Milk Citrate Research Articles

Weighed ssGBLUP enhances the genomic prediction accuracy for milk citrate predicted by milk mid-infrared spectra of Holstein cows in early lactation

Previous studies have shown that milk citrate predicted by milk mid-infrared spectra (MIR) is strongly affected by a few genomic regions. This study aimed to explore the effect of the weighted single-step GBLUP on the accuracy of genomic prediction (GP) for MIR-predicted milk citrate in early lactation Holstein cows. A total of 134,517 test-day predicted milk citrate collected within the first 50 d in milk on 52,198 Holstein cows from the first 5 parities were used. There were 122,218 animals in the pedigree, of them 4,479 had genotypic data for 566,170 SNPs. Two data sets (partial and whole data sets) were used to verify whether the accuracy of GP is improved using the following different methods. The (genomic) estimated breeding values (EBV or GEBV) in the partial and whole data sets were estimated by pedigree-based BLUP (ABLUP), single-step GBLUP (ssGBLUP, pedigree-genomic combined using no weight for SNP), weighted ssGBLUP (WssGBLUP, pedigree-genomic combined using weighted SNP), respectively. The difference between the 2 data sets is that the phenotypic data from 2017 to 2019 in the partial data set were set as missing values. 181 youngest cows with genomic were selected as the validation population. A linear regression method was used to compare EBV (GEBV) predicted for partial and whole data sets. The accuracies of GP for ABLUP and ssGBLUP were 0.42 and 0.70, respectively. The accuracies of GP for WssGBLUP in the 5 iterations with different CT (constant) values (determines departure from normality for SNP effects) ranged from 0.70 to 0.86. This study showed that weighted SNP is beneficial in improving prediction accuracy for predicted milk citrate.

Genetic analysis of milk citrate predicted by milk mid-infrared spectra of Holstein cows in early lactation

Milk citrate is regarded as an early biomarker of negative energy balance in dairy cows during early lactation and serves as a suitable candidate phenotype for genomic selection due to its wide availability across a large number of cows through milk mid-infrared spectra prediction. However, its genetic background is not well known. Therefore, the objectives of this study were to (1) analyze the genetic parameters of milk citrate; (2) identify genomic regions associated with milk citrate; and (3) analyze the functional annotation of candidate genes and quantitative trait loci (QTL) related to milk citrate in Walloon Holstein cows. In total, 134,517 test-day milk-citrate phenotypes (mmol/L) collected within the first 50 d in milk on 52,198 Holstein cows were used. These milk-citrate phenotypes, predicted by milk mid-infrared spectra, were divided into 3 traits according to the first (citrate1), second (citrate2), and third to fifth parity (citrate3+). Genomic information for 566,170 SNPs was available for 4,479 animals. A multiple-trait repeatability model was used to estimate genetic parameters. A single-step GWAS was used to identify candidate genes for citrate and post-GWAS analysis was done to investigate the relationship and function of the identified candidate genes. The heritabilities estimated for citrate1, citrate2, and citrate3+ were 0.40, 0.37, and 0.35, respectively. The genetic correlations among the 3 traits ranged from 0.98 to 0.99. The genomic correlations among the 3 traits were also close to 1.00 across the genomic regions (1 Mb) in the whole genome, which means that citrate can be considered as a single trait in the first 5 parities. In total, 603 significant SNPs located on 3 genomic regions (chromosome 7, 68.569-68.575 Mb; chromosome 14, 0.15-1.90 Mb; and chromosome 20, 54.00-64.28 Mb), were identified to be associated with milk citrate. We identified 89 candidate genes including GPT, ANKH, PPP1R16A, and 32 QTL reported in the literature related to the identified significant SNPs. These identified QTL were mainly reported associated with milk fatty acids and metabolic diseases in dairy cows. This study suggests that milk citrate in Holstein cows is highly heritable and has the potential to be used as an early proxy for the negative energy balance of Holstein cows in a breeding objective.

