Secretion of whey acidic protein and cystatin is down regulated at mid-lactation in the red kangaroo (Macropus rufus)

Milk collected from the red kangaroo (Macropus rufus) between day 100 and 260 of lactation showed major changes in milk composition at around day 200 of lactation, the time at which the pouch young begins to temporarily exit the pouch and eat herbage. The carbohydrate content of milk declined abruptly at this time and although there was only a small increase in total protein content, SDS PAGE analysis of milk revealed asynchrony in the secretory pattern of individual proteins. The levels of alpha-lactalbumin, beta-lactoglobulin, serum albumin and transferrin remain unchanged during lactation. In contrast, the protease inhibitor cystatin, and the putative protease inhibitor whey acidic protein (WAP) first appeared in milk at elevated concentrations after approximately 150 days of lactation and then ceased to be secreted at approximately 200 days. In addition, a major whey protein, late lactation protein, was first detected in milk around the time whey acidic protein and cystatin cease to be secreted and was present at least until day 260 of lactation. The co-ordinated, but asynchronous secretion of putative protease inhibitors in milk may have several roles during lactation including tissue remodelling in the mammary gland and protecting specific proteins in milk required for physiological development of the dependent young.     

Effects of stage of pregnancy on variance components,daily milk yields and 305-day milk yield in Holstein cows,as estimated by using a test-day model

Pregnancy and calving are elements indispensable for dairy production, but the daily milk yield of cows decline as pregnancy progresses, especially during the late stages. Therefore, the effect of stage of pregnancy on daily milk yield must be clarified to accurately estimate the breeding values and lifetime productivity of cows. To improve the genetic evaluation model for daily milk yield and determine the effect of the timing of pregnancy on productivity, we used a test-day model to assess the effects of stage of pregnancy on variance component estimates, daily milk yields and 305-day milk yield during the first three lactations of Holstein cows. Data were 10 646 333 test-day records for the first lactation; 8 222 661 records for the second; and 5 513 039 records for the third. The data were analyzed within each lactation by using three single-trait random regression animal models: one model that did not account for the stage of pregnancy effect and two models that did. The effect of stage of pregnancy on test-day milk yield was included in the model by applying a regression on days pregnant or fitting a separate lactation curve for each days open (days from calving to pregnancy) class (eight levels). Stage of pregnancy did not affect the heritability estimates of daily milk yield, although the additive genetic and permanent environmental variances in late lactation were decreased by accounting for the stage of pregnancy effect. The effects of days pregnant on daily milk yield during late lactation were larger in the second and third lactations than in the first lactation. The rates of reduction of the 305-day milk yield of cows that conceived fewer than 90 days after the second or third calving were significantly (P<0.05) greater than that after the first calving. Therefore, we conclude that differences between the negative effects of early pregnancy in the first, compared with later, lactations should be included when determining the optimal number of days open to maximize lifetime productivity in dairy cows.     

"Whey" proteins of milk of the red (Macropus rufus) and eastern grey (Macropus giganteus) kangaroo

1. A comparison is made of gel electrophoretic patterns of the "whey" proteins of the milk of red (Macropus rufus) and eastern grey (Macropus giganteus) kangaroos at various stages of lactation. Qualitative and quantitative changes occur with time during the mature phase of lactation of both types. Their onset is related solely to the stage of lactation. "Whey" proteins are isolated and characterised and the nature of protein changes determined for the first time. 2. The anodic electrophoretic pattern is divided into 6 main zones (designated A F in order of decreasing mobility) and 2 cathodic zones (G and H) that are only detected in the milk of M. giganteus. 3. Zones A, B and C are milk specific. Zone B is present throughout lactation in both species and is an alpha-lactalbumin. Zones A and C are present only in late lactation, zone C, usually, but not always, appearing first. Zone A is an alpha-lactalbumin in M. giganteus, but is not an alpha-lactalbumin in M. rufus. Zone C appears to be the same protein in both species and is possibly a beta-lactoglobulin. 4. Zone D is kangaroo serum albumin and zone E is possibly a beta 2-microglobulin. Zone F contains three main iron (III) binding bands whose relative intensity varies with stage of lactation. Their intensity differs from the corresponding blood serum transferrin bands. 5. Zone H of Macropus giganteus is a lysozyme. 6. Lactose is present in the milk, but is not the principal sugar. 7. The significance of the results is discussed.     

