Mapping the carbon footprint of milk production from cattle: A systematic review.

Recently, many studies have assessed the carbon footprint of bovine milk production. However, due to the complexity of life cycle assessment, most studies have analyzed research farms or "representative" farms, which do not capture farm variability. Furthermore, the lack of consistency in most studies means that we can seldom compare the footprint between different countries. To address this issue, we performed a systematic review of the literature, removing inconsistencies in life cycle assessment papers, namely the functional unit, allocation to milk, and global warming potential. We analyzed papers that accounted for many farms to address the variability of production systems within the countries. We found 21 papers from 19 countries; footprint recalculations were necessary for 16 papers. New Zealand, Uruguay, the United Kingdom, Australia, and the United States had a footprint <1 kg of carbon dioxide equivalents (CO2e) per kilogram of fat- and protein-corrected milk, whereas 5 countries had a footprint >2 kg CO2e·kg-1 fat- and protein-corrected milk. The change in functional unit resulted in a small effect on the final footprint, whereas the global warming potential change was dependent in on the greenhouse gas profile for each country. Countries where milk is produced mainly as a pasture-based system had most of their footprint (>50%) associated with the emission of methane from enteric fermentation, whereas other countries (especially from Europe and North America) had a significant share of emissions from manure management, feed production, and fertilizer use. This different greenhouse gas profile allow decision makers to tailor mitigation options specific for each country. The choice of the allocation method had a strong influence in the final footprint. We suggest that for future studies, authors adhere to the International Dairy Federation guidelines. When this is not possible, we suggest a set of extra information to be reported, allowing recalculations as done in this review.

IGF1 does not overcome sexual dimorphism of body and muscle size in Mstn-/- mice.

Insulin-like growth factor-1 (IGF1) is crucial for regulating post-natal growth and, along with myostatin (MSTN), regulates muscle size. Here, we sought to clarify the roles of these two genes in regulating sexually dimorphic growth of body and muscle mass. In the first study, we established that Igf1 mRNA was increased to a greater extent and Igf1 receptor mRNA increased earlier in male, than in female, gastrocnemius muscles during the rapid phase of growth (from 2 to 6 weeks) were unchanged, thereafter, to 32 weeks of age in WT mice (P < 0.001). In the second study, we sought to determine if supplemental IGF1 could overcome the sexual dimorphism of muscle and body mass, when myostatin is absent. We crossed myostatin null (Mstn-/-) mice with mice over-expressing Igf1 in skeletal muscle (Igf1+) to generate six genotypes; control (Mstn+/+), Mstn+/-, Mstn-/-, Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+ (n = 8 per genotype and sex). In both sexes, body mass at 12 weeks was increased by at least 1.6-fold and muscle mass by at least 3-fold in Mstn-/-:Igf1+ compared with Mstn+/+ mice (P < 0.001). The abundance of AKT was increased in muscles of mice transgenic for Mstn, while phosphorylation of AKTS473 was increased in both male and female mice transgenic for Igf1+. The ratio of phosphorylated to total AKT was 1.9-fold greater in male mice (P < 0.001). Thus, despite increased growth of skeletal muscle and body size when myostatin was absent and IGF1 was in excess, sexual dimorphism persisted, an effect consistent with greater IGF1-induced activation of AKT in skeletal muscles of males.

Eutrophication and climate change impacts of a case study of New Zealand beef to the European market.

OBJECTIVE: Beef production in the Lake Taupo region of New Zealand (NZ) is regulated for nitrogen (N) leaching. The objectives of this study were to 1) evaluate the implications of nitrogen emission limitations on eutrophication and climate change impacts of NZ beef through its life cycle to a European market and uniquely link it to 2) estimation of the reduction in these impacts that can be funded by the consumer's willingness to pay (WTP) a premium for a low environmental-impact product. METHOD: The cradle-to-market Life Cycle Assessment (LCA) of NZ beef on the European market included beef production on farms, meat processing, packaging and transport stages. Various beef production systems in the Lake Taupo region were modelled: farm systems with and without regulated N leaching limits in place (using N fertiliser inputs of 0 and 100 kg N/ha/year respectively) using suckler beef or beef derived from surplus calves from a dairy farm. The FARMAX model was used to model farm productivity and profitability under these various scenarios, whereas the OVERSEER® model was used to model field/farm emissions (N, phosphorus (P)) and the NZ greenhouse gas (GHG) Inventory model was used to estimate total GHG emissions. Eutrophication and climate change impacts of NZ beef to the European market were calculated using recent regionalised LCA indicators. We estimated freshwater and marine eutrophication impacts of European beef using published N emissions to water and air. We estimated the European consumer's WTP for beef with positive environmental attributes based on a meta-regression analysis based on 21 published studies and compared farmer's profit for the farm system scenarios. RESULTS: When using common P-driven eutrophication indicators, the farms using 100 kg fertiliser-N/ha/year appeared to have a lower freshwater eutrophication impact than farms using no N fertiliser, which is in contradiction with the local freshwater policy for N regulations. When the contribution of both N and P were accounted for, the farms using no N fertiliser had the lowest estimated impact. Comparison with published environmental footprint of beef from Europe showed lower climate change and eutrophication impacts for NZ beef, thus showing potential positive environmental attributes for NZ beef. The European consumer's WTP (32% price premium) for such a beef product with low environmental impacts could offset the cost to farmers for implementing the reduction of N emissions. CONCLUSIONS: Bridging the gap between local freshwater policy and LCA indicators starts by considering both P and N emissions and impacts. Combining an environmental LCA with an economic analysis revealed that the consumer willingness to pay could compensate for the environmental cost of protecting the lake that currently only the farmers are bearing.

