From chubbyjayu from gmail.com Sun Sep 6 23:11:26 2009 From: chubbyjayu from gmail.com (chubbyjayu) Date: Tue Sep 8 11:39:07 2009 Subject: [Zbrafish] On raising Zebrafish larvae Message-ID: <5a8c2d96-b211-4e35-a976-9298800518e4@v15g2000prn.googlegroups.com> Hello all! I have been trying to raise zebrafish larvae in the lab unsuccessfully. The problem is that the larvae tend to die 12 - 14 days post fertilization and there seems to be no logical cause for the death. I check the paramecium stocks for coleps everyday, but see none. We have even tried raising the zebrafish larvae in embryo medium (made as per the zebrafish book), but even then we see that the larvae die out within 14 days. Could you please help me figure out where I am going wrong? Jayasree From naruse from nibb.ac.jp Mon Sep 7 03:08:08 2009 From: naruse from nibb.ac.jp (Kiyoshi Naruse) Date: Tue Sep 8 11:39:37 2009 Subject: [Zbrafish] dead line of NIBB practical cource Message-ID: <4AA4BF68.1090505@nibb.ac.jp> Dear all, Application deadline for practical course "Developmental Genetics ofZebrafish and Medaka $B-7(B " is October 5th, 2009 This course cover the making transgenic medaka with BAC homologous recombination technology and sperm cryopreservation for long time storage of TG and mutant lines. International flight fare (not domestic) and accommodation costs will be funded for foreign graduate students. please visit the website for NIBB international practical course "http://www.nibb.ac.jp/course/index.html" We are waiting for your application. All the best ---------------------------------------- Kiyoshi NARUSE Ph.D. National Institute for Basic Biology, Laboratory of Bioresources, Nishigonaka 38, Myodaiji, Okazaki 444-8585, Aichi, Japan TEL: 0564-55-7581$B!!(BTEL/FAX: 0564-55-7580 email: naruse@nibb.ac.jp ----------------------------------------------------- From finchg.ohsu.edu from gmail.com Tue Sep 8 15:38:51 2009 From: finchg.ohsu.edu from gmail.com (finchg@ohsu.edu) Date: Tue Sep 8 15:41:40 2009 Subject: [Zbrafish] Re: Question about Personnel to Rack/Tank Ratio References: Message-ID: On Aug 30, 9:32?am, Caroline wrote: > Hello All, > > ? ? ?I'm the Zebrafish Technician for a facility with 18 racks (~1100 > tanks), and 3 more are scheduled to be installed tomorrow ( for a > total of 4 separate recirculating systems). Currently, I'm the only > staff member who cares for the fish and facility, and I'm quite > concerned that I will not be able to keep up with the workload. Since > aquatic model organisms aren't as strictly regulated or as well- > established as the murine species, I've not been able to find any > information regarding the "typical" job description of the Zebrafish > Tech, how other institutions delegate fish facilities duties, and how > many people they employ to cover these tasks. So, I was hoping to get > some feedback on this matter from other members of the zebrafish > community for a means of comparison, and hopefully some information to > take to my supervisors. > > Thanks so much > Caroline Caroline, We have two full time fish techs in our lab caring for about 16 AHAB-style racks of fish and additionally raising fry, culturing para/ shrimp, doing extensive test crossing, and fin-clipping. You are correct that there is scant information with regards to "typical" job responsibilities for a fish tech. I think that job description varies a lot from facility to facility, sometimes a tech might be responsible for crossing lines, raising fry, ordering supplies, doing janitorial, washing cages/tanks, fixing broken system components, etc. whereas in other facilities they might be just feeding (though this is probably rare). In many cases the PI may have little or no sense of the work load involved in caring for the fish, therefore I am sympathetic to your plea for some hard data to take to them. Don't be afraid to ask for help or to let your superiors know if your work load is unreasonable. If you work at an undergrad university, many facilities use work-study students as aids to their