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Objective: To evaluate the complementary feeding practices, food intake, and nutritional status of infants on a cow’s milk protein elimination diet. Methods: A cross-sectional and observational study was conducted to compare infants aged 4-18 months who were on a cow’s milk protein elimination diet with a control group of healthy infants without any dietary restrictions. General information on the child’s health, demographic data, and food consumption were collected. Results: The study included 96 infants in the elimination diet group and 99 in the control group. In the elimination diet group, the median age (in months) of introduction of solid foods (5.0 × 4.0; p < 0.001) and water (5.5 × 4.0; p < 0.05) was later, consumption of soft drinks and industrialized cookies was less frequent (p < 0.05), and a lower index of complementary feeding inadequacies (2.75 × 3.50; p < 0.001) was observed. The elimination diet group presented lower individual values of Z scores for weight/age, weight/height, and body mass index/age, although they were fed with higher amounts of energy (117.4 × 81.3 kcal/kg of weight; p < 0.001) and macro-and micronutrients, except for vitamin A. In the elimination diet group, breast milk and its substitutes contributed to more than 67% of energy intake. Although calcium consumption was a deficit in 31.5% of the infants, none received supplementation. Conclusion: Infants on an elimination diet presented more adequate complementary feeding practices and higher nutritional intake, despite lower body weight values.

Introduction: This research was conducted to assess the effect of myo-inositol (MYO) in the freezing extender on the semen quality and oxidative stress parameters of frozen–thawed bull sperm. Mater...

Due to the diversity and limitation of determination methods, published data on the fatty acid (FA) compositions of different milk samples have contributed to inaccurate comparisons. In this study, we developed a high-throughput gas chromatography-mass spectrometry method to determinate milk FA, and the proposed method had satisfactory linearity, sensitivity, accuracy, and precision. We also analyzed the FA compositions of 237 milk samples from Holstein cows, Jersey cows, buffalos, yaks, humans, goats, donkeys, and camels. Holstein, Jersey, goat, and buffalo milks contained high content of even-chain saturated FA, whereas goat milk had higher content of medium- and short-chain FA (MSCFA). Yak and camel milk are potential functional foods due to their high levels of odd- and branched-chain FA and low ratios of n-6 to n-3 polyunsaturated FA (PUFA). Human milk contained lower levels of saturated FA, MSCFA, and conjugated linoleic acid, and higher levels of monounsaturated FA and PUFA. As a special nonruminant milk, donkey milk contained low levels of monounsaturated FA and high levels of PUFA and MSCFA. Based on the FA profiles of 8 types of milk, nonruminant milk was distinct from ruminant milk, whereas camel and yak milk were different from other ruminant milks and considered as potential functional foods for balanced human diet.

Even though supplementations of essential AA (EAA) are often related to increased lactose yields in dairy cows, underlying mechanisms connecting EAA availability to the mammary glands and lactose synthesis are poorly understood. The objective of this study was to examine the effects of branched-chain AA (BCAA) including Leu, Ile, and Val on (1) glucose transporter (GLUT1) abundance and glucose uptake, (2) the abundance of proteins regulating lactose synthesis pathway, and (3) fractional synthesis rates of lactose (FSR) using bovine mammary epithelial cells (BMEC) and mammary tissues slices (MTS). The BMEC (n = 4) were allocated randomly to regular Dulbecco's Modified Eagle Medium with Ham's F12 (DMEM/F12) medium (+EAA) or +EAA deficient (by 90%) in all EAA (- EAA), all BCAA (-BCAA), only Leu (-Leu), only Ile (-Ile) or only Val (-Val). Western immunoblotting analyses, depletion of glucose in media, and a proteomic analysis were performed to determine the abundance of GLUT1 in the cell membrane, net glucose uptake, and the abundance of enzymes involved in lactose synthesis pathway in BMEC, respectively. The MTS (n = 6) were allocated randomly to DMEM/F12 medium having all EAA and C-13-glucose at concentrations similar to plasma concentrations of cows (+EAA(p)), and +EAA(p) deprived of all BCAA (-BCAA(p)) or only Leu (-Leup) for 3 h. The C-13 enrichments of free glucose pool in MTS (EGlu-free) and the enrichments of glucose incorporated into lactose in MTS and media [ELactose-bound ((T&M))] were determined and used in calculating FSR. In BMEC, -BCAA increased the fraction of total GLUT1 translocated to the cell membrane and the fraction that was potentially glycosylated compared with +EAA. Among individual BCAA, only-Leu was associated with a 63% increase in GLUT1 translocated to the cell membrane and a 40% increase in glucose uptake of BMEC. The-BCAA tended to be related to a 75% increase in the abundance of hexokinase in BMEC. Deprivation of Leu tended to increase glucose uptake of MTS but did not affect EGlu-free, ELactose-bound ((T&M)), or FSR relative to +EAA(p). On the other hand, -BCAA(p) did not affect glucose uptake of MTS but was related to lower ELactose-bound (T&M), or FSR relative to +EAA(p). Considering together, decreasing Leu supply to mammary tissues enhances GLUT1 and thus glucose uptake, which, however, does not affect lactose synthesis rates. Moreover, the deficiency of other BCAA, Ile, and Val alone or together with the deficiency of Leu seemed to decrease lactose synthesis rates without affecting glucose uptake. The data also emphasize the importance of addressing the effect of the supply of other nutrients to the mammary glands than the precursor supply in describing the synthesis of a milk component.

Preconditioning of sperm using sub-lethal oxidative stress before cryopreservation is an innovative approach that can improve sperm cryo-survival. Mitochondrial uncoupling proteins (UCPs) are critical in reducing ROS level during stress conditions. The aim of the current study was to investigate whether mild sub-lethal stress induced by low concentrations of nitric oxide and hydrogen peroxide has a protective effect on quality parameters of post–thaw bull semen through modulations of mitochondrial uncoupling protein 2 (UCP2) expression. Semen samples were collected from 6 mature Holstein bulls, then mixed and divided into 8 aliquots: fresh, frozen control and frozen groups treated with NO: 0.1 (NO-0.1), 1(NO-1), 10 μ M (NO-10), and H2O2: 0.1(H2O2-0.1), 1(H2O2-1) and 10 μ M (H2O2-10). A significantly higher percentage of total motility, progressive motility and viability was observed in NO-1 and H2O2-10 compared to the other frozen groups (P μ M NO and 10 μ M H2O2 showed significantly increased percentages of mitochondria activity and membrane integrity (P

Carnosine is a naturally occurring dipeptide and a functional component in foods, while also showing health-promoting effects. Generally, food-derived carnosine is quantified via high-performance liquid chromatography (HPLC). We have developed a method for quantifying carnosine in foods using microbial enzymes, β-Ala-Xaa dipeptidase (BapA) and histidine ammonia-lyase (HAL). The carnosine concentrations in extracts of chicken, pork, beef, bonito, and tuna were determined via both HPLC and enzymatic determination. The carnosine contents measured via enzymatic determination were in agreement with those determined via conventional HPLC analysis. Relative standard-deviation values of the conventional HPLC method and the enzymatic determination of carnosine in foods were 0.728-5.76% and 0.504-4.58%, respectively. The recovery of carnosine in food extracts via enzymatic determination was 97-103%. Therefore, the developed enzymatic determination technique using BapA and HAL can be used for the determination of carnosine in meats and fishes with comparable accuracy to that of conventional HPLC analysis.

The objectives of the present study were (1) to evaluate the effect of prepartum cholecalciferol treatment on serum Ca concentration during the first 10 d after calving and (2) to evaluate the effect of treatment on subsequent health and performance. Multiparous Holstein cows (n = 377) from one dairy farm were fed a negative dietary cation-anion difference diet (−31 mEq/kg of DM) for the last 21 d of gestation. On d 275, the animals were randomly assigned to a control or a treatment group. Cows in the control group were left untreated, and cows in the treatment group received an injection of 12 × 106 IU of cholecalciferol intramuscularly on the day of enrollment. If treated cows did not deliver the calf within 6 d, they were reinjected with 10 × 106 IU of cholecalciferol. Blood samples were drawn on 1, 2, 3, 5, 7, and 10 days in milk (DIM) and analyzed for serum Ca, P, and Mg concentrations. In a subsample of cows (50 control cows, 35 cows treated once with cholecalciferol, and 15 cows treated twice) serum haptoglobin, nonesterified fatty acids, β-hydroxybutyrate, and 25-hydroxycholecalciferol concentrations were analyzed on 1, 5, and 10 DIM. Binary data [retained placenta (RP), metritis] were analyzed using logistic regression models. Repeated measures ANOVA with first-order autoregressive covariance was performed to evaluate the treatment effect on milk yield over the first 10 test days after parturition, 25-hydroxycholecalciferol, serum Ca, P, Mg, β-hydroxybutyrate, nonesterified fatty acids, and haptoglobin concentrations. Cox proportional hazards were used to model the time to event outcomes (time to pregnancy within 200 d, culling until 300 DIM). After enrollment of 31.4% of cows and a preliminary analysis, adverse reactions became apparent, and the study was stopped. Cows treated with cholecalciferol had a greater risk of incurring RP and metritis. The adjusted mean incidences were 2.0%, 7.7%, and 4.0% for RP, and 21.6%, 39.3%, and 33.3% for metritis for control cows, cows treated once, and cows treated twice with cholecalciferol, respectively. Compared with control cows, cows injected once with 12 × 106 IU of cholecalciferol produced less energy-corrected milk on the first (−3.76 kg) and second (−2.75 kg) test days, respectively. Cows injected twice with cholecalciferol (12 × 106 IU of cholecalciferol and 10 × 106 IU 1 wk later) had a reduced milk yield only at first test day (−3.80 kg). Treatment with cholecalciferol led to a significant increase in 25-hydroxycholecalciferol on d 1, 5, and 10 after calving. Serum Ca and P concentrations were significantly increased in cows treated with cholecalciferol, but serum Mg concentrations were significantly reduced. Haptoglobin concentrations were significantly increased on 5 DIM in cows injected once with 12 × 106 IU of cholecalciferol. Although we observed no effect of treatment on culling until 300 DIM, time to pregnancy was delayed by 34 d in cows injected once with 12 × 106 IU of cholecalciferol. In the present study, injection with 12 × 106 IU of cholecalciferol had detrimental effects on health and milk production despite the beneficial effects on Ca homeostasis.

