Hierarchy of factors affecting behavioural signs used for oestrus detection of Holstein and Normande dairy cows in a seasonal calving system

As oestrous expression of dairy cows has decreased over the last decades oestrus detection has become more difficult. The objective of this study is to identify the main factors that affect oestrus detection in seasonal calving dairy cows, and to establish their relative importance. In each of 5 years 36 Normande and 36 Holstein cows were assigned to a Low or High winter-feeding level group. Half of each group was then assigned to a Low or High pasture-feeding group. The Low-Low strategy resulted in the lowest milk yield and the greatest body condition (BC) loss from calving to nadir BC score (6302 kg; -0.98 unit). The High-High strategy had the converse effect (7549 kg; -0.75 units). Low-High and High-Low strategies had intermediate values. The Normande cows had lower milk yield and BC loss than Holstein cows (6153 kg versus 7620 kg; -0.82 unit versus -1.20 unit). A database of 415 observed spontaneous oestruses was created. Oestruses were classified according to detection signs: (1) standing to be mounted, (2) mounting without standing, (3) other signs without standing or mounting (slight signs). Presence of another cow in oestrus, access to pasture, Normande breed and Low-Low strategy increased standing detection. In the Normande breed, 97% of oestruses were detected by standing while combining the presence of a herdmate in oestrus and access to pasture with a milk production of less than 6550 kg. Holstein cows had a higher frequency of slight signs oestruses than Normande ones, which was associated with a decreased subsequent calving rate (P<0.05). In multiparous Holstein cows, the odds of slight signs detection was multiplied by 7.8 for the High-High group in comparison with the Low-Low group (P<0.05). In our study milk yield had an effect on oestrus detection which was not explained by BC loss. As High-High cows produced more milk than others, we logically found that an increase in milk yield increased slight signs detection. Conversely, as they lost less BC than others, BC loss improved the chance of standing or mounting detection. These two results show that an increase in milk yield may reduce oestrous behaviour even if BC loss is moderate. Oestrus detection is crucial in seasonal compact calving systems. High phenotypic milk yields appear unsuitable with such systems in regard to depressed oestrous behaviour.     

Does supplementing dairy cows with β-carotene during the dry period affect postpartum ovarian activity, progesterone, and cervical and uterine involution?

β-carotene is the main natural precursor of vitamin A and plays an important role in reproductive efficiency and immune function in dairy cows. The objective of this study was to investigate whether a supplement of β-carotene given during the dry period is able to 1) increase blood concentrations of β-carotene postpartum, 2) improve ovarian function and progesterone production, and 3) enhance uterine involution and uterine health. This study was conducted using 40 Holstein cows. On the day of drying-off, cows were allocated to one of two dietary treatments: control diet (C, n = 20) or control diet plus 1g/d β-carotene (BC, n = 20). The β-carotene supplement was given individually to the cows until calving. Blood samples were obtained regularly before and after calving from the cows to measure the concentrations of β-carotene. The diameters of the cervix and uterine horns were measured regularly using ultrasonography. Endometrial cytology samples were acquired from the cervix and uterus to determine uterine health. Milk samples were obtained three times per week for progesterone assay. Additional blood samples were taken on the day of calving, 7 and 21 days postpartum to determine the plasma concentrations of amino acids. Blood concentrations of β-carotene were not different before the start of the experiment (C, 3.03 ± 0.22 mg/L vs BC, 3.12 ± 0.22 mg/L, P > 0.05). Blood concentrations of β-carotene in the BC group peaked (7.45 ± 0.24 mg/L) 1 month after drying-off while the concentrations in the C group remained constant. β-carotene concentrations then decreased in both groups. The difference in blood concentrations of β-carotene between groups became significant 2 weeks after the start of the supplement until 2 weeks postpartum. There was no significant difference in the interval from calving to ovulation between groups (C, 27.8 ± 3.46 d vs BC, 35.8 ± 3.55 d, P > 0.05). The dietary supplement of β-carotene during the dry period had no effect on ovarian activity, progesterone production, cervix and uterine horn diameters. Plasma concentrations of hydroxyproline in the BC group were higher than in the C group on day 21 postpartum (BC, 20.8 ± 1.33 μmol/L vs C, 15.0 ± 1.33 μmol/L; P < 0.01). On day 28 postpartum the percentage of neutrophils in the BC group was lower than in the C group (cervical smear; C, 21.0 ± 3.22% vs BC, 9.7 ± 3.14%, P < 0.05 and uterine smear; C, 32.0 ± 3.86% vs BC, 20.9 ± 3.76%, P < 0.05). In the present experiment a dietary supplement of β-carotene had no effect on ovarian activity. However, due to effects of β-carotene on hydroxyproline profiles and their potential relationship with uterine function we speculate that uterine involution may have been more complete and that uterine inflammation may have been reduced in cows which received the β-carotene compared to controls.     