Water scarcity footprint of dairy milk production in New Zealand - A comparison of methods and spatio-temporal resolution.

Water scarcity footprinting now has a consensual life cycle impact assessment indicator recommended by the UNEP/SETAC Life Cycle Initiative called AWaRe. It was used in this study to calculate the water scarcity footprint of New Zealand (NZ) milk produced in two contrasting regions; "non-irrigated moderate rainfall" (Waikato) and "irrigated low rainfall" (Canterbury). Two different spatial and temporal resolutions for the inventory flows and characterisation factors (CFs) were tested and compared: country and annual vs. regional and monthly resolution. An inventory of all the consumed water flows was carried out from cradle to farm-gate, i.e. from the production of dairy farm inputs to the milk and meat leaving the dairy farm, including all water uses on-farm such as irrigation water, cow drinking water and cleaning water. The results clearly showed the potential overestimation of a water scarcity footprint when using aggregated CFs. Impacts decreased by 74% (Waikato) and 33% (Canterbury) when regional and monthly CFs were used instead of country and annual CFs. The water scarcity footprint calculated at the regional and monthly resolution was 22 Lworld eq/kg FPCM (Fat Protein Corrected Milk) for Waikato milk, and 1118 Lworld eq/kg FPCM for Canterbury milk. The contribution of background processes dominated for milk from non-irrigated pasture, but was negligible for milk from irrigated pasture, where irrigation dominated the impacts. Results were also compared with the previously widely-used Pfister method (Pfister et al., 2009) and showed very similar ranking in terms of contribution analysis. An endpoint indicator was evaluated and showed damages to human health of 7.66 × 10-5 DALY/kg FPCM for Waikato and 2.05 × 10-3 DALY/kg FPCM for Canterbury, but the relevance of this indicator for food production needs reviewing. To conclude, this study highlighted the importance of using high-resolution CFs rather than aggregated CFs.

IGF1 stimulates greater muscle hypertrophy in the absence of myostatin in male mice.

Insulin-like growth factors (IGFs) and myostatin have opposing roles in regulating the growth and size of skeletal muscle, with IGF1 stimulating, and myostatin inhibiting, growth. However, it remains unclear whether these proteins have mutually dependent, or independent, roles. To clarify this issue, we crossed myostatin null (Mstn-/-) mice with mice overexpressing Igf1 in skeletal muscle (Igf1+) to generate six genotypes of male mice; wild type (Mstn+/+ ), Mstn+/-, Mstn-/-, Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+ Overexpression of Igf1 increased the mass of mixed fibre type muscles (e.g. Quadriceps femoris) by 19% over Mstn+/+ , 33% over Mstn+/- and 49% over Mstn-/- (P < 0.001). By contrast, the mass of the gonadal fat pad was correspondingly reduced with the removal of Mstn and addition of Igf1 Myostatin regulated the number, while IGF1 regulated the size of myofibres, and the deletion of Mstn and Igf1+ independently increased the proportion of fast type IIB myosin heavy chain isoforms in T. anterior (up to 10% each, P < 0.001). The abundance of AKT and rpS6 was increased in muscles of Mstn-/-mice, while phosphorylation of AKTS473 was increased in Igf1+mice (Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+). Our results demonstrate that a greater than additive effect is observed on the growth of skeletal muscle and in the reduction of body fat when myostatin is absent and IGF1 is in excess. Finally, we show that myostatin and IGF1 regulate skeletal muscle size, myofibre type and gonadal fat through distinct mechanisms that involve increasing the total abundance and phosphorylation status of AKT and rpS6.

