Optimising the number of replacement heifers needed will have positive economic and environmental consequences on herds that rear their own young stock. The number of heifers needed to be kept is closely related with the number of culled dairy cows in the herd. This study therefore looked at the variation that exists in culling rate and herd level factors associated with it. A dataset from 1903 dairy herds available included information at animal level (dates of culling, slaughter/death) and herd level (characteristics of reproduction, performance, health) over the years 2007 to 2010. The average culling rate for slaughter/death was used and was defined for each year as percentage of the herd size that died within 30 d after they were culled. The analysis of the association between average culling rate for slaughter/death and the characteristics of the herd was performed using a mixed model. The results showed that the average culling rate for slaughter/death was 25·4% and varied between 23% (2007) and 28% (2010). More than 70% of the herds have an average culling rate for slaughter/death of less than 30%, showing that there is room for lowering the average culling rate for slaughter/death. A higher average culling rate for slaughter/death is associated with a longer average calving interval, a higher average 305-d protein production, a higher average somatic cell count (SCC), a higher percentage of new high SCC, a more than 5% decrease in herd size, and herds that bought more than 1% of animals per year. A lower average culling rate for slaughter/death is associated with a longer average age, herds that bought less than 1% of animals per year and a more than 5% increase in herd size. In conclusion, the average culling rate for slaughter/death is associated with fertility, udder health and openness of the herd.

The objective of the present paper is to review the significance of administration of antibiotics at the end of a lactation period/beginning of the dry-period in ewes. During the stage of active involution, there is an increased risk of new mastitis cases and recrudescence of subclinical infections that had occurred during the previous lactation period. The main pathogens involved in the so-called ‘dry-period mastitis’ are coagulase-negative staphylococci. The principle of antibiotic administration at the end of a lactation period involves the intramammary infusion of a preparation to both mammary glands of ewes in the flock. Although a variety of products is licensed for administration in ewes, preferably the product for administration should be selected on the results of susceptibility testing of bacteria to be isolated from samples from ewes in the flock. In many clinical studies from around the world, performed in dairy- or mutton-production flocks, administration of antimicrobial agents at the end of a lactation period has been found beneficial in curing intramammary infections present at cessation of a lactation period, as well as in minimising the risk for intramammary infections during the dry-period. In dairy flocks, there are also benefits from increase in milk yield and decrease flock bulk milk mean somatic cell counts during the subsequent lactation period. Antibiotic administration at drying-off may be performed to all animals in a flock (‘complete’) or only to those considered to be infected (‘selective’). In all cases, after administration of the antibiotic, definite and complete cessation of the lactation period is essential for success of the procedure. Moreover, maintenance of the prescribed withdrawal periods is essential to safeguard public health. The procedure should always be applied as part of a strategic udder health management plan in a flock; implementation improves the welfare of animals and affords significant financial benefits to the farmer. A mastitis prevention scheme during lactation will minimise the incidence of the disease; effective treatment of cases of the disease during lactation will decrease the bacterial populations in the flock and limit risk of infection of other animals. Administration of antibiotics at the end of a lactation period will complement the above procedures and will contribute to improved mammary health for the forthcoming lactation period.

We investigated the mucus-binding properties of aggregating and non-aggregating potentially probiotic strains of kefir-isolated Lactobacillus kefiri, using different substrates. All the strains were able to adhere to commercial gastric mucin (MUCIN) and extracted mucus from small intestine (SIM) and colon (CM). The extraction of surface proteins from bacteria using LiCl or NaOH significantly reduced the adhesion of three selected strains (CIDCA 8348, CIDCA 83115 and JCM 5818); although a significant proportion (up to 50%) of S-layer proteins were not completely eliminated after treatments. The surface (S-layer) protein extracts from all the strains of Lb. kefiri were capable of binding to MUCIN, SIM or CM, and no differences were observed among them. The addition of their own surface protein extract increased adhesion of CIDCA 8348 and 83115 to MUCIN and SIM, meanwhile no changes in adhesion were observed for JCM 5818. None of the seven sugars tested had the ability to inhibit the adhesion of whole bacteria to the three mucus extracts. Noteworthy, the degree of bacterial adhesion reached in the presence of their own surface protein (S-layer) extract decreased to basal levels in the presence of some sugars, suggesting an interaction between the added sugar and the surface proteins. In conclusion, the ability of these food-isolated bacteria to adhere to gastrointestinal mucus becomes an essential issue regarding the biotechnological potentiality of Lb. kefiri for the food industry.