Milk Composition during Lactation Suggests a Mechanism for Male Biased Allocation of Maternal Resources in the Tammar Wallaby (Macropus eugenii)

Recent research has found empirical evidence in support of the Trivers-Willard Hypothesis that offspring sex allocation is correlated with maternal investment. Tammar wallabies birthing sons have higher investment ability; however a mechanism for sex specific differential allocation of maternal resources in wallabies remains elusive. In metatherians the majority of maternal investment is during lactation. To examine if differential allocation occurs during lactation, we measured total milk protein, lipid and carbohydrates, from mothers with male and female pouch young, during phase 2B (100–215 days post partum) and phase 3 (215–360 days post partum) of lactation. Mothers of sons allocated significantly higher levels of protein than mothers of daughters during phase 2B of lactation, however no sex specific difference in maternal allocation was found for lipids, carbohydrates, or any milk component during phase 3 of lactation. We were unable to measure milk production to establish any differences in the amount of milk allocated. However, with the production of more milk comes a dilution effect on milk components. Given that we find no apparent dilution of milk components may suggest equality in milk production. Offspring body weight at 14 months of age was related to protein allocation during phase 2B of lactation, providing a maternal mechanism for differential allocation with fitness consequences. We believe collection of earlier phase 2A (0–100 days post partum) milk may yield important results given that differential investment in metatherians may be most apparent early in lactation, prior to any significant maternal investment, when a decision on termination of investment can be made with very little energetic loss to the mother. Interestingly, small mothers did not birth sons and better maternal condition was associated with raising sons. These data are in support of TWH and demonstrate a potential mechanism through which condition dependent and sex specific maternal investment may occur.     

Evaluation of reference lactation length in Chios dairy sheep

Definition and establishment of a fixed reference lactation length could provide useful tools for on-farm comparison of ewes and flock management as well as genetic evaluations for the breeding programme. The objectives of this study were to (i) evaluate different reference lactation lengths for the Chios dairy sheep and (ii) define the most suitable reference length for the breed. A total of 260 042 test-day milk records from 24 474 ewes in 130 flocks collected between 2003 and 2014 were used; 15 different lactation lengths were evaluated ranging from 120 to 260 days, defined at 10-day intervals as reference for the Chios sheep. The evaluation criteria included: (a) heritability and repeatability of milk yield in each reference lactation, (b) genetic correlation of reference lactation milk yield with actual lactation milk yield and yield at first test-day record and (c) correlated response in reference lactation milk yield from selection based on first test-day milk yield. The latter emulates genetic gains achieved for milk yield based on early lactation selection. Heritability and repeatability estimates of reference lactation milk yield and genetic correlation with actual lactation yield favoured long reference lactations (180 to 230 days). On the contrary, correlation with first test-day record milk yield was higher for short lactations (120 to 170 days). Moreover, selection on first test-day record milk yield would lead to a correlated response in reference yield in 220 days equal to 85% of the highest estimate achieved in the maximum reference length of 260 days (190 days when only considering first lactation milk yield). Based on the results of the present study, an overall reference lactation length for the Chios breed of 220 days post-lambing and a first lactation reference length of 190 days post-lambing are recommended.     

Genetic and phenotypic parameters of lactations longer than 305 days (extended lactations)

