Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Occurrence of Mycobacterium spp. in ornamental fish in Italy

The occurrence of Mycobacterium spp. in freshwater and marine ornamental fish was studied in Italy from June 2002 to May 2005. Two surveys were carried out, one of aquarium fish sent to the Laboratory for diagnosis, and the other of prevalence of infection by mycobacteria in ornamental fish imported into Italy. Bacterial isolation was carried out from the spleen, kidney and liver, and the isolates were subsequently identified by biochemical tests. In the first survey, 387 fish were examined and Mycobacterium spp. were isolated from 181 (46.8%) fish. In the second survey 127 batches of ornamental fish from different countries were examined. Mycobacterium spp. were isolated from 38 (29.9%) batches. The following species were found: M. fortuitum, M. peregrinum, M. chelonae, M. abscessus, M. marinum, M. gordonae, M. nonchromogenicum and M. interjectum. There was a high prevalence of infection independent of the presence of macroscopic lesions. Mycobacterium fortuitum and M. chelonae were more prevalent than M. marinum in the samples examined.     

United States of America Trade in Ornamental Fish

Ornamental fish production is among the leading cash crops of the United States of America aquaculture economy, and retail value of the fish trade is worth approximately US$l,000 million. However, few studies exist to document this trade. Using import and export documents we report trends in total values for the U.S. trade in ornamental fish. Also, we determined the number and value of the most commonly imported ornamental fish. In 1992, approximately 201 million fish valued at $44.7 million were imported into the U.S. The port of Los Angeles handled 39% of all trade activity, Miami 22%, New York 16%, Tampa 6%, and Honolulu 6%. Freshwater fish accounted for approximately 96% of the total volume and 80% of the value of the imports. Most freshwater ornamental fish were farm-raised and imported from Southeast Asia. Most U.S. exports of ornamental fish were cultured in the state of Florida. Although saltwater fish had a high market value (20% of the declared value of the imports), the volume of these fish was only 4%. Most imported and exported saltwater ornamental fish were collected from the wild. Of the 1,539 species declared as ornamental fish, 32 species dominated the trade. These were all of freshwater origin. The guppy Poecilia reticulala and neon tetra Paracheirodon innesi were the most popular ornamental fish kept in U.S. households. The average prices paid for imported ornamental freshwater fish were 45 cents for egg layers and 22 cents for live bearers. The results of this study document the importance of the ornamental fish industry and identifies the most valuable species in the trade for potential domestic culture and protection in the wild.     

Iridovirus infections in finfish – critical review with emphasis on ranaviruses

Viruses in three genera of the family Iridoviridae (iridoviruses) affect finfish. Ranaviruses and megalocytiviruses are recently emerged pathogens. Both cause severe systemic disease, occur globally and affect a diversity of hosts. In contrast, lymphocystiviruses cause superficial lesions and rarely cause economic loss. The ranavirus epizootic haematopoietic necrosis virus (EHNV) from Australia was the first iridovirus to cause epizootic mortality in finfish. Like other ranaviruses, it lacks host specificity. A distinct but closely related virus, European catfish virus, occurs in finfish in Europe, while very similar ranaviruses occur in amphibians in Europe, Asia, Australia, North America and South America. These viruses can be distinguished from one another by conserved differences in the sequence of the major capsid protein gene, which informs policies of the World Organisation for Animal Health to minimize transboundary spread of these agents. However, limited epidemiological information and variations in disease expression create difficulties for design of sampling strategies for surveillance. There is still uncertainty surrounding the taxonomy of some putative ranaviruses such as Singapore grouper iridovirus and Santee‐Cooper ranavirus, both of which cause serious disease in fish, and confusion continues with diseases caused by megalocytiviruses. In this review, aspects of the agents and diseases caused by ranaviruses are contrasted with those due to megalocytiviruses to promote accurate diagnosis and characterization of the agents responsible. Ranavirus epizootics in amphibians are also discussed because of possible links with finfish and common anthropogenic mechanisms of spread. The source of the global epizootic of disease caused by systemic iridoviruses in finfish and amphibians is uncertain, but three possibilities are discussed: trade in food fish, trade in ornamental fish, reptiles and amphibians and emergence from unknown reservoir hosts associated with environmental change.     