fish techs. Remember, as a fish tech your primary responsibility is to keep the fish healthy (which might mean asking for additional help). Good Luck From finchg.ohsu.edu from gmail.com Tue Sep 8 15:52:22 2009 From: finchg.ohsu.edu from gmail.com (finchg@ohsu.edu) Date: Tue Sep 8 16:17:29 2009 Subject: [Zbrafish] Re: On raising Zebrafish larvae References: Message-ID: On Sep 6, 9:11?pm, chubbyjayu wrote: > Hello all! > I have been trying to raise zebrafish larvae in the lab > unsuccessfully. The problem is that the larvae tend to die 12 - 14 > days post fertilization and there seems to be no logical cause for the > death. > I check the paramecium stocks for coleps everyday, but see none. We > have even tried raising the zebrafish ?larvae in embryo medium (made > as per the zebrafish book), but even then we see that the larvae die > out within 14 days. > Could you please help me figure out where I am going wrong? > Jayasree Jayasree, Hopefully these pointers will be helpful. Even using E3 the fish are going to need a gradual water change. Some facilities put the babies on an overnight drip/trickle which is turned off in the day. When the fish are dying at 12-14 days, this usually indicates that they are not getting enough to eat (I've been told the yolk runs out around day 10). In my experience fish do best with an extreme concentrations of paramecia. You want the it too look like a paramecia blizzard in the tank if you can. This is why using a small volume tank or beaker (greater para. conc.) works best for the larvae. If the fish are well fed you can see their bellies bulged out even without magnification. It seems to be nearly impossible to "overfeed" babies (of course rotting food in the tank is a no-no, but this doesn't seem to happen easily with para). Hope this helps. From Christian.Lawrence from childrens.harvard.edu Tue Sep 8 21:50:41 2009 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Wed Sep 9 10:34:27 2009 Subject: [Zbrafish] On raising Zebrafish larvae In-Reply-To: <5a8c2d96-b211-4e35-a976-9298800518e4@v15g2000prn.googlegroups.com> Message-ID: In all likelihood, the fish are basically starving. At 28C, they start exogenous feeding at 5 days, but also continue to derive nutritional benefit from the yolk sac, but this is generally gone by day 7. If you don't give them any food at all, they will starve at day 10 or so. No matter how dense, standard, bacteria-fed Paramecium does not meet the considerable nutritional demands of the fish beyond the first few days of feeding. If you do not present the larvae with something else with a better nutritional profile (higher protein, and specific lipid content) at this point, they will starve. If all of your fish die by 12-14 days, then that is likely what is happening. The Paramecium typically gets them out a day or two longer than the 10 day mark. Even if you manage to keep them alive beyond that, they won't grow until you give them something with an adequate nutritional profile. You're just prolonging the starvation. If you start feeding Paramecium at day 5 (as soon as they are swimming), you should be presenting them with another item, in small, frequent applications, whether it be first stage Artemia nauplii or a processed larval feed, by 7-9 days. Once you begin do this, you should put them on flow at a slow drip to maintain stable (but not necessarily pristine) water quality. Once it becomes apparent that they are feeding on the new items (full guts, rapid growth), you can stop the paramecium applications, increase the amounts of the new feed per feeding, and slowly increase flow rates as the fish grow. The weaning from a low nutritional quality first feed like Paramecium to something better is the key to success in larval rearing. If you don't do it correctly or well, your survival rates will always be poor. Chris On 9/7/09 12:11 AM, "chubbyjayu" wrote: Hello all! I have been trying to raise zebrafish larvae in the lab unsuccessfully. The problem is that the larvae tend to die 12 - 14 days post fertilization and