Effects of Feeding Dry Glycerol to Primiparous Holstein Dairy Cows on Follicular Development,Reproductive Performance and Metabolic Parameters Related to Fertility during the Early Post‐partum Period

This study examined the effects of dry glycerol supplementation on follicular growth, post‐partum interval to first ovulation, concentration of serum metabolites and hormones related to fertility, body condition score (BCS) and body weight (BW) in primiparous Holstein dairy cows. Sixty primiparous Holstein dairy cows were randomly assigned to two groups (control: n = 30 and glycerol supplemented: n = 30). Dry glycerol (250 g/day/cow) was fed as a top dressing to the common lactating total mixed ration (TMR) from parturition to 21 days post‐partum. Ovaries were examined four times using ultrasonography on days 13, 19, 25 and 36 post‐partum to determine ovarian follicular growth. Concentration of serum metabolites and hormones was determined weekly. Body condition score was evaluated weekly from weeks 1 to 5 after parturition, and BWs were recorded three times on days 1, 11 and 21 during the experimental period. The cows fed dry glycerol had more large follicles (p < 0.0001) and corpora lutea (CL) (p = 0.02) compared with the control cows. Days to the first ovulation (p = 0.06), days to first oestrus (p = 0.05), services per conception (p = 0.06) and days open (p = 0.004) were positively affected by dry glycerol supplementation. Serum concentration of glucose and insulin was higher in dry glycerol‐supplemented cows (p = 0.1; p = 0.06, respectively). Feeding glycerol had no effect on mean serum concentrations of β‐hydroxybutyrate, non‐esterified fatty acids and IGF‐1 during the experimental period. However, significant differences were observed at concentration of BHBA and IGF‐1 (p = 0.02 and p = 0.04, respectively) between two groups on day 21 after calving. The cows in the glycerol‐fed group had higher serum progesterone concentrations on days 33 (p = 0.007) and 36 (p = 0.004) after calving. Supplemented cows had lower body condition loss during weeks 1–5 after calving compared with the control cows (0.34 vs 0.41 BCS). In week 13 post‐partum, the proportion of cycling cows was 83.3 and 69.9% for those which received supplemented or non‐supplemented diet, respectively. These results demonstrated that feeding dry glycerol as a glucogenic supply may be useful to improve negative energy balance and reproductive efficiency in young cows which calve with high requirement of energy.     

Effects of pre‐partum milking of dairy cows on calcium metabolism at start of milking and at calving

This experiment studied the effect of pre‐partal milk removal on calcium metabolism at start of milking and at calving. Nine cows of the Swedish Red breed were milked for 1–7 days pre‐partum. The average milk yield at the first milking was 4.8 l, and the average yield the last day prior to calving was 13.4 l. Five cows were used as control cows and were only milked post‐partum. Samples of plasma and urine were taken to determine the effect of pre‐partum milking and calving on levels of calcium, magnesium, parathyroid hormone and plasma C‐terminal crosslinked telopeptide of type 1‐collagen (CTx), used as a marker of bone resorption. Pre‐partum milking resulted in a decrease in plasma calcium that was evident 2 days after the first milking. Parathyroid hormone increased at the same time, and CTx started to increase from 24 h after the first milking. There were no effects on plasma magnesium or urinary output of calcium or magnesium. The first week after calving, there were no differences between pre‐partum milked cows and control cows in plasma or urine variables, or in milk yield. In conclusion, pre‐partum milking activated the calcium‐restoring mechanisms but did not improve calcium status at calving.     

Effect of dietary potassium and anionic salts on acid–base and mineral status in periparturient cows