Peripartum metabolism and subsequent reproductive performance of dairy cows are linked, with maladaptation over the transition period associated with poor reproductive success. A herd of seasonal calving, grazing dairy cows was established that differed in their genetic merit for fertility traits. The heifers were produced by a customized mating program to achieve a 10-percentage point divergence in the New Zealand fertility breeding value (FertBV) as follows: +5 FertBV (POS) versus −5% FertBV (NEG), while also limiting divergence in other breeding values, including body weight, body condition score, and milk production. In this study, we aimed to characterize differences in metabolic, mineral, and metabolic stress marker profiles during their first postpartum transition period as primiparous heifers and to examine if animals with longer postpartum anestrous intervals (PPAI; more than 66 d compared with less than 35 d) had greater metabolic dysfunction. Blood was sampled at −21, −14, −7, 0, 4, 7, 10, 14, 17, 21, 28, and 35 d relative to calving in 455 primiparous cows and plasma analyzed. The NEG cows had lower concentrations of both plasma nonesterified fatty acids and β-hydroxybutyrate at d 7 compared with POS cows. Detailed temporal profiling of various metabolic, mineral, and metabolic stress markers was undertaken in a subset of cows (n = 70). Cows were selected retrospectively to create 4 groups in a 2 × 2 factorial design with either a POS or NEG FertBV and either a short (19–35 d) or long (66–131 d) PPAI. The NEG cows tended, on average, to have lower nonesterified fatty acids and β-hydroxybutyrate concentrations compared with POS cows across the transition period. Mean body weight and body condition score was greatest in NEG cows when compared with the POS cows and an interaction with day demonstrated this only occurred precalving. They also had indications of improved liver health precalving, with higher albumin-to-globulin ratios and lower bilirubin concentrations. Concentrations of aspartate aminotransferase were lower, and the Na-to-Cl ratio was greater in cows with a long versus a short PPAI at d 28 and d 35 after calving, potentially because of cows with a short PPAI (19–35 d) returning to estrous during this time. Magnesium concentrations were lower in NEG cows with a short PPAI from d 21 onwards, indicating NEG cows may metabolically respond to estrous differently than POS cows. The NEG-long PPAI cows had greater gamma-glutamyl transferase concentrations from calving until d 28 and lower bilirubin concentrations throughout the transition period. Together, the results demonstrate significant effects of FertBV on peripartum metabolic status. However, most of the markers tested returned to reference intervals within 4 d after calving or remained within those intervals for the whole transition period, indicating relatively minor biological effects of FertBV on transition period adaptation. The profound differences in reproductive performance among the groups was not explained by underlying differences in metabolic responses during the transition period.

Nuclear magnetic resonance spectroscopy was applied to investigate the association between milk metabolome and udder quarter health status in dairy cows. Mammary gland health status was defined by combining information provided by traditional somatic cell count (SCC) and differential SCC (DSCC), which expresses the percentage of neutrophils and lymphocytes over total SCC. Quarter milk samples were collected in triplicate (d 1 to 3) from 10 Simmental cows, 5 defined as cases and 5 defined as controls according to SCC levels at d 0. A total of 120 samples were collected and analyzed for bacteriology, milk composition, SCC, DSCC, and milk metabolome. Bacteriological analysis revealed the presence of mostly coagulase-negative staphylococci in quarter milk samples of cows defined as cases. Nuclear magnetic resonance spectra of all quarter samples were first analyzed using the unsupervised multivariate approach principal component analysis, which revealed a specific metabolomic fingerprint of each cow. Then, the supervised cross-validated orthogonal projections to latent structures discriminant analysis unquestionably showed that each cow could be very well identified according to its milk metabolomic fingerprint (accuracy = 95.8%). The comparison of 12 different models, built on bucketed 1-dimensional NOESY spectra (noesygppr1d, Bruker BioSpin) using different SCC and DSCC thresholds, corroborated the assumption of improved udder health status classification ability by joining information provided by both SCC and DSCC. Univariate analysis performed on the 34 quantitated metabolites revealed lower levels of riboflavin, galactose, galactose-1-phosphate, dimethylsulfone, carnitine, hippurate, orotate, lecithin, succinate, glucose, and lactose, and greater levels of lactate, phenylalanine, choline, acetate, O-acetylcarnitine, 2-oxoglutarate, and valine, in milk samples with high somatic cells. In the 5 cases, results of the udder quarter with the highest SCC compared with its symmetrical relative were in line with quarter-level findings. Our study suggests that increased SCC is associated with changes in milk metabolite fingerprint and highlights the potential use of different metabolites as novel indicators of udder health status and milk quality.

We evaluated the effects of a constant low (5-5%) and modulated (5–20%) oxygen environments on the in vitro development of bovine oocyte-cumulus-granulosa cell complexes (OCGCs) cultured in the presence or absence of an antioxidant (astaxanthin: Ax). OCGCs were cultured in a gas permeable culture device for 8 days in 5-5% O2 (±Ax) and 5–20% O2 (±Ax) culture conditions. In the oxygen modulated culture conditions, the oxygen concentration was switched from 5% to 20% on day 4 of culture. Ax promoted the viability of OCGCs (P 0.05). A constant low oxygen environment significantly promoted the blastocyst rates (P 0.05). In conclusion, exposure of OCGCs to constant low oxygen or oxygen modulation in the presence of Ax promotes the healthy development of OCGCs during the 8-day IVG culture using the gas permeable culture device.

Flunixin is a nonsteroidal anti-inflammatory drug approved for use in cattle to manage pyrexia associated with bovine respiratory disease, mastitis, and endotoxemia. In the United States, no nonsteroidal anti-inflammatory drugs are approved for use in goats, but analgesics are needed for management of painful conditions to improve animal welfare. The objective of this study was to evaluate the pharmacokinetics of transdermal flunixin in dairy goats to determine a milk withdrawal interval (WDI) to avoid violative residue contamination in the food supply. Six adult lactating dairy goats received 3.3 mg/kg of transdermal flunixin before milk, interstitial fluid (ISF), and blood samples were collected at various time points for 360 h. The samples were analyzed using tandem mass spectrometry to detect flunixin as well as the flunixin marker metabolite, 5-hydroxyflunixin followed by a pharmacokinetic WDI calculation using the US Food and Drug Administration tolerance limit method to propose safe residue levels in goat milk. The mean flunixin apparent plasma half-life was 21.63 h. The apparent milk half-life for 5-hydroxyflunixin was 17.52 h. Our findings provide a milk WDI of 60 h using the US Food and Drug Administration tolerance of 0.002 µg/mL (established for bovine milk) and a more conservative WDI of 96 h using a limit of quantification of 0.001 µg/mL following the extralabel use of transdermal flunixin in dairy goats.

The objective of this study was to investigate the effects of a milk byproduct–based starter pellet, high in lactose, on feed intakes, growth, and rumen development when calves were fed either a high or low level of milk. This was done by using a 2 × 2 factorial design, with 120 Holstein heifer calves randomly assigned to 1 of 2 milk treatments and 1 of 2 feed treatments upon enrollment at birth. Calves were either assigned to a low (LM) or high (HM) level of milk replacer by an automated milk-feeding system for individually-housed calves. The LM treatment calves were offered 6 L/d (150 g/L; 1.2 L, 5×/d), whereas HM treatment calves were offered 15 L/d (150 g/L; 3 L, 5×/d) of milk replacer. Calves were also assigned to a milk byproduct- (whey) based starter feed treatment (WF), or a grain-based starter ration (95% concentrate starter pellet, 5% chopped wheat straw) feed treatment (GF). All calves were offered their treatment feed starting on d 5, with water offered ad libitum from d 0. The WF treatment calves were offered 150 g/d of the milk byproduct–based pellet until they consumed the entirety for 2 out of 3 consecutive days. At this point, WF treatment calves received 150 g of the milk byproduct–based pellet top dressed onto the grain-based ration to allow for ad libitum feed consumption until weaned, when they only received the grain-based ration postweaning (d 64–77). Calves on the GF feed treatment received only the grain-based ration. All calves were gradually weaned from d 43 to 63. By design, calves on the HM treatment consumed greater levels of milk than LM treatment calves for the majority of the days in the preweaning phase (8.9 ± 0.11 vs. 5.5 ± 0.11 L/d; SE). Calves on the GF and WF treatments consumed similar amounts of milk in the preweaning phase (7.2 ± 0.11 vs. 7.2 ± 0.11 L/d); however, there was a milk and feed treatment interaction detected for milk consumption in the weaning phase, with HM-GF calves drinking more milk than the other treatments and HM-WF calves drinking more milk than both LM-GF and LM-WF calves. The WF treatment calves had greater solid feed intake during the preweaning phase than GF treatment calves, with no feed intake differences in the weaning or postweaning phases. The WF calves drank more water in the preweaning phase than GF calves. Average daily gain was similar throughout the study between the GF and WF treatment calves. Rumen physiological measures were all similar by the end of the trial (d 77) regardless of feed or milk treatment, suggesting similar rumen development postweaning. Overall, regardless of the level of milk fed, offering the milk byproduct–based starter pellet may increase preweaning feed intake, along with water consumption. However, both feed and milk treatments had similar performance and rumen development measures by the end of the trial.