Influence of negative energy balance on cyclicity and fertility in the high producing dairy cow

The peripartum period is of critical importance to subsequent health and fertility. Most cows enter a state of negative energy balance (NEB) associated with many metabolic changes which have carry over effects on the resumption and normality of estrous cyclicity and the success of subsequent inseminations. A dataset on 500 lactations explored the relationships between metabolic traits measured before and after calving with fertility. Stepwise multiple regression analysis showed that longer calving to conception intervals were associated with altered profiles of IGF-I, urea and body condition score. These relationships between metabolic profiles and fertility differed between first lactation cows (which are still growing but produce less milk) and mature animals. Early postpartum the liver undergoes extensive biochemical and morphological modifications to adapt to NEB, the uterus is extensively remodeled and must clear bacterial infections, and the ovary must resume ovulatory cycles. RNA isolated from liver and uterine tissues harvested 2 weeks postpartum from cows in mild (MNEB) and severe (SNEB) energy balance was used to screen the Affymetrix 23K bovine microarray. In liver, SNEB resulted in differential expression of key genes involved in lipid catabolism, gluconeogenesis, and the synthesis and stability of IGF-I. This was accompanied by reduced systemic concentrations of IGF-I which is likely to impact on ovarian function and early embryo development. Within endometrium, cows in SNEB showed histological evidence for higher levels of inflammation and the microarray analysis identified groups of differentially expressed genes involved in tissue remodeling and immune response. This may delay uterine repair after calving, likely contributing to the observed reduction in fertility.     

Evaluation of cytokine expression by blood monocytes of lactating Holstein cows with or without postpartum uterine disease

Whereas neutrophils are the main phagocytic leukocytes, monocytes and macrophages are actively involved in immunomodulation after infection. Recent studies have demonstrated that neutrophil function is impaired by the state of negative energy balance around parturition, and that cows that develop uterine disease have a greater degree of negative energy balance than healthy cows. The objectives of this study were to compare monocyte gene expression and protein secretion of selected cytokines from calving to 42 d after calving in Holstein cows that did or did not develop uterine disease. Real time quantitative RT-PCR (Tumor necrosis factor-α (TNFα), Interleukin (IL)-1β, IL-6, IL-8 and IL-10) and ELISA (TNFα, IL-1β and IL-8) were used to evaluate cytokine response following in vitro stimulation of blood-derived monocytes with irradiated E. coli. Relative to unstimulated cells, E. coli-stimulated monocytes from cows with metritis had lower gene expression of key pro-inflammatory cytokines than healthy cows from calving to 14 d after calving (TNFα at 0, 7, and 14 d after calving, IL-1β and IL-6 at 7 and 14 d after calving; P < 0.05). There were no significant differences between groups for expression of IL-8 or the anti-inflammatory cytokine IL-10. This was due, in part, to higher gene expression in unstimulated monocytes (TNFα, IL-1β, IL-6 and IL-10) in early lactation from cows with metritis. Expression of mRNA in stimulated cells (relative to housekeeping genes) was lower for TNFα (7 and 14 d postpartum) and for IL-10 (7 and 14 d postpartum) in cows with metritis. Concentration of TNFα was lower in the culture medium of E. coli-stimulated monocytes from cows with metritis than healthy cows at calving and 7 and 21 d after calving (P < 0.05). Circulating cytokine concentrations were not different between groups for IL-8 and were below the limits of detection for TNFα and IL-1β. Cytokine gene expression and production were similar between healthy cows and cows that developed endometritis, diagnosed cytologically at 42 d after calving. We concluded that altered levels of expression and production of pro-inflammatory cytokines postpartum could contribute to impaired inflammatory response and predispose cows to development of metritis.     

