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Dairy cow subfertility is a worldwide issue arising from multiple factors. It manifests in >30% early pregnancy losses in seasonal pasture-grazed herds, especially when cows are inseminated in the early post-partum period. Most losses occur before implantation, when embryo growth depends on factors present in maternal tract fluids. Here we examined the proteomic composition of early and mid-postpartum uterine luminal fluid (ULF) in crossbred lactating dairy cows to identify molecular determinants of fertility. We also explored changes in ULF from first to third estrus cycles postpartum in individual cows, linking those changes with divergent embryo development. For this, we flushed uteri of 87 cows at Day 7 of pregnancy at first and third estrus postpartum, recovering, and grading their embryos. Out of 1563 proteins detected, 472 had not been previously reported in this fluid, and 408 were predicted to be actively secreted by bioinformatic analysis. The abundance of 18 proteins with roles in immune regulation and metabolic function (e.g. cystatin B, pyruvate kinase M2) was associated with contrasting embryo quality. Matched-paired pathway analysis indicated that, from first to third estrus postpartum, upregulation of metabolic (e.g. creatine and carbohydrate) and immune (e.g. complement regulation, antiviral defense) processes were related to poorer quality embryos in the third estrus cycle postpartum. Conversely, upregulated signal transduction and protein trafficking appeared related to improved embryo quality in third estrus. These results advance the characterization of the molecular environment of bovine ULF and may aid understanding fertility issues in other mammals, including humans.

Egg and egg products are a rich source of highly bioavailable animal proteins. Several processing technologies can affect the structural and functional properties of these proteins differently and can influence their fate inside the gastrointestinal tract. The present review examines some of the processing technologies for improving egg protein digestibility and discusses how different processing conditions affect the digestibility of egg proteins under gastrointestinal digestion environments. To provide up-to-date information, most of the studies included in this review have been published in the last 5 years on different aspects of egg protein digestibility. Digestibility of egg proteins can be improved by employing some processing technologies that are able to improve the susceptibility of egg proteins to gastrointestinal proteases. Processing technologies, such as pulsed electric field, high-pressure, and ultrasound, can induce conformational and microstructural changes that lead to unfolding of the polypeptides and expose active sites for further interactions. These changes can enhance the accessibility of digestive proteases to cleavage sites. Some of these technologies may inactivate some egg proteins that are enzyme inhibitors, such as trypsin inhibitors. The underlying mechanisms of how different technologies mediate the egg protein digestibility have been discussed in detail. The proteolysis patterns and digestibility of the processed egg proteins are not always predictable and depends on the processing conditions. Empirical input is required to tailor the optimization of processing conditions for favorable effects on protein digestibility.

Thermal processing is an inevitable part of the processing and preparation of meat and meat products for human consumption. However, thermal processing techniques, both commercial and domestic, induce modifications in muscle proteins which can have implications for their digestibility. The nutritive value of muscle proteins is closely related to their digestibility in the gastrointestinal tract and is determined by the end products that it presents in the assimilable form (amino acids and small peptides) for the absorption. The present review examines how different thermal processing techniques, such as sous-vide, microwave, stewing, roasting, boiling, frying, grilling, and steam cooking, affect the digestibility of muscle proteins in the gastrointestinal tract. By altering the functional and structural properties of muscle proteins, thermal processing has the potential to influence the digestibility negatively or positively, depending on the processing conditions. Thermal processes such as sous-vide can induce favourable changes, such as partial unfolding or exposure of cleavage sites, in muscle proteins and improve their digestibility whereas processes such as stewing and roasting can induce unfavourable changes, such as protein aggregation, severe oxidation, cross linking or increased disulfide (S-S) content and decrease the susceptibility of proteins during gastrointestinal digestion. The review examines how the underlying mechanisms of different processing conditions can be translated into higher or lower protein digestibility in detail. This review expands the current understanding of muscle protein digestion and generates knowledge that will be indispensable for optimizing the digestibility of thermally processed muscle foods for maximum nutritional benefits and optimal meal planning.

Hexaploid Caucasian clover was grown in soil sampled at three New Zealand South Island high country sites to which specific rhizobium inoculum had been added with sowing of the legume in 1975, 1992...

Effect of pulsed electric field (PEF) on in-vitro simulated gastrointestinal protein digestion of cold-boned beef Semimembranosus was elucidated. PEF treated [T1, 5.0 kV (90 Hz); T2, 10 kV (20 Hz)] samples along with control were subjected to in-vitro simulated gastrointestinal protein digestion. Samples collected at 0, 30 and 60 min of gastric digestion and 120 and 180 min of intestinal digestion were analysed for protein digestibility (%), soluble protein (%), protein profile (SDS-PAGE), free amino acid analysis and mineral profile. Significantly higher (P 0.05) of PEF was observed on the release of various minerals from the muscle during the digestion. By modifying the protein profile of the meat digests, PEF processing had a positive influence on in-vitro digestion kinetics that led to a faster and greater digestion of proteins during simulated gastrointestinal digestion.

