:Correction to post--calculations to prepare 10% formalin solution from 37% stoc

[JCW1925 at aol.com]

RE:Correction to post--calculations to prepare 10% formalin solution from 37% 
stock

Dear Breeders, Monarch Watchers, Lepidoptereist:

 The following attached  8/20/99 post to the Farmers list contains a gross 
calculation error that may lead to ineffective sterilization of monarch eggs 
and, or inaccurate data as to the effectiveness of  surface decontamination 
of monarch eggs with formalin. The final concentration of the formalin that 
was posted on the list(s) came to 0.962% not the intended 10%.  (2.6 ml  of 
37% stock/brought to 100 ml with water = 0.962% solution)  The correct 
calculation is  X ml  37% / 100ml = 10%.  X = 27.02 mL.  Therefore, to make 
100 ml of a 10% solution of formalin you need to add 27 ml  of 37% formalin 
(stock)  and 73 ml of water.  In my lab, we use modified ( conatining 
biological detergents) formalin solutions with heat to denature cells on 
slides so we can target individual chromosomes with apha-satelite probes. 
(FISH)   A  properly prepared 10% solution of formalin should be effective in 
 denaturing ( modifying the tertiary and secondary stuctures) of  the protein 
coats of viruses that may be attached to monarch eggs.  We have just recently 
started experiments using a 10% solution of buffered histological grade 
formalin, pH 7.1)  to determine whether or not a 10% solution of formalin is 
is effective in killing the spores of the protozoans Ophryocystis and Nosema, 
 the length of time necessary to effectively sterilize monarch eggs and how 
long the monarch eggs can tolerate 10% buffered formalin without any 
detrimental effects.

    Should any one need  10%  buffered formalin, we have this available in 
30ml jars that can easily be shipped  under IATA limited quantities regs.  We 
should have this new product soon listed on our web site at 
www.educationalscience.com.  We have formulated an artificial monarch diet 
that eliminates the problems (vector for Oe and other  larval diseases) 
associated with  feeding monarch caterpillars milkweed.  The diet works well 
and 5th instar larvae do not require supplemental milkweed.  If the 10% 
formalin method is effective in sterilizing monarch eggs and   killing the 
spores of the protozoans Ophryocystis and Nosema,  we should be able to 
provide sterile monarch cultures, as our artificial diets are gamma 
irradiated.  I am not sure that this is a good idea, as I believe that 
natural diseases are natures intended way of controlling monarch populations.

  For more information on our  sterile artificial monarch diet,  please visit 
our web site at www.educationalscience.com.  We also provide Monarch larvae 
(cultures) year round,  rearing and breeding kits,  pesticide-free milkweed 
plants, milkweed seeds by the pound, as well as butterfly farming and  insect 
rearing supplies, tagging cages,  breeding cages, entomology kits and 
equipment, butterfly nets, and biotechnology education kits and equipment.

  We offer as a free service to our customers who have purchased livestock 
free screening for  Oe  spores.  Should any one in Texas require this service 
for free, please contact us at 1-800-299-6791 or at www.educationalscience.com

James Wiggins
Educational Science
www.educationalscience.com
Toll free: 1-800-299-6791



   Two chemicals are commonly used to "sterilize the surfaces of insect,
particularly Lepidoptera, eggs; chlorox and diluted formaldehyde (formalin).

T
The formalin method:

Formalin is a diluted form of formaldehyde. The concentrations used to
sterilize eggs range from 6-10%. Formaldehyde is nasty stuff, a potential
carcinogen, and should only be handled in well ventilated areas. If you
purchase formaldehyde, you will receive a stock solution which is 37%
formaldehyde. This needs to be diluted as follows: To make a 6.5% solution
for sterilization of eggs, add 2.6 mls of formaldehyde to 100 mls of
distilled water. For convenience, and to avoid handling the formaldehyde
bottle frequently, you might make up a stock solution of several liters of
formalin.  It won't spoil. Once you have made up the stock solution, you
can proceed with egg sterilization following the same steps as outlined for
chlorox above.

Formalin sterilization of egg surfaces works well for bacteria, viruses,
molds,  and fungi. We do not know whether this method is effective in
killing the spores of the protozoans Ophryocystis and Nosema.

he chlorox method:
>
>1.  In a 500ml. beaker, mix the following thoroughly: 10ml. Clorox ,
>400ml. of distilled water and to this add a 1/4 drop of dish soap.
>2.  Place leaves with eggs in a net bag after trimming excess leaves away.
>Immerse the leaves and eggs for 6 minutes.
>3.  Move the leaves/eggs to 500ml. of distilled water for 10 min. to rinse
>off the excess Clorox.
>4.  Air dry the leaves/eggs.
>6.  Place leaves/eggs into 1/4 pint container (with a clear plastic lid)
>lined with damp filter paper or paper towel.
>7.  Newly emerged larvae will crawl to the top of the container.
>8.  Use a wet paint brush to move larvae onto hostplants. (The host plants
>can be either cut or living).

This method works for  bacteria and viruses but is not as effective in
killing the spores of the protozoans, Ophryocystis and Nosema.


If you use this method, please let us know whether it is effective or not.
For a valid test, you need to establish that the females used to produce
the eggs and/or the eggs themselves are infested with spores. Please see
the web site for information on how to detect the spores of Ophryocystis.

Chip

------------------------
Monarch Watch
Email:  monarch at ukans.edu
WWW: http://www.MonarchWatch.org/
Dplex-L:  send message "info Dplex-L" to Listproc at ukans.edu
Phone:  1 (888) TAGGING (toll-free!) -or- 1 (785) 864 4441
Fax:  1 (785) 864 4441 -or- 1 (785) 864 5321
Snail:  c/o O.R. Taylor, Dept. of Entomology, 7005 Haworth Hall, Univ. of
KS, Lawrence KS 66045


The formalin method:

Formalin is a diluted form of formaldehyde. The concentrations used to
sterilize eggs range from 6-10%. Formaldehyde is nasty stuff, a potential
carcinogen, and should only be handled in well ventilated areas. If you
purchase formaldehyde, you will receive a stock solution which is 37%
formaldehyde. This needs to be diluted as follows: To make a 6.5% solution
for sterilization of eggs, add 2.6 mls of formaldehyde to 100 mls of
distilled water. For convenience, and to avoid handling the formaldehyde
bottle frequently, you might make up a stock solution of several liters of
formalin.  It won't spoil. Once you have made up the stock solution, you
can proceed with egg sterilization following the same steps as outlined for
chlorox above.

Formalin sterilization of egg surfaces works well for bacteria, viruses,
molds,  and fungi. We do not know whether this method is effective in
killing the spores of the protozoans Ophryocystis and Nosema.

If you use this method, please let us know whether it is effective or not.
For a valid test, you need to establish that the females used to produce
the eggs and/or the eggs themselves are infested with spores. Please see
the web site for information on how to detect the spores of Ophryocystis.

Chip

------------------------
Monarch Watch
Email:  monarch at ukans.edu
WWW: http://www.MonarchWatch.org/
Dplex-L:  send message "info Dplex-L" to Listproc at ukans.edu
Phone:  1 (888) TAGGING (toll-free!) -or- 1 (785) 864 4441
Fax:  1 (785) 864 4441 -or- 1 (785) 864 5321
Snail:  c/o O.R. Taylor, Dept. of Entomology, 7005 Haworth Hall, Univ. of
KS, Lawrence KS 66045