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Journal = Dairy
Section = Dairy Systems Biology

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28 pages, 4968 KiB  
Article
Peptide Profiling in Dairy Cow Dry Secretions: Temporal Changes and Comparative Analysis Between Healthy and Subclinical Mastitis Cows
by Barjam Hasanllari, Benjamin P. Willing, Liang Li, Xian Luo and Burim N. Ametaj
Dairy 2025, 6(2), 19; https://doi.org/10.3390/dairy6020019 - 15 Apr 2025
Viewed by 1162
Abstract
The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we [...] Read more.
The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we employed peptidomics to explore the peptide profiles of dry secretions collected from dairy cows at specific intervals during the dry-off period. We hypothesized that alterations in peptide composition during the dry period may influence pathogen proliferation and immune cell functioning, thereby impacting mastitis susceptibility. Our objectives were to investigate the following: (i) differences in peptide composition and alterations between healthy cows and those with subclinical mastitis, potentially serving as biomarkers for early mastitis detection and offering insights into udder bioprocesses; (ii) variations in peptide profiles between the early (day 2) and later (day 21) stages of the dry-off period during both health conditions. Dry secretions were collected from 16 udder quarters of 8 cows at two defined time points—Day 2 (D2) and Day 21 (D21)—during the dry period. Our results revealed distinct peptide patterns between healthy and subclinical mastitis cows, as well as temporal variations in peptide profiles throughout the dry-off period. A total of 1235 peptides, originating from 59 distinct proteins (primarily β-casein), were detected across the four groups: subclinical mastitis day 21 (SCM-D21), subclinical mastitis day 2 (SCM-D2), healthy day 21 (H-D21), and healthy day 2 (H-D2). Furthermore, 56 out of the 1235 peptides identified in total matched known functional peptides, with a total of 17 different functions including antimicrobial, antioxidant, and immunomodulatory, suggesting their potential roles in mastitis pathogenesis and mammary gland physiology. Comparative analyses revealed changes in the levels of these functional peptides across the four different groups, suggesting their potential roles in regulating immune responses, oxidative stress, inflammatory processes, and other biological activities during subclinical mastitis and the dry-off period. These findings provide valuable insights into mastitis detection, management strategies, and dairy cow health monitoring, offering promising avenues for enhancing milk quality and dairy industry sustainability. Full article
(This article belongs to the Section Dairy Systems Biology)
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14 pages, 2230 KiB  
Article
Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
by Leandra Oliveira Xavier Albiero, Rafaela Ansiliero and Aniela Pinto Kempka
Dairy 2024, 5(4), 786-799; https://doi.org/10.3390/dairy5040057 - 9 Dec 2024
Cited by 2 | Viewed by 2467
Abstract
In this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test [...] Read more.
In this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test was developed to identify β-casein A1 in fermented milk, and a commercial Lateral Flow Immunoassay (LFIA) was evaluated to determine A2A2 genotypes and the purity of A2 milk. The results demonstrated 100% sensitivity and specificity of the ELISA test in identifying β-casein A1. The LFIA test successfully identified A2A2 genotypes and confirmed the purity of A2 milk, with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Both tests exhibited 100% sensitivity and specificity, resulting in positive and negative predictive values of 100%. The positive likelihood ratio was infinite, while the negative was zero, indicating a precise and reliable test with no false diagnoses. Compared to traditional genotyping, these methods proved to be more practical and showed potential for large-scale screening. It was concluded that ELISA and LFIA are valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products. Full article
(This article belongs to the Section Dairy Systems Biology)
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29 pages, 550 KiB  
Review
Listeria monocytogenes: An Inconvenient Hurdle for the Dairy Industry
by Alessandra Casagrande Ribeiro, Felipe Alves de Almeida, Mariana Medina Medeiros, Bruna Ribeiro Miranda, Uelinton Manoel Pinto and Virgínia Farias Alves
Dairy 2023, 4(2), 316-344; https://doi.org/10.3390/dairy4020022 - 19 Apr 2023
Cited by 32 | Viewed by 11038
Abstract
Listeria monocytogenes is an opportunistic pathogen that affects specific groups of individuals, with a high mortality rate. The control of L. monocytogenes in dairy industries presents particular challenges, as this bacterium is capable of adhering and forming biofilms, as well as thriving at [...] Read more.
