From noopur.mandrekar from gmail.com Wed Jul 2 05:10:00 2008 From: noopur.mandrekar from gmail.com (Noopur) Date: Wed Jul 2 09:50:36 2008 Subject: [Zbrafish] Cyp activity Message-ID: <5a0f93da-f00a-4be7-8d87-56be80187ae5@w8g2000prd.googlegroups.com> Dear All, I am looking out for a method to evaluate the Cyp3A4 functional (enzyme) activity using Zebrafish larvae. Can anyone guide me in this matter? Thanking you Regards Noopur M From sspinette from ric.edu Wed Jul 2 07:29:59 2008 From: sspinette from ric.edu (sarah) Date: Wed Jul 2 09:50:56 2008 Subject: [Zbrafish] Re: Microsporidiosis References: Message-ID: <825b7f40-a490-48c9-9109-8bc26913b6c3@79g2000hsk.googlegroups.com> On Jun 30, 1:38?pm, Katy wrote: > On Jun 25, 10:32 am, sarah wrote: > > > I suspect that we may have some fish with Microsporidiosis infection.>From reading that I have done it seems that I should euthenize ALL of > > > the fish in the AHAB benchtop system and empty all water and clean all > > tanks and resevoir. But I have also read that this parasite may not be > > killed by common disinfection agents. How can I completely rid the > > system of this agent? can things be taken apart and autoclaved? Has > > anyone else encountered this and what did you do? ?PLEASE HELP!!! > > Hi Sarah, > First, I would confirm that you do in fact have microsporidiosis > (Pseudoloma neurophilia) in your zebrafish. ?Almost any pathogen can > result in a skinny fish and there is no pre-mortem diagnostic for > Pseudoloma in zebrafish. ?You can diagnose microsporidiosis in H&E > stained histological sections, wet mounts of the spinal cord and > brain, or by PCR on spinal cord and brain tissue. > > Bleach concentrations used to sanitize eggs are not effective against > Pseudoloma spores. ?However, high concentrations of pH adjusted bleach > will kill the spores. ?See Ferguson et al. (2007), Dis Aquat Org, Vol. > 76: 205-214. ?Bleach is toxic to zebrafish, causing acute gill > necrosis and asphyxiation. ?Therefore, it's important to neutralize > with sodium thiosulfate and rinse well. ? Having said that, the most > likely, and proven, mode of transmission is by cannibalism of dead, > infected fish. ?Therefore, removing dead, old, and sick fish from your > population is the most effective way to break the cycle of > transmission. ?I am not aware of Pseudoloma being detected in biofilms > and have not personally been able to detect it in water or debris > samples from tanks with infected fish. > > UV is also an effective means of killing Pseudoloma spores. ?Have you > checked the efficacy of your UV bulbs? ?I suspect that dead fish can > get pulverized in the drains and filtration system, liberating > infective spores. ?However, UV will kill the spores and prevent > circulation of Pseudoloma throughout your system. > > Katy Murray DVM, Ph.D. > Zebrafish International Resource Center Thank you very much. We do have UV but we are actually planning to replace the bulb this week for that reason. We have been removing symptomatic fish at the first detectable signs, but still we are finding new sick fish each week. Yesterday, I was doing the bi-weekly scouring of the resevoir and found a gritty fine sand-like substance inside the bottom! My student claims that it was not there 2 weeks ago. I have no idea where it could have come from aside from the water we use for daily water exchanges (we used to use conditioned tap water and now use conditioned filtered water). It was the color and feel of the Siprax beads ...do these ever start to break down? From schneiderac from gmail.com Wed Jul 2 13:55:42 2008 From: schneiderac from gmail.com (Ana Schneider) Date: Wed Jul 2 14:07:00 2008 Subject: [Zbrafish] Cyp activity In-Reply-To: <5a0f93da-f00a-4be7-8d87-56be80187ae5@w8g2000prd.googlegroups.com> References: <5a0f93da-f00a-4be7-8d87-56be80187ae5@w8g2000prd.googlegroups.com> Message-ID: <3c92106a0807021155r1a990dbfxd258fdf0787841c9@mail.gmail.com> Dear all! I am looking for a method to evaluate CYP 450 in liver of Zebrafish. Does anyone know any work about this? Thanks, Kind regards Ana Schneider** Experimental Laboratory of Hepatology and Gastroenterology - Research Center - HCPA POA/RS - BRAZIL Fone: 21018847 On Wed, Jul 2, 2008 at 7:10 AM, Noopur wrote: > Dear All, > > I am looking out for a method to evaluate the Cyp3A4 functional > (enzyme) activity using Zebrafish larvae. Can anyone guide me in this > matter? > > Thanking you > > Regards > Noopur M > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > -- Ana Cl?udia Schneider ~~~~~~~~~~~~~~~~~~~~~~ ?.??.???`?.?.???`?...?><)))?> . , . .???`?.. ><)))?> ><> <>< ><> <>< ><> <>< ><> <>< -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080702/60859678/attachment.html From tgenade from gmail.com Fri Jul 4 02:06:55 2008 From: tgenade from gmail.com (Tyrone) Date: Mon Jul 7 11:07:47 2008 Subject: [Zbrafish] Re: Cyp activity References: <5a0f93da-f00a-4be7-8d87-56be80187ae5@w8g2000prd.googlegroups.com> Message-ID: <694dea51-d1ba-41ac-b6b2-0b811e2c8c31@27g2000hsf.googlegroups.com> Hello, On Jul 2, 8:55?pm, "Ana Schneider" wrote: > I am looking for a method to evaluate CYP 450 in liver of Zebrafish. Does > anyone know any work about this? P450 enzymes, such as CYP3A4, were subject of my masters thesis (though I worked on steroid synthesizing P450s of the adrenal) which is accesible at http://tgenade.freeshell.org (you will have to explore a bit). My thesis includes a protocol for the isolation of microsomal P450s as well as protocol for P450 concentration estimation and microsomal P450 enzyme assays in microL quantities. You will probably need to isolate the enzyme which means you will have to liquidize large numebers of zebrafish fry. It is possible that the protocol can be downscaled but much optimization will be needed. You may want to look into dissecting out embryonic livers (good luck... you will have to find someone else to help with that, I have no idea if it is even possible) and then culturing them up, but I do not know if you will get representative data. I gather you are trying look at induction of the enzyme under physiological conditions in response to some environmental insult, suggesting that enzyme isolation may be the way