Dairy cows' reproductive response to feeding level differs according to the reproductive stage and the breed

Reproductive performance has decreased over the last decades in many dairy systems. This study aimed at comparing the effects of a high- and a low-feeding level on reproductive stages (cyclicity, oestrus and fertility) of Holstein and Normande cows in a grass-based winter compact calving system. High-fed cows received a total mixed ration composed of 55% maize silage, 15% dehydrated alfalfa pellets and 30% concentrate in winter and 4 kg/day concentrate supply at grazing. Low-fed cows only received 50% grass silage and 50% haylage in winter and no concentrate at grazing. Low-fed cows produced less milk over 44 weeks, but lost more body condition (BC) than high-fed ones (5207 v. 7457 kg, -1.28 v. -0.96 unit, P < 0.001). Normande cows produced less milk and lost less BC than Holstein ones (5596 v. 7068 kg, -0.89 v. -1.36 unit, P < 0.001). Post-partum ovarian activity was little affected by the feeding level. In both breeds, ovulation detection rate was higher in low-fed cows (74% v. 59%, P < 0.001) and detection relied more frequently on standing behaviour (67% v. 55%, P = 0.022). In both breeds, the recalving rate following the first and second inseminations was not significantly affected by the feeding level, although more non-fertilisations or early embryo mortalities occurred in low-fed cows (43% v. 26%, P = 0.004). For the Holstein breed, this was clearly explained by more late embryo mortalities in high-fed cows (30% v. 9%, P = 0.004). Finally, pregnancy rate by the end of the 13-week breeding period was similar between feeding groups. Conversely, the Normande dual-purpose cows had a higher pregnancy rate by the end of the breeding period than the Holstein dairy cows (72% v. 54%, P = 0.007), owing to a better ovarian activity (79% v. 54% normal, P < 0.001) and a higher recalving rate following insemination (53% v. 37%, P = 0.007). In conclusion, this study demonstrates that feeding levels with converse effects on milk yield and BC score also have converse effects on reproductive stages and lead to quite similar final reproductive performance in compact calving systems. Normande cows benefit from an overall better reproductive performance, but do not suit high feeding levels for very compact calving systems owing to depressed oestrous behaviour.     