Role of Trisodium Citrate and Nanominerals in Mastitis Management in Dairy Animals: A Review

Mastitis is the most common disease of dairy cattle across the globe including India and cause huge economic losses to dairy farmers. Mastitis is also the most common reason for antibiotic usage in dairy animals, which is an important reason for antimicrobial resistance development in human and veterinary important pathogen. Optimization of nutrition and improvement of immunity of lactating cows are important strategies to reduce the susceptibility to mastitis and thereby reduction of antibiotic use. Copper, Zinc and Manganese are important trace minerals for maintaining udder health and immunity but, their contents and bioavailability is affected by several factors. Citrate, the precursor for milk synthesis is an important determinant of udder health but, the role of citrate in udder health is not clear as milk citrate level is affected by various factors. This review discusses about the role of trisodium citrate and various forms of trace minerals in mastitis management in dairy animals.

Phenotype testing, genome analysis, and metabolic interactions of three lactic acid bacteria strains existing as a consortium in a naturally fermented milk.

This work reports the characterization of three lactic acid bacteria (LAB) strains –Lactococcus lactis LA1, Lactococcus cremoris LA10, and Lactiplantibacillus plantarum LA30– existing as a stable consortium in a backslopping-inoculated, naturally fermented milk (NFM). This study aimed at uncovering the biochemical and genetic basis of the stability of the consortium and the cooperativity among the strains during milk fermentation. All three strains were subjected to phenotyping, covering the utilization of carbohydrates, enzyme activity, and antibiotic resistance. The strains were grown in milk individually, as well as in all possible combinations, and the resulting fermented product was analyzed for sugars, organic acids, and volatile compounds. Finally, the genomes of the three strains were sequenced and analyzed for genes associated with technological and safety properties. As expected, wide phenotypic diversity was seen between the strains. Lactococcus cremoris LA10 was the only strain to reach high cell densities and coagulate milk alone after incubation at 22°C for 24 h; congruently, it possessed a gene coding for a PrtP type II caseinolytic protease. Compared to any other fermentation, acetaldehyde concentrations were greater by a factor of six when all three strains grew together in milk, suggesting that its production might be the result of an interaction between them. Lactococcus lactis LA1, which carried a plasmid-encoded citQRP operon, was able to utilize milk citrate producing diacetyl and acetoin. No genes encoding virulence traits or pathogenicity factors were identified in any of the strains, and none produced biogenic amines from amino acid precursors, suggesting them to be safe. Lactiplantibacillus plantarum LA30 was susceptible to tetracycline, although it harbors a disrupted antibiotic resistance gene belonging to the tetM/tetW/tetO/tetS family. All three strains contained large numbers of pseudogenes, suggesting that they are well adapted (“domesticated”) to the milk environment. The consortium as a whole or its individual strains might have a use as a starter or as starter components for dairy fermentations. The study of simple consortia, such as that existing in this NFM, can help reveal how microorganisms interact with one another, and what influence they may have on the sensorial properties of fermented products.

Isolation and phenotypic and genomic characterization of Tetragenococcus spp. from two Spanish traditional blue-veined cheeses made of raw milk

High throughput sequencing has recently revealed the presence of Tetragenococcus-related DNA sequences in dairy environments such as brine and cheeses. In the present work, a selective medium was developed to isolate Tetragenococcus spp. from two ripened, traditional, Spanish, blue-veined cheese varieties made from raw milk. The strains recovered belonged to either Tetragenococcus koreensis or Tetragenococcus halophilus species. Twenty of these isolates (15 of T. koreensis and 5 of T. halophilus) were then subjected to a battery of phenotypic and genetic tests, and six strains (4 T. koreensis and 2 T. halophilus) to genome sequencing. Wide genetic and phenotypic diversity was noted. All strains grew poorly in milk, producing small quantities of lactic and acetic acids. Most strains used lactose as a carbon source and ferment milk citrate. In agreement, genome analysis detected in the genome of the six strains analyzed gene clusters harboring several lactose/galactose-related genes and genes encoding citrate metabolic enzymes (permease, citrate lyase, and oxaloacetate decarboxylase). Most of the tested strains were resistant to erythromycin and clindamycin, and a few to other antimicrobial agents, but neither known mutations nor acquired genes conferring resistance to antibiotics were identified in their genomes. Neither were genes coding for pathogenicity or virulence factors detected. Decarboxylase-encoding genes involved in biogenic amine production were not identified, in keeping with the strains' negative biogenic amine-producer phenotype. Genome comparison revealed vast arrays of genes (similar in number to those described in other lactic acid bacteria) coding for components of proteolytic and lipolytic systems. Tetragenococcus strains showing desirable traits plus the absence of detrimental features might be exploitable in the form of secondary, adjunct or ripening cultures to ensure the typical bouquet of traditional blue-veined cheeses is obtained, or to diversify the final flavor in other varieties.