Dry cow diets based on grassland forage from intensive production contain high amounts of K and could be responsible for a reduced ability to maintain Ca homoeostasis. The aim of this study was to determine whether a moderate anionic salt supplementation to a forage‐based pre‐calving diet with varying native K content affects the mineral and acid–base status in transition cows. Twenty‐four dry and pregnant Holstein cows, without antecedent episodes of clinical hypocalcemia, were assigned to two diets during the last 4 weeks before estimated calving date. Twelve cows were fed a hay‐based diet low in K (18 g K/kg DM), and 12, a hay‐based diet high in K (35 g K/kg DM). Within each diet, six cows received anionic salts during the last 2 weeks before the estimated calving day. After calving, all cows received the high K diet ad libitum. Blood samples were taken daily from day 11 pre‐partum to day 5 post‐partum. Urine samples were taken on days 7 and 2 pre‐partum and on day 2 post‐partum. The anionic salt did not alter feed intake during the pre‐partum period. Serum Ca was not influenced by the dietary treatments. Feeding pre‐partum diets with low K concentrations induced a reduced metabolic alkalotic charge, as indicated by reduced pre‐partum urinary base–acid quotient. Transition cows fed the low K diet including anionic salts induced a mild metabolic acidosis before calving, as indicated by higher urinary Ca, lower urinary pH and net acid–base excretion. Although serum Ca during the post‐partum period was not affected by dietary treatment, feeding a low K diet moderately supplemented with anionic salts to reach a dietary cation–anion difference close to zero permitted to obtain a metabolic response in periparturient cows without altering the dry matter intake.     

Physiological changes in the adenosine deaminase activity,antioxidant and inflammatory parameters in pregnant cows and at post‐partum

The aim of this study was to evaluate the behaviour of adenosine deaminase (ADA) activity, as well as its participation in the immunomodulation of pregnant cows. Thus, sixteen cows were divided into two groups (A and B): the group A was composed by cows not pregnant (n = 8), while the group B was composed by pregnant cows (n = 8). Serum levels of interleukin‐10 (IL‐10), IL‐6, tumour necrosis factor alpha (TNF‐α), reactive oxygen species (ROS) and C‐reactive protein (CRP), as well as ADA and glutathione S‐transferase (GST) activities, were measured on five sampling times (3, 5, 7 and 8 months of gestation, and soon after calving). Serum ADA activity was similar throughout the experiment in the cows belonging to the group A, but its activity increased during the experiment in cows from the group B, that is it was lower in the third and fifth months of pregnancy, and higher on months 7, 8 and after calving when compared to the group A. TNF‐α and IL‐6 serum levels were lower in pregnant cows compared to non‐pregnant animals; however, they significantly increased after calving. Serum levels of IL‐10 increased after 8 months of gestation, but it reduced after calving when compared to the group A, while CRP increased on month 8 of gestation and after calving compared to the group A. Pregnant cows showed lower serum ROS levels on months 3, 5 and 7 of gestation, and higher levels at the post‐partum. Serum GST activity was higher on month 5 of gestation in pregnant cows, but it was lower on months 7, 8 and in the post‐partum compared to the group A. Based on these evidence, we concluded that ADA activity and the others mediators or inflammatory modulators have important role in the maintenance of cow′s gestation due to their immunomodulatory effects.     

Gonadotropin‐Releasing Hormone Administration to Dairy Cows without a Corpus Luteum 4 Weeks after Calving Increases Reproductive Performance

This field study investigated whether the administration of a single dose of gonadotropin‐releasing hormone (GnRH) to dairy cows without a corpus luteum (CL) 4 weeks after calving can improve reproductive performance. Holstein dairy cows underwent ultrasonography to assess the presence of ovarian structures at 29.2 ± 5.2 days post‐partum, and cows were divided into two main groups based on the presence (CL group, n = 230) or absence (non‐CL group, n = 460) of a CL. The non‐CL group was further randomly divided into two subgroups based on the administration of GnRH (non‐CL GnRH group, n = 230) or no GnRH (non‐CL control group, n = 230). Subsets of cows from non‐CL control (n = 166) and non‐CL GnRH (n = 175) groups received a second ultrasonography at 44.5 ± 5.4 days post‐partum to assess CL formation. The percentage of cows with CL at the second ultrasonography was greater in the non‐CL GnRH group (70.9%) than in the non‐CL control group (53.0%, p = 0.0006). The hazard of the first post‐partum insemination by 150 days in milk (DIM) was higher in the CL group than in the non‐CL control group (hazard ratio [HR]: 1.36, p = 0.001). The probability of a pregnancy to the first insemination was higher in non‐CL GnRH (odds ratio [OR]: 1.50, p = 0.04) and CL groups (OR: 1.55, p = 0.03) compared to the non‐CL control group. Furthermore, the hazard of pregnancy by 210 DIM was higher in non‐CL GnRH (HR: 1.30, p = 0.01) and CL (HR: 1.51, p = 0.0001) groups than in the non‐CL control group. In conclusion, administration of GnRH to dairy cows without a CL 4 weeks after calving was associated with an increase in ovulation and improved reproductive performance.     