Although the bioactivities of bovine lactoferrin have been extensively investigated, little is known about deer milk lactoferrin bioactivity and its amino acid sequence. This research investigated the amino acid sequence of deer lactoferrin and the antimicrobial activities of two lactoferrin-encrypted peptides, lactoferricin (Lfcin) and lactoferrampin (Lfampin). Deer lactoferrin was found to have a molecular weight of 77.1 kDa and an isoelectric point of 7.99, which are similar to that of bovine lactoferrin, 78 kDa and pI 7.9. Deer lactoferrin contains 707 amino acids, one amino acid less than bovine lactoferrin, and has 92% homology with bovine lactoferrin. Deer lactoferricin exhibited strong antimicrobial activity against E. coli American Type Culture Collection (ATCC) 25922 and L. acidophilus ATCC 4356. The antimicrobial activities of deer and bovine Lfcin and Lfampin were compared. Based on MIC, deer Lfcin was found to be a more effective inhibitor of L. acidophilus ATCC 4356 than bovine Lfcin, but bovine Lfcin and Lfampin were more effective against E. coli ATCC 25922 than deer Lfcin and Lfampin. The deer Lfcin sequence differed at seven amino acids from bovine Lfcin and this decreased the net positive charge and increased the hydrophobicity. Deer Lfampin contained two differences in amino acid sequence compared to bovine Lfampin which decreased the net positive charge. These amino acid sequence differences likely account for differences in antibacterial activity. Positive charge and hydrophobic residues provide the amphipathic character of these helical peptides, and are considered important for binding of antimicrobial peptides. In silico modelling of deer Lfcin indicated an identical α-helical structure compared to bovine Lfcin.

Meat proteolytic systems play a crucial role in meat tenderisation. Understanding the effects of processing technologies and post-mortem storage conditions on these systems is important due to their crucial role in determining the quality characteristics of meat and meat products. It has recently been proposed that tenderisation occurs due to the synergistic action of numerous endogenous proteolytic systems. There is strong evidence suggesting the importance of μ-calpain during the initial post-mortem aging phase, while m-calpain may have a role during long-term aging. The caspase proteolytic system is also a candidate for cell degradation in the initial stages of conversion of muscle to meat. The role of cathepsins, which are found in the lysosomes, in post-mortem aging is controversial. Lysosomes need to be ruptured, through aging, or other forms of processing to release cathepsins into the cytosol for participation in proteolysis. A combination of optimum storage conditions along with suitable processing may accelerate protease activity within meat, which can potentially lead to improved meat tenderness. Processing technologies such as high pressure, ultrasound, and shockwave processing have been reported to disrupt muscle structure, which can facilitate proteolysis and potentially enhance the aging process. This paper reviews the recent literature on the impacts of processing technologies along with post-mortem storage conditions on the activities of endogenous proteases in meat. The information provided in the review may be helpful in selecting optimum post-mortem meat storage and processing conditions to achieve improved muscle tenderness within shorter aging and cooking times.

Meat tenderness is an important quality trait critical to consumer acceptance, and determines satisfaction, repeat purchase and willingness-to-pay premium prices. Recent advances in tenderness research from a variety of perspectives are presented. Our understanding of molecular factors influencing tenderization are discussed in relation to glycolysis, calcium release, protease activation, apoptosis and heat shock proteins, the use of proteomic analysis for monitoring changes, proteomic biomarkers and oxidative/nitrosative stress. Each of these structural, metabolic and molecular determinants of meat tenderness are then discussed in greater detail in relation to animal variation, postmortem influences, and changes during cooking, with a focus on recent advances. Innovations in postmortem technologies and enzymes for meat tenderization are discussed including their potential commercial application. Continued success of the meat industry relies on ongoing advances in our understanding, and in industry innovation. The recent advances in fundamental and applied research on meat tenderness in relation to the various sectors of the supply chain will enable such innovation.

The effects of egg white protein and soy protein isolate addition on the nutritional and digestibility of gluten-free pasta based on banana flour were studied. The level of protein additions (soy protein or egg white protein) were 0, 5, 10 and 15% of banana flour (w/w). Pasta made from 100% durum wheat semolina was used as a control. Soy protein isolate inclusion into banana pasta increased total phenolic content (TPC) and antioxidant capacities, while egg white protein decreased the TPC and antioxidant capacities with the increasing level of addition. Starch digestibility was affected by the type of protein addition. Egg white protein lowered starch digestibility compared to soy protein isolate. Protein inclusion in banana pasta also altered protein digestibility, amino acid profiles and protein digestibility-corrected amino acid score (PDCAAS). Soy protein isolate increased protein digestibility of gluten-free pasta compared to egg white protein. Protein enrichment gave better amino acid profiles of banana pasta compared to semolina pasta with egg white protein and performed a better PDCAAS compared to soy protein isolate. These results showed that soy protein isolate and egg white protein addition enhanced nutritional qualities and digestibility properties of gluten-free banana pasta.

Cow subfertility is a multi-factorial problem in many countries which is only starting to be unravelled. Molecular biology can provide a substantial source of insight into its causes and potential solutions, particularly through large scale, untargeted omics approaches. In this systematic review, we set out to compile, assess and integrate the latest proteomic and metabolomic research on cow reproduction, specifically that on the female reproductive tract and early embryo. We herein report a general improvement in technical standards throughout the temporal span examined; however, significant methodological limitations are also identified. We propose easily actionable avenues for ameliorating these shortcomings and enhancing the reach of this field. Text mining and pathway analysis corroborate the relevance of proteins and metabolites related to the triad oxidative stress-inflammation-disease on reproductive function. We envisage a breakthrough in cattle reproductive molecular research within the next few years as in vivo sample techniques are improved, omics analysis equipment becomes more affordable and widespread, and software tools for single- and multi-omics data processing are further developed. Additional investigation of the impact of local oxidative stress and inflammation on fertility, both at the local and systemic levels, is key towards realising the full potential of this field.

