Broiler intestine DMT1 gene expression and bone characteristics, as affected by in ovo injection of different forms of manganese

Figures & data

Table 1. Specific primer sequences for Real-time PCR.

Gene	NCBI reference	Primer sequence	PCR product length	Annealing temperature
β-actin	NM_205518.1	Forward 5´-GTCCACCTTCCAGCAGATGT-3´	169	63.6
		Reverse 5´-ATAAAGCCATGCCAATCTCG-3´
DMT1	NM_001128102.1	Forward 5´-CCATCCTCAGCGTCATCTACC-3´	191	61
		Reverse 5´-CACCTCCATCTCCCACCGTCA-3´

Table 2. Effect of in ovo injection of different sources of manganese on DMT1 mRNA Expression in different intestinal segments on embryonic day 18 and post-hatch days 1 and 7*.

	E18			P1			P7
Treatments	D	J	I	D	J	I	D	J	I
NC	1.000^b	1.000^b	1.000^b	1.000^bc	1.000^cd	1.000^ab	1.000^c	1.000^a	1.000^a
PC	0.523^b	1.008^b	0.528^cd	0.706^d	0.615^d	0.342^b	0.346^d	0.111^c	0.111^c
MnS	1.180^b	2.209^ab	0.718^c	0.834^cd	1.597^bc	0.403^ab	0.643^d	0.173^c	0.173^c
Mn-Met	0.585^b	1.168^b	0.497^d	1.099^bc	2.112^ab	0.942^ab	1.051^c	0.508^b	0.508^b
NMn	1.718^b	2.848^a	1.504^a	1.434^a	2.252^ab	0.842^ab	2.008^a	0.068^c	0.068^c
NMn-Met	5.545^a	3.338^a	1.437^a	1.237^ab	2.962^a	1.217^a	1.432^b	0.178^c	0.178^c
SEM	0.465	0.243	0.099	0.062	0.203	0.098	0.132	0.083	0.083
p-value	.0003	.0001	.0001	.0001	.0001	.027	.0001	.0001	.0001

*Means within a column with different superscripts are significantly different (p < .05).

NC: Non-injected; PC: Diluent-injected controls; MnS: Manganese sulphate; Mn-Met: Manganese-methionine; NMn: nano manganese; NMn-Met: nano manganese–methionine; D: duodenum; J: jejunum; I: ileum; E18: embryonic day 18; P1: post-hatch days 1; P7: post-hatch days 7.

Table 3. Effect of injection of different sources of manganese on bone length (cm), fresh weight (gr) and dry weight (gr) on embryonic day 18 and post-hatch days 1, 7, 14*.

	E18			P1			P7			P14
Treatments	Length (cm)	Fresh (gr)	Dry (gr)	Length (cm)	Fresh (gr)	Dry (gr)	Length (cm)	Fresh (gr)	Dry (gr)	Length (cm)	Fresh (gr)	Dry (gr)
NC	2.345	0.209^ab	0.050^ab	3.104	0.388	0.130	4.431^ab	0.998	0.316^ab	5.110	3.007^ab	0.978
PC	2.297	0.197^b	0.049^b	3.101	0.380	0.128	4.282^b	0.971	0.333^ab	5.320	2.920^b	1.011
MnS	2.325	0.203^ab	0.049^b	3.116	0.390	0.130	4.411^ab	0.983	0.336^ab	5.115	3.203^ab	1.100
Mn-Met	2.397	0.220^ab	0.054^ab	3.116	0.408	0.133	4.354^ab	1.033	0.308^b	5.314	3.124^ab	1.072
NMn	2.399	0.208^ab	0.051^ab	3.182	0.425	0.132	4.450^a	1.100	0.347^a	5.340	3.641^a	1.098
NMn-Met	2.483	0.224^a	0.056^a	3.129	0.417	0.131	4.396^ab	1.014	0.329^ab	5.203	3.055^ab	1.024
SEM	0.022	0.003	0.0007	0.020	0.008	0.002	0.016	0.021	0.004	0.053	0.073	0.022
p-value	.192	.033	.016	.888	.469	.959	.025	.536	.044	.696	.042	.520

*Means within a column with different superscripts are significantly different (p < .05).

NC: Non-injected; PC: Diluent-injected controls; MnS: Manganese sulphate; Mn-Met: Manganese-methionine; NMn: nano manganese; NMn-Met: nano manganese–methionine; E18: embryonic day 18; P1: post-hatch days 1; P7: post-hatch days 7; P14: post-hatch days 14.

