ZebrafishPub Date : 2021-12-01DOI: 10.1089/zeb.2021.0040
Wren Wakeman, Alyssa Long, Anne M Estes, Anna K S Jozwick
{"title":"Zebrafish, <i>Danio rerio</i>, Skin Mucus Harbors a Distinct Bacterial Community Dominated by Actinobacteria.","authors":"Wren Wakeman, Alyssa Long, Anne M Estes, Anna K S Jozwick","doi":"10.1089/zeb.2021.0040","DOIUrl":"https://doi.org/10.1089/zeb.2021.0040","url":null,"abstract":"<p><p>The skin mucus of teleost fish harbors a complex microbial community that is continually interacting with the aquatic environment. Despite zebrafish, <i>Danio rerio</i>, serving as a model organism in a myriad of research fields, very little is known about the composition and role of the skin mucus microbiome. The purpose of this study was to determine a simple sampling method for the skin mucus microbiome, identify prominent bacterial members, and compare its composition to the microbial community of the surrounding environment. Next-generation sequencing of the V3-V4 region of the 16S rRNA gene was performed on skin mucus and filtered tank water samples. Results show that prominent bacterial members of the skin mucus in zebrafish include Actinobacteria (<i>Mycobacteriaceae</i>) and Gammaproteobacteria (<i>Aeromonadaceae</i>), followed by Alphaproteobacteria and Betaproteobacteria. The tank water contained much higher bacterial diversity and was clearly different from the skin mucus microbiome, despite continuous interaction. This study identifies a straightforward sampling method for the zebrafish skin mucus microbiome, enabling hypothesis generation on the role of ectosymbionts on host and microbiome health.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39747988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-12-01DOI: 10.1089/zeb.2021.0062
Yijie Geng, Randall T Peterson
{"title":"Rapid Mounting of Zebrafish Larvae for Brain Imaging.","authors":"Yijie Geng, Randall T Peterson","doi":"10.1089/zeb.2021.0062","DOIUrl":"10.1089/zeb.2021.0062","url":null,"abstract":"<p><p>Brain imaging requires mounting of zebrafish larvae in a vertical position, but anesthetized or fixed larvae tend to fall on their sides without external support. Current solution is to manually hold individual larva until liquid agarose solidifies, which is time consuming, labor intensive, and unfriendly to beginners. We developed a method to form larva-shaped slots in agarose gel using a computer numerical controlled manufactured mold. Each slot nearly perfectly fits a larva in its upright position, and larvae can be easily mounted by inserting into the slots. On average, each larva can be mounted in <1 min using this method.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716516/pdf/zeb.2021.0062.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39747987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-12-01DOI: 10.1089/zeb.2021.0071
Alex Dornburg, Tatsuya Ota, Michael F Criscitiello, Irene Salinas, J Oriol Sunyer, Susana Magadán, Pierre Boudinot, Zhen Xu, Martin F Flajnik, Amy Singer, Francisco Gambón-Deza, John D Hansen, Jeffrey A Yoder
{"title":"From IgZ to IgT: A Call for a Common Nomenclature for Immunoglobulin Heavy Chain Genes of Ray-Finned Fish.","authors":"Alex Dornburg, Tatsuya Ota, Michael F Criscitiello, Irene Salinas, J Oriol Sunyer, Susana Magadán, Pierre Boudinot, Zhen Xu, Martin F Flajnik, Amy Singer, Francisco Gambón-Deza, John D Hansen, Jeffrey A Yoder","doi":"10.1089/zeb.2021.0071","DOIUrl":"https://doi.org/10.1089/zeb.2021.0071","url":null,"abstract":"Ray-finned fishes comprise more than half the *60,000 known vertebrate species, and are pivotal to the functionality of aquatic ecosystems and success of global multibillion dollar industries. Understanding ray-finned fish immune systems is essential to predicting how species will respond to known or emergent pathogens as well as to the development of effective vaccines for aquaculture. However, the diversity of species, including in aquaculture, necessitates that immunology and translational medicine research groups investigating the immune system in one or a number of species employ a common language for describing homologous immune components. Unfortunately for Immunoglobulin (Ig) genes that encode antibodies, this has not been the case. Ig genes are restricted to jawed vertebrates (gnathostomes) with all lineages encoding common heavy chains IgM and IgD (aka IgW in cartilaginous fish, lungfish, and coelacanths). Before 2005, it was believed that bony fish encoded only IgM and IgD. In 2005, Hansen et al. described a new Ig heavy chain encoded within the rainbow trout heavy chain locus and named it IgT for ‘‘teleost.’’ However, during the","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716470/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39747986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-12-01Epub Date: 2021-09-07DOI: 10.1089/zeb.2021.0031
