In dairy cows the period of transition from late gestation to early lactation is recognized as inducing considerable metabolic adaptation. The aim of this study was to analyse modifications in serum protein values occurring during the dry and the transition period and during lactation in a group of five Holstein cows of high average milk production. For all subjects, selected on the basis of their pregnancy status, blood samples were collected at different physiological phases: dry period (−60, −30 d to calving), transition period (almost 7 d to calving, 7 d after calving), and lactation (weeks 2, 5 and 15 after calving), for a total of eight blood samples for each cow. On each blood sample total proteins and electrophoresis analysis were performed. On the data obtained, normally distributed (P<0·05, Kolmogorov-Smirnov's test), one-way Repeated Measure Analysis of Variance (ANOVA), was applied to evaluate the influence of different stages of gestation and lactation on the considered parameters. Results showed a significant effect on total proteins, α1-globulins, β-globulins, γ-globulins and albumin/globulin ratio. Most of the detected modifications were related to the transition from gestation to lactation, indicating that it is a period of great metabolic stress for cows. On the basis of the obtained results we can affirm that the pattern of serum protein fraction rn could give information about dehydration, plasma volume expansion and hepatic function occurring during the peripartum period in dairy cows.

Flax hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on digestion of flax hull based diets and nutritive value of flax hull for dairy production. Flax oil is rich in α-linolenic acid (LNA) and rumen bypass of flax oil contributes to increase n-3 FA proportions in milk. Therefore, the main objective of the experiment was to determine the effects of abomasal infusion of increasing amounts of flax oil on apparent digestibility, dry matter (DM) intake, milk production, milk composition, and milk FA profile with emphasis on the proportion of LNA when cows were supplemented or not with another source of LNA such as flax hull. Six multiparous Holstein cows averaging 650±36 kg body weight and 95±20 d in milk were assigned to a 6×6 Latin square design (21-d experimental periods) with a 2×3 factorial arrangement of treatments. Treatments were: 1) control, neither flax hull nor flax oil (CON), 2) diet containing (DM basis) 15·9% flaxseed hull (FHU); 3) CON with abomasal infusion of 250 g/d flax oil; 4) CON with abomasal infusion of 500 g/d flax oil; 5) FHU with abomasal infusion of 250 g/d flax oil; 6) FHU with abomasal infusion of 500 g/d flax oil. Infusion of flax oil in the abomasum resulted in a more pronounce decrease in DM intake for cows fed the CON diets than for those fed the FHU diets. Abomasal infusion of flax oil had little effect on digestibility and FHU supplementation increased digestibility of DM and crude protein. Milk yield was not changed by abomasal infusion of flax oil where it was decreased with FHU supplementation. Cows fed FHU had higher proportions of 18:0, cis9-18:1, trans dienes, trans monoenes and total trans in milk fat than those fed CON. Proportion of LNA was similar in milk fat of cows infused with 250 and 500 g/d flax oil in the abomasum. Independently of the basal diet, abomasal infusion of flax oil resulted in the lowest n-6:n-3 FA ratio in milk fat, suggesting that the most important factor for modification of milk FA profile was the amount of n-3 FA bypassing the rumen and not the amount of flax hull fed to dairy cows. Moreover, these data suggest that there is no advantage to supply more than 250 g/d of flax oil in the abomasum to increase the proportion of LNA in milk fat.

Bacteriological analysis of milk samples from quarters of a dairy cow suffering from subclinical mastitis yielded two isolates of Staphylococcus aureus which gave a negative reaction in the standard coagulase test. Both isolates were also clumping factor and thermonuclease negative, and gave a negative reaction in the Staphaurex® test. The isolates were identified by using commercial biochemical systems, and by PCR analysis of different staphylococcal cell surface protein and exoprotein genes. Further molecular identification of the isolates, which included sequencing of the 16S rRNA gene and RT-PCR of coagulase (coa), clumping-factor (clfA) and thermonuclease (nuc) genes, was consistent with the diagnosis phenotypically ‘coagulase-negative variant of Staph. aureus’. The fact that coagulase-negative Staph. aureus variants can occur in the context of intramammary infections in cattle may result in the incorrect diagnosis ‘coagulase-negative staphylococci (CNS)’ in routine mastitis diagnostic, at least in rare cases. To fully ensure correct species diagnosis, sequencing of the 16S rRNA gene and amplification of specific genes such as coa is necessary in these cases.

