T cells play a central role in specific immunity; their populations and phenotypes could be affected by number of lactation in high-yielding dairy cows. To investigate the effects of parity on the dynamics of T lymphocytes, lymphoproliferative capacity, T lymphocyte subsets and CD4+/CD8+ ratio were studied in peripheral blood of primiparous and pluriparous dairy cows during mid–late lactation. A non-radioactive technique was also adapted for a detailed lymphoproliferation assay. Compared with the primiparous cows, the pluriparous cows exhibited weaker lymphoproliferative activity, larger number of CD4+ cells and substantially greater CD4+/CD8+ ratio in their blood circulation. The increase of the CD4+/CD8+ ratio in the blood of pluriparous dairy cows was mainly due to the rise in the proportion of CD4+ cells and decline in the proportion of CD8+ cells. This increase of the CD4+/CD8+ ratio coincided with the decrease of mitogen-induced proliferation capacity of T lymphocytes. Of four lymphocyte divisions or generations during the lymphoproliferation assay, maximal lymphocyte proliferation capacity at generation 3 in primiparous cows was markedly greater than in pluriparous cows. With an alternatively safer, faster and more reproducible assay (compared with 3H-thymidine scintillation assay) we showed for the first time that aging in dairy cows leads to a decreased mitogen-induced lymphoproliferation and disturbed proportion between CD4+ and CD8+ T cells. This CD4+-CD8+ imbalance together with diminished lymphoproliferative capacity may lead to a weaker T cytotoxic-mediated immunity and increased susceptibility to infectious diseases in pluriparous lactating cows. Our study also emphasizes further application of the methods in farm animals.

We examined the heat stability, somatic cell count (SCC), pH, fat, protein and lactose content of milk from goats during the oestrous period, in order to investigate evidence of possible oestrus effects on milk physical and chemical properties. Goats free from mammary infections were ranked on average SCC from three tests so that they could be stratified randomly in pairs to synchronized oestrus or left as unsynchronized non-oestrus controls. The synchronisation consisted of insertion of an intravaginal progesterone-releasing device for 17 d, and introduction of the bucks the day of the device removal (D0). The repeated measurements analysis of variance model included the fixed effects of the experimental group (oestrus or control) and day and the corresponding interaction and also the random effect of doe. Reduced milk-heat stability, increased SCC, increased protein content and reduced pH were found in the milk samples of the oestrus group on D1, 2 and 3. The fat and lactose content of the milk was not affected by oestrus. These data indicate that the milk of goats during the mating period has reduced heat stability and, therefore, that dilution into bulk tanks should be recommended to avoid clotting when milk is intended for high thermal treatment.

The level of gelatinases in surrounding body fluids of actively remodelling tissue is indicative of basement membrane and extracellular matrix degradation under various physiological and pathological circumstances. To elucidate the association of gelatinase with mammary tissue remodelling during gradual or acute involution, in the first trial, goats milked twice daily (lactation) and goats receiving decreased milking frequency (involution) served to provide a total of 12 milk samples and 11 mammary secretion samples, respectively. In the second trial, 6 cows served to provide samples of dry secretion in 3 consecutive weeks immediately following milk stasis. Gelatin zymography was applied for gelatinase phenotyping and quantification on milk, plasma and the degranulation medium/lysate of milk somatic cells. Results indicated that the most prevalent gelatinase subtype switched from gelatinase A in milk to gelatinase B in involution secretion. Mammary secretion of goats during involution contained marginally higher protein level, significantly lower casein ratio and greater specific capacity of gelatinase B compared with those of milk during lactation. Specific capacities of gelatinases A and B in plasma of goats were similar during lactation and involution, while gelatinase B capacity in degranulation medium/lysates based on unit number of goat somatic cell was significantly higher during involution than during lactation. Milk stasis of cows induced a significant increase in specific capacity of gelatinase B, but not gelatinase A, of dry secretion up to the third week. Results of both trials agree that regional selective accretion of gelatinase B in milk might have played a role in mammary tissue remodelling during involution induced by either decreasing milking frequency or milk stasis. It is suggested that infiltrated polymorphonuclear neutrophils are one of the potential contributors responsible for the accumulation of gelatinase B during involution.