Lymphocyte functions in dairy cows in hot environment

This study was carried out to ascertain the effects of intense high environmental temperatures (HET) on lymphocyte functions in periparturient dairy cows. The study was undertaken from the beginning of March through the end of July 2003 in a commercial dairy unit located approximately 40 km north of Rome. Thirty-four Holstein cows were utilised in the study. Twenty-two of these cows gave birth in spring (SP cows), from 28 March to 30 April. The remaining 12 cows gave birth in summer (SU cows), between 15 June and 2 July. The two groups of cows were balanced for parity and were fed the same rations. Blood samples were taken 4, 3, 2 and 1 week before calving, and 1, 2 and 4 weeks after calving, in order to evaluate peripheral blood mononuclear cell (PBMC) function in vitro, and to determine plasma cortisol concentrations. After isolation, the PBMC were stimulated with mitogens and their response in terms of DNA synthesis and IgM secretion was measured. During spring, either the day (9–20 h) or the night (21–8 h) temperature humidity index (THI) was below the upper critical THI (72) established for dairy cows. During summer, the mean daily THI values were 79.5±2.9 during the day and 70.1±4.7 during the night. Furthermore, during summer, three heat waves (a period of at least 3 consecutive days during which there were less than 10 recovery hours) occurred. Recovery hours were intended hours with a THI below 72. The first heat wave lasted 5 days, the second 6 days, and the third 15 days. Compared to the SP cows, over the entire periparturient period the extent of DNA synthesis and IgM secretion levels were lower (P ranging from <0.01 to 0.0001) and higher (P<0.01) respectively, in the SU cows. Before calving, the SU cows also presented higher (P<0.01) concentrations of plasma cortisol compared to the SP cows. This study indicates that the effects of HET on the immune response depend on the specific immune function under consideration, and that neuroendocrinal changes due to HET may play a role in the perturbation of immune functions.     

Changes in some Blood Micronutrients, Leukocytes and Neutrophil Expression of Adhesion Molecules in Periparturient Dairy Cows

Dairy cows are highly susceptible to infectious diseases, like mastitis, during the period around calving. Although factors contributing to increased susceptibility to infection have not been fully elucidated, impaired neutrophil recruitment to the site of infection and changes in the concentrations of some micronutrients related with the function of the immune defence has been implicated. Most of the current information is based on studies outside the Nordic countries where the conditions for dairy cows are different. Therefore, the aim of the study was to evaluate changes in blood concentrations of the vitamins A and E, the minerals calcium (Ca), phosphorous (P), and magnesium (Mg), the electrolytes potassium (K) and sodium (Na) and the trace elements selenium (Se), copper (Cu) and zinc (Zn), as well as changes in total and differential white blood cell counts (WBC) and expression of the adhesion molecules CD62L and CD18 on blood neutrophils in Swedish dairy cows during the period around calving. Blood samples were taken from 10 cows one month before expected calving, at calving and one month after calving. The results were mainly in line with reports from other countries. The concentrations of vitamins A and E, and of Zn, Ca and P decreased significantly at calving, while Se, Cu, and Na increased. Leukocytosis was detected at calving, mainly explained by neutrophilia, but also by monocytosis. The numbers of lymphocytes tended to decrease at the same time. The mean fluorescent intensity (MFI) of CD62L and CD18 molecules on blood neutrophils remained constant over time. The proportion of CD62L⁺ neutrophils decreased significantly at calving. The animals were fed according to, or above, their requirements. Therefore, changes in blood levels of vitamins, minerals and trace elements were mainly in response to colostrum formation, changes in dry matter intake, and ruminal metabolism around calving. Decreased levels of vitamins A and E, and of Zn at calving might have negative implications for the functions of the immune defence. The lower proportion of CD62L+ neutrophils at calving may result in less migration of blood neutrophils into the tissues, and might contribute to the increased susceptibility to infections at this time.     

Chemotactic activity of cotyledons for mononuclear leukocytes related to occurrence of retained placenta in dexamethasone induced parturition in cattle