to go. You may want to pick the brains of my ex-professor: Piet Swart. His email is (was?) pswart@sun.ac.za. The isolation protocol is rather generic, I think your biggest problem will be scaling the P450 concentration and enzyme assays to meet your requirments. I used about half a kg of sheep adrenals to get a couple of mg of P450. I cannot even begin to speculate on how many zebrafish fry you will need! Kind regards Tyrone From Thomas.Vihtelic from mpiresearch.com Mon Jul 7 11:17:48 2008 From: Thomas.Vihtelic from mpiresearch.com (Thomas Vihtelic) Date: Mon Jul 7 11:26:00 2008 Subject: [Zbrafish] email change Message-ID: To whom it may concern: Please change my email address (formerly "vihtelic.3@nd.edu") To thomas.vihtelic@mpiresearch.com Thank you. Thomas S. Vihtelic, DVM, PhD Study Director, Targeted Discovery Research MPI Research, Inc. 54943 North Main Street Mattawan, MI 49071-9399 Tel: (269) 668-3336, x2266; Fax: (269) 668-4151 Office: M-3125 thomas.vihtelic@mpiresearch.com This communication, including attachments, is for the exclusive use of addressee and may contain proprietary, confidential and/or privileged information. If you are not the intended recipient, any use, copying, disclosure, dissemination or distribution is strictly prohibited. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this communication and destroy all copies. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080707/a2f63a01/attachment.html From els.janssens from gmail.com Tue Jul 8 08:53:26 2008 From: els.janssens from gmail.com (Els) Date: Tue Jul 8 12:01:49 2008 Subject: [Zbrafish] albino zebrafish Message-ID: Hi, Since I'm interested in testing some eye-specific proteins in the zebrafish I'm looking for a homozygous albino zebrafish line. Does anyone know a source where I can buy them? (adults and/or embryos) Greetz, Els Els Janssens PhD-student Animal Physiology and Neurobiology Department Neural Circuit Development and Regeneration Naamsestraat 61, bus 2464 B-3000 Leuven Belgium Phone: +32 16 323987 Fax: +32 16 324262 E-mail: els.janssens@bio.kuleuven.be From els.janssens from gmail.com Tue Jul 8 08:59:49 2008 From: els.janssens from gmail.com (Els) Date: Tue Jul 8 12:02:07 2008 Subject: [Zbrafish] whole mount ISH Message-ID: Hi, Does anyone ever done a whole mount ISH on 5dpf embryos that worked? I already tried practically every protocol but none of them are working. Greetz, Els Janssens Els Janssens PhD-student Animal Physiology and Neurobiology Department Neural Circuit Development and Regeneration Naamsestraat 61, bus 2464 B-3000 Leuven Belgium Phone: +32 16 323987 Fax: +32 16 324262 E-mail: els.janssens@bio.kuleuven.be From nooria from univmail.cis.mcmaster.ca Tue Jul 8 11:58:27 2008 From: nooria from univmail.cis.mcmaster.ca (A. Noori) Date: Tue Jul 8 12:09:18 2008 Subject: [Zbrafish] yolk sac injection Message-ID: Hello everyone, our lab is working on an integrated device for injection of Zebrafish embryos. We currently lack the ability to orient the embryo, so to start off we would like to do viability studies that involve injections into the yolk sac. I was wondering what potential applications there are for injections into the yolk sac. Thanks, Arash Noori ------------------------------------------ M.A.Sc. Candidate Centre for Advanced Micro-Electro-Fluidics Mechanical Engineering McMaster University +1-905-525-9140 x21388 ------------------------------------------ From david from zebrafish.org Tue Jul 8 12:44:29 2008 From: david from zebrafish.org (David Lains) Date: Tue Jul 8 12:48:54 2008 Subject: [Zbrafish] albino zebrafish In-Reply-To: References: Message-ID: <08b001c8e122$49141650$db3c42f0$@org> Hello Els We have albino^b4 available at the Zebrafish International Resource Center. Currently we have heterozygous incrossed embryos available. I would be glad to ship to you if you can not find homozygous fish. Best Fishes David Lains <}}}>< Aquaculturist, Research Assistant Zebrafish International Resource Center 5274 University of Oregon Eugene, Or 97403 Email: david@zebrafish.org pH: (541) 346-6028 ext. 18 fax: (541) 346-6151 -----Original Message----- From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Els Sent: Tuesday, July 08, 2008 6:53 AM To: bionet-organisms-zebrafish@moderators.isc.org Subject: [Zbrafish] albino zebrafish Hi, Since I'm interested in testing some eye-specific proteins in the zebrafish I'm looking for a homozygous albino zebrafish line. Does anyone know a source where I can buy them? (adults and/or embryos) Greetz, Els Els Janssens PhD-student Animal Physiology and Neurobiology Department Neural Circuit Development and Regeneration Naamsestraat 61, bus 2464 B-3000 Leuven Belgium Phone: +32 16 323987 Fax: +32 16 324262 E-mail: els.janssens@bio.kuleuven.be _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From finchg from ohsu.edu Wed Jul 9 17:04:45 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Wed Jul 9 18:17:53 2008 Subject: [Zbrafish] Re: whole mount ISH References: Message-ID: <74326fb6-226c-47a1-8547-6e53e36476d5@y21g2000hsf.googlegroups.com> Els, When you say none of them are working do you mean no signal or embryo- wide signal? Also, are you sure your gene of interest is expressed at the 5dpf timepoint as a lack of signal may not necessarily indicate a defective protocol? Having tried several protocols for zebrafish in situ and read through nearly every one I could find on the internet, I would recommend the adapted Thisse et al. protocol available from the Moens Lab website: . This protocol has worked consistantly even at 5dpf timepoint. For 5d embryos, use of PTU from ~12 hrs. to prevent pigment formation is superior to later (potentially destructive) post-fix bleaching. In preparing the fish for hybridization, protienase K digestion time may need to be optimized due to product variability. I use Roche PCR grade ProK at 10ug/ml and would recommend a digest time of at least 30 min for 5dpf. Overdigestion is apparent when the embryos break apart, underdigestion may be less evident, but will effect staining of deep tissues. If you are concerned only with superficial structures than fine tuning this may be less relevant. I also have much more consistant results mixing my own