The application of epidemiology in aquatic animal health -opportunities and challenges

ABSTRACT: Over recent years the growth in aquaculture, accompanied by the emergence of new and transboundary diseases, has stimulated epidemiological studies of aquatic animal diseases. Great potential exists for both observational and theoretical approaches to investigate the processes driving emergence but, to date, compared to terrestrial systems, relatively few studies exist in aquatic animals. Research using risk methods has assessed routes of introduction of aquatic animal pathogens to facilitate safe trade (e.g. import risk analyses) and support biosecurity. Epidemiological studies of risk factors for disease in aquaculture (most notably Atlantic salmon farming) have effectively supported control measures. Methods developed for terrestrial livestock diseases (e.g. risk-based surveillance) could improve the capacity of aquatic animal surveillance systems to detect disease incursions and emergence. The study of disease in wild populations presents many challenges and the judicious use of theoretical models offers some solutions. Models, parameterised from observational studies of host pathogen interactions, have been used to extrapolate estimates of impacts on the individual to the population level. These have proved effective in estimating the likely impact of parasite infections on wild salmonid populations in Switzerland and Canada (where the importance of farmed salmon as a reservoir of infection was investigated). A lack of data is often the key constraint in the application of new approaches to surveillance and modelling. The need for epidemiological approaches to protect aquatic animal health will inevitably increase in the face of the combined challenges of climate change, increasing anthropogenic pressures, limited water sources and the growth in aquaculture. TABLE OF CONTENTS: 1 Introduction 42 The development of aquatic epidemiology 73 Transboundary and emerging diseases 93.1 Import risk analysis (IRA) 103.2 Aquaculture and disease emergence 113.3 Climate change and disease emergence 133.4 Outbreak investigations 134 Surveillance and surveys 154.1 Investigation of disease prevalence 154.2 Developments in surveillance methodology 164.2.1 Risk-based surveillance and scenario tree modelling 164.2.2 Spatial and temporal analysis 164.3 Test validation 175 Spread, establishment and impact of pathogens 185.1 Identifying routes of spread 185.1.1 Ex-ante studies of disease spread 195.1.2 Ex-post observational studies 215.2 Identifying risk factors for disease establishment 235.3 Assessing impact at the population level 245.3.1 Recording mortality 245.3.2 Farm health and production records 265.3.3 Assessing the impact of disease in wild populations 276 Conclusions 317 Competing interests 328 Authors' contributions 329 Acknowledgements 3310 References 33.     

Epidemiological approach to aquatic animal health management: opportunities and challenges for developing countries to increase aquatic production through aquaculture

Aquaculture appears to have strongest potential to meet the increasing demands for aquatic products in most regions of the world. The world population is on the increase, as is the demand for aquatic food products. Production from capture fisheries at a global level is levelling off. Potential contributions from aquaculture to local food security, livelihoods and nutrition can be highly significant, especially in many remote and resource-poor rural areas. One of the major constraints to aquaculture production is the losses due to diseases. Over the decades, the sector has faced significant problems with disease outbreaks and epidemics which caused significant economic losses. The use of sound epidemiological principles and logical and science-based approach to identify and manage risks comprise two of the most important components of an effective biosecurity program. The maintenance of effective biosecurity in aquaculture is becoming more and more essential. There will be more demand for aquatic animal epidemiologists as well as epidemiological tools/resources in the region. The use of epidemiology will significantly improve health management, risk analysis and disease control. Although there are clear limitations and complications in the use of epidemiology for controlling aquatic animal pathogens, some positive results have recently emerged from a series of studies and trials to control diseases affecting the small-scale shrimp farming sector in southern India. This paper summarises the results of one such study which emphasizes the significant benefit of close collaboration with farmers, both individually and as groups, and capacity and awareness building among them and the importance of understanding the risk factors and implementing better management practices.     