Test-day milk yield and somatic cell count data over extended lactation (lactation to 540-600 days) were analysed considering part lactations as different traits and fitting random regression (RR) models. Data on Australian Jersey and Holstein Friesian (HF) were used to demonstrate the shape of the lactation curve and data on HF were used for genetic study. Test-day data from about 100 000 cows that calved between 1998 and 2005 were used for this study. In all analyses, a sire model was used.When part lactations were considered as different traits, protein yield early in the lactation (e.g. first 2 months) had a genetic correlation of about 0.8 with protein yield produced after 300 days of lactation. Genetic correlations between lactation stages that are adjacent to each other were high (0.9 or more) within parity. Across parities, genetic correlations were high for both protein and milk yield if they are within the same stage of lactation. Phenotypic correlations were lower than genetic correlations. Heritability of milk-yield traits estimated from the RR model varied from 0.15 at the beginning of the lactation to as high as 0.37 by the 4th month of lactation. All genetic correlations between different days in milk were positive, with the highest correlations between adjacent days in milk and decreasing correlations with increasing time-span. The pattern of genetic correlations between milk yield in the second 300 days (301 to 600 days of lactation) do not markedly differ from the pattern in the first 300 days of lactation. The lowest estimated genetic correlation was 0.15 between milk yield on days 45 and 525 of lactation. The result from this study shows that progeny of bulls with high estimated breeding values for yield traits and those that produce at a relatively high level in the first few months are the most likely candidates for use in herds favouring extended lactations.     

Metabolic adaptations in goat mammary tissue during pregnancy and lactation

Metabolic adaptations of goat mammary tissue during pregnancy and lactation were monitored in serial biopsies of the tissue. Changes in the synthetic capacity of secretory cells were studied by combining measurements of enzyme activities with short-term culture of mammary explants to measure lactose, casein and total protein synthesis. By these criteria, the main phase of mammary differentiation began in late pregnancy and was essentially complete by Week 5 of lactation, coinciding with the achievement of peak milk yield. While milk yield declined after Week 5, the activities of key enzymes expressed per mg DNA and the rates of lactose and casein synthesis in mammary explants were maintained over a considerable period. The results suggest that changes in the synthetic capacity of epithelial cells may account for much of the rise in milk yield in early lactation, but are not responsible for the declining phase of milk production characteristic of lactation in ruminants.     

Choline and Choline Metabolite Patterns and Associations in Blood and Milk during Lactation in Dairy Cows

Milk and dairy products are an important source of choline, a nutrient essential for human health. Infant formula derived from bovine milk contains a number of metabolic forms of choline, all contribute to the growth and development of the newborn. At present, little is known about the factors that influence the concentrations of choline metabolites in milk. The objectives of this study were to characterize and then evaluate associations for choline and its metabolites in blood and milk through the first 37 weeks of lactation in the dairy cow. Milk and blood samples from twelve Holstein cows were collected in early, mid and late lactation and analyzed for acetylcholine, free choline, betaine, glycerophosphocholine, lysophosphatidylcholine, phosphatidylcholine, phosphocholine and sphingomyelin using hydrophilic interaction liquid chromatography-tandem mass spectrometry, and quantified using stable isotope-labeled internal standards. Total choline concentration in plasma, which was almost entirely phosphatidylcholine, increased 10-times from early to late lactation (1305 to 13,535 µmol/L). In milk, phosphocholine was the main metabolite in early lactation (492 µmol/L), which is a similar concentration to that found in human milk, however, phosphocholine concentration decreased exponentially through lactation to 43 µmol/L in late lactation. In contrast, phosphatidylcholine was the main metabolite in mid and late lactation (188 µmol/L and 659 µmol/L, respectively), with the increase through lactation positively correlated with phosphatidylcholine in plasma (R ² = 0.78). Unlike previously reported with human milk we found no correlation between plasma free choline concentration and milk choline metabolites. The changes in pattern of phosphocholine and phosphatidylcholine in milk through lactation observed in the bovine suggests that it is possible to manufacture infant formula that more closely matches these metabolites profile in human milk.     

The extracellular matrix locally regulates asynchronous concurrent lactation in tammar wallaby (Macropus eugenii)

