Reno-, hepato- and splenomegaly of common carp fingerlings (Cyprinus carpio L.) diseased in swimbladder inflammation caused by Sphaerospora renicola Dyková et Lom, 1982

The weight of internal organs (swimbladder, kidney, liver, spleen) in relation to the body weight was studied in common carp fingerlings divided into three groups on the basis of swimbladder appearance and microscopic examination of the kidney. The fish had been collected from different Hungarian fish farms at the time when swimbladder inflammation (SBI) usually occurs (in July and August). The first group comprised fish with severe signs of SBI and massive renal sphaerosporosis, the second group consisted of fish with milder swimbladder changes and/or kidney infection by a low number of Sphaerospora renicola, while the third group was constituted by infection-free common carp fry. Statistical analysis of swimbladder, kidney, liver and spleen weight in relation to the body weight revealed that in the infected groups the internal organs were substantially enlarged. This suggests that in common carp fry with SBI the swimbladder changes are accompanied by reno-, hepato- and splenomegaly.     

New trends in parasitic infections of cultured freshwater fish

The 1980s are characterized by the appearance of new fish parasitoses, the clarification of the aetiology of diseases and new insights into the process of infection and the development of important parasites, especially myxosporidians. Two phaerosporoses have entered into the foreground: swimbladder inflammation (SBI) caused by Sphaerospora renicola in common carp and proliferative kidney disease (PKD) in salmonids. Both diseases must be reported in the German Democratic Republic (GDR). The connection between the "C-protozoa" in the blood and S. renicola in SBI was recently clarified by experiments by Molnar (1988). The results of our own investigations indicate that the "K-protozoa" phase of S. renicola parasitizes mainly the brain and sometimes the spleen, pancreas and liver before appearing in the swimbladder. Hedrick et al. (1988) in the USA and Odening et al. (1988b) in Europe first reported a Sphaerospora sp. in Salmo gairdneri in connection with PKD. The spectacular findings by Wolf and Markiw (1984) and El-Matbouli (1988) elucidating on the life cycle of Myxobolus cerebralis have not yet brought the revolutionary change expected for a general new estimation of myxosporidian transmission as a basis for ecotechnological control measures. Examples for the formation of new regionally specific parasite colonizations offer the far Eastern cyprinids introduced into Europe. In the GDR the silver carp is infected by Chloromyxum barbi (Myxosporidia) (Sedlazsek), the pathogenic Gyrodactylus sprostonae and two new Gyrodactylus species (Pectobathrii) (Lux). Eel parasites new to Europe are the obviously imported and highly pathogenic nematodes Anguillicola crassus and A. novaezelandiae (Paggi et al., 1982; Moravec and Taraschewski, 1988). In Central Europe the infestation of rainbow trout by the tapeworm Proteocephalus neglectus is increasing (Priemer, 1980; Hanzelova et al., 1988).     

Molecular phylogeny of the kidney-parasitic Sphaerospora renicola from common carp (Cyprinus carpio) and Sphaerospora sp. from goldfish (Carassius auratus auratus)

The genetic relatedness of two kidney-parasitic Sphaerospora species was studied. Although S. renicola, the causative agent of swimbladder inflammation of common carp fingerlings (Cyprinus carpio), and Sphaerospora sp. originating from goldfish (Carassius auratus auratus) were indistinguishable on the basis of spore morphology, they were found to be genetically different as their 18S rDNA sequences shared only 71.9% identical nucleotides. In the phylogenetic trees, Sphaerospora sp. from goldfish grouped with Myxidium truttae (AJ582061) within the clade of the coelozoic freshwater species. Sphaerospora renicola clustered with S. molnari (AF378345) within the group of myxosporeans histozoic in gills. The topology of the six Sphaerospora species on the phylogenetic trees implied that myxospore morphology does not correlate with the genetic relationships, and the genus seems to be polyphyletic.     

Protocol for assessing brain function in fish and the effectiveness of methods used to stun and kill them

A method for the evaluation of brain function in fish has been developed which is based on assessments of self-initiated behaviours, responses to stimulation, and reflexes. These assessments were validated in several freshwater and marine species and applied to evaluate the brain function of fish while they were anaesthetised and while they were being killed by a variety of methods. The results of these investigations were compared with published neurophysiological observations and it is concluded that some of the tests can be used to identify, with reasonable confidence, the state of awareness, and thus the capacity to experience suffering of several species of fish.     