Induction of parturition with glucocorticosteroids in cattle is used for research purposes, in diseased or injured pregnant cows, and as a management tool to time parturition. A negative side effect of induction of parturition with glucocorticosteroids is the high incidence of retained placenta that occurs after these calvings. Reaction of the maternal immune system against the ‘foreign’ foetal membranes contributes to the breakdown of the foetal-maternal attachment. Several studies indicate that failure of this immune assisted detachment increases the occurrence of retained placenta. We hypothesized that retained placenta occurring after induction of parturition with glucocorticosteroids is caused by failure of immune assisted detachment of the foetal membranes. The chemotactic activity of cotyledons for mononuclear leukocytes was used as a parameter to see whether immune assisted detachment of the foetal membranes had occurred. Cotyledons were collected from spontaneously calving non-retained placenta cows and from dexamethasone induced non-retained placenta and retained placenta cows. The study showed that the chemotactic activity of cotyledons for mononuclear leukocytes was lower (P < 0.001) in cotyledons obtained from retained placenta cows in which parturition was induced with dexamethasone compared to the chemotactic activity of cotyledons obtained from spontaneously calving non-retained placenta cows, whereas the chemotactic activity of cotyledons obtained from induced non-retained placenta cows was not lower (P = 0.10) than the chemotactic activity of cotyledons obtained from spontaneously calving non-retained placenta cows. We concluded that induction of parturition with dexamethasone causes a failure of immune assisted detachment of the foetal membranes and the accompanying release of chemotactic factors. As a result, the chemotactic activity of cotyledons for mononuclear leukocytes is lower in induced retained placenta cows than in cotyledons from non-retained placenta cows in which successful immune assisted detachment of the foetal membranes occurs.     

5-Azacytidine treatment of a murine cytotoxic T cell line alters interferon-gamma gene induction by interleukin 2

Genetic susceptibility to multiple sclerosis (MS) in Caucasians was previously shown to be correlated to the presence of given alleles at the HLA-DR and Gm loci. We now demonstrate that the humoral immune response in MS central nervous system (CNS) is modulated by both loci: the levels of IgG1 subclass and IgG1 allotypes in cerebrospinal fluid of MS patients depend on both their Gm genotype and their HLA-DR2 or HLA-DR7 phenotype. That HLA-DR molecules may either participate in a preferential recruitment of IgG1 allotype-producing B cells in MS CNS or act after such a selective homing is discussed. These results demonstrate that both HLA and Gm loci are synergistically involved in the modulation of the humoral immune response.     

Serum luteinizing hormone, 13,14-dihydro-15-keto-prostaglandin F2alpha and cortisol profiles during postpartum anestrus in Brahman and Angus cows

Pluriparous suckled Brahman and Angus cows were utilized to evaluate the effect of breed, day after calving and endogenous opioid peptides (EOP) on hormonal profiles during postpartum anestrus. On Days 17 and 34 after calving, blood samples with and without heparin were collected at 15- and 30-min intervals, respectively, for a 7-h period via jugular cannula. Two hours after the start of blood sampling, cows of each breed were administered either 1 mg/kg iv naloxone or saline. Three hours later, all animals received 10 ng/kg iv GnRH. On Day 34 after calving cows received 0.2 IU/kg iv ACTH. Mean LH, basal LH and area under the LH curve increased (P < 0.01) from Day 17 to Day 34 after calving. Height of LH pulses increased (P < 0.05) by Day 34 after calving. Brahman cows had higher (P < 0.05) mean LH, basal LH, LH pulse frequency and area under the LH curve than Angus cows. Naloxone increased postchallenge area under the LH curve in treated cows above that of control cows (P < 0.06). Naloxone also increased the postchallenge area under the LH curve above that of the prechallenge level (P < 0.01). No breed differences in the response to the naloxone challenge were observed. The LH response to naloxone challenge occurred earlier on Day 34 than on Day 17 after calving but the amount of LH released was similar between days. The GnRH-induced LH release was greater in Brahman than in Angus cows (P < 0.04). Mean cortisol concentrations and area under the cortisol curve decreased (P < 0.05) between Day 17 and Day 34 after calving. Mean cortisol concentrations and area under the cortisol curve were lower (P < 0.01) in Brahman than in Angus cows. Cortisol secretion after ACTH treatment was similar between Brahman and Angus cows. The cortisol response after ACTH challenge was positively correlated (r=0.68; P < 0.001) to the prechallenge area under the cortisol curve. Under optimal environmental conditions Brahman cows have a greater LH release and their anterior hypophysis is more sensitive to GnRH challenge than the Angus cows.     