there seems to be no logical cause for the death. I check the paramecium stocks for coleps everyday, but see none. We have even tried raising the zebrafish larvae in embryo medium (made as per the zebrafish book), but even then we see that the larvae die out within 14 days. Could you please help me figure out where I am going wrong? Jayasree _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. From claudia_hohn from hotmail.com Wed Sep 9 08:29:02 2009 From: claudia_hohn from hotmail.com (Claudia) Date: Wed Sep 9 10:58:04 2009 Subject: [Zbrafish] Re: On raising Zebrafish larvae References: Message-ID: <43ce0df4-e122-4c24-9eb1-18b5286f20c4@t2g2000yqn.googlegroups.com> Hi, if you have static tanks you need to change water twice a day and remove uneaten food. Regular system water works just fine if you have fungus problems add Fungus Eliminator to the water. Also we start feeding a mix of Paramecium and Artemia starting 7dph. You can see our detailed methods at http://www.cvm.msstate.edu/basic_sciences/faculty/petrie-hanson_lora.html click on link zebrafish maintenance. Hope this helps, Claudia From rburdine from Princeton.EDU Wed Sep 9 10:52:30 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Thu Sep 10 11:37:02 2009 Subject: [Zbrafish] On raising Zebrafish larvae In-Reply-To: References: <5a8c2d96-b211-4e35-a976-9298800518e4@v15g2000prn.googlegroups.com> Message-ID: <790DAC09623D0A47ACDB126E3C52D8B62DF1CB@MBCLUSTER.pu.win.princeton.edu> I would like to add that we raise our larvae on dry food until day 20 or so. We do not use paramecium. Around day 20 we start feeding brine shrimp in addition to the dry food.. We have had good luck with AZ100 and BioKyowa for fish 5-10 days old. We have used various sized Zeigler AP100 dry food for 10-30 days. We typically get survival over 80% depending on the strain. Becky --------------------------------------------------- Rebecca D. Burdine, Ph.D. Assistant Professor Dept. of Molecular Biology Princeton University Washington Road Mof 433 Princeton, NJ 08544? ? Phone:?(609) 258-7515 Fax: (609) 258-6730 Email: rburdine@princeton.edu Admin Assistant: Anna Schmedel (609) 258-5028 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of Lawrence, Christian > Sent: Tuesday, September 08, 2009 10:51 PM > To: chubbyjayu; bionet-organisms-zebrafish@moderators.isc.org > Subject: Re: [Zbrafish] On raising Zebrafish larvae > > In all likelihood, the fish are basically starving. > > At 28C, they start exogenous feeding at 5 days, but also continue to derive > nutritional benefit from the yolk sac, but this is generally gone by day 7. If you > don't give them any food at all, they will starve at day 10 or so. > > No matter how dense, standard, bacteria-fed Paramecium does not meet the > considerable nutritional demands of the fish beyond the first few days of feeding. > If you do not present the larvae with something else with a better nutritional > profile (higher protein, and specific lipid content) at this point, they will starve. If > all of your fish die by 12-14 days, then that is likely what is happening. The > Paramecium typically gets them out a day or two longer than the 10 day mark. > Even if you manage to keep them alive beyond that, they won't grow until you > give them something with an adequate nutritional profile. You're just prolonging > the starvation. > > If you start feeding Paramecium at day 5 (as soon as they are swimming), you > should be presenting them with another item, in small, frequent applications, > whether it be first stage Artemia nauplii or a processed larval feed, by 7-9 days. > Once you begin do this, you should put them on flow at a slow drip to maintain > stable (but not necessarily pristine) water quality. Once it becomes apparent that > they are feeding on the new items (full guts, rapid growth), you can stop the > paramecium applications, increase the amounts of the new feed per feeding, > and slowly increase flow rates as the fish grow. > > The weaning from a low nutritional quality first feed like Paramecium