Ticks will be collected from areas in the five counties in the state where Lyme disease and other tick-borne infections are prevalent. Trapping sites will be set in known tick habitats in the five counties to help in trapping and collecting small animals, including the target pests. Sherman live-traps baited with oatmeal and peanut-butter will be set at the sites to ensnare the ticks. All captured animals will be regularly and carefully examined to allow the researchers to extract as many ticks as possible. The trapping of animals and extraction of ticks from the captured animals will take about four weeks.
All ticks collected will be placed in vials containing 75% ethanol and sent to parasitology laboratories. The screening process will be done through MasTag polymerase chain reaction (PCR), a multiplication method, which is known for its fast and economic screening of tick-borne pathogens. PCR is also known for efficient and accurate screening of ticks for polymicrobial contagion. Before beginning the PCR procedure, the ticks will be removed from the vials, washed with saline containing phosphate buffer, and then homogenized with Tri-reagent LS.
The next step will involve nucleic acid extraction, where total RNA will be suspended in about 30ml of H2O. The generation of cDNA will then follow where 20ml of the solution and 15ml of total RNA will be reacted using Superscript 11 Reverse Transcriptase as a catalyst. The MassTag PCR procedure will then be done using the tick panel, with primers added to the reactants to help detect the Borrelia species responsible for relapsing fever and the Powassan virus. All the MassTag PCR assays that will be done in the experiment will utilize 3ml of cDNA. The final PCR products will be size-fractioned using ethidium bromide that is stained with agarose gel for efficient sequencing.
The biological nature of ticks is considered an important element when studying the role they play in different tick-borne diseases and how the infections can be treated and most importantly, prevented (Williams, Davidson, Lutz, and Applegate 955). Ticks are scientifically referred to as arthropods and found in the class Arachnida where spiders, mites, and scorpions also fall. Only two out of the three families of ticks have been proved to be of any medical significance (Berrada, Geothert, Cunningham, and Telford 463). There are two major groups of the pest: soft ticks, which fall under the family Argasidae, and hard ticks that belong to the family Ixodidae. The main difference between the two families is the scutum (hard plate) found in hard ticks and absent in their soft counterparts.
To screen and compare ticks based on vegetation and landscape, it is important to understand the role they play in transmitting pathogens to humans and animals. For example, polymicrobial tick infections mostly involve the transmission of a number of pathogens through just a single bite (Koch and Burg 174). Ixodes scapularis is the...