Effects of Protein, Calcium, and pH on Gene Transcription, Cell-Envelope Peptidase Activity of Lactococcus lactis Strains, and the Formation of Bitter Peptides.

Calcium- and protein-rich fermented milk products, such as concentrated yoghurts and fresh cheeses, may contain undesired bitter peptides, which are generated by the proteolytic cleavage of casein. Up to now, it is not clear whether this process is caused by endogenous milk enzymes, such as plasmin and cathepsin D, or whether proteolytic enzymes from applied starter cultures, such as the lactococcal cell-envelope peptidase PrtP, are involved. A sensory analysis of fresh cheese products made from milk concentrates fermented with prtP-negative and -positive Lactococcus lactis strains revealed bitterness in the products fermented with prtP-positive L. lactis strains. Two prtP-positive strains, LTH 7122 and LTH 7123, were selected to investigate the effect of increased calcium concentrations (additional 5 mM and 50 mM CaCl2) at neutral (pH 6.6) and acidic (pH 5.5) pH-values on the transcription of the prtP gene and its corresponding PrtP peptidase activity in milk citrate broth (MCB). For both strains, it was shown that prtP transcription was upregulated only under slightly elevated calcium conditions (5 mM CaCl2) after 5 h of growth. In concordance with these findings, PrtP peptidase activity also increased. When higher concentrations of calcium were used (50 mM), prtP expression of both strains decreased strongly by more than 50%. Moreover, PrtP peptidase activity of strain LTH 7123 decreased by 15%, but enzymatic activity of strain LTH 7122 increased slightly during growth under elevated calcium concentrations (50 mM CaCl2). Fermentations of reconstituted casein medium with 3.4% (w/v) and 8.5% (w/v) protein and different calcium concentrations using strain LTH 7122 revealed no clear relationship between prtP transcription and calcium or protein concentration. However, an increase in PrtP peptidase activity under elevated protein and calcium conditions was observed. The activity increase was accompanied by increased levels of bitter peptides derived from different casein fractions. These findings could be a possible explanation for the bitterness in fermented milk concentrates that was detected by a trained bitter panel.

Metabolomics Approach Explore Diagnostic Biomarkers and Metabolic Changes in Heat-Stressed Dairy Cows.

Simple SummaryHeat stress results in a decline in the production performance of the dairy cows. This study explored the metabolomes of milk and blood plasma in heat-stressed cows by means of 1H nuclear magnetic resonance (1H NMR)-based metabolomics tools. Findings of the current experiment reveal that HS alters the metabolic composition of milk and blood plasma in lactating dairy cows. In brief, HS alters twelve metabolites in blood plasma and eight metabolites in milk, which are primarily involved in proteolysis, gluconeogenesis, and milk fatty acid synthesis, suggesting that these metabolites could be possible biomarkers for heat-stressed dairy cows.In the present experiment, we investigated the impact of heat stress (HS) on physiological parameters, dry matter intake, milk production, the metabolome of milk, and blood plasma in lactating Holstein dairy cows. For this purpose, 20 Holstein lactating cows were distributed in two groups in such a way that each group had 10 cows. A group of 10 cows was reared in HS conditions, while the other group of 10 cows was reared in the thermoneutral zone. The results of the experiment showed that cows subjected to HS had higher respiration rates (p < 0.01) and greater rectal temperature (p < 0.01). Results of milk production and composition explored that HS lowered milk production (p < 0.01) and milk protein percentage (p < 0.05) than cows raised in a thermoneutral place. Furthermore, HS increased the concentrations of N-acetyl glycoprotein, scyllo-inositol, choline, and pyridoxamine in milk, while HS decreased the concentrations of O-acetyl glycoprotein, glycerophosphorylcholine, citrate, and methyl phosphate in milk. Moreover, HS enhanced plasma concentrations of alanine, glucose, glutamate, urea, 1-methylhistidine, histidine, and formate in cows, while the plasma concentration of low-density lipoprotein, very-low-density lipoprotein, leucine, lipid, and 3-hydroxybutyrate decreased due to HS. Based on the findings of the current research, it is concluded that HS alters the milk and blood plasma metabolites of lactating Holstein dairy cows. Overall, in the current experiment, HS altered eight metabolites in milk and twelve metabolites in the plasma of lactating Holstein dairy cows. Furthermore, the current study explored that these metabolites were mainly involved in proteolysis, gluconeogenesis, and milk fatty acid synthesis and could be potential biomarkers for dairy cows undergoing HS.

