# Ask Biotracking.com



## NubianSoaps.com

Members of DGI. Please leave your questions for Chuck here. These are questions for Chuck to answer, not members of the forum, please lets keep the chatter to a minimum on here. Vicki with her owner hat on!


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## MF-Alpines

Chuck:

On one of the threads, Rose explained the difference between AGID and ELISA. But I still have a couple of questions:

But I don't understand this part: Each lab could potentially use different CAE virus proteins, thus explaining some of the differences in ELISA titers between labs on the same goats.

I don't understand the "...different CAE virus proteins..."

And why can/does Biotracking give titer levels and others, even using ELISA, cannot?

Thanks!


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## ChuckAtBioTracking

Hi Cindy - It probably doesn't make a lot of difference which protein is used in developing a test - there really are not that many proteins to choose from and for the most part the best target is the capsid or coat protein. Even if there were widely different coat proteins, the response of the goat exposed to the virus is to make what are known as polyclonal antibodies, so essentially the animal will be making different antibodies to as many different viral proteins the immune system "sees".

The test kit we use has three components; the sample plate, a specially modified, or "testing" antibody against CAE, and a colorless substrate which can turn blue in the presence of the specially modified antibody. In our assay, the test plates (think a plastic plate about the size of a stack of 3 x 5 index cards with 96 wells) are coated with viral protein which captures any CAEV antibody in the serum. If there is no antibody in the serum sample then nothing is captured, and the coating protein remains 'naked'. We wash out the sample, then add the "testing" antibody. If there was CAE antibody in the sample, it will partially or totally (depending on how much was in the serum) block the binding of the "testing" antibody. If there is no antibody in the sample, then the "testing" antibody binds at a 100% level. The colorless substrate is added and if the "testing" antibody is present, the well will turn blue. Depending on how much antibody is in the sample, it will compete out and allow only some of the "testing" antibody to bind, and the resulting blue color will be less intense. So the more antibody in the sample, the less "testing" antibody will bind, with a resulting lighter blue color (this is the "inhibition" concept). We can measure the difference between the dark blue 100% binding in the negative control, and the lighter blue color in the positive control, use a little algebra to determine the cutoff values of the controls and the numerical value of each of the samples, and compare them to the cutoff. In other words, since antibody in the sample "inhibits" binding of the "testing" antibody, the blue color is reduced and we can calculate the percent inhibition which we report to you. It's not truly an antibody titer, but it is related to the amount of antibody in the serum. 

A lot of info there so I hope it's not too confusing. As my granddaughter says "blah blah blah blah science"

Chuck


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## ChuckAtBioTracking

OK - here is the "official" protocol for the days post breeding study. I've included a milk aspect to it as well so if you would like to do both blood and milk, let me know.

Chuck

BioTracking LLC REQUEST FOR ASSISTANCE:
Study GS2: POSTBREEDING GOAT BLOOD/ MILK SAMPLING

Purpose: BioTracking LLC is requesting your assistance in collecting blood (and milk) samples from goats starting at 20 days post breeding.

BioTracking will provide or reimburse for milk containers, tubes, needles, needle holders and Federal Express shipping labels. Please submit a request of the material you will need before the start of the project to allow time to process. 

Timeline: The project is scheduled to be started as soon as possible. 

Project overview:
1.	Use dairy or meat goats.
2.	Either AI or pen- or pasture-bred does, with no contact with a buck for the duration of the study.
3.	We would like as many does as possible, up to 300 - 400 different goats in total.
4.	Begin sampling does when they are 20 days post breeding. Continue to collect samples every two days through day 36, then again (if scored as Pregnant through day 36) at days 45, 60 and 90 DPB. 
5.	Not all goats need to be collected at the same time, but ultimately the 12 blood samples will be needed from every goat included in the study.
6.	Collect as much serum as possible (greater than 5 ml of whole blood). 
7.	For the milk study, collect milk samples as pre-milking strip. Collect samples in DHI containers with preservative.
8.	Record goat ID’s so that duplication can be avoided at time of sample collection and/ or data processing. Also record farm name on tube if sampling occurs at more than one farm. 
9.	Exact data for each goat needs to be recorded at time of sampling. Necessary information includes goat ID, DPB at time of sampling, farm name if collected on more than one farm and breed of the goat.
10. Store samples in the refrigerator until they are sent to BioTracking. If possible, samples should be held until the day 36 draw, then sent to BioTracking. The remaining 45, 60 and 90 day samples should be sent to BioTracking after that DPB draw. If the goats for each time point are drawn on different days, the samples should be held until all samples for that particular day have been collected. Carefully package samples. Make sure each tube is clearly identified and appropriate cow data is included in the package or sent electronically. Make sure blood (and milk) samples are in a leak proof container.
11.	Email BioTracking with the size and weight of the package so we can prepare and send you a FedEx shipping label.


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## buckrun

Thanks for that. Is the same process used to test colostrum samples?
Lee


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## Rosesgoats

Hi, Thanks for the explanation. What do you think would contribute to different labs having different results when both using ELISA? Is it more a difference in protocols and procedures between labs or more of a difference in the blood samples themselves? Can two blood samples from the same goat come up with different results?


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## Qvrfullmidwife

Regarding CAE testing--how common is it to see a CAE test result of "0"?


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## ChuckAtBioTracking

It's pretty common to see a 0 on the CAE assay. The way the standards are set up to do the inhibition calculations, the "negative" is not the absolute minimum value, unlike the positive value which is 100.000% So because the negative control is somewhat "positive" it is possible for goats to be below the value of the negative control. In fact, most of the 0 values are negative numbers, but I decided to call 0 the minimum value I would report, jsut to prevent confusion from people getting a report that says Gracie -16.54% Not sure if I succeeded on the non-confusion issue or not! 

For the colostrum sample, I've tried using milk and colostrum for detecting CAE. It works kinda sorta on milk, but colostrum is just awful to work with, and milk isn't that much better really. But yes, as far as the assay is concerned, when I was playing around with milk and colostrum as a possible test sample I treated it just like I would serum. It's just too messy to work with, and the USDA license so far is good only for blood.

What I'm doing in the days post breeding study is looking at both blood and milk as a pregnancy testing sample. It's still kind of messy as a source of sample and not quite a sensitive as the blood, but has some advantages for the producer. I'm curious how milk would work for the goat world, if goats are treated as cows are in the milk producing world (see my thread "stupid lab-rat question")


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## swgoats

Have you kept any statistics as to how many goats test positive for CAE?


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## ChuckAtBioTracking

Yes I have all of the reports back to when we first started running CAE (started in Jan 2008) so we could do some data mining and see what kinds of numbers we get overall. I would suspect overall in North America the infection rate could be as high as 75% since there are a lot of breeders who figure it's so hard to eradicate and most animals are asymptomatic that there really isn't that big a need for prevention. Sometimes education helps on this, sometimes it's a case of don't confuse me with facts my mind is made up. I'm happy to help any way I can in the education department.

Chuck


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## Faithful Crown Nubians

Why do the red topped blood tubes have an expiration date? Can they still be used for pregnancy testing? (for goats) Thanks!


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## ChuckAtBioTracking

Some red top tubes have a clotting additive in them, many tubes have nothing but the vacuum. I think the "expiration" date has more to do with how long you can expect the tube to hold the vacuum. As with most expiration dates, they are pretty conservative so don't feel you have to toss out the expired tubes. Kind of like the labels on yougurt - I keep eating them well past the date . . .

For the tubes, you'll know soon enough if they are "bad" - they won't fill for you. 

Chuck


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## ChuckAtBioTracking

Hello all - I just sent a rather lengthy reply to a client who has some real frustrations (and rightly so!) with past and current results of the CAE testing. I thought I'd go ahead and post it here in hope it may answer some questions others of you may have. I'm sure I covered the same things earlier, and there may be some new things here. Hopefully there are no contradictory things! I'm sure someone will point any out to me!

Greetings - well there are a lot of unanswered questions about CAE simply because the virus is so versatile in how it infects, how it shows symptoms (or doesn't in a large majority of the cases) and when it becomes active after an infective event.

In the case of the goat in question, I wouldn't become too concerned about the inhibition numbers drifting from 0 to 10 or so. Several things can affect that, from random drift in the assay (this is true for any clinical assay) to a different manufacturing lot from one test to the next. I don't get concerned about the value of the numbers unless they start to push up into the 25+% range. Then I suggest testing another sample in 4 - 6 weeks to see if something in the serum may have been giving a bit of a false positive lean to the results.