Aging is a popular method used by meat industry for improving the sensory attributes of meat. Despite the advent of many novel technologies, aging has not lost its charm and is still widely used commercially as a post-mortem intervention for tenderization. Aging improves the tenderness of meat through disruption of the muscle structure by intracellular proteolytic systems. Muscles undergo various molecular changes that cause proteolysis of key myofibrillar and cytoskeletal proteins, disrupting the overall integrity of muscle cells. Although several endogenous proteolytic systems are capable of post-mortem proteolysis, a great body of scientific evidence supports a major role for the calpain system. Calpains are intracellular calcium-dependent cysteine proteases found in most eukaryotes. At least three calpains (μ- and m-calpains and calpain 3) and calpastatin, their specific endogenous inhibitor, are found in muscle. They are known to be involved in the proteolysis of functionally relevant structural proteins such as the myofibrillar proteins and cytoskeletal anchorage complexes. These ubiquitous proteases are also present in mitochondria and play important roles in a variety of pathophysiological conditions including apoptotic and necrotic cell death phenomena. This review discusses the role and contribution of the calpain system and the factors that influence calpain activity during aging. Keywords: Aging, Tenderness, Calpains, Proteolysis, Calpain activity, Factors

The tenderization process, which can be influenced by both pre- and post-slaughter interventions, begins immediately after an animal's death and is followed with the disruption of the muscle structure by endogenous proteolytic systems. The post-slaughter technological interventions like electrical stimulation, suspension methods, blade tenderization, tumbling, use of exogenous enzymes, and traditional aging are some of the methods currently employed by the meat industry for improving tenderness. Over the time, technological advancement resulted in development of several novel methods, for maximizing the tenderness, which are being projected as quick, economical, nonthermal, green, and energy-efficient technologies. Comparison of these advanced technological methods with the current applied industrial methods is necessary to understand the feasibility and benefits of the novel technology. This review discusses the benefits and advantages of different emerging tenderization techniques such as hydrodynamic-pressure processing, high-pressure processing, pulsed electric field, ultrasound, SmartStretch™ , Pi-Vac Elasto-Pack® system, and some of the current applied methods used in the meat industry.

Forty-four rhizobial isolates from New Zealand endemic Sophora species growing in natural conditions were characterised and the effectiveness of selected isolates with regard to growth and N2 fixation of different Sophora species was tested. Sequences for the housekeeping genes (concatenated recA, gln11 and rpoB) of the isolates were diverse but sequences for their symbiosis genes (nifH and nodC) were similar. Generally, isolates from the same field site showed similar housekeeping gene sequences. All isolates were Mesorhizobium. Twenty-six of the isolates aligned with Mesorhizobium species previously isolated from New Zealand native Sophora on the basis of their housekeeping gene sequences, but 18 isolates separated into two groups and a pair of isolates clearly separated from all Mesorhizobium type strains. DNA–DNA hybridisations indicated that isolates in the two groups could belong to two novel Mesorhizobium species. Rhizobial isolates with different housekeeping gene sequences (but similar symbiosis gene sequences) gave similar increases in dry weight and total N content of four Sophora species under glasshouse conditions.

Defined as meat cultured in a laboratory within a bioreactor under controlled artificial conditions, in vitro meat is a relatively recent area that has opened a whole universe of possibilities and opportunities for the meat sector. With improved chemical and microbial safety and varied options, in vitro meat has been proposed as a green, healthy, environmentally friendly, and nutritionally better product that is free from animal suffering and death. Cell culture and tissue culture are the most probable technologies for the development of this futuristic muscle product. However, there are many challenges in the production of a suitable product at an industrial scale under a sustainable production system and a great body of research is required to fill the gaps in our knowledge. Many materials used in the product development are novel and untested within the food industry and demand urgent regulatory and safety assessment systems capable of managing any risks associated with the development of cultured meat. The future of this product will depend on the actions of governments and regulatory agencies. This article highlights emerging biotechnological options for the development of cultured meat and suggests ways to integrate these emerging technologies into meat research. It considers the problems and possibilities of developing cultured meat, opportunities, ethical issues as well as emerging safety and regulatory issues in this area.

Homeoviscous adaptation (HVA) is a key cellular response by which fish protect their membranes against thermal stress. We investigated evolutionary HVA (long time scale) in Antarctic and non-Antarctic fish. Membrane lipid composition was determined for four Perciformes fish: two closely related Antarctic notothenioid species (Trematomus bernacchii and Pagothenia borchgrevinki); a diversified related notothenioid Antarctic icefish (Chionodraco hamatus); and a New Zealand species (Notolabrus celidotus). The membrane lipid compositions were consistent across the three Antarctic species and these were significantly different from that of the New Zealand species. Furthermore, acclimatory HVA (short time periods with seasonal changes) was investigated to determine whether stenothermal Antarctic fish, which evolved in the cold, stable environment of the Southern Ocean, have lost the acclimatory capacity to modulate their membrane saturation states, making them vulnerable to anthropogenic global warming. We compared liver membrane lipid composition in two closely related Antarctic fish species acclimated at 0 °C (control temperature), 4 °C for a period of 14 days in Trematomus bernacchii and 28 days for Pagothenia borchgrevinki, and 6 °C for 7 days in both species. Thermal acclimation at 4 °C did not result in changed membrane saturation states in either Antarctic species. Despite this, membrane functions were not compromised, as indicated by declining serum osmolality, implying positive compensation by enhanced hypo-osmoregulation. Increasing the temperature to 6 °C did not change the membrane lipids of P. borchgrevinki. However, in T. bernacchii, thermal acclimation at 6 °C resulted in an increase of membrane saturated fatty acids and a decline in unsaturated fatty acids. This is the first study to show a homeoviscous response to higher temperatures in an Antarctic fish, although for only one of the two species examined.