Asynchronous concurrent lactation (ACL) is an extreme lactation strategy in macropod marsupials including the tammar wallaby, that may hold the key to understanding local control of mammary epithelial cell function. Marsupials have a short gestation and a long lactation consisting of three phases; P2A, P2B and P3, representing early, mid and late lactation respectively and characterised by profound changes in milk composition. A lactating tammar is able to concurrently produce phase 2A and 3 milk from adjacent glands in order to feed a young newborn and an older sibling at heel. Physiological effectors of ACL remain unknown and in this study the extracellular matrix (ECM) is investigated for its role in switching mammary phenotypes between phases of tammar wallaby lactation. Using the level of expression of the genes for the phase specific markers tELP, tWAP, and tLLP-B representing phases 2A, 2B and 3 respectively we show for the first time that tammar wallaby mammary epithelial cells (WallMECs) extracted from P2B acquire P3 phenotype when cultured on P3 ECM. Similarly P2A cells acquire P2B phenotype when cultured on P2B ECM. We further demonstrate that changes in phase phenotype correlate with phase-specific changes in ECM composition. This study shows that progressive changes in ECM composition in individual mammary glands provide a local regulatory mechanism for milk protein gene expression thereby enabling the mammary glands to lactate independently.     

Impact of indoor feeding at late lactation stage on body reserves recovery and reproductive performances of Baladi dairy goats fed on pastoral system

Adaptive capacities of Baladi goats facing situations of food restrictions and re-feeding during lactation have been studied. Three diets were tested: mountain natural rangelands during early lactation, agricultural pastures during mid-lactation and indoor stall feeding during late lactation. Body weight (BW), body condition (BCS) and plasma metabolites (NEFA) as well as milk production (milk yield, milk fat and milk protein contents) were measured in the different feeding situations. Effect of late lactation re-feeding on reproduction was assessed through kids’ birth weight and weight gain from day 0 till day 60 of age. Results showed that on rangelands, goats’ body condition degraded, with initially fat goats loosing more weight and being subject to a greater reserves mobilization than lean ones. These expressed an adaptive behaviour through increasing their daily feed intake and by an intense body reserves replenishment following their transfer to the agricultural pastures and indoor feeding. In late lactation, even goats kept on agricultural pastures showed a similar reconstitution, showing the high priority of body reserves replenishment during late lactation whatever the feeding level. The milk rebounds observed showed a good reactivity from the Baladi goat in response to a feed improvement either on agricultural pasture or indoor. At the end of lactation the milk yield decreased in all feeding situations proving the priority given by the Baladi goats to the body reserves replenishment over the milk production in order to ensure the next cycle. However, even if there was no difference in kids’ birth weights, kids from stall-fed dams had higher weight gains between 0 and 60 days of age when compared to those from underfed ones.     

Relationships between conception rate in Holstein heifers and cows and milk yield at various stages of lactation

We investigated the relationships between conception rates (CRs) at first service in Japanese Holstein heifers (i.e. animals that had not yet had their first calf) and cows and their test-day (TD) milk yields. Data included records of artificial insemination (AI) for heifers and cows that had calved for the first time between 2000 and 2008 and their TD milk yields at 6 through 305 days in milk (DIM) from first through third lactations. CR was defined as a binary trait for which first AI was a failure or success. A threshold-linear animal model was applied to estimate genetic correlations between CRs of heifers or cows and TD milk yield at various lactation stages. Two-trait genetic analyses were performed for every combination of CR and TD milk yield by using the Bayesian method with Gibbs sampling. The posterior means of the heritabilities of CR were 0.031 for heifers, 0.034 for first-lactation cows and 0.028 for second-lactation cows. Heritabilities for TD milk yield increased from 0.324 to 0.433 with increasing DIM but decreased slightly after 210 DIM during first lactation. These heritabilities from the second and third lactations were higher during late stages of lactation than during early stages. Posterior means of the genetic correlations between heifer CR and all TD yields were positive (range, 0.082 to 0.287), but those between CR of cows and milk yields during first or second lactation were negative (range, −0.121 to −0.250). Therefore, during every stage of lactation, selection in the direction of increasing milk yield may reduce CR in cows. The genetic relationships between CR and lactation curve shape were quite weak, because the genetic correlations between CR and TD milk yield were constant during the lactation period.     

Milk secretion in the rat: progressive changes in milk composition during lactation and weaning and the effect of diet.