The present study investigated the effect of age at first lambing (AFL) on the performance of Lacaune sheep under intensive management conditions. Records from 3088 maiden sheep from one farm, for the period 2005–2010, were classified into four experimental groups: group E (early) ewes with AFL ⩽390 d; group M (middle) with AFL of 391–450 d; group L (late) with AFL of 451–510 d; and group A (aged) with AFL ⩾511 d. The higher the number of lactations, the lower were the yield/lactation and yield/Day in Milk. Ewes from group M up to 450 days old lambed 0·2 times more often and had 0·25 more lactations than the ewes from group L; in addition, the former group lambed 0·5 times more often and had 0·49 more lactations than ewes from group A. Group A had the lowest yield per lifetime; the yield followed the sequence: group E (1032 l) >group M (1051 l) >group L (989 l) > group A (859 l) (P<0·0001). Yield/lactation was affected by AFL during the first three lactations (P<0·05). Group E produced significantly less milk in the first lactation than ewes from groups M, L and A. AFL correlated negatively with the number of lactations in life (r= −0·26; P<0·0001) and with total milk yield (r= −209; P<0·0001). The productive performance between the ewes that reached puberty early (n=404) or late (n=2684) in life were different (P<0·0001) with the early sheep having a lower total milk yield (591 v. 1073 l, P<0·0001). In conclusion, the current study indicates that the optimal AFL for the Lacaune breed under an intensive dairy system is between 390 and 450 d. Ewes with AFL earlier than 390 d or later than 450 d are likely to have a shorter productive life and a lower lifetime milk production.

This study was undertaken to compare flow cytometry (FC) and direct microscopic leucocyte count (MDLC) for the differentiation of macrophages, lymphocytes and polymorphonuclear leucocyte (PMN) and to evaluate leucocyte distribution in ewe milk with low and high somatic cell count (SCC). Milk samples were grouped for somatic cell count in low SCC (LSCC) when the content was lower than 5·00×105/ml and high SCC (HSCC) when the content was higher than 1·00×106/ml. No differences were found between the two methods tested suggesting that FC could be used as a routine test for rapid discrimination of leucocytes. Percentages of lymphocytes in ewe milk were higher in LSCC (50%) than in HSCC (39%) and count ranged from 273·91±56·62×103cells/ml (LSCC) to 308·90±46·15×103cells/ml (HSCC). PMN number was lower in LSCC than in HSCC (248·83±46·87×103cells/ml v. 444·38±58·62×103cells/ml); accordingly the percentage was lower in LSCC (40%) than in HSCC (57%). No differences were found for macrophages which were 36·36±5·51×103cells/ml and 39·32±6·83×103cells/ml in LSCC and HSCC, respectively. Lymphocytes in ewe milk did not vary with increased number of somatic cells and were the predominant cell type in LSCC. PMN represented the main population detected in HSCC and the correlation with SCC evidenced that this leucocyte class could be useful in differentiating ewe milk cell count, being strictly responsible for the SCC increase.

Somatic cell count (SCC) is a widely used marker of udder health and a predictor of inflammation caused by an immune response. The objective of this study was to determine whether selected measures of mammary gland health as well as milk fatty acid profile were altered by an increase in milking frequency using a unilateral frequent milking (UFM) model. Holstein cows at parturition were assigned to UFM, in which the left udder half of each cow was milked four-times daily (4X) and the right udder half was milked twice daily (2X) for the first 21 days in milk (DIM). Milk yields from each udder half were measured from 1–21 DIM and samples were collected on days 3, 7, 10, 14 and 21 for determination of SCC and milk composition. Flow cytometric analysis with bovine monoclonal antibodies was used to identify milk immune cell populations and milk fatty acid (FA) composition was determined using gas chromatography. Gene expression analysis was used to determine whether there was an alteration in mRNA expression of genes involved in milk fat production including lipoprotein lipase (LPL) and FA-binding protein 3 (FABP3) with ribosomal protein S9 (RPS9) as a house-keeping gene. No difference was detected for milk SCC or cell populations between the udder halves milked 4X as compared with the udder halves milked 2X. In addition, no difference was detected for any FA in milk from the udder half milked 4X as compared with the udder half milked 2X. Overall, using a UFM model, increased milking frequency for the first 21 DIM did not affect selected measures of mammary gland health or milk FA, but was associated with greater milk yield, milk fat percent and yield, and milk protein and lactose yields.