Metabolic disorders in early lactation have negative effects on dairy cow health and farm profitability. One method for monitoring the metabolic status of cows is metabolic profiling, which uses associations between the concentrations of several metabolites in serum and the presence of metabolic disorders. In this cross-sectional study, we investigated the use of mid-infrared (MIR) spectroscopy of milk for predicting the concentrations of these metabolites in serum. Between July and October 2017, serum samples were taken from 773 early-lactation Holstein Friesian cows located on 4 farms in the Gippsland region of southeastern Victoria, Australia, on the same day as milk recording. The concentrations in sera of β-hydroxybutyrate (BHB), fatty acids, urea, Ca, Mg, albumin, and globulins were measured by a commercial diagnostic laboratory. Optimal concentration ranges for each of the 7 metabolites were obtained from the literature. Animals were classified as being either affected or unaffected with metabolic disturbances based on these ranges. Milk samples were analyzed by MIR spectroscopy. The relationships between serum metabolite concentrations and MIR spectra were investigated using partial least squares regression. Partial least squares discriminant analyses (PLS-DA) were used to classify animals as being affected or not affected with metabolic disorders. Calibration equations were constructed using data from a randomly selected subset of cows (n = 579). Data from the remaining cows (n = 194) were used for validation. The coefficient of determination (R2) of serum BHB, fatty acids, and urea predictions were 0.48, 0.61, and 0.90, respectively. Predictions of Ca, Mg, albumin, and globulin concentrations were poor (0.06 ≤ R2 ≤ 0.17). The PLS-DA models could predict elevated fatty acid and urea concentrations with an accuracy of approximately 77 and 94%, respectively. A second independent validation data set was assembled in March 2018, comprising blood and milk samples taken from 105 autumn-calving cows of various breeds. The accuracies of BHB and fatty acid predictions were similar to those obtained using the first validation data set. The PLS-DA results were difficult to interpret due to the low prevalence of metabolic disorders in the data set. Our results demonstrate that MIR spectroscopy of milk shows promise for predicting the concentration of BHB, fatty acids, and urea in serum; however, more data are needed to improve prediction accuracies.

We hypothesised that heifers and cows with positive genetic merit for fertility would have a follicular microenvironment that resulted in better quality oocytes. To test this, we compared cumulus cell-oocyte complexes (COC) and follicular fluid from preovulatory follicles of 36 Holstein-Friesian nulliparous heifers and 50 primiparous lactating cows with either positive (POS, +5%) or negative (NEG, −5%) fertility breeding values (FertBV). Established gene markers of oocyte quality were measured in individual cumulus cell masses and oocytes, and concentrations of amino acids, steroids, and metabolites were quantified in corresponding follicular fluid and plasma. The timing of visually detectable oestrus in NEG FertBV heifers was inconsistent with their stage of COC maturation. Retrospective analyses of oestrous activity data indicated that NEG FertBV heifers were sampled earlier. Their recovered COC were morphologically less mature and exhibited differential expression of genes that are associated with follicular maturation (lower levels of BMPR2) and protein processing (higher levels of HSP90B1). Despite consistent sampling times being achieved in the lactating cows, lower concentrations of serine, proline, methionine, isoleucine, and non-esterified fatty acids were present in follicular fluid from POS FertBV cows. This was associated with higher expression of gene biomarkers of good oocyte quality (VCAN, PDE8A) in COC recovered from POS FertBV cows. This study supports our hypothesis that the follicular microenvironment in lactating dairy cows with high genetic merit leads to COC with higher metabolic rates and oocytes of superior quality. Moreover, an additional stressor such as lactation is required for this difference to be pronounced.

AIM A novel bone adhesive (tetracalcium phosphate and O-phospho-L-serine) has been developed as an osteoconductive, biodegradable bone-adherent material. The purpose of this study was to evaluate the maintenance of crestal bone/material level by standardized radiographs. METHODS This was a randomized, controlled, three arm, prospective study. Twenty-six mixed breed hound dogs were included in this study. Three implants were placed on either side of the mandible with either bone adhesive (BA), bovine bone mineral (BBM), or no biomaterial (NC). Standardized periapical radiographs were taken immediately after implant placement and at every month up to one year. The vertical distance between the implant platform to the first radiopaque material on both the mesial and distal surfaces were measured and crestal bone/material level changes were analyzed. RESULTS The crestal bone/material level adjacent to BA was stable and maintained throughout the study. There were statistically significant differences found between BA and NC in terms of maintenance of crestal bone levels at any given timepoint. CONCLUSION This study demonstrated that BA maintained crestal bone levels and had a similar ability to maintain that level over one year compared to BBM. This article is protected by copyright. All rights reserved.

The aim of the study was to investigate serum and milk concentrations of tryptophan (TRP), kynurenine (KYN) and kynurenic acid (KYNA), and activity of indoleamine 2,3-dioxygenase (IDO) in cows suffering from mastitis caused by Streptococcus spp. The blood and milk samples were collected from Holstein-Friesian cows farmed in the Lublin region of Poland. It was found that TRP was lower in cows with mastitis both in serum and milk. KYN was lower in serum but not in milk. KYNA was not significantly altered in diseased cows both in serum and milk. The activity of IDO calculated as KYN to TRP ratio was unchanged in serum but was markedly elevated in milk of cows with mastitis. Our findings may have important implications for diagnosis of mastitis in cows because an increase of activity of IDO and reduction of TRP in milk might be a valuable early marker predicting the occurrence of the disease.

Lipid droplets (LDs) consist of a core of neutral lipids such as triacylglycerols and cholesteryl esters covered by a phospholipid monolayer. Recent studies have shown that LDs not only store neutral lipids but are also associated with various physiological functions. LDs are found in most eukaryotic cells and vary in size and quantity. It has long been known that mammalian oocytes contain LDs. Porcine and bovine oocytes contain substantial amounts of LDs, which cause their cytoplasm to darken, whereas mouse and human oocytes are translucent due to their low LD content. A sufficient amount of LDs in mammalian oocytes has been thought to be associated with oocyte maturation and early embryonic development, but the necessity of LDs has been questioned because embryonic development proceeds normally even when LDs are removed. However, recent studies have revealed that LDs play a crucial role during implantation and that maintaining an appropriate amount of LDs is important for early embryonic development, even in mammalian species with low amounts of LDs in their oocytes. This suggests that a fine-tuned balance of LD content is essential for successful mammalian embryonic development. In this review, we discuss the physiological importance of LDs in mammalian oocytes and preimplantation embryos based on recent findings on LD biology.

Building compatible surface on perovskite quantum dots (PQDs) for applications like sensing analytes in aqueous medium is highly challenging and if achieved by simple means can revolutionize disease diagnostics. The present work reports the surface engineering of CsPbBr3 QDs via "simple ligand exchange process" to achieve water-compatible QDs towards detection of biomolecules. The monodentate oleic acid ligand in CsPbBr3 QDs is exchanged with dicarboxylic acid containing (bidentate) ligands such as folic acid (FA), ethylenediamine tetra-acetic acid (EDTA), succinic acid (SA) and glutamic acid (GA) to develop an efficient water-compatible PQD-ligand system. optical and theoretical studies showed the existence of a stronger binding between the perovskite and succinic acid ligand as compared to oleic acid (OA) and all other ligands. Replacement of OA with SA and retention of crystal structure is validated using spectroscopic and microscopic tools. It is observed that SA ligands facilitate better electronic coupling with PQDs and show significant improvement in fluorescence and stability. Further N-Hydroxy succinimide (NHS), which is a well-known compound to activate carboxyl groups, is used to bind onto SA PQDs as multidentate ligand, to form water stable PQDs. SA PQDs react with NHS (in water) to form multidentate ligand passivated PQDs that show very high photoluminescence (PL) as compared to OA PQDs in toluene. This also results in the formation of an NHS ester that allows bioconjugation with PQDs. This simple probe in water is further utilized for sensing a highly hydrophilic bovine serum albumin (BSA) protein as a model target to demonstrate the potential and effectiveness of this process to create compatible QDs for the successful conjugation of biomolecules. Although the focus of this work is to demonstrate bioconjugation and not achieving higher sensitivity levels, the intrinsic sensing level of these compatible QDs towards BSA shows a detection limit of 51.47 nM, which is above par with other reports in literature.

Many male dairy calves are castrated when reared for beef production, but for dairy breeds the assessment of the longer-lasting pain associated with this procedure has received little scientific attention. In this study we assessed 2 methods: surgical (n = 10 calves) and rubber ring (n = 11). All calves were castrated at 28 d of age using multimodal pain control. During the 8 wk that followed, we recorded wound healing, local inflammation, body weight, milk and calf starter intake, lying time, and wound-directed behavior. Surgical wounds were fully healed on average 4 wk after the procedure, but only 1 calf in the rubber ring treatment fully healed within the 8-wk study. Inflammation was greater after rubber ring castration; skin temperature in the area around the lesion was 1.7 ± 0.35°C (mean ± standard deviation) higher than for the surgical treatment. Compared with surgically castrated calves, those castrated by rubber ring gained less weight over the study period (on average 11.9 ± 5.1 kg less), a difference due in part to lower intake of calf starter (on average 1.8 ± 0.6 kg less). Calves in the rubber ring treatment spent less time lying down (on average 4.2 ± 1.2% fewer scans per day) and licked their lesions more frequently (on average 16.0 ± 3.3 more licks per day). We conclude that the rubber ring calves experienced more pain in the weeks following the procedure and thus recommend that surgical castration be favored for preweaning dairy calves.

