Proliferative kidney disease: first reported outbreak in North America

Abstract. Proliferative kidney disease (PKD) was diagnosed in rainbow trout, Salmo gairdneri Richardson, at the Hagerman, Idaho, State Fish Hatchery. Wild fish from Riley Creek, one of the hatchery water supplies, were infected with PKD organisms. Clinical signs, gross pathology and histopathology of affected fish were typical of those reported for PKD in salmonids from European hatcheries. In addition to lesions normally seen, we observed a severe vasculitis and occlusion of renal and hepatic vessels. Ultrastructure of the PKD organism and possible inclusion body production sites are described. The disease outbreak was complicated by the presence of other infectious diseases. This is the first recorded outbreak of PKD in North America.     

A study of the sequential clinical and pathological changes during proliferative kidney disease in rainbow trout, Salmo gairdneri Richardson

Abstract. A histological study of 0+ rainbow trout, exposed to proliferative kidney disease (PKD) on an infected farm, showed evidence of infection from the beginning of June, gross pathological changes from the beginning of July, and clinical signs from mid-July. Evidence of healing was present in some samples from the second week of August. Kidney samples were divided into five grades (0–4) representing increasing severity of pathology. A sixth grade (H) was applied to kidneys showing signs of healing. The most frequent clinical signs were abdominal distension and exophthalmos. Melanosis, corneal cloudiness and ecchymoses were observed occasionally. Apart from renal enlargement, other internal gross pathological changes included ascites, gill and liver pallor, oedema, petechiation, and splenic enlargement. The majority of clinical signs and internal changes were only seen in fish with advanced renal swelling (grades 3 and 4). Histopathology was most marked in the kidney and was of three types: haemopoietic hyperplasia, vascular pathology and diffuse inflammatory change. Vascular changes, including occlusion of blood vessels in several organs and apparent haemoglobin crystallization, and destruction of renal excretory structures within areas of chronic diffuse inflammation were marked features of the histopathology in fish with clinical signs. The pathogenesis of PKD is discussed with reference to the histopathological changes.     

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.     

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.     

Effects of the probiotic, Lactobacillus acidophilus, on the growth performance, haematology parameters and immunoglobulin concentration in African Catfish (Clarias gariepinus, Burchell 1822) fingerling

This experiment was carried out to evaluate the effects of the probiotic, Lactobacillus acidophilus , on the growth performance, haematology parameters and immunoglobulin concentration in African catfish Clarias gariepinus fingerling. Two experimental diets were formulated to contain 35 g kg⁻¹ crude protein and 10 g kg⁻¹ lipids accordingly and fed three times daily for 12 weeks to 25 C. gariepinus fingerlings per fibreglass tank in 12 replicates each. The control diet was prepared with no probiotic supplementation whereas the second diet was prepared supplemented with a probiotic, L. acidophilus , containing about 3.01 × 10⁷ colonies/g of diet. The results show that growth performance [specific growth rate (SGR) and relative growth rate (RGR)], nutrient utilization [protein efficiency ratio (PER) and feed conversion ratio (FCR)] and survival were significantly ( P <0.05) higher in fish maintained on the probiotic-supplemented diet compared with those on the control diet. Haematology parameters (packed cell volume, haemoglobin, erythrocyte sedimentation rate, red blood cell and white blood cell, total serum protein, Ca²⁺, Mg²⁺, Cl⁻, glucose and cholesterol) and total immunoglobulin concentrations were also significantly better in fish fed the probiotic-supplemented diet than in the control. Although the water quality parameters monitored were better in the fish fed the probiotic-supplemented diet than in the control, the parameters were not significantly different ( P >0.05). From the results of this experiment, we conclude that L. acidophilus can be used as a probiotic agent in African catfish culture, to enhance fish health, survival and better feed efficiency and growth performance.     

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.     

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.     

The occurrence of proliferative kidney disease (PKD) in cultured and wild fish: further investigations

Abstract. In an investigation of the occurrence of proliferative kidney disease (PKD) in freshwater fish other than rainbow trout, 18 species of wild fish and seven species of fish raised in cultivation wore sampled from waters where the disease occurred annually in rainbow trout, Oncorhynchus mykiss (Richardson). Results revealed that certain wild stocks of brown trout. Salmo trutta L., grayling, Thymallus thymallus L., and pike, Esox lucius L., were infected with PKD, as were cultivated Atlantic salmon, Salmo salar L., parr, brown trout and char, Salvelinus alpinus (L.). Microscopical examination revealed the presence of the PKX cell in these species and also intraluminal protozoa possibly related to the PKX cell, which were not found in the rainbow trout. Other species of freshwater fish had myxosporidan infections but, unlike PKD infection, there was little host/parasite tissue response. The PKX cell as a myxosporidan stage is discussed and the presence of the disease in wild fish is reported.     