Mastitis is the most frequent and prevalent production disease in dairy herds in developed countries. Based on a milk somatic cell count (SCC) of either >300 000 or <200 000 cells/ml in this study, we defined the quarter as either inflamed or uninflamed, respectively. The electrical conductivity (EC) of milk was used as an indicator of udder epithelial cell damage. We determined the amount of H2O2 produced by utilizing a small molecular weight compound in milk, and examined the characteristics of H2O2 production and EC in milk from inflamed and uninflamed quarters. In cows with milk of delivery grade (control population), H2O2 production and EC were 3·6±1·3 nmol/ml and 5·4±0·4 mS/cm (mean±sd), respectively. In 37 inflamed quarter milk samples, the production of H2O2 was 1·9±1·0 nmol/ml and was significantly smaller than that in the control population (P<0·01). Production of H2O2 was moderately but significantly correlated with EC (r<−0·71). In 20 cows with inflamed quarters, the production of H2O2 in milk from inflamed quarters was significantly smaller than that in milk from uninflamed quarters (P<0·01). In 18 out of 20 cows, milk from inflamed quarters showed the smallest H2O2 production among all tested quarters in each cow. We conclude that inflammation caused a decrease in H2O2 production in milk. In this study, we present parameters for evaluating the lactoperoxidase/H2O2/thiocyanate antibacterial defence system in bovine milk.

The effects of ultra-high pressure homogenization (UHPH) on skim milk yogurt making properties were investigated. UHPH-treated milk was compared with conventionally homogenised (15 MPa) heat-treated skim milk (90°C for 90 s), and to skim milk treated under the same thermal conditions but fortified with 3% skim milk powder. Results of the present study showed that UHPH is capable of reducing skim milk particle size which leads to the formation of finer dispersions than those obtained by conventional homogenisation combined with heat treatment. In addition, results involving coagulation properties and yogurt characteristics reflected that, when increasing UHPH pressure conditions some parameters such as density of the gel, aggregation rate and water retention are improved.

A method for the estimation of urea in milk using ammonia electrode is described. Urea is first degraded by urease enzyme into ammonium ion and carbon dioxide at neutral pH. The ammonium ion is then converted into ammonia at alkaline pH. A linear inverse relationship was observed between logarithmic concentration of ammonia or urea and electrode response. Repeatability, expressed as a coefficient of variation, was 1·77% at a level of 8·92 mm-urea in milk. The method was validated in milk samples spiked with between 2×10−3 and 10×10−3 m-urea and recovery of added urea was quantitative. Whereas, preservative sodium azide at 0·5 g/l or 2 g/l level did not affect results, lower values of urea concentration in presence of Bronopol at 0·5 g/l were observed. Urea levels in milk samples estimated by this method were comparable to standard enzymatic method. The method is simple, fast and is not prone to interference from other milk constituents.

The objective was to describe the physicochemical changes during the early phase of subclinical mastitis and to associate them with pathological findings. A Mannheimia haemolytica strain was deposited into one teat duct of 25 ewes and the clinical, bacteriological, cytological, physicochemical (pH, milk composition), gross-pathological and histological findings were subsequently recorded. The organism was consistently isolated from samples of teat duct material (140/150) but not from mammary secretion (50/150). California Mastitis Test (CMT) scores increased (>1) and remained high (143/150 samples) after challenge; polymorphonuclear neutrophils (PMN) predominated in milk films, but the proportion of lymphocytes and macrophages progressively increased. Increased pH values (>7·0) were recorded in the mammary secretion from the challenged side. Furthermore, content of fat, total proteins and lactose therein decreased markedly. Histological changes (leucocytic infiltration, destruction of epithelial cells) were observed in the mammary parenchyma of the ewes. The present results confirm that the reduction of milk constituents is the effect of cellular damage and can occur soon after infection.

Considering that several health promoting properties are associated with kefir consumption and a reliable probiotic product requires a complete identification of the bacterial species, the present work evaluates several proved markers of probiotic potential of eleven isolates of homofermentative lactobacilli isolated from kefir grains and molecular identification and genotypic diversity. Using restriction analysis of amplified ribosomal DNA (ARDRA) and analysis of the 16S–23S rRNA internal spacer region we confirmed that all homofermentative lactobacilli belong to the species Lactobacillus plantarum. RAPD-PCR analysis allowed the discrimination of lactobacilli in five clusters. All isolates exhibited high resistance to bile salt. High survival after one hour of exposure to pH 2·5 was observed in Lb. plantarum CIDCA 8313, 83210, 8327 and 8338. All isolates were hydrophilic and non autoaggregative. Isolate CIDCA 8337 showed the highest percentage of adhesion among strains. All tested lactobacilli had strong inhibitory power against Salmonella typhimurium and Escherichia coli. Seven out of eleven isolates showed inhibition against Sal. enterica and five isolates were effective against Sal. gallinarum. Only CIDCA 8323 and CIDCA 8327 were able to inhibit Sal. sonnei. We did not find any correlation between the five clusters based on RAPD-PCR and the probiotic properties, suggesting that these isolates have unique characteristics.

