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MB Sample ID: SA086836

Local Sample ID:19-4N-H24
Subject ID:SU001291
Subject Type:Mammal
Subject Species:Sus scrofa
Taxonomy ID:9823
Age Or Age Range:2 days after birth
Weight Or Weight Range:above 1.3Kg at birth
Gender:Female
Animal Animal Supplier:Purdue University Animal Sciences Research and Education Swine facility
Animal Housing:Education Swine facility
Animal Water:ad libitum
Animal Inclusion Criteria:Three to four gilts above 1.3Kg at birth were selected per litter from eight different sows which were monitored during parturition

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Subject:

Subject ID:SU001291
Subject Type:Mammal
Subject Species:Sus scrofa
Taxonomy ID:9823
Age Or Age Range:2 days after birth
Weight Or Weight Range:above 1.3Kg at birth
Gender:Female
Animal Animal Supplier:Purdue University Animal Sciences Research and Education Swine facility
Animal Housing:Education Swine facility
Animal Water:ad libitum
Animal Inclusion Criteria:Three to four gilts above 1.3Kg at birth were selected per litter from eight different sows which were monitored during parturition

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
19-4N-H24SA086836FL012873MTreatment
19-4N-H24SA086836FL012873milkSample Type

Collection:

Collection ID:CO001285
Collection Summary:Blood samples were collected from gilts at 48 h postnatal. Gilts were euthanized approximately 48 h after birth using CO2 inhalation. Following euthanasia, skin of the abdominal and genital regions was cleaned thoroughly using 70% ethanol, and vaginal swabs were taken using a cytology brush (Puritan 2196 Removable Stiff Bristle Tip Brush; QuickMedical; Issaquah, WA) by inserting the tip of the brush into the vulva angled dorsally at 45˚. Once inserted to the base of the bristles, the brush was rotated 360˚ against the vaginal surface. Two consecutive swabs were collected from each animal, and swabs were placed in separate 15 ml sterile polypropylene conical tubes (Falcon™, Fisher Scientific, San Jose, California) and immediately placed on dry ice for transport. Samples were stored in a -80°C freezer until lipid extraction and analysis. Sows were milked during farrowing, and at 24 h after delivery of first piglet. For milk collection piglets were removed from the sow for approximately an hour and then 1 ml oxytocin (VetOne; Boise, ID; 20 USP/ml) was administered IM using a 20g x 1.5-inch needle into the vulva to stimulate milk letdown. Colostrum samples were collected manually from all teats and combined to create a uniform sample. Samples were stored until further analysis at -20°C
Sample Type:Vaginal epithelium
Collection Method:swab
Collection Location:West Lafayette
Collection Frequency:once
Collection Duration:1min
Storage Conditions:-80℃
Collection Vials:Puritan 2196 Removable Stiff Bristle Tip Brush; QuickMedical; Issaquah, WA
Storage Vials:15 ml sterile polypropylene conical tubes (Falcon™, Fisher Scientific, San Jose, California)
Additives:none

Treatment:

Treatment ID:TR001306
Treatment Summary:Three to four gilts were selected per litter from eight different sows which were monitored during parturition. Immediately after delivery, all gilts were towel-dried, weighed, and placed in a holding cart until at least three gilts above 1.3 kg were delivered. Within litter, each gilt was randomly assigned to one of four treatment groups and ear tagged for identification. The four treatment groups were: suckled (S; n=8); suckled plus fat-supplement (SF; n=5); bottle-fed with milk-replacer (B; n=8), and; bottle-fed milk replacer plus fat-supplement (BF; n=7). Body weights were recorded at birth and 48 h. All gilts were administered a 2 ml dose of Camas experimental antibody product (Camas Incorporated; Le Center, MN) using Pump It™ Automatic Delivery System (Genesis Industries, Inc.) at birth, and at 3 h and 9 h after the first dose.

Sample Preparation:

Sampleprep ID:SP001299
Sampleprep Summary:The Bligh & Dyer lipid extraction technique was slightly modified to extract lipids from the vaginal swab samples. A volume of 500 µl distilled water was added to the conical tube containing the swab brushes and vortexed to remove biological material from the brush. The brushes were removed, the sample homogenate was transferred to a new tube, and phase separation was performed by mixing with chloroform/methanol/distilled water (1:2:0.8). Samples were centrifuged, the organic phase (bottom phase) was separated from aqueous phase, divided into four aliquots, and dried in a centrifugal evaporator (Savant SpeedVac AES2010, ThermoFisher Scientific, San Jose, CA). Dried lipid extracts were stored at -20˚C until mass spectrometry analysis. The Bligh & Dyer lipid extraction technique was also used to extract lipids from 48 h serum samples from suckled and bottle-fed piglets; colostrum samples taken from all eight sows (during parturition, 6 h after first piglet born, 12 h after, and 24 h after); and milk replacer used for bottle-feeding. The procedure for these samples began at the phase separation step (i.e. no water was added to the samples).
Processing Storage Conditions:Room temperature
Extraction Method:Bligh & Dyer
Extract Storage:-80℃
Sample Resuspension:acetonitrile/methanol/ammonium acetate 300 mM at 3:6.65:0.35 (v/v)
Sample Derivatization:none
Sample Spiking:none

Combined analysis:

Analysis ID AN002037
Analysis type MS
Chromatography type Unspecified
Chromatography system Agilent 6410
Column none
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6410 QQQ
Ion Mode UNSPECIFIED
Units ion counts

Chromatography:

Chromatography ID:CH001477
Chromatography Summary:Direct infusion
Instrument Name:Agilent 6410
Column Name:none
Chromatography Type:Unspecified

MS:

MS ID:MS001889
Analysis ID:AN002037
Instrument Name:Agilent 6410 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Pooled sample injections (8 µl) were delivered to the micro-autosampler (G1377A) in a QQQ6410 triple quadrupole mass spectrometer (Agilent Technologies, San Jose, CA) equipped with an ESI ion source. The solvent pumped between injections (CapPump G1376A, Agilent Technologies, San Jose, CA) was acetonitrile with 1% formic acid at 10µL/min. Between sample injections, methanol/chloroform was injected to remove any remaining lipids before the next sample injection
Ion Mode:UNSPECIFIED
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