PKX,the Causative Agent of Proliferative Kidney Disease (PKD) in Pacific Salmonid Fishes and Its Affinities with the Myxozoa1

Proliferative kidney disease (PKD), caused by an unclassified protozoan (PKX), is reported from Pacific salmon, Oncorhynchus tshawytscha (Walbaum) and O. kisutch (Walbaum), and steelhead trout, Salmo gairdneri Richardson, held at the Mad River Hatchery in California, USA. The cumulative mortality attributed to the disease was 95, 13, and 18% respectively. The mortalities were greatest at mean water temperatures of 12-14°C during July 1983. The ultrastructure of the PKX organism and its associated pathology during clinical disease in all three species were consistent with those of the parasite in rainbow trout (Salmo gairdneri) as described in European outbreaks. Significant mortalities did not occur after August, at which time the parasite could no longer be detected in the salmon species. The steelhead continued to exhibit parasites in the kidney interstitium and epithelium and lumens of the tubules. Myxosporidan trophozoites and developing spores were also observed in the lumens of the kidney tubules of these fish. Although a mixed infection with another parasite may have occurred, evidence suggests that the myxosporidans are later stages of PKX. They were only observed in fish exposed to water with the infective stage and were particularly prominent in recovering fish. The PKX organism is similar to UBO, an unclassified protozoan of carp suspected to be an early stage of Sphaerospora renicola Dyková & Lom. Both parasites infect the blood and kidney, divide by endogeny, and are released by disintegration of the primary mother cell. The intraluminal myxosporean forms show similarities to Sphaerospora spp. in that they are monosporous and sporoblasts are formed within pseudoplasmodia. It is possible that PKX migrates to the lumen of the kidney tubule and subsequently sporulates. If the myxosporean forms are later stages of PKX, then it would belong to the phlyum Myxozoa.     

Effects of cortisol implants on the PKX myxosporean causing proliferative kidney disease in rainbow trout, Salmo gairdneri

The myxosporean (PKX) that causes proliferative kidney disease in salmonid fishes is found primarily in the kidney interstitium of clinically affected fish, where it invokes a severe inflammatory response. Incomplete spores are observed in the lumen of kidney tubules in recovering fish. PKX-infected rainbow trout, Salmo gairdneri, were immunosuppressed with cortisol implants to determine if a reduction in renal interstitial inflammation would enhance the development of the parasite. Immunosuppression of these fish was indicated by high plasma cortisol levels, chronic mortality due to opportunistic pathogens, and depressed leucocrits when compared to infected fish not receiving cortisol implants. Cortisol-treated PKX-infected fish had 20 times the density of interstitial PKX compared with nontreated fish, but showed less interstitial inflammation. All of the former group examined at 4 wk postimplantation exhibited intraluminal sporogonic forms of PKX, whereas no sporogonic forms were detected in the nontreated fish. Suppression of the inflammatory response clearly enhanced the parasite's ability to reach the sporogonic stage, but did not induce complete development of the spore. Possibly the kidney tubules of rainbow trout do not provide the proper physiological environment for complete sporulation of PKX and a nonsalmonid fish may be the primary host.     

Fish infected with Sphaerospora spp. Thélohan (Myxosporea) from waters enzootic for proliferative kidney disease of salmonids

Sphaerospores were found among three species of fish examined from waters known to be enzootic for proliferative kidney disease (PKD) of salmonids. They were detected in the renal tubules of both hatchery-reared rainbow trout (Salmo gairdneri) exposed to the infectious stage of PKD and in chubs (Gila bicolor) in the headwaters of a hatchery where PKD is enzootic. Sticklebacks (Gasterosteus aculeatus) collected near net pens where Pacific salmon had experienced a PKD epizootic were also found to harbor sphaerospores in the lumen of the kidney tubules. The latter two host species contained developmental stages of a myxosporidan in the blood and in the lumen of the kidney tubules which are similar to those of PKX, the causative agent of PKD in salmonid fish. The sphaerospores observed in the rainbow trout are the first to be observed in this species. The similarity to previously observed developmental stages, rarity, and presence of these sphaerospores in salmonid fish from a hatchery where PKD is enzootic suggest that they are the most mature stage of the PKX myxosporidan yet observed.     