Background: This study aimed to evaluate the effect of the combination of zinc oxide nanoparticles (NPs) and chitosan NPs on the shear bond strength (SBS) of composites used for orthodontic bonding. Methods: Four groups of composites (n = 10), containing 0%, 1%, 5%, and 10% w/w NP fillers, respectively, were used to bond brackets to the surfaces of 40 intact bovine incisors. After 1000 rounds of thermal cycling at 5°C–55°C, all specimens were mounted in acrylic blocks. The SBS was tested using a universal testing machine, and the adhesive remnant index scores were registered using a stereomicroscope. Data were statistically analyzed using a 1-way ANOVA and the Kruskal–Wallis test. Results: The highest value of mean SBS was found in the control group, and the lowest value was found in the group with composite containing 10% NPs. The adhesive remnant index did not differ significantly among the groups (P = 0.823). Conclusions: Incorporation of 1% and 5% zinc oxide and chitosan NPs had no effect on the SBS of composite, and the obtained SBS values were similar to that of the control group.

Emerging concern about the emergence of antimicrobial resistance has limited the use of antibiotics in calves. Hence, there is a need to find suitable alternatives to antibiotics to manage gastrointestinal infections in neonatal calves. The objective of the present study was to develop a probiotic of calf-origin for its potential application in calf nutrition. Accordingly, 69 lactic acid bacteria (LAB) strains were isolated from faeces of newborn calves, out of which 10 strains were short-listed for further in vitro testing based on the aggregation time and cell surface hydrophobicity. The results of acid-, bile- and phenol-tolerance tests indicated that out of the ten strains, the isolate CPN60 had better resistance to these adverse conditions likely to be encountered in the gastrointestinal tract. The isolate also showed an optimal ability to produce biofilm. Further assessments reiterated its superiority in terms of co-aggregation and antagonistic activity against pathogenic strains of Escherichia coli. Subsequently, the isolate was identified through 16S rRNA sequencing and sequence homology and designated as Ligilactobacillus salivarius CPN60. The candidate probiotic was evaluated in vivo using 48 male (5 weeks old) Wistar rats, divided into two equal groups viz. control (CON) and probiotic (PRO). During the 4-weeks feeding trial, the PRO group rats were gavaged with one mL culture of L. salivarius CPN60 equivalent to 108 CFU/rat. The in vivo trial results indicated better nutrient utilization efficiency and growth performance (p

The global burden of diarrhea caused by Cryptosporidium parasite is underestimated. In immunocompromised hosts, chronic and severe presentation of intestinal cryptosporidiosis can result in long-term morbidity and high illness costs. The evidence of effective treatments for cryptosporidiosis has been lacking. We reviewed the published clinical trials to bring forward the feasible therapeutic options of human cryptosporidiosis in various populations and settings according to clinical improvement and parasite clearance rates. A total of 42 studies consisting of the use of nitazoxanide, paromomycin, macrolides, somatostatin analogues, letrazuril, albendazole, rifaximin, miltefosine, clofazimine, and colostrum were included in the review. The trials were mostly conducted in small number of individuals infected with human immunodeficiency virus (HIV), and there is inadequate data of controlled trials to suggest the use of these treatment modalities. Nitazoxanide was reported to be highly efficacious only in immunocompetent hosts and was found to be superior to paromomycin in the same group of patients. Macrolides showed no effective results in both clinical and parasitological improvement. Human bovine colostrum should possibly be administered as one of complementary therapeutic modalities along with other antimicrobials to reach optimal parasite eradication. Other trials of therapeutic modalities were terminated due to futility. Currently, available data is intended to aid the development of strategies for improving access to treatments in different clinical settings, as well as to help guide further studies on treatments of human intestinal cryptosporidiosis., Graphical abstract Image 1

Our objective was to determine whether abomasal infusions of increasing doses of oleic acid (cis-9 C18:1; OA) improved fatty acid (FA) digestibility and milk production of lactating dairy cows. Eight rumen-cannulated multiparous Holstein cows (138 d in milk ± 52) were randomly assigned to treatment sequence in a replicated 4 × 4 Latin square design with 18-d periods consisting of 7 d of washout and 11 d of infusion. Production and digestibility data were collected during the last 4 d of each infusion period. Treatments were 0, 20, 40, or 60 g/d of OA. We dissolved OA in ethanol before infusions. The infusate solution was divided into 4 equal infusions per day, occurring every 6 h, delivering the daily cis-9 C18:1 for each treatment. Animals received the same diet throughout the study, which contained (percent diet dry matter) 28% neutral detergent fiber, 17% crude protein, 27% starch, and 3.3% FA (including 1.8% FA from a saturated FA supplement containing 32% C16:0 and 52% C18:0). Infusion of OA did not affect intake or digestibility of dry matter and neutral detergent fiber. Increasing OA from 0 to 60 g/d linearly increased the digestibility of total FA (8.40 percentage units), 16-carbon FA (8.30 percentage units), and 18-carbon FA (8.60 percentage units). Therefore, increasing OA linearly increased absorbed total FA (162 g/d), 16-carbon FA (26.0 g/d), and 18-carbon FA (127 g/d). Increasing OA linearly increased milk yield (4.30 kg/d), milk fat yield (0.10 kg/d), milk lactose yield (0.22 kg/d), 3.5% fat-corrected milk (3.90 kg/d), and energy-corrected milk (3.70 kg/d) and tended to increase milk protein yield. Increasing OA did not affect the yield of mixed milk FA but increased yield of preformed milk FA (65.0 g/d) and tended to increase the yield of de novo milk FA. Increasing OA quadratically increased plasma insulin concentration with an increase of 0.18 μg/L at 40 g/d OA, and linearly increased the content of cis-9 C18:1 in plasma triglycerides by 2.82 g/100 g. In conclusion, OA infusion increased FA digestibility and absorption, milk fat yield, and circulating insulin without negatively affecting dry matter intake. In our short-term infusion study, most of the digestion and production measurements responded linearly, indicating that 60 g/d OA was the best dose. Because a quadratic response was not observed, improvements in FA digestibility and production might continue with higher doses of OA, which deserves further attention.

Cow milk allergy is one of the most prevalent food allergies worldwide, particularly in infants and children. To the best of our knowledge, minimal research exists concerning the antigenicity of cow milk (CM). This study was performed to evaluate the allergenicity of enzymatically hydrolyzed cow milk (HM) in a BALB/c mouse model. The mice were randomly divided into 5 groups (n = 12/group), which were sensitized with phosphate-buffered saline, CM, and HM (Alcalase-, or Protamex-, or Flavorzyme-treated cow milk; Novo Nordisk; AT, PT, FT, respectively), respectively, using cholera toxin as adjuvant on d 0, 7, 14, 21. On d 28, the test mice were orally challenged with phosphate-buffered saline, CM, and HM (AT, PT, or FT) alone. Anaphylactic symptoms were monitored in the mice. Antibody, cytokine, histamine, and mouse mast cell protease-1 (mMCP-1) levels were measured using enzyme-linked immunosorbent assays. In addition, the numbers of T helper (Th)1 and Th2 cells, as well as the proportions of CD4+CD25+Foxp3+Tregs cells, in mouse spleens were detected using flow cytometry. Statistical significance was determined by one-way ANOVA. The results revealed significant differences between CM- and HM-challenged mice. Among these, the clinical scores of HM-challenged mice (AT, 1.50; PT, 2.00; FT, 1.92) were lower than those of CM-challenged mice (positive control, 2.83), but body weight and temperature of HM-challenged mice were higher than those of CM-challenged mice. In addition, significant reductions of allergen-specific IgE, IgG, histamine, and mMCP-1 were showed in HM-challenged mice, especially for histamine, ranging from 171.42 ng/mL to 214.94 ng/mL. Remarkable reductions of IL-4, IL-5, and IL-13 levels, as well as elevations of interferon-γ and IL-10 levels in the spleens of HM-challenged mice were also detected. Moreover, the number of Th2 cells decreased in the HM-challenged mice, to 2.36% (AT), 1.79% (PT), and 4.03% (FT), respectively, whereas the numbers of Th1 cells (AT, 6.30%; PT, 6.70%; FT, 6.56%) and the proportions of CD4+CD25+Foxp3+Tregs (AT, 8.86%; PT, 9.21%; FT, 9.16%) increased significantly. Our findings indicate that exposure to HM was sufficient to induce a shift toward a Th1 response, thereby reducing potential allergenicity. Importantly, these results will lay a theoretical foundation for the development of hypoallergenic CM products.