Listeria monocytogenes is an opportunistic pathogen that affects specific groups of individuals, with a high mortality rate. The control of L. monocytogenes in dairy industries presents particular challenges, as this bacterium is capable of adhering and forming biofilms, as well as thriving at refrigerated temperatures, which enables it to persist in harsh environments. The consumption of dairy products has been linked to sporadic cases and outbreaks of listeriosis, and L. monocytogenes is frequently detected in these products in retail stores. Moreover, the bacterium has been shown to persist in dairy-processing environments. In this work, we review the main characteristics of L. monocytogenes and listeriosis, and highlight the factors that support its persistence in processing environments and dairy products. We also discuss the main dairy products involved in outbreaks of listeriosis since the early 1980s, and present control measures that can help to prevent the occurrence of this pathogen in foods and food-processing environments. Full article
(This article belongs to the Section Dairy Systems Biology)
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16 pages, 989 KiB  
Article
Fatty Acid, Carotenoid and Fat-Soluble Vitamin Composition of Multispecies Swards Grown in Ireland—Implications for a Sustainable Feed in Dairy Farming
by Samuel Rapisarda, Graham O’Neill and Nissreen Abu-Ghannam
Dairy 2023, 4(2), 300-315; https://doi.org/10.3390/dairy4020021 - 17 Apr 2023
Cited by 4 | Viewed by 3055
Abstract
Although traditional ryegrass pastures are still commonly used, they require intense management and input and do not perform well during the summer months. Multispecies systems have been recognised as more sustainable, needing less fertiliser and being tolerant to drought and heat. While data [...] Read more.
Although traditional ryegrass pastures are still commonly used, they require intense management and input and do not perform well during the summer months. Multispecies systems have been recognised as more sustainable, needing less fertiliser and being tolerant to drought and heat. While data on monoculture system fat composition exist, information for multispecies is scarce. The present study compared the fatty acid (FA), carotenoid and fat-soluble composition of a multispecies system (consisting of perennial ryegrass, timothy, white clover, red clover, chicory and plantain) (MULTI) with two other conventional grazing systems (monoculture perennial ryegrass (PRG) and a binary mixture of perennial ryegrass and white clover (PRG+WC)) over the Irish late-summer grazing season of dairy cattle (July–September). The results showed that the three systems had similar levels of FAs (p > 0.05), with mean α-linolenic acid values of 20.00 mg/g in PRG, 18.51 mg/g in PRG+WC, and 17.90 mg/g in MULTI and mean linoleic acid values of 3.84 mg/g in PRG, 4.16 mg/g in PRG+WC, and 4.39 mg/g in MULTI. Fourfold and twofold increases in the concentrations of α-linolenic acid and linoleic acid, respectively, were observed throughout July–September. Variations in stearic acid concentrations were not significant (p > 0.05), and palmitic acid concentrations increased over time (p < 0.05). The average values of lutein (36.68 mg/kg in PRG, 31.26 mg/kg in PRG+WC, and 35.75 mg/kg in MULTI) and α-tocopherol (2.28 mg/kg in PRG, 1.49 mg/kg in PRG+WC, and 1.83 mg/kg in MULTI) were similar among the grazing systems (p > 0.05), and the average β-carotene value was 25% higher in monoculture ryegrass (140.74 mg/kg) than in the multispecies (102.51 mg/kg) (p < 0.05). Nonetheless, the levels of β-carotene found in the multispecies were still above the recommended intake required for ruminants. In conclusion, multispecies pastures could be included in dairy cattle production systems as a sustainable alternative to traditional pastures while also providing typically adequate sources of fats for animal diets. Full article
(This article belongs to the Special Issue Environmental Sustainability of Dairy Animal Systems)
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17 pages, 3559 KiB  
Article
The Impact of Low-Temperature Inactivation of Protease AprX from Pseudomonas on Its Proteolytic Capacity and Specificity: A Peptidomic Study
by Chunyue Zhang, Sjef Boeren, Liming Zhao, Etske Bijl and Kasper Hettinga
Dairy 2023, 4(1), 150-166; https://doi.org/10.3390/dairy4010011 - 3 Feb 2023
Cited by 4 | Viewed by 3195
Abstract
The destabilization of UHT milk during its shelf life can be promoted by the residual proteolytic activity attributed to the protease AprX from Pseudomonas. To better understand the hydrolysis patterns of AprX, and to evaluate the feasibility of using low-temperature inactivation (LTI) [...] Read more.
