Antibiotic use during the intracoelomic implantation of electronic tags into fish

The use of antibiotics, in particular, the use of a single dose of antibiotics during electronic tag implantation is of unproven value, and carries with it the potential for the development of antibiotic resistance in bacteria and the alteration of the immune response of the fish. Antibiotic use during electronic tag implantation must conform to relevant drug laws and regulations in the country where work is being done, including the requirements for withdrawal times before human consumption is a possibility. Currently, the choice of antibiotics (most often tetracycline or oxytetracycline) and the use of a single dose of the drug are decisions made without knowledge of the basic need for antibiotic usage and of the bacteria involved in infections that occur following electronic tag implantation. Correct perioperative use of an antibiotic is to apply the drug to the animal before surgery begins, to assure serum and tissue levels of the drug are adequate before the incision is made. However, the most common perioperative application of antibiotics during implantation of an electronic tag is to delay the administration of the drug, injecting it into the coelom after the electronic tag is inserted, just prior to closure of the incision. There is little empirical evidence that the present application of antibiotics in fish being implanted with electronic tags is of value. Improvements should first be made to surgical techniques, especially the use of aseptic techniques and sterilized instruments and electronic tags, before resorting to antibiotics.     

Training considerations for the intracoelomic implantation of electronic tags in fish with a summary of common surgical errors

Training is a fundamental part of all scientific and technical disciplines. This is particularly true for all types of surgeons. For surgical procedures, a number of skills are necessary to reduce mistakes. Trainees must learn an extensive yet standardized set of problem-solving and technical skills to handle challenges as they arise. There are currently no guidelines or consistent training methods for those intending to implant electronic tags in fish; this is surprising, considering documented cases of negative consequences of fish surgeries and information from studies having empirically tested fish surgical techniques. Learning how to do fish surgery once is insufficient for ensuring the maintenance or improvement of surgical skill. Assessment of surgical skills is rarely incorporated into training, and is needed. Evaluation provides useful feedback that guides future learning, fosters habits of self-reflection and self-remediation, and promotes access to advanced training. Veterinary professionals should be involved in aspects of training to monitor basic surgical principles. We identified attributes related to knowledge, understanding, and skill that surgeons must demonstrate prior to performing fish surgery including a “hands-on” assessment using live fish. Included is a summary of common problems encountered by fish surgeons. We conclude by presenting core competencies that should be required as well as outlining a 3-day curriculum for training surgeons to conduct intracoelomic implantation of electronic tags. This curriculum could be offered through professional fisheries societies as professional development courses.     

Surgical implantation of transmitters into fish

Although the Animal Welfare Act does not cover poikilotherms, individual institutions and policies and legal requirements other than the Animal Welfare Act (e.g., the US Public Health Service and the Interagency Research Animal Committee's Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training) require the review of projects involving fish by institutional animal care and use committees (IACUCs). IACUCs may, however, lack the knowledge and experience to evaluate fish projects judiciously, especially when the projects are in field settings. Surgeries involving implantation of transmitters and other instruments into the coelom, which now comprise a very common research tool in the study of free-ranging fishes, are examples of surgeries that use a broad spectrum of surgical and anesthetic techniques, some of which would not be considered acceptable for similar work on mammals. IACUCs should apply the standards they would expect to be used for surgeries on homeotherms to surgeries on fish. Surgeons should be carefully trained and experienced. Surgical instruments and transmitters should be sterile. Regulations and laws on the use of drugs in animals should be followed, particularly those concerned with anesthetics and antibiotics used on free-ranging fish. Exceptions to surgical procedures should be made only when circumstances are extreme enough to warrant the use of less than optimal procedures.     

Evaluation of water entry into the coelom and different levels of aseptic technique during surgical implantation of electronic tags in freshwater fish

We reviewed the literature in an attempt to determine the importance of aseptic technique when implanting electronic tags in fish. Given that there was negligible information on this topic we embarked on a study where bluegill (Lepomis macrochirus) were used as a model to investigate the effects of different aseptic surgical techniques for the intracoelomic implantation of electronic tags in fish. First we tested the effects of water entry into the incision using five treatments: lake, distilled and saline water introduced into the incision, water-free controls, and non-surgery controls. For fish in the water treatments, 1 mL of the sample was introduced into their coelom prior to incision closure. Fish were held for 10 days to monitor survival and at the end of the study, the survivors were blood sampled and euthanized to evaluate condition and health using the health assessment index. In a second experiment, four aseptic treatments were used: non-sterile, field-based, high-grade sterility, and non-surgery controls and fish were monitored as in the first experiment. For both experiments, no differences in physiological status, health or mortality were noted among treatment groups. However, in the aseptic techniques experiment, surgical times were approximately twice as long for fish in the sterile treatment as compared to other groups and the costs of surgical supplies was greater than that of the less-sterile treatments. Although we failed to document any benefit of keeping water out of the incision or using aseptic technique for bluegill, in other situations and for other species, such approaches may be important. As such, we encourage fish surgeons practicing intracoelomic implants to attempt to prevent water entry into the coelom. We also encourage, at least some level of infection control (e.g., non-sterile gloves, clean tags and surgical tools) consistent with good veterinary practices to maintain the welfare status of tagged fish and ensure that the data from tagged fish representative of untagged conspecifics. However, the most prudent and ethical approach would be to work with veterinarians to incorporate formal sterilization procedures, equipment (e.g., sterile gloves) and aseptic technique into field surgical techniques.     

