APSS 2013 Proceedings - The University of Sydney

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Aust. Poult. Sci. Symp. 2013.....24 Broiler premix (d5 – wk 11) and Alltech PN Pre-Harvest Broiler premix modified to contain 2.0 g/kg available P and 7.0 g/kg Ca (wk 11 – 17, Conditioning-PN). At 17 weeks of age, eight birds from each treatment were randomly selected and euthanized by argon asphyxiation followed by cervical dislocation. The small intestine was quickly removed, rinsed to remove contents and a sample from jejunum section 3cm proximal to Meckel's diverticulum was flash frozen in liquid nitrogen and stored at -80°C until further analysis. Total RNA was isolated from the jejunum using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s suggested protocol. Total RNA (0.5 μg) was reverse transcribed into cDNA using the High Capacity cDNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA) according to the manufacturer’s instructions. Real-time PCR was performed in triplicate using Taqman Gene Expression Assays (Applied Biosystems) and the 7500 Real-time PCR system (Applied Biosystems). Ring finger protein 4 (RNF4) was selected as an endogenous control gene to account for any variation in the efficiency of reverse transcription and PCR. The relative quantification (RQ) was expressed as a ratio of the target gene to control gene using the delta-delta Ct ( ΔΔ Ct) method. Data presented are least square means (lsmeans) ±SEM. Real-time PCR results for Normal-PN and Conditioning-PN were compared with ANOVA analysis for a completely randomized experimental design using the Proc GLM procedure of the SAS version 9.1 statistical package (SAS Inst. Inc., Cary, NC, USA). Comparisons of the treatments to the control were made using a Student’s t-test. III. RESULTS AND DISCUSSION Relative expression of zinc transport protein (ZnT) 1 and 7 did not differ between treatments (Figure 1). Expression of ZnT4 was increased in both Conditioning-PN- and Normal-PN-fed birds than control (P < 0.05), but did not differ between birds fed the Conditioning-PN and Normal-PN treatments. Normal-PN fed birds had greater ZnT5 and ZnT6 mRNA levels compared with control (P < 0.05), but levels did not differ from the Conditioning-PN-fed birds. Relative mRNA levels of ferritin, heavy polypeptide 1 (FTH1), metallothionein 3 (MT3) and metallothionein 4 (MT4) did not differ between treatments (Figure 2). The posthatch conditioning supplement had no effect on calbindin 1(CALB1) mRNA levels, however, expression was greater in both Conditioning-PN- and Normal-PN-fed birds than control at 17 weeks of age (P < 0.05). Calbindin 1 levels track directly with changes in Ca absorption with a correlation coefficient of 0.99 (Morrissey and Wasserman, 1971). Therefore birds fed either treatment in this study would likely result in an increase in Ca absorption. Somewhat surprisingly, solute carrier family 34 (sodium phosphate) member 2 (NaPi-IIb) mRNA levels were reduced 1.37-fold (P = 0.03) in Conditioning-PN birds compared with the control. When dietary P levels are reduced, there is an adaptive response in which NaPi-IIb mRNA abundance increases (Giral et al., 2009), therefore it would be expected that NaPi-IIb mRNA abundance would be greatest in both the Conditioning-PN- and Normal-PN-fed birds where available P levels were reduced in the starter and grower phases. This may also indicate that P release by the enzyme inclusion, which among others provides 300 SPU/kg of phytase activity, in the diets containing Alltech PN premix resulted in the down regulation of NaPi-2b to regulate P uptake when more P was available. IV. CONCLUSION Results from this study indicate that, in layers, the expression of some nutrient carrier and transport genes can be influenced by a post-hatch preconditioning period. Research from this lab has shown that diets fed in early life (4 d post-hatch) can have lifelong effects on expression patterns of nutrient carriers and transporters in the intestine regardless of diets fed 204
Aust. Poult. Sci. Symp. 2013.....24 later in life (Brennan et al., 2012). These results indicates that diets formulated using the Alltech PN Grower (fed d5-wk 11) and Pre-Harvest (fed wk11-17) Broiler premixes but not Figure 1 - Relative expression of zinc transport protein (ZnT) in the jejunum of pullets fed control (CON), Cornsoy diet for 4 d followed by diets formulated with the Alltech PN premixes (Normal-PN), and Alltech post-hatch conditioning supplement for 4 d followed by diets formulated with the Alltech PN premixes (Conditioning-PN) using real-time PCR. Values represented are least square means ± SEM; n = 8. *Columns without common superscripts differ at P ≤ 0.05 Figure 2 - Relative expression of nutrient transport genes in the jejunum of pullets fed control (CON), Corn-soy diet for 4 d followed by diets formulated with the Alltech PN premixes (Normal-PN), and Alltech posthatch conditioning supplement for 4 d followed by diets formulated with the Alltech PN premixes (Conditioning-PN) using real-time PCR. Values represented are least square means ± SEM; n = 8. *Columns without common superscripts differ at P ≤ 0.05 the post-hatch conditioning diet, has an effect on nutrient transporter gene expression in the jejunum. This preliminary study helps us better understand nutrient-gene interactions in poultry and how different dietary strategies can influence the expression of genes involved in mineral uptake and binding. 205
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24 th ANNUAL AUSTRALIAN POULTRY SCI
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AUSTRALIAN POULTRY SCIENCE SYMPOSIU
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AUSTRALIAN POULTRY AWARD The Austra
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CONTENTS THE IMPORTANCE OF THE POUL
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SHORT COMMUNICATION SESSION # 1: (c
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SHORT COMMUNICATION SESSTION # 2: (
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HOT TOPIC 2: EMERGING DISEASES AND
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Aust. Poult. Sci. Symp. 2013.....24
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INTERACTIVE EFFECTS OF EXOGENOUS EN
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microbiota to favor an increased ac
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AUTHOR INDEX Name Page(s) Email Add
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Klein-Hessling, H 207 hkleinhesslin
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Van de Wouw, A 247 nnet.vandewouw@w
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APSS 2013 Proceedings - The University of Sydney

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