The cuticle plays an important role in determining characteristic traits of wool fibres, but little is known about the identity and abundance of its proteinaceous constituents and whether there are unique differences in cuticle composition of wools from different breeds of sheep. In this study, protein profiles of enriched endocuticle fractions from fine (Merino), strong (Romney) wool and a sheep that sheds its wool (Wiltshire) were determined by LC-MS/MS and LC-MALDI, quantitative differences being determined by iTRAQ-labelling. Although keratin composition of the cuticle was similar between breeds, with the exception of K82, there were clear differences in the number of identified keratin-associated proteins. Overall, both qualitative and quantitative differences in protein composition of the cuticle from different sheep breeds were found, which may contribute to the unique characteristics of these wools.

This Data article provides Supplementary data related to the research article titled “In-depth characterisation of the lamb meat proteome from longissimus lumborum ” by Yu et al. [1] . This research article reports the proteome catalogue of the 48 h post-mortem lamb longissimus lumborum. A list of 388 ovine-specific proteins were identified and characterised after separating the samples into sarcoplasmic, myofibrillar and insoluble fractions, followed by an in-depth shotgun proteomic evaluation and bioinformatic analysis. The detailed list of identified proteins, the annotated MS/MS spectra corresponding to the proteins identified by a single peptide-spectrum match, the raw Gene Ontology annotation data and other miscellaneous files, as will be described below, were contained in this Data article. We hope the data presented here will contribute to the current knowledge of the global protein composition of lamb skeletal muscle/meat.

Thaumatin-like proteins (TLPs) and chitinases are the two main groups of pathogenesis-related (PR) proteins found in wine that cause protein haze formation. Previous studies have found that phenolics are also involved in protein haze formation. In this study, Sauvignon Blanc grapes were harvested and processed in two vintages (2011 and 2012) by three different treatments: (1) hand harvesting with whole bunch press (H-WB); (2) hand harvesting with destem/crush and 3 h skin contact (H-DC-3); and (3) machine harvesting with destem/crush and 3 h skin contact (M-DC-3). The juices were collected at three pressure levels (0.4 MPa, 0.8 MPa and 1.6 MPa), some juices were fermented in 750 mL of wine bottles to determine the bentonite requirement for the resulting wines. Results showed juices of M-DC-3 had significantly lower concentration of proteins, including PR proteins, compared to those of H-DC-3, likely due to the greater juice yield of M-DC-3 and interactions between proteins and phenolics. Juices from the 0.8–1.6 MPa pressure and resultant wines had the highest concentration of phenolics but the lowest concentration of TLPs. This supported the view that TLPs are released at low pressure as they are mainly present in grape pulp but additional extraction of phenolics largely present in skin occurs at higher pressing pressure. Wine protein stability tests showed a positive linear correlation between bentonite requirement and the concentration of chitinases, indicating the possibility of predicting bentonite requirement by quantification of chitinases. This study contributes to an improved understanding of extraction of haze-forming PR proteins and phenolics that can influence bentonite requirement for protein stabilization.

There is renewed interest in fodder beet (Beta vulgaris L.) production in the South Island of New Zealand. However, recommended agronomic practices for maximising productivity and quality are limited. Two experiments investigating the effects of different rates (kg/ha) of potassium (K; 150 or 300), sodium (Na; 100 or 200) and chloride (Cl; 240) fertiliser application on fodder beet dry matter (DM) production, nutrient concentration and uptake were carried out at Southbridge (2011) and Bankside (2012), Canterbury, New Zealand. Final DM yield was unaffected by the treatments: averaging 32 t DM/ha at Southbridge and 21 t DM/ha at Bankside. However, there was a significant interaction between K and both Cl and Na, on DM yield at Southbridge. Specifically, yield increased to 38 t DM/ha with Na and decreased to 30 t DM/ha with Cl, in the presence of K. Application of K, Na, and Cl fertiliser increased the respective tissue mineral concentration and respective total uptake for both the bulb and shoot components. Potassium tended to depress crude protein, neutral detergent fibre and ash content of the bulbs and increase the metabolisable energy and soluble sugar. The combination of low fibre and high soluble sugar concentrations puts animals at risk of rumen acidosis. These results suggest that fodder beet crops subjected to high rate of soil and/ or fertiliser K should be supplemented with feed high in crude protein and fibre. Key words: Beta vulgaris L., agronomic practices, mineral concentration, mineral uptake, quality attributes.

New Zealand pastoral farming has benefited greatly from the application of phosphorus (P) and sulphur (S) fertilisers supplied in the main by superphosphate (SSP). The long-term fertiliser trial at Winchmore, mid-Canterbury, New Zealand was set up in 1952 and has yielded a wealth of data on the effect of fertiliser, grazing by sheep and flood irrigation on pasture production. The trial was initially (1952–1958) designed to measure the effect of no fertiliser and SSP applied each year at 188, 376 and 564 kg ha−1. From 1958 to 1980, SSP applications were stopped to the 564 and one half of the 376 kg SSP y−1. The cessation of fertiliser decreased clover content and increased the proportion of weeds and low-fertility grasses. The decline in production once fertiliser application ceased followed a curvilinear pattern, but never reached the low production of the no fertiliser treatments even after 20 years. In 1980, the residual treatments were changed to compare a reactive phosphate rock (RPR)/S treat...