Health Status of Ornamental Freshwater Fishes Imported to South Africa: A Pilot Study

Keeping fish is a popular pastime in South Africa and the majority of ornamental fish are imported. A pilot study was initiated to examine the health status of ornamental freshwater fishes imported to South Africa. Four groups of thirty fish each were examined for the presence of external parasites, and processed for virus isolation and for bacterial and mycobacterial culture. The groups consisted of goldfish (Carassius auratus), koi (Cyprinus carpio), guppies (Poecilia reticulata) and cardinal tetras (Cheirodon axelrodi). Mycobacterium fortuitum was isolated from the goldfish and koi. No other significant bacteria were isolated and virus culture was negative for all groups. Skin scrapings and wet gill preparations were made to detect external parasites. Parasites were identified from fixed material. External parasites included Trichodina mutabilis, Ichthyophthirius multifiliis, ciliophorans of the genus Tetrahymena, and monogeneans belonging to the genera Dactylogyrus and Gyrodactylus. This is the first report of Trichodina mutabilis in South Africa. Diseases imported with ornamental fish pose a risk to both indigenous fish populations and the aquaculture industry. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of real-time PCR assays for detection of megalocytiviruses in imported ornamental fish

Megalocytiviruses have been associated globally with severe systemic disease and economic loss in farmed food fish and ornamental fish. The viruses have been spread internationally by translocation of live fish. In New Zealand, megalocytiviruses are regarded as exotic. A potential pathway for introduction has been identified, namely imported ornamental fish. In the present study, real‐time PCR assays were developed for detection of megalocytiviruses using a conserved major capsid protein gene. A SYBR green assay was developed to target all known megalocytiviruses. A second real‐time PCR assay using a molecular beacon was developed to specifically target gourami, Trichogaster trichopterus, iridovirus, a species of iridovirus previously linked to ornamental fish imports in Australia. The analytical sensitivity for the SYBR green and molecular beacon assays were 10 and 100 fg, respectively. The analytical specificity of the real‐time PCR assays determined using genomic DNA templates from three target viruses, 12 non‐target viruses and 25 aquatic bacterial species were 100%. The intra‐run and inter‐run coefficients of variation of both assays were <5%. The real‐time PCR assays developed in this study provide rapid, sensitive, and specific detection of megalocytiviruses and gourami iridovirus.     

Identification of imported Chirimen at the genus level by a direct sequencing method using mitochondrial partial 16S rDNA region

ABSTRACT:   Quality control of imported fish products is a constant challenge for Customs officials. To help officials access the quality and integrity of imported fish products, and to enhance the effectiveness and accuracy of product screening, a method was developed to identify boiled and dried larval and juvenile sardines (Chirimen) at the genus level by direct sequencing using a partial sequence (606 bp) of mitochondrial 16S rDNA. A phylogenetic tree was constructed, based on nucleotide sequences of seven kinds of Chirimen imported from four countries with 16 standard samples belonging to the orders Clupeiformes and Salmoniformes. To identify the fish genera in imported Chirimen, bootstrap values and branch lengths of the phylogenetic tree as well as genetic distances were used. The data revealed that Chirimen imported from China and Korea contains species of the genus Engraulis . Regarding other imported Chirimen, the least that could be verified was that the sardine ingredients do not belong to regulated genera.     

Detection and characterization of viruses of the genus Megalocytivirus in ornamental fish imported into an Australian border quarantine premises: an emerging risk to national biosecurity

This report documents an emerging trend of identification of Megalocytivirus-like inclusions in a range of ornamental fish species intercepted during quarantine detention at the Australian border. From September 2012 to February 2013, 5 species of fish that had suffered mortality levels in excess of 25% whilst in the post-entry quarantine and had Megalocytivirus-like inclusion bodies in histological sections were examined by PCR. The fish had been imported from Singapore, Malaysia and Sri Lanka. Ninety-seven of 111 individual fish from affected tanks of fish tested were positive for the presence of Megalocytivirus by PCR. Sequence analysis of representative PCR products revealed an identical sequence of 621 bp in all cases which was identical to a previously characterized Megalocytivirus (Sabah/RAA1/2012 strain BMGIV48). Phylogenetic analysis of available Megalocytivirus major capsid protein (MCP) sequences confirmed the existence of 3 major clades of Megalocytivirus. The virus detected in this study was identified as a member of Genotype II. The broad host range and pathogenicity of megalocytiviruses, coupled to the documented spread of ornamental fish into the environment, render this a significant and emerging biosecurity threat to Australia.     