Our objective was to evaluate the effect of metabolizable protein (MP) supply on milk production, blood metabolites, and health in dairy cows during early lactation. Three experimental diets were formulated to contain 114, 107, 101 g of MP/kg of dry matter (DM; 114MP, 107MP, and 101MP, respectively) with crude protein contents of 17.0, 16.2, and 15.3% of DM, respectively. One hundred multiparous Holstein cows were fed 1 of these 3 diets during wk 1 to 3 and wk 4 to 13 of lactation in one of the following sequences: (1) 114MP and 107MP (114MP/107MP), (2) 114MP and 101MP (114MP/101MP), or (3) 101MP and 101MP (101MP/101MP). During wk 1 to 3, the 114MP and 101MP treatments were 20 and 27% deficient in estimated MP, respectively. From wk 4 to 13, the 114MP/107MP, 114MP/101MP, and 101MP/101MP treatments were 8, 12, and 13% deficient in estimated MP, respectively. Data were analyzed separately for wk 1 to 3, 4 to 13, and 1 to 13. Dry matter intake and energy-corrected milk (ECM) yield were not affected by treatment during wk 4 to 13 or wk 1 to 13; however, ECM yield decreased for 101MP versus 114MP from wk 1 to 3. Similarly, feed efficiency was not affected by treatment from wk 4 to 13 or wk 1 to 13, and was reduced with 101MP versus 114MP during wk 1 to 3. Milk N efficiency tended to increase for 101MP versus 114MP for wk 1 to 3 and increased with 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13. Treatment had no influence on yields and concentrations of milk components from wk 4 to 13 or wk 1 to 13; however, compared with 114MP, feeding 101MP tended to decrease milk fat yield and decreased yields of milk true protein and lactose for wk 1 to 3. Both milk and blood urea N concentrations decreased for 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13, and were reduced with feeding 101MP versus 114MP from wk 1 to 3. Treatment had no effect on the incidence of diseases in cows throughout the study. Serum concentrations of total fatty acids, albumin, and aspartate aminotransferase did not differ between 101MP and 114MP; however, serum β-hydroxybutyrate concentration was lower in cows receiving 101MP during the first 3 wk of lactation. Compared with 114MP, feeding 101MP during wk 1 to 3 increased plasma concentrations of creatinine and 3-methylhistidine (3-MHis) but did not change the ratio of plasma 3-MHis to creatinine. We found no differences in plasma creatinine or the ratio of 3-MHis-to-creatinine among treatments from wk 4 to 13; however, 101MP/101MP and 114MP/101MP had elevated plasma 3-MHis compared with 114MP/107MP. Treatment had no effect on body weight and body condition score over the duration of the study. Collectively, despite reduced milk production for the first 3 wk of lactation, feeding the 101MP/101MP treatment sustained lactational performance and improved milk N efficiency without negatively affecting the frequency of diseases in dairy cows during the first 13 wk postpartum.

Granulosa cells (GCs) are essential somatic cells in the ovaries, and apoptosis of GCs causes follicular atresia. microRNA-10b (miR-10b) is pivotal for cell apoptosis. However, currently, little is known about the role of miR-10b in bovine ovarian GCs (BGCs). In this study, the effect of miR-10b on the apoptosis of BGCs was investigated. Our results showed that the overexpression of miR-10b could increase the apoptosis rate of BGCs, which is associated with the increased expression of Caspase-3 and decreased expression ratio of Bcl-2/Bax (P 0.05). Furthermore, plasminogen activator inhibitor-1 (PAI-1) was confirmed to be a validated target of miR-10b in BGCs using dual-luciferase reporter analysis, and transfection of miR-10b mimics decreased the expression of PAI-1 (P 0.05). In addition, overexpression of PAI-1 significantly inhibited BGC apoptosis (P 0.05), and PAI-1 could alleviate BGC apoptosis induced by miR-10b (P 0.05). Subsequently, phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) was found to be the downstream pathway of PAI-1 by RNA-Seq analysis and verified by Western blot. Finally, a PI3K/AKT inhibitor (Miltefosine) was used to inhibit the PI3K/AKT pathway, which reversed the inhibitory effect of PAI-1 on the apoptosis of BGCs (P 0.05), and enhanced the promotion effect of miR-10b on the apoptosis of BGCs (P 0.05). Our results indicated that miR-10b promotes BGC apoptosis by targeting PAI-1 to regulate the PI3K/AKT pathway.

We aimed to evaluate the effects of feeding super-conditioned corn at different temperatures on intake, growth performance, total-tract starch digestibility, rumen fermentation, blood metabolites, and feeding behavior of dairy calves. Thirty-six Holstein female dairy calves (40 ± 1.72 kg of body weight, ± SD) were randomly assigned to 1 of the following 3 treatments: (1) ground corn (control; CON; n = 12), (2) corn super-conditioned at 75°C (T-75; n = 12), and (3) corn super-conditioned at 95°C (T-95; n = 12). Three mash starter feeds with an identical nutritional composition were blended with 5% chopped alfalfa hay and fed to individually-housed calves from d 3 to 77 of their birth. All calves were fed 4 L/d of pasteurized whole milk twice daily since d 3 to 56, followed by 2 L/d of morning feeding from d 57 to 63 of age. Calves were weaned on d 63 and remained in the study until d 77. The T-75 and T-95 diets increased total-tract starch digestibility compared with the CON diet. Dry matter intake and weaning or final BW were not affected by treatments; however, average daily gain and feed efficiency increased in calves fed T-95 in the overall period. The T-95 diet increased withers height and tended to increase hip height compared with other diets, but feeding behavior did not change throughout the experimental period. Ruminal pH decreased in calves fed the T-95 diet compared with T-75 and CON diets. The molar proportion of ruminal propionate increased, whereas the acetate-to-propionate ratio tended to decrease in calves fed the T-95 compared with CON diet. Calves fed the T-95 diet had the highest blood glucose concentration, whereas a trend for increased insulin concentration was observed in calves fed T-95 compared with other diets. In conclusion, super-conditioning temperature of corn (T-95 vs. T-75 and CON) improved the average daily gain, feed efficiency, and skeletal growth, but did not influence dry matter intake during the first 77 d of age. Finally, the total-tract starch digestibility increased, whereas ruminal pH dropped during the postweaning period as super-conditioning temperature elevated.

Zilpaterol and clenbuterol are two β-adrenergic agonist drugs used in animal production. Both drugs have anabolic effects with advantages on carcass yield. Meanwhile, zilpaterol is approved for animal feed in authorized countries. Clenbuterol is a banned substance due to the risk of toxicity; however, it is still being used in unknown dose levels in many farm species. Therefore, the use and abuse of these substances should be closely monitored, considering the clenbuterol ability and the not proved yet of zilpaterol to produce reactive oxygen and nitrogen species. Regarding glutathione which is the main intracellular antioxidant plays detoxification functions on liver metabolism; in this work, it is our interest to know the capacity of chitosan-glutathione nanoparticles (CS/GSH-NP) as a complementary source of exogenous GSH to modify the oxide-reduction status on bovine precision-cut liver slice cultures (PCLS) exposed to clenbuterol and zilpaterol. A single drug assay was performed in first instance by adding clenbuterol, zilpaterol, chitosan nanoparticles (CS-NP), and CS/GSH-NP. Then combinate drug assay was carried out by testing clenbuterol and zilpaterol combined with CS-NP or CS/GSH-NP. The results showed that both β-adrenergic agonists modify in a dose-dependent manner in oxide-reduction response through ROS generation. The activity or content of glutathione peroxidase activity, intracellular GSH, gamma glutamyl-transpeptidase, aspartate aminotrasnferase and alanine aminotrasnferase were modified. The exogenous GSH delivered by nanoparticles could be used to modulate these markers.

Biosolids are a by-product of wastewater treatment, and their nutritional composition makes them ideal for fertilizing crops. However, pre-treatments, such as conditioning and/or (vermi)composting, are often required to stabilize the product and remove pathogens. Biosolids, cow manure, and a 50–50% mixture were conditioned for 21 days, composted or vermicomposted with Eisenia fetida (Savigny 1826) for 28 days, and applied to soil cultivated with wheat (Triticum sp. L.), while emissions of nitrous oxide (N2O), methane (CH4), and carbon dioxide (CO2) were monitored. Emissions of CH4 were large from the biosolid and N2O from the cow manure during conditioning. Emissions of CH4 remained high during (vermi)composting of the biosolids, while the emissions of N2O from the cow manure dropped. The addition of E. fetida did not affect the emissions of greenhouse gases during (vermi)composting. The emission of N2O was higher when (vermi)composted biosolid was applied to soil cultivated with wheat than when (vermi)composted cow manure was applied. The global warming potential (GWP) of the sum of the emitted greenhouse gases (GHG) during conditioning, (vermi)composting, and when the final product was applied to soil was 3 times larger from the cow manure than from the biosolid, but mixing biosolid with cow manure eliminated that difference. It was concluded that mixing biosolid with cow manure might be a simple way to reduce the GWP of the emitted GHG during storage, (vermi)composting, and when applied to soil.

The COVID-19 pandemic, which causes severe respiratory tract infections in humans, has become a global health concern and is spreading rapidly. At present, the most important issue associated with COVID-19 is the immune system and the factors that affect it. It is well known that cow’s milk is highly rich in micronutrients that increase and strengthen the immune system. Research shows that the administration of these nutrients is very effective in fighting COVID-19, and a deficiency in any of them can be a weakness in the fight against the virus. On the other hand, cow’s milk is accessible to the whole population, and drinking colostrum, raw, and micro-filtered milk from cows vaccinated against SARS-CoV-2 could provide individuals with short-term protection against the SARS-CoV-2 infection until vaccines become commercially available. This review aimed to discuss the effects of milk vitamins, minerals, and bioactive peptides on general health in humans to combat viral diseases, especially COVID-19, and to what extent cow’s milk consumption plays a role in providing these metabolites. Cow’s milk contains many bioactive compounds that include vitamins, minerals, biogenic amines, nucleotides, oligosaccharides, organic acids, and immunoglobulins. Humans can meet a significant portion of their requirements for vitamins and minerals through the consumption of cow’s milk. Recent studies have shown that micronutrients such as vitamins D, E, B, C, and A as well as minerals Zn, Cu, Mg, I, and Se and bioactive peptides, each can have positive and significant effects on strengthening the immune system and general health in humans.

