Browsing by Author "Loveday SM"

Now showing 1 - 15 of 15
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    A protocol combining breath testing and ex vivo fermentations to study the human gut microbiome
    (Elsevier Inc, 2021-03-19) Payling L; Roy NC; Fraser K; Loveday SM; Sims IM; Janssen PH; Hill SJ; Raymond LG; McNabb WC
    This protocol describes the application of breath testing and ex vivo fermentations to study the association between breath methane and the composition and functionality of the gut microbiome. The protocol provides a useful systems biology approach for studying the gut microbiome in humans, which combines standardized methods in human breath testing and fecal sampling. The model described is accessible and easy to repeat, but its relative simplicity means that it can deviate from human physiological conditions.
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    Animal factors affecting the cheese-making properties and the heat coagulation time of milk from dairy sheep in a New Zealand flock
    (Taylor and Francis Group on behalf of the Royal Society of New Zealand, 2024-03-27) Marshall AC; Lopez-Villalobos N; Loveday SM; Weeks M; McNabb W
    The objective of this study was to evaluate the effect of animal factors on the cheese-making properties and on the heat coagulation time of milk from individual dairy sheep in a New Zealand flock. A total of 521 individual records were obtained from a seasonal pasture-based flock of 169 ewes milked once-a-day, from 50 to 182 days in milk. A statistical model was used to quantify the effects of animal factors (coat colour variety, age, litter size and stage of lactation) on the studied traits. Stage of lactation, confounded with seasonality, strongly influenced all properties of milk investigated. With the advancement of lactation, the milk took longer to coagulate after rennet addition, and the curd was softer. Higher relative cheese yield was achieved towards the end of lactation. The milk was also less stable to high-temperature treatment in late lactation. Coefficient of variation for processability traits was high and ranged from 20.2% to 58%, which can be largely attributed to stage of lactation but could also indicate room for genetic improvement of traits. Further genetic studies are underway to define animal genetic variance, heritability, and the phenotypic and genetic correlations between these processability and milk composition traits.
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    Digestive diversity and kinetic intrigue among heated and unheated β-lactoglobulin species.
    (ROYAL SOC CHEMISTRY, 2014-11) Loveday SM; Peram MR; Singh H; Ye A; Jameson GB
    Food processing often alters the structure of proteins, and proteins are deliberately denatured and aggregated to improve technological functionality in many cases. However, the digestive consequences of processing-induced alterations to protein structure have only recently been studied. Here we explored the process-structure-digestibility relationship in the context of heat-processing effects on the structure and gastric digestibility of the bovine whey protein β-lactoglobulin (β-lg). Heating β-lg produces an array of non-native monomers, dimers and aggregates, and we have characterised these with reverse-phase high performance liquid chromatography (RP-HPLC) as a complement to our earlier work using polyacrylamide gel electrophoresis (PAGE) techniques. Using a combination of SDS-PAGE and RP-HPLC we have identified pepsin-resistant dimers and peptides that appear early in digestion. In an unexpected finding, native β-lg underwent complete hydrolysis during prolonged incubation (48 h) with pepsin. Two phases of hydrolysis were identified, and the transition between phases appears to result from alterations to the secondary structure of β-lg at 3-4 h, as measured with circular dichroism spectroscopy, and/or the binding and release of a pepsin inhibitor peptide. This work has unpacked some of the complexities of the processing-structure-digestibility relationship in a highly simplified system; further work is needed to explore the implications of these findings for food processors, regulatory authorities and consumers.
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    Effect of protein polymorphisms on milk composition, coagulation properties, and protein profile in dairy sheep
    (Elsevier Ltd, 2025-01-01) Marshall AC; Vigolo V; De Marchi M; Lopez-Villalobos N; Loveday SM; Weeks M; McNabb W
    The objective of this study was to quantify the effect of protein polymorphisms on milk composition, coagulation properties, and protein profile in dairy sheep from a New Zealand flock. A total of 470 test-day records, from 147 lactating ewes, were used in the statistical analyses. Protein polymorphisms observed in the RP-HPLC were self-named for purposes of the present study. Data were analyzed using a mixed linear model, including the fixed effects of ewe age, litter size, coat-colour, and stage of lactation, and, as a covariate, deviation from the median lambing date of the flock. The effects of protein polymorphisms were added to the model, one at a time. Protein polymorphisms were significantly (p < 0.05) associated with milk composition and protein profile. Polymorphisms of β-lactoglobulin were significantly associated with milk heat stability, being AB type more heat stable than AA. The other processability traits were not significantly affected by protein polymorphisms. Further studies are required to confirm the protein variants and the properties of individual protein polymorphisms.