The somatic cell count (SCC) in milk is associated with increasing proteolytic degradation of caseins and it has been suggested that enzymes derived from somatic cells contribute to a lower yield and poorer quality of cheese. It is essential to increase the knowledge on naturally occurring milk proteinase activities to better understand how to improve the technological quality of milk. The aim of this work was to identify peptides actually present in milk as a result of proteolysis at different levels of SCC and to assign these peptides to potential responsible proteases where possible. Peptide fractions were prepared from acid whey by ultrafiltration at a molecular cut-off value of 10 000 Da. The peptides were separated using capillary reversed phase high performance liquid chromatography (RP-HPLC) and identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry (MALDI-TOF MS/MS). Peptides identified ranged in mass from 1023 to 2000 Da, and originated from αS1-, αS2- or β-casein. Possible responsible proteases that could be suggested when examining the peptide cleavage sites included plasmin, cathepsin B, D and leukocyte elastase. The results indicated that plasmin was primarily responsible for the observed proteolysis in milk at low cell count, whereas the cathepsins and elastase became implicated at elevated cell count. Specificity and activity of cathepsins and elastase has earlier mainly been studied in model systems, whereas less is known about their activities in milk itself. This is also the first indication of involvement of elastase in milk proteolysis through the unequivocal determination of cleavage sites.

Two, 8-week experiments, each using 30 lactating Holstein cows, were conducted to examine performance of animals offered combinations of total mixed ration (TMR) and high-quality pasture. Experiment 1 was initiated in mid October 2004 and Experiment 2 was initiated in late March 2005. Cows were assigned to either a 100% TMR diet (100:00, no access to pasture) or one of the following three formulated partial mixed rations (PMR) targeted at (1) 85% TMR and 15% pasture, (2) 70% TMR and 30% pasture and (3) 55% TMR and 45% pasture. Based on actual TMR and pasture intake, the dietary TMR and pasture proportions of the three PMR in Experiment 1 were 79% TMR and 21% pasture (79:21), 68% TMR and 32% pasture (68:32), and 59% TMR and 41% pasture (59:41), respectively. Corresponding proportions in Experiment 2 were 89% TMR and 11% pasture (89:11), 79% TMR and 21% pasture (79:21) and 65% TMR and 35% pasture (65:35), respectively. Reducing the proportion of TMR in the diets increased pasture consumption of cows on all PMR, but reduced total dry matter intake compared with cows on 100:00. An increase in forage from pasture increased the concentration of conjugated linoleic acids and decreased the concentration of saturated fatty acids in milk. Although milk and milk protein yields from cows grazing spring pastures (Experiment 2) increased with increasing intakes of TMR, a partial mixed ration that was composed of 41% pasture grazed in the fall (Experiment 1) resulted in a similar overall lactation performance with increased feed efficiency compared to an all-TMR ration.

The changes in milk fat globules and fat globule surface proteins of both low-preheated and high-preheated concentrated milks, which were homogenized at low or high pressure, were examined. The average fat globule size decreased with increasing homogenization pressure. The total surface protein (mg m−2) of concentrated milk increased after homogenization, the extent of the increase being dependent on the temperature and the pressure of homogenization, as well as on the preheat treatment. The concentrates obtained from high-preheated milks had higher surface protein concentration than the concentrates obtained from low-preheated milks after homogenization. Concentrated milks heat treated at 79°C either before or after homogenization had greater amounts of fat globule surface protein than concentrated milks heat treated at 50 or 65°C. This was attributed to the association of whey protein with the native MFGM (milk fat globule membrane) proteins and the adsorbed skim milk proteins. Also, at the same homogenization temperature and pressure, the amount of whey protein on the fat globule surface of the concentrated milk that was heated after homogenization was greater than that of the concentrated milk that was heated before homogenization. The amounts of the major native MFGM proteins did not change during homogenization, indicating that the skim milk proteins did not displace the native MFGM proteins but adsorbed on to the newly formed surface.