to > something better is the key to success in larval rearing. If you don't do it correctly > or well, your survival rates will always be poor. > > Chris > > > > > > > On 9/7/09 12:11 AM, "chubbyjayu" wrote: > > Hello all! > I have been trying to raise zebrafish larvae in the lab > unsuccessfully. The problem is that the larvae tend to die 12 - 14 > days post fertilization and there seems to be no logical cause for the > death. > I check the paramecium stocks for coleps everyday, but see none. We > have even tried raising the zebrafish larvae in embryo medium (made > as per the zebrafish book), but even then we see that the larvae die > out within 14 days. > Could you please help me figure out where I am going wrong? > Jayasree > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > > The information in this e-mail is intended only for the person to whom it is > addressed. If you believe this e-mail was sent to you in error and the e-mail > contains patient information, please contact the Partners Compliance HelpLine at > http://www.partners.org/complianceline . If the e-mail was sent to you in error > but does not contain patient information, please contact the sender and properly > dispose of the e-mail. > > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From jason.cockington from gmail.com Fri Sep 11 00:55:58 2009 From: jason.cockington from gmail.com (Leviathan) Date: Fri Sep 11 11:18:52 2009 Subject: [Zbrafish] Re: On raising Zebrafish larvae References: <5a8c2d96-b211-4e35-a976-9298800518e4@v15g2000prn.googlegroups.com> Message-ID: <9b67f0c5-14fd-4073-b475-d34515058a4a@e4g2000prn.googlegroups.com> Hi Jayasree, rest assured that you are not alone in your struggle for efficiently closing the life cycle of the zebrafish. There are many labs out there that use a variety of different methods for rearing their very young fish. If you haven't already, may i suggest that you become a member of the Zebrafish Husbandry Association. This is a world wide group dedicated to the promotion of better understanding the needs of zebrafish, and larval rearing just happens to be the flavour of the month at the moment. The ZHA recently organised a webinar presented by Isaac Adatto on the "Evaluation of Various Live Feeds and Feeding Regimes on Growth and Survival of Larval Zebrafish". If you can spare the time, it would be something you would find most interesting. you can find the ZHA here: http://www.zhaonline.org then if you want to find the webinar, look for isaac's talk in the ZHA Library. Jason From witmer.dane from gmail.com Fri Sep 11 13:42:04 2009 From: witmer.dane from gmail.com (bionet.organisms.zebrafish) Date: Fri Sep 11 13:43:05 2009 Subject: [Zbrafish] embedding embryos in plastic Message-ID: <4e732a7f-7392-45c4-b34e-b291530f326b@o13g2000vbl.googlegroups.com> Dear List, I plan to embed some embryos (5dpf) using technovit. I wanted to find out the best molds to use for this purpose, and whether any special chuck is needed for the microtome. Part numbers would be a plus. Thanks! Dane Witmer From busquet.francois from gmail.com Mon Sep 14 08:41:03 2009 From: busquet.francois from gmail.com (=?ISO-8859-1?Q?fran=E7ois?=) Date: Mon Sep 14 12:10:58 2009 Subject: [Zbrafish] breeding transgenic zebrafish References: Message-ID: Dear all, I would like to know if anybody has experience with this transgenic zebrafish line (BBD-T-010)? Moreover, are there disadvantages for breeding transgenic fishes? Is it more difficult than for the wild type? and where you can get them? looking forward for your replies thanks a lot francois From f.j.vaneeden from sheffield.ac.uk Tue Sep 15 03:42:40 2009 From: f.j.vaneeden from sheffield.ac.uk (freek) Date: Tue Sep 15 11:53:08 2009 Subject: [Zbrafish] GFP staining adult wax sections Message-ID: <3595785a-f51c-41b5-bd77-873cb26bb3df@g23g2000vbr.googlegroups.com> We are desparate for a "proven to work" protocol for GFP antibody staining that works on adult wax sections. I'll would be very grateful and offer a good bottle of wine for the first one