Literature Worksheet (10 points)

Name: Allen Koshy TA: Jeffrey Ecklund Section: L04 Date: 7/23/15
1. State the fundamental question, discovery, or nature of the problem(s), question(s), issue(s), or
controversies driving the research efforts in this investigation. (1 point)
The researchers are trying to assess the relationship between this infectious agent
transmitted by ticks of the specific genus Ixodes and Lyme disease and whether or not it may be
the cause for it.
2. Identify and define key terminology that is important in this paper. (1 point)
I.dammani: a spirochete that binds immunoglobulins of patients convalescing from Lyme
disease
Spirochete: a twisted, spiral bacterium associated with syphilis
ECM: erythema chronicum migrans; a skin lesion that may cause cardiac abnormalities,
migratory polyarthritis, intermittent attacks of oligo articular arthritis, or chronic arthritis
in the knees
Hematoxylin +Eosin: a common staining method where hematoxylin colors the nuclei
of cells blue followed a counter staining of eosin which color eosinophilic structures red
Giemsa: a specific stain that helps with the diagnosis of malaria and other parasites.
Stains human cells purple while stains bacterial cells pink
Treponeme: a spirochetes that is parasitic and pathogenic in humans that usually is the
cause for syphilis and yaws
Macules: small changes in the color of the skin, and can sometimes be blemishes in the
skin
Papules: solid elevation of skin with no visible fluid
Indirect immunofluorescence method: uses two antibodies where the primary antibody
is unconjugated and a fluorphore-conjugated secondary antibody directed against the
primary antibody is used for detection.
Antigenically: a mechanism where an infectious agent alters its surface proteins in order
to evade a host immune response.
Suspension: a heterogeneous mixture in which solute-like particles settle out of a
solvent-like phase after being introduced
Etiology: The cause or reason for something
3. List 2 key points made in the paper and describe why they are critical to understanding this
paper. (1 point)
One point is that it takes 10 to 12 weeks after the ticks had engorged where macules and
papules started to appear in the skin of the rabbit and the after 3 to 5 days these lesions had
enlarged. This is important observation made by the paper because it associates a similar
diagnosis for Lyme disease and uses it as a comparison. Another important point is they claimed
that I. dammini spirochete is antigenically related to Lyme disease due to the positive reactions
they obtained by the serum samples. This is important because it suggests a mechanism of how
this bacterium invades the host and how the researchers figured that out through serum samples
of patients that were already diagnosed with Lyme disease.

4. List the methods used to collect the data used in this paper. (1 point)
The researchers isolated the specific bacterium I. dammini and collected from late
September and early October 1981 by flagging lower vegetation on Shelter Island. Spirochetes
that were obtained from ticks were then stained with Giemsa to prepare for microscopy. Electron
miscrosopy was then used of the mid gut diverticula to observe the cells more closely and its fine
structures. Spirochetes were then isolated and inoculated with 0.1 ml of suspension from the
midgut tissues of four infected ticks. After incubation, spirochetes were regularly subcultured
were maintained at 35 degrees Celsius. About 300 of the bacterium were allowed to feed on eight
New Zealand White rabbits where blood smears were examine daily for 14 days. About 10 to 12
weeks after the ticks have placed, observations such an abnormal growths or lesions were noted.
Specimen were then stained hematoxylin and eosin to observe the lesions. Finally, an indirect
immunofluorescence method was used for detection of similarities to Lyme disease patients.
5. Define the major ideas or main hypotheses with corresponding predictions addressed in this
paper. (1 point)
The major ideas that this paper tries to illustrate is that the author is trying to find some
observations that suggest that this specific bacteria strain, I. dammini , is related to the cause of
Lyme disease. The author illustrates points such similar structural components of these
spirochetes and also emphasizes on the observations on growth of those lesions and how they are
enlarged and very similar to patients with Lyme disease.
6. What types of data were collected? Are controls identified in the paper? If so, describe them
briefly. (1 point)
The types of data that were collected very various images obtained through microscopy
of the stained cells during the Gimesa stain as well as the Hematoxylin and Eosin stain. Various
measurements were also taken such the length of the lesions that were observed after the ticks
inhabited in rabbits. A table was organized during the indirect immunofluorescence where it stats
the name of the patient, month and year his/her Lyme disease was contracted and when the
serum was collected as well as Serum dilution end point of each patient. Controls were not
directly mentioned in specific, but it may be assumed that some ticks that placed on the rabbits
did not contain the spirochete bacteria of interest which would show a proper control.
7. Describe 2 results that explain, support, or refute the hypothesis of the paper. Identify key
tables and graphs needed to understand results and conclusions. (1 point)
One result that was very important is that after 10 to 12 weeks, the authors observed
macules and papules that appeared in the skin and how these specific lesions also enlarged after
3 to 5 days. This result supports the hypothesis that this specific bacteria strain may have
something to do with Lyme disease, because the observations show a similar diagnosis to the
disease itself. Another result that supports the authors hypothesis, is that through electron
microscopy of the midgut diverticula, spirochetes were revealed to be closely associated with the
microvillar brush border of the gut epithelium indicated by Figure 2. This was an important
observation and also supports the authors hypothesis because these very similar structural
features were similar to Treponema species which is related to Lyme disease.

8. Describe 2 conclusions the authors discuss in the paper. (1 point)

One conclusion the author discusses in the paper that the treponema-like organism
isolated from I.dammini may be involved in the etiology of Lyme disease. Another conclusion
that the author discusses is that their results show the susceptibility of the rabbit to the specific
bacteria strain and how indirect immunofluorescence test can be used as a possible diagnostic
tool for Lyme disease.
9. Are there additional questions for future research raised by the authors? If so, describe them
briefly. (1 point)
The researchers suggested that additional investigations are needed not only in the
epidemiology and ecology of Lyme disease and related disorders, but also into the relations
between spirochete and I. dammini.
10.State one question that came to mind as you were reading this paper that was not in the paper.
(1 point)
One question that came to mind, is how this indirect immunofluorescence method
specifically support the idea that spirochetes may have something to do with the cause of Lyme
disease? What specific of the results such in Table 1 make this a possibility?