Metabolic and luteal function in winter‐calving Spanish beef cows as affected by calf management and breed

This experiment aimed at evaluating the effect of calf management and breed on the metabolic and luteal function of post‐partum beef cows fed at maintenance. Fifty multiparous cows, 22 Parda de Montaña (PA) and 28 Pirenaica (PI), were assigned to either suckling once‐daily for 30 min (RESTR) or ad libitum (ADLIB) from the day after calving. Blood samples were collected to analyse metabolites [non‐esterified fatty acids (NEFA), β‐hydroxybutyrate, total protein and urea)], insulin‐like growth factor‐I (IGF‐I) and progesterone (P4) at different intervals. Cows from RESTR maintained their live‐weight (LW) over the first 3 months post‐partum, whereas ADLIB cows lost nearly 4% LW. Both genotypes showed similar LW gains during this period (p > 0.10). Calf daily gains were lower in RESTR than in ADLIB treatment (p < 0.05), but similar across breeds (p > 0.10). Milk and lactose production were lower in RESTR cows than in ADLIB (p < 0.05). Milk and protein yield were greater in PA than in PI breed (p < 0.05). Serum NEFA, total protein and urea were higher in PI cows suckling ADLIB than in the rest (p < 0.05). Cows from PI breed had greater NEFA values than PA ones on the first week post‐partum (p < 0.001). Circulating IGF‐I was not affected by suckling frequency, breed nor their interaction (p > 0.10). Suckling frequency, but not breed, affected the interval from calving to first ovulation (p < 0.001), being shorter in RESTR than in ADLIB cows. In conclusion, the ad libitum suckling practice improved cow milk yield and offspring gain compared to once‐daily suckling for 30 min from the day after calving, at the expense of impairing the onset of cyclicity. The effect of calf management was confounded with breed on the studied blood biochemical constituents, but any of these metabolites influenced the role of endocrine IGF‐I in these genotypes.     

The Effect of Lactation on Post‐Partum Uterine Involution in Holstein Dairy Cows

The objective was to examine the effect of lactation on uterine involution in post‐partum dairy cows. Holstein primiparous cows were used (n = 19, mean age: 3.9 ± 0.1 years). At calving, cows were randomly assigned to one of two treatment groups, lactating (n = 11) or non‐lactating (i.e. dried off at calving, n = 8). Examination of the reproductive tract was carried out by ultrasonography twice weekly until week 7 post‐partum. Blood samples were collected twice weekly for the analysis of progesterone to indicate the resumption of cyclicity and metabolites indicative of energy status. Uterine involution was assessed in terms of size of the uterine horns, uterine body diameter and uterine fluid volume as assessed by the amount of non‐echogenic material measured by ultrasound and position of the uterus. Vaginal mucous score was taken on day 28 post‐partum for the assessment of uterine inflammation. Resumption of cyclicity (serum progesterone > 1 ng/ml) had occurred in both groups on average by day 21 post‐partum. Concentrations of non‐esterified fatty acids and beta‐hydroxybutyrate were higher, whereas concentrations of glucose, insulin and IGF‐1 were lower (p < 0.05) in lactating compared to non‐lactating cows. Lactating cows had a smaller mean uterine body diameter (p < 0.05) than non‐lactating cows from days 28 to 42 post‐partum (day 28: 20.2 ± 1.3 vs 24.9 ± 1.5 mm, respectively) and had a lower mean uterine fluid volume up to day 49 (p < 0.05). By day 49, there was no difference in uterine diameter (15.2 ± 1.8 vs 15.2 ± 1.6 mm) or uterine fluid volume (0.11 ± 0.38 vs 0.18 ± 0.46) between lactating and non‐lactating cows, respectively. Vaginal mucous score revealed no evidence of uterine inflammation in either group. In conclusion, while lactation induced significant alterations in metabolic status, it did not have a major effect on the rate of uterine involution as defined in this study.     

