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.     

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.     

Bryozoans as hosts for Tetracapsula bryosalmonae, the PKX organism

Freshwater bryozoans have recently been identified as hosts of Tetracapsula bryosalmonae , the causative agent of proliferative kidney disease (PKD). To gain insight as to which bryozoans are consistently important hosts for T. bryosalmonae , we present the results of a broad scale survey of bryozoans in European and North American sites upstream from PKD outbreaks in feral, stocked and farmed salmonids. The bryozoan genera most commonly found in association with disease outbreaks are Fredericella and Plumatella , and the most common species are F. sultana (Blumenbach), F. indica Annandale, and P. emarginata Allman. The prevalence of mature sac stages of T. bryosalmonae was very low. Our survey data together with knowledge of bryozoan life cycles and the ecology of PKD allow inferences concerning unknown aspects of the life cycle of T. bryosalmonae .     

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.     

The persistence of infectivity of Tetracapsulabryosalmonae-infected water for rainbow trout, Oncorhynchus mykiss (Walbaum)

Proliferative kidney disease (PKD) is caused by the infection of susceptible salmonid fish with spores of the myxozoan Tetracapsula bryosalmonae , a parasite harboured and released by several species of bryozoans. Under natural conditions, PKD is a water-borne infection of fish, whose outcome and spatio-temporal dissemination depend on the viability of spores present in the water. In order to evaluate the duration of parasite infectivity, juvenile rainbow trout, Oncorhynchus mykiss , were exposed for 20 h to T. bryosalmonae -infected water at various times post-water collection or after different filtration procedures. When infected water was held in a temperature range of 14.5–17 °C for up to 14 days, PKD was transmitted to the fish only between 0 and 12 h post-water collection and its infectivity vanished between 12 and 24 h. Similarly, the infectivity of water passed through 25 μm but not through 1 μm mesh filters, and was lost in the material eluted from the 1 μm filtration membrane although the parasite's DNA was amplified from this material. The parasitic infectivity in water appears to be fragile and this may offer opportunities to decrease the impact of PKD in trout farms by the implementation of management procedures aimed at reducing the number of the bryozoan-holding surfaces located in the river, immediately upstream from these farms.     

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: 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.     

高碳水化合物水平日粮对异育银鲫生长、生理、免疫和肝脏超微结构的影响

为了探讨异育银鲫对日粮中高碳水化合物的利用,选用168尾异育银鲫,分成2组,一组为正常对照组,投喂35%碳水化合物(35%CHO)水平日粮;另一组为高糖试验组,投喂50%碳水化合物(50%CHO)水平日粮每组设3个重复。在控温的循环水系统中饲养10周后,测定鱼体生长指标、肌肉成分、肝糖元、血液生化指标和免疫指标。结果表明,高糖组(50%CHO)显著(P<0.05)降低了特定生长率、增重率,但是对饵料系数、肝体比、脏体比、鱼体肌肉成分等没有显著(P>0.05)影响;两组异育银鲫血浆总蛋白、甘油三酯、总胆固醇、血糖和肝糖原含量都没有显著差异(P>0.05);高糖组(50%CHO)肝脏丙二醛含量显著升高(P<0.05)、天门冬氨酸转氨酶显著降低(P<0.05),而肝脏超氧化物歧化酶、总抗氧化能力、丙氨酸转氨酶和碱性磷酸酶都没有显著变化(P>0.05)。试验还对两组异育银鲫肝细胞进行超微结构观察,发现高糖饲料引起了异育银鲫肝细胞中糖原和脂肪的沉积,并对细胞和细胞器的结构产生一定损伤。从本试验结果来看,异育银鲫可以耐受50%饲料糖含量,但是长期摄食50%碳水化合物日粮对其生长不利,并会引起一定程度肝脏功能和结构的损伤。     

驼背鲈肝脏结构的光镜和透射电镜观察

采用HE、Mallory氏三色法、Heidenhain氏Azan染色,PAS反应,组织块银浸染方法,对组织切片染色处理,应用光学显微镜和透射电镜对驼背鲈肝脏的显微和超微结构进行了观察,结果表明驼背鲈肝组织中肝细胞界限不明显,不存在双核现象,有暗细胞和亮细胞之分,胞质中细胞器丰富,有大量的线粒体、内质网和溶酶体,高尔基体很难观察到,仅在胆小管和狄氏间隙的附近发现。此外,在肝血窦的附近观察到成纤维细胞、枯否氏细胞和贮脂细胞。肝组织中胆管数量众多包括肝叶中央胆管、支胆管、小叶间胆管和小叶内胆管,并且在中央静脉附近偶见辐射状排列的肝细胞索。     

