Influence of water quality on the outbreak of proliferative kidney disease – field studies and exposure experiments

This paper presents data from field studies and exposure experiments and the possible association of limno- and physicochemical parameters with outbreaks of proliferative kidney disease (PKD) in rainbow and brown trout. The investigations were carried out at Singold Brook in southern Germany. Exposure experiments and sampling of wild fish were performed in Singold at least twice a year in 1992, 1994, 1996 and 2000. Both wild fish collected from the specific sites and experimentally exposed rainbow trout were investigated histologically for the occurrence of PKD. At the time of sampling, various water parameters at the respective river sites were measured. The results indicate strongly the existence of a correlation between organic pollution of water, the presence of Bryozoa and the outbreak of PKD.     

Distribution and prevalence of T. bryosalmonae in Austria: A first survey of trout from rivers with a shrinking population

The first evidence of proliferative kidney disease (PKD) in an Austrian river (the River Kamp) was documented in 2016, and no information on the PKD infection status of trout in other rivers was available. Since then, brown trout (Salmo trutta fario) and rainbow trout (Oncorhynchus mykiss) have been collected from rivers in Upper and Lower Austria for different diagnostic purposes. In this study, we summarize the recent findings of a first survey concerning the distribution of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), from these samples. Between September 2015 and October 2017, a total of 280 brown trout and 39 rainbow trout were collected from 21 rivers in the provinces of Upper and Lower Austria. T. bryosalmonae was detected by PCR of kidney tissue in 17 of 21 sampled rivers and in 138 of 280 brown trout as well as in 11 of 39 rainbow trout. Pathological signs of PKD (e.g., hypertrophy of the kidney) were observed in 33 analysed brown trout and six rainbow trout samples. No correlations between fish infected by T. bryosalmonae and the parameters size and age class, condition factor, geological origin of the streams and distribution within the river course were found, while positively tested fish are significantly increased at sampling sites exceeding water temperatures of 15°C for median periods of 115 days. The prevalence within the affected streams or stream sections is highly variable, and in single rivers, infection rates of up to 90% are confirmed.     

Proliferative kidney disease in Switzerland: current state of knowledge

This study presents an overview of the distribution of proliferative kidney disease (PKD) in wild and farmed fish in Switzerland based on two sources: (1) the results of routine diagnostic work at the National Fish Disease Laboratory, and (2) the results of a country‐wide survey for PKD. The first case of PKD in Switzerland was diagnosed in 1979 and since 1981, a few cases have been found every year. Affected species were rainbow trout, brown trout and grayling from rivers and fish farms. Most of the diseased fish were found in the lower altitude regions of the country (Swiss midlands). A seasonal distribution of the cases was evident: PKD‐positive fish occurred only from June to November withprominent peaks in August and September. Among infected fish all length classes were present, but the highest numbers of affected fish were found in the length classes <10 cm and 16–20 cm. The evaluation of the samples from a survey of the occurrence of PKD in 2000/2001 revealed a much broader geographical distribution of the infection than expected from routine diagnostic observations. Fish from 56 of 139 rivers analysed by histological examination were positive for parasites. Fish from 131 of these sites were examined macroscopically. Of these, trout from 45 sites showed gross alterations indicative for PKD. Most of the positive fish were found in the Swiss midlands north of the Alps. Fish from six of 65 fish farms investigated were found to be PKD‐positive by histopathological analysis, and in four of these cases fish could be identified as infected on the basis of gross examination.     

A survey on Tetracapsuloides bryosalmonae infections in Slovene fresh waters

Slovenia has no history of health problems related to proliferative kidney disease (PKD) either in farmed or in wild fish. However, due to the past molecular evidence for the presence of Tetracapsuloides bryosalmonae DNA in tissues of some fish from open waters, a survey was conducted on wild salmonids that were primarily sampled for other purposes. In winter 2010–2011, specimens from a total of 244 rainbow trout, Oncorhynchus mykiss (Walbaum), and brown trout, Salmo trutta L., from 30 bodies of fresh water were examined for T. bryosalmonae using a PCR method. The sampled fish showed no clinical signs or gross pathological lesions characteristic of PKD. Nineteen (7.8%) fish from seven (23.3%) fresh waters were positive for T. bryosalmonae. The identity of PCR amplicons was confirmed by sequencing. With one exception, all the positive fish were found in waters from the regions where the average yearly temperatures and the environmental pollution are higher. This preliminary countrywide survey provided the first insight into the situation regarding T. bryosalmonae infection of wild salmonids in Slovenia.     