Table 4. Effect of injection of different sources of manganese on percentage of fresh and dry tibia weights and bone ash on embryonic day 18 and post-hatch days 1, 7, 14*.

	E18			P1			P7			P14
Treatments	Fresh (%)	Dry (%)	Ash (%)	Fresh (%)	Dry (%)	Ash (%)	Fresh (%)	Dry (%)	Ash (%)	Fresh (%)	Dry (%)	Ash (%)
NC	0.688^a	0.166^ab	20.924	0.800	0.267	24.956	0.825	0.262	27.455^b	0.801^b	0.260^b	33.526
PC	0.564^b	0.140^b	21.226	0.804	0.272	24.745	0.803	0.276	31.505^ab	0.847^ab	0.292^ab	33.867
MnS	0.636^ab	0.153^ab	22.654	0.855	0.286	24.788	0.816	0.279	31.319^ab	0.957^ab	0.328^a	33.886
Mn-Met	0.710^a	0.174^a	21.092	0.815	0.268	24.975	0.865	0.258	30.713^ab	0.883^ab	0.304^ab	34.613
NMn	0.614^ab	0.150^ab	21.184	0.925	0.288	24.521	0.811	0.259	32.497^a	0.966^a	0.291^ab	36.409
NMn-Met	0.671^ab	0.167^ab	23.928	0.852	0.267	24.811	0.819	0.268	32.072^a	0.860^ab	0.289^ab	35.661
SEM	0.013	0.003	0.952	0.018	0.004	0.284	0.017	0.005	0.477	0.018	0.006	0.737
p-value	.011	.015	.940	.360	.546	.998	.872	.652	.025	.034	.048	.878

*Means within a column with different superscripts are significantly different (p < .05).

NC: Non-injected; PC: Diluent-injected controls; MnS: Manganese sulphate; Mn-Met: Manganese-methionine; NMn: nano manganese; NMn-Met: nano manganese–methionine; E18: embryonic day 18; P1: post-hatch days 1; P7: post-hatch days 7; P14: post-hatch days 14.

Table 5. Effect of injection of different sources of manganese on bone breaking strength (kg) on embryonic day 18 and post-hatch days 1, 7, 14*.

Treatments	E18 (kg)	P1 (kg)	P7 (kg)	P14 (kg)
NC	0.401	0.944	2.872	5.752
PC	0.403	0.913	2.886	5.658
MnS	0.403	0.938	2.877	5.772
Mn-Met	0.404	0.969	2.955	5.801
NMn	0.405	0.954	2.957	5.845
NMn-Met	0.409	0.996	2.941	5.772
SEM	0.006	0.012	0.025	0.042
p-value	.910	.236	.827	.801

*Means within a column with different superscripts are significantly different (p < .05).

NC: Non-injected; PC: Diluent-injected controls; MnS: Manganese sulphate; Mn-Met: Manganese-methionine; NMn: nano manganese; NMn-Met: nano manganese –methionine; E18: embryonic day 18; P1: post-hatch days 1; P7: post-hatch days 7; P14: post-hatch days 14.

Table 6. Effect of injection of different sources of manganese on percentage of Mn in the tibiae ash on embryonic day 18 and post-hatch days 1, 7, 14*.

Treatments	E18 (%)	P1 (%)	P7 (%)	P14 (%)
NC	0.009^c	0.005^c	0.005^c	0.005^c
PC	0.008^c	0.005^c	0.005^c	0.005^c
MnS	0.011^b	0.008^ab	0.005^c	0.008^b
Mn-Met	0.011^b	0.007^b	0.006^bc	0.008^b
NMn	0.012^a	0.009^a	0.010^a	0.010^a
NMn-Met	0.012^a	0.009^a	0.007^b	0.009^ab
SEM	0.0002	0.0002	0.0002	0.0002
p-value	.0001	.0001	.0001	.0001

*Means within a column with different superscripts are significantly different (p < .05).

NC: Non-injected; PC: Diluent-injected controls; MnS: Manganese sulphate; Mn-Met: Manganese-methionine; NMn: nano manganese; NMn-Met: nano manganese –methionine; E18: embryonic day 18; P1: post-hatch days 1; P7: post-hatch days 7; P14: post-hatch days 14.