Development of proliferative kidney disease in rainbow trout, Oncorhynchus mykiss (Walbaum), following short-term exposure to Tetracapsula bryosalmonae infected bryozoans

The initial site of infection in the fish host for Tetracapsula bryosalmonae , causative agent of proliferative kidney disease (PKD) is poorly understood. Following the recent recognition that freshwater bryozoans harbour the infective stages to salmonid fish, experimental transmission studies were undertaken to investigate (1) the route of entry of the parasite into the fish host and (2) the minimum exposure time required to induce clinical signs of PKD. In-situ hybridization (ISH) studies were carried out on naïve rainbow trout exposed to the naturally infected bryozoan Fredericella sultana for up to 90 min. The sporoplasm of T. bryosalmonae was detected entering the fish via mucous cells in the skin epithelium within the first minute of exposure. In addition, T. bryosalmonae cells were infrequently detected in the skeletal musculature of exposed experimental fish up to 72 h post-exposure. The route of migration through the fish to the kidney and spleen was not determined. All fish exposed to infected, disrupted bryozoans for 10, 30 and 90 min and maintained for up to 8 weeks developed clinical PKD.     

Haematological and blood biochemical characteristics of <Emphasis Type="Italic">Glyptosternum maculatum</Emphasis> (Siluriformes: Sisoridae) in Xizang (Tibet)

This study investigated the haematological and blood biochemical characteristics of Glyptosternum maculatum. The haematological and biochemical parameters were measured in 30 adult fish collected from Nyingchi Reach of Yarlung Zangbo River in Tibet. The red blood cell count (RBC), haemoglobin concentration (Hb), haematocrit (Hct), erythrocyte osmotic fragility (maxEof and minEof), the erythrocyte sedimentation rate, mean cell volume (MCV), mean cellular haemoglobin content (MCH), and mean cell haemoglobin concentration (MCHC) were determined. Compared with other Siluriformes fishes, G. maculatum showed similar mean values for Hct, Hb, MCH, and MCHC and had slightly lower RBC and higher MCV. The biochemical parameters were assayed including alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase, total protein, albumin, globulin, albumin/globulin ratio, total bilirubin, direct bilirubin, urea, creatinine, glucose, total cholesterol, and triglyceride. The result showed that the value of AST in G. maculatum was obviously higher than that in Rhamdia quelen as well as in Silurus merdionalis.     

Post-haemorrhagic anaemia in sea bass,Dicentrarchus labrax (L.), caused by blood feeding of Ceratothoa oestroides (Isopoda: Cymothoidae)

The effects of the fish parasitic isopod, Ceratothoa oestroides (Risso), on haematological parameters of its cage-cultured sea bass host, Dicentrarchus labrax (L.), were studied. Analyses of blood parameters (cell counts, haemoglobin content and haematocrit) were carried out on parasitized and unparasitized sea bass from a fish farm in Turkey. Parasitized fish had significantly lowered erythrocyte counts, haematocrit and haemoglobin values and significantly increased leucocyte counts. Blood feeding by C. oestroides thus produces a post-haemorrhagic anaemia and the fish appear to mount an immune response to the presence of parasites.     

What determines the size of teleost erythrocytes? Correlations with oxygen transport and nuclear volume

An inverse relationship exists between erythrocyte size and the aerobic swimming ability of teleosts. A functional basis for this relationship is found in larger surface area to volume ratios and shorter diffusion distances allowing more rapid oxygen transfer as erythrocyte volume decreases. However, this apparently simple functional relationship is confounded by a more general direct relationship between cell size and nuclear volume. Nuclear DNA content, which is directly proportional to nuclear volume, is known to vary at least 11 fold among teleost species. Therefore a question arises as to the relative contributions of nuclear volume and the demands of the blood oxygen delivery system in determining erythrocyte volume.These relationships were investigated by regressing erythrocyte volume against nuclear volume and whole blood haemoglobin concentration (as an index of aerobic scope) for 52 species of teleost fish from 25 families. This analysis shows that erythrocyte volume is more highly correlated with oxygen delivery than with nuclear volume. However, much of the variation in erythrocyte volume is unexplained.     