Nutrition of ornamental fish.

Because thousands of species comprise the aquarium hobby, ornamental fish nutrition is an art and a science that must be approached systematically and holistically. Examinations of species-specific anatomy and natural history are useful starting points. Food fish research provides fundamental nutritional information, but food fish are not ideal models for all ornamentals. This article briefly addresses freshwater and marine fish issues, including considerations for pond fish and live foods.     

Global trade in ornamental fish from an Australian perspective: the case for revised import risk analysis and management strategies

Over 1 billion ornamental fish comprising more than 4000 freshwater and 1400 marine species are traded internationally each year, with 8-10 million imported into Australia alone. Compared to other commodities, the pathogens and disease translocation risks associated with this pattern of trade have been poorly documented. The aim of this study was to conduct an appraisal of the effectiveness of risk analysis and quarantine controls as they are applied according to the Sanitary and Phytosanitary (SPS) agreement in Australia. Ornamental fish originate from about 100 countries and hazards are mostly unknown; since 2000 there have been 16-fold fewer scientific publications on ornamental fish disease compared to farmed fish disease, and 470 fewer compared to disease in terrestrial species (cattle). The import quarantine policies of a range of countries were reviewed and classified as stringent or non-stringent based on the levels of pre-border and border controls. Australia has a stringent policy which includes pre-border health certification and a mandatory quarantine period at border of 1-3 weeks in registered quarantine premises supervised by government quarantine staff. Despite these measures there have been many disease incursions as well as establishment of significant exotic viral, bacterial, fungal, protozoal and metazoan pathogens from ornamental fish in farmed native Australian fish and free-living introduced species. Recent examples include Megalocytivirus and Aeromonas salmonicida atypical strain. In 2006, there were 22 species of alien ornamental fish with established breeding populations in waterways in Australia and freshwater plants and molluscs have also been introduced, proving a direct transmission pathway for establishment of pathogens in native fish species. Australia's stringent quarantine policies for imported ornamental fish are based on import risk analysis under the SPS agreement but have not provided an acceptable level of protection (ALOP) consistent with government objectives to prevent introduction of pests and diseases, promote development of future aquaculture industries or maintain biodiversity. It is concluded that the risk analysis process described by the Office International des Epizooties under the SPS agreement cannot be used in a meaningful way for current patterns of ornamental fish trade. Transboundary disease incursions will continue and exotic pathogens will become established in new regions as a result of the ornamental fish trade, and this will be an international phenomenon. Ornamental fish represent a special case in live animal trade where OIE guidelines for risk analysis need to be revised. Alternatively, for countries such as Australia with implied very high ALOP, the number of species traded and the number of sources permitted need to be dramatically reduced to facilitate hazard identification, risk assessment and import quarantine controls.     

Tropical fish medicine. Necropsy examination of fish

Necropsy examination of moribund or dead specimens is an essential step in diagnosing fish diseases. This article discusses basic anatomy and necropsy procedures. A large part of the article has been devoted to methods of procuring samples for laboratory analysis and histologic examination. To determine which findings are responsible for the mortalities, the entire case history and gross necropsy findings must be reviewed. Careful observations made during the necropsy examination provide valuable information immediately, as well as later on in the interpretation of laboratory data.     

Renal pathology in reptiles.

The class of Reptilia varies widely. Both the gross morphology and microscopic anatomy of the kidneys are specific for each species.In each species of reptile, the physiology of the renal system has adapted to the specific conditions of life, including, among other factors, the type of food, environmental temperature, and the availability of water. The pathology of the kidneys in reptiles has been poorly studied, but in recent years a number of investigators have specifically studied reptilian renal pathology.     