The risk one runs with maintaining a mixed herd is that despite all the proactive things one does, CAE is on the farm and the potential for it to spread is very real. It is true that probably 90% of the infectious events are due to consuming contaminated colostrum and/or milk. But this leaves an additional 10+% for other routes of infection. Anything containing white blood cells can harbor the virus: blood, milk, lymph, nasal mucosa, saliva, pus from infected wounds etc. So any contact with these fluids (licking, biting, rubbing open wounds together (as in head-butting), drooling or dripping into common feed bows and water troughs can lead to a possible infectious event. The virus isn't considered to be transmitted as an airborne event, however it is possible that an animal with a high virus titer and a very runny nose could sneeze in the face of another animal with a resulting infection.

These are probably not too common, but nonetheless are potential modes of viral transmission. But to avoid becoming too paranoid about things, the virus is quite fragile and does not do well for very long outside of the host. Also, as with many if not all viral or bacterial infections, the viral or bacterial titer is important, as is the general ability of each animal to fight off a potential infection, and of course multiple exposures may be a factor too. Some animals (humans included) are very susceptible to a very minimal exposure, others may need multiple exposure to high viral titers and need to be in close proximity to the infected individual in order to become infected. So many variable can and do impact any viral transmission.

The other complicating factor is that CAE has the ability to infect, integrate its genome into the DNA of the white blood cell and become dormant for long periods of time (I use the 'cold sore' as an example - one day you're good the next day, blamo!) In the dormant period, the virus is not replicating, there are no viral particles for the immune system to recognize, so there will be no antibodies produced and the potential for a false negative from any test designed to detect antibodies against CAEV. The period of dormancy can be quite variable from weeks to potentially years. And as I've mentioned earlier, with so many animals being asymptomatic, or showing symptoms other than the encephalitis or arthritis, many breeder may not recognize an animal with CAE, thinking they have a chronic cough or prolonged sniffles, maybe showing some signs of mastitis, or simply listless behavior and weight loss. Without recognizing or appreciating these other symptoms, the risk of spreading infection can increase. One other consideration is that OPP in sheep is a close cousin to CAE, and each of these viruses is transmittable cross-species. so if you have goats and sheep on your farm, be aware that infections can pass between these animals. In more than one occasion I have had a producer tell me "no way can this goat be CAE positive - it has never been around an infected goat." Do you have sheep on the property? "Oh yes and that's where I keep the CAE-negative goats". Oops.

One final point I may or may not have made in the past is that in early stages of the infection, it is very possible to see test results ping-pong from high negatives to marginals or low positives and back to high negatives. Sometimes this can occur over the span of several months. this is probably due to the virus trying to get a good infective foothold with viral titers going up, followed by an increase in antibody titer. The viral titer may then fall as will the antibody titers, then the virus goes back into replication mode, viral loads go up and the antibody titer follows once again. At some point, the virus is going to win and the animal will completely seroconvert and begin to produce antibodies at very high levels and become a "true" positive.

Yes, there are many complications and considerations in the CAE world. Coupled with the fact things move slowly in this world as well. Regular testing of all the individuals in your herd is one of the best ways of combating the virus - find it early and take whatever appropriate measures you feel are necessary.

I hope this has answered some questions, and I'm sure has generated a bunch more. As always, please email me with your questions and concerns.

Thanks

Chuck

Sent from my Barcalounger


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## LLB101

ChuckAtBioTracking said:


> ... I would suspect overall in North America the infection rate could be as high as 75% since there are a lot of breeders who figure it's so hard to eradicate and most animals are asymptomatic...


Wow, I hope I'm misunderstanding this! Are you really saying that 75% of tests come back positive, that its still that pervasive? I just hear so much about everyone testing and being completely negative herd...


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## fmg

Hmm, where'd my question go...I'll ask it again I guess...needed better wording anyway.

If a goat is showing negative on a CAE test because she hasn't seroconverted yet, is she contagious?


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## Island Creek Farm

Does it matter which vials...just vaccuum or w/the clotting agent?


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## ChuckAtBioTracking

Guess I better weigh in here to try and correct any "missed understandings". I don't think I said that 75% of the goats we have tested come back a CAE positive. I'm in the process of compiling last years' data to see what that percentage is. There have been a few numbers tossed around (not just in this forum but in other articles as well) that anywhere from 75% to 90% of all the goats in North America are "probably CAE positive". So far I have seen nothing to indicate that is the case. I think it is true that there are many owners and breeders who feel CAE is so widespread and so many of the infected animals are asymptomatic that it is not really a "problem". They don't test, so we don't see what the actual numbers really are. Occasionally I will get a first-time tester with a very high percentage of positives. As they begin to implement proper herd management, the numbers on some of these farms goes way down over time. Others I never hear from again. Maybe this helps to clear things up a bit. Or maybe I'm not getting the point quite yet.

As far as the tubes are concerned, the standard red-tops are what I suggest. The blood clots fine, but if you have the clotting agent they are fine too.

Chuck


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## ChuckAtBioTracking

OK I take back one of my statements. In early September I did say "I would suspect overall in North America the infection rate could be as high as 75% since there are a lot of breeders who figure it's so hard to eradicate and most animals are asymptomatic that there really isn't that big a need for prevention." And in my previous post I think I said about the same thing - those who test regularly and act as proactively as possible with their herds will see a low percentage of positive animals on their farms. The only way to get any kind of valid number would be to get non-testers to start testing - if even only once!

Chuck


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## Caprine Beings

Hi Chuck!
My Vet (Dave Rustebakke) and I just drew blood on 11 goats. Three of which were on the pregger/CAE test form and the rest are on another form just for CAE. So our question is did we number them wrong? We started out with the three pregger/CAE testings at 1-3. And then we restarted numbering at #1 on the CAE form. 
What is the easiest way to do multiple forms? 

Tam


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## Red Mare

Also, what is the best day/way to send blood for CAE testing?


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## ChuckAtBioTracking

We find it least confusing to put all of the preg/CAE animals on the preg submission form, and check or write "CAE" in the Added Test boc on the preg form. For CAE-only, use the CAE form. Numbering the tubes doesn't seem to be a big issue since we send out two different reports. If you want to number the preg/CAE 1 2 3 4 and the CAE-only 1 2 3 4 that works. Or if you want to do preg/CAE 1 2 3 4 and the CAE-only as 5 6 7 8 we can handle that too.

As far as sending, during the cool weather, ambient air temp is fine. Summer months you may want to add a cold pack and do the o/night or 2-day FedEx. If you call or email us with the box dimensions and weight we can email you back a FedEx shipping form at 40% of what FedEx quotes you, and just add that to your monthly statement, or we can let you know the charges and you can add that to your enclosed check.

Using the regular mail can be tricky. Priority mail doesn't mean much except they charge you more for the same handling through the system. I think the only exception is if there is a Monday holiday, Priority gets priority over everything else and it can back up the system for several days. Also we are running into the holiday season so you may want to add 2 or 3 days onto your US Mail shipping schedule just in case.

We are on a 2-day per week preg schedule (Testing on Tuesday and Thursday with reports on Wednesdays and Fridays) and CAE is run on Friday, with the reports going out late Friday afternoon (so if you get your preg report on Wednesday or Friday morning and you had CAEs being run, have patience - we didn't forget!). This week CAE is on Thursday since we are closed on Friday, and preg reports will go out on Wednesday.

Good thing you didn't ask me a complicated question, I may have had to give a lengthy answer !

Chuck


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## ChuckAtBioTracking

CAE-positive goats (and OPP-positive sheep) who have not sero-converted can pass on the virus. Viral titers are probably low, so you may dodge a bullet on a single exposure (such as catching a kid starting to nurse off a low-titer but positive dam). Or you may not have dodged it. Tricky stuff this CAE virus. 

It is also possible that a dormant virus with it's genome integrated into the white blood cell DNA can pass the virus. Again, we're looking at low virus counts, but potential for a problem.

I think I mentioned this previously, but CAE-positive goats can pass the virus to sheep and OPP-positive sheep can pass the virus to goats. Also I have heard that one down-side to drinking milk from a CAE-positive goat is that you could test positive on an AIDS test. I'll do some more checking on that. And no, it doesn't mean you have or will get AIDS, it's just that the coat proteins for the two viruses have very similar protein sequences and, depending on the antibody used in screening, it may cross-react.