Immunomodulatory effects of bovine colostrum in human peripheral blood mononuclear cells

Human and bovine colostrum (BC) contain a remarkable amount of bioactive substances, including antibodies towards many common pathogens of the intestinal and respiratory tract as well as growth factors, vitamins, cytokines and other proteic, lipidic and glucidic factors. In this study we investigated whether BC had any immunomodulatory effect on human peripheral blood mononuclear cells (PBMC) from healthy donors. To this aim we focused on the production of IL-12 and IFN-gamma, cytokines involved in the Th1 polarization required for a successful immune response towards intracellular pathogens, such as bacteria and viruses. BC induced a dose-dependent production of IL-12 by CD14+ monocytes, but was unable to induce IFN-gamma production. However, BC differentially affected stimuli-induced IFN-gamma production: it enhanced IFN-gamma in response to weak antigenic stimulation and it inhibited IFN-gamma in response to strong antigenic stimulation. These effects were not dose-dependent. We also measured PBMC proliferation, which was substantially unaffected by BC. Our data suggest that the Th1-promoting activity of BC could contribute, together with the antibodies, to the protective effect of BC on the offspring. BC could also represent an inexpensive therapeutic tool in prevention and treatment of several human microbial infections, including influenza.     

Effect of Supranutritional Organic Selenium Supplementation on Postpartum Blood Micronutrients,Antioxidants, Metabolites,and Inflammation Biomarkers in Selenium-Replete Dairy Cows

Dairy cows have increased nutritional requirements for antioxidants postpartum. Supranutritional organic Se supplementation may be beneficial because selenoproteins are involved in regulating oxidative stress and inflammation. Our objective was to determine whether feeding Se-yeast above requirements to Se-replete dairy cows during late gestation affects blood micronutrients, antioxidants, metabolites, and inflammation biomarkers postpartum. During the last 8-weeks before calving, dairy cows at a commercial farm were fed either 0 (control) or 105 mg Se-yeast once weekly (supranutritional Se-yeast), in addition to Na selenite at 0.3 mg Se/kg dry matter in their rations. Concentrations of whole-blood (WB) Se and serum Se, erythrocyte glutathione (GSH), and serum albumin, cholesterol, α-tocopherol, haptoglobin, serum amyloid A (SAA), calcium, magnesium, phosphorus, non-esterified fatty acids, and β-hydroxybutyrate were measured directly after calving, at 48 h, and 14 days of lactation in 10 cows of each group. Supranutritional Se-yeast supplementation affected indicators of antioxidant status and inflammation. Cows fed a supranutritional Se-yeast supplement during the last 8-weeks of gestation had higher Se concentrations in WB (overall 52 % higher) and serum (overall 36 % higher) at all-time points, had higher SAA concentrations at 48 h (98 % higher), had higher erythrocyte GSH (38 % higher) and serum albumin concentrations (6.6 % higher) at 14 days, and had lower serum cholesterol concentrations and higher α-tocopherol/cholesterol ratios at calving and at 48 h compared with control cows. In conclusion, feeding Se-replete cows during late gestation a supranutritional Se-yeast supplement improves antioxidant status and immune responses after calving without negatively impacting other micronutrients and energy status.     

Effects of infusion of GnRH pulses and level of body condition on ovarian function in postpartum beef cows

