Nabble - Bio.net - Zbrafish

Bioprotocols online

2009-12-07T17:10:08Z

Sciclips has launched a unique database of bioprotocols for
researchers. Researchers will have free access to these bioprotocols
which have been extracted from published journal articles and patents/
patent applications. We are the first to list protocols from published
US patents, US patent and WO (PCT) applications. We have thousands of
protocols on molecular biology, proteomics, biological imaging,
virology, immunology, stem cells, drug Discovery, immunoassays, cell
biology, epigenetics, genomics, neurosciecne, plant biology,
microbiology, tissue microarrays and more. In addition to this, we
have also listed new sections on high throughput screening (HTS) and
biopharmaceutical assay protocols. All the protocols are linked to the
articles or patent/patent applications. All the protocols are linked
to the articles or patent/patent applications.. Please follow this
link to see the complete list of our bioprotocols online:
http://www.sciclips.com/sciclips/bio-protocols.do

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Most cited research reagents and kits

2009-12-02T23:51:30Z

An interactive graph showing most cited research reagents and kits
that are
currently available in the market. The data is based on recent
published journal articles, patents and patent applications. The
current research reagent/kits listings are: Transfection reagents, RT-
PCR, Real-time PCR, RNA purification, cAMP assays, Cytotoxicity assays
and Western blot reagents. Please visit:

http://www.sciclips.com/sciclips/most-cited-reagents.do?cat=TransfectionReagents\
&catId=1501

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Microinjection

2009-12-02T13:09:37Z

Dear ZFIN Members,
Our lab is in the process of increasing the number of micro injection
stations we have in our lab. Our current equipment is older and we
are looking for suggestions on the types of injectors and scopes
people are currently using as well as vendors who supply them. Any
input as to what is liked or disliked about the respective equipment
would also be appreciated.

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Re: plastic molds for microinjecting Zebrafish embryos

2009-11-26T03:19:27Z

On Nov 25, 9:27 am, Marina Raya <marina.r...@...> wrote:
> Hi all
> I would like to get some plastic molds for microinjecting Zebrafish
> embryos (as the ones described in the ZF book, chapter 5, cellular
> methods, A device to Hold Zebrafish Embryos during microinjection),
> but I can not find any provider.
> Do you know where can I order or get them?

Dear Marina

Please excuse the commercial response to this posting but this HAS
direct
relevance to your question and for others with the same technical
need.

A new methodology for this was published last week by the Kennedy lab,
Dublin.
Essentially, this uses a patented thermo-reversible hydrogel (CyGEL)
that
immobilizes the embryos, permits optimal positioning for injection and
then
simple recovery by briefly cooling, all done in a 3cm petri dish.

Alvarez Y, Astudillo O, Jensen L, Reynolds AL, Waghorne N, et al.
(2009)
Selective Inhibition of Retinal Angiogenesis by Targeting PI3 Kinase.
PLoS ONE 4(11): e7867. doi:10.1371/journal.pone.0007867

Refer to Fig. S1 for the text/images describing the technique.

For more information and ordering info on CyGEL:
http://www.biostatus.com/product/cygel/

Alternatively, contact me directly.

Best regards
Roy

Roy Edward
Biostatus Limited
Tel: +44 1509 558 163 Fax: +44 1509 651 061
roy@... www.biostatus.com
Company No: 3079239. Registered in England and Wales.
Registered office: Biostatus Ltd, 56 Charnwood Road,
Shepshed, Leicestershire, LE12 9NP, UK.

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Re: plastic molds for microinjecting Zebrafish embryos

2009-11-25T09:21:02Z

This company makes the molds that you are interested in, our lab has
purchased various models from them and they work very well:

http://www.adaptivesciencetools.com/

-Caitlin Stewart-Swift

Marina Raya wrote:

> Hi all
> I would like to get some plastic molds for microinjecting Zebrafish
> embryos (as the ones described in the ZF book, chapter 5, cellular
> methods, A device to Hold Zebrafish Embryos during microinjection),
> but I can not find any provider.
> Do you know where can I order or get them?
>
> _______________________________________________
> Zbrafish mailing list
> Zbrafish@...
> http://www.bio.net/biomail/listinfo/zbrafish

--
Caitlin Stewart-Swift
Senior Research Technician/Zebrafish Facility Supervisor
Department of Craniofacial and Molecular Genetics
Tel(617)636-4011
Caitlin.Stewart_swift@...