The aim of this research was to study the effects of dairy maturation on the physico-chemical characteristics and technological properties of milk used for Parmigiano-Reggiano cheese manufacture. Three different operating conditions (CF1, CF2 and CF3) were considered. Full cream milk from the evening milking was stored on the farm and delivered to the cheese factory in churns (CF1) or in thermoregulated tanks at a temperature not lower than 18°C (CF2 and CF3). The natural creaming (10–12 h overnight) was performed in a traditional large flat vat containing 10–12 hl (CF1 and CF2) or in thermoregulated large flat vats containing 60 hl at about 15°C (CF3). Twenty-four, 24 and 22 maturation trials were performed in CF1, CF2, and CF3, respectively, during 2 consecutive years. A significant increase (P⩽0·05) in pH during the maturation of milk was observed in CF1 and CF2. The increase of pH was higher (P⩽0·05) in CF1 than CF2 and CF3. The values of titratable acidity were higher (P⩽0·05) in partially skimmed evening (PS) milk than in full cream (FC) milk in each operative condition. The increase observed in CF2 was higher than those reported in CF1 and CF3. Compared with FC milk, PS milk showed lower values (P⩽0·05) of casein and casein number and higher contents (P⩽0·05) of whey proteins and, particularly, proteose-peptone. The increase of proteose-peptone – per 100 g SNF or 100 g casein – was significantly higher (P⩽0·05) in CF1 than in CF2 and, in particular, than in CF3. A higher increase (P⩽0·05) of resistance to compression was observed in CF1 with respect to CF3. CF2 variation was not different with respect to either CF1 or CF3. Variation of the difference between PS and FC milks (PS-FC) in pH, TBC and fat were clearly lower in CF3 than CF1. This means that the control of milk temperature throughout the whole maturation phase offers a greater control of both microbial development and extent of creaming.

Subclinical mastitis is a costly disease and its diagnosis is difficult. Besides the somatic cell count (SCC) and bacteriology, the differential inflammatory cell count (DICC) is a meaningful tool for mastitis detection. As microscopy is very subjective because of the low number of events to be counted, flow cytometry has often been proposed for the differentiation of milk cells. The objective of this study was to determine whether it is possible to identify subclinical mastitis in cattle at an early stage by a simple and fast flow cytometric method. The aim was to identify the main leucocyte populations in flow cytometric dotplots (polymorphonuclear neutrophils (PMN), lymphocytes and macrophages) and, with these, to elaborate a method of mastitis prognostics. Milk from 15 German Holstein cows was sampled in cross-sectional studies and SCC determined. After preparation, the milk cells were incubated with different specific antibodies that bind to different cell types and also to propidium iodide (PI), which differs between viable and non-viable cells. This procedure made it possible to localize cell types in a flow cytometric dot plot and to differentiate between viable and non-viable PMN. Percentages of viable PMN can be determined by a procedure consisting of a simple centrifugation, incubation with PI, and flow cytometric measurement. So it is possible to quickly determine the stage of the inflammation even in quarters with a low SCC.

During early lactation, many dairy cows develop fatty liver, which is associated with decreased health and reproductive performance. Currently, fatty liver can be detected reliably only by using liver biopsy followed by chemical or histological analysis, which is not practical in most on-farm situations. We tested whether digital analyses of hepatic ultrasonograms can be used to detect non-invasively fatty liver and estimate liver triacylglycerol content. A total of 49 liver biopsies and ultrasonograms were taken from 29 dairy cows within 2 weeks postpartum. The usefulness of 17 first- or second-order parameters from digital analysis of B-mode ultrasonograms were evaluated by discriminant, correlation, and regression analyses. A group of linear combinations of the 17 parameters correctly classified 40 of 49 samples into normal liver as well as mild, moderate and severe fatty liver when cut-off values were 1%, 5% and 10% and correctly classified 45 of 49 samples when cut-off values were 5% and 10% triacylglycerol of wet weight. A linear combination of 16 image parameters estimated triacylglycerol concentrations of 38 of the 39 liver samples below the cut-off value of 10% within 2·5% of liver wet weight, and a linear combination of 3 parameters estimated triacylglycerol concentrations of the 10 liver samples above the cut-off value of 10% within 2% of liver wet weight. Therefore, ultrasound imaging followed by digital analysis of sonograms has potential to non-invasively detect fatty liver and estimate liver triacylglycerol content.