Protein-based animal fibres of commercial importance are frequently exposed to elevated temperatures during processing treatments. Hydrothermal processes cause protein deterioration, impacting negatively on the value or condition of these materials. This study was designed to investigate hydrothermal damage in wool proteins at the molecular level. The effect of hydrothermal damage on Type I and II intermediate filament proteins (keratins) extracted from wool was characterised using advanced quantitative techniques based on isobaric iTRAQ labelling and mass spectrometry. Many native peptides were observed to be degraded and modified. Amongst these, twenty keratin peptides were observed to consistently degrade during hydrothermal exposure. These peptides acted as molecular markers of damage – specific indicators of the extent of heat-induced protein damage. This technology will be of value in assessing the severity of damage imparted after high temperature exposure of protein-based animal fibres such as wool and cashmere during processes such as dyeing and carbonising, or even after high temperature human hair treatments. The identification of molecular damage markers identified within wool and other materials provides a new route to sensitive and specific evaluation of the effects of protein deterioration. It is anticipated that the utilisation of such markers will facilitate the development of targeted approaches to minimising processing damage to high-value fibres and protein-based biomaterials.

Proteins from the plant chromoplast are essential for many physiological processes such as fatty acid biosynthesis. Different protein extraction methods were tested to find the most robust method to obtain oil palm chromoplast proteins for mass spectrometry analysis. Initially, two different solvents were employed to reduce the fruit lipids. Then, two plant cell wall digestive enzymes were used to acquire the protoplasts to increase the protein extraction effectiveness. A two-stage centrifugation-based fractionation approach enhanced the number of identified proteins, particularly the fatty acid biosynthetic enzymes. The effectiveness of each extraction method was assessed using protein yields and 2DE gel profiles. The ideal method was successfully used to establish the 2DE chromoplast proteome maps of low and high oleic acid mesocarps of oil palm. Further nanoLC–MS/MS analysis of the extracted chromoplast proteins led to the identification of 162 proteins, including some of the main enzymes involved in the fatty acid biosynthesis. The established procedures would provide a solid foundation for further functional studies, including fatty acid biosynthetic expression profiling and evaluation of regulatory function. Electronic supplementary material The online version of this article (doi:10.1186/s40064-015-1576-4) contains supplementary material, which is available to authorized users.

Field-trial data from a database comprising records of 804 potassium (K) fertiliser trials were used to define the production functions relating exchangeable soil K (quick test K (QTK) 0–75 mm) to the relative response to fertiliser K applications, for the major soil groups in New Zealand. For all soil groups for which there were sufficient data, the production functions were generally flat in the range QTK 5–10, and thus the estimated relative pasture production at QTK 5 and QTK 10 were similar. The critical QTK levels to achieve 97% maximum production were relatively well defined, being 6 (5–8) for sedimentary soils (brown and pallic) and brown soils, and 7 (5–10) for pumice soils. The data for the allophanic soils were unstable and the best estimate was 6 (5–10). For the remaining soils groups (podzols and raw soils, organic, recent and gley soils) for which there was much less data, the relationships were essentially flat over the range QTK 2–10. The probability of pasture responses to applie...

Thaumatin-like proteins (TLPs) and chitinases are the main constituents of so-called protein hazes which can form in finished white wine and which is a great concern of winemakers. These soluble pathogenesis-related (PR) proteins are extracted from grape berries. However, their distribution in different grape tissues is not well documented. In this study, proteins were first separately extracted from the skin, pulp and seed of Sauvignon Blanc grapes, followed by trypsin digestion and analysis by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Proteins identified included 75 proteins from Sauvignon Blanc grape skin, 63 from grape pulp and 35 from grape seed, mostly functionally classified as associated with metabolism and energy. Some were present exclusively in specific grape tissues; for example, proteins involved in photosynthesis were only detected in grape skin and proteins found in alcoholic fermentation were only detected in grape pulp. Moreover, proteins identified in grape seed were less diverse than those identified in grape skin and pulp. TLPs and chitinases were identified in both Sauvignon Blanc grape skin and pulp, but not in the seed. To relatively quantify the PR proteins, the protein extracts of grape tissues were seperated by HPLC first and then analysed by SDS-PAGE. The results showed that the protein fractions eluted at 9.3 min and 19.2 min under the chromatographic conditions of this study confirmed that these corresponded to TLPs and chitinases seperately. Thus, the relative quantification of TLPs and chitinases in protein extracts was carried out by comparing the area of corresponding peaks against the area of a thamautin standard. The results presented in this study clearly demonstrated the distribution of haze-forming PR proteins in grape berries, and the relative quantification of TLPs and chitinases could be applied in fast tracking of changes in PR proteins during grape growth and determination of PR proteins in berries at harvest.

