Allie (Allison) Lloyd

Linda Rittenhouse

Biology 1615-Tuesday 1:00 PM

Final draft

When it comes to the history of the B. Abortus (Brucella Abortus) it has been a very

common disease found in domestic and wild animals. The main species that they find the

bacteria B. Abortus in are bison, cattle, moose, and elk. One important thing to understand about

B. Abortus is the side effects that the researchers already know to be true. Some of the main side

effects are the women cattle having abortions; production of milk becomes decreased, they

become infertile, and for the male cattle they get inflammation in the groin area and this can

cause them to become infertile as well. The scientists already know that B. Abortus in our

animals have been an issue for a while. It was known to be one of the most significant diseases

for cattle around the time period of the 1930s. The researchers chose to study more deeply on B.

Abortus because they wanted to get a better understanding as to why all of these livestock

animals were reacting positively to these bacteria. They were setting out to find the best test that

would help show what was creating the cross reaction between the livestock (elk) and B.

Abortus. In the article the scientist state their hypothesis as, We hypothesized that the western

blot would have greater specificity, while maintaining the sensitivity of standard serology, and

would be a valid and useful confirmatory test for diagnosing B. Abortus infections in elk. (1)

Therefore, they predict that the western blot test will be the most effective test in showing the

cross reaction between the elk and B. abortus bacteria.

As the scientist began looking back at previous tests like the False-Positive test and

serologic test, they soon came to realize their importance. These tests in the past have helped
show positive results of B. Abortus, but they were mainly for cattle herds only. When the

scientist would use the test on wildlife herds the result wouldnt come back normal. From these

tests they found that the bacteria Yersinia Enterocolitica (Y.E) can cause cross reactions in B.

Abortus. This would then come back as a positive test for B. Abortus. The scientist then knew

they needed a better test to help show more accurate results. This is when the scientists began

their process. They started off by gathering four herds of elk. Two came herds came from

Wyoming which contained 28 elk and the other two herds came from California which contained

26 elk. The herds from Wyoming naturally had B. Abortus in their systems and they were all

tested by at least three test coming back with positive results. The California herds were not

known to have come in any contact with the bacteria B. Abortus. After this they then used a

random number generator to randomize the elk. By doing this they no longer knew which elk

came from which herd. The next step was the immunoblot procedure.

An immunoblot procedure is where it allows the antigens to be separated from a cultured

area as they then stick fastly to the sheets. This allows the antigens to bind to the sheets which

then leaves a dye. From this dye, it can help you label each antigen that was tested. The scientist

had created an immunoblots for two types of B. Abortus (Brucella 2308 and RB51), two types of

Y.E (O:8, O:9), and cell lysate. Within these immunoblot there were five lanes, one for each

antigen. All of these were tested twice to make sure the results came back the same. If something

didnt match up they would have ran a third test. B. Abortus, and Y.E O:9 created a non-detached

pattern of proteins that displayed coloring and banding. Therefore, it showed up positive. The

2308 lane showed negative samples when it came to the dye. From the 24 negative results

created 16 came back showing some sign of similarity to Y.E O:9.

 They also used the Standard Brucella (B. Abortus) agglutination testing. In this test, they

take samples that had been evaluated already (they mainly use blood) and once they have the

sample they can run it through a screen that can help detect diseases like B. Abortus. These were

talked about as the serological essays. These results were then compared to the immunoblots. By

doing this it helps take away some of the concern of B. Abortus positive and negative guessed

results. If any result came back as a positive hit for B. Abortus, it would go through more intense

testing.

From this experiment the scientists were able to show the comparison of sensitivity (Se)

and specificity (Sp) from the western blot. The results had shown positive with 100% in Se and

Sp. One thing that was surprising was 5 immunoblots out of 133 immunoblots showed that you

couldn't tell if the elk had a positive reaction to B. Abortus by just looking at them. From this

result the scientist categorized them as an intermediate group. Overall every one of the elk from

the standard (serologic) tests came back positive showing a reaction to Y.E and a negative

reaction to B. Abortus.

From this information, the scientists gathered they easily supported their hypothesis.

They showed how the positive B. Abortus compared to the color staining with the positive

correlation. The results also showed from taking more than one test that it proves the negative elk

came back negative even though they came from a group that was supposedly positive for B.

Abortus. Considering the results the scientists were able to show which tests were more effective

and which ones were not so effective. They showed that the western immunoblot test was the

most effective when it comes to confirming the findings B. Abortus because it is the most

specific with their results and it helps counteract mistakes. The only issue is that it was easier and

cheaper to do the screenings on the elk through the standard serology tests. These tests like the
rivanol and card test from serology testing would be the cheapest and easiest for basic tests and

results of B. Abortus.

From the limited amount of resources of elk, the scientists were able to get their hands

on, they did mention there should be more testing on free ranged elk and other free ranged

species. This would allow them to explore the method more and confirm more cases of B.

Abortus. By allowing them to study a bigger population it would help them to see more of a

pattern on which elk or other cattle would show positive or negative correlations of B. Abortus.

In this experiment the scientist only tested elk. If they were to tests all types of cattle like moose,

bison, elk, and cows they could compare them and see which one is more acceptable to having or

carrying the bacteria B. Abortus.

 Works Cited

Schumaker, B. A., Mazet, J. A., Gonzales, B. J., Elzer, P. H., Hietala, S. K., & Ziccardi, M. H.
(2010). Evaluation Of The Western Immunoblot As A Detection Method For Brucella Abortus
Exposure In Elk. Journal of Wildlife Diseases, 46(1), 87-94. doi:10.7589/0090-3558-46.1.87