Marine ornamental fish imports in the European Union: an economic perspective

The marine ornamental fish trade (MOFT) is a worldwide industry that targets a remarkable quantity and diversity of reef fish species and provides an important source of revenue for exporting countries, particularly developing nations in Southeast Asia. Here we provide, for the first time, an economic assessment of the MOFT to the European Union (EU), one of the key players of the marine aquarium trade, along with the USA and Japan. Data from EUROSTAT (the European Statistical System) was analysed for the MOFT between 2000 and 2011. Results show that this industry was not affected by the economic crisis and accounted for a total of 135 million Euros during the study period. Both the UK and Indonesia have been reinforcing their importance as leading importer and exporter countries, respectively. The long‐term temporal pattern of the MOFT by EU and non‐EU countries is described, as well as its diversity and evenness between importing and exporting countries. The present economic assessment also identifies the key players of this industry, and ultimately contributes to management and conservation practices that may enhance the sustainable use of world's coral reefs.     

Detection and molecular characterization of infectious spleen and kidney necrosis virus from major ornamental fish breeding states in Peninsular Malaysia

‘Gold standard’ OIE reference PCR assay was utilized to detect the presence of infectious spleen and kidney necrosis virus (ISKNV) in freshwater ornamental fish from Malaysia. From total of 210 ornamental fish samples representing 14 species, ISKNV was detected in 36 samples representing 5 fish species. All positive cases did not show any clinical signs of ISKNV. Three restriction enzymes analyses showed that the fish were infected by identical strains of the same virus species within Megalocytivirus genus. Major capsid protein (MCP) genes of 10 ISKNV strains were sequenced and compared with 9 other reference nucleotide sequences acquired from GenBank. Sequence analysis of MCP gene showed that all strains detected in this study were closely related to the reference ISKNV with nucleotide sequence identity that was ranging from 99.8% to 100%. In addition, phylogenetic analysis of MCP gene revealed that viruses from genus Megalocytivirus can be divided into three genotypes: genotype 1 include reference ISKNV and all other strains that were detected in this study, genotype 2 include viruses closely related to red sea bream iridovirus (RSIV), and genotype 3 include viruses closely related turbot reddish body iridovirus (TRBIV).     

Fish movements: the introduction pathway for topmouth gudgeon Pseudorasbora parva and other non‐native fishes in the UK

• 1. The contamination of fish consignments (for stocking or aquaculture) is a major pathway by which non‐native organisms, including fish, are introduced to new areas. One of the best examples of this is the topmouth gudgeon Pseudorasbora parva, which was accidentally imported into Romania and then throughout Europe in consignments of Asian carp species.
• 2. The introduction and spread of topmouth gudgeon in the UK has been linked to imports and movements of the ornamental variety (golden orfe) of ide Leuciscus idus. To examine this hypothesis, relationships between authorized movements of both native and non‐native fish species (in particular ide) and the occurrence in England of topmouth gudgeon were tested at the 10×10 km scale.
• 3. Topmouth gudgeon occurrence in the wild was significantly correlated with the trajectories of movements of ornamental fish species (ide/orfe, sunbleak Leucaspius delineatus) as well as a few non‐ornamental fish species (European catfish Silurus glanis, Atlantic salmon Salmo salar and grass carp Ctenopharyngodon idella).
• 4. These results highlight the mechanism by which non‐native fish species disperse from the point of first introduction, and especially that movements of fish within the country represent an important mechanism for accidental introductions of non‐native species. © Crown copyright 2010. Reproduced with the permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.