Translational signalling, atrogenic and myogenic gene expression during unloading and reloading of skeletal muscle in myostatin-deficient mice.

Skeletal muscles of myostatin null (Mstn(-/-)) mice are more susceptible to atrophy during hind limb suspension (HS) than are muscles of wild-type mice. Here we sought to elucidate the mechanism for this susceptibility and to determine if Mstn(-/-) mice can regain muscle mass after HS. Male Mstn(-/-) and wild-type mice were subjected to 0, 2 or 7 days of HS or 7 days of HS followed by 1, 3 or 7 days of reloading (n = 6 per group). Mstn(-/-) mice lost more mass from muscles expressing the fast type IIb myofibres during HS and muscle mass was recovered in both genotypes after reloading for 7 days. Concentrations of MAFbx and MuRF1 mRNA, crucial ligases regulating the ubiquitin-proteasome system, but not MUSA1, a BMP-regulated ubiquitin ligase, were increased more in muscles of Mstn(-/-) mice, compared with wild-type mice, during HS and concentrations decreased in both genotypes during reloading. Similarly, concentrations of LC3b, Gabarapl1 and Atg4b, key effectors of the autophagy-lysosomal system, were increased further in muscles of Mstn(-/-) mice, compared with wild-type mice, during HS and decreased in both genotypes during reloading. There was a greater abundance of 4E-BP1 and more bound to eIF4E in muscles of Mstn(-/-) compared with wild-type mice (P<0.001). The ratio of phosphorylated to total eIF2α increased during HS and decreased during reloading, while the opposite pattern was observed for rpS6. Concentrations of myogenic regulatory factors (MyoD, Myf5 and myogenin) mRNA were increased during HS in muscles of Mstn(-/-) mice compared with controls (P<0.001). We attribute the susceptibility of skeletal muscles of Mstn(-/-) mice to atrophy during HS to an up- and downregulation, respectively, of the mechanisms regulating atrophy of myofibres and translation of mRNA. These processes are reversed during reloading to aid a faster rate of recovery of muscle mass in Mstn(-/-) mice.

Identification and expression of a novel transcript of the growth and differentiation factor-11 gene.

The growth and differentiation factor-11 (GDF-11) gene is thought to code for a single protein that plays a crucial role in regulating the development of multiple tissues. In this study, we aimed to investigate if the GDF-11 gene has another transcript and, if so, to characterise this transcript and determine its tissue-specific and developmental expression. We have identified a novel transcript of GDF-11 in mouse muscle, which contains the 3' region of intron 1, exon 2, exon 3 and 3'UTR, and has two transcription initiation sites and a single termination site. We named the novel transcript GDF-11ΔEx1 because it does not contain exon 1 of canonical GDF-11. The GDF-11ΔEx1 transcript was expressed in the skeletal muscles, heart, brain and kidney, but was undetectable in the liver and gut. The concentration of the GDF-11ΔEx1 transcript was increased in gastrocnemius muscles from three to 6 weeks of age, a period of accelerated muscle growth, steadily declined thereafter and was higher in male than female mice (P < 0.001 for age and sex). GDF-11ΔEx1 cDNA was predicted to code for a putative N-terminal-truncated propeptide and the canonical ligand for GDF-11. However, propeptide-specific antibodies could not identify proteins of the expected size in skeletal muscle. Interestingly, in silico analysis of the GDF-11ΔEx1 RNA predicted a secondary structure with the potential to coordinate multiple protein interactions as a molecular scaffold. Therefore, we postulate that GDF-11ΔEx1 may act as a long non-coding RNA to regulate the transcription of canonical GDF-11 and/or other genes in skeletal muscle and other tissues.

Discovery of a mammalian splice variant of myostatin that stimulates myogenesis.