Feeding out conserved hay or silage to animals over winter or in early spring is an integral part of New Zealand pastoral farming. However, the mineral content of conserved feed is usually below that of the original pasture, and is fed to stock at a time when their mineral requirements are high. During these periods, iodine and copper concentrations in pasture are marginal or deficient, exacerbating the problem. Three trials were laid down, two in Southland and one in the Waikato, looking at the uptake of copper (Cu), cobalt (Co), selenium (Se), and iodine (I) when applied in liquid form at various times between the shutting‐up stage and harvesting of conserved silage and hay paddocks. Application times for the Southland trials were 4 weeks prior to cutting, 2 weeks prior to cutting, the same day as cutting and onto the windrow after cutting. Minerals applied and rates of application were Cu: 0, 0.5, 1 and 3 kg copper sulphate ha–1, Se: 10 g Se ha–1 as sodium selenate, I:200 g potassium iodate ha...

A database was constructed comprising records from 2255 pasture phosphorus (P), potassium (K) and sulphur (S) field trials, of which 1799 included one or several rates of P. Subsets of this data were selected based on predetermined criteria to define the relationships between relative pasture production and available soil P (0–75 mm, Olsen P in μg P cm–3 soil)—the P production functions—for the major soil groups in New Zealand. These relationships, and their 95% confidence intervals, were defined using Bayesian statistics. For those soil groups for which there was sufficient data, the production functions were well defined and gave reasonably precise estimates of the relative pasture yield for a given Olsen P. For example, for the volcanic soils, the relative pasture production is most likely (P < 0.05) to be in the range 88–94% at Olsen P 25 and 98–100% at Olsen P 50. The shape of the production functions was similar for all soil groups—the relative pasture production increased with increasing O...

Modern processing techniques for turkey involve rapid chilling to slow microbial growth and early deboning of the economically important breast meat. This paper shows that these 2 processes lead to significantly tougher meat with higher cooking losses. The toughening appears to be due to less extensive proteolysis and shortening of the sarcomeres. Calpains I and II and their inhibitor, calpastatin, were quantified in turkey breast. Calpain II was the more common isoform but showed no evidence of activation during aging. In contrast, calpain I and calpastatin activities declined rapidly and were no longer detected 24 h postslaughter. There was no evidence of an association between calpain activity and processing conditions.

The effects of carnosine, rutin and quercetin on oxidative processes and metmyoglobin (MetMb)-reducing activity in a beef model system were investigated. Ground beef was mixed with antioxidant solution at two concentrations. Storage period and antioxidant treatment negatively affected colour, colour stability and thiobarbituric acid-reactive substances values. Carnosine, rutin and to lesser extent quercetin accelerated discolouration, accumulation of MetMb and lipid peroxidation in the meat. The results suggest that these antioxidants acted as a pro-oxidants under the specified experimental conditions. The pro-oxidant activities were in the following order: carnosine>rutin>quercetin. The MetMb-reducing activity was not correlated with storage time, colour parameters, MetMb (%) or lipid oxidation. The MetMb-reducing activity was significantly reduced by 1 mM carnosine. However, there was no association between MetMb-reducing activity and colour stability during post-mortem storage.

The influences of light and nitrogen (N) on growth and physiological characteristics of C. vitalba seedlings were investigated in a New Zealand lowland podocarp forest remnant. Seedlings planted within undisturbed forest did not persist, even when supplied with additional nutrients, whereas seedlings planted outside the forest not only persisted but, with additional nutrients, achieved substantial growth. Under controlled conditions with irradiance over a range from full sunlight (100% I r) to 1% full sunlight (1% I r), seedlings achieved maximum growth at 100% I r, substantial growth as low as 10% I r , and little growth at 3% I r. Seedlings at 1% I r did not survive. The influence of irradiance on seedling growth is compared with light acclimation characteristics; seedlings at low compared with high irradiances had a lower chlorophyll a:b ratio, lower concentration of total carotenoids and soluble protein per unit leaf area and chlorophyll, and a greater shoot:root (S:R) ratio, specific leaf ar...

The pattern of distribution of the invasive vine Clematis vitalba in a typical New Zealand native forest remnant and the influence of chilling duration (chilling), nitrate (NO3 -), and light on germination of C. vitalba were investigated. In the forest remnant, C. vitalba was not established in undisturbed forest where light levels were ≤3% fall sunlight and rarely established where light levels were ≤5%. Clematis vitalba was, however, established in forest gaps and margins where light levels exceeded 5%, particularly when these areas were associated with recent soil disturbance. Under controlled environment conditions, germination in the absence of chilling, NO3 -, or light was low (3%). When applied alone, chilling and NO3 - increased germination. Light, however, only increased germination when applied with one or both other factors. In the presence of light, germination increased up to c. 80% with increased chilling (0–12 weeks), and increased up to c. 50% with increased applied NO3 - in the r...

Effects of chilling (5 °C) period, light and applied nitrogen (N) on germination (%), rate of germination (d to 50% of total germination; T 50% ) and seed imbibition were examined in Clematis vitalba L. In the absence of chilling, light and N, germination was minimal (3%). When applied alone, both chilling and N increased germination. Chilling for 12 weeks increased germination to 64%, and 2.5 m M NO − 3 or NH + 4 increased germination to 10–12%. Light did not increase germination when applied alone, but did when applied in combination with chilling and/or N. Half the seed germinated when light was combined with 2.5 m M NO − 3 or NH + 4 . The influence of chilling, light and/or N on germination was greater when combined, than when either factor was applied alone. Both oxidized (NO − 3 ) and reduced (NH + 4 ) forms of N increased germination, but non-N-containing compounds did not, suggesting the response was due to N and not ionic or osmotic effects. Without additional N, T 50% decreased from 16–20 d at zero chilling, to around 5 d at 8 and 12 weeks chilling. Although T 50% was not influenced by an increase in NO − 3 or NH + 4 from 0.5 to 5.0 m M , it did increase with additional applied N thereafter. However, the magnitude of the N effect was small compared to that of chilling. Like germination, seed imbibition increased with a longer chilling period, but in contrast imbibition decreased slightly with increased applied NO − 3 or NH + 4 . It is argued that increased imbibition is not directly related to an increase in total germination, but that it may be related to the rate of germination. Possible mechanisms involved in the reduction in dormancy of C. vitalba seed are discussed.