Endocannabinoids as endometrial inflammatory markers in lactating Holstein cows

The objective of this study was to consider endocannabinoid system as inflammatory markers in bovine endometrium to better understand the role of this system in regulating many of the functions that are related to inflammatory condition. At day 26 post‐partum, fourteen cows were divided into two groups depending on the inflammatory condition: 1‐ subclinical endometritis (n = 7, with purulent or mucopurulent uterine discharge detectable in the vagina) and 2‐ healthy (n = 7, No (muco)) purulent discharge. Blood samples were collected at 26 and 30 days relative to calving to determine plasma tumour necrosis factor (TNF) and lipopolysaccharide‐binding protein (LBP) concentrations; moreover, uterine biopsy was carried out on day 26 post‐partum to measure mRNA abundance of TNF, interleukin‐1B (IL1B), interleukin‐6 (IL‐6), C‐X‐C motif chemokine ligand 8 (CXCL8), endocannabinoid receptor (CNR2), N‐acyl phosphatidylethanolamine phospholipase D (NAPEPLD), fatty acid amide hydrolase (FAAH), N‐acylethanolamine acid amidase (NAAA) and monoglyceride lipase (MGLL) by real‐time PCR. Results showed mean plasma concentrations of TNF and LBP were lower in healthy cows compared to subclinical endometritis cows (p < .05). Relative mRNA expression for NAAA and FAAH was decreased (p < .05), and relative mRNA expression for CNR2 and NAPEPLD increased in cows with subclinical endometritis compared to healthy cows. In conclusion, relative mRNA expression of TNF, IL1B and CXCL8 and plasma concentration of LBP increased during inflammatory condition along with decreased endocannabinoids hydrolyzing enzyme (NAAA and FAAH), increased enzymes that synthesize endocannabinoids (NAPEPLD) and relative gene expression of the endocannabinoid receptor; together, these contribute to increased endocannabinoids levels during inflammation. Overall, we provide evidence that endocannabinoid system is altered in endometrium tissue during inflammation through increased mRNA expression of CNR2 and synthesis enzyme and decreased mRNA expression of hydrolyzing enzymes interfere with pro‐cytokine production and signalling, which may interfere with the onset and progression of inflammation.     

Expression profiles of immunity and reproductive genes during transition period in Holstein cattle

The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real‐time PCR to determine mRNA levels for the immune‐related genes TLR 2, 4, 6, 7 and β‐defensin 5 in addition to the reproduction‐related genes prolactin and IGF‐I. Results showed significant up‐regulation of pro‐inflammatory‐selected genes, TLR 4, 6 7 and β‐defensin 5 at the third‐week post‐calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up‐regulated at calving and 1 week after parturition and then down‐regulated again at second and third week. Prolactin and IGF‐I genes expression levels were significantly and gradually increased mainly post‐partum. This research highlights that the expression patterns of TLRs, BNBD5, PRL and IGF‐I could be biomarkers to follow up immune and reproductive status of dairy cow at peri‐parturient period to predict the most susceptible risk time for disease incidence and to build up management protocol.     

Short‐term methionine supplementation during the early post‐partum period in primiparous rabbits improves prolificacy associated with an increase in serum concentrations of IGF‐I

The aim of this study was to evaluate the effects of methionine supplementation on energy metabolism and reproductive performance during the early post‐partum period in primiparous does. Forty nulliparous New Zealand White does were used. Females were randomized in two groups at calving: the control group (n = 20) was fed with the basal diet, and the methionine group (n = 20) was fed the basal diet plus 1 g/animal/day of methionine from the day of calving to 4 days post‐partum. Results showed that methionine supplementation increased (p = 0.032) the concentration of insulin‐like growth factor‐1 with respect to control group 4 days post‐partum. It similarly increased the prolificacy (p = 0.03), the number of kits born alive per litter (p = 0.06) and the body gain weight of the litter during supplementation (p = 0.035). These results were observed despite the does in the methionine group having a deeper negative energy balance than the does in the control group. Finally, methionine supplementation did not affect receptivity (p = 0.23), fertility (p = 0.49), the number of kits born dead per litter (p = 0.86) insulin and metabolites as glucose, non‐esterified fatty acids and triglycerides. In conclusion, our results show that methionine supplementation during the first 4 days of the post‐partum period in rabbits increases total litter size and the corporal weight of kits and is associated with an increase in blood concentration of IGF‐1.     