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plastic molds for microinjecting Zebrafish embryos

2009-11-25T01:27:23Z

Hi all
I would like to get some plastic molds for microinjecting Zebrafish
embryos (as the ones described in the ZF book, chapter 5, cellular
methods, A device to Hold Zebrafish Embryos during microinjection),
but I can not find any provider.
Do you know where can I order or get them?

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Alkaline Phosphatase staining of endothelial cells

2009-11-22T20:13:40Z

Hi there!
My lab has been trying to target the alkaline phosphatase staining for
the subintestinal basket. The protocol we follow is from Zebrafish:
Nusslein-Volhard and Dahm.My problem is that although the basket gets
stained, there's a lot of overstaining on the yolk, which does not
allow for proper imaging. I have tried adjusting the amount of NBT/
BCIP I add, but to no avail.
Could anyone tell me why the overstaining occurs, and how to prevent
it?
Thanks.

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cd41-gfp and rag2-gfp

2009-11-18T06:12:32Z

I am looking for vendors and care tips for these lines

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About embryo floating problem

2009-11-17T13:44:31Z

Sorry for the bothering!

After the overnight hybridization when the embryos are transferred from Hyb to 2*SSC the embryos just start floating on the buffer. I tried to drop some buffer using pipets to immerse them into the water but when you changed the buffer the embryos are just floating again.

Some of these embryos was not treated with proteinase K but they have the same problem. So I think it is not because of over digestion.

Does anybody have the same problem before? How could you avoid this from happening?

Thanks a lot for your help!

Journey.

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In situ hybridization services for hire?

2009-11-17T11:24:23Z

Hi, my lab just recently started experimenting with zebrafish as a
model system for dermatologic diseases and I was wondering if anyone
knew of any services/companies that do RNA in situ hybridization? If
not, does anyone sell pre-made reagents? We're in a bit of a time
crunch and while we already have a probe, we do not have the reagents
needed and it just makes more sense to commission someone who has
success with this procedure rather than us starting from scratch.

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Re: Red fluorescent reporters for transgenic fish

2009-11-04T10:51:12Z

Dear Wei,

Very likely your problem is due to the slow maturation of DsRed. mCherry should be better and is fairly bright. We have also had good success with TagRFP, which is extremely bright (available commercially from Evrogen). Another possibility is tdTomato, which is extremely bright. We have not used it ourselves but have heard good things from others. Don't know about maturation time.

There is an easy experiment to test whether maturation time is the issue: do a DsRed in situ on your transgenics, and see if the timecourse mimics the native gene.

best wishes,

Chi-Bin Chien

On Nov 2, 2009, at 5:19 PM, Wei Xia wrote:

Dear All,
We are wondering about what kinds of red fluorescent reporters are good for making transgenic zebrafish.
Our gene’s strongest expression occurs around 24hpf by in situ results. But when we used DsRed as the reporter we could not see any fluorescence until after 44-48hpf in our transgenic embryos. The fluorescence was pretty obvious at 24hpf if transient expression was checked by plasmid injection. So we guess the late detection of DsRed in these transgenic embryos is not because of the promoter we used is not complete or something. We thought this is might because of DsRed’s long maturation half time. We are thinking about using monomeric mCherry but worry about its low brightness compared to DsRed.
If you have any experiences of making RFP transgenic fish please share them with us!
Thanks in advance for your testimonies on this.
Wei
<ATT00001.txt>

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Trizol extraction - zebrafish embryos

2009-11-04T10:08:53Z

Has anyone encountered low 260/230 readings (<0.2) using the Trizol
RNA extraction. We're getting okay yields and great purity using 5
embryos (24hpf-72hpf)/sample, but are having problems with what we
assume is contamination.

We have already tried a number of things: bought all our chemicals
new, repeat chloroform extraction 2 times, do EtOH cleaning step 3
times. We homogenize manually with pestles.

Thanks for any advice.