Mother's Own Milk Biomarkers Predict Coming to Volume in Pump-Dependent Mothers of Preterm Infants

To assess serial secretory activation biomarker concentrations (sodium [Na], potassium [K], Na:K, protein, lactose, and citrate) in mother's own milk (MOM) from breast pump-dependent mothers of preterm infants to determine associations with coming to volume (CTV), defined as producing at least 500mL/day MOM by day 14 postpartum. We collected serial MOM samples and pumped MOM volume data for 14days postpartum in mothers who delivered at <33weeks of gestation. Regression models and the Mann-Whitney U test were used to evaluate associations. Among 40 mothers, 39 (mean gestational age, 28.8weeks; 67% overweight/obese; 59% nonwhite) had paired MOM volume and biomarker data; 33% achieved CTV between postpartum days 6 and 14. In univariate models, MOM Na on postpartum day 5 and Na:K on days 3 and 5 were associated with CTV. Mothers achieving CTV were more likely to have postpartum Na:K ≤1 on day 3 (75% vs 25%; P = .06) and ≤0.8 on day 5 (69% vs 10%; P<.01). In a multivariable regression model, day 5 Na:K (1 unit decrease in Na:K: OR, 18.7; 95% CI, 1.13-311.41; P=.049) and maternal prepregnancy body mass index (BMI) (1 unit increase in BMI: OR, 0.88; 95% CI, 0.78-0.99; P=.04) were associated with CTV between postpartum days 6 and14. Secretory activation and CTV were compromised in breast pump-dependent mothers with preterm delivery. CTV was predicted by MOM Na level and Na:K. These biomarkers have potential as objective point-of-care measures to detect potentially modifiable lactation problems in a high-risk population.

Effects of abomasal infusion of essential fatty acids together with conjugated linoleic acid in late and early lactation on performance, milk and body composition, and plasma metabolites in dairy cows

Rations including high amounts of corn silage are currently very common in dairy production. Diets with corn silage as forage source result in a low supply of essential fatty acids, such as α-linolenic acid, and may lead to low conjugated linoleic acid (CLA) production. The present study investigated the effects of abomasal infusion of essential fatty acids, especially α-linolenic acid, and CLA in dairy cows fed a corn silage-based diet on performance, milk composition, including fatty acid (FA) pattern, and lipid metabolism from late to early lactation. Rumen-cannulated Holstein cows (n = 40) were studied from wk 9 antepartum to wk 9 postpartum and dried off 6 wk before calving. The cows were assigned to 1 of 4 treatment groups. Cows were abomasally supplemented with coconut oil (CTRL, 76 g/d), linseed and safflower oil (EFA, 78 and 4 g/d; linseed/safflower oil ratio = 19.5:1; n-6/n-3 FA ratio = 1:3), Lutalin (CLA, 38 g/d; BASF SE, Ludwigshafen, Germany; isomers cis-9,trans-11 and trans-10,cis-12 each 10 g/d) or EFA+CLA. Milk composition was analyzed weekly, and blood samples were taken several times before and after parturition to determine plasma concentrations of metabolites related to lipid metabolism. Liver samples were obtained by biopsy on d 63 and 21 antepartum and on d 1, 28, and 63 postpartum to measure triglyceride concentration. Body composition was determined after slaughter. Supplementation of CLA reduced milk fat concentration, increased body fat mass, and improved energy balance (EB) in late and early lactation, but EB was lowest during late lactation in the EFA group. Cows with CLA treatment alone showed an elevated milk citrate concentration in early lactation, whereas EFA+CLA did not reveal higher milk citrate but did have increased acetone. Milk protein was increased in late lactation but was decreased in wk 1 postpartum in CLA and EFA+CLA. Milk urea was reduced by CLA treatment during the whole period. After calving, the increase of nonesterified fatty acids in plasma was less in CLA groups; liver triglycerides were raised lowest at d 28 in CLA groups. Our data confirm an improved metabolic status with CLA but not with exclusive EFA supplementation during early lactation. Increased milk citrate concentration in CLA cows points to reduced de novo FA synthesis in the mammary gland, but milk citrate was less affected in EFA+CLA cows, indicating that EFA supplementation may influence changes in mammary gland FA metabolism achieved by CLA.