staining buffer using Roche NBT/BCIP (as in the recommended protocol) as opposed to the Roche hybridization kit which uses "BM Purple AP substrate". If you have access to a probe that you know works, try it at the same time as any untested/troublesome probes as a positive control. This way you can quickly determine if problems are with probe preparation or with some other aspect of the chemistry. The same could be said for trying another earlier "easier" time-point at the same time. -Gabe Finch Nicolson Lab OHSU Portland, Oregon On Jul 8, 6:59 am, Els wrote: > Hi, > > Does anyone ever done a whole mount ISH on 5dpf embryos that worked? > I already tried practically every protocol but none of them are > working. > > Greetz, > > Els Janssens > > Els Janssens > PhD-student > Animal Physiology and Neurobiology Department > Neural Circuit Development and Regeneration > Naamsestraat 61, bus 2464 > B-3000 Leuven > Belgium > Phone: +32 16 323987 > Fax: +32 16 324262 > E-mail: els.janss...@bio.kuleuven.be From naruse from nibb.ac.jp Thu Jul 10 01:49:11 2008 From: naruse from nibb.ac.jp (Kiyoshi) Date: Thu Jul 10 11:39:25 2008 Subject: [Zbrafish] Renewal of Medaka fish BioResource Project (NBRP Medaka) website Message-ID: <007601c8e259$0f433a90$2dc9afb0$@ac.jp> Renewal of Medaka fish BioResource Project (NBRP Medaka) website (http://www.shigen.nig.ac.jp/medaka/). We are pleasure announcing the renewal of NBRP medaka website. New website implemens several search system including keywords and blast search to the live fish resource (656 strains), the full length cDNA (265859 entries) and BAC/fosmid end sequence (551547 entries) resources and others. You can search the BAC/Fosmid clones with the gene of interest by following the description in the laboratory manual (http://www.shigen.nig.ac.jp/medaka/strain/identify.jsp) and order these fish and clones from new website. Direct order of clones identified by DDBJ/Genbank/EMBL or DFCI Medaka Gene Index(http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=o_latip es) is welcome. You can see the manual for medaka research (medaka book), anatomical atlas of brain and vessel system (atlas), phylogenetic relationship of medaka wild population and the relates species (medaka tree) and medaka genome browser using ensembl interface (medaka map). Deposition of your medaka fish strain to NBRP medaka is welcome. Please visit our website for details (http://www.shigen.nig.ac.jp/medaka/strain/deposition.jsp). Your requests or questions are important for improving our website. NBRP medaka website is a database for yours. Email address for questions and resource order is mbrc@nibb.ac.jp Please enjoy All the best Kiyoshi ---------------------------------------- Kiyoshi NARUSE Ph.D. National Institute for Basic Biology, Laboratory of Bioresources, Nishigonaka 38, Myodaiji, Okazaki 444-8585, Aichi, Japan TEL: 0564-55-7581?TEL/FAX: 0564-55-7580 email: naruse@nibb.ac.jp ----------------------------------------------------- From zhenfh from ust.hk Thu Jul 10 06:55:09 2008 From: zhenfh from ust.hk (Zhen Fenghua) Date: Thu Jul 10 11:39:44 2008 Subject: [Zbrafish] Zebrafish: (Big problem) BAC/PAC injection into embryos Message-ID: <1609.143.89.52.224.1215690909.squirrel@sqmail.ust.hk> Dear all: I am currently doing a project to screen a transgenic fish line by injection of PAC/BAC construct into fish embryos. We found a very interesting phenomenon: the uptake of PAC DNA, or in another words, the expression of GFP signal is different in different WILD TYPE fish lines, although all the other conditions are exactly same. Does anyone have such experiences before? Anyway, the good news is that we have found a specific wild type fish line who will carry the best expression of PAC-driven GFP, but the bad news is that the next generation of this fish line get worse expression of GFP; therefore, it brings a big trouble to my experiments. So may anyone help to guess the reason or share any idea with me? My email address is zhenfh@ust.hk, and I will be very appreciated to any of your response. Thanks a lot! Best regards, zhen feng hua Tel: 852-2358-7282 Hong Kong University of Scoence and Technology From hernoc from gmail.com Tue Jul 15 12:36:33 2008 From: hernoc from gmail.com (hernoc@gmail.com) Date: Tue Jul 15 12:39:27 2008 Subject: [Zbrafish] Alternative to gene Tools Message-ID: <52ac5730-0d54-4867-937c-088740bfd5a9@m3g2000hsc.googlegroups.com> Hi Neters, does anyone know of an alternative commercial and reliable source for morpholinos? GeneTools is becoming a real pain in the *** to work with. Thanks a lot, Hernan Lopez-Schier.- From jmoulton from gene-tools.com Tue Jul 15 14:05:18 2008 From: jmoulton from gene-tools.com (Jon D.Moulton) Date: Tue Jul 15 14:22:33 2008 Subject: [Zbrafish] Alternative to gene Tools In-Reply-To: <52ac5730-0d54-4867-937c-088740bfd5a9@m3g2000hsc.googlegroups.com> References: <52ac5730-0d54-4867-937c-088740bfd5a9@m3g2000hsc.googlegroups.com> Message-ID: <487CF4EE.2020304@gene-tools.com> Dear Hernan, Regarding your newsgroup post: "Hi Neters, does anyone know of an alternative commercial and reliable source for morpholinos? GeneTools is becoming a real pain in the *** to work with. Thanks a lot, Hernan Lopez-Schier.-" Regarding your question about alternative commercial sources for Morpholino oligos, Morpholinos are patented by AVI BioPharma Inc. and are exclusively licensed to Gene Tools LLC for the research reagent market. Your institution is currently having an accounting issue with Gene Tools. We will have to let the accountants sort that out; we expect this will be resolved promptly. I apologize for any pain you have endured from this accounting issue. I sincerely hope that the quality of our oligos and customer support has met or exceeded your expectations. Let me know how I can help. Regards, - Jon Jon D. Moulton, Ph.D. Diagnostics and Special Projects GENE TOOLS, LLC jmoulton@gene-tools.com www.gene-tools.com (541) 929-7840 x1201 http://network.nature.com/group/morpholinos From stefan.weigt from merck.de Wed Jul 16 08:45:43 2008 From: stefan.weigt from merck.de (stefan1306) Date: Wed Jul 16 12:00:51 2008 Subject: [Zbrafish] Antibodies to detect necrosis Message-ID: <36c01091-ce6b-4fa3-81f2-33e0d084b63c@26g2000hsk.googlegroups.com> Dear all, at the moment I?m dealing with histology of 96 hpf embryos and I want to show necrosis in the embryos by immunohistochemistry. Do any of you know suitable antibodies (and protocols) for these purposes? Thank you very much in advance! Stefan From huangva from gmail.com Thu Jul 17 09:32:35 2008 From: huangva from gmail.com (huangva@gmail.com) Date: Thu Jul 17 12:31:31 2008 Subject: [Zbrafish] Egg water vs. embryo medium Message-ID: Hi, I got a little confused with these two types of media (from Zebrafish Book). Which one should I use for embryos at 5-8 dpf? Recently I tried to treat embryos at these stages with drugs and found their responses were very different in the two media. Besides, the survival rates were also different between the two. I'm guessing one of them is better for the embryos' health than the other? Any help on this would be appreciated. Thanks. MH From tgenade from gmail.com Fri Jul 18 08:01:19 2008 From: tgenade from gmail.com (Tyrone) Date: Fri Jul 18 12:30:04 2008 Subject: [Zbrafish] Re: Antibodies to detect necrosis References: Message-ID: Hello On Jul 16, 3:45?pm, stefan1306 wrote: > at the moment I?m dealing with histology of 96 hpf embryos and I want > to show necrosis... You may want to pose this question to the histonet group. They may be more forthcoming with useful information. You may also want to track down Maldonado et al Neurobiol. 53:21--35, 2002. In it they show neurodegeneration (via what appears to be a non-apoptotic mechanism) using cresyl violet. If you find a better means to show necrosis, please let me know! I am very far from being an expert on this topic. Kind regards From schneiderac from gmail.com Sat Jul 19 15:24:40 2008 From: schneiderac from gmail.com (Ana Schneider) Date: Mon Jul 21 12:15:21 2008 Subject: [Zbrafish] Re: Antibodies to detect necrosis In-Reply-To: References: Message-ID: <3c92106a0807191324u1d25db42gdc3e21fa0d4ca7cb@mail.gmail.com> There's an article by Amali et al (Journal of Biomedical Science (2006) 13: 225–232) where they looked for apoptosis in liver cells of embryos using TUNEL method. After 7 days they observed a diminished number of apoptotic bodies, due to the increasing number of necrotic bodies. I hope it helps you. Regards, Ana Schneider On Fri, Jul 18, 2008 at 10:01 AM, Tyrone wrote: > Hello > > On Jul 16, 3:45 pm, stefan1306 wrote: > > > at the moment I´m dealing with histology of 96 hpf embryos and I want > > to show necrosis... > > You may want to pose this question to the histonet group. They may be > more forthcoming with useful information. You may also want to track > down Maldonado et al Neurobiol. 53:21--35, 2002. In it they show > neurodegeneration > (via what appears to be a non-apoptotic mechanism) using cresyl > violet. > > If you find a better means to show necrosis, please let me know! I am > very far from being an expert on this topic. > > Kind regards > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080719/e4505b4f/attachment.html From alzaidan.abdullah from gmail.com Mon Jul 21 20:50:33 2008 From: alzaidan.abdullah from gmail.com (alzaidan.abdullah@gmail.com) Date: Tue Jul 22 12:42:01 2008 Subject: [Zbrafish] ZebraFish Skin Histology Message-ID: Hi, I’m currently studying the effect of contaminants on the skin of Zebrafish through Histology. The problem I’m facing while processing the tissue is after doing so the skin always detaches from the muscle. The fixative used is 4% NBF for 24 hrs. And I’m only removing the skin attached with muscle, also tried to cut a TS section (Dorsal to Ventral through the vertebra) but similar effect. The thickness used on microtome is 5um. Any help would be appreciated Abdullah Al-Zaidan From alzaidan.abdullah from gmail.com Wed Jul 23 00:57:07 2008 From: alzaidan.abdullah from gmail.com (alzaidan.abdullah@gmail.com) Date: Wed Jul 23 16:16:16 2008 Subject: [Zbrafish] Lysozyme Extraction from the skin mucous Message-ID: <46939644-d49b-44d1-9739-fccf006f6702@a6g2000prm.googlegroups.com> Hi, I’m currently studying the effect of contaminants on the skin of Zebrafish. I'm trying to measure the levels of Lysozyme through the skin mucous. I tried placing 10ml of 50mM NaCl in a Ziplock bag and shaking it with the fish anaesthetized inside, but after centrifuging at 1500xg 10min 4degrees coulden't see the supernatant (fish number used were 6). So any ideas? Thank you Abdullah From jason.cockington from gmail.com Tue Jul 22 20:25:34 2008 From: jason.cockington from gmail.com (Leviathan) Date: Wed Jul 23 16:16:37 2008 Subject: [Zbrafish] Physiological Saline Message-ID: Hi everyone, I am hoping that some of the physiologists out there may be able to help me here. I have just had a researcher here in adelaide approach me inquiring as to whether she could do some physiological studies on the zebrafish. Considering past researchers here have all been a twist of the molecular and developmental biology this request is all very new for me. I think that what she is proposing is grand, but she asked what the composition of the physiological saline she should use for the tissue experiments should be to mimic the internal environment of a zebrafish? i have no idea, and thought i'd ask you all, to see if anyone out there may know. if anyone can offer assistance that would be excellent :O) many thanks, jason From wclements from ucsd.edu Tue Jul 22 19:55:16 2008 From: wclements from ucsd.edu (Wilson Clements) Date: Wed Jul 23 16:16:57 2008 Subject: [Zbrafish] Re: Tol2 injections Message-ID: Dear ZF Community, We are trying to optimize our dosage for Tol2 transgenesis. I was hoping to poll people who are using this system to see roughly what amounts of transgenesis plasmid and Tol2 transposase mRNA you are injecting to obtain transgenics. For no particular reason, we started out using 50 pg of DNA and 50 pg of RNA. We find we get high levels of transgenesis and many insertions per genome. On the plus side, many insertions gives higher expression in the F1. But on the negative side, it makes it easier to see position effects, lose expression over successive generations, and makes it difficult to track specific insertions. If you have experiences you would be