mortalities occurred during the 7-d experimental period. Effects of GBD were observed externally as a darkened skin, exophthalmia, localized hemorrhage in the eye, and gas bubbles on the operculum. Additional signs included increased swimming activity and, more frequently, panic episodes. Carbonic anhydrase (CA) enzyme activities from the lens and retina were determined at days 0, 1, 3, 5, and 7 of the study. Venous blood gases were also measured on day 7. Retinal pH did not differ between normal and affected fish, but blood characteristics such as the partial pressure of O2, partial pressure of CO2, carboxyhemoglobin level, and bicarbonate ion concentration were significantly elevated in affected fish relative to normal fish. Venous blood pH and oxyhemoglobin levels were not significantly different between affected and normal fish. Patterns of response to total dissolved gas levels differed between the lens and the retina. Mean CA activities in the lenses of fish exposed to a TGP% level of 115% were significantly below those of control fish. However, retinal CA activities did not significantly differ between the two groups over the course of the experiment. These findings show that dissolved gas supersaturation reduces CA activity in the rainbow trout lens.     

Assessing the Suitability of a Partial Water Reuse System for Rearing Juvenile Chinook Salmon for Stocking in Washington State

Abstract

To assess the suitability of water reuse technology for raising Pacific salmon Oncorhynchus spp. for stocking purposes, fish health and welfare were compared between two groups of juvenile Chinook salmon O. tshawytscha from the same spawn: one group was reared in a pilot partial water reuse system (circular tanks), and the other group was reared in a flow-through raceway. This observational study was carried out over a 21-week period in Washington State. Reuse and raceway fish were sampled repeatedly for pathogen screening and histopathology; fin erosion and whole-blood characteristics were also evaluated. By the study's end, no listed pathogens were isolated from either cohort, and survival was 99.3% and 99.0% in the reuse and raceway groups, respectively. Condition factor was 1.28 in raceway fish and 1.14 in reuse fish; this difference may have been attributable to occasional differences in feeding rates between the cohorts. Fin indices (i.e., length of the longest dorsal or caudal fin ray, standardized by fork length) were lower in reuse fish than in raceway fish, but fin erosion was not grossly apparent in either cohort. The most consistent histological lesion was gill epithelial hypertrophy in reuse fish; however, blood analyses did not suggest any corresponding physiological imbalances. Overall, results suggest that water reuse technology can be employed in rearing juvenile anadromous salmonids for stocking purposes.

Received December 3, 2009; accepted February 14, 2011     

Induction and Evaluation of Proliferative Gill Disease in Channel Catfish Fingerlings

Abstract

Proliferative gill disease (PGD) was first reported in channel catfish Ictalurus punctatus at commercial farms in 1981 and is caused by the myxozoan parasite Henneguya ictaluri. The disease affects the gills and is characterized by severe branchial inflammation, epithelial hyperplasia, lamellar fusion, and lysis of chondrocytes. Presumptive diagnosis is based on the presence of lytic areas in the cartilage of the primary lamellae on microscopic examination and is confirmed histologically by the presence of the organism. In these trials, PGD was induced by exposing channel catfish fingerlings to fresh or aged infectious water collected from a pond containing fish diagnosed with severe PGD. The severity of disease was graded by histological scoring and microscopic examination of wet mounts to determine the percentage of gill filaments containing chondrolytic lesions. Exposure of fish to infectious pond water was shown to produce pathological lesions consistent with PGD, and the percentage of gill filaments containing chondrolytic lesions was positively correlated with histological scoring of gill pathology. The number of trophozoite stages in the gills was shown to increase with the severity of the disease. In most cases, however, parasitic cells were not observed in tissue samples with chondrolytic lesions during the early stages of infection. These observations indicate that pathology and lysis of chondrocytes can occur prior to detection of the organism by histopathology. Exposing fish to infectious pond water that was aged for 1 d produced negligible gill pathology and implies that the infectivity of the H. ictaluri actinospore stage is short lived. Removing fish from the source of infection promoted repair of damaged gill tissue; within 14 d of fish transfer to clean water, gill pathology associated with the acute infection was negligible.     

Prevalence of Renibacterium salmoninarum among DownstreamMigrating Salmonids in the Columbia River

Abstract

Bacterial kidney disease (BKD) is an important contributor to mortality of salmonids in hatcheries in the Columbia River basin. However, the impact of BKD on the survival of downstream migrants is difficult to determine because there is little information on the disease-related mortality among these fish. In this study, the impact of BKD on juvenile salmonids was examined by determining the percentage of downriver migrants infected with Renibacterium salmoninarum (the causative agent of BKD) and evaluating the effects of salt water on the progress of the disease. During the 2 years of this study, approximately 20% of the three species of migrating hatchery and wild salmonids (Oncorhynchus spp.) collected were infected with R. salmoninarum. Mortality caused by BKD increased when fish were held in salt water.     