Multi-locus Sequence Analysis (MLSA) of Edwardsiella tarda isolates from fish

Edwardsiella tarda is an enteric fish pathogen that has caused significant economic losses in a range of fish species residing in diverse ecological conditions. Several molecular methods relying on DNA fingerprinting (RAPD, RFLP and ERIC-PCR) and the gyrB gene marker have been used to characterize E. tarda isolates. However, all had drawbacks in resolving power and reproducibility. The present study was aimed at developing a novel Multi-locus Sequence Analysis (MLSA) scheme for genetic characterization of E. tarda isolates originating from multiple sources. MLSA has been described as an effective molecular tool with superior discriminatory power and reproducibility for exploring intra-species genetic diversity of several bacterial species. Nucleotide sequence fragments of eight protein coding housekeeping genes (gyrB, mdh, adk, dnaK, phoR, metG, pyrG and aroE2) were obtained from 23 fish pathogenic E. tarda isolates of different geographical origins, one human isolate and 3 reference strains. The phylogenetic relationships between isolates in individual gene analyses were not consistent, although some common patterns were apparent. Phylogenetic analysis based on concatenated sequences of seven gene loci, however, buffered the conflicting phylogenetic signals and resolved isolates according to their geographical origin and/or fish host. The MLSA revealed two major genetically diverging clusters in E. tarda isolates examined, one cluster representing isolates from fish and the other representing (in the main) human isolates, with E. ictaluri cluster situated in between. The results suggest, therefore, that the fish pathogenic E. tarda isolates may have been previously misclassified and probably represent one or more as yet unrecognized taxa within the genus Edwardsiella. The MLSA described here was robust enough in discriminating E. tarda isolates not only with respect to their geographical origins but also within different hosts from the same geographical location, high-lighting its potential application in tracing the source of infection and understand the epidemiological relationships among isolates of environmental, fish, other domestic animals or human origins.     

Recent developments in fish vaccinology

During the last 10 to 20 years vaccination has become established as an important method for prevention of infectious diseases in farmed fish, mainly salmonid species. So far, most commercial vaccines have been inactivated vaccines administered by injection or immersion. Bacterial infections caused by Gram-negative bacteria such as Vibrio sp., Aeromonas sp., and Yersinia sp. have been effectively controlled by vaccination. With furunculosis, the success is attributed to the use of injectable vaccines containing adjuvants. Vaccines against virus infections, including infectious pancreatic necrosis, have also been used in commercial fish farming. Vaccines against several other bacterial and viral infections have been studied and found to be technically feasible. Pasteurellosis, streptococcosis (lactococcosis) and infections with iridoviruses are candidate diseases for control by immunoprophylaxis in the near future. The overall positive effect of vaccination in farmed fish is reduced mortality. However, for the future of the fish farming industry it is also important that vaccination contributes to a sustainable biological production with negligible consumption of antibiotics. A potential side-effect associated with injectable vaccines is local reactions in the peritoneal cavity. The paper presents recent developments in immunoprophylaxis of fish and some problems that should be addressed by the research community in the years to come.     

Muscle protein expression of pikeperches (Stizostedion lucioperca and S. volgense)

The purpose of the present study was to investigate the muscle protein expression in two pikeperches (Stizostedion lucioperca and S. volgense) through intra‐ and intermyomeric composition of white muscles. Using denaturing 10% sodium dodecylsulfate‐polyacrylamide gel electrophoresis, muscle protein expression was studied in relation to within‐ and between‐species morphological development, sex, maturity and age of pikeperches. Myosin, actin and troponin have a distinct role in the contraction and length tension of muscle fibers of these species. No obvious intramyomeric differences were found in the myosin heavy chain of both species. Myosin light chains (15–38 kDa) have different expression in different age groups. The muscle protein of the fingerling and adult S. lucioperca had high molecular weight (50 kDa) myosin in contrast to the other Percid species. The molecular weight of actins increased comparatively in low‐age‐group fish. ATP is stored in myosin and released to cause contraction when myosin comes in contact with actin of the experimental fish. Troponin regulates increasing concentration of light‐chain myosin in mature fish. Because troponin T has been implicated in the regulation of skeletal muscle kinetics, muscle contraction kinetics was predicted in different age groups. The muscle proteins of both sexes of these species have polymorphism in various age groups but have no difference in similar aged fish. No muscle protein dimorphism was found in these Percid species. The white muscle protein composition and contractile properties affect power production during fast, unsteady movement and swimming.     

Ophthalmic disease of fish.