Chuck


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## DostThouHaveMilk

Chuck,
When you are determining the numbers of positives for records information (like the 75% tossed around), are you going back through and seeing when a known positive is retested? Or do all positives from one herd (even if they are the same animal twice) get counted individually.
I ask because I always retest positive animals. I actually just sent off 5 blood samples from 5 does who have tested positive on our last test (in the case of 4 and the other doe tested positive at her prior home). I automatically test positives a second time to be sure. So if they are counted each time they are tested positive rather than as one positive goat in the herd of 100, it skews the results for my herd and the overall records.
Am I making any sense?


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## ChuckAtBioTracking

As I go back through my data I am attempting to sort so rechecks are not counted. It won't be perfect but it should give a general idea of what we see from the more progressive breeders.

Chuck


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## DostThouHaveMilk

ChuckAtBioTracking said:


> As I go back through my data I am attempting to sort so rechecks are not counted. It won't be perfect but it should give a general idea of what we see from the more progressive breeders.
> 
> Chuck


 Wonderful! Thanks so much! Did you guys ever decide if you were going to offer a discount for larger herds? I asked before sending in my 85 to be tested this summer and you said you were considering it.


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## ChuckAtBioTracking

Unfortunately the powers that be in sales and accounting tell me we have the best deal around, and no plans in the near future for upping the price!


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## Caprine Beings

Anychance Bio-Tracking would be interested in adding CL to the testing roster? :lol
Tam


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## ChuckAtBioTracking

Working on it. The problem is most of the current CL tests work on one determining how the toxin affects red blood cell clumping (the IHA or hemagglutination assay) or measuring an antibody response to the toxin (the SHI or synergistic hemolysin inhibition assay). The sensitivity of these tests may not be high enough in some infected animals to give an unequivocal result. The best method is to culture fluid from the abscess in order to identify the organism, and we are not set up to do bacterial cultures (at this time). Plus I'm not too eager to culture and expose myself to a live pseudotuberculosis bug - hopefully for obvious reasons!

I would like to develop a highly specific ELISA test, but the assay would be based on using the coat proteins of the bug to detect antibodies in the serum, and again I'm not to hot on the idea of growing the bug to isolate coat protein to develop an assay. I do have some ideas on using some genetic engineering techniques to make a specific part of the coat protein to develop an ELISA, but research $$ are hard to come by these days. Unless some national caprine association has a few thousand laying around looking for a good cause 

Chuck


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## LLB101

I recall reading on the BioTracking site, but now can't find it, that preg test blood not only needs no refrigeration but there's also no rush on getting it there, that a few days in the mail is ok?


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## ChuckAtBioTracking

I was going to post this as a PM but maybe it may be valuable for several of you. If you want WADDL testing done on any of your serum samples, we can fill out the submittal form and shuttle it over to them for a $10 fee (one tube or 100 tubes). If it saves you on tubes, packaging and mailing it may be worth thinking about it. Just put a note on your preg or CAE form letting us know what you want. We don't know what their testing days are, so you may have to be a bit patient. And they will send the results to you, not us.

And yes, for preg testing, time in not critical. If you want CAE run too, it still is not a big problem when the temps are cool (70's or so) but in the summer months, if your package is delayed you may want to put a cool pack in and maybe think about 2nd day FedEx if you are a few states away. Please us the regular shipping cold packs - ice in a baggie leaks!

Again, we can offer a 40% discount on FedEx so email me for the details.


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## NubianSoaps.com

Chuck can you find out if they do the CL testing with SHI like UC Davis? For years WSU shuttled your blood to UC Davis, and now that they are running the testing is it exactly the same test? Thanks! Vicki


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## ChuckAtBioTracking

It is an SHI type test. I don't know who supplies the test to either lab - it may exactly the same. I do know they continue to forward sample to UC Davis for bacterial culturing and identification.

Chuck


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## LLB101

What's SHI ?

Seriously, someone else emailed me, they are too embarassed to ask, and apparently I'm always game... :LOL

Sure, I could probably Google it and figure it out, but the "chicken" folks are asking silly me to ask for public clarity for all. Apparently they have more pride than curiousity like I do, ehhheheeee. Good post to houseclean and just add the info :LOL


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## lovinlifeathome

synergistic hemolysis inhibition

Here ya go:

http://www.ag.ndsu.edu/pubs/ansci/sheep/as1353.htm

~Carla
JustRSize Acres
SW Oregon


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## NubianSoaps.com

LLB101 asked: When is EPT "pee on a test strip" coming for goats so preg tests are easy?

ChuckAtBioTracking answered: A urine-based pregnancy test is probably not likely. The human test is based on detecting the presence of HCG or human chorionic gonadotropin, produced initially by the developing fetus and later, by placental tissue. It is cleared from the serum through the kidneys and ends up in the urine. PSPB is cleared by the liver, so a urine test is not possible. It is present in milk, although at much lower levels (at least in cows (and yes I am working on it)) so for the current time we are stuck with a blood test. I have been working on an at-home 1 - 1.5 hr ELISA with the reagents in dropper bottles. The idea would be to put 5 - 10 drops of blood in the well, add a few drops from Bottle A, wait 45 minutes, wash it out with a squirt bottle of wash solution, add a few drops from Bottle B, wait, wash, add a few droops from Bottle C, wait, wash, then add a few drops from Bottle D and wait for a blue color to show up (pregnant).

The other idea is the Pregnancy Lolipop - a glass or plastic bead on a stick which you sequentially dip in the blood, wait, rinse it off, dip in solution A, rinse, etc etc to solution D and wait for the blue color to develop (or not, whereupon you are sad . . .)

So, despite rumors to the contrary, I'm not idly twiddling my thumbs, waiting for your samples to arrive or your phone calls to come in (smiley face)


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## hsmomof4

So, do the range of results in a pregnancy test tell you anything beyond "pregnant" or "open"? For example, I just got results back on two goats and they are both pregnant, but their numbers are different. So one is .6605 at about 60 days bred and the other one is .8281 at about 30 days bred. Do higher numbers potentially mean more babies, for example, or does it not mean anything at all?


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## ChuckAtBioTracking

The ranges are indicative of different physiologies of individual goat. I liken this to the differences in "normal" cholesterol reading between different people. I did do a study a few years back with a mixed breed herd of around 150 goats looking for a correlation between OD values and number of kids (they did ultrasound when they pulled the sample). Short version - no statistical difference between OD's from goats with 1, 2 or more kids. No real difference between breeds or size either, although in some boers their numbers were a bit lower - may be a dilution thing because of the greater blood volume of the larger breed but that's just a guess.

Normally we see OD's running from 0.4 to 0.8 which we consider normal. If I see a reading running in the Pregnant category but at say 0.25 or so, I suggest a confirmatory recheck IF the sample was pulled 30 - 40 days AFTER the buck was removed. If the days post breeding in a case like this is 60 or 80 days, this looks like embryo death and all of the PSPB has not yet cleared from the serum. A recheck is optional here because of the gestation time (if it is accurate as we define days post breeding i.e. NO potential contact with a buck).

I know a lot of breeders leave the bucks in all the time and start counting from a witnessed breeding event. This is fine, but the caveat is, if she comes back open she may be, or Mr Slick did the deed 15 days before you pulled the sample at 3 AM when you weren't looking and THAT'S the one that took!

Chuck


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## MF-Alpines

"We are on a 2-day per week preg schedule (Testing on Tuesday and Thursday with reports on Wednesdays and Fridays) and CAE is run on Friday, with the reports going out late Friday afternoon (so if you get your preg report on Wednesday or Friday morning and you had CAEs being run, have patience - we didn't forget!)."

Chuck: Are you still on a 2-day per week schedule? I was hoping for preg results today (assuming you received it on Monday or Tuesday). I was really hoping for results tonight. I waited all day yesterday until I realized that yesterday was Tuesday!


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## LLB101

I emailed them with the same question and didn't get a response either. On their website, it said they were closed yday, that's all that's listed for goats schedule affected by the holidays, but I too wanted confirmation.


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## [email protected]

CAE viability ?... It came up on a post in HT. 

Wondering if there's been any studies done on how long CAE lives in milk/blood? Have always heard that it doesn't live very long outside the body in blood...but obviously, it lives long enough to put into a vacutainer and ship to be tested.