Isolation and characterization of Edwardsiella tarda from fall chinook salmon (Oncorhynchus tshawytscha).

A new bacterial pathogen of chinook salmon (oncorhynchus tshawytscha) was isolated from fish in Oregon's Rogue River. The bacteria are biochemically and serologically related to strains of Edwardsiella tarda. Initially isolated from chinook salmon, the bacteria were also pathogenic for steelhead and rainbow trout (Salmo gairdneri), and channel catfish (Ictalurus punctatus). The 50% lethal doses for chinook salmon, steelhead trout, and channel catfish injected intraperitoneally and maintained in 18 degrees C water were 4.1 x 10(6), 5.6 x 10(6), and 4.0 x 10(5) respectively. When chinook salmon and rainbow trout were injected intraperitoneally and held in 12 degrees C water, the mean lethal doses were 6.4 x 10(7) and 1.7 x 10(6), respectively. The invasiveness of the organism was low in steelhead trout exposed to the bacteria by the waterborne route. The optimum growth temperature of the bacteria in brain heart infusion broth was approximately 35 degrees C. The guanine plus cytosine content of DNA obtained from E. tarda isolated from salmon was 59 mol%.     

Peculiarities of the parasitofauna of Atlantic salmon parr (Salmo salar L.), brown trout (Salmo truttae L.), and char (Salvelinus alpinus L.) in the Utsjoki River system (Northern Finland)

Data on the parasite diversity in Salmonidae fish parr from different parts of the Utsjoki River obtained during 1993-1995 and 2006-2007 are presented. Three fish species, Salmo salar L., S. truttae L., and Salvelinus alpinus L., were examined on the presence of helminthes. Twenty species of salmon parasites were found, the majority of which are the parasites with complicated life cycles. Infusorians C apriniana piscium, myxosporidia Chloromyxum januaricus and Myxobolus neurobius, metacercaria of the genera Diplostomum and Apatemon, and the nematode Raphidascaris acus larvae were the most numerous in salmon parasite fauna. Brown trout had the most number of specific parasite species, whereas char was infested by protozoan parasites only.     

On remembrance of the abdominal pores in rainbow trout, Salmo gairdneri Richardson, and some other salmonid spp.

Direct connections (abdominal pores) between the peritoneal cavity and the external environment of elasmobranch fishes and some teleost species including the Salmonidae were recognized almost 100 years ago but over this period their existence in these teleost species has been omitted from anatomical texts. In this report, the abdominal pores of rainbow trout, Salmo gairdneri , Atlantic salmon, Salmo salar , and the cisco, Coregonus artedii , were examined in relationship to ip injected particulate material. Both carbon particles and suspensions of the bacterial kidney disease organism were found to be extruded within masses of macrophages and as free particles through the abdominal pores in fish injected ip 72 h previously. The pores were patent in male and female, mature and immature fish. The role and significance of the abdominal pores in rainbow trout in the clearance from the body of material, including material likely to be used for vaccinating fish, is discussed.     

An electrophoretic analysis of plasma proteins from juvenile Oncorhynchus tshawytscha (Walbaum)

The plasma proteins of juvenile spring chinook salmon, Oncorhynchus tshawytscha , were examined as a function of development. Two-way gel electroprioresis of plasma proteins was conducted on samples taken from fish before initiation of downstream migration, from fish classified as actively migrating and from fish that were prevented from migrating. Analysis of the samples indicated that plasma proteins of fish changed depending on the migratory status of the fish. The results were consistent with the notion that some of the factors normally associated with smoltification in species such as steelhead trout, Salmo gairdneri , and coho salmon, O. kisutch , may well be temporally separated in Idaho O. ishawytscha.     