Effect of pre‐ and post‐partum supplementation with lipid‐encapsulated conjugated linoleic acid on milk yield and metabolic status in multiparous high‐producing dairy cows

We evaluated the lactation performance, liver lipid content and plasma metabolites indicating the energy balance of dairy cows supplemented with conjugated linoleic acid (CLA) pre‐ and post‐partum (PP) vs. only PP. A total of 60 cows were divided into three groups (n = 20). Daily diet of cows was supplemented with 14 g of CLA (7 g cis‐9, trans‐11 and 7 g trans‐10, cis‐12 isomers) from week 3 before the expected date of calving (group CLA1), or from the day of calving (group CLA2) until 77–91 days PP. Control cows were fed an isocaloric, isonitrogenous and isolipidic diet without CLA. Between week 3 and week 6 PP, the milk yield of cows in both CLA‐treated groups was approximately 4.5 kg higher (p < 0.05) than in control. Milk fat concentrations decreased from week 3 and were lower in both CLA groups than in control (p < 0.01). Body condition score loss was lower (p < 0.05) in the CLA1 than in the control group on week 5 PP. By week 11 PP, the body condition of both CLA1 and CLA2 groups exceeded that of control. Plasma non‐esterified fatty acid was lower in CLA1 compared to CLA2 and control during the early PP period (p < 0.05), while this difference faded away by the late PP period. Beta‐hydroxybutyrate (BHBA) increased rapidly in all groups following calving. In CLA1 group, it began to decrease sooner than in CLA2 and control. The prevalence of subclinical ketosis (BHBA > 1.2 mm ) was lower in CLA1 group than in CLA2 and control (p < 0.05). Liver biopsy analyses showed that CLA1 treatment decreased (p < 0.05) the total lipid content of liver compared to control at week 5 after calving. Our results show that CLA supplementation is more efficient in alleviating body mass mobilization and decreasing the incidence of subclinical ketosis when applied as early as 3 weeks before calving than started feeding after calving.     

Metabolic parameters and their relationship to energy balance in multiparous Simmental,Brown Swiss and Holstein cows in the periparturient period as influenced by energy supply pre‐ and post‐calving

A study was conducted to evaluate the effects of three energy supply (E) levels [low (L), medium (M), high (H)], both pre‐partum (PRE) and post‐partum (POST), and their interactions on metabolic parameters and energy balance (EB) in dairy cows of three breeds. In both phases, E levels applied to a total of 81 multiparous cows of breeds Simmental (SI), Brown Swiss (BS) and Holstein–Friesian (HF; n = 27 for each breed) were 75%, 100% and 125% of recommendations of the German Society of Nutrition Physiology, using a 3 × 3 factorial arrangement of treatments. During the pre‐calving period, serum concentrations of non‐esterified fatty acids (NEFA) were higher for L_PRE cows, and glucose concentrations were elevated for H_PRE cows. During the lactation period, NEFA concentrations were greatest for treatment L_POST. Mean concentrations of β‐hydroxybutyrate (BHB) were highest for cows of the L_POST treatment, intermediate for M_POST and lowest for H_POST. Glucose concentrations were lower for L_POST cows. SI cows had lower BHB concentrations both pre‐ and post‐calving and higher glucose concentrations during early lactation than the other breeds. BHB concentration POST was highest for BS cows. Restricted feeding PRE resulted in a better energy status of cows fed above energy requirements POST (E_PRE × E_POST interaction). HF cows had a higher EB pre‐calving, whereas SI cows had a less negative EB during early lactation, compared with the other breeds respectively. Correlations of serum NEFA, BHB and glucose concentrations with EB were strongest during the transition period. Results suggest that controlling energy intake during the dry period might be advantageous for the energy status of dairy cows after calving, whereas energy restriction in early lactation leads to metabolic stress. Evidence is provided of a clear relationship between EB and the blood metabolites NEFA and BHB, especially in the transition period.     