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## buckrun

They are not testing for the presence of live virus.
They are testing for antibodies produced by the body of the animal.
Lee


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## KJFarm

We have been told that once dried (milk or blood) the virus is inactive.


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## ChuckAtBioTracking

Yes, we are still running on Tuesdays and Thursdays with results out on Wednesdays and Fridays from 1 November through 1 March. Be aware that the post office schedule is way messed up over the holidays so what normally takes a few days can take many "few days". We can provide you with 40% off FedEx o/night, 2-day or ground. Email me for details


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## ChuckAtBioTracking

The CAE virus is fragile and doesn't do well outside of the host. Once the blood/milk/mucosa/fecal pellet dries out, the virus is pretty much dead. I would still encourage people to treat any suspect areas with a 10% bleach solution.


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## ChuckAtBioTracking

I'm not aware of any studies directed at determining the viability of the virus in milk or blood. My guess would be that the CAE-positive blood samples I have in the cooler would probably contain infective virus particles for quite some time (months??). Contaminated milk may be the same, particularly if it is kept cool. The virus infects peripheral blood mononuclear cells (PBMCs) but think of them as "white blood cells" if you like. As long as the integrity of the cell is maintained, the virus is probably capable of being infective. Dessication will eventually destroy the cells, probably killing any live virus as well. Bleach effectively destroys the cells and the cellular DNA which contains the viral genome. This is part of the infective and reproductive cycle of the virus - it infects, integrates into the white blood cell DNA and uses the host cell machinery to replicate itself. As I have pointed out before, it is not uncommon for the virus to integrate and become dormant for months or years. This is sometimes why many people are surprised when a "negative" goat suddenly turns up positive after many rounds of testing and no know recent exposure to and CAE-positive animals.

I guess the bottom line is, assume any suspect milk/blood/etc is capable of being a source of the virus.


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## MF-Alpines

"True: humans cannot get CAE. However it may be possible to test positive for an HIV test if tested within a month or so of drinking milk from an infected animal. May be important if you are a blood donor."

Chuck: Can you elaborate on this statement? Can you also site studies?


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## ChuckAtBioTracking

I'm not aware of any published studies, but I'll search PubMed and see what if anything I can find. Based on the structural similarities between the HIV and CAE viruses and some of their biological similarities (methods of cell infection, viral genome structure and replication mechanisms) it seems possible that CAE antibodies could recognize HIV proteins.

If you are having your blood tested for whatever reason, and an HIV screen (looking for antibodies against HIV) is part of the testing procedure, CAE antibodies in your serum could react. It would all depend on how much milk you drank, what the CAE antibody titer is in the milk, when you drank it (the antibodies will eventually clear from your serum justas in unweaned kids raised on pasteurized CAE-infected milk), and the nature of the HIV screening procedure.

I would say the whole scenario is highly unlikely, but it is a possibility. Similar for example, to the possibility of having a positive urine test for drugs if you have recently eaten a poppyseed muffin. But let me be perfectly clear that drinking milk containing CAEV will NOT give you CAE and MOST CERTAINLY WILL NOT GIVE YOU AIDS!!


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## LLB101

Can you say a quick something about CAE numbers that are super low, well into "negative", but still why would one goat be different than the others that are all zero?

Ok, I found part of the answer in this thread
http://www.dairygoatinfo.com/index.php?topic=21670.msg0;topicseen#new

Where it say lots of things that generate an immune reaction can get a number over zero, but of no concern, usually its recent immunizations?

In her case, she's had no immunizations, is a FF, was 46 days post breeding when the blood was drawn, and seems super healthy.


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## ChuckAtBioTracking

the simplest explanation is that everyone is different as far as their biology/physiology is concerned, so everyone will respond differently to any given test (goat, cows, horses, humans, you name it). If you look on the pregnancy reports, no two goats will have exactly the same OD reading (well if you look hard enough you may find one LOL), and even for one goat, the OD values will be different for each sample drawn. This is because PSPB is constantly being produced by the placenta and cleared by the liver, but the overall concentration remains fairly constant.

The CAE test is a different assay - still an ELISA but a much different format. The percent inhibition levels are calculated from the control samples I run on each assay. The negative control, for reasons on how these types of assays work, is not set at the lowest possible value and is in fact somewhat positive in its response (relative to a "true" 0.00000). A lot of CAE-free goats have raw data scores that are less than the negative control (a good thing) and actually give a negative percent inhibition. I have the reporting software set up so a -9.5% is scored as 0.0, so all goats with negative values are set at 0.0. I thought this would be less confusing than reporting negative numbers.

One thing I want to point out is that as long as your goat is testing negative, he or she is negative (as of the time that the sample was taken). So if you have 4 goats that are 0.0% and one that is 14%, it's still negative, just a higher basal level than the others. And those basal levels can change over the life of the animal just like out blood sugar or cholesterol or whatever is being tested on us, will change over time. The critical thing is that the test level falls in the "good" range. So don't be culling your goats testing higher than 0!!

As I have said before, I don't get too concerned about things unless a goat scores in the mid to upper 20% range. There may be something going on there (early stage seroconversion? interfering substances in the serum? just his or her "normal" range for the assay?) so I try to call it to the client's attention and suggest a recheck in 4 - 6 weeks to see if things are changing at all.

One thing I should point out (it may be in another posting somewhere) is that in the early stages of infection/seroconversion it is possible for a goat to have widely varying responses to subsequent tests. I have seen several animals go from 28% to 37% to 22 % to 42% and ping-pong like this for quite some time. I think this is due to an increase in CAE antibodies as the viral titer increases, a drop in the viral titer in response to the antibodies sort of successfully fighting the infection (they will eventually lose, but they try hard at first). This causes the viral titer to drop, so the antibody titer drops, causing the virus to mount another battle. Virus titer goes up again, antibody titer goes up again. Back and forth until eventually the virus wins and the antibody titers zoom up to the (in the seventy to 80 percent) or so percent range and stay there for life.

Sorry, but I do tend to go on about things! Does this help any?


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## LLB101

I like you going on about it Chuck! 

In this case, it wasn't the info I was looking for exactly. This was more about borderline cases, retesting etc which is great info for the files.

My question was more about the clearly negative result, but one that still registers a number. You do mention a "basal" level here, any ideas what influences those differences? Other threads have mentioned recent immunizations, in her case I said she hadn't had any. I think her number was 14 if I remember right, I'm not worried about her starting any kind of seroconverting, I really don't think she's had any opportunity for exposure, its well below the "negative" threshold etc. I'm just curious. :biggrin


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## ChuckAtBioTracking

Those that are CAE-negative but with a value (14%) are just that - negative with a number. But don't get the idea that the same animal will have the same value from test tot test. It would be nice, but neither science nor the animal is quite that cooperative. There is always "drift" in the test as the lot numbers for the reagents change, and just minor variations in manufacturing of the assay plates and reagents. So I don't get too excited about numbers varying from 7% to 12% to 4% for the same animal. It's the 7% to 25% that would raise a flag, and I would suggest a retest on a fresh sample in about a month or so. If I think something went funny with the assay, I'll retest the original sample just for confirmation


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## fmg

Chuck, what is the point of putting "exempt animal specimen" on the outside of the box when shipping blood to the lab? It seems some of the people here are having trouble with it shipping promptly with that label attached (or perhaps it is an unrelated problem).


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## ChuckAtBioTracking

It's not the exempt sticker really, it's the HAZMAT sticker some postal employees insist on putting on the package. Yeah this is a prime example of how the govt bureaucracy doesn't work.

Postal regulations forbid sending of hazardous material (like blood) without an identifying HAZMAT sticker on it. For animal (and some human) diagnostics, blood is considered exempt from that regulation. Funny thing is, a lot of postal clerks don't even know about the exemption and want to put HAZMAT on it If you have problems, you may want to get the actual explanation from USPS:

http://pe.usps.com/text/pub52/pub52apxc_030.htm

take it in and show them (but even then some will not pay any attention to you. Sigh . . .)

It becomes more of a problem with Priority Mail (don't get me started on that oxymoron) since USPS contract with FedEx or UPS for air transportation, and they have strict policies about HAZMAT stuff which needs to go ground. It's actually a problem with any mail no doubt since almost all of it goes by air at some point in time.