Comparative sensitivities of diagnostic procedures used to detect bacterial kidney disease in salmonid fishes

Kidney and spleen homogenates from each of 60 coho salmon (Oncorhynchus kisutch) and steelhead trout (Salmo gairdneri) were examined for detection of Renibacterium salmoninarum. The proportions of positives differed widely with the detection procedures used: in coho salmon, 5% were positive by the Gram-stain procedure, 10% by the direct fluorescent antibody test, 48% by bacteriological isolation, 65% by staphylococcal coagglutination, and 73% by counterimmunoelectrophoresis; in steelhead trout, 3% were positive by Gram-stain, 8.3% by fluorescent antibody, 17% by bacteriological isolation, and 67% by counterimmunoelectrophoresis. Renibacterium salmoninarum was not detected in either coho salmon or steelhead trout by immunodiffusion analysis.     

The life history of Plagioporus shawi (McIntosh 1939) (Trematoda:Opecoelida), an intestinal parasite of salmonid fishes.

Podocotyle shawi is transferred to the genus Plagioporus. The miracidium develops and hatches in 11 to 14 days at 22 to 25 C. The operculate snail, Lithoglyphus virens, serves as first intermediate host. Mother and daughter sporocysts develop, the latter producing cotylomicrocercous cercariae within 90 to 130 days. Cercariae encyst in the amphipod, Hyalella azteca; stonefly nymphs of the genus Arcynopteryx; mayfly nymphs of the genera Heptagenia and Paraleptophlebia; chironomid larvae of the genera Chironomus, Polypedilum, Phaenopsectra, Ablabesmyia, and Psectrocladius and caddis larvae of the genera Brachycentrus, Limnephilus, and Hydropsyche. Metacercariae 60 to 100 days old were fed to young steelhead trout, Salmo gairdneri, and partially developed specimens of P. shawi were recovered 26 days later. Stonefly nymphs are believed to be the most important second intermediate host in the natural environment. The parasite seems to be restricted to the Pacific Northwest of the United States and has been reported from the silver salmon, Oncorhynchus kisutch; kokanee, O. nerka; cutthroat trout, Salmo clarki; steelhead trout, S. gairdneri; and the mountain whitefish, Prosopium williamsoni.     

Fish Infected with Sphaerospora spp. Thélohan (Myxosporea) from Waters Enzootic for Proliferative Kidney Disease of Salmonids1

ABSTRACT. Sphaerospores were found among three species of fish examined from waters known to be enzootic for proliferative kidney disease (PKD) of salmonids. They were detected in the renal tubules of both hatchery-reared rainbow trout (Salmo gairdneri) exposed to the infectious stage of PKD and in chubs (Gila bicolor) in the headwaters of a hatchery where PKD is enzootic. Sticklebacks (Gasterosteus aculeatus) collected near net pens where Pacific salmon had experienced a PKD epizootic were also found to harbor sphaerospores in the lumen of the kidney tubules. The latter two host species contained developmental stages of a myxosporidan in the blood and in the lumen of the kidney tubules which are similar to those of PKX, the causative agent of PKD in salmonid fish. The sphaerospores observed in the rainbow trout are the first to be observed in this species. The similarity to previously observed developmental stages, rarity, and presence of these sphaerospores in salmonid fish from a hatchery where PKD is enzootic suggest that they are the most mature stage of the PKX myxosporidan yet observed.     

Parasite Infections in Danish Trout Farms

Samples from 5 Danish freshwater trout farms rearing rainbow trout (Oncorhynchus my kiss) were examined for parasite infections from October 1993 until November 1994 and recorded parasites are listed. In addition, results from an examination of a mariculture net cage system are presented as well. A total of 10 metazoan and 10 protozoan parasites were recorded. The metazoans included Gyrodactylus derjavini, Gyrodactylus salaris, Eubothrium crassum, Triaenophorus nodulosus, Proteocephalus sp., Diplostomum spathaceum, Tylodelphus clavata and Argulus foliaceus from the freshwater farms. The protozoans Hexamita salmonis, Ichthyobodo necator, Ichthyophthirius multifiliis, Apiosoma sp., Epistylis sp., Trichodina nigra, T. mutabilis, T. fultoni, Trichodinella epizootica, and an Ichthyophonus like intestinal parasite were also detected in the freshwater trout farms. Based on lectin binding studies, few fish were found positive for the myxosporean parasite PKX although no clinical cases were reported. In the mariculture system, Lepeophtheirus salmonis and Caligus elongatus were found.     