There is substantial evidence showing that the polymorphism of the goat αs1-casein (CSN1S1) gene has a major effect on milk protein, casein and fat content as well as on cheese yield. However, its influence on the synthesis rate of CSN1S1 has been less studied, with measurements only available in French breeds. In this article, we have measured milk CSN1S1 content in 89 Malagueña and 138 Murciano-Granadina goats with 305 and 460 phenotypic registers, respectively. In the Malagueña breed, average values of CSN1S1 content estimated for BB, BF, EE and FF genotypes were 6·94±0·38, 5·36±0·22, 4·58±0·13 and 3·98±0·27 g/l, respectively, being all significantly different (P<0·05). Conversely, in the Murciano-Granadina breed only the BB genotype (8·50±0·60 g/l) was significantly associated with increased levels of CSN1S1 (P<0·05), whereas BF (6·56±0·82 g/l), EE (6·39±0·60 g/l) and EF (6·91±0·76 g/l) genotypes displayed non-significant differences when compared with each other. Our results highlight the existence of breed-specific genetic and/or environmental factors modulating the impact of the CSN1S1 gene polymorphism on the synthesis rate of the corresponding protein.

This study investigated the evolution of trans-9 trans-11 conjugated linoleic acid (CLA) from cis-9 trans-11 CLA during methylation and its avoidance through a rapid base methylation of milk fat. The study examined three conditions shown to result in loss of cis-9 trans-11 CLA during methylation namely: temperature, methylation time, water contamination in old reagents and acidic conditions. Three techniques currently used for the conversion of milk fat into fatty acid methyl esters for analysis of CLA content by gas liquid chromatography and a fourth procedure designed to eliminate acidic conditions and to limit methylation temperature and time were used. The four methods were: (i) acidic methylation (AM); (ii) acidic and basic bimethylation with fresh reagents (FBM); (iii) acidic and basic bimethylation with pre-prepared reagents (PBM) and (iv) basic methylation (BM). Each regime was carried out on six milk samples over two periods and methylated 1 ml freeze-dried milk (n=12 per regime). Total CLA was not different across methylation regimes (0·30 mg/ml). Isomer cis-9 trans-11 was higher (P<0·01) with BM than the other regimes and lowest with AM: 21·2, 17·8, 18·8 and 14·7 mg/100 ml for BM, FBM, PBM and AM, respectively. The inverse relationship was shown for trans-9 trans-11 with higher (P<0·001) amounts with AM than the other regimes and lowest with BM: 0·57, 2·55, 2·36 and 3·69 mg/100 ml for BM, FBM, PBM and AM, respectively. The trans-10 cis-12 isomer was also shown to alter with methylation procedure being higher (P<0·001) with AM than the other regimes: 0·43, 0·47, 0·29 and 1·20 mg/100 ml for BM, FBM, PBM and AM, respectively. Validation with known CLA free fatty acid and triacylglycerol standards confirmed that AM resulted in conversion of cis-9 trans-11 to trans-9 trans-11, and also elevated trans-10 cis-12 whilst BM of triacylglycerol CLA did not isomerise cis-9 trans-11 and was comparable to FBM.

Raw ewes' milk semi-soft cheeses (RESS-cheeses) are important products in Portugal and in several European regions. Creamy texture is an essential attribute of these cheeses, which results from structural properties that are not always well characterized. Here, the structural changes occurring during the ripening period of a traditional RESS-cheese, known as Serpa cheese, were analysed through small amplitude oscillatory shear (SAOS). Rheological data was complemented with other physical and chemical parameters, that were monitored during ripening, in order to estimate Serpa cheese ripening time using multiple linear regression (MLR). Mechanical spectra indicated a relatively strong structure, comparable to a gel, with a low dependence on frequency at the beginning of ripening and a weak structure, comparable to a concentrated suspension, with a crossing point (G″=G′) at the left of the graphic and with both moduli highly dependent on frequency, at the end of ripening. Good correlations (P<0·05) were obtained between structural (hardness and storage modulus) and proteolysis indicators. Using a combination of chemical, colour and rheological parameters we were able to obtain a multiple linear regression (MLR) which allows the estimation of Serpa cheese ripening time with an estimation error of 1·7 d (adjusted R2=0·98, P<0·0001).