I guess it all boils down to how truthful do you want to be when they ask "Does it contain anything hazardous?" (no and that is truthful). "Any liquids?" (well . . . it IS clotted, but there is some liquid as serum . . .) So a slippery slope. Got your ice ax handy?


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## ChuckAtBioTracking

The BioPRYN test for sheep and goats is available at other testing laboratories! Please check with them for pricing and scheduling.

http://www.biotracking.com/pdfs/Affiliates that offer Sheep and Goat 12-1-11.pdf


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## LLB101

Does the pregnancy hormone that you test for stay at high level thoroughut the pregnancy? The one of mine that is due in less than a month had the lowest levels of any I've tested. I wasn't sure she was pregnant, wasn't really looking very pregnant, but maybe. Belly did have that tight feeling that the pregnant ones do, and she hadn't come back into heat, so I thought she was, but tested when I was sending in others anyway.

Results came back "pregnant" but with the lowest levels of any of them (0.5 vs 0.8 &9 for the others). She's the furthest along, due in 1.5 months at the time of the test, if she was pregnant, so 110-ish days post breeding. The others were in the 30-50 day post breeding range and I was testing to make sure since we're running out of normal breeding season.

In the past couple weeks since the test, she has suddenly started looking pregnant, I'm just curious about the levels at different stages.


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## ChuckAtBioTracking

For sheep and goats this does seem to be the case. For cows, after about 90 days, PSPB protein levels start to go up (and this is reflected in the OD values up to a point) and continue to increase right up to calving. Sheep and goats seem to come up right at around 28 days and pretty much stay constant for the duration. I'm pretty sure that in actuality, their PSPB levels are similar to cows, we just can't see it with the current antibodies.

I don't get too concerned as long as the OD readings are around 0.5 and up. If they run significantly lower than that, but are still in the pregnant range (ODs of 0.2 - 0.3), I try to alert the breeder and ask a few questions. There are two or three reasons for a Pregnant score with a low OD reading: 1) this may just be the case for this particular goat. She doesn't produce as much PSPB as most goats; 2) there may have been embryonic death and we are seeing declining PSPB levels as it clears, or; 3) the actual days post breeding is less than 28 days and we are seeing PSPB levels going up in a pregnant doe. Usually talking with the breeder can help decide what is going on, and in these low OD cases if the breeder want to retest in a week or so I generally do it at no charge. But at 0.5 I think you're OK 

There can be a lot of factors that come in to play as far as the PSPB levels for individual animals. Every ruminant will produce different amounts of PSPB according to what their reproductive physiology and genetics dictates. And larger animals can have lower amounts of PSPB (in terms of ng/ml of serum) since they are physically bigger with a greater blood volume which will have an effect of diluting the PSPB present. And there is probably some effect from multiple embryos, but it's not a 1:1 ratio, so there probably won't be twice as much PSPB for twins as for a single fetus. Sometimes we see this in cows, but a dairy goat study we did a couple of years ago, there was no significant change in PSPB levels for multiple embryos.


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## LLB101

Interesting, thanks!

She's significantly smaller doe than both her half sister with the 0.8 and her littermate sister with the 0.9 result. So that's opposite of what you're saying about size, right? We'll see what she has in 4 weeks, larger sisters aren't due until late March & mid-Apr.

What do we put on our order form if we want to retest one? I had that other one that was 0.114 which is close enough to the 0.127 since she was actually only 23 days? I put 26 on the form, typo in my spreadsheet.

Was at classes all day by WSU vets/PhD... very frustrating news about the state of QFever data and testing in light of the hysteria this year. Do you have one in the works that's any better? There's a good human test I heard.


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## fmg

Chuck-just had an idea. Since it is difficult for you to track a specific doe and see how long it took for her to seroconvert, I was wondering if you might put a box to check on the forms for whether this goat was previously tested with Biotracking? Do you all have any kind of database entry with previous results, or is it all on paper?


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## ChuckAtBioTracking

We have all of the reports on the server, but with the way or data base is set up now, we can track only by LOGIN number or owner/farm name. Yes, if you indicated on the form that you wanted us to look at a particular goats history, we can do that, but it seems easier for you to make a spreadsheet from the reports of each of your goats?? We can send the reports as both the HTML version and as an EXCEL spreadsheet, so you can cut-and-paste into your own spreadsheet.

Or am I missing something in your question?


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## fmg

Well, I am curious what the longest you know of that an animal took to seroconvert to positive...but I didn't think you have a way to figure this out, because you may not know that it was the same animal, but maybe I was wrong?


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## ChuckAtBioTracking

I don't think I have a way of determining this since I would need to know when the exposure date was. Maybe someone who has tested negative for years and suddenly had a confirmed positive pop up could think back on when a possible exposure may have occurred. I can ask the CAE gurus if they have any hard and fast data, or even an off-the-fly guess and see what they say.


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## ChuckAtBioTracking

WADDL TESTING

OK so a bit of a change in plans. In talking with the lab techs here, this is the procedure for requesting additional testing at WADDL:

1 - You must fill out the submission form and send it to us along with the samples. The general acession form can be found at

http://www.vetmed.wsu.edu/depts_waddl/forms/AcessionGeneralDiagnosticsWADDL.pdf

and with an update Adobie you can fill it out on the screen and print it out. It's a goofy form so what you need to do is ignore the Vet Info section on top and fill out the Owner Information section. Leave the WADDL Owner Client Number blank unless you have one. Check the Serology box (for CL and Johne's). If you have one animal to test, write the Animal ID in Specimens Submitted box, and I would indicate there and in the Additional History section what tests you want performed (there seems to be no other place to indicate what you want . . .)

If you have additional animals use the Identification Sheet Multiple Animals

http://www.vetmed.wsu.edu/depts_waddl/forms/IdentificationSheetMutipleAnimalsWADDL-014.pdf

and include that with the Acession Form.

Fee schedules can be found at

http://www.vetmed.wsu.edu/depts_waddl/immunoDX.aspx

and click on the Ovine/Caprine link just under the General Information section at the top.

We will then take the samples and forms over to WSU for the $10 one-size-fits-all fee. Apologies for the miscommunication. With the increasing number of requests for additional testing, the time it takes to fill out the forms for everyone becomes cost prohibitive from our end. If you have questions, please call or email me.


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## ChuckAtBioTracking

MayLOC said:


> Chuck
> Was looking for more info on the: BioPRYNQK® listed under cattle on the biotracking site.
> I read that it is a good test for same day results but couldn't find any more info on how you do it.
> thanks
> kassi
> 
> for one, I couldn't figure out if it is some sort of on site testing kit and you don't have to send blood to the lab at all and that is why it says you can get immediate results.


WE have developed a new set of reagents and antibodies that allows us to run the test with three 1-hour incubations (sample, secondary antibody, and detector). The OD values are going to look different from what you are used to with the 'normal' BioPRYN, but the report will still give you the cutoff and repeat thresholds. It's more expensive than the standard test ($3.50 v $2.50 - we just increased the standard test by a dime), but if you overnight your samples and want the results that afternoon, it's the way to go.

Does that help?


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## MayLOC

thanks Chuck. Yes that helps. So you still collect blood and send it to the lab same process, just a speedier test at a little higher price. We can't overnight things to you with USPS (I tried yesterday and they can only guarantee 2 day delivery from here) but I bet maybe Fedex could. We often preg test large groups of heifers and cows, but for the most part I guess it would probably make more sense to stick with vet preg checking so we only have to get them in once and sort them once and most of our cattle do not have tags so it would mean temp back tags also til we got the results. But for smaller groups or handfuls or an older cow here or there it could be a valuable tool for us to use. Always looking to streamline things. thanks.


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## ChuckAtBioTracking

Don't forget our 40% off the "list" price for FedEx. Call and ask for specifics, although there is a thread in here somewhere . . .


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## SALTCREEK_Nubians_Linda

This has probably been asked, but I need to know soon. I have a friend with a registered herd of Boers. She wants to buy a couple of my Nubians. She wants to know how much it will cost her per animal to test her herd of Boers for CL and CAE. We are hoping to get her a clean herd of both breeds. So far she has had no abcesses and her animals came from a breeder, not a sale barn. She has no swollen knees either. Can you give us this info?