Feeding agro-industrial by-products and unconventional forages, rich in potentially anti-nutritional factors, may influence the quality of the raw milk and the dairy products prepared therefrom. The aim of the present study was to determine side-effects on physicochemical properties of milk, yoghurt and cheese of feeding diets where one third were feeds either rich in lipids (tomato pomace and olive cake) or phenols (olive leaves and lentil straw) or electrolytes (Atriplex leaves). The diets, including a control diet, were designed to be isoenergetic and isonitrogenous. They were fed in amounts of 2 5 kg dry matter/day per head during 50 days to 6×10 multiparous fat-tailed Awassi ewes. Milk samples were analysed for various physicochemical traits and fatty acid composition on days 0, 24, 36 and 48. Three times, milk pooled by group was processed to yoghurt and non-aged farmer-type cheese, which were analysed for their gross and fatty acid composition and texture, and were subjected to sensory evaluation. Feeding olive cake and tomato pomace reduced milk casein, but increased proportions of monounsaturated fatty acids. There were some variations in minerals among test diets but, contrary to expectations, Atriplex did not increase milk sodium. The nutritional composition of yoghurt and cheese was not varied much by the test feeds, except for some changes in fatty acid profile similar to the milk. Yoghurt firmness declined with all test diets, but texture score tended to be lower only for olive cake and leaf diets relative to control. Cheese firmness was increased by feeding the Atriplex leaf and olive cake diets which was also reflected in the texture scores. No off-flavours were reported. Possible reasons for effects on the dairy products are discussed. In conclusion, the feeds investigated had certain effects on the physicochemical properties of dairy products, but these were neither very systematic nor large thus not prohibiting their use in Mediterranean sheep milk production systems.

The objective of the study was to evaluate the relationship of veterinary clinical assessments of lameness to probability estimates of lameness predicted from vertical kinetic measures. We hypothesized that algorithm-derived probability estimates of lameness would accurately reflect vertical measures in lame limbs even though vertical changes may not inevitably occur in all lameness. Kinetic data were collected from sound (n=179) and unilaterally lame (n=167) dairy cattle with a 1-dimensional, parallel force plate system that registered vertical ground reaction force signatures of all four limbs as cows freely exited the milking parlour. Locomotion was scored for each hind limb using a 1–5 locomotion score system (1=sound, 5=severely lame). Pain response in the interdigital space was quantified with an algometer and pain response in the claw was quantified with a hoof tester fitted with a pressure gage. Lesions were assigned severity scores (1=minimal pathology to 5=severe pathology). Lameness diminished the magnitude of peak ground reaction forces, average ground reaction forces, Fourier transformed ground reaction forces, stance times and vertical impulses in the lame limbs of unilaterally lame cows. The only effect of lameness on the opposite sound limb was increased magnitude of stance times and vertical impulses in unilaterally lame cows. Symmetry measures of the peak ground reaction forces, average ground reaction forces, Fourier transformed ground reaction forces, stance times and vertical impulses between the left and right hind limbs were also affected in unilateral lameness. Paradoxically, limbs with clinically similar lesion and locomotion scores and pain responses were associated with a broad range of load-transfer off the limb. Substantial unloading and changes in the vertical limb variables occurred in some lameness while minimal unloading and changes in vertical limb variables occurred in other lameness. Corresponding probability estimates of lameness accurately reflected changes in the vertical parameters of limbs and generated low probability estimates of lameness when minimal unloading occurred. Failure to transfer load off limbs with pain reactions, locomotion abnormalities and lesions explained much of the limited sensitivity in lameness detection with vertical limb variables.

