Expression of parathyroid hormone-related protein (PTHrP) mRNA in mammary gland of periparturient cows

The expression of parathyroid hormone-related protein (PTHrP) mRNA was examined in mammary gland with or without lactation, and during periparturient period in a Holstein cow and a Jersey cow. In the lactating mammary gland, PTHrP was detected in alveolar epithelial cells and the lumen by immunohistochemical analysis. The relative expression levels of PTHrP mRNA in mammary gland from lactating cows were significantly higher than those from non-lactating cows (P<0.05). During periparturient period, relative PTHrP mRNA level was remarkably low before the parturition in a Jersey and a Holstein cow, however, both levels were gradually increased and reached a peak level at 5-6 weeks after the parturition. In addition, the peak level in a Jersey cow was approximately 3-fold higher than that in a Holstein cow. From these results, PTHrP was synthesized and secreted in alveolar epithelial cells in mammary gland and increased subsequently with the lactation, suggesting a possible mechanism for the regulation of local calcium homeostasis.     

Influence of different calcium contents in diets supplemented with anionic salts on bone metabolism in periparturient dairy cows

At the initiation of lactation, Ca homeostatic mechanisms have to react to a tremendous increase in demand for Ca. Mobilization of Ca from bone and increased absorption from the gastrointestinal tract are required to re-establish homeostasis. It has been shown that dietary anions play an important role in the prevention of milk fever by mobilizing Ca from bone and by increasing Ca absorption in the GI tract. The purpose of the present study was to investigate the influence of different Ca contents in diets supplemented with anionic salts on bone metabolism of dairy cows. Twenty-four holstein cows (housed inside, second to fourth lactation) without a milk fever history were divided into four groups (A, B, C, D). Each group was fed a different diet which was given from day 263 of gestation till the day of parturition. Group A and B received a low calcium diet (4 g/kg DM) whereas group C and D received a high Ca diet (8 g/kg DM). In addition group B and D received anionic salts. The DCAD was calculated with the formula: DCAD (mEq/kg DM)=(0.2 Ca2++0.16 Mg2++Na++K+)-(Cl-+0.6 S2-+0.65 P3-). Blood and urine samples were collected on days 256, 270 and 277 of gestation, on the day of parturition as well as the following 5 days and on days 9, 14 and 19 after parturition. Serum Ca, P, Mg, ICTP, OC, VITD, PTH and urinary pH were analysed. The bone resorption marker ICTP showed a significant increase after parturition in all the groups. On the contrary, the bone formation marker OC decreased after parturition in all the groups. The VITD concentrations in group D and the urinary pH in group B were significantly lower compared to the other groups (p<0.05). The Ca concentrations tended to be higher in group B around parturition than in all the other groups. No significant influence of the four different diets on all the other parameters could be shown. In conclusion, this data showed that the addition of anions and the different Ca contents had no significant influence on bone resorption and bone formation markers. This may be because of the fact that the dietary cation-anion balance was not low enough (DCAD-group A: 181 mEq/kg DM, group B: -48 mEq/kg DM, group C: 210 mEq/kg DM and group D: 28 mEq/kg DM) to induce a metabolic acidosis with all its positive effects on calcium metabolism.     

Genetics and physiology of leptin in periparturient dairy cows

In dairy cattle, the increase in milk yield has been accompanied by a more negative energy balance (EB) during early lactation and a decrease in fertility. As the hormone leptin is involved in regulation of nutritional status and reproductive function this hormone is an interesting protein to investigate during the periparturient period in dairy cattle. This study was performed to get insight into the function of leptin during the periparturient period and to perform an association study between polymorphisms in the bovine leptin gene and leptin receptor gene and fertility as well as production traits. Leptin concentrations in the periparturient cow undergo remarkable changes; leptin concentrations were high during late pregnancy and declined to a nadir at parturition. Genetic analysis of the leptin gene indicated that a combination of three polymorphisms located at a 135 bp region of the leptin promoter explained most of the variance in prepartum leptin concentrations. The two extreme genotype combinations could be used to investigate the function of leptin concentrations in pregnant cows. A polymorphism located on intron 2 of the leptin gene explained a significant part of the variation in milk yield. On the promoter region of the leptin gene an SNP was detected that was associated with first postpartum luteal activity (FPLA). This SNP could be a candidate marker for fertility in dairy cows. Another SNP on the leptin promoter was associated with energy balance and dry matter intake (DMI) where a higher dry matter intake occurred together with a higher energy balance. Two genotype combinations of the aforementioned three associated SNPs were defined which had a good milk yield together with a good energy balance and fertility. Calculations of an economical value per trait have to validate if one of these genotype combinations would be a possible candidate to be used in selection.     