Gokul Kesavan, Anja Machate, Michael Brand
{"title":"CRISPR/Cas9-Based Split Fluorescent Protein Tagging.","authors":"Gokul Kesavan, Anja Machate, Michael Brand","doi":"10.1089/zeb.2021.0031","DOIUrl":"https://doi.org/10.1089/zeb.2021.0031","url":null,"abstract":"<p><p>Genetically encoded fluorescent tags such as green fluorescent protein fused to protein have revolutionized cell biology as they permit high-resolution protein imaging in live systems. Split fluorescent proteins, with a small fragment of 16 amino acids, can be inserted in the coding sequence to label proteins. We demonstrate successful integration of two bright and fast maturing split fluorescent proteins, mNeon green and sfCherry2, in zebrafish, and show that they are suitable for live imaging, including time-lapse series, and that they have a high signal-to-noise ratio. Furthermore, we show that CRISPR/Cas9 can be used to generate fluorescently tagged proteins <i>in vivo</i>.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39394960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-12-01Epub Date: 2021-09-20DOI: 10.1089/zeb.2021.0030
Toi A Collins, Shelby Cabrera, Emily Teets, Jami Shaffer, Bradley W Blaser
{"title":"An Optimized Zebrafish Nursery Feeding Regimen Improves Growth Rates and Labor Costs.","authors":"Toi A Collins, Shelby Cabrera, Emily Teets, Jami Shaffer, Bradley W Blaser","doi":"10.1089/zeb.2021.0030","DOIUrl":"https://doi.org/10.1089/zeb.2021.0030","url":null,"abstract":"<p><p>Setting nutritional standards for larval zebrafish (<i>Danio rerio</i>) that maximize growth, survival, and reproductive success is challenging. We evaluated the effects of different feeding regimens on larval zebrafish by comparing Gemma Micro 75 pelleted diet and live-type L rotifers (<i>Brachionus plicatilis</i>) in 3 feeding regimens starting at 9 days postfertilization (dpf): bolus feeding of live diet (BL), continuous feeding of live diet (CL), and pelleted diet (PD). Animals in the PD and CL groups were longer than the BL group at 4-5 weeks postfertilization. The PD group was also greater in body depth than both live diet groups. There was no significant difference in weight between the groups. There were also no significant differences in fecundity or sex ratios indicating that all feeding methods successfully promote growth of a useful breeding stock of fish. In addition, we quantified the equipment, consumable, and labor costs associated with these methods, and found that the PD regimen was superior to both live diet regimens. These data suggest that providing a high nutrient-density pelleted diet to larval and juvenile zebrafish is an effective means to increase early growth and to decrease cost and labor associated with nursery care.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716471/pdf/zeb.2021.0030.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39431591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-12-01Epub Date: 2021-10-20DOI: 10.1089/zeb.2021.0059
Robert D Morabito, Rebecca C Adikes, David Q Matus, Benjamin L Martin