大黄鱼(Pseudosciaena crocea)内脏白点病的组织病理和超微病理分析

利用组织切片及超薄切片电镜技术对患内脏白点病大黄鱼(Pseudosciaena crocea)的肝脏、脾脏和肾脏3种组织进行病理学分析,探讨该病的致病机理.结果显示,大黄鱼的临床症状为体表无明显病症,脾、肾、肝等内脏有大量白色结节;组织病理显示,肝、肾和脾是感染损伤的主要靶器官,出现组织变性坏死,空泡化严重,炎性细胞浸润;病变组织均出现病理性结节.超微病理显示,病鱼肝、肾、脾细胞超微结构受损严重,尤以线粒体和细胞核损伤明显.线粒体肿胀,嵴断裂消失,空泡化;细胞核核膜破裂,染色质浓缩边集;肾脏和脾脏均发现大量菌聚集成团的病原.研究表明,大黄鱼内脏白点病的组织细胞病理变化特征显示了病原菌的入侵和危害,造成鱼体生理代谢紊乱,免疫力和抵抗力下降,鱼体终因无法维持正常的生命活动而导致死亡.     

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.     

A study of gross,histological and blood biochemical changes in rainbow trout,Oncorhynchus mykiss (Walbaum), with rainbow trout gastroenteritis (RTGE)

The mechanisms behind the pathogenesis of rainbow trout gastroenteritis (RTGE) are still unknown. This study examined the macroscopic and microscopic changes in trout with RTGE (RTGE+), as well as the blood chemistry. A total of 464 rainbow trout were sampled from 11 sites in the UK, comprising 152 RTGE+ fish and 330 random, apparently healthy fish. A case definition for RTGE was assessed by the analysis of its agreement with three laboratory tests: histopathology, packed cell volume and kidney bacteriology. Cluster analysis indicated the presence of three distinct presentations within the population of RTGE+ fish. Cluster A included gross signs associated with moribund RTGE+ fish, and clusters B and C identified gross signs consistent with concurrent diseases, notably furunculosis, enteric redmouth and proliferative kidney disease. The information gained was used to select RTGE+ fish without concurrent disease for the analysis of RTGE pathogenesis with blood biochemistry. This analysis revealed a severe osmotic imbalance and a reduced albumin/globulin ratio as indicatives of selective loss of albumin. These findings are compatible with a protein losing enteropathy.     

Effects of spirulina powder (Spirulina platensis) as a dietary additive on Oscar fish,Astronotus ocellatus: Assessing growth performance,body composition,digestive enzyme activity,immune‐biochemical parameters,blood indices and total pigmentation

In this study, the effect of spirulina meal (Spirulina platensis) as a feed additive on growth and physiological response of Oscar fish,Astronotus ocellatus, was assessed using four diets including control without spirulina supplementation, 13.75 g/kg spirulina (SP1), 27.50 g/kg spirulina (SP2) and 55 g/kg spirulina (SP3). One hundred and twenty fish (10.41 ± 0.25 g) were disturbed to twelve aquariums and fed to apparent satiation three times a day for 8 weeks. The result showed that the SP3 treatment had statistically better growth performance, feeding parameters, total protease activity, total protein and lipid contents than the control. Moreover, haemoglobin, haematocrit, mean erythrocyte haemoglobin and mean erythrocyte haemoglobin concentrations were improved considerably in the SP3 treatment compared with the control. The SP3 treatment showed higher total protein and albumin levels, but lower triglycerides, cholesterol levels and liver enzyme activities than the control. Also, serum complements (C3, C4), lysozyme activity and total carotenoid contents in fish skin significantly increased in the SP3 treatment compared with the control. The results of this study suggest that spirulina powder at the level of 55 g/kg of the diet can be used as a functional natural feed additive to improve the performance of Oscar fish.     