Molecular epidemiological studies of veterinary arboviral encephalitides

Recent studies using molecular genetic approaches have made important contributions to our understanding of the epidemiology of veterinary arboviral encephalitides. Viruses utilizing avian enzootic hosts, such as Western equine encephalitis virus (WEEV) and North American Eastern equine encephalitis virus (EEEV), evolve as relatively few, highly conserved genotypes that extend over wide geographic regions; viruses utilizing mammalian hosts with more limited dispersal evolve within multiple genotypes, each geographically restricted. Similar findings have been reported for Australian alphaviruses. This difference may be related to vertebrate host relationships and the relative mobility of mammals and avians. Whereas EEEV and Venezualan equine encephalitis virus (VEEV) utilize small mammalian hosts in the tropics, most WEEV genotypes probably utilize avian hosts in both North and South America. The ability of mobile, infected avian hosts to disperse alphaviruses may result in continual mixing of virus populations, and thus limit diversification. This high degree of genetic conservation is also exhibited by EEE and Highlands J viruses in North America, where passerine birds serve as amplifying hosts in enzootic transmission foci. Most equine arboviral pathogens, including EEEV, WEEV and Japanese encephalitis virus (JEV), occur in a naturally virulent enzootic state and require only appropriate ecological conditions to cause epizootics and epidemics. However, VEE epizootics apparently require genetic changes to convert avirulent enzootic strains into distinct epizootic serotypes. All of these arboviruses have the potential to cause severe disease of veterinary and human health importance, and further molecular epidemiological studies will undoubtedly improve our ability to understand and control future emergence.     

The immunology of the bovine respiratory disease complex.

The bovine respiratory disease complex continues to be an economically important syndrome in an era when immunologic control is likely to become increasingly important. Recent studies have yielded a better understanding of the interaction, at the molecular level, of various pathogens with the bovine immune system. Improved challenge models for important viral pathogens such as bovine viral diarrhea virus and bovine respiratory syncytial virus have provided evidence of the efficacy of immune responses stimulated by vaccination. This article highlights recent advances in understanding of the role of the immune response in the pathogenesis and prophylaxis of bovine respiratory disease complex.     

Epidemiological studies on Salmonella and Campylobacter jejuni

An overview is given of investigations concerning the epidemiology of Salmonella, carried out at the National Institute of Public Health and Environmental Hygiene in Bilthoven, The Netherlands, during the last thirty years. It is made clear that Salmonella, because of its ubiquitous occurrence and its large variety in sero- and phage-types, is the organism of choice to study the epidemiological pathways of pathogens between man, animals and the environment. It is demonstrated that these are in fact the pathways of faecal contamination, and therefore have validity for a larger number of bacterial, and perhaps even parasitic and viral, micro-organisms. This last statement is illustrated by the presentation of studies on the epidemiology of Campylobacter jejuni.     

Human and porcine Taenia solium infections in Mozambique: identifying research priorities

The objective of this paper is to critically review and summarize available scientific and lay literature, and ongoing studies on human and porcine cysticercosis in Mozambique to identify knowledge gaps and direct immediate and long-term research efforts. Data on the spatial distribution and prevalence of the disease in human and swine populations are scarce and fragmented. Human serological studies have shown that 15-21% of apparently healthy adults were positive for cysticercosis antibodies or antigen, while in neuropsychiatric patients seroprevalence was as high as 51%. Slaughterhouse records indicate a countrywide occurrence of porcine cysticercosis, while studies have shown that 10-35% of pigs tested were seropositive for cysticercosis antibodies or antigen. Current research in Mozambique includes studies on the epidemiology, molecular biology, diagnosis and control of the disease. Future research efforts should be directed at better understanding the epidemiology of the disease in Mozambique, particularly risk factors for its occurrence and spread in human and swine populations, documenting the socio-economic impact of the disease, identifying critical control points and evaluating the feasibility and epidemiological impact of control measures and development of local level diagnostic tools for use in humans and swine.     

Effects of a Prophylactic Chloramine-T Treatment on Growth Performance and Condition Indices of Rainbow Trout

Abstract

Chloramine-T has been commonly used as a therapeutic agent for the treatment of bacterial gill disease and related phenomena in the salmonid aquaculture industry, and it is now commonly used as a prophylactic agent to prevent diseases in commercial salmonid hatcheries. Rainbow trout Oncorhynchus mykiss (average weight 98 g) were exposed to chloramine-T at 10 mg/L for 1 h twice weekly (a commonly used regime in the aquaculture industry) for 11 weeks. Fish were fed ad libitum without feed wastage throughout the 11 -week trial. Body weight, specific growth rate, feed conversion index, and appetite were assessed at the end of weeks 3, 6, 9, and 11. Chloramine-T treatment was not associated with either clinical disease or mortality. However, by the end of the trial, growth (based on body weight) of treated fish was significantly suppressed (δ = 7.3%) compared with control fish. Growth suppression was attributed to a significant reduction (δ = 9.3%) of feed conversion efficiency in treated fish. Based on specific growth rates, chloramine-T had an early negative effect on growth. The effect was diminished in later weeks (although not completely lost), which suggests some degree of compensation by the fish to the chemical agent.     

