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![Primary cell culture media induces darting behavior in Creek Chub. Juvenile Creek Chub [ 5 _ T D $ D I F F ] were observed in the absence of treatment, in the presence of a control solution (distilled water), and in the presence of 1.5 mL media from cultured epithelial cells (after 24 h). Observation time was 180 s. Seconds of darting behavior was averaged for 9 trials; means were compared using a t-test. *, significantly different from control, p < 0.05.](publication/321008199/figure/fig3/AS:568008791818242@1512435257250/Primary-cell-culture-media-induces-darting-behavior-in-Creek-Chub-Juvenile-Creek-Chub.png)
Primary cell culture media induces darting behavior in Creek Chub. Juvenile Creek Chub [ 5 _ T D $ D I F F ] were observed in the absence of treatment, in the presence of a control solution (distilled water), and in the presence of 1.5 mL media from cultured epithelial cells (after 24 h). Observation time was 180 s. Seconds of darting behavior was averaged for 9 trials; means were compared using a t-test. *, significantly different from control, p < 0.05.
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In various species of fishes, the importance of visual cues in the determination of environmental threat and subsequent predator avoidance is clear. Chemical cues also play an essential role facilitating predator avoidance. Among fish in the superorder Ostariophysi, club cells in the epidermis produce an alarm substance. Damage to the skin during a...
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... for 3 min, in the presence of a control solution (distilled water) for 3 min, and finally, in the presence of 1.5 mL media, which was taken from epithelial cells after 24 h of culture ( Darting behavior in fish was significantly increased following addition of cell culture media as compared to darting behavior after addition of distilled water (Fig. 3). Responses were acute, beginning immediately after addition of the media to the tank and lasting approximately 40 ...
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... Daltons). Outros estudos sugerem a presença do sulfato de condroitina como um dos componentes das substâncias de alarme (FARNSLEY et al., 2016;BROWN et al., 2000;HINTZ et al., 2017). ...
Sinais de alarme são liberados no ambiente pelas células epidérmicas de comunidades aquáticas para indicar situações de perigo. O sulfato de condroitina é indicado como um dos compostos ativos das substâncias de alarme, as quais podem induzir, nos espécimes receptores estímulos químicos, alterações comportamentais de evasão. A potencialidade da condroitina para o desenvolvimento de uma barreira comportamental para dissuasão da ictiofauna foi avaliada previamente em ensaios laboratorias (1, 2 e 5 µg L-1) e posteriormente no tubo de sucção (6 µg L-1) de turbinas do tipo buldo na Usina Hidrelétrica, no rio Madeira – Rondônia. Nos ensaios em condições controladas com as espécies Matrinxã e Tambaqui, a natação rápida e o agrupamento foram as reações de alarme mais frequentes em ambas as espécies, sendo observado maior número de episódios em concentrações maiores. No tubo de sucção, a injeção da solução concentrada de condroitina causou uma diminuição (55%) na movimentação de peixes confinados, o que indica a dissuasão da ictifauna. Os ensaios evidenciaram a potencialidade do uso sustentável do sulfato de condroitina na proteção da ictiofauna em empreendimentos hidrelétricos.
... The fin cell line is very versatile because it has a high reproductive capacity and the success rate of cell migration is high [10,102]. At the same time, many fish cells are derived from fin tissue, such as primary cell cultures derived from the fin of the endangered Yangtze sturgeon (Acipenser dabryanus) [103]. Fish polyploid cells can be inferred by the karyotype of their fin cells. ...
Fish primary cell culture has emerged as a valuable tool for investigating the physiological roles and responses of various cell types found in fish species. This review aims to provide an overview of the advancements and applications of fish primary cell culture techniques, focusing on the profiling of physiological roles exhibited by fish cells in vitro. Fish primary cell culture involves the isolation and cultivation of cells directly derived from fish tissues, maintaining their functional characteristics and enabling researchers to study their behavior and responses under controlled conditions. Over the years, significant progress has been made in optimizing the culture conditions, establishing standardized protocols, and improving the characterization techniques for fish primary cell cultures. The review highlights the diverse cell types that have been successfully cultured from different fish species, including gonad cells, pituitary cells, muscle cells, hepatocytes, kidney and immune cells, adipocyte cells and myeloid cells, brain cells, primary fin cells, gill cells, and other cells. Each cell type exhibits distinct physiological functions, contributing to vital processes such as metabolism, tissue regeneration, immune response, and toxin metabolism. Furthermore, this paper explores the pivotal role of fish primary cell culture in elucidating the mechanisms underlying various physiological processes. Researchers have utilized fish primary cell cultures to study the effects of environmental factors, toxins, pathogens, and pharmaceutical compounds on cellular functions, providing valuable insights into fish health, disease pathogenesis, and drug development. The paper also discusses the application of fish primary cell cultures in aquaculture research, particularly in investigating fish growth, nutrition, reproduction, and stress responses. By mimicking the in vivo conditions in vitro, primary cell culture has proven instrumental in identifying key factors influencing fish health and performance, thereby contributing to the development of sustainable aquaculture practices.