1. Progressive changes in the composition of milk from rats has been studied from day 0 to 20 of lactation and for 3 days following separation of the dams and pups at day 20 post partum. 2. The changes in concentration of Na, K and lactose suggested that secretion both prepartum and following weaning occurred by a paracellular mechanism whereas a transcellular pathway existed during established lactation. 3. The concentration of total protein and casein increased gradually throughout lactation. In contrast, the concentration of serum albumin increased and transferrin decreased markedly during early lactation. The fat content of milk declined 3-fold within 5 days of birth but the concentration of Ca, Mg and inorganic P increased. The concentration of each of these milk constituents remained constant during established lactation. 4. Following weaning the pronounced decline in lactose, K and inorganic P was negatively correlated with an increase in all other milk constituents except fat. 5. Rats fed a low energy diet produced milk with a lower fat content but with an unaltered concentration of protein and carbohydrate. The growth rate of these litters was similar for the first 5 days of lactation when compared to litters from dams fed a high energy diet. The growth rate of litters thereafter and following weaning was greater for rats fed a high energy diet.     

Fatty acid mobilization and comparison to milk fatty acid content in northern elephant seals

A fundamental feature of the life history of true seals, bears and baleen whales is lactation while fasting. This study examined the mobilization of fatty acids from blubber and their subsequent partitioning into maternal metabolism and milk production in northern elephant seals (Mirounga angustirostris). The fatty acid composition of blubber and milk was measured in both early and late lactation. Proportions of fatty acids in milk and blubber were found to display a high degree of similarity both early and late in lactation. Seals mobilized an enormous amount of lipid (~66 kg in 17 days), but thermoregulatory fatty acids, those that remain fluid at low temperatures, were relatively conserved in the outer blubber layer. Despite the stratification, the pattern of mobilization of specific fatty acids conforms to biochemical predictions. Long chain (>20C) monounsaturated fatty acids (MUFAs) were the least mobilized from blubber and the only class of fatty acids that showed a proportional increase in milk in late lactation. Polyunsaturated fatty acids (PUFAs) and saturated fatty acids (SFAs) were more mobilized from the blubber, but neither proportion increased in milk at late lactation. These data suggest that of the long chain MUFA mobilized, the majority is directed to milk synthesis. The mother may preferentially use PUFA and SFA for her own metabolism, decreasing the availability for deposition into milk. The potential impacts of milk fatty acid delivery on pup diving development and thermoregulation are exciting avenues for exploration.     

Short- and long-term effects of high milking frequency during the first 21 days of lactation on production and reproductive performance in high-lactating cows

We examined short- and long-term effects of high milking frequency (HMF) for the first 21 days of lactation. The study included 122 Israeli Holstein cows – 32 pregnant heifers, 40 cows in second lactation and 50 cows in >second lactation. Heifers were paired according to predicted transmitting ability and cows according to energy-corrected milk (ECM) production, age, days in milk and expected calving date. Thin cows (body condition score <2.75) were not included. One cow from each pair was arbitrarily allocated to a control group milked three times daily (3× milking cows) and the counterpart to an experimental group milked six times daily for the first 21 days of lactation and then three times daily for the rest of the lactation (6× milking cows). During the first 21 days of lactation, 6× milking cows produced 9.3 kg more milk (26.5%) and 7.16 kg more ECM (19%) than the 3× milking cows. The higher milk production persisted throughout the entire lactation (305 days), as reflected by treatment×age interaction showing higher milk production for the first and second (7%) but not >second lactation cows relative to their control counterparts (−0.37%); ECM production was also higher in 6× milking first and second lactation (7.6% and 5%, respectively) but not for >second lactation cows. Furthermore, HMF had long-lasting effects, expressed as significantly higher milk production through the succeeding lactation in the previous first lactation cows (10%); a tendency toward significance in the second lactation cows relative to the controls (4.7%), but a deleterious effect on the >second lactation cows, reflected by lower milk production (−5.25%) than in controls; similar patterns were found for the ECM. For the entire 305 days of lactation, fat and protein yields were higher for first and second lactation cows, whereas protein yield for >second lactation cows was lower in the 6× milking v. control group. Given that HMF during the first 21 days of first or second lactation increases milk and ECM yields throughout the concurrent and successive lactation with no adverse effect on energy balance, mastitis, metabolic diseases or reproduction, it seems to be economically beneficial. However, caution should be paid for >second lactation cows due to absence of significant effect in the entire of the first HMF applied lactation and the deleterious effect in the succeeding lactation.     