From Waste to Taste-Efficient Production of the Butter Aroma Compound Acetoin from Low-Value Dairy Side Streams Using a Natural (Nonengineered) Lactococcus lactis Dairy Isolate.

Lactococcus lactis subsp. lactis biovar diacetylactis is widely used in dairy fermentations as it can form the butter aroma compounds acetoin and diacetyl from citrate in milk. Here, we explore the possibility of producing acetoin from the more abundant lactose. Starting from a dairy isolate of L. lactis biovar diacetylactis, we obtained a series of mutants with low lactate dehydrogenase (ldh) activity. One isolate, RD1M5, only had a single insertion mutation in the ldh gene compared to its parental strain as revealed by whole genome resequencing. We tested the ability of RD1M5 to produce acetoin in milk. With aeration, all the lactose could be consumed, and the only product was acetoin. In a simulated cheese fermentation, a 50% increase in acetoin concentration could be achieved. RD1M5 turned out to be an excellent cell factory for acetoin and was able to convert lactose in dairy waste into acetoin with high titer (41 g/L) and high yield (above 90% of the theoretical yield). Summing up, RD1M5 was found to be highly robust and to grow excellently in milk or dairy waste. Being natural in origin opens up for applications within dairies as well as for safe production of food-grade acetoin from low-cost substrates.

Short communication: Investigation of the temporal relationships between milk mid-infrared predicted biomarkers and lameness events in later lactation

This study reports on the exploration of temporal relationships between milk mid-infrared predicted biomarkers and lameness events. Lameness in dairy cows is an issue that can vary greatly in severity and is of concern for both producers and consumers. Metabolic disorders are often associated with lameness. However, lameness can arise weeks or even months after the metabolic disorder, making the detection of causality difficult. We already use mid-infrared technology to predict major milk components, such as fat or protein, during routine milk recording and for milk payment. It was recently shown that this technology can also be used to predict novel biomarkers linked to metabolic disorders in cows, such as oleic acid (18:1 cis-9), β-hydroxybutyrate, acetone, and citrate in milk. We used these novel biomarkers as proxies for metabolic issues. Other studies have explored the possibility of using mid-infrared spectra to predict metabolic diseases and found it (potentially) usable for indicating classes of metabolic problems. We wanted to explore the possible relationship between mid-infrared-based metabolites and lameness over the course of lactation. In total, data were recorded from 6,292 cows on 161 farms in Austria. Lameness data were recorded between March 2014 and March 2015 and consisted of 37,555 records. Mid-infrared data were recorded between July and December 2014 and consisted of 9,152 records. Our approach consisted of fitting preadjustments to the data using fixed effects, computing pair-wise correlations, and finally applying polynomial smoothing of the correlations for a given biomarker at a certain month in lactation and the lameness events scored on severity scale from sound or non-lame (lameness score of 1) to severely lame (lameness score of 5) throughout the lactation. The final correlations between biomarkers and lameness scores were significant, but not high. However, for the results of the present study, we should not look at the correlations in terms of absolute values, but rather as indicators of a relationship through time. When doing so, we can see that metabolic problems occurring in mo 1 and 3 seem more linked to long-term effects on hoof and leg health than those in mo 2. However, the quantity (only 1 pair-wise correlation exceeded 1,000 observations) and the quality (due to limited data, no separation according to more metabolic-related diseases could be done) of the data should be improved.