Plasma parameters related to energy and lipid metabolism in periparturient Modenese and Italian Friesian cows

The aim of this work was to compare energy and lipid metabolism during the peripartum period between Modenese (MO) and Italian Friesian (IF) cows. The study was carried out on 33 pluriparous pregnant cows, 19 IF and 14 MO, reared together in the same herd and kept under equal conditions of management and nutrition. Blood was sampled from jugular vein starting 4 week before expected calving date until 4 week post‐calving, once weekly. Plasma was analysed for glucose, NEFA, BHBA, triglycerides and cholesterol concentrations. Body condition score (BCS) was assessed weekly after blood sampling. Data from antepartum (a.p.) and post‐partum (p.p.) periods were separately analysed as repeated measures by a linear mixed models with the effect of breed, time and their interaction as main factors and random cow within breed. The energy status differed between the two breeds during the peripartum period. We observed higher BCS a.p. and p.p. and lower BCS variations p.p. in MO compared to IF group. Modenese cows showed lower glucose and cholesterol concentrations (p < 0.001), but higher NEFA values, NEFA to cholesterol and NEFA to albumin ratios (p < 0.001) during a.p.; on the contrary, IF cows had higher (p < 0.05) cholesterol, NEFA, BHBA levels and NEFA to albumin ratio than MO ones during p.p. The differences observed between the two breeds suggest how MO cows are subjected to lipid mobilization during late gestation; on the contrary, IF cows are predisposed to mobilize their lipid reserves at the beginning of lactation to support high production. The results indicate a diverse ability to cope with metabolic stress and suggest the hypothesis that the differences in concentrations of plasma parameters and their variation amplitude around the calving period might depend on the different genetic merit for milk production between the two breeds.     

Plasma concentrations of parathyroid hormone-related protein in dogs with potential disorders of calcium metabolism

The plasma concentrations of total calcium, ionised calcium, albumin, parathyroid hormone and parathyroid hormone-related protein (PTHrp) were measured in 25 dogs with lymphoma, nine dogs with primary hyperparathyroidism and seven dogs with adenocarcinoma of the apocrine gland of the anal sac. Plasma total calcium, ionised calcium, albumin and parathyroid hormone-related protein were measured in 18 clinically normal control dogs. The concentration of PTHrp was high in 12 of the 14 dogs that were hypercalcaemic because of an underlying malignancy but was within the reference range in all the control dogs, in the 17 normocalcaemic dogs with lymphoma and in the seven dogs which were hypercalcaemic because of a parathyroid adenoma.     

Metabolic stress and inflammatory response in high-yielding, periparturient dairy cows

Increased disease rates are commonly reported among high-yielding dairy cows in the transition period, extending from 3 weeks before to 3 weeks after calving, and characterized by the occurrence of an inflammatory response in terms of both positive and negative acute phase proteins (APP+ and APP-). To determine the above inflammatory response, the authors had developed the Liver Functionality Index (LFI), which defines the above condition on the basis of some APP- responses (albumin, cholesterol sensu stricto+bilirubin) during the first month of lactation. In this respect, low LFI values are associated to a high inflammatory response and vice versa. The relationship between LFI and inflammatory cytokine response was investigated from day -28 to day +28 with respect to calving in 12 periparturient dairy cows showing the six highest and six lowest LFI values within a cohort of 54 high-yielding dairy cows. The hypothesis being tested was that LFI and APP- on the whole could be used as readout of successful vs. non-successful adaptation to the transition period, with a strong association to disease occurrence. In fact, low LFI cows experienced many more disease cases (13 vs. 3 in high LFI Group) and related drug treatments till day +28. Interleukin-6 (IL-6) serum concentrations were always higher in low LFI cows (P<0.05 on day +28). The greater IL-6 levels were correlated with higher ceruloplasmin (APP+) and lower lysozyme serum concentrations (P<0.05 and <0.1, respectively). This latter finding was correlated with a clear role in vitro of lysozyme in a dose-dependent modulation of the inflammatory response of swine intestinal epithelial cells and bovine peripheral blood mononuclear cells. Hematological examinations showed no significant differences between the two groups under study. On the whole, our results indicate that LFI and LFI-related parameters could be used to identify cows at risk in the transition period toward an improved farm management. Also, our study indicates that disease cases in periparturient, high-yielding dairy cows are correlated with signs of accentuated IL-6 response and other markers of inflammatory phenomena. These likely start in the late lactation period or around dry-off, as suggested by our prepartal data, and proceed at much greater levels after calving.     