Up to half of quarter milk samples submitted for mastitis diagnosis are culture-negative results or lead to identification of coagulase-negative staphylococci or Corynebacterium bovis in conventional culturing, the so-called minor pathogens. The interpretation and usefulness of these results in terms of udder and animal health management is limited, even though the amount of resources spent is relatively high. This work aimed to test two methods of analysis of milk samples with the goal of increasing detection of intramammary pathogens. In the first study, 783 milk samples were processed in duplicate: before and after freezing at −20°C for 24 h, using standard bacteriological techniques. There was a significant difference between the two methods with samples frozen for 24 h yielding significantly fewer Gram-positive catalase-positive cocci, Gram-negative bacilli, Gram-positive bacilli and significantly more samples leading to no growth, than samples before freezing. The number of samples yielding Gram-positive catalase-negative cocci was not significantly affected by freezing. In the second study, a real-time PCR-based test was performed on milk samples with an individual quarter somatic cell count above 500 000 cells/ml that were either negative (n=51 samples) or that led to the isolation of minor pathogens in culturing: Corynebacterium bovis (n=79 samples) or non-aureus staphylococci (NAS, n=32). A mastitis pathogen, beyond the result obtained with standard bacteriology, was detected on 47% of the no-growth samples, on 35% of the samples from which C. bovis had been isolated and on 25% of the samples from which NAS had been isolated. The most commonly detected major pathogen was Escherichia coli, followed by Streptococcus uberis, Arcanobacterium pyogenes/Peptoniphilus indolicus and Streptococcus dysgalactiae. These results suggest that simply freezing milk samples for 24 h does not increase the detection of intramammary bacteria in milk samples and therefore should not be recommended. However, use of the real-time PCR-based test may be useful in diagnosing intramammary infections when milk samples with high somatic cell counts are culture-negative or when culturing results in the detection of minor pathogens.

The objectives of this study were to analyse buffalo milk fat composition, to verify the activity of Delta(9)-desaturase enzyme in the mammary gland, as well as to estimate additive genetic variances for milk, fat and protein yield, and milk cis-9,trans-11 conjugated linoleic acid percentage (cis-9,trans-11 CLA%). A total of 3929 lactation milk yields (MY) records from 2130 buffaloes and 1598 lactation fat (FY) and protein (PY) yield records from 914 buffaloes were analysed. For cis-9,trans-11 CLA%percentage, a total of 661 milk samples from 225 buffaloes, daughters of 8 sires, belonging to 4 herds and calving in 2003 and 2004, were used. The genetic parameters and variance components were estimated by Restricted Maximum Likelihood applying an animal model. The fixed effects considered in the model were: contemporary group (herd, year, calving season) and age at calving (linear and quadratic effects) and lactation length (linear and quadratic effects) as covariables. Additive genetic and permanent environment effects were considered as random. The MY, FY, PY and CLA% means were 1482±355 kg, 90·1±24·6 kg, 56·9±15·2 kg and 0·69±0·16%, respectively. Heritability estimates for MY, FY, PY and CLA% were 0·28±0·05, 0·26±0·11, 0·25±0·11 and 0·35±0·14, respectively. There is enough additive genetic variation for buffalo milk, protein and fat yield to improve these traits through selection. The cis-9,trans-11 CLA% can be enhanced by selection in buffaloes and will contribute to improving human health. The activity and efficiency of Delta(9)-desaturase in the mammary was measured and confirmed.

The aim of the study was to investigate the effects of season, cow cleanliness and milking routine on bacterial and somatic cell counts of bulk tank milk. A total of 22 dairy farms in Lombardy (Italy) were visited three times in a year in different seasons. During each visit, samples of bulk tank milk were taken for bacterial and somatic cell counts; swabs from the teat surface of a group of cows were collected after teat cleaning and before milking. Cow cleanliness was assessed by scoring udder, flanks and legs of all milking cows using a 4-point scale system. Season affected cow cleanliness with a significantly higher percentage of non-clean (NC) cows during Cold compared with Mild season. Standard plate count (SPC), laboratory pasteurization count (LPC), coliform count (CC) and somatic cell count, expressed as linear score (LS), in milk significantly increased in Hot compared with Cold season. Coagulase-positive staphylococci on teat swabs showed higher counts in Cold season in comparison with the other ones. The effect of cow cleanliness was significant for SPC, psychrotrophic bacterial count (PBC), CC and Escherichia coli in bulk tank milk. Somatic cell count showed a relationship with udder hygiene score. Milking operation routine strongly affected bacterial counts and LS of bulk tank milk: farms that accomplished a comprehensive milking scheme including two or more operations among forestripping, pre-dipping and post-dipping had lower teat contamination and lower milk SPC, PBC, LPC, CC and LS than farms that did not carry out any operation.