On two farms, three milking groups of buffalo cows were used to assess the consistency of entrance order and the preference for one side of the milking parlour. On Farm 1 (F1) all animals were primiparous (n=57). On Farm 2, three primiparous, 16 secondiparous and 36 multiparous cows (range 1–8) constituted group F2G1; whereas group F2G2 had 12 primiparous, 10 secondiparous and 14 multiparous cows (range 1–10). Animals were milked in auto-tandem milking parlours (2×5 and 2×6 for Farms 1 and 2, respectively). For each cow, entrance order into milking parlour, side where she was milked, milk yield, time and duration of milking were recorded. These data were derived from the computerised identification of cows. The sequence in which the cows entered the milking parlour ranged from 1 to 57 for group F1, from 1 to 55 for group F2G1 and from 1 to 36 for group F2G2. The analysis of data was conducted on 130, 120 and 92 consecutive milkings for groups F1, F2G1 and F2G2, respectively. Kendall's coefficients of concordance showed a strong constancy of the entrance order into milking parlour for groups F1 (W=0·658; χ2=4792·81; P<0·001), F2G1 (W=0·779; χ2=5046·81; P<0·001) and F2G2 (W=0·624; χ2=2030·48; P<0·001). Spearman rank correlation coefficients indicated that the more productive cows in groups F1 and F2G1 tended to enter the milking parlour first (rs=−0·221 and rs=−0·215; P<0·10; respectively). In group F2G1, a negative correlation was found between duration of milking and order of entry in the milking parlour (rs=−0·265; P<0·05). Animals in group F2G2 (rs=0·334; P<0·05) with higher days in milk entered the milking parlour latterly. In all three groups, 68 cows (45·9%) preferred the right side of the milking parlour, 73 the left side (49·3%) and the remaining seven (4·8%) showed no preference. Finally, negative correlations were found between mean entrance order and parity for both groups of Farm 2 (rs=−0·319; P<0·05 and rs=−0·325; P<0·05 for F2G1 and F2G2, respectively). As buffaloes showed higher entrance order consistency and side preference than other domestic ruminants, it is concluded that management practices that disturb their choice should be avoided in order to minimise stress during farming routines.

Lactoferrin and lysozyme are antimicrobial and immunomodulatory proteins produced in high quantities in human milk that aid in gastrointestinal (GI) health and have beneficial effects when supplemented separately and in conjunction in human and animal diets. Ruminants produce low levels of lactoferrin and lysozyme; however, there are genetically engineered cattle and goats that respectively secrete recombinant human lactoferrin (rhLF-milk), and human lysozyme (hLZ-milk) in their milk. Effects of consumption of rhLF-milk, hLZ-milk and a combination of rhLF-and hLZ-milk were tested on young pigs as an animal model for the GI tract of children. Compared with control milk-fed pigs, pigs fed a combination of rhLF and hLZ (rhLF+hLZ) milk had a significantly deeper intestinal crypts and a thinner lamina propria layer. Pigs fed hLZ-milk, rhLF-milk and rhLF+hLZ had significantly reduced mean corpuscular volume (MCV) and red blood cells (RBCs) were significantly increased in pigs fed hLZ-milk and rhLF-milk and tended to be increased in rhLF+hLZ-fed pigs, indicating more mature RBCs. These results support previous research demonstrating that pigs fed milk containing rhLF or hLZ had decreased intestinal inflammation, and suggest that in some parameters the combination of lactoferrin and lysozyme have additive effects, in contrast to the synergistic effects reported when utilising in-vitro models.

