Roy G. Meidinger

Pigs expressing salivary phytase produce low-phosphorus manure

To address the problem of manure-based environmental pollution in the pork industry, we have developed the phytase transgenic pig. The saliva of these pigs contains the enzyme phytase, which allows the pigs to digest the phosphorus in phytate, the most abundant source of phosphorus in the pig diet. Without this enzyme, phytate phosphorus passes undigested into manure to become the single most important manure pollutant of pork production. We show here that salivary phytase provides essentially complete digestion of dietary phytate phosphorus, relieves the requirement for inorganic phosphate supplements, and reduces fecal phosphorus output by up to 75%. These pigs offer a unique biological approach to the management of phosphorus nutrition and environmental pollution in the pork industry.

Application of iTRAQ to catalogue the skeletal muscle proteome in pigs and assessment of effects of gender and diet dephytinization.

In this study iTRAQ was used to produce a highly confident catalogue of 542 proteins identified in porcine muscle (false positive<5%). To our knowledge this is the largest reported set of skeletal muscle proteins in livestock. Comparison with human muscle proteome demonstrated a low level of false positives with 83% of the proteins common to both proteomes. In addition, for the first time we assess variations in the muscle proteome caused by sexually dimorphic gene expression and diet dephytinization. Preliminary analysis identified 19 skeletal muscle proteins differentially expressed between male and female pigs (≥1.2‐fold, p<0.05), but only one of them, GDP‐dissociation inhibitor 1, was significant (p<0.05) after false discovery rate correction. Diet dephytinization affected expression of 20 proteins (p<0.05). This study would contribute to an evaluation of the suitability of the pig as a model to study human gender‐related differences in gene expression. Transgenic pigs used in this study might also serve as a useful model to understand changes in human physiology resulting from diet dephytinization.

A manganese superoxide dismutase-encoding cDNA from Drosophila melanogaster

A cDNA encoding manganese superoxide dismutase (MnSOD) has been cloned and sequenced from Drosophila melanogaster. This 805-bp complete cDNA carries a single open reading frame which specifies a deduced 217-amino-acid (aa) protein with an average identity of 60.8% with deduced MnSOD aa sequences from Caenorabditis elegans and humans.

Digestive utilization of phosphorus from plant-based diets in the Cassie line of transgenic Yorkshire pigs that secrete phytase in the saliva

A line of transgenic Yorkshire pigs referred to as the Cassie (CA) line was generated, which possessed a stable, low copy number phytase transgene insertion that enabled phytase secretion in the saliva. This study was conducted to assess growth and efficacy for improving P, Ca, and other macromineral utilization in the CA pigs receiving diets typical of those used for commercial swine production. In Exp. 1, 12 CA boars and 12 CA gilts fed diets without supplemental P gained weight and exhibited feed efficiency similar to conventional age-matched 12 Yorkshire boars and 12 Yorkshire gilts raised on similar diets with supplemental P. Serum concentrations of P and Ca were similar for CA and Yorkshire pigs during the growing and finishing phases, indicating that the CA pigs were not P limited. In Exp. 2, 6 CA (13.1 kg BW) and 6 Yorkshire barrows (8.8 kg BW) were fed 3 diets (control; low in Ca and P; and low in Ca, P, and CP) over 3 phases. The CA barrows fed the diet without supplemental P retained 25 to 40% (P < 0.001), 77 to 91% (P < 0.001), and 27 to 56% (P < 0.001) more P during the weaning, growing, and finishing phases, respectively, than conventional Yorkshire barrows fed similar diets without supplemental P. In Exp. 3, CA and Yorkshire barrows of similar ages weighing 66.2 ± 1.7 kg (n = 10) and 50.0 ± 1.0 kg (n = 10), respectively, were used. The P retention of CA finisher barrows fed a diet without supplemental P was 34% greater (P < 0.001) than conventional Yorkshire barrows fed the same diet with 750 units of exogenous phytase/kg diet. Urinary Ca to P ratio in the CA pigs was 0.27, whereas that for the Yorkshire barrows was 30, thereby, indicating that the Yorkshire barrows suffered a P deficiency. Furthermore, digestive utilization of major electrolyte macrominerals, K and Na, was improved (P < 0.05) by 18 and 16%, respectively, in the CA finisher pigs compared with the conventional Yorkshire finisher pigs fed phytase; however, only K exhibited enhanced retention. In conclusion, the CA line pigs secrete sufficient phytase from the salivary glands to enable efficient digestion of plant P, Ca, and major electrolyte macrominerals.