Conjugated linoleic acid (CLA) reduces mammary milk fat synthesis in a dose-dependent manner. Our objective was to determine the effects of lipid-encapsulated CLA (LE-CLA) supplementation on milk production, reproductive performance and metabolic responses in lactating dairy cows fed a grass silage-based diet. Seventy-two Holstein-Friesian cows (32 primiparous and 40 multiparous) were used in a completely randomized block design. Cows received either 80 g of LE-CLA daily or 60 g of calcium salts of palm fatty acids daily (control) from parturition until 60 days in milk. LE-CLA contained a 50:50 mix of cis-9,trans-11 CLA and trans-10,cis-12 CLA, resulting in a daily intake of 6 g of each isomer. Milk production and dry matter intake were recorded daily, and blood samples were collected 3-times a week. Blood samples were analysed for circulating concentrations of glucose, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), insulin and insulin-like growth factor-I (IGF-I). Progesterone was measured in blood samples collected after the first post-partum insemination. Ovarian ultrasound examinations commenced at 8–10 d post partum and were carried out 3-times a week until first ovulation. LE-CLA treatment resulted in decreased milk fat concentration, with consequent improvements in energy balance and body condition score (BCS). The peak concentration of NEFA in blood was reduced by LE-CLA, but circulating concentrations of insulin, glucose, IGF-I, BHBA and progesterone were not affected. There was no effect of LE-CLA supplementation on the post-partum interval to first ovulation. Services per conception tended to be reduced. The reduction in milk energy output and improvement in energy status and BCS in LE-CLA-supplemented cows provides a strong rationale for further studies with greater cow numbers to test effects on reproductive performance.

Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria from milk samples in several studies worldwide. Despite their relative frequency, specific measures aiming at their control are not well established. One possible measure to include in a control programme is lactational antimicrobial treatment. The decision to perform such treatment, as well as other actions on farm, should be based on the likelihood of financial return. A deterministic model was used to evaluate whether performing an antimicrobial treatment during the lactation for quarters infected with CNS was financially justifiable. Input variables for the impact of CNS on udder health were based on a previous study by the same authors and on available literature on the subject. Prices included in the model were based on 2009/2010 conditions in Portugal. The average result per antimicrobial treated quarter was a net loss of €38·74. Performing a sensitivity analysis to evaluate how systematic variation of the input variables of the model would lead to outcome changes showed that variation in input variables nearly always led to a negative outcome, with the greatest variation in losses observed for variation in the length of treatment and milk withdrawal period (−€46·26 to −€28·49). The situations in which a net benefit was to be expected included the bulk tank somatic cell count decreasing to a level corresponding to a premium payment or to penalties being avoided, and the prevention of transmission of CNS in the milking parlour when the possibility of transmission was at its highest level. For most situations, lactational treatment of CNS subclinical mastitis was not financially justifiable.

Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on its value for dairy production. Monensin supplementation is known to modify biohydrogenation of FA by rumen microbes. Therefore, the main objective of the experiment was to determine the effect of feeding a combination of monensin and flaxseed hulls on ruminal fermentation characteristics and FA profile of ruminal fluid and milk. Four ruminally fistulated multiparous Holstein cows averaging 665±21 kg body weight and 190±5 d in milk were assigned to a 4×4 Latin square design (28-d experimental periods) with a 2×2 factorial arrangement of treatments. Treatments were: 1) control, neither flaxseed hulls nor monensin; 2) diet containing (dry matter basis) 19·8% flaxseed hulls; 3) diet with monensin (16 mg/kg dry matter); 4) diet containing 19·8% (dry matter basis) flaxseed hulls and 16 mg monensin/kg. Flaxseed hull supplementation decreased the acetate to propionate ratio in ruminal fluid and monensin had no effect. Concentrations of trans-18:1 isomers (trans9,trans11,trans13/14+6/8) and cis9,12,15-18:3 in ruminal fluid and milk fat were higher and those of cis9,12-18:2 in milk fat tended (P=0·07) to be higher for cows supplemented with flaxseed hulls than for cows fed no flaxseed hulls. Monensin had little effect on milk fatty acid profile. A combination of flaxseed hulls and monensin did not result in better milk fatty acid profile than when feeding only flaxseed hulls.