The destabilization of UHT milk during its shelf life can be promoted by the residual proteolytic activity attributed to the protease AprX from Pseudomonas. To better understand the hydrolysis patterns of AprX, and to evaluate the feasibility of using low-temperature inactivation (LTI) for AprX, the release of peptides through AprX activity on milk proteins was examined using an LC-MS/MS-based peptidomic analysis. Milk samples were either directly incubated to be hydrolyzed by AprX, or preheated under LTI conditions (60 °C for 15 min) and then incubated. Peptides and parent proteins (the proteins from which the peptides originated) were identified and quantified. The peptides were mapped and the cleavage frequency of amino acids in the P1/P1′ positions was analyzed, after which the influence of LTI and the potential bitterness of the formed peptides were determined. Our results showed that a total of 2488 peptides were identified from 48 parent proteins, with the most abundant peptides originating from κ-casein and β-casein. AprX may also non-specifically hydrolyze other proteins in milk. Except for decreasing the bitterness potential in skim UHT milk, LTI did not significantly reduce the AprX-induced hydrolysis of milk proteins. Therefore, the inactivation of AprX by LTI may not be feasible in UHT milk production. Full article
(This article belongs to the Special Issue Omics Research in Dairy Sciences)
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26 pages, 721 KiB  
Review
Adverse Effects of Fusarium Toxins in Ruminants: A Review of In Vivo and In Vitro Studies
by Antonio Gallo, Martina Mosconi, Erminio Trevisi and Regiane R. Santos
Dairy 2022, 3(3), 474-499; https://doi.org/10.3390/dairy3030035 - 11 Jul 2022
Cited by 20 | Viewed by 5030
Abstract
With an increased knowledge of the mechanism of action of Fusarium mycotoxins, the concept that these substances are deleterious only for monogastric species is obsolete. Indeed, most mycotoxins can be converted into less toxic compounds by the rumen microflora from healthy animals. However, [...] Read more.
With an increased knowledge of the mechanism of action of Fusarium mycotoxins, the concept that these substances are deleterious only for monogastric species is obsolete. Indeed, most mycotoxins can be converted into less toxic compounds by the rumen microflora from healthy animals. However, mycotoxin absorption and its conversion to more toxic metabolites, as well as their impact on the immune response and subsequently animal welfare, reproductive function, and milk quality during chronic exposure should not be neglected. Among the Fusarium mycotoxins, the most studied are deoxynivalenol (DON), zearalenone (ZEN), and fumonisins from the B class (FBs). It is remarkable that there is a paucity of in vivo research, with a low number of studies on nutrient digestibility and rumen function. Most of the in vitro studies are related to the reproductive function or are restricted to rumen incubation. When evaluating the production performance, milk yield is used as an evaluated parameter, but its quality for cheese production is often overlooked. In the present review, we summarize the most recent findings regarding the adverse effects of these mycotoxins with special attention to dairy cattle. Full article
(This article belongs to the Special Issue Effect of Mycotoxins on Dairy Production and Reproduction)
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12 pages, 1563 KiB  
Article
First Insight into the Variation of the Milk Serum Proteome within and between Individual Cows
by Lina Zhang, Sjef Boeren, Jeroen Heck, Jacques Vervoort, Peng Zhou and Kasper Hettinga
Dairy 2022, 3(1), 47-58; https://doi.org/10.3390/dairy3010004 - 14 Jan 2022
Cited by 1 | Viewed by 3047
Abstract
Milk contains all nutrients needed for development of calves. One important group of components responsible for this are the milk proteins. Variation due to feed or animal health, has been studied for the most abundant milk proteins. The aim of this study was [...] Read more.