Some potentials and limits of the leucocrit test as a fish health assessment method

The sensitivity of the leucocrit as a stress tolerance and fish health assessment method was evaluated by subjecting juvenile coho salmon, Oncorhynchus kisutch , or steelhead trout, Salmo gairdneri , to standardized crowding, handling, temperature and disease challenges. The leucocrit was a sensitive indicator of the physiological stress resulting from crowding at population densities of 0·2–0·4 kg l⁻¹, and to the stress of handling and to temperature changes. It was relatively insensitive to physiological sampling procedures which supports its continued development as a stress assessment method.
In the case of fish diseases, subclinical or active Renibacterium salmoninarum and Yersinia ruckeri infections had essentially no effect on leucocrit values. In contrast, active Aeromonas salmonicida infections significantly depressed the leucocrit. However, no change was seen during the subclinical (incubation) phase prior to the development of an epizootic. Thus, the potential of the leucocrit as a fish health assessment method appears limited.     

Serum agglutinating titres against Renibacterium salmoninarum the causative agent of bacterial kidney disease, in rainbow trout, Salmo gairdneri Richardson, and Atlantic salmon, Salmo salar L.

Serum agglutinating titres against Renibacterium salmoninarum were determined from clinically infected, sub-clinically infected and non-infected rainbow trout, Salmo gairdneri Richardson, and Atlantic salmon, Salmo salar L. Titres ≥16 occurred in clinically infected rainbow trout, but declined after a few months when disease signs were absent. Non-infected rainbow trout had titres ≤ 16. No correlation between the agglutinating titre and the level of infection was found in farmed Atlantic salmon. Variable titres were found in wild salmon broodstock although there was no evidence of BKD during 4 years of testing.     

Evidence that infectious stages of Tetracapsula bryosalmonae for rainbow trout Oncorhynchus mykiss are present throughout the year

Proliferative kidney disease (PKD) is a hyperplastic condition of the lymphoid tissue of salmonids infected with the spores of Tetracapsula bryosalmonae, a myxozoan parasite formerly designated PKX, which has recently been described as a parasite of several species of bryozoans. The occurrence of PKD is generally associated with seasonal increase in water temperature, with research indicating that transmission of the disease does not occur below 12 to 13 degrees C. This suggested that the infectious stages are absent from about November to March/April. Here we document the transmission of PKD at water temperatures and seasons previously considered to be non permissive for PKD infection. The exposure of naive rainbow trout Oncorhynchus mykiss (Walbaum) to PKD-infected water ranging from 8 to 13 degrees C during the Autumn, Winter and early Spring, resulted in the infection of kidney interstitium once the trout were transferred to 16 degrees C. In addition, cohabitation studies were conducted with the bryozoan host Fredericella sultana collected from a river at times of low seasonal temperatures because this bryozoan species overwinters as living colonies. Cohabitation of trout with colonies of F sultana in parasite-free city water at 16 degrees C, also led to renal lymphoid tissue infection with the parasite and even to nephromegaly. Our results provide evidence that the infectious stages of T bryosalmonae for rainbow trout were present in the water throughout the entire year and that the impact of temperature on the development of PKD is primarily a result of the kinetics of Tetracapsula multiplication in bryozoan and fish hosts.     

Cells with radiating vesicles in different fish species

A study was made of the ultrastructure of cells with radiating vesicles from the head kidney of different fish species: Atlantic salmon Salmo salar L., carp Cyprinus carpio L., roach Rutilus rutilus (L.), bream Abramis brama (L.), high-body pickarel Spicara staris (L.), long-striped wrasse Symphodus tinca (L.), and from spleen of the rainbow trout Salmo gairdneri Richardson, and the tilapia Sarotherodon mossambicus Peters. A disposition of vesicles around centrioles was characteristics of all the investigated cells. Some differences were marked in the fine structure of vesicles.     