Husbandry Factors and the Resumption of Luteal Activity in Open and Zero‐Grazed Dairy Cows in Urban and Peri‐Urban Kampala,Uganda

The study investigated the influence of selected husbandry factors on interval to resumption of post‐partum cyclicity among dairy cows in urban and peri‐urban Kampala. A prospective study of 85 day post‐partum period of 59 dairy cows in open (n = 38) and zero grazing (n = 21) systems was conducted on 24 farms. Cows of parity 1–6 were recruited starting 15–30 days post‐partum. Progesterone (P4) content in milk taken at 10–12 day intervals was analysed using ELISA. The cow P4 profiles were classified into ‘normal’ (< 56 days), ‘delayed’ (> 56 days), ‘ceased’ or ‘prolonged’ (if started < 56 days but with abnormal P4 displays) resumption of luteal activity and tested for association with husbandry and cow factors. Of the 59 cows, luteal activity in 81.4% resumed normally and in 18.6%, delayed. Only 23.7% maintained regular luteal activity, while the others had ceased (10.2%), prolonged (37.3%) or unclear luteal activity (20.3%). There were no differences between open and zero‐grazed cows. Milk production was higher (p < 0.05) in zero than open grazing, in urban than peri‐urban and in cows fed on brew waste (p < 0.001) compared with mill products and banana peels. Results suggest that luteal activity resumes normally in a majority of cows, although only a minority experienced continued normal cyclicity once ovulation had occurred, in the two farming systems irrespective of feed supplements or water, and that supplementing with brew waste is beneficial for milk production.     

Effect of supplementation of vitamin E,copper and zinc on the in vitro phagocytic activity and lymphocyte proliferation index of peripartum Sahiwal (Bos indicus) cows

To study the effect of vitamin E (VE), copper (Cu) and zinc (Zn) supplementation on the in vitro phagocytic activity (PA) and lymphocyte proliferation response (LPR) of blood neutrophils and lymphocytes, thirty Sahiwal pregnant cows (six in each group) in their late gestation at 30 days before the expected date of calving were selected from the NDRI experimental herd and supplemented with various micronutrients from 30 days before calving to 45 days after calving. Cows were supplemented individually with VE (1000 IU/cow/day), Cu (20 ppm/cow/day) and Zn (80 ppm/cow/day) and also with a combination of VE, Cu and Zn to study cumulative effect of all micronutrients. One group without any supplementation acted as a control. Blood neutrophils and lymphocytes were isolated and studied for their PA and LPR. Supplementation of micronutrients like VE, Cu, Zn and a combination of all these nutrients significantly (p < 0.01) increased the PA of experimental cows as compared to control (unsupplemented) cows during the pre‐partum period. During post‐partum, all the micronutrients (VE, Cu, Zn and their combination) showed a significant (p < 0.01) increase in the PA of experimental cows as compared to control cows. Of all the groups, significant (p < 0.01) and maximum PA was observed in the combination group followed by Zn‐supplemented group during both the pre‐ and post‐partum period. A significant (p < 0.01) increase in LPR of B lymphocytes was observed in combination‐supplemented group during the pre‐partum period and during both the pre‐ and post‐partum period in the Cu‐supplemented group.     

Dominant follicles development and estradiol‐17β concentrations in non‐ovulating and ovulating post‐partum Bulgarian Murrah buffaloes

This study was designed to evaluate the dominant follicles development and the estradiol‐17β concentrations in non‐ovulating and ovulating post‐partum buffaloes. Sixteen Bulgarian Murrah buffaloes were submitted to transrectal ultrasonographic examination from the 1st post‐partum day until day 50, 3 days apart. The follicular diameter of the different categories of follicles and the ovulations was recorded. The animals were allocated into two groups: I (n = 6) non‐ovulating and II (n = 10) ovulating buffaloes. Serum estradiol‐17β concentrations on the days for dominant follicle registration were measured by enzyme‐linked immunosorbent assay. The results were statistically processed by analysis of variance, non‐parametric and correlation analysis. The mean intervals between calving and first dominant follicle detection differed significantly (p < .05) among the groups (19.5 ± 6.2 vs. 13.8 ± 5.1 days), while the mean intervals between registered dominant follicles from two successive waves were comparable. The mean follicular diameters for the same category follicles in both groups were similar. Different estradiol‐17β concentrations (p < .05) for the first dominant follicle between non‐ovulating (23.5 ± 7.0 pg/ml) and ovulating (33.3 ± 8.4 pg/ml) buffaloes were determined. The cumulative percentages of buffaloes with firstly detected dominant follicle and ovulating animals correlated positively (r ≥ .84; p < .05) to post‐partum days. In conclusion, non‐ovulating and ovulating post‐partum Bulgarian Murrah buffaloes showed differences in the development of the first dominant follicle and estradiol‐17β concentrations during the time of dominant follicles detection.     