Forty‐six data‐sets from a series of 17 long‐term field trials measuring the effects of rates and forms of phosphate (P) fertiliser on pasture production and soil fertility in New Zealand were used to characterise the relationship between pasture dry matter (DM) yield in any year (expressed as relative yield, RY) and soil Olsen P test at the end of the annual DM measurement period. Average coefficients of variation in the measurement of these parameters were 7% and 17% for DM yield and Olsen P, respectively. The results failed to demonstrate that the RY at any site in any year could be reliably assessed from a common relationship between RY and the Olsen P test. However, the mean relationship did take the general “diminishing returns” form as expressed by the Mitscherlich equation, and as Olsen P levels increased the probability of low RY decreased. There was no evidence that RY increased with increase in Olsen P test values above 20, with the exception of the one yellow‐brown pumice soil in the ...

North Otago soils are all of sedimentary origin but range in topography from flat alluvial and terrace soils to hill soils. Most of the farmed soils are yellow-grey earths on the rolling downlands or plains. Yellow-grey earths on the downlands have dense subsoils that limit water movement during wet winters and rooting depth during dry summers. Plains soils have shallow depths to gravels limiting water holding capacity and making irrigation necessary for intensive pastoral production. The relationships between soil nutrient levels and pasture production has been shown to be of the diminishing returns type. Soil nutrient levels for near maximum pasture production on North Otago soils are Olsen P 20-25, sulphate-S 10-12, organic-S 15-20 and quick test K 5-8. At high stocking rates profitable responses in pasture and animal production can still be gained at higher Olsen P levels. The decision support nutrient model (OUTLOOKTM) has been developed to help farmers apply fertiliser at optimum rates for economic production. Keywords: economic production, North Otago, phosphorus, potassium, sedimentary soils, sulphur, yellow-grey earths

We used sheep with an autosomal dominant gene for cortical cataract as an animal model to evaluate novel macrocyclic calpain inhibitors with potential for the medical treatment of human cataract.The macrocyclic aldehyde, CAT811, identified previously as a calpain inhibitor that prevents calcium-induced opacification in cultured sheep lenses, was tested for its ability to protect cytoskeletal proteins from calpain proteolysis. CAT811 and its alcohol analogue, CAT505, were formulated separately into ointments, and each was applied twice daily to the right eye of sheep with early cataracts for five months. Progress of cataracts in the sheep was determined by ophthalmologic examination and comparison with a matched sample of sheep treated similarly with ointment that did not contain the active ingredient.The novel macrocyclic aldehyde, CAT811, was able to inhibit calpain proteolysis of lens cytoskeletal proteins at micromolar concentrations. When applied topically to the eyes of sheep, CAT811 was able to slow cataract development by 27% in the initial three months of treatment (P0.05). Its alcohol analogue, CAT505, was not able to slow cataractogenesis significantly.The inherited sheep cataract provides a reproducible model of cortical cataract over a time scale of several months. The data reported here, using this model, demonstrated the potential of the macrocyclic calpain inhibitor, CAT811, to act as a therapeutic for treatment of cortical cataract.

The generally strongly weathered, leached soils of Northland consist of four major soil groups. The yellow-brown earths and podzols and yellow brown sands are formed from sedimentary rocks, while brown granular clays and red and brown loams are formed from volcanic rocks. In terms of the relationship between pasture production and fertiliser nutrient requirements, for both sedimentary and volcanic soils, the production functions are shown to be of the "diminishing returns" type, and the point at which near-maximum production (97%) occurs is defined as the "biological optimum" soil test level. Biological optimum test values for sedimentary and volcanic soils are: Olsen P 20 and 22; quicktest K 6 and 7; sulphate-S 10; organic-S 15; and pH 5.9. Once biological optimum soil test levels have been attained then maintenance fertiliser nutrient rates are appropriate. In order to move up the pasture production curve an average of 7 and 11 kg P/ha above maintenance will increase Olsen P by 1 unit for Northland sedimentary and volcanic soils respectively. Similarly, on average 60 kg K/ha will raise quicktest K by 1 unit on volcanic soils, but capital requirements for K on sedimentary soils in Northland are not known. An average of 35 and 25 kg S/ha will correct S deficiencies on sedimentary and volcanic soils. Keywords: biological optimum, lime, Northland, nutrient requirements, phosphorus, potassium, sedimentary soils, sulphur, volcanic soils