Effects of abomasal infusion of essential fatty acids and conjugated linoleic acid on performance and fatty acid, antioxidative, and inflammatory status in dairy cows

The objective of this study was to test the effects of essential fatty acids (EFA), particularly α-linolenic acid, and conjugated linoleic acid (CLA) supplementation on fatty acid (FA) composition, performance, and systemic and hepatic antioxidative and inflammatory responses in dairy cows. Four cows (126 ± 4 d in milk) were investigated in a 4 × 4 Latin square and were abomasally infused with 1 of the following for 6 wk: (1) coconut oil (control treatment, CTRL; 38.3 g/d; providing saturated FA), (2) linseed and safflower oil (EFA treatment; 39.1 and 1.6 g/d, respectively; providing mainly α-linolenic acid), (3) Lutalin (BASF, Ludwigshafen, Germany; CLA treatment; cis-9,trans-11 and trans-10,cis-12 CLA, 4.6 g/d each), (4) or EFA+CLA. The initial dosage was doubled every 2 wk, resulting in 3 dosages (dosage 1, 2, and 3). Cows were fed a corn silage-based total mixed ration with a high n-6/n-3 FA ratio. Dry matter intake and milk yield were recorded daily, and milk composition was measured weekly. The FA compositions of milk fat and blood plasma were analyzed at wk 0, 2, 4, and 6. The plasma concentration and hepatic mRNA abundance of parameters linked to the antioxidative and inflammatory response were analyzed at wk 0 and 6 of each treatment period. Infused FA increased in blood plasma and milk of the respective treatment groups in a dose-dependent manner. The n-6/n-3 FA ratio in milk fat was higher in CTRL and CLA than in EFA and EFA+CLA. The sum of FA <C16 in milk fat decreased in CLA and EFA+CLA in a dosage-dependent manner. Energy-corrected milk and milk fat decreased in CLA and EFA+CLA in a dosage-dependent manner and were higher in EFA and CTRL than in CLA at dosages 2 and 3. Energy balance tended to be highest in CLA cows. Milk protein content was lower in CLA and EFA+CLA than in CTRL. Milk urea concentration decreased in CLA and EFA+CLA in a dosage-dependent manner and was lower in CLA and EFA+CLA than in EFA and CTRL at dosages 2 and 3. Milk citrate concentration increased in CLA in a dosage-dependent manner and was higher in CLA and EFA+CLA than in EFA and CTRL. Glutathione peroxidase activity in blood plasma was lower in CTRL than in EFA, and plasma concentration of β-carotene increased in EFA and EFA+CLA with dosage. Increased milk citrate pointed at reduced de novo FA synthesis and a better antioxidative status in milk due to CLA treatment. Supplementation with CLA may also affect milk protein synthesis, but EFA and CLA treatment did not influence the inflammatory status in a consistent manner in mid-lactating cows.

Natural variations of citrate and calcium in milk and their effects on milk processing properties

Natural variations among milk constituents, and their relations to each other as well as to processing parameters, represent possibilities for differentiation of milk to produce high-quality natural products. In this study, we focused on natural variations in milk citrate and its interplay with calcium distribution in milk, in relation to processing properties. Milk samples from individual cows from farms varying in feeding and management practices were collected from April to June 2017 to maximize natural variations in citrate and calcium. Chemical composition, rennet coagulation properties, and ethanol stability were analyzed for all milk samples. We focused particularly on calcium distribution and citrate content and the correlation of these to other milk parameters. No significant change in citrate content was observed during the sampling period, which suggests that mechanisms other than feeding affect citrate levels in milk. Several significant correlations were found, including a positive correlation between complexed serum calcium and citrate, and a negative correlation between urea and ionic calcium. These are both of interest in relation to further processing, as with regard to the stability of UHT milk and in cheese making. Although the correlation between complexed serum calcium and citrate may be explained by their affinity, the underlying driver for the negative relationship between natural milk urea and ionic calcium needs to be clarified by further studies. Furthermore, milk from the different farms varied not only with regard to organic versus conventional farming systems; feeding practices between farms also play an important role in milk composition and functionality. However, none of the differences in milk composition between farms were found to decrease milk functionality and thus would probably not cause any processing problems.