willing to share in optimizing your injections to whatever parameters you feel are important please send them either to the list server or to me directly at the e-mail address below. Best, Wilson ------------------------------------------------------------------------ -------------------- Wilson Clements, Ph.D. wclements@ucsd.edu Dept. of Biology Section of Cell and Developmental Biology University of California at San Diego 9500 Gilman Dr. Natural Sciences Building 6105 La Jolla, CA 92093-0380 TEL (858) 534-6955 LAB (858) 822-4658 FAX (858) 822-5740 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080722/e1750f6b/attachment.html From thirumal from cshl.edu Wed Jul 23 17:46:42 2008 From: thirumal from cshl.edu (Thirumalai, Vatsala) Date: Wed Jul 23 17:51:16 2008 Subject: [Zbrafish] Physiological Saline References: Message-ID: <18C77A9DEE192D479EB353698DA15717100A0B@mailbox14.cshl.edu> Hi, Jason! I use saline with the following composition in mM: 134 NaCl; 2.9 KCl; 1.2 MgCl2; 10 HEPES; 10 Glucose; 2.1 CaCl2. I adjust the pH to 7.8 with 10N NaOH and the osmolality should be 290 mmol/Kg. You can find this recipe in papers from the Drapeau and the Fetcho labs as well as my own. Let me know if you have any more questions. Best, Vatsala Thirumalai. ________________________________ From: zbrafish-bounces@oat.bio.indiana.edu on behalf of Leviathan Sent: Tue 7/22/2008 9:25 PM To: bionet-organisms-zebrafish@moderators.isc.org Subject: [Zbrafish] Physiological Saline Hi everyone, I am hoping that some of the physiologists out there may be able to help me here. I have just had a researcher here in adelaide approach me inquiring as to whether she could do some physiological studies on the zebrafish. Considering past researchers here have all been a twist of the molecular and developmental biology this request is all very new for me. I think that what she is proposing is grand, but she asked what the composition of the physiological saline she should use for the tissue experiments should be to mimic the internal environment of a zebrafish? i have no idea, and thought i'd ask you all, to see if anyone out there may know. if anyone can offer assistance that would be excellent :O) many thanks, jason _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From ucgahar from ucl.ac.uk Thu Jul 24 07:13:31 2008 From: ucgahar from ucl.ac.uk (carole wilson) Date: Thu Jul 24 11:47:54 2008 Subject: [Zbrafish] Zebrafish Line Information Message-ID: Dear all, In response to the continued problems of importing fish, we have finally managed to get the Zebrafish line database online, the hope is that it will house all the lines that the community is willing to share with others in the UK and therefore alleviating some of the importing problems. I have already started by uploading some of the lines that UCL holds in its stock centre and are available to the community and I would encourage everyone else who has lines to share to also upload them. Please can you send this information on to anyone who might be interested and who may have been left off of this list Hopefully this will be a useful resource. The database can be found at http://www.ucl.ac.uk/zebrafish-group/ fishfacility/index.php Carole Wilson Fish Facility Manager University College London ucgahar@ucl.ac.uk -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080724/b600f341/attachment.html From gottalovelife from gmail.com Fri Jul 25 10:55:28 2008 From: gottalovelife from gmail.com (Kim) Date: Fri Jul 25 11:24:26 2008 Subject: [Zbrafish] Fungizone Message-ID: <2219bc1b-b8ab-407b-a637-9a794caff0a2@b30g2000prf.googlegroups.com> Has anyone ever used fungizone to prevent the growth of fungus on their zebrafish embryos? I am running toxicity tests and am not sure if the growth on the bottom of my plates is due to my toxicant or fungus and would like to eliminate any chance of it being fungus. If anyone has information on what concentration I should be using and at what stage I should add it to the water. Cheers! From biesemei from purdue.edu Fri Jul 25 11:44:45 2008 From: biesemei from purdue.edu (djbies) Date: Fri Jul 25 11:48:29 2008 Subject: [Zbrafish] BDNF antibody Message-ID: <03618666-edeb-44e1-8c33-af05e85274a7@n33g2000pri.googlegroups.com> Does anyone know of a BDNF antibody, preferably monoclonal, that works in zebrafish? From kbaden from gmail.com Fri Jul 25 11:48:02 2008 From: kbaden from gmail.com (Katy) Date: Fri Jul 25 11:48:49 2008 Subject: [Zbrafish] Re: ZebraFish Skin Histology References: Message-ID: <6818e34f-c85f-4247-9d4f-920cb704a13c@j1g2000prb.googlegroups.com> On Jul 21, 6:50 pm, alzaidan.abdul...@gmail.com wrote: > Hi, > > I’m currently studying the effect of contaminants on the skin of > Zebrafish through Histology. The problem I’m facing while processing > the tissue is after doing so the skin always detaches from the muscle. > The fixative used is 4% NBF for 24 hrs. And I’m only removing the skin > attached with muscle, also tried to cut a TS section (Dorsal to > Ventral through the vertebra) but similar effect. The thickness used > on microtome is 5um. > > Any help would be appreciated > > Abdullah Al-Zaidan Hello, Are you looking at H&E stained sections? At the ZIRC we use Dietrich's fixative for all H&E specimens. I've pasted our fixation protocol with a recipe for Dietrich's fixative below. Hope this helps. Katy Murray, DVM, Ph.D. Zebrafish International Resource Center * Fixation Protocol for Histopathology Specimens * Prepare Dietrich's Fixative (100 ml) as follows. Store fixative at room temperature. 