     

An understated danger: Antimicrobial resistance in aquaculture and pet fish in Switzerland,a retrospective study from 2000 to 2017

Aquaculture is a rapidly growing field of food production. However, morbidity and mortality are higher in aquaculture species than in domestic animals. Bacterial diseases are a leading cause of farmed fish morbidity and are often treated with antimicrobials. Since most Swiss fish farms release effluents directly into surface water without treatment and since aquaculture fish are consumed by humans, antimicrobial resistance (AMR) and multi-resistance in aquaculture fish are important for environmental and public health. In this study, AMR tests for 14 antimicrobials were performed on 1,448 isolates from 1,134 diagnostic laboratory submissions from farmed and ornamental fish submissions for the period from 2000 to 2017. Amoxicillin, gentamycin and norfloxacin had the lowest proportion of resistant samples. However, AMR was highly variable over time. Resistance proportions were higher in: (a) ornamental fish compared with farmed fish, (b) fish from recirculation systems compared with those from other farming systems and (c) isolates originating from skin compared with those originating from inner organs. Multiple resistances were common. The results of this study provide useful data for Swiss fish veterinarians and some interesting hypotheses about risk factors for AMR in aquaculture and pet fish in Switzerland. However, further research is needed to define risk factors.     

中国水产苗种进口特征及其对养殖产业发展的影响分析——基于2013—2018年的数据

水产种业是水产养殖业的战略性、基础性核心产业,水产苗种引进是促进水产养殖业绿色发展的一个重要途径。中国水产苗种引进品种繁多,水产观赏动物成为新宠,通过分析2013—2018年水产苗种进口贸易数据,其主要特征为:进口额呈下降趋势,进口种类涉及24种,部分种类的进口频率达到100%,用于水产养殖的苗种和受精鱼卵进口额分别为14061.19×104美元和632.02×104美元。从进口额数据来看,小虾及对虾、鳗鱼、受精鱼卵及其他鱼种等主要进口苗种种类对水产养殖业发展的影响较大,其中鳗鱼增养殖业对进口鳗苗的依赖程度较高,平均依赖度为50.86%。提出构建养殖鱼类苗种进口预警机制,谋划养殖鱼类品种的替代策略,提高对虾苗种控制能力,做好水产苗种进口质量安全防范等建议,以保障水产养殖产业健康稳定发展。     

Comparison in taste and extractive components of boiled dorsal muscle and broth from half-smooth golden puffer Lagocephalus spadiceus caught in Japan with those of the same fish imported

The taste and extractive components of boiled dorsal muscle and broth prepared from half-smooth golden puffer Lagocephalus spadiceus caught in Japan and from those imported from China were compared. In the sensory test, the first taste, elasticity, and saltiness of boiled muscle from Japanese (domestic) fish were higher than those of the imported fish, whereas the orthonasal fishy smell of the imported fish was higher than that of the domestic fish. The first taste, aftertaste, retronasal fishy smell, sweetness, saltiness, and umami of the broth prepared from dorsal muscle of the domestic fish were higher than those of the imported fish, whereas orthonasal fishy smell and bitterness of the imported fish were higher than those of the domestic fish. Most panelists preferred the overall taste of the domestic fish to that of the imported fish. The concentration of trimethylamine in the trichloroacetic acid extracts from boiled muscle and broth of the imported fish was higher than that of the domestic fish, suggesting that this substance contributes to the orthonasal fishy smell of the imported fish.     

Development of real-time PCR assays for the detection and differentiation of Australian and European ranaviruses

Serious systemic disease in fish and amphibians is associated with the ranaviruses, epizootic haematopoietic necrosis virus (EHNV) and Bohle iridovirus (BIV) in Australia, and European sheatfish virus (ESV) and European catfish virus (ECV) in Europe. EHNV, ESV and ECV are recognized causative agents of the OIE (Office International des Epizooties) notifiable systemic necrotizing iridovirus syndrome and are currently identified by protein-based assays, none of which are able to rapidly identify the specific agents. The aim of this study was to develop TaqMan real-time PCR assays that differentiated these viruses using nucleotide sequence variation in two ranavirus genes. A conserved probe representing 100% sequence homology was used as a reference for virus-specific probes. The virus-specific probes produced a similar signal level to the conserved probe while those probes binding to non-target viral DNA produced an altered fluorescent curve. The pattern of probe binding was characteristic for each virus. Sensitivity, specificity and dynamic range of the assay were assessed. The test is currently useful as a research and initial screening tool, with the potential to become a sensitive and specific method for detection and differentiation of ranaviruses with further development.     