Milk contains all nutrients needed for development of calves. One important group of components responsible for this are the milk proteins. Variation due to feed or animal health, has been studied for the most abundant milk proteins. The aim of this study was to determine the variation between and within cows for their milk serum proteome. Sample Set 1 was collected from Holstein Friesian (HF) cows between November 2011 and March 2012 and prepared using filter aided sample preparation (FASP) followed by LC-MS/MS for protein identification and quantification. The results showed that the milk serum proteome was very constant in mid lactation (four cows at five time points, p > 0.05) between 3 and 6 months in lactation. Sample Set 2 was collected from HF cows in Dec 2012 and analyzed using FASP and dimethyl labeling followed by LC-MS/MS. Significant variation in the milk serum proteome (p < 0.05) between 17 individual cows was found in Sample Set 2. The most variable proteins were immune-related proteins, which may reflect the health status of the individual cow. On the other hand, proteins related to nutrient synthesis and transport were relatively constant, indicating the importance of milk in providing a stable supply of nutrients to the neonate. In conclusion, the milk serum proteome was stable over mid lactation, but differed significantly between individuals, especially in immune-related proteins. Full article
(This article belongs to the Section Dairy Systems Biology)
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9 pages, 5696 KiB  
Article
Insights from Initial Variant Detection by Sequencing Single Sperm in Cattle
by Liu Yang, Yahui Gao, Clarissa Boschiero, Li Li, Hongping Zhang, Li Ma and George E. Liu
Dairy 2021, 2(4), 649-657; https://doi.org/10.3390/dairy2040050 - 15 Nov 2021
Cited by 1 | Viewed by 3694
Abstract
Meiotic de novo mutation (DNM) is one of the important phenomena contributing to gamete genome diversity. However, except for humans and a few model organisms, they are not well studied in livestock, including cattle. Moreover, bulk sperm samples have been routinely utilized in [...] Read more.
Meiotic de novo mutation (DNM) is one of the important phenomena contributing to gamete genome diversity. However, except for humans and a few model organisms, they are not well studied in livestock, including cattle. Moreover, bulk sperm samples have been routinely utilized in experiments, which include millions of single sperm cells and only report high-frequency variants. In this study, we isolated and sequenced 143 single sperms from two Holstein bulls and identified hundreds of candidate DNM events in ten sperms with deep sequencing coverage. We estimated DNM rates ranging from 1.08 × 10−8 to 3.78 × 10−8 per nucleotide per generation. We further validated 12 out of 14 selected DNM events using Sanger sequencing. To our knowledge, this is the first single sperm whole-genome sequencing effort in livestock, which provided useful information for future studies of point mutations and male fertility. Our preliminary results pointed out future research directions and highlighted the importance of uniform whole genome amplification, deep sequence coverage, and dedicated software pipelines for genetic variant detection using single-cell sequencing data. Full article
(This article belongs to the Section Dairy Systems Biology)
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13 pages, 1199 KiB  
Article
Influence of Dry Period Length of Swedish Dairy Cows on the Proteome of Colostrum
by Ruben de Vries, Sjef Boeren, Kjell Holtenius, Jacques Vervoort, Helena Lindmark-Månsson and Kasper Hettinga
Dairy 2020, 1(3), 313-325; https://doi.org/10.3390/dairy1030021 - 11 Dec 2020
Viewed by 3178
Abstract
The aim of this study was to evaluate the influence of applying a 4-week instead of an 8-week dry period to dairy cows on the proteome of colostrum (first sample) and of transition milk (the fifth postpartum milk sample). Individual milk serum samples [...] Read more.
The aim of this study was to evaluate the influence of applying a 4-week instead of an 8-week dry period to dairy cows on the proteome of colostrum (first sample) and of transition milk (the fifth postpartum milk sample). Individual milk serum samples of colostrum and transition milk were analysed from 12 Swedish Holstein (SH) and 12 Swedish Red (SR) cows. Samples were prepared by filter-aided sample preparation and dimethyl labelling and analysed by liquid chromatography tandem mass spectrometry. Shortening the dry period resulted in upregulation of 18 proteins in colostrum and transition milk of SR, whereas no statistical differences were found for SH colostrum and transition milk. These upregulated proteins may reflect a specific immune response in the SR samples that was reflected in colostrum as well as in transition milk. Upregulated proteins in colostrum seemed to reflect increased mammary epithelial cell proliferation in the periparturient period when a 4-week dry period was applied. The proteome data indicate that a dry period of 4 weeks to SR cows may not be sufficient for complete regeneration of the mammary epithelium. Full article
(This article belongs to the Section Dairy Systems Biology)
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