Purpose Cataracts are an important cause of blindness in humans but there are few large animal models available. One of these animal models is Ovine Heritable Cataract, a bilateral cortical cataract which develops after birth. This cataract has been used as a model for human cataracts in drug trials, but the gene responsible for the cataract trait is unknown. A genetic test for cataract would improve the efficiency of the model by predicting which animals would develop cataracts. Identifying the genetic basis of the cataract would indicate its relevance to human cataract. Methods A genome scan was performed on 20 sheep chromosomes, representing 86% of the genome, to determine the position of the cataract locus. Additional microsatellite markers were tested on chromosome 6 using a larger pedigree. Fine mapping was performed using a breakpoint panel of 36 animals and novel microsatellite markers taken from the bovine genome assembly. All exons of the candidate gene nudix (nucleoside diphosphate linked moiety X)-type motif 9 (NUDT9) were sequenced in normal and affected sheep. Results Significant linkage was found between cataract status and markers on chromosome 6. Linkage analysis on the larger pedigree showed the most likely position of the cataract locus was between 112.3 and 132.9 cM from the centromere. During fine mapping, NUDT9 was considered as a positional candidate for the cataract gene because it was located within the linked interval and is expressed in the lens. The gene was ruled out as the cataract gene after extensive genotype analysis, but a single nucleotide polymorphism (SNP) inside it provided a useful restriction fragment length polymorphism (RFLP) marker for further fine mapping. Twelve new markers were found and used to map the cataract locus to between 131.1 and 131.8 cM from the centromere. Conclusions A region of ovine chromosome 6 strongly linked to cataract has been identified, and a genetic test for cataract based on a SNP within this region has been developed. The best candidate gene within this region is AF4/FMR2 family, member 1 (AFF1), the mouse equivalent of which is associated with an inherited cataract.

Two new series of 15-membered macrocyclic peptidomimetics, in which the P1 and P3 residues of the peptide backbone are linked by a bridge containing a 1,4-disubstituted 1H-imidazole, are reported. The structure with an aldehyde at the C-terminus and the imidazole at P3, i.e., 4c, shows significant inhibitory activity against calpain 2, with an IC(50) value of 238 nM. The macrocyclic aldehyde with the imidazole at the alternative P1 position, i.e., 5c, is significantly less active. The relative activities are linked to the ability of the component macrocycles to mimic a β-strand geometry that is known to favor active-site binding. This ability is defined by conformational searches and docking studies with calpain.

The effects of annual applications at four different rates of either triple superphosphate (TSP) or Sechura phosphate rock (SPR) on the Olsen P test in soil under pasture were measured over 6 years at 13 sites in New Zealand. Averaged over the 13 sites, TSP applied at the calculated maintenance (M) rate maintained an almost constant Olsen P level throughout the 6 years. However, there were marked differences between sites, particularly at the 2.0 M application rate which at some sites raised Olsen P by over 30 μg P/ml soil but caused no increase at other sites. No obvious explanation such as soil group, phosphate retention, rainfall, or pH, or the TSP rates used could be found for these site differences. Olsen P tests were always markedly lower with SPR than with TSP; averaged over the 13 sites, even the 2.0 M SPR treatment barely maintained the initial Olsen P test values. This difference between TSP and SPR in their effect on Olsen P test contrasted with pasture production data (reported elsewh...

Pasture herbage dry matter (DM) responses to annual applications of triple superphosphate (TSP) at 0.5, 0.75, 1.0, and 2.0 times the calculated maintenance requirement were measured in 12 field trials for 6 years. The trials were on well‐established, permanent pastures which had typical fertiliser histories and represented major pastoral soils of North and South Islands of New Zealand. The DM response to TSP averaged over all rates and sites was 5% in Year 1, increasing to 23% in Year 4 with little change thereafter. At individual sites, average responses in total DM over 6 years ranged from 4 to 24%. Responses increased steadily with time at four sites, but at other sites changes with time were erratic. Mitscherlich curves were fitted to the 6‐year total DM production data for each site. However, the standard errors in the parameters of these fitted curves were very large, making it impossible to predict with any useful degree of accuracy the rate of TSP required for any specified yield. Respons...

The soils of North Canterbury occur mainly on steep land (31%), hilly land (28%), rolling downs and downlands (11%) and terraces and floodplains (30%). Most soils are predominantly formed from greywacke and tertiary sediments and some 85% comprise yellow-grey earth, yellow-grey earth to yellow-brown earth intergrade, yellow-brown earth and recent soil groups. In terms of the relationship between pasture production and fertiliser nutrient use these major soil groups are considered as a single entity, namely "sedimentary" soils (a reference to their derivation from sedimentary rocks). The relationships (production functions) between pasture production and soil test level for sulphur (S), phosphorus (P) and potassium (K) are shown to be of the "diminishing returns" type for the sedimentary soils and the point at which near-maximum production (97%) occurs is defined as the "biological optimum" soil test level. Biological optimum sulphate-S and organic-S levels-are-l.0 and 15 respectively, for O-l.s-en- P. -is _ 20 and quick-test K is 6. Once the biologrcal optimum soil test levels have been attained then maintenance fertiliser nutrient rates are appropriate. In order to correct S deficiency sedimentary soils require an average of 35 kg S/ha. To move up the pasture production curve an average of 5 kg P/ha above maintenance will increase Olsen P by 1 unit. Potassium requirements depend on soil group within the sedimentary soils. -Keywords:~biological~optimum, North Canterbury, nutrient requirements, phosphorus, potassium, sedimentary soils, Sulphur

Nitrate reductase activity (NRA, in vivo assay) and nitrate (NO 3 - ) content of root and shoot and NO 3 - and reduced nitrogen content of xylem sap were measured in five temperate cereals supplied with a range of NO 3 - concentrations (0.1-20 mol m -3 ) and three temperate pasture grasses supplied with 0.5 or 5.0 mol m -3 NO 3