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    Estimation of genetic parameters for production, composition and processability of milk from dairy sheep in a New Zealand flock
    (Taylor and Francis Group, 2024-06-27) Marshall AC; Lopez-Villalobos N; Loveday SM; Weeks M; McNabb W
    This paper aimed to estimate the heritability and genetic correlations for processability (milk coagulation properties, cheese yield, and heat stability), milk production, and milk composition of dairy sheep in a New Zealand flock. Test-day records from 169 ewes were obtained during the 2021–2022 season. Heritability estimates for yield traits (milk, fat, protein, and lactose yield) were moderate and ranged from 0.33 to 0.38. Whereas heritability estimates for processability traits were lower and ranged from 0.17 to 0.27. The genetic correlations indicated that poorer milk coagulation properties were associated with lower mammary health indicated by higher SCS, higher pH, and lower lactose content. Additionally, a higher ratio of casein to calcium i.e. lower availability of soluble calcium was associated with poorer milk coagulation properties. Higher yields of milk, fat, protein, and lactose were associated, but to a smaller degree, with better MCP. It is proposed that genetic improvement of dairy sheep for higher yields of fat and protein, and for lower SCS, could indirectly improve milk processability for cheese-making. However, it is recommended that larger studies involving more dairy sheep flocks are conducted to validate the present results before developing a selection index for this industry.
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    Genome-wide association studies for citric and lactic acids in dairy sheep milk in a New Zealand flock
    (Taylor and Francis Group, 2024-08-05) Zongqi A; Marshall AC; Jayawardana JMDR; Weeks M; Loveday SM; McNabb W; Lopez-Villalobos N
    The objectives of this study were to estimate genetic parameters for citric acid content (CA) and lactic acid content (LA) in sheep milk and to identify the associated candidate genes in a New Zealand dairy sheep flock. Records from 165 ewes were used. Heritability estimates based on pedigree records for CA and LA were 0.65 and 0.33, respectively. The genetic and phenotypic correlations between CA and LA were strong-moderate and negative. Estimates of genomic heritability for CA and LA were also high (0.85, 0.51) and the genomic correlation between CA and LA was strongly negative (-0.96 ± 0.11). No significant associations were found at the Bonferroni level. However, one intragenic SNP in C1QTNF1 (chromosome 11) was associated with CA, at the chromosomal significance threshold. Another SNP associated with CA was intergenic (chromosome 15). For LA, the most notable SNP was intragenic in CYTH1 (chromosome 11), the other two SNPs were intragenic in MGAT5B and TIMP2 (chromosome 11), and four SNPs were intergenic (chromosomes 1 and 24). The functions of candidate genes indicate that CA and LA could potentially be used as biomarkers for energy balance and clinical mastitis. Further research is recommended to validate the present results.
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    Heat-induced colloidal interactions of whey proteins, sodium caseinate and gum arabic in binary and ternary mixtures
    (Elsevier Ltd, 2013-11) Loveday SM; Ye A; Anema SG; Singh H
    Many food-grade proteins and polysaccharides will aggregate together when acidified or heated, due to electrostatic and hydrophobic interactions. At low concentrations, aggregates are soluble and colloidally stable, and they have potential applications as Pickering emulsifiers and nutrient carriers. Sodium caseinate (SC) and gum arabic (GA) at pH. 7 will form colloidal aggregates when heated, but aggregation is largely reversed on cooling. Whey proteins (in the form of whey protein isolate, WPI) will aggregate irreversibly with GA when they are heated together, but aggregation is often so rapid and extensive that aggregates precipitate. Here we sought to overcome those limitations, and to develop an in situ method for quantifying heat-induced aggregation. Aggregation was measured using temperature-controlled dynamic light scattering equipment and transmission electron microscopy. Combinations of SC, WPI and GA were heated at either pH. 7 or 3.5, and the weight ratio of protein to polysaccharide was held at 1:5 for simplicity. Heat-induced colloidally stable aggregates of SC. +. WPI. +. GA did not dissociate on cooling. Aggregation was measured in situ, both in temperature ramps and with isothermal experiments. In situ measurement allowed us to avoid potential artefacts stemming from the temperature changes and measurement delays associated with ex situ measurements. This work demonstrated how the size and heat-stability of colloidal protein-polysaccharide aggregates can be tailored by judicious selection of proteins, pH and heat treatment.