Mineral and Citrate Concentrations in Milk Are Affected by Seasons, Stage of Lactation and Management Practices

This study was conducted to examine associations between nutrition, time of year and season of calving on milk mineral concentrations in 24 pasture-based dairy farms. There was substantial variation in the concentrations (mean with range in parentheses) of Ca 1072 (864–1310) mg/kg; citrate 1579 (880–2080) mg/kg; P 885 (640–1040) mg/kg; Mg 98 (73–122) mg/kg; Na 347 (248–554) mg/kg; K 1534 (1250–2010) mg/kg; and S 295 (155–372) mg/kg with most of the variation associated with stage of lactation, although the influence of days in milk was different for different minerals. Feeding practices were also important in determining the concentrations of some components. Milk Ca, citrate, P, and K concentrations were greater (p &lt; 0.05) in cows receiving mineral supplements, while Ca, P and Mg were positively correlated (p &lt; 0.05) with the amount of concentrates fed. Milk citrate and K concentrations were positively correlated (p &lt; 0.05) with herbage allowance. Milk Se (15 (&lt;3–37) µg/kg) and Zn (3.4 (2.2–4.9) mg/kg) also varied with time of year, with Se concentrations also greater (13.7 vs 11.6 µg/kg) in spring compared with autumn calving herds and positively correlated (p &lt; 0.05) with the amount of concentrates fed and mineral supplementation. These data indicate that calving pattern and feeding practices could be used to reduce variation in the measured milk mineral concentrations.

Normal Human Lactation: closing the gap.

With the exception of infant growth, there are no well-defined parameters describing normal human lactation. This represents a major gap in the continuum of care that does not exist for other major organs. Biological normality occurs naturally and is characterized by well-integrated function. We have proposed a definition that highlights four key elements that describe parameters for biological normality: comfort, milk supply, infant health, and maternal health. Notwithstanding the current limitations, published data have been collated to provide preliminary markers for the initiation of lactation and to describe objective tests once lactation is established. Reference limits have been calculated for maternal markers of secretory activation, including progesterone in maternal blood and total protein, lactose, sodium, and citrate in maternal milk. Objective measurements for established lactation, including 3-hourly pumping and 24-hour milk production, together with pre-feed to post-feed milk fat changes (a useful indicator of the available milk removed by the infant) have been outlined. Considered together with the parameters describing normal function, this information provides a preliminary objective framework for the assessment of human lactation.

Effects of photoperiod modulation and melatonin feeding around drying-off on bovine mammary gland involution

The risk for a dairy cow to acquire new intramammary infections is high during the transition from lactation to the dry period, because of udder engorgement and altered immune functions. Once the gland is fully involuted, it becomes much more resistant to intramammary infections. Therefore, strategies to depress milk yield before drying-off and accelerate the involution process after drying-off could be beneficial for udder health. The objective of this study was to assess the effect of photoperiod manipulation and melatonin feeding from 14 d before to 14 d after drying-off on the speed of the involution process. Thirty Holstein cows in late lactation were randomly allocated to one of the following treatments: (1) a long-day photoperiod (16 h of light: 8 h of darkness), (2) a short-day photoperiod (8 h of light: 16 h of darkness), and (3) a long-day photoperiod supplemented by melatonin feeding (4 mg/kg of body weight). Milk and blood samples were collected on d -26, -19, -12, -5, -1, 1, 3, 5, 7, 10, and 14 relative to the last milking to determine concentrations of mammary gland involution markers and serum prolactin. Additional blood samples were taken around milking on d -15, before the start of the treatments, and on d -1, before drying-off, to evaluate the treatment effects on milking-induced prolactin release. The short-day photoperiod slightly decreased milk production and basal prolactin secretion during the dry period. The milking-induced prolactin surge was smaller on d -1 than on d -15 regardless of the treatments. Lactoferrin concentration, somatic cell count, and BSA concentration as well as matrix metalloproteinase-2 and -9 activities increased in mammary secretions during the first 2 wk of the dry period, whereas milk citrate concentration and the citrate:lactoferrin molar ratio decreased. The rates of change of these parameters were not significantly affected by the treatments. The long-day photoperiod supplemented by melatonin feeding did not affect milk production, prolactin secretion, or mammary gland involution. Under the conditions in this study, photoperiod modulation and melatonin feeding did not appear to affect the rate of mammary gland involution.