{"title":"Cyclin-Dependent Kinase Sensor Transgenic Zebrafish Lines for Improved Cell Cycle State Visualization in Live Animals.","authors":"Robert D Morabito, Rebecca C Adikes, David Q Matus, Benjamin L Martin","doi":"10.1089/zeb.2021.0059","DOIUrl":"https://doi.org/10.1089/zeb.2021.0059","url":null,"abstract":"Adikes et al. 1 described a novel zebrafish cell cycle sensor that delineates all phases of the cell cycle based on levels of cyclin-dependent kinase (CDK) activity. The CDK sensor consists of a fragment of human DNA Helicase B (DHB) fused to a fluorescent protein. DHB contains a dominant nuclear localization sequence (NLS) and a nuclear export sequence (NES) flanked by CDK-specific phosphorylation sites. Cells that contain low levels of CDK activity have an exposed NLS that localizes the CDK sensor to the nucleus. As the cell cycle progresses, CDK activity increases causing phosphorylation of DHB, which occludes the NLS and allows the NES to promote export of the sensor into the cytoplasm (Fig. 1). The authors showed that quantitative analysis of this ratiometric sensor can distinguish CDK low (CDK), quiescent G0 arrested cells from CDK increasing (CDK), cycling G1 phase cells, as well as identify S, G2, and M phases of the cell cycle.","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716472/pdf/zeb.2021.0059.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39533874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-10-01Epub Date: 2021-07-26DOI: 10.1089/zeb.2021.0023
Sara Jorge, Jorge M Ferreira, I Anna S Olsson, Ana M Valentim
{"title":"Adult Zebrafish Anesthesia: A Study of Efficacy and Behavioral Recovery of Different Anesthetics.","authors":"Sara Jorge, Jorge M Ferreira, I Anna S Olsson, Ana M Valentim","doi":"10.1089/zeb.2021.0023","DOIUrl":"https://doi.org/10.1089/zeb.2021.0023","url":null,"abstract":"<p><p>The use of proper anesthesia in zebrafish research is essential to ensure fish welfare and data reliability. However, anesthesia long-term side effects remain poorly understood. The purpose of this study was to assess anesthesia quality and recovery in adult zebrafish using different anesthetic protocols and to determine possible long-term effects on the fish activity and anxiety-like behaviors after anesthesia. Mixed-sex adult AB zebrafish were randomly assigned to five different groups (Control, 175 mg/L of tricaine methanesulfonate [MS222], 45 mg/L of clove oil, 2 mg/L of etomidate, and 5 mg/L of propofol combined with 150 mg/L of lidocaine) and placed in the respective anesthetic bath. Time to lose the equilibrium, response to touch and to caudal fin pinch stimuli, and recovery after anesthesia administration were evaluated. In addition, after stopping anesthesia, respiratory rate, activity, and anxiety-like behaviors in the novel tank test were studied. Overall, all protocols proved to be adequate for zebrafish anesthesia research as they showed full recovery at 1 h, and only etomidate had minor effects on fish behavior in the novel tank, a validated test for anxiety.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39226338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-10-01Epub Date: 2021-08-10DOI: 10.1089/zeb.2021.0012
Ishani Mukherjee, Anuradha Bhat
{"title":"Temporal Patterns in Interactions Across Same- and Mixed-Sex Wild Zebrafish Dyads.","authors":"Ishani Mukherjee, Anuradha Bhat","doi":"10.1089/zeb.2021.0012","DOIUrl":"https://doi.org/10.1089/zeb.2021.0012","url":null,"abstract":"<p><p>Animals communicate with each other through a variety of behavioral interactions, many of which are often complex due to the interplay of several ecological factors. Observations on dyadic interactions can help throw light on the more complex interactions observed among group living organisms and can help in understanding mechanisms of behaviors related to mating strategies, dominance hierarchies, and decision-making. This study focused on the assessment of several generally observed interactions among dyads of different sexes (female-female, male-male, and male-female) in wild zebrafish (<i>Danio rerio)</i>. Temporal dynamics of these interactive behaviors were observed in 45 dyads across 3 time intervals of the day. We used generalized linear mixed models to investigate the effect of time, sex of dyad, and their interaction on specific behaviors. While the frequency of occurrence of some behaviors showed clear variation across time intervals of the day, these were further found to depend on the composition of the dyad. Contrary to previous reports, we found that same-sex dyads are equally aggressive and aggressive interactions did not vary temporally. Mating-associated interactions, as expected, were significantly higher in mixed-sex dyads and declined significantly from early morning to afternoon. Interestingly, we also found some mating-associated interactions in same-sex dyads. A fine line exists between social and mating-associated interactions in many organisms and so we speculate that these interactions could also be social interactions and not mating-related behavior. Our findings shed light on complex interactive behaviors among zebrafish, that are likely to be affected by time as well as sex composition of interacting individuals and thus has important implications for groups varying in sex ratios in the wild.