Twelve spring-calving and twelve winter-calving cows were managed for extended lactation cycles of 18-months duration, with the former group then completing a second extended lactation. Half of the cows were fed according to standard management practice for the herd; the other half received supplementary concentrate from week 9 of lactation onwards. Commencing at the same time, half of the udder of each cow was subjected to increased milking frequency (thrice daily rather than twice daily). Lactation persistency (and hence total milk yield) was significantly increased by frequent milking. Winter calving cows and supplemented cows also exhibited better persistency, but this was only evident up until the point of re-breeding, at around lactation week 33. Milk composition was measured in the spring-calving cows in both their first and second extended lactations. Composition altered during the course of the lactation, protein and fat percentages increasing and lactose percentage decreasing, irrespective of treatment. The quality of the milk for processing into cheese, fermented products, heat-treated products and cream liqueurs was assessed by calculation of casein number (casein protein as a proportion of total protein). Processing quality declined across the course of lactation in those groups that showed poor persistency but not in those that maintained a persistent lactation. Milk hygienic quality (somatic cell counts) showed parallel changes. Body condition score increased during the course of lactation but was not affected by supplementation; none of the cows became excessively fat. All cows remained healthy throughout the extended lactations and the majority (33/36) re-bred successfully. By demonstrating that lactation persistency is plastic and can be improved by simple management interventions, the results lend support to the economic arguments in favour of extended lactation cycles. The likely welfare benefits of extended lactation are also discussed.

Twenty-eight Brown cows were maintained on a mountain pasture for a period of 40 days and assigned to 4 groups following a factorial design 2 stocking density (0·7 and 1·4 cows/ha)×2 supplement levels (2·4 and 4·8 kg organic matter (OM)/d). Herbage intake, animal body condition score (BCS), milk yield, milk chemical and coagulation properties, cheese composition, rheology and sensory characteristics were measured. The average herbage intake was 12·2 kg OM/d, with a significant effect related to stocking density (low, 13·1 v. high, 11·4 kg OM/d). BCS variation was always negative and changed with supplement level, although with no statistical significance (−0·43 points on average). Milk yield was lower for the group with lower availability of herbage (low supplement and high stocking density: 15·9 kg/d), whereas it was comparable among others groups (16·9 kg/d on average). The group with high supplement and low stocking density produced milk with worse cheese making properties. Cheese composition analyses showed an effect of supplement level on calcium content, and on parameters for degree of ripening, such as nitrogen fractions and lipolysis index. As for the textural parameters, hardness and gumminess were found to be higher with the low level of supplement. The different level of supplementation could differentiate the groups on the basis of the sensory perception of cheese.

The objective was to compare the relationships between luteal activity and fertility, and relate these parameters to metabolic indices and body condition changes in multiparous Estonian Holstein cows on two commercial dairy farms under different management and levels of production and nutrition (higher, H, n=54 (71 lactations) and lower, L, n=39 (39 lactations)). For statistical analysis cows were categorized according to their milk progesterone (P4) profiles as follows: normal ovarian function; delayed start of cyclicity (DC) (interval from calving to first luteal response (P4⩾5 ng/ml up to and more than 50 d respectively, followed by regular cyclicity); cessation of luteal activity (prolonged interluteal interval, P4<5 ng/ml, with a duration of ⩾14 d between two adjacent luteal phases); prolonged luteal activity (P4 levels ⩾5 ng/ml for ⩾20 d without preceding insemination). The Mixed procedure of the SAS system was used to compare milk production traits, blood metabolites (ketone bodies, non-esterified fatty acids, total cholesterol) and aspartate aminotransferase, body condition scores (BCS) and fertility parameters between the two farms, and also fertility parameters between the farms within P4 categories. Differences in milk fat/protein ratio, ketone body levels and BCS indicated a deeper negative energy balance (NEB) during the first month after calving on farm L. On both farms nearly 50% of the recently calved dairy cows suffered from ovarian dysfunction during the post-partum period. Delayed start of cyclicity was the most prevalent abnormal P4 profile, 25% and 28% on farms H and L, respectively. Prolonged luteal activity accounted for one-third of atypical ovarian patterns on farm H, and cessation of luteal activity on farm L. On farm L, DC cows had lower BCS values from day 10 to day 90 after calving compared with normal cows (P<0·01) and cows lost more BCS (1·2 units) during the 40 d after calving than normal resumption cows (0·75 units; P<0·05). On farm H with moderate NEB the delayed start of ovulation post partum did not impair subsequent reproductive performance.