Holding of juvenile salmonids for surgical implantation of electronic tags: a review and recommendations

Many telemetry-based studies require that fish be sampled from the wild and then held both prior to and after the implantation of an electronic tag. However, the effects of such holding (or the lack thereof) have yet to be studied intensively. Pre-surgical holding often occurs to facilitate logistical needs of research projects and as an attempt to minimize negative physiological effects due to capture and handling stress. Further, post-surgical holding time and conditions greatly influence the physiological state of fish prior to being returned to the wild. This paper reviews pertinent studies pertaining to the effects of surgical holding on the behavior, physiology, and survival of fishes, with particular emphasis on juvenile salmonids. The effects of individual aspects of surgical holding such as handling, water quality, light conditions, holding density, metabolic scope, and duration of holding are reviewed. Recommendations regarding certain aspects of surgical holding are offered with a goal of reducing bias related to the surgical process.     

An introduction to the practical and ethical perspectives on the need to advance and standardize the intracoelomic surgical implantation of electronic tags in fish

The intracoelomic surgical implantation of electronic tags (including radio and acoustic telemetry transmitters, passive integrated transponders and archival biologgers) is frequently used for conducting studies on fish. Electronic tagging studies provide information on the spatial ecology, behavior and survival of fish in marine and freshwater systems. However, any surgical procedure, particularly one where a laparotomy is performed and the coelomic cavity is opened, has the potential to alter the survival, behavior or condition of the animal which can impair welfare and introduce bias. Given that management, regulatory and conservation decisions are based on the assumption that fish implanted with electronic tags have similar fates and behavior relative to untagged conspecifics, it is critical to ensure that best surgical practices are being used. Also, the current lack of standardized surgical procedures and reporting of specific methodological details precludes cross-study and cross-year analyses which would further progress the field of fisheries science. This compilation of papers seeks to identify the best practices for the entire intracoelomic tagging procedure including pre- and post-operative care, anesthesia, wound closure, and use of antibiotics. Although there is a particular focus on salmonid smolts given the large body of literature available on that group, other life-stages and species of fish are discussed where there is sufficient knowledge. Additional papers explore the role of the veterinarian in fish surgeries, the need for minimal standards in the training of fish surgeons, providing a call for more complete and transparent procedures, and identifying trends in procedures and research needs. Collectively, this body of knowledge should help to improve data quality (including comparability and repeatability), enhance management and conservation strategies, and maintain the welfare status of tagged fish.     

Surgical implantation of radio tags in three eel species (Anguilla spp.) in South Africa

Studies have reported poor survival of surgically tagged freshwater fishes in warm African waters. This study aimed to assess the applicability of using radio telemetry (and surgical implantation of tags) for Anguilla spp. Nineteen yellow eels (Anguilla bengalensis, A. marmorata and A. mossambica) were surgically implanted with radio tags between October 2018 and January 2019 in the Thukela River, South Africa. Most eels were alive 6 months after tagging, and recaptured eels displayed advanced or complete healing at the incision site. Therefore, this method appears suitable for African freshwater eels.     

Advancing the surgical implantation of electronic tags in fish: a gap analysis and research agenda based on a review of trends in intracoelomic tagging effects studies

Early approaches to surgical implantation of electronic tags in fish were often through trial and error, however, in recent years there has been an interest in using scientific research to identify techniques and procedures that improve the outcome of surgical procedures and determine the effects of tagging on individuals. Here we summarize the trends in 108 peer-reviewed electronic tagging effect studies focused on intracoleomic implantation to determine opportunities for future research. To date, almost all of the studies have been conducted in freshwater, typically in laboratory environments, and have focused on biotelemetry devices. The majority of studies have focused on salmonids, cyprinids, ictalurids and centrarchids, with a regional bias towards North America, Europe and Australia. Most studies have focused on determining whether there is a negative effect of tagging relative to control fish, with proportionally fewer that have contrasted different aspects of the surgical procedure (e.g., methods of sterilization, incision location, wound closure material) that could advance the discipline. Many of these studies included routine endpoints such as mortality, growth, healing and tag retention, with fewer addressing sublethal measures such as swimming ability, predator avoidance, physiological costs, or fitness. Continued research is needed to further elevate the practice of electronic tag implantation in fish in order to ensure that the data generated are relevant to untagged conspecifics (i.e., no long-term behavioural or physiological consequences) and the surgical procedure does not impair the health and welfare status of the tagged fish. To that end, we advocate for (1) rigorous controlled manipulations based on statistical designs that have adequate power, account for inter-individual variation, and include controls and shams, (2) studies that transcend the laboratory and the field with more studies in marine waters, (3) incorporation of knowledge and techniques emerging from the medical and veterinary disciplines, (4) addressing all components of the surgical event, (5) comparative studies that evaluate the same surgical techniques on multiple species and in different environments, (6) consideration of how biotic factors (e.g., sex, age, size) influence tagging outcomes, and (7) studies that cover a range of endpoints over ecologically relevant time periods.     