管角螺性畸变现象的组织解剖学研究及扫描电镜观察

为了探讨管角螺性畸变过程中,生殖系统在形态和组织结构方面的变化,以及性畸变可能对其种群带来的不良影响,实验采用组织解剖学和扫描电镜的方法,研究了广东湛江、广西北海、钦州和防城港及越南胡志明市5个区域管角螺的未成熟和成熟正常雄、雌个体以及性畸变个体的生殖系统结构。结果显示,管角螺性畸变个体除具有正常的雌性生殖器官外,还有输精管和阴茎等雄性生殖器官,并且性畸变个体阴茎的位置、形状和输精管的组织结构与正常雄性个体的相似,管角螺性畸变现象仅见于成熟个体,高程度性畸变雌体的性腺发育受阻。性畸变率(IOI)结果显示,8批样中管角螺性畸变严重程度依次是北海1北海2湛江钦州1防城港近海越南胡志明市、钦州2和防城港白龙珍珠湾,其中北海的性畸变情况最为严重,性畸变率平均达到90%;北海种群的输精管发育指数(VDSI)也高于其他地区群体;并且北部湾管角螺的雌雄性比几乎都要小于1,总体呈现减少的趋势。研究表明,有机锡污染可能已对北部湾管角螺种群的延续造成威胁。     

长江口纹缟虾虎鱼精子、卵子及受精过程扫描电镜观察

采用电镜对长江口纹缟虾虎鱼成熟精子、卵子及受精早期精子入卵过程进行了观察。结果显示:纹缟虾虎鱼成熟精子由头部和尾部两部分组成。头部呈圆形或近圆形,无顶体,细胞核长径为1.15±0.28μm,短径为0.97±0.22μm,尾部鞭毛长为8.17±1.12μm。核膜外具有双层质膜,质膜表面不平整,在核膜和双层质膜中间有较大空隙存在。中心粒复合体位于植入窝内。袖套两侧分布有2～3个较大的线粒体。鞭毛为典型的"9+2"结构,有侧鳍。成熟卵呈圆形,卵膜表面多沟和嵴,具单一受精孔,孔洞区外径约5μm,孔内壁呈螺旋嵴。在卵的植物极有一盘状突起,突起的中间为圆形,周边有呈网状结构的粘丝与卵壳膜连接。纹缟虾虎鱼精子入卵速度较快,受精过程较短,受精后10～18 s完成精子入卵过程。     

Effects of azithromycin on tilapia (Oreochromis niloticus): health status evaluation using biochemical,physiological and morphological biomarkers

Bacterial diseases cause tilapia's high‐mortality outbreak. This study investigated the toxicity of azithromycin (AZT), a macrolide antibiotic that has been considered a possible therapeutic drug for tilapia aquacultural use. The 48‐h acute toxicity (50% lethal concentration, LC50; 48 h) of AZT was determined for Oreochromis niloticus. Thereafter, fish were exposed to 0, 1, 50 and 100 mg L^?1 AZT during 14 days (chronic exposure) and measured the haematological variables, the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione‐S‐transferase (GST) and the concentration of glutathione (GSH), protein carbonyl and lipid peroxidation in the liver; histopathology was analysed the liver, gills and kidneys. The LC50; 48 h was >100 mg L^?1. No fish died during chronic exposure. Haematocrit and haemoglobin concentration increased in fish exposed to 50 and 100 mg L^?1, and the total number of leucocyte and thrombocyte increased after exposure to 100 mg L^?1 AZT, suggesting a stimulation of defence cell production. In the liver, the antioxidant enzyme activities did not change, but GST activity and the GSH level increased in fish exposed to 100 mg L^?1 AZT. Oxidative stress did not occur. Histopathological index (HI_L) indicates moderate liver damage; minor histological changes in the gill and no change in the kidneys. AZT was considered non‐toxic for O. niloticus after acute exposure and, although it causes moderated histopathology in the liver after chronic exposure, this antibiotic may be an alternative against bacterial infections, depending on its efficacy to control bacterial disease in fish.     