Proliferative kidney disease in brown trout, Salmo trutta L., and Atlantic salmon, Salmo salar L., parr: histopathological and epidemiological observations

Abstract Proliferative kidney disease (PKD) has been described in cultivated rainbow trout but major outbreaks have not been recognized in other fish species. This paper describes light and electron microscope studies and presents epidemiological data of a disease condition in first summer Salmo salar parr and S. trutta , which is associated with an organism apparently identical to the aetiological agent of PKD. The pathology was found to be similar to that described for PKD in rainbow trout. The epidemiology in the brown trout suggests that exposure after May does not result in infection. The data from the outbreaks in two salmon hatcheries appear complex and suggest different susceptibilities to PKD among salmon originating from different sources.     

Are brown trout Salmo trutta fario and rainbow trout Oncorhynchus mykiss two of a kind? A comparative study of salmonids to temperature‐influenced Tetracapsuloides bryosalmonae infection

Proliferative kidney disease (PKD) of salmonids caused by Tetracapsuloides bryosalmonae causes high mortalities of wild brown trout (Salmo trutta fario) and farmed rainbow trout (Oncorhynchus mykiss) at elevated water temperatures. Here the aim was to compare the temperature‐dependent modulation of T. bryosalmonae in the two salmonid host species, which display different temperature optima. We used a novel experimental set‐up in which we exposed brown trout and rainbow trout to an identical quantified low concentration of T. bryosalmonae for a short time period (1 hr). We followed the development of the parasite in the fish hosts for 70 days. PKD prevalence and parasite kinetics were assessed using qPCR. Exposures were performed at temperatures (12°C and 15°C) that reflect an environmental scenario that may occur in the natural habitat of salmonids. T. bryosalmonae infection was confirmed earliest in brown trout kept at 15°C (day 7 post‐exposure) while, in all other groups, T. bryosalmonae was not confirmed until day 15 post‐exposure. Moreover, significantly greater infection prevalence and a faster increase of parasite intensity were observed in brown trout kept at 15°C than in all other groups. These results indicate that PKD is differentially modulated by water temperature in related host species.     

Immunohistochemical and PCR studies of wild fish for Tetracapsula bryosalmonae (PKX), the causative organism of proliferative kidney disease

Tetracapsula bryosalmonae, previously referred to as PKX, causes proliferative kidney disease (PKD) in salmonids and is an economically important myxozoan pathogen in salmonid culture. A variety of molecular and immunological tools have been developed to detect the parasite. To determine the specificity of four monoclonal antibodies (MAbs) raised against T. bryosalmonae, archive material of fish infected with various myxosporean species was obtained and immunostained. Wild fish were also collected from enzootic waters and examined for T. bryosalmonae infection using immunohistochemistry and the polymerase chain reaction (PCR). Three of the MAb probes appear to be specific for T. bryosalmonae while only two of the five sets of primers tested appeared to specifically amplify T. bryosalmonae DNA. The results of the immunostaining and the PCR demonstrate that T. bryosalmonae occurs in the tubules of grayling Thymallus thymallus L., brown trout, Salmo trutta L. and Atlantic salmon, Salmo salar L. outside of the PKD season (June‐September) in the UK. This confirms the results of previous studies that these species are the preferred fish hosts for the parasite in the UK.     

Competitive interactions between native and exotic salmonids: a combined field and laboratory demonstration

Abstract –  We studied the impact of two exotic salmonid species (brook trout, Salvelinus fontinalis and rainbow trout, Oncorhynchus mykiss ) on native brown trout ( Salmo trutta fario ) habitat, growth and survival. Habitat selection and vertical distribution between young-of-the-year of the three species were examined in a stream aquarium under different sympatric and allopatric combinations. In addition, similar species combinations were introduced in a Pyrenean mountain stream (southwest France) in order to extend laboratory results to growth and apparent survival. Both laboratory and field results indicated that rainbow trout significantly affected native brown trout habitat selection and apparent survival. On the contrary, brown trout habitat, growth and apparent survival were hardly affected by brook trout. These results support the idea that rainbow trout negatively influence native brown trout, and that competition could influence the outcome of fish biological invasions in freshwater ecosystems.     

Lectin histochemical studies on Sphaerospora sp. (Myxosporea) from Italian brown trout, Salmo trutta L.