Myostatin plays a fundamental role in regulating the size of skeletal muscles. To date, only a single myostatin gene and no splice variants have been identified in mammals. Here we describe the splicing of a cryptic intron that removes the coding sequence for the receptor binding moiety of sheep myostatin. The deduced polypeptide sequence of the myostatin splice variant (MSV) contains a 256 amino acid N-terminal domain, which is common to myostatin, and a unique C-terminus of 65 amino acids. Western immunoblotting demonstrated that MSV mRNA is translated into protein, which is present in skeletal muscles. To determine the biological role of MSV, we developed an MSV over-expressing C2C12 myoblast line and showed that it proliferated faster than that of the control line in association with an increased abundance of the CDK2/Cyclin E complex in the nucleus. Recombinant protein made for the novel C-terminus of MSV also stimulated myoblast proliferation and bound to myostatin with high affinity as determined by surface plasmon resonance assay. Therefore, we postulated that MSV functions as a binding protein and antagonist of myostatin. Consistent with our postulate, myostatin protein was co-immunoprecipitated from skeletal muscle extracts with an MSV-specific antibody. MSV over-expression in C2C12 myoblasts blocked myostatin-induced Smad2/3-dependent signaling, thereby confirming that MSV antagonizes the canonical myostatin pathway. Furthermore, MSV over-expression increased the abundance of MyoD, Myogenin and MRF4 proteins (P<0.05), which indicates that MSV stimulates myogenesis through the induction of myogenic regulatory factors. To help elucidate a possible role in vivo, we observed that MSV protein was more abundant during early post-natal muscle development, while myostatin remained unchanged, which suggests that MSV may promote the growth of skeletal muscles. We conclude that MSV represents a unique example of intra-genic regulation in which a splice variant directly antagonizes the biological activity of the canonical gene product.

The decrease in mature myostatin protein in male skeletal muscle is developmentally regulated by growth hormone.

Myostatin inhibits myogenesis and there is reduced abundance of the mature protein in skeletal muscles of adult male compared with female mice. This reduction probably occurs after translation, which suggests that it is a regulated mechanism to reduce the availability of myostatin in males. Reduced myostatin may, thereby, contribute to the development of sexually dimorphic growth of skeletal muscle. Our first objective was to determine if the decrease in mature myostatin protein occurs before the linear growth phase to aid growth, or afterwards to maintain the mass of adult muscle. Mice were killed from 2 to 32 weeks and the gastrocnemius muscle was excised. Myostatin mRNA increased from 2 to 32 weeks and was higher in males than females (P < 0.001). In contrast, mature protein decreased in males after 6 weeks (P < 0.001). Our second objective was to determine if growth hormone (GH) induces the decrease in mature myostatin protein. GH increased myostatin mRNA and decreased the abundance of mature protein in hypophysectomised mice (P < 0.05). Our final objective was to determine if the decrease in mature protein occurs in skeletal muscles of male Stat5b(-/-) mice (Stat5b mediates the actions of GH). As expected, mature myostatin protein was not reduced in Stat5b(-/-) males compared with females. However, myostatin mRNA remained higher in males than females irrespective of genotype. These data suggest that: (1) the decrease in mature myostatin protein is developmentally regulated, (2) GH acting via Stat5b regulates the abundance of mature myostatin and (3) GH acts via a non-Stat5b pathway to regulate myostatin mRNA.

Effect of botulinum toxin A-induced paralysis and exercise training on mechanosensing and signalling gene expression in juvenile rat gastrocnemius muscle.

Intramuscular injections of the paralytic botulinum neurotoxin A (Btx) and physical exercise are used in the treatment of chronic spasticity in children with cerebral palsy. We tested whether Btx-induced paralysis and/or exercise training would have differential effects on the expression of mechanosensing and signalling genes implicated in the adaptive remodelling of skeletal muscle. Juvenile (29-day-old) male rats were injected with Btx or saline (NoBtx) into the right gastrocnemius and housed in standard cages (NoEx) or with running wheels (Ex), for 3 weeks (n = 6 per group). The mRNA expression of nine sarcomere-associated genes in the medial gastrocnemius was then determined by quantitative reverse transcriptase-polymerase chain reaction. The Btx-injected muscles weighed 50% less than NoBtx muscles, but Ex had no effect on the wet mass of Btx or NoBtx muscles. Atrogenic MuRF1, sarcomeric Titin and myogenic MyoD were upregulated (2-fold) with the elimination of contractile activity in Btx muscle. Expression of CARP, Ankrd2 and MLP was increased with mechanical stimuli associated with Btx (5- to 10-fold) or Ex (2- to 4-fold). Expression of CARP and Ankrd2 increased synergistically in Btx-Ex muscle (> or = 20-fold), indicating that these genes may be sensitive to passive stretch of the sarcomeric I-band region of titin to which their proteins bind. Tcap, Myopalladin and Atrogin1 were not, or were no longer responsive to the altered mechanical stimuli after 3 weeks of Btx or Ex. The expression of Ankrd2, CARP and MLP may thus be enhanced by passive stretch within the Btx-paralysed and/or exercising gastrocnemius and contribute to adaptations, other than muscle mass, in juvenile rats.