... Candidate compounds that may serve as one of the active ingredients of epidermal alarm cue include hypoxanthine 3N oxide [55][56][57], chondroitin sulfate [58,59], or other unidentified polypeptides [60,61]. Further, there is evidence that the source may be, in part, from epidermal club cells [53,54,62] (but see [63]) or resident surface bacteria [64]. Pupfish are in the superorder Acanthopterygii, possess some epidermal club cells [54], but the biochemistry of skin extract and alarm cue is unstudied in this group. ...
We tested whether Shoshone pupfish Cyprinodon nevadensis shoshone and Amargosa River pupfish C. n. amargosae respond behaviourally to conspecific chemical alarm cues released when epidermal tissue is damaged by a predator. We found that both subspecies reduced activity and vertical position in the water column in response to alarm cues. We then tested if pupfish can use alarm cue to acquire recognition of a novel predator. We trained pupfish with (1) water + odour of largemouth bass fed a diet of earthworms, (2) alarm cues from skin extract (epidermal alarm cues) + odour of bass fed a diet of earthworms, or (3) water + odour of bass fed a diet of pupfish (dietary alarm cues). Pupfish responded to epidermal alarm cues but not to dietary alarm cues. Pupfish were retested with the odour of bass that were fed an earthworm diet. Pupfish that had previously received epidermal alarm cues reduced vertical position and activity relative to the other two treatments. This is the first demonstration of acquired recognition of a novel predator by a pupfish, the first report of partial predator naiveté, and opens the possibility of predator-recognition training as a tool for management and conservation of endangered desert fishes.
... Os dados também indicam que as substâncias de alarme incluem oligossacarídeos de condroitina, com um tamanho mínimo de um tetrassacarídeo (~ 1.000 Daltons). Outros estudos também sugerem a condroitina e outros componentes adicionais, possivelmente compostos com óxido de nitrogênio (BROWN et al., 2000;HINTZ et al., 2017). Chia et al. (2019) ainda afirmam que pode haver um componente bacteriano nas substâncias de alarme em peixes. ...
In hydroelectric plants, there are, on a regular or untimely basis, shutdowns of the generating units in order to carry out some maneuvers for tests and mechanical maintenance. The low operating flow increases the probability of accumulation of ichthyofauna in the draft tube. As a result, the variables that determine the quality of water can change, which requires a broad and coordinated human effort to rescue trapped fish. In addition to the risks related to work safety, there are large economic liabilities as a result of the downtime of the generating units. To minimize this problem, it is necessary to improve techniques for repelling fish from risk areas. The detection of chemical substances in water is one of the most efficient methods of communication between fish. Thus, this study presents a brief review of alarm substances, which are released by the fish epidermis as a sign of defensive response to a dangerous situation and are promising for use as a chemical barrier in the hydroelectric sector.
... While low molecular weight (330-550 Da) substances did not induce an alarm behavior, substances of high molecular weight (>1500 Da) did [76]. In a non-invasive procedure, cell-free media obtained from primary culture of skin cells scraped from fish skin induced darting behavior in creek chub, indicating that injury or blood components are perhaps not required for this behavior [77]. Skin extract derived from young fathead minnows before the skin contains ECCs also induces alarm reactions [21]. ...
