Hello,
I am planning a reserch which involves analysis of saliva cortisol in guinea pigs. Unfortunately, I am currently having trouble collecting saliva samples from my guinea pigs. I am not able to collect enough sample for ELISA analysis. I found publications where researches collected samples by inserting standard cotton bud into the animals' cheek pouches for 30 seconds and obtained enough sample for analyses. I have also found several publications indicating that using cotton swabs for sample collection may interfere with the results. I tried to use both Salimetrics' SalivaBio Children's Swab and standard cotton swabs, I inserted them in guinea pig mouth for approximately 3-5 minutes then centrifuged them for 15 min at 5000 rpm however I still didn't obtain more than 25 µL of sample. Moreover, the sample I obtain is not clear but brownish as it is contaminated with the food the guinea pigs have been eating prior to sampling. Also, some of my guinea pigs refused to keep the swabs in place and tried to remove it from their mouth or started chewing on it so it made sampling harder.
I would be grateful if someone experinced with this type of research could share their insight about some of the issues I am experiencing:
- Does using cotton swabs, in your experience, has any effect on the results of the cortisol analysis?
- How do you get guinea pigs to not resist inserting swabs into their cheek pouches and stop them from chewing them?
- Do you irrigate guinea pig mouths before sample collection to obtain saliva uncontaminated by food or perhaps withdraw food from animals prior to sampling? In your experince how much does the contamination of sample with food affect the results?
- How much sample do you gain by your method and do the time and speed of centrifugation have significant effect on the volume of sample you obtain?
Thenk you and kind regards,
Anja
I have no experience in this area. I would think that having the animals hungry and then placing food just out of reach, would cause them to salivate. Whether it is enough to gather more saliva I don't know.
When humans get tested for cortisol it is usually first thing in the morning after fasting.
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