Polymorphism in 5′ untranslated region of heat‐shock protein 70 gene as marker of post‐partum anoestrus in Murrah buffaloes

The enormous production potential of buffaloes has never been accomplished due to various reproductive insufficiencies. Among them, post‐partum anoestrus, a multifactorial disorder, is predominant but any genetic association is yet to be established. This study focused to identify novel polymorphisms in heat‐shock protein 70 (HSP70) gene and its possible association with post‐partum anoestrus in Murrah buffaloes. A 579‐bp fragment from 5′ untranslated region of HSP70 gene was amplified by polymerase chain reaction from blood genomic DNA of 614 animals maintained under similar management conditions. In phase‐I experiment, custom sequencing and restriction enzyme (RE) digestion of the amplified fragment were performed in 40 buffaloes with similar post‐partum oestrous conditions over previous consecutive three or more gestations—20 animals each showing post‐partum anoestrus (>120 days after parturition) and normal cyclicity (<65 days after parturition). While in phase‐II experiment, herd screening by RE analysis was performed in remaining 574 animals. Four transversions at T‐75G, C+31G, T+38G and C+97A and three transition mutations at T‐153C, T+33C and A+44G positions were observed. Polymorphism at T+38G site revealed significant (p < .05) variation, where homozygous G was present only in post‐partum anoestrous animals while nucleotide T was present randomly in both groups of phase‐I animals whereas phase‐II experiments revealed homozygous G in 55 animals. Regression analysis in relation to average post‐partum interval against genotypic frequencies at T+38G also depicted significant association. HSP70 gene polymorphism at T+38G position can therefore be used as genetic marker for excluding probable post‐partum anoestrous buffaloes from herd for breeding programmes.     

OC10A Negative Energy Balance but not Suckling Frequency (ad libitum,Twice or Once Daily) Modules Ovarian Resumption in Parda de Montaña Cattle Breed

In extensive systems, beef cows should be continuously with their calves to optimise pasture use but this practice can lengthen post‐partum interval (PPI). A study was conducted to determine the influence of suckling frequency on Parda de Montaña cow‐calf performance when cows are fed 70% energy requirements during lactation (outdoor winter conditions simulation). Thirty‐six autumn‐calving cows with similar body condition at calving (2.57) were assigned to three suckling systems [ad libitum (AL), twice (2D) or once daily (1D) for 30 min]. Blood samples were collected twice a week to analyse progesterone concentrations by RIA. Cows lost similar weight until start of mating period (day 52 post‐partum; ?0.583, –0.513, –0.520 kg/day in AL, 2D and 1D). Standard milk yield was higher in AL than the rest (8.9, 6.2, 7.0 kg; p < 0.05), which was reflected on greater calf gain (0.895, 0.752, 0.676 kg/day; p < 0.05) and larger cow weight loss within 90 days post‐partum in this treatment (–0.345, –0.188, –0.083 kg/day; p < 0.05). Suckling system did not affect either PPI (69.6, 89.1, 65.5 days) or cows cycling within 90 days post‐partum (55, 46, 58%), which may compromise the target calving interval. In Parda de Montaña breed fed moderately pre‐calving and undernourished during lactation, restricted suckling did not favour ovarian activity resumption, but post‐partum subnutrition delayed about 40 days PPI observed in similar body condition at calving and calf management (Sanz et al., 2003; Anim Reprod Sci 79: 57–69)     

Evaporative cooling in late gestation heat‐stressed Murrah buffaloes increases efficiency of next reproductive cycle