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    Heat-Treatments Affect Protease Activities and Peptide Profiles of Ruminants' Milk
    (Frontiers Media S.A., 2021-03-10) Leite JAS; Montoya CA; Loveday SM; Maes E; Mullaney JA; McNabb WC; Roy NC; Abd El-Aty, AM
    Proteases present in milk are heat-sensitive, and their activities increase or decrease depending on the intensity of the thermal treatment applied. The thermal effects on the protease activity are well-known for bovine milk but poorly understood for ovine and caprine milk. This study aimed to determine the non-specific and specific protease activities in casein and whey fractions isolated from raw bovine, ovine, and caprine milk collected in early lactation, and to determine the effects of low-temperature, long-time (63°C for 30 min) and high-temperature, short-time (85°C for 5 min) treatments on protease activities within each milk fraction. The non-specific protease activities in raw and heat-treated milk samples were determined using the substrate azocasein. Plasmin (the main protease in milk) and plasminogen-derived activities were determined using the chromogenic substrate S-2251 (D-Val-Leu-Lys-pNA dihydrochloride). Peptides were characterized using high-resolution liquid chromatography coupled with tandem mass spectrometry. The activity of all native proteases, shown as non-specific proteases, was similar between raw bovine and caprine milk samples, but lower (P < 0.05) than raw ovine milk in the whey fraction. There was no difference (P > 0.05) between the non-specific protease activity of the casein fraction of raw bovine and caprine milk samples; both had higher activity than ovine milk. After 63°C/30 min, the non-specific protease activity decreased (44%; P > 0.05) for the bovine casein fraction only. In contrast, the protease activity of the milk heated at 85°C/5 min changed depending on the species and fraction. For instance, the activity decreased by 49% for ovine whey fraction, but it increased by 68% for ovine casein fraction. Plasmin and plasminogen were in general inactivated (P > 0.05) when all milk fractions were heated at 85°C/5 min. Most of the peptides present in heat-treated milk were derived from β-casein and αS1-casein, and they matched the hydrolysis profile of cathepsin D and plasmin. Identified peptides in ruminant milk samples had purported immunomodulatory and inhibitory functions. These findings indicate that the non-specific protease activity in whey and casein fractions differed between ruminant milk species, and specific thermal treatments could be used to retain better protease activity for all ruminant milk species.
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    Intragastric restructuring dictates the digestive kinetics of heat-set milk protein gels of contrasting textures
    (Elsevier, 2024-11) Li S; Mungure T; Ye A; Loveday SM; Ellis A; Weeks M; Singh H
    The gelation of milk proteins can be achieved by various means, enabling the development of diverse products. In this study, heat-set milk protein gels (15 % protein) of diverse textures were made by pH modulation and two gels were selected for dynamic in vitro gastric digestion: a spoonable soft gel (SG, pH 6.55' G' of ∼100 Pa) and a sliceable firm gel (FG, pH 5.65; G' of ∼7000 Pa). The two gels displayed markedly different structural changes and digestion kinetics during gastric digestion. The SG underwent substantial structural compaction during the first 120 min of gastric digestion into a denser and firmer gastric chyme (26.3 % crude protein, G* of ∼8500 Pa) than the chyme of the FG (15.7 % crude protein, G* of ∼3000 Pa). These contrasting intragastric structural changes of the gels reversed their original textural differences, which led to slower digestion and gastric emptying of proteins from the SG compared with the FG. The different intragastric pH profiles during the digestion of the two gels likely played a key role by modulating the proteolytic activity and specificity (to κ-casein) of pepsin. Preferential early cleavage of κ-casein in SG stimulated coagulation and compaction of solid chyme, whereas rapid hydrolysis of αS- and β-caseins in the FG weakened coagulation. This study provided new insights into controlling the structural development of dairy-based foods during gastric digestion and modulating digestion kinetics.