Epidemiological studies on Salmonella and Campylobacter jejuni

Summary

An overview is given of investigations concerning the epidemiology of Salmonella, carried out at the National Institute of Public Health and Environmental Hygiene in Bilthoven, The Netherlands, during the last thirty years. It is made clear that Salmonella, because of its ubiquitous occurrence and its large variety in sero‐ and phage‐types, is the organism of choice to study the epidemiological pathways of pathogens between man, animals and the environment. It is demonstrated that these are in fact the pathways of faecal contamination, and therefore have validity for a larger number of bacterial, and perhaps even parasitic and viral, micro‐organisms. This last statement is illustrated by the presentation of studies on the epidemiology of Campylobacter jejuni.     

Antagonistic effect of indigenous skin bacteria of brook charr (Salvelinus fontinalis) against Flavobacterium columnare and F. psychrophilum

Industrial fish production exposes fish to potentially stressful conditions, which in turn may induce infections by opportunistic pathogens. Probiotics appear to be a promising way to prevent opportunistic infections in aquaculture. In this study, we tested the inhibitory potential of endogenous bacterial communities found in the mucus of brook charr (Salvelinus fontinalis) against two major pathogens Flavobacterium columnare and Flavobacterium psychrophilum. Nine bacterial strains were isolated from brook charr skin mucus and tested for potential antagonistic activity. Results from both agar diffusion assays and broth co-culture assays showed the presence of antagonism. We identified seven bacterial strains, collected from unstressed fish, which exerted strong antagonism against F. psychrophilum and/or F. columnare. These strains were mixed and used to treat columnaris disease in an in vivo experiment in which four distinct fish families were tested. This treatment resulted in a decrease of mortality (54-86%) across fish families indicating that candidates from the host microbiota are potentially suitable for probiotic development. This would allow for the efficient (ability to adhere and colonize the host mucus) and durable management (antagonistic effect against pathogens which would be harmless for the host and safe for its environment) of opportunistic diseases in aquaculture.     

Streptococcus iniae: an aquatic pathogen of global veterinary significance and a challenging candidate for reliable vaccination

Streptococcus iniae has become one the most serious aquatic pathogens in the last decade causing high losses in farmed marine and freshwater finfish in warmer regions. Although first identified in 1976 from a captive Amazon freshwater dolphin, from which it derives its name, disease outbreaks had most likely been occurring for several decades in marine aquaculture in Japan. S. iniae is globally distributed throughout warm water finfish aquaculture. In common with other encapsulated beta-haemolytic streptococci and in direct contradiction to the phenomenal success story of bacterial vaccines in finfish aquaculture, control of S. iniae by vaccination has met with limited success. Thus, antibiotic usage is the current practice for reducing mortality and consequent economic loss. Vaccine failure appears to result in part from serotypic variation and, whilst 2 serotypes have been named, variation would appear to be more complex. S. iniae also has zoonotic potential, with human infections identified in the USA, Canada, and throughout Asia. In humans, infection is clearly opportunistic with all cases to date associated with direct infection of puncture wounds during preparation of contaminated fish, and generally in elderly or immunocompromised individuals. Significant progress has been made in terms of research into pathogenic mechanisms of S. iniae, with recent research elucidating the role of capsule in virulence for fish through antiopsonic activity. In light of this recent coverage in the literature, the present review centres on areas of direct veterinary interest including identification, epidemiology, therapy and prevention in farmed finfish. Clearly as the prevalence of S. iniae and associated economic losses continue to increase, further work towards developing a reliable vaccine is essential. This would appear to require a much better understanding of cell-surface variability amongst S. iniae isolates.     