30 ml Ethanol (95%) 10 ml Formalin (Formaldehyde 37% solution, histological grade, contains 10-15% methanol, Sigma # F1635) 2 ml Glacial Acetic Acid 58 ml Distilled Water * Euthanatize fish with an overdose of MS-222 (tricaine methane sulfonate). * Carefully slit open the body cavity along the belly, trying not to disrupt internal organs. * Use 10-15 ml of fixative per 1-2 fish in a sealable container. The volume of fixative should be at least 10 times the volume of the specimen. * Store and ship fixed fish in Dietrich's fixative. Fish can be stored for several weeks in Dietrich's fixative. From finchg from ohsu.edu Fri Jul 25 12:32:40 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Fri Jul 25 12:34:58 2008 Subject: [Zbrafish] Re: Fungizone References: Message-ID: On Jul 25, 8:55 am, Kim wrote: > Has anyone ever used fungizone to prevent the growth of fungus on > their zebrafish embryos? I am running toxicity tests and am not sure > if the growth on the bottom of my plates is due to my toxicant or > fungus and would like to eliminate any chance of it being fungus. If > anyone has information on what concentration I should be using and at > what stage I should add it to the water. > > Cheers! I am not familiar with fungizone. Many labs add methylene blue to thier embryo media to control fungus. Our lab uses at 0.02%. This ususally gets rid of any sign of fungus though it colors the water slightly blue. VWR Cat. No.VW6276-0 From jalexyu from gmail.com Sat Jul 26 01:26:02 2008 From: jalexyu from gmail.com (movingfish) Date: Mon Jul 28 11:52:18 2008 Subject: [Zbrafish] Zebrafish training session Message-ID: Does anyone know where i can get basic training on zebrafish development and experimental protocol? From M.Wiweger from lumc.nl Mon Jul 28 11:47:07 2008 From: M.Wiweger from lumc.nl (M.Wiweger@lumc.nl) Date: Mon Jul 28 16:10:46 2008 Subject: [Zbrafish] jek mutant Message-ID: Hello, I am interested in mutants with skeletal defects. Does anyone have jekyll (UDP-glucose dehydrogenase) mutant in stock and could send me some bleached eggs please? With kind regards, Malgorzata ________________________ Dr. Malgorzata Wiweger Leiden University Medical Center, Department of Pathology, L1-Q, Po Box 9600, 2300 RC Leiden, The Netherlands tel: +31 71 526-65-96 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080728/875647e2/attachment.html From mwinandy from ulg.ac.be Mon Jul 28 03:04:32 2008 From: mwinandy from ulg.ac.be (Marie Winandy) Date: Tue Jul 29 12:16:43 2008 Subject: [Zbrafish] Re: Fungizone In-Reply-To: <200807251704.m6PH4EO05884@net.bio.net> References: <200807251704.m6PH4EO05884@net.bio.net> Message-ID: <200807281004320906.00456B6C@smtp.ulg.ac.be> Dear Kim and dear all, We usually use a mix of penicillin-streptomycin-amphotericin 100x concentrated (Invitrogen antibiotic-antimycotic cat nr 15240). This mixture really helps in controlling bacterial and fungal infection, but I must add that you have to use very clean fish or egg water, contained in clean beakers etc, to decrease the contaminant level. Best reMarie Winandy, PhD Université de Liège GIGA B34 - Zebrafish Platform Avenue de l'Hôpital 1 B-4000 Liège - Sart Tilman Belgique Tél: +32.4.366.99.71 +32.4.366.33.38 +32.476.97.25.33 Fax: +32.4.366.41.98 email : mwinandy@ulg.ac.be or zebrafish.giga@ulg.ac.be http://www.giga.ulg.ac.be/extranet/services.htmgards, From sspinette from ric.edu Tue Jul 29 08:08:49 2008 From: sspinette from ric.edu (sarah) Date: Tue Jul 29 12:17:04 2008 Subject: [Zbrafish] Re: Zebrafish training session References: Message-ID: <98a193cb-96b2-40d6-8881-20922eab4d1f@e39g2000hsf.googlegroups.com> On Jul 26, 2:26?am, movingfish wrote: > Does anyone know where i can get basic training on zebrafish > development and experimental protocol? MBL offers intense summer courses although the price of tuition and room and board is very high! http://www.mbl.edu/education/courses/special_topics/zebra.html I am looking for something like this myself - basic hands on experimental techniques etc. From caroline.parkin from sheffield.ac.uk Wed Jul 30 04:46:58 2008 From: caroline.parkin from sheffield.ac.uk (Caroline Parkin) Date: Wed Jul 30 11:45:58 2008 Subject: [Zbrafish] Re: Zebrafish training In-Reply-To: <200807281703.m6SH35O27581@net.bio.net> References: <200807281703.m6SH35O27581@net.bio.net> Message-ID: <310F7735-43E7-4E4B-B2C3-1C56864917DE@shef.ac.uk> Hi, Please can you tell us where you are located? Within the UK/Europe it may be possible to get training here at the University of Sheffield (this is something new we are considering offering, depending on the amount of interest - feedback on this idea would be welcome). best wishes, Caroline caroline.parkin@shef.ac.uk On 28 Jul 2008, at 18:03, zbrafish-request@oat.bio.indiana.edu wrote: > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Zebrafish training session (movingfish) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 25 Jul 2008 23:26:02 -0700 (PDT) > From: movingfish > Subject: [Zbrafish] Zebrafish training session > To: bionet-organisms-zebrafish@moderators.isc.org > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Does anyone know where i can get basic training on zebrafish > development and experimental protocol? > > > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 38, Issue 16 > **************************************** From bdschyth from gmail.com Thu Jul 31 05:11:22 2008 From: bdschyth from gmail.com (bdschyth) Date: Thu Jul 31 10:58:27 2008 Subject: [Zbrafish] Re: Zebrafish training References: <200807281703.m6SH35O27581@net.bio.net> Message-ID: On Jul 30, 11:46?am, Caroline Parkin wrote: > Hi, > Please can you tell us where you are located? Within the UK/Europe it ? > may be possible to get training here at the University of Sheffield ? > (this is something new we are considering offering, depending on the ? > amount of interest - feedback on this idea would be welcome). > best wishes, > Caroline > caroline.par...@shef.ac.uk > > On 28 Jul 2008, at 18:03, zbrafish-requ...@oat.bio.indiana.edu wrote: > > > > > Send Zbrafish mailing list submissions to > > ? ?zbraf...@net.bio.net > > > To subscribe or unsubscribe via the World Wide Web, visit > > ? ?http://www.bio.net/biomail/listinfo/zbrafish > > or, via email, send a message with subject or body 'help' to > > ? ?zbrafish-requ...@net.bio.net > > > You can reach the person managing the list at > > ? ?zbrafish-ow...@net.bio.net > > > When replying, please edit your Subject line so it is more specific > > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > > ? 1. Zebrafish training session (movingfish) > > > ---------------------------------------------------------------------- > > > Message: 1 > > Date: Fri, 25 Jul 2008 23:26:02 -0700 (PDT) > > From: movingfish > > Subject: [Zbrafish] Zebrafish training session > > To: bionet-organisms-zebraf...