Ocular disorders in fish are common and can occur as primary or secondary manifestations of systemic disease. Because most fish are sight feeders, visual impairments usually have fatal consequences for the individual fish and can lead to substantial economic losses for the client. This article encourages practitioners to perform a complete ocular examination as part of routine physical examinations in fish by providing a review of the functional anatomy of the teleostean eye, clinical work-up, ocular examination, ocular diseases, and treatment.     

Fine‐scale analysis of heavily invaded Italian freshwater fish assemblages

Inland waters are highly vulnerable to the introduction and spread of non‐native species, due to heavy human use of aquatic ecosystems and the natural linkages among streams and lakes. This is particularly noticeable in freshwater fish communities. To better evaluate how these communities are affected by non‐native species introductions, we conducted a fine‐scale analysis of the changes in Italian freshwater fish assemblages after species introduction. For this analysis, we collected information on fish species present in 44 basins. The present Italian freshwater fish fauna is composed of 48 native and 41 established introduced species, while a further 15 introduced species have been reported but are not yet considered naturalized. The changes in the fish assemblages mostly took place in the past 2 centuries and have increased recently, with nearly 60% of the species introduced in the past 3 decades. The number of species introduced per basin ranged from 0 to 35 (mean 10.85 ± 7.77 species/basin), and in 10 basins the number of species introduced is now equal to or even higher than the number of native species. In the past, introduced species mainly originated from America, but over the past three decades, an increase of introductions from other parts of Europe and Asia has been recorded. Our results show that basins already rich in native species tend to become even richer as a consequence of the establishment of introduced species. This confirms the trend toward a biotic homogenization of ecosystems even at a local scale, due to an increase in the human‐mediated spread of generalist species.     

Significance of two different ranking methods for studying fish fauna: a case study on the fish fauna from upper and middle portions from Mekong River

Studies on fish fauna should not only focus on fish composition and the comparison of fish composition among the study region and adjacent regions, but should also explore the origin and uniqueness of different taxa as well as the substitution of genera and species. In this study, the value of floristic presence method was modified and renamed the value of fish fauna presence (VFFP) method. The specific steps of the VFFP method and the traditional fish fauna analysis (TFFA) method were refined and standardized. Then, the VFFP and TFFA methods were applied to study the fish fauna of the upper and middle portions of the Mekong River basin. The results indicate that the TFFA method reflects the families (subfamilies) and genera that constitute the main body of fish in the studied river basin. The results of the VFFP method show which families (subfamilies) and genera are representative for the basin. Therefore, combining the TFFA and VFFP methods to analyze the composition of fish fauna can reflect the characteristics of fish fauna from different perspectives. The case study shows that the fish fauna of the Mekong River is a part of the fish fauna of Southeast Asia. Although it shows some similarities to the composition of South Asian fish fauna, it does not belong to the South Asian fish fauna as a whole, and is essentially different from the East Asian fish fauna. This study provides an objective, quantitative, and verifiable method for studying fish fauna.     

Piscirickettsiosis and piscirickettsiosis-like infections in fish: a review

Piscirickettsia salmonis was the first "rickettsia-like" bacteria to be recognized as a pathogenic agent of fish. Since the first reports of piscirickettsiosis emerged from Chile in the late 1980s, Piscirickettsia-like bacteria have been recognized with increasing frequency in a variety of fish species, from both fresh and saltwaters around the world. Although the first reported incidents of Piscirickettsia were in salmonids, Piscirickettsia-like bacteria are now being frequently associated with disease syndromes in non-salmonid fish. Mortalities have occurred in white seabass (Atactoscion noblis), black seabass (Dicentrarchus sp.), tilapia (Oreochromis, Tilapia and Sarotherodon spp.) and blue-eyed plecostomus (Panaque suttoni). Piscirickettsiosis and piscirickettsiosis-like diseases have affected aquaculture productivity, profitability, the species of fish compatible with commercial rearing, and transportation of fish from site to site. Piscirickettsiosis and syndromes caused by similar bacteria are an emerging disease complex that will increasingly inhibit fish production.     