A Sphaerospora sp. (Myxosporea) infection (presumably S. truttae ) was identified on a trout farm in northeastern Italy. Parasites were detected in kidneys from infected brown trout, Salmo trutta L., over a 2-year period. Extrasporogonic, sporogonic stages and mature spores were simultaneously detected in the same fish. Traditional diagnostic methods for Sphaerospora spp. rely on the detection of the myxosporean developmental stages in Giemsa-stained kidney smears or haematoxylin-eosin stained tissue sections. A histochemical method was employed where 10 biotinylated lectins (Con-A, DBA, SBA, GS-I, PHA-P, LEA, PWM, RCA₁, WGA and UEA-I) and the avidin-biotin-peroxidase complex (ABC) were used on Sphaerospora -infected brown trout renal tissues and kidney imprints. Five monoclonal antibodies against PKX (Mab12, MabA3, MabC5, MabD4 and MabB4) were also tested. A lectin glycoconjugate binding pattern for Sphaerospora spp. is presented. This staining method shows that SBA lectin ( Glycine max agglutinin) is a useful tool for the detection of the Sphaerospora spp. Only MabB4 bound some of the most mature sporogonic stages. In contrast Mabs12, A3, C5 and D4, and GS-I lectin ( Griffonia simplicifolia agglutinin) did not stain any of the Sphaerospora spp. stages, but did bind very specifically to the sporogonic and extrasporogonic stages of PKX, the causative agent of proliferative kidney disease (PKD).     

Proliferative kidney disease: cellular aspects of the rainbow trout, Oncorhynchus mykiss (Walbaum), response to parasitic infection

Proliferative kidney disease (PKD) is an immunopathological condition of salmonid fish, caused by the hyperplastic response of their principal lymphoid tissues to infection with the spores of Tetracapsula bryosalmonae , a myxozoan parasite formerly designated proliferative kidney organism – unknown (PKX). In order to investigate the nature of cells involved in this host response and possible alterations of their functions during parasitic infection the course of PKD was studied by flow cytometry (FCM) techniques, using blood, pronephros and spleen leucocyte populations from rainbow trout infected by intraperitoneal (i.p.) injection with parasitic cells from infected donor fish. The parameters of the cellullar response studied were: cytogram of cell population, lymphoproliferation, phagocytosis, oxidative burst, and non-specific cytotoxicity. The modifications of cell population distribution and function in the PKD-infected fish mainly affected the pronephros cell populations and were coincident with the clinical phase of disease. During this phase, the lymphocytes constituted the major leucocyte cell population and underwent proliferation and were thus responsible for the renal tissue hyperplasia. Meanwhile, phagocytosis and oxidative burst were depressed. These data are in agreement with the patho-epidemiological background of PKD where the enhancement of the fish sensitivity to bacterial infections reflects the impairment of certain cellular defence mechanisms of innate immunity.     

Patterns of Tetracapsuloides bryosalmonae infection of three salmonid species in large,deep Norwegian lakes

Proliferative kidney disease (PKD), caused by the myxozoan endoparasite Tetracapsuloides bryosalmonae, is of serious ecological and economical concern to wild and farmed salmonids. Wild salmonid populations have declined due to PKD, primarily in rivers, in Europe and North America. Deep lakes are also important habitats for salmonids, and this work aimed to investigate parasite presence in five deep Norwegian lakes. Kidney samples from three salmonid species from deep lakes were collected and tested using real-time PCR to detect PKD parasite presence. We present the first detection of T. bryosalmonae in European whitefish in Norway for the first time, as well as the first published documentation of the parasite in kidneys of Arctic charr, brown trout and whitefish in four lakes. The observed prevalence of the parasite was higher in populations of brown trout than of Arctic charr and whitefish. The parasite was detected in farmed, but not in wild, charr in one lake. This suggests a possible link with a depth of fish habitat and fewer T. bryosalmonae-infected and PKD-affected fish. Towards a warmer climate, cold hypolimnion in deep lakes may act as a refuge for wild salmonids, while cold deep water may be used to control PKD in farmed salmonids.     