Aspects of the pathology of Lamproglena clariae (Copepoda: Lernaeidae) on gills of Clarias gariepinus from the Vaal River system,South Africa

Lamproglena clariae attaches to gill filaments of Clarias gariepinus, penetrates the gill tissue and consumes blood. Hence, the purpose of this study was to investigate the mechanical damage caused by this parasite on gill filaments of the host. Infestation intensity and haematocrit values of fish blood were determined and correlated. Fish were killed, gills dissected out and observed with a dissection microscope. Gill filaments with adult females in situ were fixed in a solution of alcohol, formaldehyde and acetic acid and preserved in 70% ethanol. Fixed specimens were studied by histological examination following staining with Heidenhein’s azan solution. Gross morphology revealed varying degrees of swelling of gill filaments near the attached specimens of L. clariae. Epithelial hyperplasia, resulting in fusion of gill lamellae and necrosis of the host tissue in the vicinity of the parasite were revealed through histological examination. No correlation was evident between the infestation intensity and fish haematocrit values. Although results showed that L. clariae causes localized infestation, high intensity of infestation may be detrimental to the fish. Hyperplasia of epithelial tissue resulting in fusion of lamellae would result in restriction of air passages and thus hinder the process of respiration of the host.     

Assessing the suitability of a partial water reuse system for rearing juvenile Chinook salmon for stocking in Washington State

To assess the suitability of water reuse technology for raising Pacific salmon Oncorhynchus spp. for stocking purposes, fish health and welfare were compared between two groups of juvenile Chinook salmon O. tshawytscha from the same spawn: one group was reared in a pilot partial water reuse system (circular tanks), and the other group was reared in a flow-through raceway. This observational study was carried out over a 21-week period in Washington State. Reuse and raceway fish were sampled repeatedly for pathogen screening and histopathology; fin erosion and whole-blood characteristics were also evaluated. By the study's end, no listed pathogens were isolated from either cohort, and survival was 99.3% and 99.0% in the reuse and raceway groups, respectively. Condition factor was 1.28 in raceway fish and 1.14 in reuse fish; this difference may have been attributable to occasional differences in feeding rates between the cohorts. Fin indices (i.e., length of the longest dorsal or caudal fin ray, standardized by fork length) were lower in reuse fish than in raceway fish, but fin erosion was not grossly apparent in either cohort. The most consistent histological lesion was gill epithelial hypertrophy in reuse fish; however, blood analyses did not suggest any corresponding physiological imbalances. Overall, results suggest that water reuse technology can be employed in rearing juvenile anadromous salmonids for stocking purposes.     

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.     

Eutrophication,Ammonia Intoxication,and Infectious Diseases: Interdisciplinary Factors of Mass Mortalities in Cultured Nile Tilapia

The present study was designed to assess the possible causes of the mass mortalities of Nile Tilapia Oreochromis niloticus at El-Behera Governorate, Egypt, in relationship to environmental and microbiotic factors. Water samples were collected from fish farms at different locations and from Lake Edku to analyze water temperature, water pH, salinity, biological oxygen demand, dissolved oxygen, total ammonia nitrogen, and un-ionized ammonia. A number of moribund and freshly dead fish were sampled and submitted to our laboratory for microbiological, molecular, and histopathological examination. Water analysis of the fish farms revealed noticeable increases in the previously mentioned physicochemical parameters. Clinical examinations of moribund fish showed severe gill rot and massive external and internal hemorrhages. Ordinary and molecular laboratory findings confirmed the presence of Branchiomyces sp. in gill tissue and mixed bacterial fish pathogens (Streptococcus agalactiae, Vibrio alginolyticus, V. parahaemolyticus, Pseudomonas anguilliseptica, and P. aeruginosa) in visceral organs. The histopathological and transmission electron microscopic examinations revealed severe necrosis of gill filaments and blockage of branchial blood vessels and lamellar capillaries with Branchiomyces sp. hyphae and spores mixed with different shapes of bacteria. Severe inflammations were detected in liver, kidney, heart, and brain tissues. Ultimately, we can conclude that the syndrome of mass fish kills in this area is a consequence of ecological damage to the aquatic environment, which is mainly related to natural and anthropogenic factors, as well as to the presence of infectious agents.