Genetic Opportunities tyo Enhance Sustainability of Pork Production in Developing Countries: A Model for Food Animals

Currently there is a shortage of food and potable water in many developing countries. Superimposed upon this critical situation, because of the increasing urban wealth in these countries, there is a strong trend of increased consumption of meat, and pork in particular. The consequence of this trend will be increased agricultural pollution, resulting not only from greater use of chemical fertilizer, but also from manure spread on land as fertilizer that may enter freshwater and marine ecosystems causing extensive eutrophication and decreased water quality. The application of transgenic technologies to improve the digestive efficiency and survival of food animals, and simultaneously decreasing their environmental impact is seen as an opportunity to enhance sustainability of animal agriculture without continued capital inputs. Transgenes expressed in pigs that have potential include, for example, genes coding for phytase, lactalbumin and lactoferrin. At the University of Guelph, Escherichia coli phytase has been expressed in the salivary glands of the pig. Selected lines of these pigs utilize plant phytate phosphorus efficiently as a source of phosphorus and excrete faecal material with more than a 60 percent reduction in phosphorus content. Because of their capacity to utilize plant phytate phosphorus and to produce less polluting manure they have a valuable trait that will contribute to enhanced sustainability of pork production in developing countries, where there is less access to either high quality phosphate supplement or phytase enzyme to include in the diet. Issues that require continued consideration as a prelude to the introduction of transgenic animals into developing countries include food and environmental safety, and consumer acceptance of meat products from genetically modified animals.

Comparative carcass and tissue nutrient composition of transgenic Yorkshire pigs expressing phytase in the saliva and conventional Yorkshire pigs

A transgenic line of Yorkshire (YK) pigs named the Cassie (CA) line was produced with a low copy number phytase transgene inserted in the genome. The transgenic line efficiently digests P, Ca, and other major minerals of plant dietary origin. The objectives of this study were to 1) compare carcass and tissue nutrient composition and meat quality traits for third generation hemizygous CA line market BW finisher pigs (n = 24) with age-matched conventional YK finisher pigs (n = 24) and 2) examine effects of outbreeding with high-index conventional YK boars on modifying carcass leanness from the third to sixth generations in CA line finisher boars (n = 73) and gilts (n = 103). Cassie boars (n = 12) and CA gilts (n = 12) were fed diets without supplemental P and comparable numbers of age-matched YK boars and gilts fed diets containing supplement P were raised throughout the finisher phase. The pigs were slaughtered and then fabricated into commercial pork primals before meat composition and quality evaluation. Proximate and major micronutrient composition was determined on tissues including fat, kidney, lean, liver, and skin. The main difference observed was greater (P = 0.033) crude fat content in CA boar carcasses and increased (P < 0.04) leaf lard in both CA boars and gilts but no differences were observed (P = 0.895 and P = 0.223, respectively) in carcass backfat thickness as compared with YK pigs. There were no substantive differences in tissue composition, except for CA boar kidneys. Numerous changes in the mineral, fatty acid, and indispensable AA composition for CA boar kidneys were not apparent in CA gilts. These changes may point to adaptive physiological changes in the boar kidney necessary for homeostatic regulation of mineral retention related to phytase action rather than to insertion of the transgene. However, from a meat composition perspective, transgenic expression of phytase in the CA line of YK pigs had little overall effect on meat composition. Outbreeding of high-index CA gilts with high-index commercial YK boars linearly reduced (P = 0.002) back fat thickness with a corresponding linear increase (P = 0.001) in lean yield in finisher CA gilts, although no change in these parameters was observed in CA finisher boars. The increase in lean yield in CA gilts by selective breeding without affecting the level of salivary phytase activity documents the value of conventional genetic selection in conjunction with genetic modification.