Proteomic analysis of whey and casein proteins in early milk from the marsupial Trichosurus vulpecula,the common brushtail possum

Whey and casein proteins representing the first and second halves of the early lactation phase in the common brushtail possum (Trichosurus vulpecula) have been compared by two dimensional gel electrophoresis. Nine components of whey were differentially expressed during early lactation, including proteins identified as cathepsin B, clusterin, late lactation protein, lysozyme, ganglioside M2 activator and neutrophil gelatinase-associated lipocalin. A major novel protein, termed very early lactation protein (VELP), was identified in whey. Partial amino acid sequence data obtained from VELP did not appear to match any other reported protein sequence. VELP was shown to be an acidic glycoprotein of 20–30 kDa which exists as a homodimer. In the casein fraction, κ-casein appeared to be differentially post-translationally modified during early lactation and fragments of β-casein were relatively more abundant at the earlier lactation stage.     

Genetic correlations between production and disease traits during first lactation in Holstein cows

The aim of this study was to estimate genetic correlations between milk yield, somatic cell score (SCS), mastitis, and claw and leg disorders (CLDs) during first lactation in Holstein cows by using a threshold–linear random regression test-day model. We used daily records of milk, fat and protein yields; somatic cell count (SCC); and mastitis and CLD incidences from 46 771 first-lactation Holstein cows in Hokkaido, Japan, that calved between 2000 and 2009. A threshold animal model for binary records (mastitis and CLDs) and linear animal model for yield traits were applied in our multiple trait analysis. For both liabilities and yield traits, additive genetic effects were used as random regression on cubic Legendre polynomials of days on milk. The highest positive genetic correlations between yields and disease incidences (0.36 for milk and mastitis, 0.56 for fat and mastitis, 0.24 for protein and mastitis, 0.32 for milk and CLD, 0.44 for fat and CLD and 0.31 for protein and CLD) were estimated at about the time of peak milk yield (36 to 65 days in milk). Selection focused on early lactation yield may therefore increase the risk of mastitis and CLDs. The positive genetic correlations of SCS with mastitis or CLD incidence imply that selection to reduce SCS in the early stages of lactation would decrease the incidence of both mastitis and CLD.     

Histological and histochemical study on mammary gland of Damascus goats through stages of lactation

This research was carried out on seven Damascus goats, to study the relationship between milk production, during advancing lactation, and the changes in secretory mammary cells frequency and cellular activity. Biopsies were obtained from the mammary gland at the three stages of lactation, early, mid and late, for histological and histochemical studies. The histological structures of the mammary gland showed clear differences between lactation stages—being more developed in the early and the mid stages, compared to the late stage of lactation. The number of the alveolar secretory cells increased from the early to the mid stage of lactation by 12.9% and then was reduced at the late stage by 35.9% from that at the mid stage. The milk yield increased by 51.3% from the early to the mid stage, and then was reduced at the late stage by 71.4% from that of the mid stage. The total sectional areas of plate equal to 1.22 mm²/plate of the alveoli were the smallest during late lactation (0.36 mm²/plate) compared to that during the early and the mid stage of lactation (0.50 and 1.17 mm²/plate, respectively). Numerous loci of alkaline phosphatase enzyme (AP) were apparent on the outer surface of the alveolar secretory cells at the early and the mid stages of lactation—suggesting that this enzyme plays an important physiological role in the apical membrane of the alveolar epithelial cells during lactation. Dense protein staining of these cells as well as increased frequency of DNA expression denote great development and increased numbers of these cells at early and mid stages of lactation. This was accompanied by a high level of milk secretion reaching 939.3 ± 130 and 1421.4 ± 123.4 ml/head/day, respectively. In contrast, at the late stage of lactation, the size of alveoli was reduced and few alveoli showed weak AP activity, weak protein manifestation and the lowest frequency of DNA loci. This coincided with the reduction in milk yield (407 ml/head/day). It could be concluded that the stages of the lactation influence the cell number and activity of the mammary parenchyma.