Darin Dutson

Prof. Brandy Conrad

Biology Lab, Friday, 4 P.M.
24 October 2014

West Nile virus cluster analysis and vertical transmission in Culex pipiens complex
mosquitoes in Sacramento and Yolo Counties, California, 2011

Introduction
Researchers and the Sacramento-Yolo Mosquito and Vector Control District (SYMVCD) are
observing and testing to better understand the vertical transmission of West Nile virus (WNV) in
the state of California; particularly in Yolo and Sacramento Counties.
WNV-infested vectors entered the southern part of California in 2003, and by 2004 the virus
was also found in northern parts of the state. Scientists were alarmed at the speed the disease was
being spread. Primary carriers of WNV are genus Culex, comprising of Culex pipiens and Culex
quinquefasciatus. These types of mosquitos vertically transmitted the disease to offspring during
the laying of the eggs.
The researchers decided to do a study on these mosquitos because of the lack of knowledge
in vertical transmission that was taking place with the insects. If more is understood about the
subject, experts could better predict how WNV will spread and what types of insects are most likely
to carry the illness. It is very relevant to the world in that we can prevent serious sickness or even
death if we can understand the patterns of this virus.
Researchers performed the experiment because verification of the aspects that are linked
with high levels of WNV would lead to improvements on controling of the virus. Scientists also
wanted to locate the heart of the dilemma so that they could prevent further spreading of the virus.
By proving that vertical transmission was happening they could begin efforts to stop the disease
from spreading. With this information, many lives could be saved and many cases of serious illness
could be prevented. By understanding these processes, scientists could hinder the spreading of
WNV not only in California, but also in other locations around the globe.
Darin Dutson
Prof. Brandy Conrad
Biology Lab, Friday, 4 P.M.
24 October 2014

Scientists felt vertical transmission of the disease may supplement virus amplification
during summer and provide a mechanism to infect overwintering female mosquitoes during fall
(Fechter-Leggett, et al. 2012). This would mean that the virus would continue in the same areas
when spring arrived, creating a cycle of the disease.
Materials and Methods
The SYMVCD started by placing traps containing a certain food made for swine and rabbits.
They then placed mosquito traps in different areas where there was a detection of WNV. The pools
where WNV tested negative were used as controls. Data were also collected on a seven-day basis,
evading partiality to specific week days. The vertical transmission rate was determined by taking
the percentage of the females that passed the disease to their offspring; they did this by dividing the
total number of mosquitos that passed the disease to offspring by the total of females that were
infected but simply laid eggs without passing disease. They then determined placement of the traps
using a procedure from a space-time scan statistic, also known as SaTScans, which identified
areas and periods where there were many mosquitos that carried WNV (Fechter-Leggett, et al.
2012).
Culex pipiens were kept by being placed into 50 ml containers which were covered with a
type of screen. They placed 5 ml of water so that the mosquitos could lay eggs. The mosquitos were
held at a temperature of twenty-six degrees Celsius. RNA was then extracted using a polymerase
chain reaction with a thermocycler.
Results
Out of just under 1500 sites that were tested, there were 90 sites that tested positive for the
disease gathered from the aforementioned traps. This entailed a rough estimate of about nine
female mosquitos per 1000 were infected with WNV in the counties of Yolo and Sacramento. As for
vertical transmission, out of 297 mosquitos that laid eggs, 11 were found with WNV RNA. If the
Darin Dutson
Prof. Brandy Conrad
Biology Lab, Friday, 4 P.M.
24 October 2014

females were known to already have the virus, there was a fifty percent rate of laid eggs and forty
percent of the larvae from those eggs that were infected with WNV.
Researchers found that because of the certain plaque forming units and the cycle
threshold scores, the approximate minimal filial infection rate (MFIR) was two diseased females
per 1000 (2:1000) for the sites with only one larva per pool that was tested positive for the disease.
For the sites that were entirely infected with WNV, the MFIR was four diseased females per 1000
(4:1000) (Fechter-Leggett, et al. 2012).
Discussion
There remains a lack of information if vertical transmission raises levels of WNV during the
summer months; however, the scientists found that this indeed had an effect on the two northern
counties in California. Through all this, though, the experiment proved their hypothesis correct.
They found that if the virus was vertically transmitted to the offspring, then after winter ended the
virus was active again in the same place during the next season. This means that the virus was
stored in the deposits of eggs that were left over the winter.
Because of the findings with the vertical transmission and the percentage of eggs affected
by the virus, they were able to determine that the virus infected the counties in northern California
during late summer and fall. The researchers determined that the MFIR was probably a more
accurate reading of the rate at which the virus was vertically transferred. Although this is a low
reading, it could have large effects where the mosquitos are great in number.
The sample was very restricted as to the size, and also the amount of time the study was
done. In the future the scientists would like to have more traps set at more locations where WNV is
prevalent. They also would prefer to have the testing be spanned over a greater period of time so
that more predictions and estimations could be made for transmittance of the disease.

Darin Dutson
Prof. Brandy Conrad
Biology Lab, Friday, 4 P.M.
24 October 2014

Bibliography
Fechter-Leggett, Ethan, Nelms, Brittany M., Barker, Christopher M., and Reisen, William K. "West Nile
Virus Cluster Analysis and Vertical Transmission in Culex Pipiens Complex Mosquitoes in
Sacramento and Yolo Counties, California, 2011." Journal of Vector Ecology No. 2 37 (2012): 442-
49. Web. 24 Oct. 2014.