This study was undertaken to assess the effect of somatic cell count in ewe milk on i) composition and hygienic traits; ii) plasmin, cathepsin and elastase activities; iii) leukocyte differential count; iv) renneting parameters. Individual ewe milk samples were grouped according to somatic cell count (SCC) into five classes: SC300 (<300 000 cells/ml), SC500 (from 301 000 to 500 000 cells/ml), SC1000 (from 501 000 to 1 000 000 cells/ml), SC2000 (from 1 001 000 to 2 000 000 cells/ml) and SC>2000 (>2 001 000 cells/ml). Individual milk samples were analysed for pH, chemical composition, microbial features, indigenous proteolytic enzymes, differential leukocyte population, and renneting parameters. Milk yield, lactose, protein, non casein nitrogen, microbial features were affected by SCC level. Plasmin and elastase activities were the highest in samples with more than 1 000 000 cells/ml; plasmin had intermediate values in samples with 300 000 to 1 000 000 cells/ml and the lowest in samples with less than 300 000 cells/ml of milk. Cathepsin D showed significantly lower values in SC300 and SC1000 classes than in SC500, SC2000 and SC>2000 classes. The highest percentages of lymphocyte were found in samples with less than 1 000 000 cells/ml, while the highest levels of polymorphonuclear leukocyte were found in samples with more than 1 000 000 cells/ml of milk. Longer clotting time was found in SC>2000 samples, while reduced clot firmness was observed in SC500 and SC>2000 samples. Results on milk yield and on compositional parameters evidenced an impairment of udder efficiency in ewe milk samples starting from 300 000 cells/ml. Plasmin activity in milk can be considered as a marker of the synthetic and secreting ability of the mammary gland; furthermore plasmin and elastase were consistent with the health status of the udder. Finally cathepsin D played a role in the worsening of renneting properties of ewe milk.

Sensitive methods that are currently used to monitor proteolysis by plasmin in milk are limited due to their high cost and lack of standardisation for quality assurance in the various dairy laboratories. In this study, four methods, trinitrobenzene sulphonic acid (TNBS), reverse phase high pressure liquid chromatography (RP-HPLC), gel electrophoresis and fluorescamine, were selected to assess their suitability for the detection of proteolysis in milk by plasmin. Commercial UHT milk was incubated with plasmin at 37°C for one week. Clarification was achieved by isoelectric precipitation (pH 4·6 soluble extracts) or 6% (final concentration) trichloroacetic acid (TCA). The pH 4·6 and 6% TCA soluble extracts of milk showed high correlations (R2 > 0·93) by the TNBS, fluorescamine and RP-HPLC methods, confirming increased proteolysis during storage. For gel electrophoresis, extensive proteolysis was confirmed by the disappearance of α- and β-casein bands on the seventh day, which was more evident in the highest plasmin concentration. This was accompanied by the appearance of α- and β-casein proteolysis products with higher intensities than on previous days, implying that more products had been formed as a result of casein breakdown. The fluorescamine method had a lower detection limit compared with the other methods, whereas gel electrophoresis was the best qualitative method for monitoring β-casein proteolysis products. Although HPLC was the most sensitive, the TNBS method is recommended for use in routine laboratory analysis on the basis of its accuracy, reliability and simplicity.

Standard method for the determination of the activity of milk coagulants is the rotating bottle method, where clotting time is defined as the time when visually observable flocculation starts. Aim of this study was to verify whether it is possible to determine milk clotting time by rotational viscometry. Using three different coagulants and reconstituted milk of different pH and temperature, flocculation time and viscosity in steady shear was determined, and milk gelation was monitored by small amplitude oscillating shear rheometry. The results show that, independent of pH and temperature, milk clotting time is related to an apparent viscosity of 7·24±0·45 mPa.s, indicating that rotational viscometry can be used for the screening of flocculation time with an accuracy of approximately 6%.