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Zebrafish Husbandry Workshop at WAS 2010

2009-11-03T17:46:52Z

Flyer will also be posted on ZHA and Aquaneering websites and available via email by request. Spread the word!

6^th ANNUAL ZEBRAFISH WORKSHOP

San Diego Town & Country Convention Center March 2 – 3, 2010

The US Zebrafish Community has grown from isolated labs with wildly varying husbandry protocols and little institutional regulation to hundreds of Zebrafish Labs across the country, supported by multiple organizations including the Zebrafish International Resource Center (ZIRC), Zebrafish Information Network (ZFIN) and the Zebrafish Husbandry Association (ZHA). The move towards standardized practices continues as technicians, managers and veterinarians share information in forums such as this. Sometimes, however, it is easy to forget that

we are part of an even larger whole.

Our efforts support a research community that works to solve some of society’s most pressing problems, including cancer, genetic disorders, mental illness, heart disease, and environmental contamination. This research community spans national boundaries, and as the husbandry community grows in organization and maturity, we too will establish a dialogue with those practicing zebrafish husbandry around the world. We acknowledge that common interest and seek to promote an international dialogue in this program.

DDon’t Miss the ZHA’s Annual Meeting Monday evening 3/1/10 !! (time and location TBA)

TIME TITLE SPEAKER

Tuesday March 2, 2010

1:30 – 1:45 Introduction and Welcome Mark Francis/Aquaneering

Dan Castranova/ZHA

Zoltan Varga/ZIRC

1:45 – 2:10 Standardization of Zebrafish Nutrition – Preliminary Steve Watts/University of Alabama

Summary and Recommendations

2:10 – 2:35 Environmental and Genetic Determinants of Sex in Christian Lawrence/Children’s Hospital Boston

Laboratory Zebrafish

2:35 – 3:00 Medaka Husbandry Review for ZF Professionals Kenji Murata/University of California, Davis

3:00 – 3:30 BREAK

SPECIAL SECTION: Peer Submitted and Reviewed Presentations Coordinated by ZHA

3:30 – 3:50 Zebrafish Husbandry - Maintaining and Rearing Albino Kay Stewart/University of Notre Dame

Zebrafish

3:50 – 4:10 Living Together Before Mating: Reproductive Success in Sarah Doty & David Shanholtzer/Stowers Institute

Single Sex and Mixed Sex Tanks in Zebrafish (Danio rerio) for Medical Research

4:10 – 4:30 Automated Feed System- Advanced Husbandry Chris Obenschain/Aquatic Solutions-Tecniplast

Technologies

4:30 – 4:50 The Effect of Stocking Density on Embryo Production of Eric Herbst/Aquatic Habitats

Zebrafish, Danio rerio, Housed in the Mini Mass Embryo

Production System, miniMEPS™

4:50 – 5:10 A Novel Group Spawning Chamber for Zebrafish Isaac Adatto/Children’s Hospital Boston

5:10 – 5:30 A New Method for Rearing Larval Zebrafish with Type Jason Best/Children’s Hospital Boston

"L" Saltwater Rotifers (Brachionus plicatilis)

Wednesday March 3, 2010

8:30 – 8:50 A Retrospective Study of the Occurrence, Presentation R. Smolowitz^/Roger Williams University

And Microscopic Appearance of Abdominal Tumors in

Zebrafish held in a Laboratory Animal Facility

8:50 – 9:10 Ultrasound Enhanced Transport of Methanol into Amrit N. Bart/Asian Institute of Technology,

Zebrafish Danio rerio Embryo Thailand

9:10 – 9:30 Development of Protocols for Zebrafish Sperm Cryo- Huiping Yang/Aquaculture Research Station,

preservation in Different Packaging Containers Louisiana State University Agricultural Center

SPECIAL SECTION: Fish Diseases Coordinated by Michael Kent

9:40 – 10:00 Morphological Effect of the Stress Response in Fish Claudia Harper/Amgen

10:00 – 10:30 BREAK

10:30 – 10:50 Pleistophora Hyphessobriconis (Microsporidia), the Michael Kent/Oregon State University