Effects of storage temperature and extension media on motility of caprine spermatozoa

The survivability of caprine spermatozoa was compared in semen extenders prepared from the conventional egg-yolk citrate and goat-milk citrate. Motility was comparable in both sets of extenders during the first 24 hours post-extension when extended ejaculates were stored at either room temperature (28 O C) or refrigerator temperature (5 O C). Motility in goat milk citrate extenders at 5 O C was significantly higher (p<0.05) than that stored at 28 O C and in egg-yolk citrate extenders at both investigated temperature (p<0.05) 48 hours post extension. Beyond 72 hours of storage, motility in both extenders and at both temperature was less than 40%. The results showed that goat milk in part can replace egg-yolk as a medium for semen extension. Therefore, an extender with concentration of 10-20 % heated goat milk, 70-80% sodium citrate and 10% egg-yolk could be used to store goat semen at 5 O C for 2-3 days. Keywords : Egg-yolk citrate, Goat-milk citrate, Motility, Semen extension, Spermatozoa

0218 Optimization of probes and PCR conditions for the correlation between 4 genes and production of high citrate in milk

0218 Optimization of probes and PCR conditions for the correlation between 4 genes and production of high citrate in milk

Determination of freezing point standard for UHT milk

&lt;p&gt;The extended shelf life and undemanding storage requirements of UHT (ultra high temperature) milk make it the most consumed fluid milk in Brazil. The thermal treatment to which UHT milk is subjected alters casein micellar structure, leading to protein sedimentation and age gelation. Therefore, the current legislation allows the addition of up to 0.1% stabilizing salts (sodium citrate and/or sodium phosphates) in milk. The adding of stabilizing salts hinders the possibility of inclusion of freezing point and density as quality parameters of UHT milk. Evaluation of freezing point is the main test to detect adulteration of milk with water; however, addition of stabilizers alters its results. This study aimed to measure the alterations in freezing point and density caused by stabilizers in samples of pasteurized milk added with sodium citrate or sodium phosphates, in order to propose standards for freezing point and density for UHT milk. For this purpose, 16 aliquots of milk were prepared in nine repetitions with different concentrations (0%, 0.001%, 0.01%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15%) of sodium citrate or a commercial mix of sodium mono-, di- and triphosphate. The density and freezing point of all aliquots were tested. As expected, addition of stabilizers lowered the freezing point of milk. Addition of 0.1% citrate and commercial mix promoted an average reduction of –0.021°H (± 0.001) and –0.017°H (± 0.002) in freezing point, respectively. However, no significant change in the density of milk was observed. Given that UHT milk industry usually adds the maximum limit of 0.1% of stabilizers in order to obtain proper effect, the interval from –0.572°H to –0.545°H can be suggested as the standard for freezing point to UHT milk. However, as the addition of stabilizers caused no significant change in the density of UHT milk, the density parameter of raw and pasteurized milk in the range from 1.028 g.mL-1 to 1.034 g.mL-1 could be applied to UHT milk.&lt;/p&gt;

Determination of freezing point standard for UHT milk

&lt;p&gt;The extended shelf life and undemanding storage requirements of UHT (ultra high temperature) milk make it the most consumed fluid milk in Brazil. The thermal treatment to which UHT milk is subjected alters casein micellar structure, leading to protein sedimentation and age gelation. Therefore, the current legislation allows the addition of up to 0.1% stabilizing salts (sodium citrate and/or sodium phosphates) in milk. The adding of stabilizing salts hinders the possibility of inclusion of freezing point and density as quality parameters of UHT milk. Evaluation of freezing point is the main test to detect adulteration of milk with water; however, addition of stabilizers alters its results. This study aimed to measure the alterations in freezing point and density caused by stabilizers in samples of pasteurized milk added with sodium citrate or sodium phosphates, in order to propose standards for freezing point and density for UHT milk. For this purpose, 16 aliquots of milk were prepared in nine repetitions with different concentrations (0%, 0.001%, 0.01%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15%) of sodium citrate or a commercial mix of sodium mono-, di- and triphosphate. The density and freezing point of all aliquots were tested. As expected, addition of stabilizers lowered the freezing point of milk. Addition of 0.1% citrate and commercial mix promoted an average reduction of –0.021°H (± 0.001) and –0.017°H (± 0.002) in freezing point, respectively. However, no significant change in the density of milk was observed. Given that UHT milk industry usually adds the maximum limit of 0.1% of stabilizers in order to obtain proper effect, the interval from –0.572°H to –0.545°H can be suggested as the standard for freezing point to UHT milk. However, as the addition of stabilizers caused no significant change in the density of UHT milk, the density parameter of raw and pasteurized milk in the range from 1.028 g.mL-1 to 1.034 g.mL-1 could be applied to UHT milk.&lt;/p&gt;