Table 1.

Clinical data collected, samples tested for Mycoplasma hyopneumoniae (MHP) PCR, and pathologic evaluations throughout the study

Samples	Frequency	Levela	No. of possible samples	No. of collected samples	Testing	Outcome
Coughs
Over 27-min	2 × week	Individual	702	621	Blind observerb	Total coughs per pig (or per group)
Over 24-h	Real-time	Room	355	303	Digital recording systemc	Total coughs per group
Lung lesions
Gross	Necropsy	Individual	39	37	Blind observerb	% lung affected
Microscopic	Necropsy	Individual	39	39	Histopathologyb	Score 0, 1, 2, 3, 4
Tracheal swab	2 × week	Individual	507	489	PCR Protocol 1d	MHP DNA Pos or neg; Ct value
Oral fluid	Daily	Rooms (1–5)	322	322	PCR Protocol 1	MHP DNA Pos or neg; Ct value
Drinker	1 × week	Room (1–5)	45	45	PCR Protocol 1	MHP DNA Pos or neg; Ct value
Air	3 × week	Room (2, 4, and 5)	84	84	PCR Protocol 2e,f	MHP DNA Pos or neg; Ct value

^aRoom 1: One MHP-inoculated pig commingled with 8 uninoculated pigs. Room 2: 3 MHP-inoculated pigs with 6 uninoculated pigs. Room 3: 6 MHP-inoculated pigs with 3 uninoculated pigs. Room 4: 9 MHP-inoculated pigs. Room 5: 3 uninoculated pigs

^bObserver blinded to the MHP inoculation pig status

^cTotal number of coughs per day identified using a commercial digital quantitative cough recording system and software (SoundTalk®, SoundTalks NV, Leuven, Belgium)

^dPCR Protocol 1: TaqMan® Fast Virus 1-Step Master Mix (Life Technologies, Carlsbad, CA USA) with AmpliTaq® 360DNA Polymerase (5U/uL) (Thermo Fisher Scientific, Inc., Waltham, MA USA)

^ePCR Protocol 2: TaqMan® Fast Virus 1-Step Master Mix (Life Technologies)

^fCollected from Room 2, 4, and 5