Despite the proven ability of infrared thermography (IRT) technology for early detection of subclinical mastitis in dairy cows/sheep/goats, studies on its diagnostic feasibility in dairy camels are lacking. Sixty-five lactating camels in mid lactation, machine milked twice-daily and managed under intensive conditions were screened to evaluate the feasibility of utilising IRT compared with other routine indicators in detecting subclinical mastitis. Immediately before the morning milking, a portable infrared camera was used to obtain thermograms in duplicate for the front and rear left quarters to determine the udder surface temperature (UST). Thereafter, milk samples from quarters were collected, and processed for California mastitis test (CMT) score and somatic cell count (SCC). In the present study, CMT score was used to define subclinical mastitis and the feasibility of IRT to detect subclinical mastitis was compared with CMT and SCC. According to CMT score, subclinical mastitic udders had an average UST of 1·42 °C greater (P<0·0001) than healthy udders. The relationship between UST and CMT was found to be highly correlated (r=0·77; P<0·001), and UST was linearly increased as the CMT increased [UST (°C)=34·86+0·66×CMT score; R2=0·65; P<0·001]. Additionally, high correlations were obtained between UST and SCC score (r=0·75; P<0·001) and the relationship between UST and SCC was fitted best to a logarithmic equation [UST, °C=33·39+0·88×Log (SCC, ×103 cells/ml), R2=0·61, P<0·001]. The cut-off value, sensitivity, specificity and area under the receiver operating characteristics (ROC) curve for UST were 35·15 °C, 0·90, 0·98, and 0·95, respectively, when subclinical mastitis was defined according to CMT score, and were 35·70 °C, 0·89, 0·96 and 0·94, respectively, when categorised according to the obtained SCC threshold (SCC=432 000 cells/ml). In conclusion, IRT, as an indirect non-invasive screening method, was highly feasible for distinguishing subclinical mastitic udders in dairy camels, which is crucial to treat mastitis early and efficiently.

Each cheese producer is responsible by the legislation for the number of Listeria monocytogenes in cheese and is required to prove that numbers will not exceed 100 cfu/g throughout the shelf-life of the cheese. Even in the case of hard-cheese such as Cheddar cheese, the absence of growth of List. monocytogenes during ripening has to be demonstrated to comply with EU legislation. Studies dedicated to assessing List. monocytogenes growth throughout cheese shelf-life are generally based on artificially contaminated cheeses. Contrary to the majority of works, the current study focused on the growth of List. monocytogenes in naturally contaminated raw milk farmhouse Cheddar cheeses during a five-month ripening period. List. monocytogenes growth was assessed by direct count and its presence was detected by enrichment in two naturally contaminated cheese batches. In order to track routes of contamination, 199 processing environment samples from inside and outside the processing facility were taken, and their analysis for the presence of List. monocytogenes was performed on four occasions over a 9-month period. List. monocytogenes isolates were differentiated using PFGE and serotyping. List. monocytogenes never exceeded 20 cfu/g in the cheeses and could not be detected after five months of ripening. Eleven pulsotypes were identified. One pulsotype was found in the yard outside the processing facility, in a vat, on the processing area floor and in a cheese. This indicated that the outside environment constitutes a potential source of contamination of the processing environment and of the cheese. These results demonstrate that this farmhouse Cheddar cheese does not support List. monocytogenes growth and suggests that the efforts to reduce processing environment contamination are worthwhile.

The internal teat sealant OrbeSeal® (Zoetis, Berlin, Germany) is intended to prevent new intramammary infection (IMI) throughout the dry period. The aim of this field study was to determine new infection rates in udder quarters applied exclusively with the sealant at dry-off and untreated ones (control group). For that, the new infection rate of udder quarters treated with the sealant was evaluated in a split-udder design (front right and rear left udder quarters treated, the other ones untreated) in 128 cows from nine dairy farms in North Germany that were healthy in all four udder quarters (i.e. <100 000 cells/ml, bacteriologically negative). After calving, duplicate quarter milk samples were collected twice at DIM (days in milk) 5–12 and 7 d later for culture. The new infection rates for treated and untreated groups were 3·4 and 10·5%, respectively. The results showed that applying a standardised study design with very few confounders under field conditions, using internal teat sealants reduced the new infection rate on quarter level to a third of that of the control group. Thus, internal teat sealants represent a viable option for dry-off treatment of udder-healthy cows.

