Mar 20, 2024
REDI-NET T-1A ACTIVE VERTEBRATE TICK FIELD SAMPLING
- 1University of Notre Dame
Protocol Citation: REDI-NET Consortium 2024. REDI-NET T-1A ACTIVE VERTEBRATE TICK FIELD SAMPLING. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvw1p5wlmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 18, 2024
Last Modified: March 20, 2024
Protocol Integer ID: 96862
Keywords: Ticks, active collection, vertebrate, iDNA, field collection
Funders Acknowledgement:
USAMRAA
Grant ID: W81XWH-21-C-0001
USAMRAA
Grant ID: W81XWH-22-C-0093
USAMRAA
Grant ID: HT9425-23-C-0059
Disclaimer
This work is supported by the US Army Medical Research and Development Command under Contract No.W81XWH-21-C-0001, W81XWH-22-C-0093 and HT9425-23-C-0059. The views, opinions and/or findings contained in this report are those of the author(s) and should not be construed as an official Department of the Army or Navy position, policy or decision unless so designated by other documentation.
Abstract
To outline steps for properly collecting tick samples actively collected from cattle to evaluate the risk of zoonotic disease transmission by the detection of pathogens from invertebrate DNA (iDNA).
Guidelines
OBJECTIVE
To document the field processes for collecting ticks from non-human vertebrate animals.
SUMMARY/SCOPE
The overarching aim of the REDI-NET is to develop a collaborative laboratory network between domestic and international partnering institutions to address disease surveillance needs in order to effectively detect, predict and contain potentially emergent zoonosis. This SOP provides guidance of the protocol for the correct active tick sampling at CONUS/OCONUS sites to evaluate and predict the risk of zoonotic disease transmission.
RESONSIBLE PERSON
Principal Investigator, Study Coordinator, Entomology Component Lead, Managers
Note
All study procedures must be conducted in compliance with national and local policies for prevention and control of COVID-19 infection.
Materials
| A | B | C | |
| Equipment / Material | Description | Mfg / Product # | |
| Nitrile gloves | Disposal, non-latex gloves | Locally sourced | |
| Rubber boots | Personal protective equipment | Locally sourced | |
| Falcon tubes | 15 ml or 50 ml for sample storage | Locally sourced | |
| Forceps | For sample collection | Locally sourced | |
| Bleach wipes | For material cleaning | Locally sourced | |
| Timer | For sampling timing | Locally sourced | |
| Squeeze chute | For cattle restraint | Locally sourced | |
| Ice/Cooler | For transport of samples | Locally sourced | |
| Tablet | For data entry and picture of sampling site | Locally sourced | |
| Writing utensils | Pencil/pens, marker pens | Locally sourced |
Safety warnings
NOTE: Cattle are restrained in a squeeze chute for all procedures.
- Never stand in front of the chute until the animal has been completely restrained.
- Always be aware of the location of the gate hinge when standing next to the chute as it is opening and closing.
- Always make sure that all drop-down bars and fold-out panels are closed before the chute is opened.
- Always communicate verbally with the team operating the squeeze chute when you are finished checking the animal so that they know when it is safe to open the chute.
- Always be aware of the fact that the animal can still move somewhat inside the chute. Be aware of where your hands and arms are, and be able to remove them from the chute quickly.
- If you ever feel uncomfortable checking an animal (animal is too worked up or aggressive in the chute), or you are unable to safely access part of the animal in the chute, skip it.
Before start
NOTE: Obey all safety procedures in this SOP.
Cattle are restrained in a squeeze chute for all procedures.
- Never stand in front of the chute until the animal has been completely restrained.
- Always be aware of the location of the gate hinge when standing next to the chute as it is opening and closing.
- Always make sure that all drop-down bars and fold-out panels are closed before the chute is opened.
- Always communicate verbally with the team operating the squeeze chute when you are finished checking the animal so that they know when it is safe to open the chute.
- Always be aware of the fact that the animal can still move somewhat inside the chute. Be aware of where your hands and arms are, and be able to remove them from the chute quickly.
- If you ever feel uncomfortable checking an animal (animal is too worked up or aggressive in the chute), or you are unable to safely access part of the animal in the chute, skip it.
SAMPLING TEAMS
SAMPLING TEAMS
This SOP assumes that the animals’ owners/handlers/veterinarians are on site to corral cattle, move them through the chute, and operate the chute. The three minute search time assumes at least three study personnel are available for tick collection. If only two individuals are present, increase search time to five minutes.
One person should be assigned to search the head/neck/dewlap region, one person should be assigned to search the legs/belly region, and one person should be assigned to search under the tail and the udder.
Note
It is suggested not to use the tablet for data collection unless a dedicated collection individual is available among the study personnel as your hands are likely to get very dirty and could contaminate the tablet.
Record the ambient temperature outside the barn or enclosure if working indoors.
SAMPLE COLLECTION
SAMPLE COLLECTION
Once an animal is completely restrained in the squeeze chute, label a collection tube with the date, collection site, and animal’s ear tag number.
Set a timer for three minutes. (See above note in Step 1.)
Lower the rear drop-down bars to access the tail. Lower the lower fold-out panels to access the legs. If necessary, the front swing arm can be used to pin the animal’s head to one side.
Note
These are heavy. Use caution when opening and closing parts of the chute.
Moving your hands over the animal, feel for bumps on the skin. Ticks will be round or flat, smooth, and somewhat moveable, compared to scabs, burs, and warts.
When you feel a tick, grasp it close to the skin at the mouthparts either using your fingers or forceps. Pull forcefully straight out to remove the tick.
Place the tick into the labeled tube. No more than 10 live ticks should be placed in a vial. All stages should be collected. Place a blade of grass in vial before capping. Close the tube to ensure that no ticks escape.
At the end of the three minute search period, close the drop-down bars and fold-out panels and verbally confirm with the chute operator that you are done.
Note
If you are having trouble closing something, make sure you let the operator know not to release the animal until all panels are closed and secured.
For each animal, indicate whether ticks were collected from that individual (‘Yes’), whether the animal was checked, but no ticks were found (‘No Ticks Present’), or whether that animal was not checked for ticks (‘No’) on the data collection sheet.
Repeat this process until at least 20% of the animals have been sampled. Ideally, all animals will be sampled, but occasionally this will not be possible due to time or personnel restraints.
Transfer collection tubes to a cooler on ice for transport back to the lab.
TICK STORAGE
TICK STORAGE
If ticks do not need to be kept alive for research purposes:
Place collection tubes into a -20C freezer for at least five hours, and up to 24 hours to kill ticks.
Ticks should then be stored at -80C until they are identified to species and processed for TNA extraction. (Follow the REDI-NET T-3 Tick Storage.)
If ticks are to be kept alive for research purposes:
Place collection tubes into a refrigerator (4C). It is not recommended that ticks remain in the refrigerator for more than two days prior to processing. (Follow the REDI-NET T-3 Tick Storage.)
Proceed with sample testing following the REDI-NET SOP T-2 Tick Processing.
Protocol references