An experiment was conducted to test the hypothesis that postpartum anoestrus in beef cows is prolonged in cows in low body condition (BC) because they have a reduced LH pulse frequency compared with cows in high BC. Thirty-six multiparous Blue-Grey (White Shorthorn × Galloway) cows were fed so that they calved in either low (L) (BC score 2.07; SE 0.05; n = 24) or high (H) (BC score 2.81; SE 0.08; n = 12) body condition. They were then fed to maintain BC after calving. Twelve L cows were infused (i.v.) with 2 μg GnRH in 2 ml saline every 2 h from 5 to 7 weeks postpartum (LG) while the remaining L cows and all H cows were infused with saline only (LS and HS). Ovulations, as indicated by the presence of a morphologically normal corpus luteum, were recorded in one, one and ten of the cows of the LS, HS and LG groups, respectively. Mean LH concentrations and pulse frequencies were not affected by either GnRH treatment or BC but mean LH pulse amplitudes were lower (P < 0.05) in LG and LS cows than in HS cows at Week 5 and in LG cows at Week 6. At Week 7 postpartum, the numbers of small (3–7.9 mm diameter) and large (≥ 8 mm diameter) ovarian follicles, mean granulosa cell numbers per follicle and mean concentrations of LH receptors (pg per mg thecal and granulosa tissue) were not affected by GnRH treatment or BC. Granulosa cells from oestrogen active follicles of HS and LG cows secreted more oestradiol in vitro (P < 0.01) than cells from LS cows. However, there were no significant differences with treatment in the intrafollicular concentrations of oestradiol, testosterone or insulin-like growth factor-1. It was concluded that, since infusion of GnRH pulses enhanced both follicular steroidogenesis and the incidence of ovulation in low BC cows, the frequency of GnRH pulses is one determinant of follicle development in the postpartum cow. While H cows also exhibited a degree of enhancement of oestradiol synthesis by the granulosa cells of oestrogenic follicles, compared with L cows there was no difference in the LH pulse frequency or in the incidence of ovulation. It is concluded that there may be a threshold level of oestrogen synthesis by granulosa cells below which the final stages of follicle maturation and ovulation cannot be initiated, or that a high rate of oestradiol synthesis by this tissue is not the only factor mediating the effects of body condition on follicle development in the postpartum cow.     

The Hsp72 response in peri-parturient dairy cows: relationships with metabolic and immunological parameters

The study was aimed at assessing whether the peri-parturient period is associated with changes of intracellular and plasma inducible heat shock proteins (Hsp) 72 kDa molecular weight in dairy cows, and to establish possible relationships between Hsp72, metabolic, and immunological parameters subjected to changes around calving. The study was carried out on 35 healthy peri-parturient Holstein cows. Three, two, and one week before the expected calving, and 1, 2, 3, 4, and 5 weeks after calving, body conditions score (BCS) was measured and blood samples were collected to separate plasma and peripheral blood mononuclear cells (PBMC). Concentrations of Hsp72 in PBMC and plasma increased sharply after calving. In the post-calving period, BCS and plasma glucose declined, whereas plasma nonesterified fatty acids (NEFA) and tumor necrosis factor-alpha increased. The proliferative responses of PBMC to lipopolysaccharide (LPS) declined progressively after calving. The percentage of PBMC expressing CD14 receptors and Toll-like receptors (TLR)-4 increased and decreased in the early postpartum period, respectively. Correlation analysis revealed significant positive relationships between Hsp72 and NEFA, and between PBMC proliferation in response to LPS and the percentage of PBMC expressing TLR-4. Conversely, significant negative relationships were found between LPS-triggered proliferation of PBMC and both intracellular and plasma Hsp72. Literature data and changes of metabolic and immunological parameters reported herein authorize a few interpretative hypotheses and encourage further studies aimed at assessing possible cause and effect relationships between changes of PBMC and circulating Hsp72, metabolic, and immune parameters in dairy cows.     

Conception rate and reproductive function of dairy cows fed different fat sources