Economic,Regulatory, and Technological Barriers to Entry into the Florida Aquaculture Industry

A mail survey was conducted in 1991 to identify barriers-to-entry into the Florida aquaculture industry. The survey was administered to all 586 individuals on the Florida Agricultural Statistics Service aquaculture industry participant mailing list. Two hundred twenty of the 227 returned questionnaires were complete. Of these, 175 respondents currently owned an aquaculture business; 141 reported producing only one species (defined in this study to be the general collection of species such as ornamental fish or aquatic plants rather than specific species within these groups), 29 reported producing two different species, and four reported producing three or more species. In order to analyze the data by species; only those aquaculturists producing single species were included in the final analyses. The survey results showed that economic factors (financial and marketing) presented the greatest barriers-to=entry into the Florida aquaculture industry, while regulatory barriers were relatively less problematic. Barriers were found to be highest for catfish culturists, followed by ornamental fish, bivalves, aquatic plants and alligators.     

Effects of a probiotic, protexin, on the growth performance and hematological parameters in the Oscar (Astronotus ocellatus) fingerlings

This study examined the effects of a probiotic, protexin, on the growth performance and hematological parameters in an ornamental fish, the Oscar Astronotus ocellatus fingerlings. A completely randomized experimental design was applied with three experimental diets each with three replicates. A commercial food, BioMar, was supplemented with protexin at levels of 0.15, 0.5, and 1.0 g kg(-1) dry food and fed three times a day for 60 days. The control diet was prepared with no protexin supplementation. The experimental fish were biometried every 15 days to compare their growth rates at each treatment. For hematological assays, blood samples were prepared every 30 days to measure such parameters as red and white blood cells, hemoglobin, hematocrit, and percentages of lymphocytes, monocytes, neutrophiles, basophiles, and eosinophiles. Based on the results, the fingerlings fed a 0.15 g kg(-1) supplemented food were significantly different from the fish in the other treatments and in the control, with the highest mean of both final weight (35.07 ± 1.19) and body weight gain (30.17 ± 1.08). Significant differences in both hemoglobin concentration and mean red and white blood cells were found between the experimental groups and the control within 2 months. The highest hemoglobin concentration and also red and white blood cells was observed in the fish-fed 0.15 dietary protexin in both months. The results of this study show that the probiotic, protexin, at a level of 0.15 g kg(-1) dry food could have measurable effects on the growth and hematological parameters in the Oscar A. ocellatus fingerlings.     

Koi herpesvirus: distribution and prospects for control in England and Wales

Koi herpesvirus (KHV) causes a highly virulent disease affecting carp, Cyprinus carpio L., and poses a serious socio‐economic threat to the UK carp industry. This study aimed to determine the geographic distribution and prevalence of KHV exposed fish in England and Wales through ELISA antibody testing. Only three of the 82 farms sampled produced positive results, suggesting fish farms provide a relatively safe source of fish. Of the 71 ‘high‐risk’ fisheries tested, 26 were positive. All eight geographic areas within England and Wales studied had at least one KHV positive site. Twelve consignments of imported koi carp from seven S.E. Asian countries were tested for KHV antibody. Six consignments from six different countries were positive. Although a high proportion of consignments were positive, the results indicate that lower risk stocks of fish exist that could be sourced by the ornamental carp sector. The study provides evidence that KHV is widespread and prevalent in ‘high‐risk’ fisheries. There are, however, prospects for controlling KHV as English and Welsh farms appear to be relatively free of the virus, and in most cases fish are not moved from fisheries to other waters.