Comparative susceptibility of Deschutes River, Oregon, Tubifex tubifex populations to Myxobolus cerebralis

Dams along the Deschutes River (DR) in central Oregon have blocked fish migration for over 40 years. Reestablishment of anadromous fish runs above the dams as part of a fish passage plan may introduce fish pathogens, such as Myxobolus cerebralis, the myxozoan parasite that causes salmonid whirling disease. This parasite is carried by adult salmon that stray into the DR system during their return to enzootic areas of the upper Columbia River basin, and it is now known to be established in at least one lower DR tributary. The life cycle of M. cerebralis involves two obligate hosts: a salmonid and the oligochaete worm Tubifex tubifex. To determine the likelihood of parasite establishment above the DR dams, we conducted benthic sediment surveys between 1999 and 2007 and found that T. tubifex had a patchy distribution and low relative abundance. Mitochondrial 16S ribosomal DNA gene analysis indicated that two lineages of T. tubifex (III and VI) were present both above and below the dams. Laboratory susceptibility studies to characterize differences in infection prevalence and parasite production between nine T. tubifex populations revealed that production varied considerably among exposed groups and was proportional to the number of lineage III worms present. Our results suggest that M. cerebralis could become established above the dams if infected fish are allowed passage into the upper DR system, but not all areas of the DR basin can be classified as having the same likelihood for parasite establishment, and the potential impact will be location dependent.     

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.     

Molecular epidemiology of antimicrobial resistance in veterinary medicine: where do we go?

Molecular epidemiology allows us to trace specific microorganisms and mobile genetic elements and to assess their epidemiological and evolutionary relationships. Examples of molecular epidemiology investigations in veterinary hospitals are discussed. They demonstrate the great similarities with the situation in human medicine and the potential usefulness of molecular epidemiology in our fight against antimicrobial resistance and nosocomial infections in veterinary hospitals. A broad knowledge of the diversity of antimicrobial resistance determinants in some major groups of pathogens and commensals from animals such as Enterobacteriaceae, Pasteurellaceae, enterococci and staphylococci is emerging. However, there are important gaps in this knowledge, which are discussed here. Many more molecular epidemiology studies will be necessary to understand and follow the evolution of the problem in veterinary medicine and agriculture on a global scale. To be able to build useful surveillance programs and reliable epidemiological models, and to identify critical intervention points, we need to improve our understanding of antimicrobial resistance at the animal and farm levels. Studies assessing the dynamics of bacterial populations and of resistance determinants at these levels are desperately needed. Understanding the relationships between antimicrobial resistance, colonization factors, and virulence also represents a major issue for which molecular epidemiology investigations will be needed.     

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.     

The role of wildlife in transboundary animal diseases

This paper identifies some of the more important diseases at the wildlife-livestock interface and the role wildlife plays in disease transmission. Domestic livestock, wildlife and humans share many similar pathogens. Pathogens of wild or domestic animal origin that can cause infections in humans are known as zoonotic organisms and the converse are termed as anthroponotic organisms. Seventy-seven percent of livestock pathogens and 91% of domestic carnivore pathogens are known to infect multiple hosts, including wildlife. Understanding this group of pathogens is critical to public health safety, because they infect a wide range of hosts and are most likely to emerge as novel causes of infection in humans and domestic animals. Diseases at the wildlife-livestock interface, particularly those that are zoonotic, must be an area of focus for public health programs and surveillance for emerging infectious diseases. Additionally, understanding wildlife and their role is a vital part of understanding the epidemiology and ecology of diseases. To do this, a multi-faceted approach combining capacity building and training, wildlife disease surveillance, wildlife-livestock interface and disease ecology studies, data and information sharing and outbreak investigation are needed.     

Emerging pathologies in aquaculture: effects on production and food safety

Infectious diseases represent a limiting factor for the further development of Italian aquaculture. The recent introduction and spreading of new pathogens, along with the global climatic change, has contributed to a considerable decrease in trout production. Emerging pathologies in rainbow trout culture include viral diseases, e.g. infectious haematopoietic necrosis (IHN), bacterial diseases, such as lactococcosis and visceral flavobacteriosis, and parasitical diseases, e.g. proliferative kidney disease (PKD). Higher mortality rates in trout fry and fingerlings are generally induced by visceral flavobacteriosis and IHN, while the main losses in large trout during the warm season are due to lactococcosis and PKD. Mariculture has at present a better sanitary status compared to trout culture, but a rapid dissemination of pathogens, including zoonosis agents, is envisaged also for seabass and seabream. Emerging pathologies in sea bass include VNN, pseudotuberculosis, streptococcosis and tuberculosis. Seabream is much more resistant and is mainly affected by novel Vibrio infections and enteromyxidiosis. A good sanitary management of fish farms is essential for avoiding or limiting losses caused by emerging pathologies. Transmission of zoonosis agents to man, through the consumption of cultured fish, is very remote in Italy. On the contrary, transmission of Streptococcus iniae, Vibrio vulnificus and Mycobacterium marinum by means of improper manipulation of infected fish, could represent a potential hazard for fish farmers and fish processors, as well as for people preparing fish meals.     