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    Modelling Lactation Curves for Dairy Sheep in a New Zealand Flock
    (MDPI (Basel, Switzerland), 2023-01-19) Marshall AC; Lopez-Villalobos N; Loveday SM; Ellis A; McNabb W
    Lactation curves were modelled for dairy sheep in a New Zealand flock, providing information on the lactation yields of milk, fat, protein, and lactose, corrected for 130 days of milking. From 169 ewes, a total of 622 test-day records were obtained during the milk production season of 2021-2022 (from October to January). The flock produced an average of 86.1 kg of milk, 5.1 kg of fat, 4.5 kg of protein, and 4.1 kg of lactose, and moderate to large coefficients of variation were observed (27-31%) for these traits. The lactation persistency of milk, fat, protein, and lactose yields ranged from 52.3 to 72.7%. Analyses of variance for total yield and persistency were performed with an animal model that included the fixed effects of age (parity number), litter size, coat colour, and milking frequency (days in twice-a-day milking) and random residuals. Age and milking frequency were the only factors that significantly affected the yields of milk, fat, protein, and lactose. Age significantly affected the lactation persistency of milk and lactose yields, whereas litter size affected the persistency of protein, and milking frequency affected the persistency of fat. This study on this single flock provides valuable experience for a larger-scale animal breeding programme in New Zealand.
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    Soy Protein Pressed Gels: Gelation Mechanism Affects the In Vitro Proteolysis and Bioaccessibility of Added Phenolic Acids
    (MDPI (Basel, Switzerland), 13/01/2021) Marinea M; Ellis A; Golding M; Loveday SM
    In this study, a model system of firm tofu (pressed gel) was prepared to study how the coagulation mechanism-acidification with glucono δ-lactone (GDL) or coagulation with magnesium sulphate (MgSO4)-affected the physical properties of the gels along with their in vitro proteolysis (or extent of proteolysis). The two types of gels were also fortified with 3.5 mM protocatechuic (PCA) and coumaric acid (CMA) to test whether they can be used as bioactive delivery systems. Texture analysis showed that all MgSO4-induced gels (fortified and control) had a higher hydration capacity and a weaker texture than the GDL-induced gels (p < 0.05). MgSO4 gels had almost double proteolysis percentages throughout the in vitro digestion and showed a significantly higher amino acid bioaccessibility than the GDL gels (essential amino acid bioaccessibility of 56% versus 31%; p < 0.05). Lastly, both gel matrices showed a similar phenolic acid release profile, on a percentage basis (~80% for PCA and ~100% for CMA). However, GDL gels delivered significantly higher masses of bioactives under simulated intestinal conditions because they could retain more of the bioactives in the gel after pressing. It was concluded that the coagulation mechanism affects both the macro- and microstructure of the soy protein pressed gels and as a result their protein digestibility. Both pressed gel matrices are promising delivery systems for bioactive phenolic acids.
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    The impact of heating and drying on protease activities of ruminant milk before and after in vitro infant digestion
    (Elsevier Ltd, 2023-12-15) Leite JAS; Montoya CA; Loveday SM; Mullaney JA; Loo TS; McNabb WC; Roy NC
    This study investigated the effect of heating (63°C/30 min or 75°C/15 s) and drying (spray-drying or freeze-drying) on plasmin, cathepsin D, and elastase activities in bovine, ovine, and caprine milk, compared to non-dried raw milk counterparts. Protease activities and protein hydrolysis were assessed before and after in vitro infant digestion with or without gastric and pancreatic enzymes. At 75°C/15 s, plasmin activity in caprine and ovine milk decreased (69-75%, p<0.05), while cathepsin D activity in spray-dried bovine milk heated increased (2.8-fold, p<0.05). Plasmin and cathepsin D activities increased (<1.2-fold, p<0.05) after in vitro digestion with pancreatin, regardless of milk species. Endogenous milk enzymes hydrolyzed more proteins than gastric enzymes during gastric digestion and contributed to small intestinal digestion. In summary, milk proteases remained active after processing with effects dependent on the species of milk, and they contributed to in vitro protein hydrolysis in the stomach and small intestine.