This study was designed to evaluate the effects of a single intramammary infusion of Panax ginseng extract on cell proliferation and death mechanism in bovine mammary gland during early involution. Eight mammary quarters from six non-pregnant cows in late lactation were infused with 10 ml of ginseng solution (3 mg/ml), six quarters were treated with 10 ml of placebo (vehicle alone) and six quarters were maintained as uninoculated controls. Milking was interrupted after infusion. Animals included in the three groups were slaughtered 7 d after inoculation and samples for histological analysis were taken. Morphometric analysis showed a significant increase in percentages of mammary tissue area occupied by stroma in ginseng-treated quarters compared with controls. A significant increase of immunostained area for bax protein and active caspase-3 was observed in ginseng-treated quarters compared with controls, whereas no differences were observed for bcl-2 immunostaining. Expression of bax mRNA was significantly higher in ginseng-treated quarters than in controls. The bax/bcl-2 ratio indicated a significant predominance of bax over bcl-2 mRNA expression in ginseng-treated quarters compared with controls. The rise of epithelial and stromal cell apoptosis in situ by TUNEL was more marked in quarters treated with ginseng than in controls. Ginseng inoculation had no effect on the number of epithelial and stromal proliferating cells labelled with Ki-67 antibody. Ratio of apoptotic to proliferating cells was higher in quarters treated with ginseng compared with controls, indicating a net loss of cells in parenchymal components. Also, the intramammary inoculation of ginseng extract at drying off increased the rate of mammary cell apoptosis without inhibiting cell proliferation. Taken together, these changes are indicative of mammary regression enhancement during early involution.

The study of bovine mammary gland functional genomics requires appropriate cDNA library collections to access gene expression patterns from different developmental and physiological stages. The present study was undertaken with the objective to identify candidate genes involved in the process of increased milk synthesis following 0, 48 and 96 h of recombinant bovine somatotropin (rbST) treatment to Surti buffalo (Bubalus bubalis) through differential display reverse transcriptase PCR (DDRT-PCR). Of a total 50 sequenced DD bands, 64% of ESTs were differentially expressed (appeared only in post-treatment samples, i.e. 48 h and 96 h) and 36% were up-regulated after rbST treatment. Of the ESTs 32%were found to be located on Bos taurus chromosome 24 (equivalent to buffalo chromosome 22), whereas 16% of ESTs could not be mapped, indicating that they are specific to buffalo. Quantitative real time PCR assay of 15 ESTs revealed transcript level surge in 13 ESTs, and decline in one EST, while one showed up-regulation in expression level at 48 h while down-regulation at 96 h. This study indicates more than 30 novel transcripts, with unknown function, involved in increased milk synthesis and also the involvement of many more genes in the physiology of milk production than once thought.

Somatic cell count (SCC) is generally regarded as an indicator of udder health. A cut-off value of 100×103 cells/ml is currently used in Germany to differentiate between normal and abnormal secretion of quarters. In addition to SCC, differential cell counts (DCC) can be applied for a more detailed analysis of the udder health status. The aim of this study was to differentiate somatic cells in foremilk samples of udder quarters classified as normal secreting by SCC <100×103 cells/ml. Twenty cows were selected and 72 normal secreting udder quarters were compared with a control group of six diseased quarters (SCC >100×103 cells/ml). In two severely diseased quarters of the control group (SCC of 967×103 cells/ml and 1824×103 cells/ml) Escherichia coli and Staphylococcus aureus were detected. DCC patterns of milk samples (n=25) with very low SCC values of ⩽6·25×103 cells/ml revealed high lymphocyte proportions of up to 92%. Milk cell populations in samples (n=41) with SCC values of (>6·25 to ⩽25)×103 cells/ml were also dominated by lymphocytes (mean value 47%), whereas DCC patterns of milk from udder quarters (n=6) with SCC values (>25 to ⩽100)×103 cells/ml changed. While in samples (n=3) with SCC values of (27–33)×103 cells/ml macrophages were predominant (35–40%), three milk samples with (43–45)×103 cells/ml indicated already inflammatory reactions based on the predominance of polymorphonuclear leucocytes (PMN) (54–63%). In milk samples of diseased quarters PMN were categorically found as dominant cell population with proportions of ⩾65%. Macrophages were the second predominant cell population in almost all samples tested in relationship to lymphocytes and PMN. To our knowledge, this is the first study evaluating cell populations in low SCC milk in detail. Udder quarters classified as normal secreting by SCC <100×103 cells/ml revealed already inflammatory processes based on DCC.