Table 3.

. Summary of results obtained in the standard (ST) and high bioactivation (HB) crosses of the Drosophila wing SMART after scoring marker-heterozygous flies (mwh +/+ flr³, wild-type wings) treated with nicotine (NIC) at [0, 0.0125, 0.025, 0.05, 0.456, 1.9, 3.8, 11.4 mM]; urethane (URE)[20 mM] and Milli-Q water as positive and dissolvent controls, respectively.

Compound Crossb		Number of flies	Spots per Fly (Number of Spots) Statistical Diagnosisa					Mean	Clone formation
	Type		Small single spots	large single spots	Twin spots	Total spots	mwhclones	mvh clone size class	per 105 per cell	cells divisionc
			(1-2 cells)	(>2 cells)					observed	control connected
			m = 2	m = 5	m = 5	m = 5
ST
Positive control
Water		61	0.46 (28)	0.07 (04)	0.02 (1)	0.54 (33)	33	1.70	1.10
URE	20.0	28	1.21 (34) +	0.11 (03) -	0.04 (1) -	1.36 (38) +	38	1.45	2.80	1.65
NIC treatments
Water	0	61	0.46 (28)	0.07 (04)	0.02 (1)	0.54 (33)	33	1.70	1.10
NIC	0.0125	44	1.16 (51) +	0.43 (19) -	0.00 (0) -	1.59 (70) +	70	2.26	3.25	2.15
NIC	0.0250	41	0.66 (27) -	0.00 (00) -	0.02 (1) -	0.68 (28) -	28	1.21	1.40	0.30
NIC	0.0500	21	0.67 (14) -	0.10 (02) -	0.05 (1) -	0.81 (17) -	17	1.47	1.65	0.55
Water	0	51	0.47 (24) -	0.10 (05)	0.00 (0)	0.57 (29)	29	1.69	1.15
NIC	0.4560	38	0.68 (26) -	0.05 (02) -	0.00 (0) -	0.74 (28) -	28	1.54	1.50	0.35
NIC	1.9000	46	0.57 (26) -	0.04 (02) -	0.02 (1) -	0.63 (29) -	28	1.57	1.25	0.10
NIC	3.8000	32	0.62 (20) -	0.03 (01) -	0.03 (1) -	0.69 (22) -	22	1.45	1.40	0.25
NIC	11.4000	22	1.27 (28) +	0.23 (05) -	0.05 (1) -	1.55 (34) +	34	1.71	3.15	2.00
HB
Positive control
Water	0	61	0.66 (40)	0.07 (04)	0.00 (0)	0.72 (44)	44	1.43	1.50
URE	20.0	23	2.52 (58) +	0.39 (09) +	0.17 (4) +	3.09 (71) +	71	1.82	6.35	4.85
NIC treatments
Water	0	61	0.66 (40)	0.07 (04)	0.00 (0)	0.72 (44)	44	1.43	1.50
NIC	0.0125	60	0.68 (41) -	0.05 (03) -	0.02 (1) -	0.75 (45) -	42	1.14	1.45	-0.05
NIC	0.0250	60	0.65 (39) -	0.03 (02) -	0.00 (0) -	0.68 (41) -	41	1.29	1.40	-0.10
NIC	0.0500	60	0.77 (46) -	0.08 (05) -	0.00 (0) -	0.85 (51) -	51	1.61	1.75	0.25
#1 Assay∗
Water	0	35	0.54 (19)	0.06 (02)	0.00 (0)	0.60 (21)	21	1.29	1.25
NIC	0.4560	99	0.38 (38) -	0.07 (07) -	0.03 (3) -	0.48 (48) -	47	1.64	0.95	-0.25
NIC	1.9000	58	0.47 (27) -	0.05 (03) -	0.07 (4) -	0.59 (34) -	34	1.82	1.20	-0.05
NIC	3.8000	69	0.36 (25) -	0.06 (04) -	0.00 (0) -	0.42 (29) -	28	1.61	0.85	-0.40
#2 Assay∗
Water	0	50	0.50 (25) -	0.10 (05)	0.04 (2)	0.64 (32)	32	1.66	1.30
NIC	0.4560	60	0.33 (20) -	0.08 (05) -	0.02 (1) -	0.43 (26) -	25	1.64	0.85	-0.45
NIC	1.9000	58	0.47 (27) -	0.05 (03) -	0.07 (4) -	0.59 (34) -	34	1.82	1.20	-0.10
NIC	3.8000	60	0.37 (22) -	0.05 (03) -	0.00 (0) -	0.42 (25) -	24	1.42	0.80	-0.50

^aStatistical diagnoses according to Frei and Würgler (1988) m: minimal risk multiplication factor for the assessment of negative results. For the final statistical diagnosis of all positive (+) and negative (-) results; the non-parametric Mann-Whitney and Wilcoxon U-test with significance levels α and β = 0.05 was used to exclude false positive or negative diagnoses [Frei and Würgler, 1995]. One side binomial test, significance levels α and β: significant results: + (α ≤ 0.05; no significant results: - (β ≤ 0.05).

^bST: standard cross; HB: high bioactivation cross.

^cClone frequencies per fly divided by the number of cells examined per fly (48,800) gives an estimate of formation frequencies per cell and per cell division in chronic exposure experiments [Frei and Würgler, 1995].∗In HB cross the NIC [11.4 mM] was lethal.