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39300783","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Control of Developmental Speed in Zebrafish Embryos Using Different Incubation Temperatures.","authors":"Hirotaro Urushibata, Kazuaki Sasaki, Eisuke Takahashi, Toshikatsu Hanada, Takafumi Fujimoto, Katsutoshi Arai, Etsuro Yamaha","doi":"10.1089/zeb.2021.0022","DOIUrl":"https://doi.org/10.1089/zeb.2021.0022","url":null,"abstract":"<p><p>The zebrafish is a valuable model organism that is widely used in studies of vertebrate development. In the laboratory, zebrafish embryonic development is normally carried out at 28.5°C. In this study, we sought to determine whether it was possible to modify the speed of embryonic development through the use of short- and long-term variations in incubation temperature. After incubation at 20°C-32°C, most early-stage embryos survived to the epiboly stage, whereas more than half of the embryos died at <20°C or >32°C. The rate of development differed between embryos incubated at the lowest (18°C) and highest (34°C) temperatures: a difference of 60 min was observed at the 2-cell stage and 290 min at the 1k-cell stage. When blastulae that had developed at 28°C were transferred to a temperature lower than 18°C for one or more hours, they developed normally after being returned to the original 28°C. Analyses using green fluorescent protein-<i>buckyball</i> mRNA and <i>in situ</i> hybridization against <i>vasa</i> mRNA showed that primordial germ cells increase under low-temperature culture; this response may be of use for studies involving heterochronic germ cell transplantation. Our study shows that embryonic developmental speed can be slowed, which will be of value for performing time-consuming, complicated, and delicate microsurgical operations.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39391497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ZebrafishPub Date : 2021-10-01Epub Date: 2021-08-26DOI: 10.1089/zeb.2021.0018
Marica Andersson, Petronella Kettunen
{"title":"Effects of Holding Density on the Welfare of Zebrafish: A Systematic Review.","authors":"Marica Andersson, Petronella Kettunen","doi":"10.1089/zeb.2021.0018","DOIUrl":"https://doi.org/10.1089/zeb.2021.0018","url":null,"abstract":"<p><p>The zebrafish is becoming an increasingly popular research animal around the world. Its welfare is affected by an array of environmental factors, such as food access and water quality. Holding density is an important welfare aspect, not least due to its interaction with other housing conditions. Despite the extensive use of zebrafish in research, little is known of how densities affect its welfare. In this systematic review, we have performed a large literature search, compiled, and evaluated all publications regarding zebrafish holding density. We have analyzed how density effects growth, reproduction, and stress response, including behavior, water quality, and pathogenic outbreaks in young and adult fish. Our review shows that the holding densities tested vary largely depending on the research focus, for example, body growth or behavior. In fact, research indicates that future recommendations on holding density could depend on which welfare aspects are considered. Overall, there is a need for more studies investigating the interactive effects of density on welfare indicators, such as reproduction coupled with stress response. We stress the necessity of including holding density in universal housing guidelines and reporting information on holding conditions of larvae and adults when publishing zebrafish work.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2021-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39357322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}