Evaporative cooling during late gestation period improves post‐partum reproductive performance in Murrah buffaloes. To prove this hypothesis, sixteen pregnant dry Murrah buffaloes at sixty days pre‐partum were selected and divided into two groups of eight animals each. Group 1 of buffaloes (Cooled/CL) was managed under fan and mist cooling during dry period, whereas second group of buffaloes (non‐cooled/NCL) remained without the provision of cooling. After parturition, all the animals were managed under evaporative cooling till the end of experimental period. Reproductive performance in cooled (CL) and non‐cooled (NCL) groups, respectively, viz. 1st and 2nd ovulation from calving (48.63 ± 2.41, 69.25 ± 2.34 days and 57.75 ± 3.35, 93.63 ± 2.84 days); calving to conception interval (117.88 ± 4.21 days and 117.88± 4.21 days); conception rate (87.5% ± 2.16% and 57% ± 2.26%); and follicular diameter at the time of 1st and 2nd ovulation (14.84 ± 0.16, 15.75 ± 0.13 mm and 12.65 ± 0.13, 13.35 ± 0.11 mm) varied significantly (p < .05). Total peak oestrogen concentration was significantly (p < .05) higher in cooled (26.7 ± 1.32 pg/ml) relative to non‐cooled (20.7 ± 1.22 pg/ml) buffaloes. Time from onset of oestrus to ovulation varied significantly (p < .05) in cooled (32 ± 2.22 hr) and non‐cooled (40 ± 2.86 hr) buffaloes. The peak progesterone concentration reached to (4.25 ng/ml) in cooled group and (4.16 ng/ml) in non‐cooled group after first ovulation.     

Luteinizing hormone and growth hormone secretion in early lactating Spanish beef cows

The episodic release of luteinizing hormone (LH) and growth hormones (GH) was studied in three suckling regimens and two breeds of Spanish suckled cows. Parda de Montaña (PA) cows (n = 21) were assigned to once‐daily, twice‐daily or ad libitum (ADLIB) suckling. Pirenaica (PI) cows (n = 7) were used to evaluate the breed effect in twice‐daily suckling. Coccygeal blood samples were collected twice weekly during lactation to determine the interval from calving to first ovulation through peripheral progesterone. On day 32 ± 3 post‐partum, jugular blood samples were drawn at 15 min intervals during 8 h to analyse circulating LH and GH. The interval to first ovulation was greater in PA cows suckling ADLIB than in restricted suckling treatment (RESTR1), whereas in RESTR2 it did not differ from the other two treatments. There were no differences between PA and PI cows in the interval to first ovulation. RESTR1 cows showed a tendency to have shorter LH peak widths than ADLIB cows. PA cows showed a tendency to have longer LH peak widths than their PI counterparts. There were no differences across treatments or breeds in any of the GH measures of secretion. The LH release was more affected by breed than by suckling frequency, whereas that of GH was not influenced by any of these parameters. The variables that best allowed discrimination between ADLIB and restricted nursing systems were the interval to post‐partum first ovulation, LH peak number and the mean GH concentration.     

Peripartal calcium homoeostasis of multiparous dairy cows fed rumen‐protected rice bran or a lowered dietary cation/anion balance diet before calving

Milk fever is one of the most important metabolic diseases in dairy cattle. Reducing the dietary cation/anion balance (DCAD) with anionic salts is a common prevention strategy. However, many small European farms cannot use total mixed rations (TMR) in the close‐up period. Including anionic salts in compound feeds can result in feed refusals and moderate inclusions to preserve feed palatability results in insufficient DCAD reduction. Rumen‐protected rice bran induces the adaptation of Ca metabolism in dairy cows by a reduction of Ca intake and by a reduction of the availability of dietary Ca. In the presence of a negative control, rumen‐protected rice bran (2.8 kg/day) was compared with a lowered DCAD diet (from 269 to 4 meq/kg DM) in their effect to prevent milk fever. In a randomized block design, 45 multiparous Holstein cows joined the trial sequentially from 21 days before the expected calving date and were observed until the 8th week of lactation. Feed and nutrient intakes were recorded, and Ca, P, Mg in serum and urine, urine pH, serum NEFA and milk production in early lactation were compared. Feeding rumen‐protected rice bran before calving improved the recovery of calcaemia after calving and had a positive effect on DMI after calving. The moderately low DCAD diet did not positively influence serum Ca at calving. Calcaemia recovered even later than in control, and cows showed reduced DMI post‐calving and higher NEFA levels in the first 36 h after calving. This moderate reduction of DCAD did not provide an intermediate prevention level indicating that DCAD needs to be reduced to the recommended levels to prevent milk fever. Rumen‐protected rice bran may be a suitable feed to reduce hypocalcaemia post‐partum and can be included in pre‐calving compound feeds representing a palatable alternative to anionic salts.