Ed, I am sure that with your superior photography skills you would be more than happy to grace this forum with your morphing pictures, they are so adorable from eggs to tads. We at fatfrogs have asked Ed on numerous occasios to post these pictures as do the other breeders on that forum but he has never obliged us. Ed never spoke to me about the sex of the frog after I recieved her. My only communications with Ed have been via pm and email. I would like you to post proof of this statement Ed. The sex of the frog was determined by my vet here who during the initial necropsy found it unremarkable and recommended pathology. Ed refused to respond to any of my emails or pm's after telling me that he honored his guarantee and would not offer a refund, I posted that response previously but if anyone here would like me to do so again I will. Ed if you would please post the previous pm's that you recieved from me, the ones that I posted your responses to, this would help to clarify the situation. Especially for those who do not believe that I "did my research". I asked alot of questions these frogs, hence the responses that I previously posted.
I won't go quote by quote and take up extra space so I jotted some notes to answer the above questions to the best of my ability, if I have missed one please let me know and I will do so ASAP.
Seamus, you were likely asked if your head was going to explode as a compliment by Jamie, she has never insulted anyone on any of the other forums that we are members on. Please take it as such, she recognizes your level of intelligence as have we all who have read your posts on this thread.
The cost of the frog was 30.00 and the shipping that I paid was 55.00 for a total of 85.00. I explained the need of the necropsy in a previous post. I needed to verify that I did not cause this frog to die. My vet determined the necessity of pathology and I agreed. I needed a clear conscience as I just fell in love with her on sight. I had waited 2 months to get her. As I stated there are no blue phase whites in my area.
There are only 2 places that test for Chytrid Fungus that I found on google and in speaking to Mr. Wood decided that I would use pisces-molecular. He is of enormous comfort and assistance. Their lab is very busy and the results can take 2-4 weeks. I will post the email that he sent to me as it is very informative.
My local vet has never heard of chytrid fungus and I called every one from herp vet connection from here to Richmond and even spoke to the state vet here because I know her personally. She has not heard of it either.
I did ask if these frogs could fall because I have never owned a whites before. I know that they are tree frogs and are able to jump and therefore need a large enclosure which is why I purchased the largest one that I could find. I was very doubtful that she had fallen and was only concerned because of her lethargy. I did attempt to feed her. The morning of her first day here I did open the door to the enclosure while ducked down since I knew she was behind the rockpile. I have long steel tongs and slowly edged my hand up where she could not see me or my hand and moved the cricket back and forth where I knew she could see it. I did not and would never poke, prod or forcefeed an animal, unless it was required to save their life but never to a new frog that didn't "know" me much less have time to acclimate.
I did not state that the frog had bloodshot eyes and neither did Lars. He stated that her sticky toe pads appeared bloodshot.
My horned fogs were treated for CF when I got them. The cranwell was my first frog and he, definitely a male because he calls and is in breeding season so his nuptual pads are quite prominent at the present time. I joined fatfrogs to learn the proper care of these animals since care sheet after care sheet said that a semi aquatic tank and gravel were safe. I use coco fiber only. I removed him from his enclosure and sterilized it and began treating for CF as soon as I read the sticky thread on fatfrogs made by Steven Busch. My ornate female was purchased locally as well and she started treatment the day that I got her.
The treatment for CF is lamasil at spray,not lotion. 10 pumps into 200ml of dechlorinated water. 5 minute soaks for 10 days. After each soak the breeder boxes that were their temporary homes had to be sterilized before placing them back into them. The enviornment can reinfect them and must be sterilized between each treatment. The treatment causes shed and revealed beautiful patterns and a marked color enhancement on my ornate. Their appetites improved and they are both currently doing very well.
I was using the terrarrium that I purchased for the whites with paper towels to keep it as clean as possible, monitor droppings and not to move her around too much. It was large enough so that she had room to move and plenty of places to hide and feel safe.
It was not that bright an enviornment unless she was used to a nearly dark one. I have not seen Ed's enclosure where she was so I do not have that as comparison. My bedroom faces the front of my house and with no trees can get bright which is the reason that I keep my blinds closed and I have purple sheers over them during the day. I live with my daughter and of course have concerns about unwelcomed guests or burglars so we keep the blinds closed throughtout my home all the time.
I had 2 25 watt nightglo bulbs on the canopy top for the terrrarrium. They do not produce much light at all. It is obviously lit when during the day but the enclosure was not in front of a window. It recieved indirect sunlight only.
I took pictures of the necropsy today and hope that they are legible.
Some of them were quite blurry but these were the best ones that I could get. I really wish I owned a scanner.
This is the email sent by Mr. Wood at pisces-molecular, I cannot open the links at home, I don't have the best computer but my friend has a better one, no scanner though, and I opened the links there, I hope the members here are able to open them.
Flag this messageRE: chytrid fungus testingSaturday, August 16, 2008 4:23 PM
From: "John Wood" <
[email protected]>Add sender to Contacts To: "'kelly brown'" <
[email protected]>Message contains attachmentsPCR_Collection_Protocol_2004.doc (857KB), General Instructions for Collecting 1-1-08.doc (30KB)Dear Kelly,
We would be pleased to PCR test an amphibian sample for Batrachochytrium dendrobaditis for you.