@moderators.isc.org > > Message-ID: > > ? ? > > Content-Type: text/plain; charset=ISO-8859-1 > > > Does anyone know where i can get basic training on zebrafish > > development and experimental protocol? > > > ------------------------------ > > > _______________________________________________ > > Zbrafish mailing list > > Zbraf...@net.bio.net > >http://www.bio.net/biomail/listinfo/zbrafish > > > End of Zbrafish Digest, Vol 38, Issue 16 > > ****************************************- Hide quoted text - > > - Show quoted text - Dear Caroline Depending on the issues covered I would also be interested in such a course closer to Denmark where I am based. I work on using siRNAs in rainbow trout and recently also on miRNA regulation during viral infection. Brian From cayuelamarialuisa from gmail.com Thu Jul 31 03:47:23 2008 From: cayuelamarialuisa from gmail.com (MariaLuisa Cayuela) Date: Thu Jul 31 10:58:57 2008 Subject: [Zbrafish] Alternative to gene Tools In-Reply-To: <52ac5730-0d54-4867-937c-088740bfd5a9@m3g2000hsc.googlegroups.com> References: <52ac5730-0d54-4867-937c-088740bfd5a9@m3g2000hsc.googlegroups.com> Message-ID: <84e45b690807310147o6c4dfc54x3269d2aba9875e99@mail.gmail.com> hello I think there is a altenative the Grip you can purchanse from Active Motif (Belgium). have a nice day 2008/7/15, hernoc@gmail.com : > Hi Neters, does anyone know of an alternative commercial and reliable > source for morpholinos? GeneTools is becoming a real pain in the *** > to work with. Thanks a lot, > Hernan Lopez-Schier.- > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20080731/7c4b9208/attachment.html From jan.gebauer from uni-koeln.de Thu Jul 31 03:24:45 2008 From: jan.gebauer from uni-koeln.de (Jan Gebauer) Date: Thu Jul 31 10:59:20 2008 Subject: [Zbrafish] Zebrafish cell lines? Message-ID: Hi Zebrafish Community, I'm looking for an easy-to-handle / easy-to-get zebrafish cell line for a colleague of mine. He'd like to have a look at its glycosylation profile so I guess every cell type will do. Does anyone know an established zebrafish cell line and where I can get it? Thanks, Jan -- Dr. Jan Gebauer phone: +49 221 4786990 | fax: +49 221 4786977 University of Cologne, Medical Faculty Research group: Dr. Raimund Wagener Head of Department: Prof. Mats Paulsson Institute for Biochemistry II, Joseph-Stelzmann-Strasse 52, D50931 Cologne From mena22787 from excite.com Thu Jul 31 11:43:57 2008 From: mena22787 from excite.com (Serena) Date: Thu Jul 31 12:00:09 2008 Subject: [Zbrafish] cryosections and staining Message-ID: <7880a736-c83c-4f31-a276-0acda19e62c0@m3g2000hsc.googlegroups.com> Hi all! I'm an undergrad at Rhode Island College and was wondering if anyone has any protocols/helpful advice on performing immunohistochemistry or in situ hybridization on zebrafish cryosections? We have divided whole adult fish into head/heart, body, and end/tail sections and frozen them in tissue freezing medium. We hope to locate a specific cytoplasmic protein. Thanks! From lampe1 from umbc.edu Thu Jul 31 12:19:05 2008 From: lampe1 from umbc.edu (lampe1) Date: Thu Jul 31 12:23:21 2008 Subject: [Zbrafish] help with parasite infecting embryos Message-ID: <1166b243-2f21-442e-8542-268a72f1a099@l42g2000hsc.googlegroups.com> Hello! In our zebrafish facility, we frequently have embryos infected with some sort of parasite. It looks like little filaments of cotton which surround the embryo. When this happens, the embryos usually die within 24-72 hours. Usually bleaching (according to ZFIN protocol--0.1mL 5% sodium hypochlorite in 170mL system water, bleach 5 min, system water 5 min, bleach 5 min, rinse 2+ times in system water) and replacing the water stops this and saves the embryos. Recently, this parasite has been worse (likely due to our UV sterilizer being broken for ~1 week). One person in the lab bleached the embryos and rinsed them several times and yet they still all died and were infected with this parasite. Does anyone have any idea what this parasite may be? I am having trouble since I have no idea what kind of organism it even is. Does anyone have any ideas for other ways to bleach/otherwise save embryos? I appreciate any help! Thanks, Rebecca Lampe Fish Care Technician UMBC Department of Biology lampe1@umbc.edu From SarBOgden from gmail.com Thu Jul 31 14:15:00 2008 From: SarBOgden from gmail.com (SarBOgden@gmail.com) Date: Thu Jul 31 14:24:43 2008 Subject: [Zbrafish] Zebrafish food Message-ID: <9f9fb284-c919-4228-b55d-065ce97c4679@56g2000hsm.googlegroups.com> Hello, In my facility we've been feeding Rangen Trout and Salmon starter. We've been using the same bag that we've kept in the fridge for 5 years. Does anyone use this food and know if it lasts this long? We are looking to order new food but are looking for other options. Any suggestions would be greatly appreciated. Thanks, Sarah Lab Technician UMBC sarbogden@gmail.com From Christian.Lawrence from childrens.harvard.edu Thu Jul 31 14:57:33 2008 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Thu Jul 31 15:28:14 2008 Subject: [Zbrafish] Zebrafish food Message-ID: <1D117326B8F30943ACD9A47E1BEE2941C4B57F@CHEXV2.CHBOSTON.ORG> The shelf life on processed feeds is generally no more than 3 months, if they are stored correctly (cool and dry). If you don't store properly, it is less than that. There are several processed feeds out there that support good production for zebrafish, although none have been demonstrated to be nutritionally complete. We use Lansy NRD 400-600 pellet by INVE aquaculture. This works best when used as part of a diet that includes Artemia. It costs approximately 7-10$/kg. Chris ----- Original Message ----- From: zbrafish-bounces@oat.bio.indiana.edu To: bionet-organisms-zebrafish@moderators.isc.org Sent: Thu Jul 31 15:15:00 2008 Subject: [Zbrafish] Zebrafish food Hello, In my facility we've been feeding Rangen Trout and Salmon starter. We've been using the same bag that we've kept in the fridge for 5 years. Does anyone use this food and know if it lasts this long? We are looking to order new food but are looking for other options. Any suggestions would be greatly appreciated. Thanks, Sarah Lab Technician UMBC sarbogden@gmail.com _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish The information transmitted