Epidermal club cells (ECCs), along with mucus cells, are present in the skin of many fishes, particularly in the well-studied Ostariophysan family Cyprinidae. Most ECC-associated literature has focused on the potential role of ECCs as a component of chemical alarm cues released passively when a predator damages the skin of its prey, alerting nearby prey to the presence of an active predator. Because this warning system is maintained by receiver-side selection (senders are eaten), there is want of a mechanism to confer fitness benefits to the individual that invests in ECCs to explain their evolutionary origin and maintenance in this speciose group of fishes. In an attempt to understand the fitness benefits that accrue from investment in ECCs, we reviewed the phylogenetic distribution of ECCs and their histochemical properties. ECCs are found in various forms in all teleost superorders and in the chondrostei inferring either early or multiple independent origins over evolutionary time. We noted that ECCs respond to several environmental stressors/immunomodulators including parasites and pathogens, are suppressed by immunomodulators such as testosterone and cortisol, and their density covaries with food ration, demonstrating a dynamic metabolic cost to maintaining these cells. ECC density varies widely among and within fish populations, suggesting that ECCs may be a convenient tool with which to assay ecoimmunological tradeoffs between immune stress and foraging activity, reproductive state, and predator–prey interactions. Here, we review the case for ECC immune function, immune functions in fishes generally, and encourage future work describing the precise role of ECCs in the immune system and life history evolution in fishes.
... This problem was resolved when these cells were found to function as part of the immune system; that is, they perform a function that accrues fitness benefits to the individual that possesses them (Chivers et al., 2007). Epidermal club cells are best understood as being primarily maintained by their immune function in response to epidermal damage, but because they are located in the skin they became secondarily incorporated into an alarm cue system through receiver-side selection for detection and response to components of skin released by predator attack (Hintz, Courtney, Rachel, David, & Winnifred, 2017). Thus, until Bairos- ...
In Focus: Bairos‐Novak, K.R., Ferrari, M.C.O., & Chivers, D.P. (2019). A novel alarm signal in aquatic prey: Familiar minnows coordinate group defences against predators through chemical disturbance cues. Journal of Animal Ecology, 88, 1281–1290, https://doi.org/10.1111/1365-2656.12986. Chemicals released during predation have long been studied as cues to nearby prey that predators are active in the vicinity. Until now, these chemicals have been labelled as cues because there was no compelling evidence for the necessary components of a communication system, namely (a) voluntary control of release of information, (b) capacity for graded responses and (c) the presence of specialized structures for the production and release of the signal. New findings by Bairos‐Novak, Ferrari, and Chivers (2019) show that fathead minnows alter the potency of disturbance “cues” when in the presence of other fathead minnows compared to when they are alone and produce either more or different disturbance “cues” when in the presence of familiar conspecifics compared to when they are in the presence of unfamiliar conspecifics. The behavioural response to these cues is shoaling, which would confer fitness benefits to the sender, thereby qualifying as a signal rather than a cue. This is a significant advancement in the field of chemical ecology of aquatic organisms because disturbance “cues” by fathead minnows bear two of the three hallmarks of an incipient disturbance “signal”.
Alike other flat fish, marine turbot has the particularity that changes from larvae with bilateral symmetry to adult with asymmetry, in terms of the position of the eyes. As expected, the skin configuration of this species is also affected by the development and transformation suffered by fish during metamorphosis. In this context, changes in the epidermis of marine turbot were studied using conventional staining and histochemical techniques using six lectins (UEA‐I, PNA, RCA‐I, WGA, Con A and SBA). During development from larvae to juvenile (3–300 days post‐hatching), the epidermis increased in both thickness and the number of cell layers. In fact, the simple cuboidal epithelium observed in larvae at day 3 already became stratified at days 10–12, which sequentially increase in thickness with fish development. Turbot epidermis is composed basically of four cell types: epithelial and mucous or secretory cells that are present through the development, and pigmented cells and a type that the authors described as club‐like cells that appear during and post‐metamorphosis. The Alcian blue‐periodic acid Schiff (AB‐PAS) histochemical method revealed the presence of neutral glycoconjugates in mucous and club‐like cells at post‐metamorphic stages of fish. Accordingly, lectin analysis showed mucous cells containing glycoproteins rich in fucose (UEA‐I labelling) and glycoconjugates rich in the sequence galactose‐N‐acetyl galactosamine (PNA and RCA‐I labelling) when this cell type appears. Interestingly, melanophores were observed in the dorsal epidermis of post‐metamorphic juveniles. This type of cell contains a black‐to‐brown pigment that provides the skin the typical colour of this fish species. Changes in mucous coat composition were observed during fish development, which was attributed to different roles of the glycoconjugates.