Cause of Neon Tetra Disease, in Zebrafish Research Labs

10:50 – 11:10 Update on an Improved PCR Test for Pseudoloma Justin Sanders/Oregon State University

neurophilia, and its Application for Non-lethal Testing

11:10 – 11:30 Zebrafish Health and Husbandry at U Washington George Sanders/University of Washington

11:30 – 11:50 Review of Several Interesting Diagnostic Cases Trace Peterson/University of Oregon

11:50 – 12:10 Natural Transmission of Mycobacteria in Fish Don Ennis/University of Louisiana

12:10 – 12:30 Zebrafish Health Issues at UC San Diego Keith Jenne/UCSD

12:30 – 1:30 LUNCH

1:30 – 2:00 KeyNote Speaker

The Study of Social Control of the Brain Dr. Russell Fernald/Stanford University

2:00 – 2:25 Zebrafish Core Facility and ZigZag Forum at the Mayo Gary Moulder/Mayo Clinic

Clinic: Influencing Research through a Synergy Model

2:30 – 2:50 British Legislation and Husbandry Standardization Carole Wilson/University College London

2:50 – 3:00 BREAK

3:00 – 3:30 A Global Update on Guidelines for Fish Research Gilly Griffin/Canadian Council on Animal Care

3:30 – 4:00 Summary: Guide for the Care and Use of Laboratory Monte Matthews/University of Oregon

Animals – Changes that Affect Zebrafish Husbandry

4:00 – 5:15 The Big Picture: Regulation, Standardization, International PANEL DISCUSSION

Recognition & Cooperation

5:30 – 7:30 TOWN HALL MEETING

FOR MORE INFORMATION:

WORKSHOP: bobbi@...

ZHA: admin@...

Conference Details and Registration: http://www.was.org

(Note: ZHA Members receive discounted registration!)

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RE: zebrafish gill disease?

2009-11-03T09:59:19Z

Our conductivity is routinely between 500-700 and we don’t have an issue with this.

Hope this helps,

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@...

Admin Assistant: Anna Schmedel (609) 258-5028

From: zbrafish-bounces@... [mailto:zbrafish-bounces@...] On Behalf Of Evert-Jan van den Brandhof
Sent: Tuesday, November 03, 2009 1:49 AM
To: zbrafish@...
Subject: [Zbrafish] zebrafish gill disease?

Hello,

Is anyone familiar with (what looks as) fast growing epthelium on upper gill covers?
We do'nt have any problem with egg production but it may cause some stress on breathing capacity?

see picture:

could anyone reconfirm this could be caused by conductivity?

system: Zebtec (http://www.tecniplast.it/_assets/products/CAT012_ING_Internet.PDF)

pH= 7.5
Conductivity= 500 µS
Temperature= 27,5 °C

Conductivity and pH are automatically adjusted with Seesalt and Sodiumbicarbonate.

Thanks in advance,

Evert-Jan

¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤
Evert-Jan van den Brandhof, BSc
National Institute for Public Health and Environment (RIVM)
Laboratory for Ecological Risk Assessment (LER)
P.O. box 1, 3720 BA Bilthoven, The Netherlands
Tel: (+31)30-2743544
http://www.rivm.nl/milieuportaal/
¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤

Disclaimer RIVM

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zebrafish gill disease?

2009-11-02T22:48:44Z

kieuw effect.jpg (295K) Download Attachment

Red fluorescent reporters for transgenic fish

2009-11-02T16:19:08Z

Dear All,

We are wondering about what kinds of red fluorescent reporters are good for making transgenic zebrafish.

Our gene’s strongest expression occurs around 24hpf by in situ results. But when we used DsRed as the reporter we could not see any fluorescence until after 44-48hpf in our transgenic embryos. The fluorescence was pretty obvious at 24hpf if transient expression was checked by plasmid injection. So we guess the late detection of DsRed in these transgenic embryos is not because of the promoter we used is not complete or something. We thought this is might because of DsRed’s long maturation half time. We are thinking about using monomeric mCherry but worry about its low brightness compared to DsRed.

If you have any experiences of making RFP transgenic fish please share them with us!

Thanks in advance for your testimonies on this.