in this electronic communication is intended only for the person or entity to whom it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon this information by persons or entities other than the intended recipient is prohibited. If you received this information in error, please contact the Compliance HelpLine at 800-856-1983 and properly dispose of this information. From finchg from ohsu.edu Thu Jul 31 16:02:17 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Thu Jul 31 17:29:57 2008 Subject: [Zbrafish] Re: help with parasite infecting embryos References: Message-ID: <0764e5ff-e9d6-43d3-806e-5826324fd4c5@u6g2000prc.googlegroups.com> On Jul 31, 10:19 am, lampe1 wrote: > Hello! > > In our zebrafish facility, we frequently have embryos infected with > some sort of parasite. It looks like little filaments of cotton which > surround the embryo. When this happens, the embryos usually die within > 24-72 hours. Usually bleaching (according to ZFIN protocol--0.1mL 5% > sodium hypochlorite in 170mL system water, bleach 5 min, system water > 5 min, bleach 5 min, rinse 2+ times in system water) and replacing the > water stops this and saves the embryos. > > Recently, this parasite has been worse (likely due to our UV > sterilizer being broken for ~1 week). One person in the lab bleached > the embryos and rinsed them several times and yet they still all died > and were infected with this parasite. > > Does anyone have any idea what this parasite may be? I am having > trouble since I have no idea what kind of organism it even is. Does > anyone have any ideas for other ways to bleach/otherwise save embryos? > > I appreciate any help! > > Thanks, > > Rebecca Lampe > Fish Care Technician > UMBC Department of Biology > lam...@umbc.edu I'm assuming the infection is occuring in the first few days when the fish are in petri dishes. The filaments sound like fungus. Fungus growing in the dishes is fairly common and will kill the embryos. See the discussion regarding Fungizone from a few days ago as it contains a variety of recommendations. From finchg from ohsu.edu Thu Jul 31 16:07:29 2008 From: finchg from ohsu.edu (finchg@ohsu.edu) Date: Thu Jul 31 17:30:33 2008 Subject: [Zbrafish] Re: cryosections and staining References: Message-ID: http://zebrafish.org/zirc/documents/protocols.php http://www.fhcrc.org/science/labs/moens/Protocols/ From dgw5079 from u.washington.edu Thu Jul 31 17:47:33 2008 From: dgw5079 from u.washington.edu (David G. White) Date: Thu Jul 31 17:50:16 2008 Subject: [Zbrafish] help with parasite infecting embryos In-Reply-To: <1166b243-2f21-442e-8542-268a72f1a099@l42g2000hsc.googlegroups.com> Message-ID: The little filaments are most likely fungal hyphae, saprolegnia etc and would come more problematic with a broken UV. One method of treating the embryos would be using methylene blue, which can be found at any pet store. Add enough to your egg medium until it turns blue. There is a few draw backs to methylene blue. It will kill bacterial beds so do not want to use it on any kind of water recirculating nursery with a biofilter. Also it can stain cells blue. The fungal spores will exist in your fish system water even with a fully-functioning UV. If you use spawning cages you can try setting up your fish later in the day, so that you reduce the amount of fecal matter deposited in the cages with the eggs. We collect the eggs promptly in the morning. Pour the eggs through a 200 um screen and rinse them with fresh egg water in to a petri dish. Under the scope remove all debris and any infertile eggs. Good eggs should be perfectly round with a dark round yolk. If your not certain if the egg is good or not I would remove it because a few bad eggs can spoil the whole lot. The next day look over the eggs again, and clean if necessary. We do not use methylene blue. You also might want to increase maintenance on your UV, replacing the bulb and cleaning the quartz sleeve or perhaps up-sizing it. Good luck David G White Research Coordinator H225 Zebrafish Lab University of Washington Department of Biological Structure HSB G514 Box 357420 1959 NE Pacific Street Seattle, WA 98195-7420 Tel. 206-685-7512 FAX 206-543-1524 On Thu, 31 Jul 2008, lampe1 wrote: > Hello! > > In our zebrafish facility, we frequently have embryos infected with > some sort of parasite. It looks like little filaments of cotton which > surround the embryo. When this happens, the embryos usually die within > 24-72 hours. Usually bleaching (according to ZFIN protocol--0.1mL 5% > sodium hypochlorite in 170mL system water, bleach 5 min, system water > 5 min, bleach 5 min, rinse 2+ times in system water) and replacing the > water stops this and saves the embryos. > > Recently, this parasite has been worse (likely due to our UV > sterilizer being broken for ~1 week). One person in the lab bleached > the embryos and rinsed them several times and yet they still all died > and were infected with this parasite. > > Does anyone have any idea what this parasite may be? I am having > trouble since I have no idea what kind of organism it even is. Does > anyone have any ideas for other ways to bleach/otherwise save embryos? > > I appreciate any help! > > Thanks, > > Rebecca Lampe > Fish Care Technician > UMBC Department of Biology > lampe1@umbc.edu > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > From RachelRaynes from gmail.com Thu Jul 31 20:16:41 2008 From: RachelRaynes from gmail.com (RachelRaynes@gmail.com) Date: Mon Aug 4 11:34:18 2008 Subject: [Zbrafish] Swarming zebrafish embryo parasites Message-ID: <9d97a969-187c-46c4-a594-5ea426f8b81f@56g2000hsm.googlegroups.com> While performing maintenance on a clutch of 48 hour zebrafish embryos, I came across one with a swarm of parasites within its chorion. All the other embryos in the clutch were fine. I examined the embryo 5 hours later and it was almost completely devoured. I assume that this is a parasite that the mother passed on? Unfortunately, the cross was albino stock and the individual fish was put back into a group tank after breeding. What were those things and what action if any needs to be taken? If anyone would like a picture or video of the embryo I’d be happy to send one in hopes of identifying the parasite.