Serum osteocalcin in dairy cows: age-related changes and periparturient variation

We evaluated age-related changes in serum osteocalcin concentrations in non-periparturient cows and variations in serum osteocalcin concentration in periparturient primiparous and multiparous cows. The serum osteocalcin levels were evaluated in 144 non-periparturient Holstein dairy cows aged 11 days to 10 years; these levels were the highest in the youngest cows, appeared to steadily decrease with age until the time of the first calving, and were subsequently maintained at low levels. Between 14 days before calving and 21 days after calving, the serum osteocalcin levels were significantly higher in the primiparous cows than in the multiparous cows. A comparison between age-matched non-periparturient and periparturient cows showed that serum osteocalcin levels were significantly lowered during late gestation in both primiparous and multiparous cows. These results suggest that serum osteocalcin measurement might be useful for the detection of mineral imbalances at the time of parturition in cows.     

Changes in oxidative status in periparturient dairy cows in hot conditions

We investigated the changes in the plasma concentrations of oxidative stress markers in periparturient dairy cows in hot conditions. This study was performed on 10 cows from day ?5 to day 10 after parturition. The experimental duration was divided into three periods: period 1 (5 days before parturition); period 2 (5 days after parturition); and period 3 (from day 6 to day 10 after parturition). The average rectal temperature in the cows during the experimental period was more than 39°C. Plasma aspartate aminotransferase activities in periods 2 and 3 were higher than that in period 1 (P < 0.05). Blood urea nitrogen concentration in period 3 was lower than those in periods 1 and 2 (P < 0.05). The total ascorbic acid concentration in the plasma in period 2 was greater than those in periods 1 and 3 (P < 0.05). Thiobarbituric acid reactive substances (TBARS) concentrations in periods 2 and 3 were greater than that in period 1 (P < 0.05). TBARS concentration was positively correlated with aspartate aminotransferase activity (P < 0.05). These results suggested that oxidative stress increased in cows after parturition under hot climatic conditions.     

A proposed modulatory role of the endocannabinoid system on adipose tissue metabolism and appetite in periparturient dairy cows

To sustain the nutrient demands of rapid fetal growth, parturition, and milk synthesis, periparturient dairy cows mobilize adipose tissue fatty acid stores through lipolysis. This process induces an inflammatory response within AT that is resolved as lactation progresses; however, excessive and protracted lipolysis compounds the risk for metabolic and inflammatory diseases. The suppression of lipolytic action and inflammation, along with amplification of adipogenesis and lipogenesis, serve as prospective therapeutic targets for improving the health of periparturient dairy cows. Generally, the activation of cannabinoid receptors by endocannabinoids enhances adipogenesis and lipogenesis, suppresses lipolysis, and increases appetite in mammals. These biological effects of activating the endocannabinoid system open the possibility of harnessing the endocannabinoid system through nutritional intervention in dairy herds as a potential tool to improve dairy cows' health, although much is still to be revealed in this context. This review summarizes the current knowledge surrounding the components of the endocannabinoid system, elaborates on the metabolic effects of its activation, and explores the potential to modulate its activity in periparturient dairy cows.     