Also, I am switching to Bio-Tracking I think. I want to CAE my Nubians and G6S tect my buck. Do I need packaging from you? A requisition form? Any special packaging I need to do? Please Inform. Do you test colostrum, and how do I prepare that? Thanks. Linda


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## ChuckAtBioTracking

Linda - CAE is $4.00 per sample, and is a serum-based test (red-top blood tube). CL testing is done through Washington State University WADDL. Check reply 66 above for the info on getting submission forms and requesting us to forward them to WADDL. You will need to send in about 2 - 3 cc of blood. Packaging and shipping can be found at

http://www.biotracking.com/?q=FAQS "How should blood samples be prepared and shipped"

Thanks


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## fmg

How common is it for a goat that is just above the "pregnant" range to retest as pregnant still? I have one that tested at 0.185 on 79 dpb. Is there anything (hormones, proteins?) that can trip the test and show a pregnant when a goat is not or has never been pregnant?


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## ChuckAtBioTracking

I would say that 95% of our pregnant samples result in a test response of between 0.4 to 1.0. Most are actually in the higher ranges. Those that come in at the 0.2 - 0.3 OD response are of concern and may reflect the slow rate of clearance of PSPB from the serum following embryonic death. In a few cases (one today from Florida) a goat tested in a high pregnant but recheck 7 - 10 days ago, then tested again this week in the same range. Based on her ultrasound (abnormal) and her past history of losing kids, this looks like an embryo in trouble, just hanging on, but still simulating small amounts of PSPB production. We have seen this in cows off and on over the years, and I don't think there has been a case of carrying a fetus to term. Sometimes these "troubled embryos" can maintain a recheck or low pregnant response for several months before dying, at which point the blood test a week or 10 days later will finally show as open.

At 79 DPB with that low of a pregnant response, I'd say she is either holding on to a trouble fetus, or aborted or resorbed in the last few weeks. The only other thing we have seen (and rarely) has been a batch of virgin heifer serum (actually from a steer) or a bull elk sample that tested with a marginal PSPB response. In both cases these animals were diagnosed with a reproductive tumor. It may be that these tumors were producing something that looked closely like PSPB, at least enough to cause some cross-reaction with our antibodies.


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## Halo-M Nubians

Chuck! Hi there. I have a question I'd love to get cleared up. 
Is a doe who is testing negative, but has a dormant virus, just hasn't converted yet producing contagious milk? This is concerning for those of us who choose to feed raw but test frequently. Obviously if she is a dormant positive we have no way of knowing at that point. I had always assumed that a neg test meant no contagious virus AT THAT TIME, at least. But below is a comment of yours that has concerned me that I am wrong in this assumption. Also is the info from your website which seems to contradict. Thank you so much for you willingness to try and educate us all on CAE!! any knowledge you can share is much appreciated.

From the website: (bold add by me)

Q. How often should a producer test for CAE?
A. It is important that producers understand that the CAE test is a "snapshot in time" and that the CAE virus can lay dormant for months or years *before it begins to reproduce and become infectious*. Therefore routine testing is important if you want to maintain your herd as CAE negative. It is recommended that all animals be tested every 6 months.

From a previous post in this thread:
*CAE-positive goats (and OPP-positive sheep) who have not sero-converted can pass on the virus*. Viral titers are probably low, so you may dodge a bullet on a single exposure (such as catching a kid starting to nurse off a low-titer but positive dam). Or you may not have dodged it. Tricky stuff this CAE virus.

It is also possible that a dormant virus with it's genome integrated into the white blood cell DNA can pass the virus. Again, we're looking at low virus counts, but potential for a problem.


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## ChuckAtBioTracking

Well, this becomes a possibility / probability kind of thing, with a lot of "what-if's" entering into the equation. Goats and sheep are probably going to be more highly infective if the virus is actively growing and producing a high titer of infectious virus particles. I'm not sure if milk containing infected WBC would result in disease transmission or not. I doubt WBC can pass through the intestinal tract so they probably just get excreted. The major problem are those animals that are actively infected but are not yet producing antibodies, or sufficient antibodies, to be scored positive in an ELISA or AGID test.


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## Halo-M Nubians

So if I understand what your saying..IN your opinion =) an negative testing animal is probably unlikely to be contagious through white blood cells in milk. But a borderline (converting) animal that may not be testing positive yet could be.


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## ChuckAtBioTracking

In my never-to-be-humble-but-still-CYA opinion, yes


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## ChuckAtBioTracking

I'm not aware of any published research on this, but let me do a search and see what I can dig up. But, since it is a lentivirus (related to HIV), it can enter a stage of dormancy after what might be considered an acute infectious stage. I'm thinking it may be possible for breeders to miss this stage given the following scenario: the white blood cells are infected; viral load increases; and probably followed by a rise in antibody titer. If the antibodies mount a successful response, the viral load goes down and the virus may enter into a latent stage. If this happens, the initial antibody titer decreases with time, and if they test after all of this has been going on, based on an ELISA or AGID test, you have a "negative" goat. HIV can be dormant for 6, 8, 10 years before symptoms of AIDS are evident.

My suggestion would be, if you have had CAE on your farm, culled positives, and have been testing negative since then, it might not be a bad idea to do a whole-herd screen by PCR to try and eliminate any 'sleeper' in the herd. If you have 20 goats at $25 per assay that's $500, but how much is your prize milker you may have to cull?

I'll post some additional info after I do some digging.


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## LLB101

Back to the pregnancy testing... The FF doe that had such a low number at ~4months pregnant, she kidded with a single, very small, not thriving. I think you said there's not a direct relationship with numbers of kids, but a low number can indicate a pregnancy in trouble, so this is an interesting result. The two that had high numbers 0.9-ish are huge already, so it will be interesting to see what they have in comparison. Someone was speculating that it might be related to the "total weight" of kids being made and supported, not the literal "number" of kids. I've tracked that here and bugged others for data, and kids seem to run in total weight, about 13-18% of the does pre-breeding weight. Over about 20% and it seems like there starts being trouble in paradise. Far from a scientific sample yet, but interesting...


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## ChuckAtBioTracking

I'm getting more and more data on these "Pregnant-with-low-numbers" and it is beginning to look like this does show embryonic problems. I tracked a couple of does last September/October who came in as Pregnant at around 0.2 (others were in the 0.6 - 0.8 OD range) and a few weeks later restested one as open and the other as still pregnant but seemingly lower PSPB levels, so I'm guessing she was still clearing from an EED, or still with a struggling embryo. 

I talked yesderday with another breeder, same situation at about 60 days out (AI no bucks around) for 2 of his does, and all of the others bred the same day have kidded, these two did not. So I'm still going to offer the free recheck of these types of situations untill I get the "new and improved" assay in it's final stages. I'll let you know now that the reports will look pretty goofy: my choices are to give the raw OD numbers which are going to be in the 2.0 to 3.0 range for pregnants; and around the 0.15 range for opens. The other option is to normalize the results so they look similar to what you get now (kinda sorta) but you may see a lot of pregnants with EXACTLY the same number and the opens with EXACTLY the same number. This is what happens at the uper and lower ranges of sensitivity of the assay if you throw a bunch of normalization math at the results. I'll keep you posted.


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## fmg

The most puzzling one I have heard of (and I think she called you...she said she did anyway), the doe was something like 40-50 days bred and came back open; her pregnancy was confirmed when she kidded 3.5 months later! Do you know of the one I am speaking of? She kidded toward the beginning of February, and had been preg checked mid-October. I've not heard of anybody else ever had this problem. The twins are perfectly healthy Nigerian dwarf goats.


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## ChuckAtBioTracking

fmg said:


> the doe was something like 40-50 days bred and came back open; her pregnancy was confirmed when she kidded 3.5 months later!


Hmmm . . . I don't remember talking with her, but I'd be surprised if she didn't call. Our specificity (truly not-pregnant goats identified as not-pregnant) is greater than 99.9% so based on that we're going to miss 1 out of every 1000 and someone has to draw the short straw. A good thing she was suspicious about it and waited her out! BTW when you suspect something may not be right in a report, make sure you let me know so we can arrange a re-test of the original sample and if needed, a retest of a second sample for confirmation. I run the Las Vegas odds on these too - if I'm wrong I pay, if I'm right you pay!


