PMID- 29369729
OWN - NLM
STAT- MEDLINE
DCOM- 20181003
LR  - 20181004
IS  - 1943-3700 (Electronic)
IS  - 0090-3558 (Linking)
VI  - 54
IP  - 2
DP  - 2018 Apr
TI  - DEVELOPMENT AND VALIDATION OF QUANTITATIVE PCR ASSAYS TO MEASURE CYTOKINE 
      TRANSCRIPT LEVELS IN THE FLORIDA MANATEE ( TRICHECHUS MANATUS LATIROSTRIS).
PG  - 283-294
LID - 10.7589/2017-06-139 [doi]
AB  - Cytokines have important roles in the mammalian response to viral and bacterial 
      infections, trauma, and wound healing. Because of early cytokine production after 
      physiologic stresses, the regulation of messenger RNA (mRNA) transcripts can be 
      used to assess immunologic responses before changes in protein production. To 
      detect and assess early immune changes in endangered Florida manatees ( 
      Trichechus manatus latirostris), we developed and validated a panel of 
      quantitative PCR assays to measure mRNA transcription levels for the cytokines 
      interferon (IFN)-gamma; interleukin (IL)-2, -6, and -10; tumor necrosis factor-alpha; and 
      the housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 
      beta-actin (reference genes). Assays were successfully validated using blood samples 
      from free-ranging, apparently healthy manatees from the east and west coasts of 
      central Florida, US. No cytokine or housekeeping gene transcription levels were 
      significantly different among age classes or sexes. However, the transcription 
      levels for GAPDH, IL-2, IL-6, and IFN-gamma were significantly higher ( P<0.05) in 
      manatees from the east coast of Florida than they were from those from the west 
      coast. We found IL-10 and beta-actin to be consistent between sites and identified 
      beta-actin as a good candidate for use as a reference gene in future studies. Our 
      assays can aid in the investigation of manatee immune response to physical trauma 
      and novel or ongoing environmental stressors.
FAU - Ferrante, Jason A
AU  - Ferrante JA
AD  - 1   Wetland and Aquatic Research Center, US Geological Survey, 7920 NW 71st 
      Street, Gainesville, Florida 32653, USA.
FAU - Hunter, Margaret E
AU  - Hunter ME
AD  - 1   Wetland and Aquatic Research Center, US Geological Survey, 7920 NW 71st 
      Street, Gainesville, Florida 32653, USA.
FAU - Wellehan, James F X
AU  - Wellehan JFX
AD  - 2   Department of Comparative, Diagnostic, and Population Medicine, College of 
      Veterinary Medicine, University of Florida, 2015 SW 16th Avenue, Gainesville, 
      Florida 32608, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20180125
PL  - United States
TA  - J Wildl Dis
JT  - Journal of wildlife diseases
JID - 0244160
RN  - 0 (Cytokines)
SB  - IM
MH  - Animals
MH  - Cytokines/*blood
MH  - Polymerase Chain Reaction/methods/*veterinary
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Trichechus manatus/*blood
OTO - NOTNLM
OT  - Cytokine
OT  - Trichechus manatus latirostris
OT  - endangered species
OT  - health assessment
OT  - immune function
OT  - manatee
EDAT- 2018/01/26 06:00
MHDA- 2018/10/04 06:00
CRDT- 2018/01/26 06:00
PHST- 2018/01/26 06:00 [pubmed]
PHST- 2018/10/04 06:00 [medline]
PHST- 2018/01/26 06:00 [entrez]
AID - 10.7589/2017-06-139 [doi]
PST - ppublish
SO  - J Wildl Dis. 2018 Apr;54(2):283-294. doi: 10.7589/2017-06-139. Epub 2018 Jan 25.