The effect of feeding the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on cell-mediated immunity and protection against Yersinia ruckeri in pikeperch (Sander lucioperca)

The present study examined the influence of leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB), a natural product HMB, on nonspecific cell‐mediated defence mechanisms and protection against enteric redmouth disease (ERM) in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelletted ration at 50 mg kg^?1 of the feed for 8 weeks. The phagocytic ability and potential killing activity of blood and pronephric phagocytes were examined in HMB‐ and control‐fed fish before and after 8 weeks of feeding HMB. Simultaneously, the proliferative response of blood and pronephric lymphocytes stimulated by mitogens concanavaline A and lipopolisaccharide were examined in experimental and control groups. Following 8 weeks of HMB feeding, a challenge test was performed by injecting the fish with live pathogenic bacteria Yersinia ruckeri. β‐hydroxy‐β‐methylbutyrate applied in the diet for 8 weeks prompted a statistically significant (P<0.05) increase in phagocytic ability and potential killing activity of the blood and pronephric phagocytes and the proliferative response of blood and pronephric lymphocytes. The changes in these mechanisms correlated with protection against Y. ruckeri, the ERM pathogen. The results showed that feeding HMB increased the nonspecific cell‐mediated defence mechanisms and protection against ERM by reducing cumulative mortality (30%) following the challenge with pathogenic bacteria. Future studies will include determination of optimal doses and protocols of oral application of HMB to maximize the immunomodulatory effects and protection against viral diseases in intensive pikeperch culture.     

Experimental evidence on prevention of infection by the ectoparasitic protozoans Ichthyobodo salmonis and Trichodina truttae in juvenile chum salmon using ultraviolet disinfection of rearing water

In northern Japan, juvenile chum salmon Oncorhynchus keta (Walbaum) are released from hatcheries to enhance the fishery resource. Infections with ectoparasitic protozoans, particularly the flagellate Ichthyobodo salmonis and the ciliate Trichodina truttae, occasionally cause severe mortality among hatchery‐reared juveniles. This study examined the susceptibility of the two parasites to wide‐ranging UV irradiation (experiment 1) and then investigated whether UV disinfection of the rearing water using a commercial device was useful for preventing infections among juveniles in a small‐scale rearing system over a 28‐day period (experiment 2). In experiment 1, parasite mortality reached 100% with UV irradiation doses of ≥9.60 × 10⁵ μW s/cm² for I. salmonis and ≥8.40 × 10⁵ μW s/cm² for T. truttae. In experiment 2, disinfection of the rearing water at a UV irradiation dose of 2.2 × 10⁶ μW s/cm² succeeded in complete prevention of both parasites in the juvenile salmon. These results elucidate the minimum dose of UV irradiation for inactivation of I. salmonis and T. truttae, and demonstrate the usefulness of water disinfection using a commercial UV irradiation device to prevent infections by these parasites in hatchery‐reared juvenile chum salmon.     

Replacement of fish meal in juvenile channel catfish,Ictalurus punctatus,diets using a yeast‐derived protein source: the effects on weight gain,food conversion ratio,body composition and survival of catfish challenged with Edwardsiella ictaluri

We examined the effects of a yeast‐derived protein source (NuPro^®) as a replacement for menhaden fish meal on weight gain, specific growth rate (SGR), food conversion ratio (FCR), whole‐body composition and disease resistance in juvenile channel catfish (9.9 ± 0.2 g fish^?1). NuPro^® replaced fish meal at six levels (0, 25, 50, 75, 100 and 125 g kg^?1 diet). Catfish were sampled for whole‐body composition and then challenged with the bacterium Edwardsiella ictaluri. Growth performance was negatively affected (P < 0.01) when NuPro^® was added at 125 g kg^?1 diet. The amount of whole‐body fat decreased (P < 0.05) when NuPro^® was added at 75 g kg^?1 or more of the diet. Regardless of the amount of NuPro^® added, survival after challenge with E. ictaluri was similar among treatments. Results indicate that up to 100 g kg^?1 of NuPro^® can be added without negatively affecting growth performance. The yeast‐derived protein source used in this study is a sustainable protein alternative that could be used as a partial replacement for fish meal in juvenile channel catfish diets.