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## ChuckAtBioTracking

Another note on low pregnancy ODs. I took a call this morning from a vet whose client tested back in December, 60 DPB with a test OD of 0.62. They sent in another sample a month later with an OD of 0.487, then again at 130 days with an OD of 0.242. It doesn't look as if I called any particular attention to the 0.487 reading since it wasn't that low (but in hindsight at 90 days maybe I should have scratched my head a bit . . .) If I knew this was a second test on this doe, I would have looked at the first report which would certainly have raised a flag. When we get the new software debugged, we'll have the ability to enter in an animal name or ID along with a farm name and get the results of any testing we have done over the past 10 years. So until then, if you think of it, add a note saying "I tested her a month ago" and I'll be sure to look up the history.


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## LLB101

ChuckAtBioTracking said:


> Another note on low pregnancy ODs. I took a call this morning from a vet whose client tested back in December, 60 DPB with a test OD of 0.62... a month later an OD of 0.487, then again at 130 days with an OD of 0.242. ...


I'd love to hear if/when she kids what she has and what evidence there might be of another EED besides the surviving kid(s) if any. Are you mentioning to them to look for possible evidence, that there is this theory?


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## ChuckAtBioTracking

LLB101 said:


> ChuckAtBioTracking said:
> 
> 
> 
> Another note on low pregnancy ODs. I took a call this morning from a vet whose client tested back in December, 60 DPB with a test OD of 0.62... a month later an OD of 0.487, then again at 130 days with an OD of 0.242. ...
> 
> 
> 
> I'd love to hear if/when she kids what she has and what evidence there might be of another EED besides the surviving kid(s) if any. Are you mentioning to them to look for possible evidence, that there is this theory?
Click to expand...

Forgot to add to the story that the doe delivered one stillborn kid.


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## Rockinddtoggs

I have been wondering if the condition of a doe can have any effect on her titer levels for pregnancy. I have a doe we brought into the herd in December, she was almost emaciated but supposed to be bred. I sent you the test and it came back low at .29 at 60 ish days along. I never rested her and she gained some weight back and is looking better. She even kidded triplets lastnight. All are healthy and look good but could her bad condition have made the kids weak in the 1st part of her pregnancy and caused to low level?


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## ChuckAtBioTracking

Rockinddtoggs said:


> I have been wondering if the condition of a doe can have any effect on her titer levels for pregnancy.


I don't know for sure, but I wouldn't be too surprised. We really don't know too much about what theses PSPB proteins really do as far as pregnancy is concerned. They are members of a group of proteins called aspartic proteinases, and function to break down other cellular proteins. There is probably a very complex biochemical "dance" involving the PSPB family of proteins, the type of proteins they degrade and the times in pregnancy at which they degrade them, and other proteins that regulate the presence of the PSPB proteins. We do know they are produced in highly specialized cells in the placenta, and production is probably stimulated by specific biochemical signals from the developing embryo. It stands to reason that if the embryo is not healthy, production of the signalling molecule may be reduced, in turn affecting the production of PSPB. As mom gets healthy, the embryo becomes more healthy, and eventually PSPB production could increase to 'normal' levels.

Good thinking!


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## LaManchaLady

Chuck, 

I was asked this question and I honestly dont know. A friend of mine told me she saw on Airtite where they had the plastic testing tubes and was considering getting them; but was wondering if there would be any difference in test results from a plastic test tube and the glass test tube. I have always used the glass tubes myself so I was of no help. Thank you so much in advance!


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## ChuckAtBioTracking

Plastic is fine. The caps are a bit different, and we have seen a few that are cracked (probably from shipping) but there is no effect on the test. This is true for purple top, green tops, serum separators etc.


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## AJ

ChuckAtBioTracking said:


> We will then take the samples and forms over to WSU for the $10 one-size-fits-all fee.


What do you mean by one-size-fits-all? I have 5 goats which I'd like not only CAE tested, but CL tested as well... Do you mean $10 for each goat specimen covers only the CL test, or both CL & CAE?


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## fmg

$10 is the delivery charge for Biotracking to take samples to WSU, as many samples as you want them to. WSU will have charges per sample to test them for CL.


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## kuecktr

Has a goat tested positive for CAE ever tested negative again? I had a doe test negative twice for CAE as a yearling and then test positive a year later  Thank you, Taylor


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## Halo-M Nubians

Taylor, if you read back through this thread I believe there is some discussion about how the virus can lay dormant for up to years..I think chuck refers to that as "sero-negative" but the virus is there just dormant. This can explain how exposure as a kid doesn't pop up right away and you can get those neg tests then some stress factor causes the virus to "wake up" so to speak. This is why continual testing is important. I would always double check a positive result to be sure it wasn't a mistake. But once positive there is no going back.
Sorry! I know I'm not chuck! -lol!


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## LLB101

... can you tell the diference in a pregnancy result if the doe is 29 days or if she's 50days pregnant? That's not very clear question... LOL.

We thought she was bred, then came home to find her in the bucklings pen on the day she would have been in heat again. They were 5 & 6 months old so old enough to breed too and confuse the issue. But maybe not big enough unless she was very motivated to help LOL.

I needed to get blood in right way at 30 days past the first date, but well, it didn't happen.

Suddenly its now 29 days past the "pen party" date, yikes! 

So I did get blood drawn on day 29... will you be able to take an educated guess if she's pregant at 29 days or at 50 days, is that a more clear question now? .


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## ChuckAtBioTracking

To answer your question, the simple answer is no. Goats and sheep seem to produce PSPB at the same levels from day 28 out to kidding, and there is some variation between the levels in different breeds and sizes of does. Even with cows it's difficult to make any kind of DBP determination in that time window of 28 to 60 days since the PSPB levels are pretty much level. Around day 70 or so we can start to see an increase, and it continues to do so right up to calving.

Here's what I think is happening, for those of you who are curious about the biology / biochemistry side of things. For cows, there have been around 20 different "flavors" of PSPB that have been characterized. For sheep and goats I think 6 - 8 have been identified. Even though no one knows what these proteins do, there is some evidence that some are produced early in pregnancy, some later, and some for the duration. These belong to a family called aspartate proteinases which means they are proteins that eat up other proteins. It's possible their function may be to eliminate some other proteins at a particular time once their role in fertilization, implantation, stabilization, maintenance etc etc are over. Maybe a kind of pregnancy garbage disposal!

For cows, there seems to be a "changing of the guard" at around day 60 or so since there is a bit of a dip in the PSPB levels, This may be a result of the early PSPBs being degraded and cleared, and the late-term PSPBs starting to increase. I suspect this is true since we have been able to develop some antibodies that recognize the early stage PSPBs better than the late-stage, and some that do the opposite. I think the same is true for sheep and goat as well - different "flavors" at different times, but the antibody tools I have for sheep and goat seem to be directed at those PSPB that are pretty much produced at a constant level throughout pregnancy. And yes, I've been looking for other antibodies that may be able to differentiate all these guys, but nothing of significance yet.

I recently finished up a study with Dr. Reid Redden at NDSU looking at PSPB levels in 4 different breeds of sheep starting at day 24 and going through day 60, with the idea of trying to identify multiple embryos. Although we could see some slight increase in levels from day 24 to 60, and some correlation between PSPB levels and number of lambs, I'm not sure anything was statistically valid. We'll continue to work on identifying other antibodies that may give us more definitive answers, particularly when it comes to the question of multiple embryos. Knowing this would certainly be helpful in monitoring differing nutritional needs between singlets and multiples, particularly with feed costs being what they are. I'll keep you posted as (if) we progress!


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## ChuckAtBioTracking

Merry Christmas and Happy Holidays to all! I'm planning on releasing the new sheep and goat assay after the first of the year. The most significant thing you as producers will note will be (hopefully) a decrease in the number of rechecks on the reports. This should also reduce the number of "false" positives - those animals that are reported as pregnant but with significantly lower OD readings than you see for a normal healthy mom-to-be. For those of you who have fallen into this category, it would be helpful if you could let me know the log number of the report, and what the final outcome was for that "low OD pregnancy". Unfortunately we are still unable to give any accurate number for days in gestation or number of kids/lambs, but if you really insist on a number I suppose I can give you one . . . :biggrin

For the affiliate labs running the assay, the kit is now a series of 1-component buffers and reagents - no more mixing of things, and you will be running a different report-generating program. Let me know if you need an update on that software.

Thanks once again for all your support over the years, and keep looking for new products to come along (Johnes testing is still in the works stage but I remain ever hopeful. CL testing, well, not so much . . . )

Chuck


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## Island Creek Farm

Just curious, I sent in blood for pregnancy testing and CAE and only got the pregnancy results in on Friday (it's Sunday)...do the CAE test results get emailed separately, and at how much of a delay? Thanks!