The objective of the experiment was to determine the effects of fat supplementation on cyclicity, progesterone concentration, follicular development, conception rate, embryo mortality, and plasma concentrations of prostalglandin F metabolite (PGFM) in cattle. The hypothesis of this experiment was that feeding flaxseed, which is a source rich in C18:3, would increase conception rate of dairy cows due to decreased plasma PGFM concentrations. A total of 138 lactating Holstein cows were allotted at calving to three groups of 46 cows, blocked for similar calving dates. Cows within each block were assigned to one of three isonitrogenous, isoenergetic, and isolipidic supplements based on either whole flaxseed (FLA), Megalac (MEG) or micronized soybeans (SOY). The diets were fed from calving to Day 50 of pregnancy for pregnant cows, or 120 day postpartum for those not diagnosed pregnant after AI. Detailed measurements of PGFM and follicle dynamics were only made on four cows for FLA and five cows for both MEG and SOY. The response in PGFM concentration following the oxytocin challenge administered around Week 11 of lactation was similar over time among treatments. Plasma progesterone concentrations from Days 17 to 21 of the estrous cycle starting around Week 9 of lactation and determined on a subsample of cows (n=for FLA and n=5 for both MEG and SOY) were higher for cows fed FLA than for those fed SOY (P=0.04) or MEG (P=0.06). Conception rates were similar among treatments. Total embryo mortality was lower (P=0.07) for cows fed FLA (0%) compared to those fed either MEG (15.4%) or SOY (8.0%). The mean size of the CL measured during a complete estrous cycle from Week 9 of lactation was smaller for cows fed SOY (16.3 mm) compared to those fed either FLA (19.1 mm) or MEG (18.3 mm). We inferred that pregnancy losses could be reduced by feeding whole flaxseed as a result of its effects on different factors such as modulation in concentration of progesterone and size of the CL.     

A comparison of natural service fertility of yearling and two-year-old bulls on pasture

The natural service fertility of yearling bulls on pasture was compared with that of 2-year-old bulls in a study (2 experiments) which involved a total of 748 composite breed cows over 3 breeding seasons. Other factors such as age of the cows, breed, year effects, and the sequence in which bulls of a particular age were assigned to cows were also evaluated. Pregnancy and calving rates and calving date were used as criteria for assessing fertility. The age of the bull, breed, year, or bull sequence did not significantly affect any of the traits studied. The age of cow was important in all the traits studied, with mature cows (5 years and older at breeding) calving earlier (P < 0.05) than the youngest group (2 years old at breeding) of cows, in both experiments. The pregnancy rate was also lower in the younger cows than in the mature cows in the first experiment. The results suggest that the age range of the cow herd is more important in determining herd fertility than the age of the bull or any of the parameters studied.     

Association between endometritis and endometrial cytokine expression in postpartum Holstein cows

The endometrium regulates the inflammatory response after infection by production and release of cytokines and chemokines. The objective was to compare gene expression of important pro-inflammatory (TNFα, IL-1β, IL-6) and anti-inflammatory (IL-10) cytokines, and the main neutrophil chemokine (IL-8), from calving to Week 7 after calving, in cows that developed endometritis and healthy control cows. Uterine biopsies were obtained at calving and at Weeks 1, 3, 5 and 7. Endometritis was evaluated at Week 5 by uterine lavage and cytology; cows with ≥ 10% neutrophils were considered to have endometritis. Real-time RT-PCR threshold values (Ct) were used to calculate the fold difference in gene expression, using the 2^−ddCt method, normalized to GAPDH and calibrated to the average dCt for all cows at calving. Serum IL-8 concentrations were measured with ELISA. The analysis included 28 cows (11 had endometritis) for the PCR data and 44 cows (20 had endometritis) for ELISA. Expression of the TNFα gene in uterine tissue was decreased in cows with endometritis compared to control cows at calving (P = 0.09) and at Week 1 (P = 0.05). Iterleukin-1β gene expression tended to be decreased (P = 0.08) in cows with endometritis compared to control cows at Week 1, but tended to be increased (P ≤ 0.10) at Weeks 5 and 7. Cows with endometritis had increased (P < 0.05) IL-6 gene expression at calving and at Week 7 compared to control cows. Interleukin-8 gene expression was increased (P = 0.03) in endometritic cows compared to control cows at Week 7. Uterine disease was not significantly associated with IL-10 gene expression. A lower local level of expression of pro-inflammatory cytokines in the endometrium soon after calving might impair activation of inflammation and clearance of bacteria, and lead to development of endometritis.