Diving behaviour of jellyfish equipped with electronic tags

Jellyfish are one of the most abundant and conspicuous membersof our coastal marine fauna and are now known to play majortrophic roles in marine systems. However, little is known aboutthe movements and behaviour of individuals. We equipped individualcompass jellyfish (Chrysaora hysoscella) (n = 15) off the Dinglecoast, Ireland, with miniature time-depth recorders to log theirdepth over periods of a few hours. Vertical movements were extensive,with all jellyfish changing their depth during tracking. A rangeof vertical movements were seen including initial diving fromthe surface down to a maximum of 29.6 m after device attachment,some jellyfish remaining near the bottom, some moving up anddown in mid-water and some moving back near the surface. Theseresults show that jellyfish actively reposition themselves inthe water column over small time-scales and open the way formore extensive studies equipping jellyfish with electronic tags.     

Physiological responses of Pacific halibut, <Emphasis Type="Italic">Hippoglossus stenolepis</Emphasis>, to intracoelomic implantation of electronic archival tags,with a review of tag implantation techniques employed in flatfishes

Management of Pacific halibut (Hippoglossus stenolepis), a long-lived flatfish, is complicated by possible ontogenic and sex-specific variation in migration. Archival tags promise the ability to help uncover long-term movement patterns at the individual level, if the tags can be retained and recovered from healthy fish. We examined fifteen individuals (69–90 cm fork length) for long-term physiological response to intracoelomic implantation of three types of archival tags: fully internal, internal with right angle protruding light stalk, and internal with straight protruding light stalk. Tags represented 0.05–0.16% of initial fish weights. Fish were reared at 10.8 ± 1.1°C for 59 weeks post-surgery. One fish died after 39 weeks from thermal stress unrelated to the surgical procedure. Temporal variation in behavior of tagged fish was indistinguishable from that of controls (n = 15 tagged, 5 controls). Treatment and control-group fish grew at similar rates. No tag expulsion or physiological response was evident in the individual that died at 39 weeks, but nine of eleven individuals dissected at the end of 59 weeks had developed internal responses. These responses ranged from deposition of fibrous protein and/or calcitic material on tag surfaces to partial or full tag encapsulation in either the visceral peritoneal layer (fully-internal tags) or the intestinal mesenteries (stalk-bearing tags). The responses were within the range reported for other pleuronectids implanted with tags of similar configuration and may have implications for design and interpretation of long-term tagging studies. Encapsulation may reduce the probability of tag recoveries even in the absence of tag expulsion, especially in species eviscerated at sea.     

Electronic tags reveal behaviour of captured and discarded fish

Internally implanted data storage tags (DST) recording hydrostatic pressure (depth) and temperature in Atlantic cod Gadus morhua were used to track the behaviour of nine individuals during fishing operations. Gadus morhua showed identifiable and characteristic behavioural responses to different capture processes. In a unique observation, one fish survived being discarded and upon return to the seabed exhibited a cessation of activity comparable to observations on stressed fish under laboratory conditions.     

Cytogenetic techniques in fish genetics

A modification of a previously described method of fish blood lymphocyte culture is described and discussed with the results of various chromosome banding protocols as applied to fish chromosomes.     

Surgical skin-marking techniques

Surgical skin-marking inks and dyes are in everyday use for designing and planning incisions in plastic and reconstructive surgery. We have traced the historical development of surgical skin-marking techniques from ancient times to the present. The biochemical characteristics of the commonly used marking agents are discussed. A three-part experiment utilizing a pig model was carried out to test the tissue inflammatory response to the various dyes and inks when used intradermally as tattoos, the persistence of such tattoos, and the ease of skin erasure for each of eight stains. Methylene blue and gentian violet are recommended as the best all-purpose marking agents. The use of proprietary inks is discouraged.     