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## ChuckAtBioTracking

Hi Amanda - For your (and everyone's info. Things have changed a bit since I have stopped overseeing the sheep and goat pregnancy and CAE assays) we are currently running the pregnancy tests on Tuesday and Thursday with results sent out on Wednesday and Friday. If your samples arrive in the lab on Tuesday, they will be run on the Thursday test. If they arrive on Thursday they will be run on the following Tuesday.

CAE is run on Tuesday with the results sent out that day. If your samples arrive on Tuesday, they will be held and run the following Tuesday (sorry the timing is completely out of my hands now). Amanda, you should get your CAE results tomorrow, Tuesday 15 January 2013 around 5:00 PM PST. If you don't receive it, email me or give me a call.

Thanks

Chuck
208.596.8776


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## Annie

Chuck - just heard from my local vet - she'll be here early Friday morning to do blood draws for me. I want to send them to Biotrack for CAE testing, can you get me started?


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## ChuckAtBioTracking

You will need to have 2 - 3 cc's of blood drawn in a standard red-top tube (4 cc is better if you use the serum separator tubes). Make sure to use a fresh needle on each animal - I know that seems like a no-brainer but I've heard stories . . . ). Bundle the tubes together (groups of 5 work well) with a rubber band, place them in a ziplock bag, and box them up with packing peanuts, newspapers, NOT old underwear (yes it has . . .) and ship them off. USPS will tell you "It should be there in a couple of days". They tell everyone that about everything - it is NOT a guaranteed delivery. You can go UPS or FedEx - call us and talk to Lorri and she can set you up with FedEx at a 40% discount. You will need to know the size and weight of the package. Do NOT put, nor allow the post office if you use them, to put a Biohazard sticker on the box. It's best to write EXEMPT - ANIMAL SPECIMEN on the outside. Here's hoping for negative antibody results!

Chuck


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## [email protected]

Annie--I am in NE Indiana and have always shipped USPS WITH delivery confirmation. It has ALWAYS been there in two days.  I went and helped a friend draw blood on Wed before Christmas. She mailed it that day with delivery confirmation cuz it had to be to Biotracking on Fri to get tested Monday (Christmas was Tuesday). Even with holiday mail/shipping, she had her results on Monday, so it made it there in two days.


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## fmg

ChuckAtBioTracking said:


> NOT old underwear (yes it has . . .)


If we send it with really sexy frilly underwear, do we get a discount? Chuck, what is your panty size? :rofl :laughcry


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## ChuckAtBioTracking

Sent you a PM! :biggrin :lol


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## MF-Alpines

Nancy, you are too funny! Chuck, too!

I have stopped putting the EXEMPT ANIMAL SPECIMEN on the outside of the box because last year I had two shipments delayed for over a month with USPS. I do put the Biohazard label in with my specimen for BioTracking, but that is it. When they (USPS) ask me if there is any liquid, hazard, blah, blah, blah, I just say nope, nope, nope, nope, nope. We have a small post office. I think they know something is not going according to USPS regs, but they don't hassle me. Unless I have the EXEMPT ANIMAL SPECIMEN on the box!

I've thought about what I would say if they asked what was in the box. I'm going to say "hair samples". Then tell them a story that you can get disease and pregnancy confirmation through hair. They'll never know.

That's my story and I'm stickin' to it!


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## ChuckAtBioTracking

Can't say I'm too surprised since it does seem to happen fairly often. A bit odd I think since it IS a postal regulation:

"Reference DMM 601.10.17.9
Exempt human or animal specimens as defined in 346.12d are not subject to regulation as hazardous materials, but when presented for mailing, they must be properly packaged. See 346.326"

Here is the link if you want to read the full explanation. Seems like it doesn't pertain to CAE, Johnes, CL, BVD etc so, wearing my CYA hat  I'd say you can't use that label for those testings.

http://pe.usps.com/text/pub52/pub52apxc_030.htm

Chuck


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## fmg

Chuck, when we spoke on the phone you said that a pregnancy test with an OD of 0.5-0.8+ was generally great, but below that you would start to worry a bit (or something similar, that was my impression). I just got a test back on a doe at day 31 post-breeding, and her OD is 0.3246...should I be concerned? This doe is AI'd, so I would like to be sure her breeding took.


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## ChuckAtBioTracking

Most of the does on that day were running in the high 0.8 - 0.9 range. There was one other at a 0.28 and showing pregnant (the threshold for this assay was 0.21, anything below that was scored as a recheck). I think I'd send in another sample in a week - mark it to my attention so it will be run under Research and Development and not under Lab Services. I'll run both samples so we can get a direct comparison between them. The values may not change, which may mean your doe is just running lower PSPB levels. Is this a large Boer by chance?

Chuck


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## fmg

Nope a smallish alpine. I will draw and send on Monday. I'll mark the Log # from the previous test on the form. Do I pay for this test, or will it be a no-charge re-check? Just so I can be sure to get the money pre-paid, if needed...I'm bad about paying my paper bills...


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## Goat Town

What is the earliest age a kid can be tested for CAE if it has been raised on heat treated colostrum and pasteurized goat milk?


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## NubianSoaps.com

Nicole I test all my purchased kids, even 3 week olds. If they have received CAE positive colostrum or positive colostrum that has been heat treated their titer will rise. Since I only purchase from those who are long CAE negative and whole herd test, or advertise as such, there would be no reason for a borderline or positive goatling, I would demand my money back. Vicki


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## Goat Town

Thanks Vicki,

Sionce I now know the CAE status of my herd I'm considering not selling kids without testing them first.


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## NubianSoaps.com

Even though I am long negative, I still sell on contract and give new folks the chance to test via biotracking their goats or kids. For myself that was the best way to do this. To get another kid though a vet does have to pull the blood, read tattoos and send the blood themselves, then their vet tells my vet  Stops the shysters!


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## ChuckAtBioTracking

If kids have been fed properly heat-treated colostrum and milk from CAE-positive does, there is a good chance they will test positive on an AGID- or ELISA-based test, since the antibodies in the milk will be passed through the digestive tract into the serum of the kids. Their "positive" result will remain positive for a few week after weaning until these antibodies are cleared from the serum. If a kid has become infected and the virus is actively reproducing, these kids (less than a year old) may or may not test positive. In many mammals, the immune system is not developed enough to respond to foreign invaders by producing proper antibodies (one reason why mom's milk is always best! Full of those passive antibodies to protect the little fellers until they can defend themselves). So, if your kids test positive - most likely "false" if they have been given positive milk. If they test negative the jury is still out for the aforementioned reason and one must also consider the possibility of the virus going dormant as well. If you are running mixed herds, or bringing in new animals, my suggestion is to test with PCR at some point to try and rule out dormant virus. As with most tests, it's not 100% accurate either, but it is the only "direct" test in terms of looking for dormant virus. 

Chuck

"The bigger they are, the harder they fall on you" Mark Knopfler


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## LLB101

I found a bred doeling with a lochia type bloody backside tonight. Ugh. Super disappointing, I assume she's lost the pregnancy. I suspect getting head butted as I had to juggle who was in what pens in a lousy rain deluge here one day, after 2 teenage doelings got on the goat's roof and did some untimely damage. I didn't see any bad behavior, so I thought it was ok, but it only takes once. So bummed.

I suppose there's a chance that she didn't lose it, just suffered some damage and salvaged it? Chances are low, I would think. How long should I wait to pull blood to find out? I will want to get her rebred asap for fall freshening, what are the guidelines?


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## ChuckAtBioTracking

It takes about 4 weeks for PSPB to clear from the serum, but we might be able to get an idea of what is going on if you pull a sample ASAP, then again in 10 - 12 days. Send both to me and I can run them on a quantitative assay and determine the relative concentrations of PSPB between the two samples and see if we are looking at a significant change. If she did lose embryos, I would expect to see around a 50% reduction in PSPB levels.

Chuck

"you can always see it coming but you can never stop it" Michael Timmins


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## LLB101

Thanks Chuck, sorry I didn't get back here quickly. I had other issues including computer screen go out and take days to get fixed. In the meantime, her discharge was full on lochia type for a week plus, I can't imagine any portion of the pregnancy survived. I will try to draw blood still if I can get a handle on some other things here.


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