Wei

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zebrafish injections

2009-10-29T07:08:48Z

dear all,

we are planning to purchase a new microinjection set-up apparatus for our lab.
we currently have femtojet micromanipulator 5171 from eppendorf. and the
microscope is from zeiss; stemi 2000.

i am on the look-out for a manipulator which can make an axial movement; is
injectman good? and additionally, i appreciate your input of a microscope which
has light source from below.

your suggestions will be of great help.

thank you very much,
sanjita

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Biosecurity Measures

2009-10-23T10:44:54Z

We have a policy the restricts entry into our facility by anyone who has been in another fish facility or had contact with fish from another facility within the last 24 hours. The idea is that people should have at least taken a shower and changed clothes before entering our facility. This policy was created to prevent the introduction of any pathogens that could be transported on a persons skin and clothing. I would like to know if other zebrafish facilities have a similar policy? If so, were there any specific reasons why this policy was implemented?

Best Fishes,

Jim Burris

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Looking for groups that use zebrafish

2009-10-16T08:30:41Z

Good day, I am looking for a listing of as many (all) research groups
that use zebrafish in their model. Does anybody have a suggestion on
where I could find information like that? It would be much easier than
randomly searching through all the groups I can find using a generic
search. Very much appreciated, Scott.

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in situ robots

2009-10-06T17:39:22Z

Hi All,

i am currently looking at the idea of getting an in situ robot.

i was just wondering if people who have used either the BioLane HTI,
or the InsituPro VSI could send me some thoughts on whether you are
happy with the machine, or think there is a better alternative out
there?

many thanks,

jason

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Cage washers and detergents

2009-09-29T18:38:56Z

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

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protocol needed for phospho-smad staining

2009-09-29T17:10:46Z

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!

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Re: On raising Zebrafish larvae

2009-09-25T08:34:38Z

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

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post anesthesia procedure

2009-09-25T00:05:16Z

Hi all,

We just let them recover in a 8L (1.66 gallon) tank, in fish water. After a few minutes, when they swim well, we put them back in the system.

Hope it helps,

Marie Winandy
GIGA Zebrafish Unit
Avenue de l'Hopital, 1 / B34
4000 Liege-Belgium

[mwinandy.vcf]

begin:vcard fn:Marie Winandy PhD n:Winandy PhD;Marie org:Universite de Liege;GIGA Zebrafish Unit adr:;;Avenue de l'Hopital 1/ B34;Liege;;4000;Belgium email;internet:mwinandy@... title:Zebrafish Platform Manager tel;work:+32.4.366.99.71 tel;fax:+32.4.366.41.98 tel;cell:+32.476.97.25.33 url:http://www.giga.ulg.ac.be/jcms/c_5418/giga-animalerie-et-transgenese-zebrafish version:2.1 end:vcard

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Post-anesthesia procedures for zebrafish

2009-09-24T09:33:28Z

What are other facilities’ post-anesthesia procedures for zebrafish?
Why are these procedures utilized?

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EvoDevo Job at UMass Amherst

2009-09-23T12:35:51Z

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@....

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@...

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Reactome Pathway Database User Survey

2009-09-23T12:18:31Z

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

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Morpholino oligos injection

2009-09-23T07:28:58Z

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

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GFP staining adult wax sections

2009-09-15T01:42:40Z

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

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breeding transgenic zebrafish

2009-09-14T06:41:03Z

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

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embedding embryos in plastic

2009-09-11T11:42:04Z

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

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Re: On raising Zebrafish larvae

2009-09-10T22:55:58Z

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

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RE: On raising Zebrafish larvae

2009-09-09T08:52:30Z

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@...
Admin Assistant: Anna Schmedel (609) 258-5028

> -----Original Message-----
> From: zbrafish-bounces@... [mailto:zbrafish-
> bounces@...] On Behalf Of Lawrence, Christian
> Sent: Tuesday, September 08, 2009 10:51 PM
> To: chubbyjayu; bionet-organisms-zebrafish@...
> 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" <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
>
> _______________________________________________
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>
>
> 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.
>
>
>
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Re: On raising Zebrafish larvae

2009-09-09T06:29:02Z

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

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Re: On raising Zebrafish larvae

2009-09-08T19:50:41Z

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" <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

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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.

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