that works, to be handed over on a future zf meeting From juantraverso from gmail.com Wed Sep 23 09:28:58 2009 From: juantraverso from gmail.com (Juan Traverso) Date: Wed Sep 23 11:27:17 2009 Subject: [Zbrafish] Morpholino oligos injection Message-ID: <77b3339f-9703-44ce-b502-d338f327881b@a6g2000vbp.googlegroups.com> Hello, I'd like to know if someone did found any differences in the phenotype and/or mortality rates obtained while injecting MOs either in the yolk or the 1-cell embryo itself?. I'm new to the group and I guess this may have been discussed before...Thanks in advance for your answers, Juan From robinhaw from gmail.com Wed Sep 23 14:18:31 2009 From: robinhaw from gmail.com (rhaw) Date: Wed Sep 23 14:33:49 2009 Subject: [Zbrafish] Reactome Pathway Database User Survey Message-ID: Reactome is committed to providing access to high-quality pathway information and helpful data analysis tools. With this in mind, we are actively soliciting comments from the research community in order to assess community needs. We are interested to hear about your experience with Reactome, and would like to know a bit about your background and research interests so that we can continue to improve the Reactome site and tools. You can access the survey at: http://tinyurl.com/l48zzq Thank you for taking part. Robin Haw Manager of Reactome Outreach Outreach [at] reactome.org http://www.reactome.org From karlstrom from bio.umass.edu Wed Sep 23 14:35:51 2009 From: karlstrom from bio.umass.edu (Rolf Karlstrom) Date: Wed Sep 23 14:40:39 2009 Subject: [Zbrafish] EvoDevo Job at UMass Amherst Message-ID: <5D9BA103-88A1-4C31-A635-D8EF361EC19A@bio.umass.edu> I'd like to alert folks to the following position that was advertised in the Sept. 18th issue of Science. The Department of Biology at the University of Massachusetts Amherst (www.bio.umass.edu/biology) invites applications for a tenure-track assistant professorship in the area of Evolutionary/Developmental Biology (Evo/Devo). The UMass Biology Department provides an interactive and broad research environment, with faculty research spanning levels of biological organization. Especially strong research clusters focus on the areas of Neural Developmental, Cell Biology, Plant Biology, Functional Morphology, and Evolution. Successful candidates will have a Ph.D., postdoctoral experience, and the potential to develop and maintain an extramurally funded research program. New faculty members will have the opportunity to participate in strong graduate training programs in Molecular and Cellular Biology (www.bio.umass.edu/mcb), Organismal and Evolutionary Biology (www.bio.umass.edu/oeb), Plant Biology (www.bio.umass.edu/ plantbio), and Neuroscience and Behavior (www.umass.edu/neuro). The candidate should have a strong record of research examining molecular/cellular mechanisms of development that are of fundamental importance for evolutionary biology. We are interested in candidates whose research addresses issues such as the origin and diversification of body plans, the functional divergence of regulatory pathways into novel developmental systems, or the evolution of body structures. Work with model systems, including nematodes, flies, or fish is encouraged, but outstanding applicants who work on other systems will also be considered. Successful candidates should compliment and bridge current departmental research strengths in neural development, evolutionary biology, and functional morphology. Evaluation of applications will begin on October 5, 2009 and continue until the position is filled. Position to be filled contingent upon University funding. Please send application materials to: Evo/Devo Search #R36699, Biology Department, Attn: Karen Nelson, 611 North Pleasant Street, University of Massachusetts, Amherst, MA 01003. Application materials should include a curriculum vitae, research plan, teaching statement, plus the names, phone numbers, and email addresses for 3 referees. 