Attached are two files which answer most of the common questions about collecting and preserving samples for B.d. PCR testing. The first is a protocol for collecting amphibian toe clip, skin scrape or skin swab samples for PCR testing from Lauren Livo whom we work with at the Colorado Division of Wildlife. The second file from Pisces continues on from Lauren’s with instructions and information about packaging and sending samples for B.d PCR to us.
Several additional notes beyond what is covered in these two files:
Because our experience with the 2 ml screw-capped microfuge tubes with a captive O-ring gasket recommended in the instruction files has been very positive - nearly zero leaking tubes in thousands of samples - we strongly recommend using these tubes for your samples. In response to requests from many customers who don't want to buy 500 vials at a time, or don't want to spend the time to be sure they're getting the right tubes or ethanol, we are now offering "swab sample collection kits" consisting of 50 2ml screw-cap tubes filled with 1 ml of 70% ethanol; 50 swabs; ziploc bags for everything; and a box that can be used for return shipping to Pisces. The cost of the 50-sample kit is $50 plus shipping.
The "general instruction ..." file specifies the materials for pooling samples, but doesn't yet explain the two alternative sample pooling strategies we have developed and offer. We can pool up to 8 swab samples into one DNA prep+PCR sample. Either you can do the pooling, or we can do it here. Although there is an extra charge if we do the pooling (see below), we generally recommend that new customers let us do it, at least for the first pass of sampling at new sites where nothing is known about the prevalence or severity of B.d. The chytrid PCR assay can detect less than 1 zoospore, so it is extremely sensitive to sample cross-contamination; we have procedures in place to pool samples efficiently without cross-contamination. Also, if we do the pooling, we can pool 1/2 (by volume) of each individual sample. Then if a given pooled sample is PCR positive, we can go back and prep and PCR test the remaining half samples individually. This works out well when one wants to know a true B.d. incidence level without spending lots of money on a large batch of samples that all turn out to be PCR negative.
We do not yet require or have specific sample submission forms; all we require are: 1) The name of the sample submitter; 2) A hard copy list of the number of samples submitted and the individual sample IDs; and 3) To whom the report and the invoice should be sent.
The current price for carrying out the DNA extraction and the chytrid PCR test for samples sent to us is a $20 accession fee per group of samples plus US$27 per sample. For samples that you send as individual samples and we do the pooling, there is additional $35 pooling charge for each group pooled (for example, 8 individual samples pooled into one DNA prep + one PCR test would cost:$35+$27 = $62. For samples sent to us already pooled, the charge is $37/pooled sample. Samples sent in tubes or vials other than those recommended are subject to a 20% surcharge. Samples which leak in transit – and therefore require extensive additional anti-cross contamination measures at Pisces - will be subject to a $250 per batch leak surcharge. If we discover a leak in a sample batch, we will immediately email or call you to report the leak and allow you to decide whether we should proceed, or let you resample and submit another (hopefully) non-leaky batch of samples. Individual swab sample collection kits are $50 per 50 samples, plus shipping. Kits for customer pooled samples (50 ml tubes with 70% ethanol and 8 swabs per tube, instructions) are $80 for 10 pooled samples.
Our terms are: Payment is due upon receipt of the sample test results report. All we require from new customers is a name and address to send the invoice to, and a P.O. number if required by the customer. Unfortunately, we are not able to accept credit card payments at this time. Samples should be packed per the instructions in the "General instructions" and shipped FedEx, DHL or UPS; 2nd day delivery is fine.
Our normal turn-around time from receipt is two to four weeks. However during the summer field season, or in the fall when all the field season samples have been sent in, our turn-around can slow to four to six weeks (our current turn-around time as of 8/16/08 for normal samples is ~4 weeks). If you have a need for faster results, for animals in quarantine or a report deadline, please call us. Although we haven’t completely formalized our rush sample times and charges, we can very likely work something out that meets your needs – our fastest sample turnaround time to date was 9 hours (FedEx Priority delivery of the samples in the morning; PCR results emailed by 6 PM!) (currently we have been quoting < 1 week turnaround for $54/sample and < 48 hr turnaround for $108/sample). We will email and/or send out a signed hard copy of the test results report, whichever is needed by the sample submitter. Test results are kept confidential and not shared with anyone other than the sample submitter or their organization without express permission from the submitter.
(Kelly, since your tree frog has already died, a toe clip sample will work better than a belly skin swab. The file from Lauren Livo does explain how to collect a toe clip. You will need to preserve it in ethanol. If you can't get any lab grade 70% or 95% ethanol, "Everclear" from a liquor store will work; it is 95% alcohol. Please make sure that you do not dilute the alcohol you use with tap water that contains chlorine; chlorine will destroy the DNA in the sample!)
Let me know if I've missed anything important or you have any more questions. We look forward to being of service to you.
Thank you,
John Wood
Pisces Molecular
2200 Central Avenue, Suite F
Boulder, CO 80301
USA
v: 303-546-9300; f: 303-546-9400
I hope that this is sufficient to answer the questions that everyone here has asked of me, again if I have not please ask again.
Ed, please take the time to read this as well as to answer the questions that I have asked you in the previous threads.
