Table 3.

Statistical results for normalized cycle threshold (C_T) transcription values and changes in normalized C_T transcription values (ΔC_T) for 11 genes of interest during the 4th sampling event (222 days post experiment)

	Statistical approach	Gene	Group CEIR		Group CEI		Group CER		Group CE		Group EIR		Group ER		Group IR
			χ 2	p	χ 2	p	χ 2	p	χ 2	p	χ 2	p	χ 2	p	χ 2	p
CT values	Univariate	AHR	8.95	0.03**	9.03	0.01**	7.80	0.02**	6.81	<0.01**	0.39	0.82	0.01	0.91	0.52	0.47
		ATF	10.56	0.01**	1.97	0.37	5.14	0.08	0.53	0.46	11.80	<0.01**	6.40	0.01**	8.48	<0.01**
		CaM	2.55	0.47	2.02	0.36	1.50	0.47	0.88	0.35	1.17	0.56	0.54	0.46	0.83	0.36
		CD9	5.07	0.17	2.35	0.31	2.95	0.23	1.84	0.17	4.58	0.10	2.50	0.11	3.31	0.07
		CL	5.07	0.17	3.08	0.21	2.78	0.25	1.84	0.17	3.76	0.15	1.60	0.21	2.88	0.09
		HSP70	5.73	0.13	4.49	0.11	5.24	0.07	3.15	0.08	1.02	0.60	0.90	0.34	1.47	0.23
		Lep	7.52	0.06	9.00	0.01**	6.20	0.05**	6.82	<0.01**	0.33	0.85	0.28	0.60	0.13	0.72
		MyD88	5.02	0.17	4.19	0.12	4.02	0.13	3.56	0.06	1.59	0.45	<0.01	0.96	1.19	0.28
		Mx1	8.87	0.03**	3.56	0.17	7.16	0.03**	3.15	0.08	3.46	0.18	0.28	0.60	0.06	0.81
		SAA	7.29	0.06	4.04	0.13	6.40	0.04**	3.15	0.08	1.70	0.43	0.40	0.53	1.19	0.28
		SOD	8.32	0.04**	3.24	0.20	5.34	0.07	2.45	0.11	8.09	0.02**	5.38	0.02 **	4.78	0.03**
	Multi‐variate	All genes	F 3,25 = 2.48, p = 0.03**		F 2,13 = 0.16, p = 0.16		F 2,21 = 2.74, p = 0.04**		F 1,9 = 2.10, p = 0.14		F 2,20 = 2.53, p = 0.04**		F 1,16 = 1.97, p = 0.14		F 1,16 = 3.13, p = 0.03**
Changes in CT values (ΔCT)	Univariate	AHR	3.76	0.29	1.72	0.42	2.89	0.24	1.50	0.22	3.93	0.14	2.88	0.09	1.70	0.19
		ATF	10.85	0.01**	3.33	0.19	6.83	0.03**	0.96	0.33	9.81	<0.01**	5.52	0.02**	6.15	0.01**
		CaM	5.92	0.12	3.61	0.16	2.34	0.31	2.16	0.14	5.38	0.07	1.70	0.19	4.36	0.04**
		CD9	5.62	0.13	4.33	0.11	3.00	0.22	2.94	0.09	3.80	0.15	1.70	0.19	2.89	0.09
		CL	6.51	0.08	3.12	0.21	3.90	0.14	2.16	0.14	5.77	0.06	3.34	0.07	3.82	0.05**
		HSP70	6.64	0.08	2.20	0.33	2.44	0.30	0.00	1.00	7.57	0.03**	2.88	0.09	5.52	0.02**
		Lep	7.75	0.05**	7.10	0.03**	6.37	0.04**	6.00	0.01**	5.40	0.07	4.36	0.04**	1.38	0.24
		MyD88	5.49	0.14	2.40	0.30	3.72	0.16	1.50	0.22	5.74	0.06	3.84	0.05**	2.88	0.09
		Mx1	1.57	0.67	0.91	0.63	<0.01	1.00	0.00	1.00	1.96	0.38	0.00	1.00	1.70	0.19
		SAA	1.21	0.75	0.91	0.63	0.44	0.80	0.00	1.00	1.61	0.45	0.83	0.36	0.43	0.51
		SOD	10.29	0.01**	4.73	0.09	6.02	0.05**	2.94	0.09	9.52	<0.01**	5.52	0.02**	6.15	0.01**
	Multi‐variate	All genes	F 3,23 = 1.80, p = 0.10		F 2,12 = 0.90, p = 0.61		F 2,19 = 1.34, p = 0.26		F 1,8 = 1.06, p = 0.44		F 2,18 = 1.96, p = 0.09		F 1,14 = 3.42, p = 0.05**		F 1,14 = 3.31, p = 0.02**

Differences among treatment groups including Control (C), Exposed (E), Infected (I), and Reference (R) tortoises were evaluated using a nonparametric rank sums tests (Kruskal‐Wallis) using χ ² approximation to assess differences in the means. Transcription profiles were also evaluated using a multivariate approach via permutational multivariate analysis of variation (perMANOVA). Tortoises include five control, five exposed, and five infected captive animals and 14 wild reference tortoises.

^**Significance at α ≤ 0.05.