Field study indicating susceptibility differences between salmonid species and their lineages to proliferative kidney disease

Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) is the causative agent of proliferative kidney disease (PKD), which affects both wild and farmed salmonid fish. The objective of this study was to outline differences in susceptibility to PKD in different salmonid species, hybrids and breeding lineages. Susceptibility to T. bryosalmonae infection was established based on cumulative mortality, pathological findings and detection of T. bryosalmonae in the kidney using immunohistochemistry and molecular methods. Determination of pure and hybrid individuals of different species in the genus Salvelinus, and dissimilarity of rainbow trout lineages, was performed using traditional polymerase chain reaction (PCR) and microsatellite analyses. Rainbow trout displayed higher disease severity compared with brook trout and Alsatian charr. Moreover, the results indicated differences in infection susceptibility, not only among different salmonid species but also among different lineages of charr and rainbow trout. Our study indicated that some salmonid species and even different lineages of the same species are more suitable for farming under PKD pressure.     

Comparison of the performance of brown trout, Salmo trutta L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in a put-and-take fishery

Abstract. Stock and catch statistics for a small (5–5ha) lowland put-and-take trout fishery have been used to investigate the relative catchability of brown, Salmo trutta L., and rainbow, Oncorhynchus mykiss (Walbaum), trout. Brown trout were consistently less vulnerable to fly fishing than rainbows, and were particularly difficult to catch between mid-June and September. As a consequence, the turn-over of rainbow trout stock was more rapid than that of brown trout, but eventual recapture levels of both species were similar at around 90% of the fish stocked. In these circumstances, brown trout could be regarded as a long-term investment for fishery managers, with rainbows providing more'instant'sport.     

Susceptibility of selected freshwater fish species to a UK Lactococcus garvieae isolate

Gram-positive cocci recovered from diseased rainbow trout from a farm in England were characterized by different methods, including pulsed field gel electrophoresis, as virulent Lactococcus garvieae serogroup 2 (pulsotype A1). Groups of rainbow trout were kept at a range of temperatures and injected intraperitoneally (i.p.) with one of the UK isolates, L. garvieae 00021. The 18 °C and 16 °C groups showed 67% and 28% mortality, respectively, by day 27 post-injection. Fish kept at 14 °C or lower were less susceptible (≤3% mortality). Raising the temperature of all groups to 18 °C at day 27 post-injection did not result in recurrence of the disease, even though viable bacteria were recovered from all groups 42 days later. Grayling were highly susceptible, with 65% mortalities when challenged with 200 colony forming unit fish⁻¹ by i.p. injection and 37% mortalities when exposed to effluent water from tanks containing affected rainbow trout. Other fish species tested, Atlantic salmon, brown trout and seven cyprinid species, were less susceptible. Viable L. garvieae was isolated from the internal organs of all species tested at the end of the trials, suggesting that they may pose a threat as possible carriers to susceptible farmed and wild fish.     

A brown trout (Salmo trutta) population faces devastating consequences due to proliferative kidney disease and temperature increase: A case study from Austria

Proliferative kidney disease (PKD) is an emerging parasitic disease that affects wild and farmed salmonid fishes worldwide. Currently, it is widespread in Europe with records in many countries. This study assessed the status of PKD in the river Wulka, a small Austrian river, which was known to host a vital brown trout population despite having high water temperatures. Contrary to the initial hypothesis that the river was free from PKD, we observed an overall PKD prevalence of 92%. Noninfected fish were restricted to the uppermost river section. Twenty-two out of 87 PKD-positive fish of all age classes showed clinical signs, while five individuals exhibited signs of recovering from PKD infection. The severity of clinical signs was significantly dependent on the fish length and age, with smaller or younger individuals being more severely affected. Comparison of data from the earlier fish sampling events with those results from 2017 and 2018, together with the analyses of the pattern of water temperature since 2010, indicates that Wulka's brown trout population might become extinct in the near future.     

The bacterial flora of rainbow trout, Salmo gairdneri Richardson, and brown trout, Salmo trutta L., eggs and its relationship to developmental success

Abstract. The bacterial flora of rainbow, Salmo gairdneri Richardson, and brown trout, Salmo trutta L., eggs was studied during incubation. Few bacteria were considered to exist internally, although Pseudomonas species and Aeromonas hydrophila were isolated. During incubation, large numbers of bacteria approaching 500 colony forming units/mm² accumulated around egg surfaces. Both rainbow and brown trout eggs placed in a through-flow system were colonized mainly by Pseudomonas sp. and A. hydrophila , whilst rainbow trout eggs in a separate recycling system were dominated by a Cytophaga species. Glass beads were also incubated (as an inert surface) and found to support significantly lower numbers of bacteria. Egg mortalities were recorded daily and found to differ significantly between both fish species and water conditions. Statistical analysis indicated a potentially significant correlation between hatching success and numbers of surface bacteria.     