Characterization of ruminal dynamics in Holstein dairy cows during the periparturient period

We used four pregnant Holstein cows to delineate ruminal adaptations as cows transitioned from one lactation to the next. Cows were fed typical diets through far-off and close-up dry periods and lactation. We measured ruminal characteristics on day 72 (late lactation), 51 (far-off dry), 23 and 9 (close-up dry) prepartum and on days 6, 20, 34, 48, 62, 76 and 90 postpartum (early lactation). Measurements included: ruminal fill (weight of actual contents), ruminal capacity (volume of rumen when fully filled), digestibilities and ruminal passage rates. Ruminal capacity tended to increase linearly during early lactation but was stable during dry and transition periods. Both total and liquid fill decreased linearly during the dry period, increased across parturition, and increased linearly through early lactation. Dry matter fill decreased as cows were fed the close-up diet at day 23 prepartum then increased near parturition and continued to increase across early lactation. Solid passage rate was greatest when cows were fed the close-up diet, and decreased throughout the transition period. In lactation, solid passage rate responded quadratically with peak at day 48 followed by decreases through day 90 postpartum. Liquid passage increased linearly across the transition period. Total tract organic matter digestibilities increased linearly over the dry period with significant increases prior to or immediately after parturition, then they remained relatively stable over early lactation until they increased at day 90. Fibre digestibilities demonstrated quadratic responses over early lactation, being higher on day 6 and day 90 than at other times. Starch digestibilities decreased linearly across both the dry and transition periods with decreases in lactation until day 62 followed by increases until day 90. High producing lactating dairy cows go through a multitude of ruminal adaptations, in terms of digestion, passage, capacity and fill, as they transition from one lactation to the next.     

Fecal shedding of coliform bacteria during the periparturient period in dairy cows

OBJECTIVE: To determine whether numbers of coliform bacteria in feces of dairy cattle changed during the periparturient period and whether fluctuations were associated with changes in dry-matter intake. ANIMALS: 12 healthy Holstein cows. PROCEDURE: Fecal samples were collected on a semi-regular basis (i.e., 3 to 7 times/wk) beginning 4 to 6 weeks before the anticipated parturition date and continuing through the third day (5 cows) or second week (7 cows) after parturition, and total numbers of fecal coliform bacteria were determined. Daily feed intake of 7 cows was monitored. RESULTS: For 11 cows, fecal coliform bacterial counts between 34 and 25 days prior to parturition were low and relatively constant (< 102 change in number of bacteria). Coliform bacteria were not detected in 4 to 8% of fecal samples from 10 cows. All cows had a 10(4) to 10(7) increase in number of colony forming units/g of feces near the time of parturition. Number of fecal coliform bacteria peaked within 7 days of parturition in 9 cows and within 12 days of parturition in 3. Number of fecal coliform bacteria was not correlated with feed intake. CONCLUSIONS AND CLINICAL RELEVANCE: Cows may have large increases in fecal coliform bacteria count during the periparturient period; however, periparturient cows do not continually shed high numbers of coliform bacteria, and coliform bacteria may not always be detectable by conventional culture methods. Changes in fecal coliform bacteria count did not correlate with changes in dry-matter intake.     

Changes of vitamin E status of periparturient dairy cows and newborn calves

Eleven primiparous Holstein Friesian and their crossbred calves (F1, Japanese Black cattle × Holstein Friesian) and 10 multiparous Holstein Friesian and their Holstein Friesian calves were used to evaluate vitamin E status in periparturient period. Plasma α‐tocopherol (α‐toc) concentrations of the multiparous cows were significantly higher than those of the primiparous cows from 60 days before expected calving to 90 days of lactation (P < 0.05). The multiparous cows had a further decrease in the concentrations of α‐toc and total lipid in plasma to the calving than the primiparous cows. Colostrum α‐toc concentrations in multiparous cows were significantly higher than those of the primiparous cows (P < 0.05). Plasma α‐toc concentrations of calves borne by the multiparous cows were significantly higher than those of the primiparous cows at 5 days of age (P < 0.05). Plasma α‐toc concentrations of calves were highest at 5 and 15 days of age in the calves borne by the multiparous and primiparous cows, respectively, and decreased thereafter till 90 days of age. The higher vitamin E status of multiparous cows over primiparous cows might have reflected nutritional composition in the rations. Their calves afforded higher plasma α‐toc levels after birth because of more α‐toc transfer via placenta and more α‐toc secretion in the colostrums thereafter. Plasma α‐toc concentrations of the calves might have decreased as the calves became dependent upon the solid feed of low vitamin E content.     