The use of archived tags in retrospective genetic analysis of fish

Collections of historical tissue samples from fish (e.g. scales and otoliths) stored in museums and fisheries institutions are precious sources of DNA for conducting retrospective genetic analysis. However, in some cases, only external tags used for documentation of spatial dynamics of fish populations have been preserved. Here, we test the usefulness of fish tags as a source of DNA for genetic analysis. We extract DNA from historical tags from cod collected in Greenlandic waters between 1950 and 1968. We show that the quantity and quality of DNA recovered from tags is comparable to DNA from archived otoliths from the same individuals. Surprisingly, levels of cross‐contamination do not seem to be significantly higher in DNA from external (tag) than internal (otolith) sources. Our study therefore demonstrates that historical tags can be a highly valuable source of DNA for retrospective genetic analysis of fish.     

Parasites as biological tags for stock discrimination in marine fish from South American Atlantic waters

The use of parasites as biological tags in population studies of marine fish in the south-western Atlantic has proved to be a successful tool for discriminating stocks for all species to which it has been applied, namely: Scomber japonicus, Engraulis anchoita, Merluccius hubbsi and Cynoscion guatucupa, the latter studied on a broader geographic scale, including samples from Uruguayan and Brazilian waters. The distribution patterns of marine parasites are determined mainly by temperature-salinity profiles and by their association with specific masses of water. Analyses of distribution patterns of some parasite species in relation to gradients in environmental (oceanographic) conditions showed that latitudinal gradients in parasite distribution are common in the study area, and are probably directly related to water temperature. Indeed, temperature, which is a good predictor of latitudinal gradients of richness and diversity of species, shows a latitudinal pattern in south-western Atlantic coasts, decreasing southwards, due to the influence of subtropical and subantarctic marine currents flowing along the edge of the continental slope. This pattern also determines the distribution of zooplankton, with a characteristic specific composition in different water masses. The gradient in the distribution of parasites determines differential compositions of their communities at different latitudes, which makes possible the identification of different stocks of their fish hosts. Other features of the host-parasite systems contributing to the success of the parasitological method are: (1) parasites identified as good biological tags (i.e. anisakids) are widely distributed in the local fauna; (2) many of these species show low specificity and use paratenic hosts; and (3) the structure of parasite communities are, to a certain degree, predictable in time and space.     

Parasites as biological tags in population studies of demersal and pelagic fish species

Among the different techniques applied in a holistic approach for fish stock identification, the use of parasites as "biological tags" is becoming increasingly important. In this presentation, our recent studies on the use of some parasite species, identified by genetic markers, and the parasite/fauna composition, in stock identification of demersal (Merluccius merluccius), small pelagic (Trachurus trachurus), and large pelagic fish species (Xiphias gladius) are reviewed. Different species of Anisakis and Hysterothylacium were genetically identified by the application of genetic (allozyme) markers. Statistically significant differences in the spatial distribution of distinct species of Anisakis were found in the fish considered. As to the species of Hysterothylacium genetically detected, different relative proportions were detected in several Mediterranean and Atlantic samples of swordfish (X. gladius). This study demonstrates the potential value of these anisakid nematodes, at both larval and adult stages, as "biological tags" for these fish species in European waters.     

转基因鱼的研究进展与商业化前景

转基因技术为鱼类育种开辟了新的途径。目前已培育出转生长激素基因鲤、鲑和罗非鱼,转荧光蛋白基因斑马鱼与唐鱼等可稳定遗传的转基因鱼品系,其中快长转生长激素基因鱼的获得对于提高水产养殖的产量与养殖效益具有十分重要的意义。文章简要综述了转基因鱼应用研究的成就、相关技术及生态安全方面的研究进展。显微注射仍是目前基因转植的常用方法,应用转座酶或巨核酸酶介导的转基因新技术可提高基因转植效率与整合率。转基因元件的选择应尽量考虑"全鱼"基因或"自源"基因,以减少转基因鱼食用安全方面的顾虑同时也有利于转植基因的表达与生理功效的发挥。生态安全是转基因鱼商用化面临的最大问题。虽然有研究显示转基因鱼与传统的选育鱼类相比适合度较差,但由于环境与基因型间的相互作用,根据实验室获得的转基因鱼对生态影响的结果,难以预测转基因鱼一旦逃逸会对自然水生态环境产生怎样的影响。因此应建立高度自然化的环境以获得可靠的数据客观评价生态风险,有效的物理拦截、不育化处理等生物学控制策略仍是保证转基因鱼安全应用的关键措施。     

Using parasites as biological tags of fish populations: A dynamical model

A simple model of macroparasitic infections has been used to evaluate the potential use of parasites as biological tags of fish populations. In the model, the parasite-host interaction is regulated by a birth-death process, and parasites can only be acquired by the non-specific migratory host population in a particular area of the space domain. In this case, we show that parasites can be succesfully used for stocks identification and to describe the migratory routes taken by some marine fish species.