BACKGROUND Faba bean varieties with low or zero tannin content have been developed in Canada to overcome the negative effects of condensed tannins on the utilization by ruminants of crude protein (CP) and starch. However, their nutritional value has not been evaluated for incorporation in dairy rations. The objectives of this study were to investigate (i) the chemical profile; (ii) the Cornell Net Carbohydrate and Protein System (CNCPS) protein and carbohydrate subfractions; (iii) the energy values; (iv) the ruminal, intestinal, and total digestibility of CP; (v) the metabolizable protein (MP) supply to dairy cows; and (vi) the protein-inherent molecular spectral characteristics of brown-seeded (var. Fatima) faba beans with high tannin content and yellow-seeded (var. Snowbird) faba beans with low tannin content. RESULTS The results revealed that Fatima beans had higher (P

Vigorous or prolonged exercise poses a challenge to gastrointestinal system functioning and is associated with digestive symptoms. This narrative review addresses 1) the potential of dietary supplements to enhance gut function and reduce exercise-associated gastrointestinal symptoms and 2) strategies for reducing gastrointestinal-related side effects resulting from popular sports supplements. Several supplements, including probiotics, glutamine, and bovine colostrum, have been shown to reduce markers of gastrointestinal damage and permeability with exercise. Yet the clinical ramifications of these findings are uncertain, as improvements in symptoms have not been consistently observed. Among these supplements, probiotics modestly reduced exercise-associated gastrointestinal symptoms in a few studies, suggesting they are the most evidenced-based choice for athletes looking to manage such symptoms through supplementation. Carbohydrate, caffeine, and sodium bicarbonate are evidence-based supplements that can trigger gastrointestinal symptoms. Using glucose-fructose mixtures is beneficial when carbohydrate ingestion is high (>50 g/h) during exercise, and undertaking multiple gut training sessions prior to competition may also be helpful. Approaches for preventing caffeine-induced gastrointestinal disturbances include using low-to-moderate doses (

Although milk and plant-based drinks are widely consumed foodstuffs with high nutritional value, their consumption may also mean intake of non-essential/toxic elements becoming a risk for human health. This study was aimed at determining the concentrations of essential (Ca, Co, K, Mg, Mn, Na, Ni and P) and non-essential/toxic (Hg, Pb, U and V) elements in milks (cow and goat), plant-based drinks (soy, almond, rice and oat) and infant formulas from organic and conventional production systems. Lactose-free, fresh and ultra-high-temperature (UHT) milks were also included. Chemical analyses were performed by means of inductively coupled plasma-mass spectrometry (ICP-MS). The content of the elements hereby assessed did not depend on the production system and the presence of lactose. However, significant differences were found in the concentrations of multiple elements when comparing sterilization methods, source (animal vs. plant-based) and animal species. Non-essential elements were not detected in milks and plant-based drinks, excepting Pb, which was detected in three samples. While the consumption of goat milk is recommended, considering the global intake of essential elements and the absence of non-essential elements, further studies should be conducted to confirm the absence of non-target toxic elements at very low trace levels. On the other hand, the best plant-based drinks are those made up with almonds (intake of Ca) and soy (K and Mg). The current results should be useful to help the population to balance the benefits and risks from milks and plant-based drinks consumption, as well as to adapt their dietary habits.

Neonatal calf mortality is a major concern to livestock sector worldwide. Neonatal calf diarrhoea (NCD), an acute severe condition causes morbidity and mortality in calves. Amongst various pathogens involved in NCD, E. coli is considered as one of the major causes. The study was targeted to characterize E. coli isolates from neonatal calves for diarrhoeagenic Escherichia coli (DEC) types (pathotyping), antimicrobial resistance (AMR) profiling and to correlate with epidemiological parameters. From neonates, a total of 113 faecal samples were collected, out of that 308, lactose fermenting colonies were confirmed as E. coli. Pathotypable isolates (12.3%) were represented by STEC (6.1%), EPEC (2.9%), ETEC (1.9%), EAEC (0.9%) and EHEC (0.3%). Occurrence of STEC was more in non-diarrhoeic calves, whereas ETEC was observed more in diarrhoeic calves. EPEC occurrence was observed in both diarrhoeic and non-diarrhoeic calves. Fishers extract test showed no significant association for occurrence of DEC types to type of dairies, health status, species, breed, age and sex of neonatal calves. Two hundred and eighty isolates were tested for antimicrobial susceptibility. The isolates showed maximum resistance towards ampicillin (55.4%) followed by tetracycline (54.3%), while minimum resistance was observed towards meropenem (2.5%). Multidrug resistant E. coli isolates were found to be 139 (49.6%), and Extended-spectrum beta-lactamase (ESBL) producers were 120 (42.9%). DEC pathotypes like STEC, ETEC, EHEC and EAEC that are also multidrug resistant present in neonatal calves have zoonotic potential and hence are of public health significance. Supplementary Information The online version contains supplementary material available at 10.1007/s11259-021-09857-5.

Studies have revealed that a novel anti-inflammatory mediator─maresin-1 (MaR1)─can reduce the level of inflammatory factors. There is evidence that physical exercise (PE) promotes the biosynthesis of MaR1, leading to the prevention of rheumatoid arthritis (RA). Previously, we have proven that resveratrol can mitigate the formation of RA. Pterostilbene (Pte) is an analogue of resveratrol, but it is around four times more bioavailable. Hence, we hypothesize that Pte could be more effective in preventing RA, in particular, when accompanied by moderate PE. Based on this hypothesis, we explored the preventive effect of Pte combined with PE on a bovine type II collagen (BIIC)-stimulated rat RA model and its underlying molecular mechanism. Compared with the BIIC-stimulated group, the serum content of MaR1 with continuous intervention of Pte plus PE for 8 weeks was significantly increased to 46.3 pg/mL from 7.2 pg/mL in BIIC-treated alone. Besides, the variation in the relative expression levels of p-NF-κB and p-Akt was reversed with the administration of Pte plus PE. More importantly, the in vitro results confirmed that the treatment of Pte plus MaR1 inhibited proliferation and apoptosis and promoted the autophagy of the interleukin (IL)-1β-stimulated primary rat synovial cells through the PI3K/Akt/NF-κB signal pathway. Collectively, the oral administration of Pte plus moderate PE helped to ameliorate the pathological process of RA by correcting the PI3K/Akt/NF-κB signal pathway.

AIM AND OBJECTIVE This study aimed to evaluate the effect of ozonated water used as an irrigant and dentin-cleaning solution on the bond strength of fiberglass pins in vitro, comparing it with the commonly used solutions. MATERIAL AND METHODS Seventy-seven bovine roots were randomly divided into seven groups according to the irrigant and dentin-cleaning solution to be used: HP/HP, 2.5% sodium hypochlorite; DA/DA, distilled water; CHX/CHX, 2% chlorhexidine; OA/OA, 4 ppm ozonated water; HP/DA; HP/CHX; and HP/OA. In each group, the root canals were endodontically prepared by using the corresponding irrigant and stored in DA; after 7 days, they were cleared and cleaned with the corresponding cleaning solution. The fiberglass pins were cemented by using self-adhesive cement. After 7 days, the roots were sectioned (six discs each) and submitted to the push-out test. The type of fracture was analyzed with a 4.5x stereoscopic magnifying glass. The data were analyzed by the analysis of variance and the Tukey test at a 5% significance level. RESULTS The OA/OA (11.67), HP/HP (11.21), and HP/OA (9.71) groups showed the highest mean push-out bond strength (MPa) in the cervical third. The same trend was maintained in the middle and apical thirds. CONCLUSION Ozonated water and 2.5% sodium hypochlorite are the most relevant solutions for root dentin treatment. CLINICAL RELEVANCE Teeth treated with ozonated water, sodium hypochlorite, and a combination of the two showed greater bond strength than those treated with other solutions.

Here, we established dedifferentiated fat (DFAT) cells from mature bovine adipocytes and then examined the effects of volatile fatty acids on the differentiation of these DFAT cells into adipocytes in vitro. When mature adipocytes were isolated from bovine adipose tissue and cultured using the ceiling culture method, they were dedifferentiated into fibroblast-like cells without lipid droplets. These fibroblast-like cells, termed bovine DFAT (b-DFAT) cells, actively proliferated. After adipogenic induction, increased expression of adipocyte-specific genes occurred in b-DFAT cells and they redifferentiated into adipocytes with an accumulation of lipid droplets in their cytoplasm. The effects of volatile fatty acids on adipocyte differentiation in b-DFAT cells were also examined. Specifically, acetate, butyrate, and propionate added to adipogenic induction medium significantly enhanced the adipogenesis of b-DFAT cells compared with that observed in control cells; the addition of 10-3 mol of acetate enhanced adipogenesis of b-DFAT cells to the greatest extent. These results suggest that b-DFAT cells derived from bovine mature adipocytes are appropriate for the study of bovine adipocyte differentiation and that the optimum concentration treatment of acetate, a major energy source for ruminants, promotes adipogenesis of b-DFAT cells in vitro.

Soy-cow blended milk is a potential nutritional beverage and raw material for dairy products. This study determined the particle size, flow, lubrication, flavour and sensory properties of cow milk, soy milk and their blends. Twenty-one major volatile compounds were identified using solid-phase microextraction gas chromatography (SPME-GCMS) in cow milk and soy milk. Among all the compounds detected in the milk samples, hexanal, associated with off flavour was found highest in soymilk followed by cow milk and blended milk. From confocal images, soy-cow blended milk at a ratio of 1:1 showed a homogenous distribution of small fat globules and protein compared to the soy milk and cow milk. The addition of soy milk to cow milk lowers the particle size although not significantly (p > 0.05) and decreases the viscosity of blended milk. Cow milk was the most viscous (2.66 mPa.s at 50 s ) with large particles (0.48 μm) observed from confocal images. However, soymilk was found to have better lubrication properties (boundary regime) with a lower friction coefficient (~0.30) compared to cow milk (~0.40) and blended milk (~0.50) at low entrainment speed (0.1-2 mm/s). The sensory panel ranked cow milk as creamier and more viscous while soymilk was perceived as more astringent with beany flavour. Overall, a proportion of 3:7 soy-cow blended milk was more acceptable than the other two blended milks with less beany flavour, as confirmed by the lower amount of hexanal from gas chromatography mass spectrometer. This article is protected by copyright. All rights reserved.