Plasma lipoproteins and fatty liver in dairy cows

Dairy cows were classified on the basis of a histological study after a hepatic biopsy conducted in the second week post partum (mild fatty liver, moderate fatty liver, severe fatty liver). Plasma lipoproteins were separated into various density classes by repeated ultracentrifugation. The results indicate that the beginning of lactation is associated with a low concentration of the 1.006 to 1.063 g ml-1 lipoprotein fraction. The lowest concentrations occurred in cows with severe steatosis or during the evolution of moderate steatosis.     

The effects of periparturient events on breeding performance of dairy cows

Many periparturient events affect breeding performance in dairy cows. These events include disorders, high early lactation milk yield, other cow factors, and calf factors. The effects range in severity from "annoying" to "devastating."     

Relationship between parity and mineral status in dairy cows during the periparturient period

Data from 65 cows were collected to clarify the relationship between parity and mineral status in Holstein cows during the periparturient period. Milk fever occurred in a second‐ and a fourth‐lactation cow at parturition. Bodyweight of cows decreased immediately after parturition, and the decreased weights of cows from 1 to 6 days after parturition ranged from 11 to 27 kg. Milk yield of multiparous cows was higher than that of primiparous cows, but Ca and P concentrations in colostrum were not affected by the parity. Plasma glucose of primiparous cows was higher than that of multiparous cows, and plasma nonesterified fatty acids of fourth‐and‐more‐lactation cows at parturition was highest. Plasma Ca concentration of cows at parturition decreased with advancing parity, and plasma inorganic P concentration of third‐and‐more‐lactation cows was lowest at parturition. Plasma parathyroid hormone of fourth‐and‐more‐lactation cows at parturition was highest, but plasma hydroxyproline and alkaline phosphatase was lowest. These results suggest that the large transfer of Ca and P to colostrum is a factor in the development of milk fever in third‐and‐more‐lactation cows.     

An immunogenomics approach to understanding periparturient immunosuppression and mastitis susceptibility in dairy cows

Studies comparing in vivo and in vitro functional capacities of leukocytes from non-parturient and periparturient dairy cows have provided substantial evidence that systemic and local mammary immune defenses are deficient around parturition. This evidence has lead to the reasonable hypothesis that immune deficiency underlies the heightened mastitis susceptibility of periparturient cows. Nutrition and vaccine studies substantiate this hypothesis, showing that dietary antioxidant supplementation and rigorous immunization regimes can bolster innate and humoral immunity to the point that mastitis severity and time for return to normal milk production are reduced. However, completely effective resolution of this significant production disease has not been achieved because so little is understood about its complex etiology. In particular, we possess almost no knowledge of how or why immune cells responding to parturient physiology end up with deficient functional capacities. Fluctuations in reproductive steroid hormones and chronic shifts in neuroendocrine hormones with roles in nutrient partitioning and appetite control may affect the expression of critical leukocyte genes in periparturient dairy cows. A thorough understanding of leukocyte biology during periparturition would seem a critical goal for future development of effective mastitis prevention strategies. Recently, our group has begun to use cDNA microarray technology to explore bovine leukocyte RNA for global gene expression changes occurring around parturition. We are working within the context of a hypothesis that the physiology of parturition negatively affects expression of critical genes in blood leukocytes. In the current study we initiated hypothesis testing using leukocyte RNA from a high producing Holstein cow collected at 14 days prepartum and 6 hours postpartum to interrogate a cDNA microarray spotted with > 700 cDNAs representing unique bovine leukocyte genes. This analysis revealed 18 genes with > or = 1.2-fold higher expression 14 days prepartum than 6 hours postpartum. BLASTN analysis of these genes revealed only one that can be considered a classical immune response gene. All other repressed genes were either unknown or putatively identified as encoding key proteins involved in normal growth and metabolism of cells. Given the critical roles of these repressed genes in normal cell functions, we may have identified good candidates to pursue with respect to periparturient immunosuppression and mastitis susceptibility.