Bacterial zoonoses of fishes: A review and appraisal of evidence for linkages between fish and human infections

Human contact with and consumption of fishes presents hazards from a range of bacterial zoonotic infections. Whereas many bacterial pathogens have been presented as fish-borne zoonoses on the basis of epidemiological and phenotypic evidence, genetic identity between fish and human isolates is not frequently examined or does not provide support for transmission between these hosts. In order to accurately assess the zoonotic risk from exposure to fishes in the context of aquaculture, wild fisheries and ornamental aquaria, it is important to critically examine evidence of linkages between bacteria infecting fishes and humans. This article reviews bacteria typically presented as fish-borne zoonoses, and examines the current strength of evidence for this classification. Of bacteria generally described as fish-borne zoonoses, only Mycobacterium spp., Streptococcus iniae, Clostridium botulinum, and Vibrio vulnificus appear to be well-supported as zoonoses in the strict sense. Erysipelothrix rhusiopathiae, while transmissible from fishes to humans, does not cause disease in fishes and is therefore excluded from the list. Some epidemiological and/or molecular linkages have been made between other bacteria infecting both fishes and humans, but more work is needed to elucidate routes of transmission and the identity of these pathogens in their respective hosts at the genomic level.     

Tick-borne infections in dogs-an emerging infectious threat

Many viral, bacterial and parasitic pathogens have been associated with tick transmission, including several recently identified pathogens in both humans and domestic animals, especially dogs. The emergence in dogs of these tick-borne infections has a multi factorial origin. Better animal care, better diagnostic tools, and a broader distribution of the vectors in favorable habitats through population migrations including travel with owned pets, translocation or commercial trade of pet dogs, are some of the factors contributing to the emergence and recognition of these new pathogens. The present review focuses on the recent epidemiological studies which support the emergence or re-emergence of tick-borne pathogens in dogs around the world, as well as give some insight on newly recognized potentially tick-borne pathogens, such as Bartonella infections.     

Comparison of Selected Nonlethal Samples from Adult Steelhead for Detection of Infectious Hematopoietic Necrosis Virus Using Cell Culture

Abstract

Nonlethal sampling techniques have previously been evaluated for detection of a variety of viral salmonid pathogens. However, many of these studies have used molecular assays in lieu of widely accepted cell culture techniques to evaluate the sampled tissues. Samples were collected from female steelhead Oncorhynchus mykiss broodstock using three potential nonlethal sampling methods (mucus/skin scrape, pectoral fin clip, and gill tissue biopsy) and evaluated for the presence of infectious hematopoietic necrosis virus (IHNV) via cell culture techniques. The results were compared with those from samples collected using a standard lethal sampling method (pooled anterior kidney and spleen tissues) applied to the same fish. Of the three nonlethal sampling techniques that were evaluated, fin clipping was the easiest and least invasive method. Furthermore, fin tissue was as sensitive as or more sensitive than kidney/spleen tissue for detecting IHNV in this population of fish. However, with the exception of gill tissue, the nonlethal samples did not appear to be appropriate surrogates for lethally collected tissues with regard to identifying an active infection in a particular fish. Nevertheless, nonlethal sampling coupled with cell culture appears to be suitable for helping to define the IHNV status of a steelhead population.

Received July 27, 2016; accepted December 11, 2016 Published online March 21, 2017     

A framework for understanding the potential for emerging diseases in aquaculture

Numerous diseases have emerged as serious economic or ecological problems in aquaculture species. The combination of factors behind the emergence of each disease is unique, but various common factors are apparent. We combine risk-analysis methods and virulence theory with historical examples (mainly from salmonid production) to identify key disease-emergence risk factors. Diseases have emerged through pathogen exchange with wild populations, evolution from non-pathogenic micro-organisms and anthropogenic transfer of stocks. Aquacultural practices frequently result in high population densities and other stresses (such as intercurrent disease) which increase the risk of infection establishment and spread. As aquaculture expands and new species are farmed, diseases will continue to emerge and affect both wild and farmed fish adversely. The rate and extent of emergence can be reduced by the application of biosecurity programmes designed to mitigate the risk factors for disease emergence.