Bovine vaccinia (BV) is an infectious disease caused by Vaccinia virus (VACV) characterized by vesicular and exanthematic lesions, mainly in cattle. Although BV has been described in some Brazilian regions in the last decades, official information regarding the current prevalence in bovine herds of Midwestern Brazil is lacking. Thus, the current study aimed to estimate the seroprevalence and risk factors associated with BV in cattle in the Distrito Federal (DF), Brazil. Sera of 312 cows of 64 herds were tested by virus-neutralizing test for VACV antibodies. Herd and animal seroprevalence were estimated to be 33.3% (CI 95%: 18.2–48.3%) and 10.6% (CI 95%: 1.0–20.2%), respectively. Seropositive cows were detected in dairy, beef, and mixed-purpose farms. The results of an epidemiological questionnaire showed that no risk factor analyzed was positively associated with seropositivity to VACV. There was no significant association between type of milking (manual/mechanic) and seropositivity to VACV; however, most seropositive cows were present in farms with high daily milk production and high number of lactating and adult cows. Our results indicate that VACV circulates in many regions of DF with considerable prevalence in dairy cows. Control measures to restrict VACV circulation and consequences of the infection may be advisable.

Most milk replacers (MR) contain more lactose compared with whole milk, which, when fed at a large meal size, could influence gut barrier function in calves. This study evaluated how replacing lactose in MR with fat (on a wt/wt basis) affects intestinal histomorphology and permeability in neonatal dairy calves. Thirty-four Holstein-Friesian bull calves were blocked by dam parity and randomly assigned to 1 of 2 treatments (n = 17): a high-lactose (46.1% lactose, 18.0% crude fat, and 23.9% crude protein of dry matter) or a high-fat MR (HF; 39.9% lactose, 24.6% crude fat, and 24.0% crude protein of dry matter). Calves were individually housed and fed pooled colostrum at 1.5 h and 12 h postnatally, at 18 and 9% of metabolic body weight (BW0.75), respectively. From 24 h postnatally until the end of the study (d 7), calves were transitioned to be fed MR (prepared at 15% solids) at 18% of BW0.75 twice daily at 0700 and 1900 h. During postprandial sampling on d 6, intestinal permeability was assessed by mixing lactulose (1.03 g/kg of BW0.75) and d-mannitol (0.31 g/kg of BW0.75) into the morning meal without altering total meal volume. Sequential blood samples were collected via jugular catheter, and total urine was collected for 12 h to measure the marker content. Calves were euthanized 3 h after the morning meal on d 7, and gastrointestinal tract tissues and digesta were collected for analysis of histomorphology, digesta osmolality, and gene expression. The empty gastrointestinal tracts of HF calves were heavier, although length did not differ and differences in histomorphology were minor. Digesta osmolality changed along the tract without differences between treatments. Plasma lactulose was greater in HF, although plasma d-mannitol and the recovery of both markers in urine were unaffected. No significant differences were detected in gene expression, although HF calves tended to have lower expression of TJP1 and CLDN2 and higher expression of proinflammatory cytokine IL1B in ileum tissue. In conclusion, partially replacing lactose in MR with fat resulted in a heavier and more permeable gut, with minor histomorphological differences.

Single intradermal injections of nerve growth factor (NGF) evoke prolonged but temporally distinct sensitization patterns to somatosensory stimuli. Focal administration of the non-histaminergic pruritogen cowhage but not histamine resulted in elevated itch at day 21 after NGF administration. Here, we injected bovine adrenal medulla peptide 8–22 (BAM8–22), β-alanine (β-ALA) and endothelin-1 (ET-1) into NGF-treated skin of 11 healthy volunteers and investigated the corresponding itch/pain and flare reactions. β-ALA was the weakest pruritogen, while BAM8–22 and ET-1 were equally potent as histamine. NGF did not sensitize itch or flare reactions induced by any compound, but injection and evoked pain were increased at day 21 and 49. The involvement of histamine H1 receptors in itch was explored in eight subjects after oral cetirizine. ET-1-induced itch and flare were significantly reduced. BAM8–22 and β-ALA itch were not affected, but flare responses after BAM8–22 reduced by 50%. The results indicate that a single NGF injection does not sensitize for experimentally induced itch but increases pain upon pruritogen injection. In healthy humans, pruritic and algetic processing appear differentially regulated by NGF. However, in patients suffering chronic itch, prolonged elevation of NGF-levels under inflammatory conditions may contribute to elevated itch.

In recent years, due to the shortage of donkey skin resources, the price of Asini Corii Colla has seen a rapid increase. Consequently, fake gelatin prepared from horse, mules, pig, and cow skin has appeared in the market, resulting in unreliable quality of Asini Corii Colla. Therefore, there is an urgent need to develop an efficient and accurate method for improving the quality of Asini Corii Colla. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to determine the donkey skin components in Asini Corii Colla. Accordingly, 0. l g of the evenly mixed sample was weighed and placed in a 50 mL volumetric flask; then, 1% ammonium bicarbonate solution was added to dissolve the sample, and the solution was diluted to the scale. Precisely 1.00 mL of the solution was extracted into a 5 mL volumetric flask, followed by the addition of 1.0 mL trypsin solution and 100 μL mixed internal standard working solution. This mixture was diluted to the scale using 1% ammonium bicarbonate solution, shaken, and placed in an incubator for 16 h to induce enzymolysis at a constant temperature of 37 ℃. The mixture was subsequently removed from the incubator, cooled to ambient temperature, filtered through a 0.22 μm membrane, and analyzed by LC-MS. Separation was performed on an UPLC system with a BEH C18 column (100 mm×2.1 mm, 2.5 μm) under gradient elution using acetonitrile containing 0.1% (v/v) formic acid (B) and water containing 0.1% (v/v) formic acid (A) as the mobile phases at a flow rate of 0.3 mL/min. The column temperature was 30 ℃, and the sample size was 2 μL. The gradient elution conditions are: 0-1 min, 10%B; 1-5 min, 10%B-30%B; 5-5.1 min, 30%B-70%B; 5.1-7 min, 70%B; 7-7.1 min, 70%B-10%B; 7.1-10 min, 10%B. The marker peptides were determined in positive electrospray ionization (ESI +) and multiple reaction monitoring (MRM) modes using the isotopic internal standard method. The optimized enzymolysis conditions were as follows: enzymolysis temperature, 37 ℃; enzymolysis time, 16 h; and amount of enzyme, 1 mL. The two marker peptides showed good linearities in the range of 50 to 1250 mg/L; the correlation coefficients (r) were greater than 0.996, and the limits of quantitation (S/N=10) were 20 mg/kg. At spiked levels of 300 mg/kg, 600 mg/kg, and 900 mg/kg, the average recovery ratios of the two marker peptides were 103.2% to 108.3%, while the relative standard deviations (RSDs) of 1.0%-3.0%. This method was favorable for testing actual samples. Asini Corii Colla from 29 production companies was detected by this method, and the sum contents of the two marker peptides was different because the production process and raw materials were different. The sum contents of the samples were 0.096% to 0.180% with an average of 0.151%. The developed method is simple, reliable, and reproducible, and it is suitable for detecting the donkey hide components Asini Corii Colla.

The continuous trend for a narrowing margin between feed cost and milk prices across dairy farms in the United States highlights the need to improve and maintain feed efficiency. Yeast culture products are alternative supplements that have been evaluated in terms of milk performance and feed efficiency; however, less is known about their potential effects on altering rumen microbial populations and consequently rumen fermentation. Therefore, the objective of this study was to evaluate the effect of yeast culture supplementation on lactation performance, rumen fermentation profile, and abundance of major species of ruminal bacteria in lactating dairy cows. Forty mid-lactation Holstein dairy cows (121 ± 43 days in milk; mean ± standard deviation; 32 multiparous and 8 primiparous) were used in a randomized complete block design with a 7-d adaptation period followed by a 60-d treatment period. Cows were blocked by parity, days in milk, and previous lactation milk yield and assigned to a basal total mixed ration (TMR; 1.6 Mcal/kg of dry matter, 14.6% crude protein, 21.5% starch, and 38.4% neutral detergent fiber) plus 114 g/d of ground corn (CON; n = 20) or basal TMR plus 100 g/d of ground corn and 14 g/d of yeast culture (YC; n = 20; Culture Classic HD, Cellerate Yeast Solutions, Phibro Animal Health Corp.). Treatments were top-dressed over the TMR once a day. Cows were individually fed 1 × /d throughout the trial. Blood and rumen fluid samples were collected in a subset of cows (n = 10/treatment) at 0, 30, and 60 d of the treatment period. Rumen fluid sampled via esophageal tubing was analyzed for ammonia-N, volatile fatty acids (VFA), and ruminal bacteria populations via quantitative PCR amplification of 16S ribosomal DNA genes. Milk yield was not affected by treatment effects. Energy balance was lower in YC cows than CON, which was partially explain by the trend for lower dry matter intake as % body weight in YC cows than CON. Cows fed YC had greater overall ruminal pH and greater total VFA (mM) at 60 d of treatment period. There was a contrasting greater molar proportion of isovalerate and lower acetate proportion in YC-fed cows compared with CON cows. Although the ruminal abundance of specific fiber-digesting bacteria, including Eubacterium ruminantium and Ruminococcus flavefaciens, was increased in YC cows, others such as Fibrobacter succinogenes were decreased. The abundance of amylolytic bacteria such as Ruminobacter amylophilus and Succinimonas amylolytica were decreased in YC cows than CON. Our results indicate that the yeast culture supplementation seems to promote some specific fiber-digesting bacteria while decreasing amylolytic bacteria, which might have partially promoted more neutral rumen pH, greater total VFA, and isovalerate.