Glomerulonephritis and immunosuppression associated with dietary essential fatty acid deficiency in gilthead sea bream, Sparus aurata L., juveniles

An experiment was conducted to determine the effect of an essential fatty acid (EFA) deficient diet on growth, immune status and renal morphology of juvenile gilthead sea bream, using two diets: a control diet containing 2% (DW) of n-3 high unsaturated fatty acid (n-3 HUFA) and a diet formulated to be deficient in EFA for this species and containing 0.5% DW of n-3 HUFA (diet NFA). After 9 weeks of feeding the EFA-deficient diet fish showed a reduction in growth compared with the control group (107.48 +/- 9.14 and 123.14 +/-11.87 g final weight respectively). Fish fed the NFA diet showed a reduction in the erythrocyte volume together with increased erythrocyte fragility, haemoglobin content and red blood cell count in comparison with fish fed the control diet. EFA deficiency also reduced cellular immunity in terms of neutrophil activity and the number of circulating lymphocytes. The serum alternative complement pathway was markedly reduced in fish fed the EFA deficient diet. Fish fed the control diet showed glomeruli with a well-defined Bowman's space and normal renal tubes. However, up to 88% of fish fed the EFA-deficient diet showed alterations in renal morphology affecting at least 50% of the glomeruli, which showed extreme dilation of capillaries and occlusion of Bowsman's capsule. Mesangial proliferation and diffuse thickening of the capillary walls, as well as renal tube degeneration, were also observed.     

Jaundice disease in the farmed catfish hybrid, Clarias macrocephalus (Gunther) ×C. gariepinus (Burchell), in Thailand

Abstract. In recent years, jaundice disease in farmed hybrid catfish in Thailand has caused high levels of mortalities. Affected fish are lethargic and anorexic and show yellow pigmentation of the skin and gills. Internally, the spleen, kidney and gall bladder are enlarged, and the spleen, kidney, liver and body fat are a pale yellow colour. Most fish contain a yellow ascitic fluid in their abdomen. Histological examination of tissues from diseased fish revealed heavy deposits of haemosiderin and ceroid in the spleen, kidney and liver. Haematological measurements showed that haematocrit values, RBC count, haemoglobin concentration, MCH and MCHC were significantly decreased in affected fish while MCV, total bilirubin and direct reacting bilirubin were significantly increased. The results indicate that the fish are suffering from a haemolytic anaemia associated with lipoid liver degeneration. The disease is thought to be due to the feeding of rancid chicken viscera.     

团头鲂同源四倍体、倍间三倍体与二倍体红细胞的形态特征比较

通过比较同源四倍体、倍间三倍体与二倍体团头鲂（Megalobrama amblycephala）的红细胞、红细胞核的形态大小差异,探讨不同倍性团头鲂的红细胞形态遗传特征。结果表明：（1）团头鲂多倍体（同源4n和倍间3n）的红细胞数量显著低于二倍体（2n）团头鲂（P〈0．05）。其中,倍间三倍体团头鲂每毫升血液的红细胞数为二倍体的78．95％,而同源四倍体的红细胞数量下降幅度较大,仅为2n团头鲂的56．87％。（2）团头鲂同源四倍体、倍间三倍体的红细胞短径和长径、核短径和长径、红细胞表面积、红细胞体积和核体积均依倍性增加而显著增大（P〈0．05）;其中,又以核体积增大最为显著,同源四倍体、倍间三倍体的核体积分别为2n团头鲂的1．97倍和1．39倍。（3）团头鲂同源四倍体每毫升血液红细胞的总体积和总表面积,均明显低于二倍体和倍间三倍体团头鲂（P〈0．05）;倍间三倍体每毫升血液红细胞的总体积要多于二倍体团头鲂（P〈0．05）,但红细胞的总表面积在倍间三倍体与二倍体团头鲂间不存在显著差异（P〉0．05）。（4）在团头鲂同源四倍体中还观察到一定比例的异常红细胞现象。     

Effect of infestation with the isopod, Alitropus typus M. Edwards (Crustacea: Flabellifera: Aegidae) on the haematological parameters of the host fish, Channa striatus (Bloch)

Detailed accounts supported by experimental data are furnished regarding the haematological parameters of the host fish, Channa striatus infested by the isopod, Alitropus typus. Results demonstrate that the infested fish become anaemic through loss of blood as shown by a reduction in the total erythrocyte count, haemoglobin content and haematocrit. Considering the mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and volume index, the anaemia can be designated as macrocytic and hypochromic. The total and differential counts of white blood corpuscles of the infested fish indicate leucopenia coupled with lymphocytosis. The percentages of monocytes, neutrophils and macrophages, which were 0.65%, 0.39% and 1.04%, respectively, after 24 h of infestation, had increased to 2.31%, 4.91% and 2.02%, respectively, after 96 h of infestation and declined thereafter.