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    The use of confocal Raman microscopy and microfluidic channels to monitor the location and mobility of β-carotene incorporated in droplet-stabilized oil-in-water emulsions
    (Elsevier B V, 2023-05-17) Okubanjo SS; Brooke SJ; Ward R; Mostert N; Loveday SM; Ye A; Wilde PJ; Singh H; Waterland M
    This study sought to explore the combined use of confocal Raman microscopy and microfluidic channels to probe the location and mobility of hydrophobic antioxidant (β-carotene) incorporated at the interface of food-grade droplet-stabilized emulsions (DSEs). Microfluidic channels were used to isolate emulsion droplets for efficient investigation of antioxidant mobility. This approach proved more conclusive than fixing the sample in agarose, because a single layer of droplets could be obtained. Results also indicated that the migration of β-carotene incorporated in shell droplets of olive oil and trimyristin DSEs to core droplets was minimal and beta-carotene remained mostly localised at the interface even after 3 days of production. This work demonstrates that microfluidic isolation of emulsion droplets combined with confocal Raman microscopy can give new insights into the spatial variation of chemical composition within emulsions. This study revealed that the migration of β-carotene between shell and core was minimal and hence it may be possible to concurrently deliver two incompatible compounds by spatially segregating them between shell and core compartments of DSEs.
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    Udder and teat morphology traits associated with milk production and somatic cell score in dairy sheep from a New Zealand flock
    (Taylor and Francis Group on behalf of the Royal Society of New Zealand, 2024) Marshall AC; Lopez-Villalobos N; Loveday SM; Weeks M; McNabb W; Handcock R
    The objective of this study was to estimate the phenotypic correlations between udder and teat morphology traits, milk production traits, and somatic cell score in dairy sheep from a flock in New Zealand. A total of 162 lactating ewes were scored for morphology traits during the milk production season of 2021–2022. The 130-d lactation yields of milk, fat, protein, and lactose were obtained with 2–4 test-days from each ewe and modelled using random regression with orthogonal polynomials. Age had a significant effect on all udder and teat traits. Coat-colour (genetic variety within the breed; white or black) was a significant effect for teat angle and udder separation. Udders that were above the hook were associated with lower milk, fat, protein, and lactose yields. Udders with well-defined separation between halves were associated with higher milk, protein, and lactose yields, and with lower somatic cell count. Well-attached udders were associated with lower fat yield and lower somatic cell score. Teats with a backwards angle were associated with lower milk and lactose yields. Further studies are needed to estimate heritability and genetic correlations between these traits to determine whether these traits should be implemented in breeding programs for dairy sheep in New Zealand.
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    β-Lactoglobulin nanofibrils: Effect of temperature on fibril formation kinetics, fibril morphology and the rheological properties of fibril dispersions
    (Elsevier Ltd, 2012-05) Loveday SM; Wang XL; Rao MA; Anema SG; Singh H
    Almost all published studies of heat-induced b-lactoglobulin self-assembly into amyloid-like fibrils at low pH and low ionic strength have involved heating at 80 C, and the effect of heating temperature on self-assembly has received little attention. Here we heated b-lactoglobulin at pH 2 and 75 C, 80 C, 90 C, 100 C, 110 C or 120 C and investigated the kinetics of self-assembly (using Thioflavin T fluorescence), the morphology of fibrils, and the rheological properties of fibril dispersions. Self-assembly occurred at all temperatures tested. Thioflavin T fluorescence increased sigmoidally at all temperatures, however it decreased sharply with >3.3 h heating at 110 C and with >5 h heating at 120 C. The sharp decreases were attributed partly to local gelation, but destruction of fibrils may have occurred at 120 C. Thioflavin T fluorescence results indicated that maximal rates of fibril formation increased with increasing temperature, especially above 100 C, but fibril yield (maximum Thioflavin T fluorescence) was not affected by temperature. At 100 C and 110 C, fibrils were slightly shorter than at 80 C, but otherwise they looked very similar. Fibrils made by heating at 120 C for 1 h were also similar, but heating at 120 C for 8 h gave predominantly short fibrils, apparently the products of larger fibrils fragmenting. Heating at 100 C gave consistently higher viscosity than at 80 C, and heating for >2 h at 120 C decreased viscosity, which may have been linked with fibril fragmentation seen in micrographs.