Received September 30, 2015; accepted April 12, 2016 Published online August 2, 2016     

The Gill Pathogen Dermocystidium salmonis in Oregon Salmonids

Abstract

Intense infections of the gill pathogen Dermocystidium salmonis were associated with mortality of prespawning chinook salmon Oncorhynchus tshawytscha in several Oregon rivers in 1988. The occurrence of the pathogen in returning adult chinook salmon was monitored in several coastal Oregon stocks from 1989 to 1993. Although the prevalence of the pathogen was high in these fish (up to 66.6%), infection intensities were generally low, and no mortality attributable to D. salmonis was observed. In 1988, the pathogen was associated with a lethal epizootic among juvenile chinook salmon smolts at the Trask State Fish Hatchery near Tillamook, Oregon. Histological examination of gills from heavily infected fish revealed hyperplasia of gill epithelium and fusion of gill lamellae. When naturally infected smolts were transferred from fresh to salt water, the most heavily infected fish died within 10 d, and the number of D. salmonis cysts declined and disappeared from previously infected salmon after 21–42 d.     

Ultrastructural characterization of gills in juveniles of the Argentinian Silverside, Odontesthes bonariensis (Valenciennes, 1835) (Teleostei: Atheriniformes)

An ultrastructural study was performed on the gills of juvenile Argentinian silverside, Odontesthes bonariensis. The gills are composed of two sets of four holobranchs and, in turn, each holobranch consists of a gill arch and two rows of caudolaterally projecting branchial filaments. From the dorsal and ventral surfaces of each filament, branchial lamellae radiate out as foldings of the epithelial layer. Gill rakers are present on each of the gill arches, on the anteromedial side of the arch opposite to the filaments. Gill rakers, gill arches and branchial filaments are covered by a stratified epithelium, whereas branchial lamellae essentially consist of a thin epithelial envelope containing capillaries. In the stratified epithelium, mucous cells, rodlet cells, granular cells, pavement epithelial cells and mitochondria-rich cells are identified. The thin epithelium that lines the lamellae comprises two cell types, outer and inner epithelial cells, and the capillary walls on the inside of the epithelial envelope are defined by pillar cells. The ultrastructure of all these cell types is described and our findings are discussed in light of the existing data on fish gill morphology. In the gills of juvenile Argentinian silverside is of particular interest the characteristics showed by mitochondria-rich cells, such as their arrangement in clusters of 2-3 cells and their small and depressed surface in contact with the aquatic milieu, features which strongly resemble those of euryhaline species.     

Clinical and pathological observations on Streptococcus sp. infection on South African trout farms with gas supersaturated water supplies

Infection with a faecal Streptococcus species belonging to Lancefield group D has been a major cause of mortalities on certain trout farms on the escarpment of the Mpumalanga Province in the Republic of South Africa. Evidence is given of a connection between subclinical gas bubble disease and infection with this organism in rainbow trout, Oncorhynchus mykiss. The condition causes serious losses during the summer months when water temperatures rise above 18 degrees C. The larger table-size fish are affected first and present with exophthalmos before developing signs of a typical bacterial septicaemia. The design of the water supply to the fish ponds on these farms allowed supersaturation of the water with atmospheric gas. Vortex formation at water extraction points appears to have been the main cause of air entrainment into the water supply leading to the ponds. By correcting the faults responsible for the development of hyperbaric gas pressure in the water, it was possible to control this disease on two badly affected farms. The overlapping similarities in the clinical signs and pathology of gas bubble disease and that seen in fish suffering from Streptococcus D infection are discussed, as well as the diagnostically significant changes in the gill tissues that are specific to subclinical exposure to gas supersaturation in the water. An explanation is given for the increased susceptibility to infection with this organism.     

Intraocular oxygen tensions in normal and diseased eyes of farmed halibut

We have previously documented ocular abnormalities in farmed halibut and reported that the production of gas-filled posterior segment cysts appears to be central in the pathogenesis of many, if not all of the signs in these eyes. In a number of fish, gas bubbles may be seen in the anterior chamber of the eye, especially after trauma or strenuous exertion associated with handling. A knowledge of the composition of this gas is important in understanding fully the mechanism of gas and cyst production. Here we report investigations into the composition of gas in the globes of normal fish, fish in which intraocular cysts have been documented previously by ultrasonography and fish where post-mortem examination demonstrated both gas- and fluid-filled choroidal cysts. High levels of oxygen were demonstrated in samples of aqueous from all fish but fish with affected eyes had a statistically significantly higher partial pressure of oxygen than did fish with normal eyes.     

Massive Lernaea cyprinacea Infestations Damaging the Gills of Channel Catfish Polycultured with Bighead Carp

Abstract

During June and July 1998, at least three Arkansas fish farms polyculturing bighead carp Hypophthalmichthys nobilis with channel catfish Ictalurus punctatus suffered major losses of catfish associated with massive infestations by the crustacean parasite Lernaea cyprinacea. The channel catfish had few adult Lernaea attached to their skin, but there were 8–50 Lernaea copepodids on the surface of each catfish gill filament. The copepodids were found grazing on the gill tissue, and their feeding activity was associated with gill damage including epithelial hyperplasia, telangiectasis, and hemorrhage. Bighead carp in the same ponds were reported to have had numerous adult Lernaea on their skin but did not die during the epizootic. It is possible that the filter-feeding apparatus of the carp captured the copepodids, thus preventing heavy infestation of the gill filaments. Lernaea copepodids have not been implicated previously in fish losses resulting from parasite damage to the gills. The loss of catfish in these cases is likely to be due to their polyculture with the bighead carp, a species that provides an excellent host for adult Lernaea.     