The ECM of cartilage is composed of proteoglycans (PG) that contain glycosaminoglycan (GAG), aggrecan, hyaluronic acid (HA) and other molecular components which play an important role in regulating chondrocyte functions via cell-matrix interactions, integrin-mediated signalling etc. Implantation of chondrocytes encapsulated in scaffolds that mimic the micro-architecture of proteoglycan, is expected to enhance cartilage repair. With an aim to create a hydrogel having macromolecular structure that resembles the cartilage-specific ECM, we constructed a hierarchal structure that mimic the PG. The bottle brush structure of the aggrecan was obtained using chondroitin sulphate and carboxymethyl cellulose which served as GAG and core protein mimic respectively. A proteoglycan-like structure was obtained by cross-linking it with modified chitosan that served as a HA substitute. The physico-chemical characteristics of the above cross-linked injectable hydrogel supported long term human articular chondrocyte subsistence and excellent post-injection viability. The chondrocytes encapsulated in the PMH expressed significant levels of articular cartilage specific markers like collagen II, aggrecan, GAGs etc., indicating the ability of the hydrogel to support chondrocyte differentiation. The biocompatibility and biodegradability of the hydrogels was confirmed using suitable in vivo studies. The results revealed that the PG-mimetic hydrogel could serve as a promising scaffold for chondrocyte implantation.

The value of milk hinges on its physicochemical functionality under processing conditions. We examined composition-functionality relationships with individual milks from 24 New Zealand dairy cows, sampled at 3 times over the season. Milks were classified into type A or B, according to the shape of 3-point heat coagulation time versus pH profiles. Milk type changed over the season for half of the cows in the study. Best subsets regression suggested that different factors controlled heat stability in the 2 milk types. Urea concentration was key for both types, but for type A milks, osmotic pressure and milk solids were the most important predictors of heat stability, whereas casein micelle size and ionic calcium predicted heat stability for type B milks. This study revealed that milk type is prone to change over the season, and the findings suggest that optimizing heat stability could be achieved by different means for type A versus type B milks.

Enteric fermentation from ruminants is a primary source of anthropogenic methane emission. This study aims to add another approach for methane mitigation by manipulation of the rumen microbiome. Effects of choline supplementation on methane formation were quantified in vitro using the Rumen Simulation Technique. Supplementing 200 mM of choline chloride or choline bicarbonate reduced methane emissions by 97–100% after 15 days. Associated with the reduction of methane formation, metabolomics analysis revealed high post-treatment concentrations of ethanol, which likely served as a major hydrogen sink. Metagenome sequencing showed that the methanogen community was almost entirely lost, and choline-utilizing bacteria that can produce either lactate, ethanol or formate as hydrogen sinks were enriched. The taxa most strongly associated with methane mitigation were Megasphaera elsdenii and Denitrobacterium detoxificans, both capable of consuming lactate, which is an intermediate product and hydrogen sink. Accordingly, choline metabolism promoted the capability of bacteria to utilize alternative hydrogen sinks leading to a decline of hydrogen as a substrate for methane formation. However, fermentation of fibre and total organic matter could not be fully maintained with choline supplementation, while amino acid deamination and ethanolamine catabolism produced excessive ammonia, which would reduce feed efficiency and adversely affect live animal performance., Scientific Reports, 11 (1), ISSN:2045-2322

Objectives were to determine the effects of 3 levels of dietary cation-anion difference (DCAD) fed prepartum to nulliparous cows on productive and reproductive performance. We enrolled 132 pregnant nulliparous Holstein cows at 250 (248-253) d of gestation in a randomized block design. Cows were blocked by genomic merit of energy-corrected milk yield and assigned randomly to diets varying in DCAD, +200 (P200; n = 43), -50 (N50; n = 45), or -150 (N150; n = 44) mEq/kg of dry matter (DM). Dietary treatments were fed during the last 22 d of gestation and, after calving, postpartum cows received the same lactation diet. Productive performance was evaluated for the first 14 wk of lactation, and reproduction was assessed until 305 d postpartum. Intake of DM prepartum decreased linearly (results presented in sequence as least squares means ± standard error of the mean, P200 vs. N50 vs. N150) with a reduction in DCAD (9.0 vs. 8.9 vs. 8.4 ± 0.1 kg/d), which resulted in linear decreases in net energy balance (0.34 vs. 0.20 vs. -0.36 ± 0.20 Mcal/d), body weight change (1.1 vs. 0.8 vs. 0.3 ± 0.1 kg/d), and mean body weight (652 vs. 649 vs. 643 ± 2 kg) prepartum. Treatment did not affect yield of colostrum (6.3 vs. 5.8 vs. 5.1 ± 0.6 kg) or the contents or yields of fat, protein, lactose, IgG, Ca, or Mg in colostrum. Intake of DM (19.4 vs. 19.2 vs. 19.0 ± 0.2 kg/d), yields of milk (36.6 vs. 36.7 vs. 35.8 ± 0.6 kg/d) or energy-corrected milk (36.7 vs. 36.3 vs. 35.9 ± 0.5 kg/d), feed efficiency (1.93 vs. 1.92 vs. 1.93 ± 0.03 kg of energy-corrected milk per kilogram of DM intake), and content and yield of milk components did not differ among treatments during the first 14 wk of lactation. Prepartum DCAD did not affect the cumulative milk yield by 305 d of lactation (9,653 vs. 10,005 vs. 9,918 ± 196 kg). Of the 132 cows, 40 P200, 45 N50, and 43 N150 received at least 1 artificial insemination (AI), and treatment did not affect pregnancy per AI at first (32.5 vs. 35.6 vs. 37.2%) or all AI (30.6 vs. 33.9 vs. 40.2%), although reducing the DCAD increased the proportion of cows pregnant by 305 d postpartum (76.7 vs. 88.9 vs. 93.2%) without altering the rate of pregnancy. Collectively, manipulating the DCAD of prepartum diets, from +200 to -150 mEq/kg of DM, fed to late gestation nulliparous cows did not affect subsequent lactation productive performance, but may have provided some benefit to reproduction, which warrants further confirmation.

Chitin, an abundant biopolymer on Earth, represents a resource for sustainable functional materials. However, traditional β-chitin production methods involve alkaline treatment at approximately 90 °C for its separation from the protein, thus not suitable as a functional peptide, as it is mixed with an alkaline aqueous solution. This study examined the conversion of squid pen into solid β-chitin and water-soluble peptides using only water at temperatures of 150–250 °C for 30–120 min. Solid β-chitin was converted to its nanofiber form and the physicochemical properties of the β-chitin nanofibers were almost the same as those produced by the traditional method. Because this method uses only water, the protein in the squid pen may also be a functional peptide for lowering blood pressure, by inhibiting the Angiotensin-1 converting enzyme. High-temperature water treatment is a promising environment-friendly technique for complete utilization of squid pen components, including β-chitin and protein., Article, International Journal of Biological Macromolecules.189:921-929(2021)

Thyroglobulin is the precursor protein that serves as a substrate for thyroid hormone synthesis. Here, a three-dimensional cryoEM reconstruction of bovine thyroglobulin at an overall resolution of 2.6 Å is presented. The structure facilitates the definitive identification of thyroxine (acceptor) and dehydroalanine (donor) sites at two of the most evolutionarily conserved sites of hormone formation in vertebrates., Thyroglobulin is a homodimeric glycoprotein that is essential for the generation of thyroid hormones in vertebrates. Upon secretion into the lumen of follicles in the thyroid gland, tyrosine residues within the protein become iodinated to produce monoiodotyrosine (MIT) and diiodotyrosine (DIT). A subset of evolutionarily conserved pairs of DIT (and MIT) residues can then engage in oxidative coupling reactions that yield either thyroxine (T4; produced from coupling of a DIT ‘acceptor’ with a DIT ‘donor’) or triiodothyronine (T3; produced from coupling of a DIT acceptor with an MIT donor). Although multiple iodotyrosine residues have been identified as potential donors and acceptors, the specificity and structural context of the pairings (i.e. which donor is paired with which acceptor) have remained unclear. Here, single-particle cryogenic electron microscopy (cryoEM) was used to generate a high-resolution reconstruction of bovine thyroglobulin (2.3 Å resolution in the core region and 2.6 Å overall), allowing the structural characterization of two post-reaction acceptor–donor pairs as well as tyrosine residues modified as MIT and DIT. A substantial spatial separation between donor Tyr149 and acceptor Tyr24 was observed, suggesting that for thyroxine synthesis significant peptide motion is required for coupling at the evolutionarily conserved thyroglobulin amino-terminus.

BACKGROUND The abnormal ultimate pH (pHu ) in postmortem muscles affect the meat quality and results in substantial economic losses. Dark, firm, and dry (DFD) meat linked with the higher postmortem pHu values and exhibited many quality issues such as dark color, tough texture and shorter shelf life. This research aimed to investigate the effect of protein phosphorylation on variations in beef pHu in order to explore the possible mechanisms underlying DFD meat formation. RESULTS Glycogen and lactate contents were higher, while L* and a* were lower in high pHu beef. Shear force was higher in intermediate pHu group. Global phosphorylation of sarcoplasmic proteins was higher in low pHu samples on day 1 and of myofibrillar proteins was higher in intermediate pHu meat on days 1 and 2 postmortem. Sarcoplasmic protein bands with different phosphorylation levels were identified as containing some glycometabolism and stress response proteins and phosphorylated myofibrillar protein bands were identified sarcomeric and metabolic proteins. CONCLUSIONS Phosphorylation of multiple proteins of glycolytic pathway and contractile machinery may play critical roles in development of DFD beef. © 2021 Society of Chemical Industry.