Nathan Lockwood

Summary Paper

EXPERIMENTAL INFECTION IN LAMBS WITH A RED DEER (CERVUS ELAPHUS)

ISOLATE OF ANAPLASMA PHAGOCYTOPHILUM

By: Snorre Stuen, Wiebke Scharf, Sonja Schauer, Felix Freyburger, Karin Bergstrom,
and Friederike D. von Loewenich

Anaplasma Phagocytophilum, a biological agent transmitted by a certain species

of ticks, was discovered to be the causative agent in a tick borne fever in certain
domestic animals, including sheep and cattle. It was later discovered that the same
TBF was also found in horses, dogs, cats and people. This showed that TBF can be
spread via many animal species.
For many years, TBF has been plaguing norwegian sheep, greatly harming the
sheep industry in Norway, how ever, it is unclear on how the sheep are becoming
infected with ticks. It has been known for a while now that Red Deer in Norway are
important hosts for the vector of A. phagocytophilum.
The study was setup to test a theory that was created that Red Deer were
causing the TBF Infections in sheep. If a connection between Red Deer carriers of Tick
borne TBF and the infections of sheep in Norway were established, then proper actions
could be taken to prevent such infections in sheep.
The study was set up using 16, 5 month old lambs of the dala breed. The lambs
were unrelated and healthy. As a side note, written informed consent was gotten from
the National Animal Research Authority. This consent gave authority to infect some of
the lambs with a specific strain of TBF. After this was done, A. Phagocytophilum was
obtained from a sheep with acute TBF. Red Deer Isolate 7074 was taken from an
apparently healthy Red Deer. 6 Milliliters of heparinized heart blood were inoculated into
one susceptible lamb.

After that, group one of lambs were infected with the ovine isolate on day 0 and
challenged with red deer Isolate on day 42. Group two had the reverse of group one,
and group 3 were infected with red deer isolate on day 60 and challenged with red deer
isolate on day 42.

After the trial period, PCR was performed on DNA from the whole blood sheep.
The DNA was then sequenced and then analyzed. Both the Red Deer isolate and the
lamb isolate showed typical symptoms of TBF, but there were significant differences
between the two, such as incubation time, and fever length. Lambs infected with the
Red Deer strain, and then challenged with the Red Deer strain did not react with clinical
signs, while lambs infected with the Red Deer strain and then infected with the ovine
strain did show clinical signs. Animals inoculated with sheep isolate and then challenged
with the Red Deer strain were fully protected or only subclinically infected. When 497 bp
of the 16S rRNA gene was analysed, the A. Phagocytophilum sheep isolate and the
reed deer isolate 7074 were 100% identical.

Lambs inoculated with a Red Deer isolate of A. phagocytophilum developed

signs of TBF. This means that the red deer might be hosts for ovine infections. A
significant difference in clinical manifestations and infection rate was observed between
the red deer isolate and the ovine isolate. Strains of A. Phagocytophilum differ
somewhat in the severity of the clinical symptoms.

It is unknown whether the particular isolate used from the Red Deer is
representative of the strain cycling around in the wild. It is unclear whether the milder
symptoms of the animals are due to the host adaptations or to normal strain variance. In
this way the study was limited.