Yersinia ruckeri,the causative agent of enteric redmouth disease in fish

Enteric redmouth disease (ERM) is a serious septicemic bacterial disease of salmonid fish species. It is caused by Yersinia ruckeri, a Gram-negative rod-shaped enterobacterium. It has a wide host range, broad geographical distribution, and causes significant economic losses in the fish aquaculture industry. The disease gets its name from the subcutaneous hemorrhages, it can cause at the corners of the mouth and in gums and tongue. Other clinical signs include exophthalmia, darkening of the skin, splenomegaly and inflammation of the lower intestine with accumulation of thick yellow fluid. The bacterium enters the fish via the secondary gill lamellae and from there it spreads to the blood and internal organs. Y. ruckeri can be detected by conventional biochemical, serological and molecular methods. Its genome is 3.7 Mb with 3406–3530 coding sequences. Several important virulence factors of Y. ruckeri have been discovered, including haemolyin YhlA and metalloprotease Yrp1. Both non-specific and specific immune responses of fish during the course of Y. ruckeri infection have been well characterized. Several methods of vaccination have been developed for controlling both biotype 1 and biotype 2 Y. ruckeri strains in fish. This review summarizes the current state of knowledge regarding enteric redmouth disease and Y. ruckeri: diagnosis, genome, virulence factors, interaction with the host immune responses, and the development of vaccines against this pathogen.     

Detection of Infectious Pancreatic Necrosis Virus from the Leeches Hemiclepsis marginata and Hirudo medicinalis

Leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV), and also may contain blood protozoa. In the present study, leeches were collected from water bodies located in Kurdistan province, Iran. The specimens were tested for IHNV, VHSV, and infectious pancreatic necrosis virus (IPNV) using the PCR method. The results showed that two different species of leeches, Hemiclepsis marginata and Hirudo medicinalis, were infected by IPNV among the seven species studied. The infected leeches were found in areas that were polluted with untreated sewage coming from upstream fish farms culturing Rainbow Trout Oncorhynchus mykiss. In addition, the fish at fish farms in the vicinity had been infected with IPNV 9 months previously. Our results showed that the virus causing infectious pancreatic necrosis is present in the leeches H. marginata and H. medicinalis, suggesting that leeches are a potential source of IPNV in fish farms.

Received October 14, 2015; accepted June 1, 2016 Published online September 29, 2016     

Host-dependent variations in the seasonal prevalence and intensity of heterophyid encysted metacercariae (Digenea: Heterophyidea) in brackish water fish in Egypt

The prevalence of heterophyid (Digenea: Heterophyidae) encysted metacercariae (EMC) in its second intermediate host, the fish Mugil spp. and Tilapia spp. was studied in a subtropical permanent Lake in northeastern Egypt. Seasonal changes in the occurrence of the EMC in different fish hosts were monitored in a longitudinal field survey lasting 12 months from June 2006 to May 2007. This study tested two hypotheses; (i) prevalence and intensity of heterophyid EMC fluctuate seasonally throughout the year and (ii) variation in the prevalence and intensity of heterophyid EMC is host-dependent. A total of 832 fish specimens comprising 5 species collected from Manzala Lake, northeastern Egypt were examined by artificial gastric juice digestion for EMC. All five species of brackish water fish examined were found to harbor the EMC of the family Heterophyidae in their muscles. The overall infection prevalence of EMC over 12 months was 23.2%. The adult flukes recovered from puppies experimentally infected with morphologically different metacercariae from different fish species were compatible with six species belong to five genera of Heterophyidae, namely, Heterophyes heterophyes, Heterophyes aequalis, Pygidiopsis genata, Phagicola sp., Haplorchis sp. and Stictodora sp. EMC of H. heterophyes were most abundant, detected in 56% of the total fish examined. P. genata was ranked second, followed by Phagicola sp., H. aequalis, Haplorchis sp., and Stictodora sp., respectively. Seasonal differences in infection were observed for all heterophyid species studied in all fish species examined. Heterophyid infections reached peak prevalences during the summer season 38.2% followed by spring 26.6% and autumn 19.3% seasons, whereas the lowest prevalence was recorded in the winter 8.7%. Intensity of heterophyid EMC followed the same seasonal pattern, being high during summer months and low in winter months. All fish species were infected with all the heterophyid digeneans, but with different prevalence. Possible reasons for these findings are discussed with reference to host, climatic and biotic factors.