Rearing unit-level factors associated with bacterial gill disease treatment in two Ontario, Canada government salmonid hatcheries

Early-rearing salmonids in Ontario Ministry of Natural Resources (OMNR) fish hatcheries have been consistently affected by bacterial gill disease (BGD) (causative agent: Flavobacterium branchiophilum) for many years. Separate retrospective epidemiological investigations of BGD treatments at two OMNR fish hatcheries (Hatcheries A and B) for the 1999 production year were conducted using on-site hatchery records. Both investigations were carried out at the rearing unit-level, with early-rearing (<9 months of age) “tank-lot” as the unit of analysis to identify unique fish populations over time. Multivariable repeated measures logistic regression models were created for both hatchery datasets, controlling for lot-level and species effects. For Hatchery A, the species brook trout (Salvelinus fontinalis) and brown trout (Salmo trutta) were significantly associated with BGD treatment, as well as lower water exchange rate, and higher feeding and mortality percentages during the 2 weeks previous to BGD treatment. At Hatchery B, the species brook trout (S. fontinalis) and splake (Salvelinus namaycush × S. fontinalis) were significantly associated with BGD treatment, as well as lower individual fish weights and treatment for BGD during the previous week. These results emphasize the importance of water quality, feeding rate, fish size and prior mortality on the development of BGD. Significant hatchery and species effects were evident, and future observational research on BGD must account for these factors in their design and analysis.     

Pathological Changes Associated with Vitamin C Deficiency in Walleyes

Abstract

Walleyes Stizostedion vitreum were fed three experimental diets containing 0, 96, and 190 mg vitamin C/kg for 20 weeks. Retarded growth, increased mortality, and clinical signs of vitamin C deficiency were seen only in the group fed the vitamin-C-free diet. Skeletal deformities, primarily lordosis, were evident in 76% of this group. Pathological changes associated with vitamin C deficiency in walleyes included twisted cartilage of gill filaments and extreme dislocation of vertebrae. Vertebral lesions included focal hemorrhage, compressed spinal cord, and displacement of adjacent kidney and skeletal muscle. Breaking of the isthmus, a clinical sign not previously described for vitamin-C-deficient fish, was the cause of death in 27% of walleyes fed the vitaminC-free diet. Hemorrhagic fins, skin, and eyes were seen occasionally. A diet that was not supplemented with vitamin C but that contained 96 mg vitamin C/kg was adequate for normal growth and prevention of vitamin-deficiency signs in walleye.     

Gill epithelium of an angler catfish,Chaca chaca (Siluriformes,Chacidae): Enzyme and glycoprotein histochemistry

A series of histochemical techniques have been employed to localize alkaline phosphatase, acid phosphatase, non-specific esterase, catalase and peroxidase; and to visualize and characterize glycoprotein (GPs) moieties in the epithelium of gill arch, gill filaments and secondary lamellae of an angler catfish Chaca chaca. The epithelium of gill arch and gill filament shows strong alkaline phosphatase activity in the deeper layer epithelial cells; strong non-specific esterase activities in the outer layer epithelial cells; and weak acid phosphatase activity throughout the epithelium. The activity of these enzymes in the secondary lamellae is weak. The catalase and peroxidase show strong activities in the blood cells of the secondary lamellae. Various classes of GPs have been identified and characterized in the mucous secretions of the gill epithelium of C. chaca. These include—GPs with oxidizable vicinal diols, GPs with sialic acid residues without O-acyl substitution and GPs with O-sulphate esters. The functional significance of different enzymes in gill epithelium and the GPs in the mucus secreted on the surface has been discussed with the physiology of the gills in relation to the characteristic habit and habitat of the fish.     

Myxobolus infections of common carp (Cyprinus carpio) in Syrian fish farms

During a survey on Myxobolus infection of pond-cultured common carp in Syria three Myxobolus spp. were found. Myxobolus dispar infected the gill arteries, forming large elongated plasmodia in the gill filaments. The plasmodia of M basilamellaris were located in the gill arches at the base of the filaments. Elongated filiform plasmodia of M. encephalicus were found in the blood vessels of the brain. Despite the common occurrence of the above parasites, no disease symptoms were observed in the infected fish specimens. This is the first report on myxosporean infection of fish from Syrian waters.