The importance of fish stocking in the dissemination of parasites throughout a group of reservoirs

In view of the known importance of anthropochore movements of fish in disseminating their parasites, it was hypothesized that local movements of fish between neighbouring water bodies for stocking and management purposes would similarly disseminate their parasites, resulting in a high degree of similarity in the parasite communities of fish in water bodies that have been linked by fish transfers. This hypothesis was tested by comparing the helminth parasite communities in brown trout, Salmo trutta Linnaeus and rainbow trout, Oncorhynchus mykiss (Walbaum), in reservoirs in south west England. In one group of six reservoirs there is a recent history of transfers of rainbow trout, and in the second group of four there have been no transfers over the last decade. There was no difference in mean similarity, using Jaccard's Coefficient and a Percentage Similarity Index, between the helminth faunas of trout in the reservoirs linked by transfers and those not so linked, and the distribution of several individual species of helminths was not consistent with their having been disseminated in the course of fish stockings. The hypothesis is thus rejected and it is concluded that anthropochore transfers of trout between reservoirs have been of little or no significance in the dispersal of parasites throughout the reservoirs or in the transfer of helminths from one reservoir to another. The reasons for this unexpected conclusion are discussed in the light of indications that factors unrelated to fish transfers were important determinants of helminth community composition in individual reservoirs.     

Changes in haematocrit values in blood samples treated with and without oxygen: a comparative study with four salmonid species

Changes in haematocrit values under lowered oxygen tension and aerobically treated blood samples of rainbow trout Salmo gairdneri (Richardson), landlocked Baltic salmon S. salar (L.), brown trout S. trutta lacustris (L.) and lake trout Salvelinus namaycush (Walbaum) have been studied in vitro . The mean haematocrit value increased during 2 h incubation under lowered oxygen tension by 32.7 ± 3.1% in rainbow trout, 28.2 ± 2.8% in landlocked Baltic salmon, 29.2 ± 5.6% in brown trout and 25.2 ± 2.8% in lake trout. During corresponding incubation with oxygen the mean haematocrit value decreased below the starting level by 18.1 ± 1.9% in rainbow trout, 18.3 ± 4.8% in landlocked Baltic salmon, 22.4 ± 0.7% in brown trout and 11.8 ± 1.7% in lake trout. Consequently, the changes in the haematocrit values were greater in the species belonging to the genus Salmo than in that of the genus Salvelinus . The increase in the haematocrit values seems to have resulted from swelling of the erythrocytes and their decrease is similarly attributable to shrinking of the cells. The reasons for this swelling, which may be complicated, and its apparent significance for haematocrit determinations are discussed.     

Intestinal helminths in rainbow trout, Salmo gairdneri (Richardson): Absence of effect on nutrient absorption and fish growth

Rainbow trout. Salmo gairdneri (Richardson), from a fish farm, were found to be infected with two cestodes, Eubothrium crassum and Proteocephalus sp. Observations are described which suggest that these parasites have no effect on fish growth or on the intestinal absorption of L-leucine and D-glucose.     

Adult Triploids in a Rainbow Trout Family

Six triploid individuals were found in a full-sib family of 11 adult rainbow trout (Salmo gairdneri) from a domesticated hatchery stock. The triploid individuals were normal in size and external appearance, had underdeveloped gonads, and showed no evidence of 3n/2n chimerism or mosaicism. XXY triploids were males, suggesting that the Y chromosome is male determining in trout. Because they may avoid production losses associated with sexual maturation in normal fish, triploid trout and salmon could potentially be useful in fish culture.     

Use of lesion filtrates for the transmission of UDN (ulcerative dermal necrosis) in salmonids

Spawning Atlantic salmon, Salmo salar (L.), brown trout, Salmo trutta (L.) and rainbow trout, Salmo gairdneri (Richardson) were held in tanks for four weeks and then injected with filtrates from lesions of salmon naturally infected with UDN. In some cases prednisolone acetate was included in the inoculum. Water temperatures were recorded daily and the time of appearance of the lesions was noted. Passage experiments using extracts of these lesions were carried out.