3 letters of recommendation can either be included in the packet or sent electronically to: evodevosearch@bio.umass.edu. The University of Massachusetts is an Affirmative Action/Equal Opportunity Employer. Women and members of minority groups are encouraged to apply. The Biology Department is aggressive in its efforts to hire candidates who will enhance the diversity and general balance of the faculty and the sciences. ? Rolf Karlstrom Chair, Department of Biology University of Massachusetts Amherst, MA 01003 Office: 413-577-3448 Lab: 413-577-3456 karlstrom@bio.umass.edu -------------- next part -------------- Skipped content of type multipart/mixed From jps from stowers.org Thu Sep 24 11:33:28 2009 From: jps from stowers.org (stowersjps) Date: Thu Sep 24 11:36:31 2009 Subject: [Zbrafish] Post-anesthesia procedures for zebrafish Message-ID: What are other facilities’ post-anesthesia procedures for zebrafish? Why are these procedures utilized? From chubbyjayu from gmail.com Fri Sep 25 10:34:38 2009 From: chubbyjayu from gmail.com (chubbyjayu) Date: Fri Sep 25 11:50:16 2009 Subject: [Zbrafish] Re: On raising Zebrafish larvae References: <5a8c2d96-b211-4e35-a976-9298800518e4@v15g2000prn.googlegroups.com> Message-ID: Hi there! Heartfelt thanks to all who replied to my post with hints. Just an update, the fry are doing fine at 21 days post fertilization. The mortality rate too, has dropped as now I introduce the fry to macerated fish food at day 5 and by day 6 they seem to be eating it. (Of course, paramecium is added for the smaller ones, and artemia is given to the older fry) Thanks once again! Cheerio Jayasree From mwinandy from ulg.ac.be Fri Sep 25 02:05:16 2009 From: mwinandy from ulg.ac.be (Marie Winandy) Date: Fri Sep 25 11:50:35 2009 Subject: [Zbrafish] post anesthesia procedure In-Reply-To: <200909241703.n8OH3ak24394@net.bio.net> References: <200909241703.n8OH3ak24394@net.bio.net> Message-ID: <4ABC6BAC.6030903@ulg.ac.be> An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090925/842c455c/attachment.html -------------- next part -------------- A non-text attachment was scrubbed... Name: mwinandy.vcf Type: text/x-vcard Size: 398 bytes Desc: not available Url : http://www.bio.net/bionet/mm/zbrafish/attachments/20090925/842c455c/mwinandy.bin From richard.kollmar from downstate.edu Tue Sep 29 20:38:56 2009 From: richard.kollmar from downstate.edu (Richard Kollmar) Date: Wed Sep 30 11:35:23 2009 Subject: [Zbrafish] Cage washers and detergents Message-ID: <095B30DB-193D-4C1D-886F-BD358DD76B2F@downstate.edu> Dear All: I would like to put used tanks, nets, etc. after mechanical cleaning through the cage washer of our animal facility for sanitization. I am worried, though, about the detrimental effects of detergents on fish health. My previous institution had enough fish and frog labs to make it worthwhile draining the cage washer once a week to sanitize our equipment without detergent. In my new institution, this is unlikely to happen since I am the only aquatics user and the cage washer is already in use every day. Does anyone have suggestions or experience how to deal with this situation? I really don't want to have to resort to bleaching. Thanks for any help, Richard -- Please Note New Adress: Richard Kollmar, Ph.D. Visiting Associate Professor Dpt. of Cell Biology, BSB 3-65, Box 5 SUNY Downstate Medical Center 450 Clarkson Ave. Brooklyn, NY 11203 Tel. 718-221-6559 (office) Tel. 718-221-6563 (lab) FAX 718-270-3732 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090929/58e4bab5/attachment.html From yin.alessa from gmail.com Tue Sep 29 19:10:46 2009 From: yin.alessa from gmail.com (Chunyue) Date: Wed Sep 30 11:35:52 2009 Subject: [Zbrafish] protocol needed for phospho-smad staining Message-ID: <7ad948bb-3454-4c53-ab92-4dd1d74820bf@g23g2000yqh.googlegroups.com> Hi All, I tried to do immunohistochemistry using phospho-smad antibody (purchased from Cell Signal Technology). After several trials, I couldn't get any staining. Is anybody willing to share their protocols? BTW: I prefer to do the staining on whole-mount or vibrotome sections. Thanks a lot!