Occurrence and pathogenicity of Yersinia ruckeri at fish farms in northern and central Finland

Abstract. Salmonid fish at fish farms in northern and central Finland and perch, Perca fluviatilis L., roach, Rutilus rutilus (L.), and whitefish, Coregonus sp., from four lakes in central Finland were studied between 1985 and 1990 for the occurrence of Yersinia ruckeri. The bacteria were found in fish from both areas, but in most cases, only single diseased salmon, Salmo salar L., brown trout, S. trutta L., rainbow trout, Oncorhynchus mykiss (Walbaum), whitefish and perch were encountered and were always connected with stress conditions. One clinical outbreak occured in salmon fingerlings in northern Finland, and the fish were successfully treated with trimethoprim-sulpha. Monthly monitoring of lake fish revealed two symptomless carrier perch in two lakes. Outwith the main study a moribund perch with yersiniosis was found in a polluted lake, and for the first time in Finland, a rainbow trout was also found to have contracted yersiniosis in a small private pond. Sorbitol-positive and negative isolates have been found to occur in both moribund and carrier farmed fish, indicating that the sorbitol test is not essential when evaluating the pathogenicity of Y. ruckeri.     

A systemic protozoal disease of cultured salmonids

Abstract. Sporadic cases of a systemic protozoal infection with cells resembling Dermocystidium sp. are described from cultured rainbow trout, Salmo gairdneri Richardson, from France and brown trout, Salmo trutta L., and Atlantic salmon, Salmo salar L., from Ireland. Classical proliferative kidney disease was additionally found in the rainbow trout and PKX-like cells were also seen in the brown trout and salmon.     

Oral transmission as a route of infection for viral haemorrhagic septicaemia virus in rainbow trout, Oncorhynchus mykiss (Walbaum)

Surveys among wild marine fish have revealed occurrence of viral haemorrhagic septicaemia virus (VHSV) infections in a high number of diverse fish species. In marine aquaculture of rainbow trout, preying on invading wild fish might thus be a risk factor for introduction and adaptation of VHSV and subsequent disease outbreaks. Our objective was to determine whether an oral transmission route for VHSV in rainbow trout exists. Juvenile trout were infected through oral, waterborne and cohabitation transmission routes, using a recombinant virus strain harbouring Renilla luciferase as reporter gene. Viral replication in stomach and kidney tissue was detected through bioluminescence activity of luciferase and qRT-PCR. Replication was detected in both tissues, irrespective of transmission route. Replication patterns, however, differed among transmission routes. In trout infected through oral transmission, replication was detected in the stomach prior to kidney tissue. In trout infected through waterborne or cohabitation transmission, replication was detected in kidney prior to stomach or in both tissues simultaneously. We demonstrate the existence of an oral transmission route for VHSV in rainbow trout. This implies that preying on invading infected wild fish is a risk factor for introduction of VHSV into marine cultures of rainbow trout.     

The susceptibility of salmonid fish to an atypical strain of Aeromonas salmonicida that infects goldfish, Carassius auratus (L.), in Australia

Abstract. An enzootic, Australian, atypical strain of Aeromonas salmonicida isolated from diseased goldfish, Carassius auratus (L.), was inoculated into Atlantic salmon, Salmo salar L., brown trout, S. trutta L., rainbow trout, S. gairdneri Richardson, and brook trout, Salvelinus fontinalis (Mitchill), fingerlings by intraperitoneal injection (i.p.) and by bath challenge, the latter with and without prior abrasion of skin. The 10-day LD₅₀ (i.p.) was estimated to be 7·4 × 10^-3 colony forming units (cfu) for Atlantic salmon, 3·0 × 10^-2 cfu for brown trout, 3·7 × 10² cfu for brook trout and 6·4 × 10³ cfu for rainbow trout. Brown, rainbow and brook trout succumbed to bath challenges with between 10⁵–10⁶ cfu/ml, developing ulcers of the skin and septicaemia. The organism was trasmitted from inoculated fish to five of 195 within-tank control fish via water and established a carrier state in one of 14 Atlantic salmon. It was concluded that the organism poses a significant threat to the salmonid farming industry and wild salmonid fisheries in Australia.