This study was aimed to optimise lactose, inulin and yeast extract concentration and also culture pH for maximising the growth of a probiotic bacterium, Bifidobacterium animalis subsp. lactis in apple juice and to assess the effects of these factors by using response surface methodology. A second-order central composite design was applied to evaluate the effects of these independent variables on growth of the microorganism. A polynomial regression model with cubic and quadratic terms was used for analysis of the experimental data. It was found that the effects involving inulin, yeast extract and pH on growth of the bacterium were significant, and the strongest effect was given by the yeast extract concentration. Estimated optimum conditions of the factors on the bacterial growth are as follows: lactose concentration=9·5 g/l; inulin concentration=38·5 mg/l; yeast extract concentration=9·6 g/l and initial pH=6·2.

Lactation physiology is a process that is only partly understood. Proteomics techniques have shown to be useful to help advance the knowledge on lactation physiology in human and rodent species but have not been used as major tools for dairy cows, except for mastitis. In this paper, advanced non-targeted proteomics techniques (Filter aided sample preparation and NanoLC-Orbitrap-MS/MS) were applied to study the milk fat globule membrane and milk serum fraction, resulting in the identification of 246 proteins. Of these, 23 transporters and enzymes were related to lipid synthesis and secretion in mammary gland and their functions are discussed in detail. The identification of these intracellular transporters and enzymes in milk provides a possibility of using milk itself to study lipid synthesis and secretion pathways. This full-scale scan of milk proteins by using non-targeted proteomic analysis helps to reveal the important proteins involved in lipid synthesis and secretion for further examination in targeted studies.

Colostrum consists of a number of biologically active proteins and peptides that influence physiological function and development of a neonate. The present study investigated the biological activity of peptides released from first day bovine colostrum through in vitro and in vivo enzymatic digestion. This was assessed for proliferative activity using a human intestinal epithelial cell line, T84. Digestion of the protein fraction of bovine colostrum in vitro was conducted with the enzymes pepsin, chymosin and trypsin. Pepsin and chymosin digests yielded protein fractions with proliferative activity similar to that observed with undigested colostrum and the positive control foetal calf serum (FCS). In contrast trypsin digestion significantly (P<0·05) decreased colostral proliferative activity when co-cultured with cells when compared with undigested colostrum. The proliferative activity of undigested colostrum protein and abomasal whey protein digesta significantly increased (P<0·05) epithelial cell proliferation in comparison to a synthetic peptide mix. Bovine colostrum protein digested in vivo was collected from different regions of the gastrointestinal tract (GIT) in newborn calves fed either once (n=3 calves) or three times at 12-h intervals (n=3 calves). Digesta collected from the distal duodenum, jejunum and colon of calves fed once, significantly (P<0·05) stimulated cell proliferation in comparison with comparable samples collected from calves fed multiple times. These peptide enriched fractions are likely to yield candidate peptides with potential application for gastrointestinal repair in mammalian species.

To evaluate the interaction between the levels of dietary concentrate and linseed oil (LO) on milk fatty acid (FA) profile, 24 Holstein cows were used in a randomised complete block design based on days in milk, with a 2×2 factorial arrangement of treatments. Within each block, cows were fed one of four experimental diets containing 30% concentrate (LC) or 70% concentrate (HC), without LO (NLO) or with LO supplemented at 3% of dietary dry matter. Milk FA profiles were analysed with a special emphasis on the intermediates of the predominant trans-11, and a putative trans-13 pathways of ruminal biohydrogenation of cis-9, cis-12, cis-15 18:3. Feeding LO increased the concentrations of cis-9, cis-12, cis-15 18:3 and trans-11, cis-15 18:2 in milk fat, and these increases were of a higher magnitude when LO was added in HC as compared with LC diet (interaction of LO by concentrate). A treatment interaction was also observed for the level of trans-11 18:1 which was higher when feeding LO, but for which the increase was more pronounced with the LC as compared with HC diet. The concentrations of cis-15 18:1 and cis-9, trans-11, cis-15 18:3 were higher in cows fed LO, but feeding HC diets decreased milk fat content of cis-15 18:1 and a tendency for a decrease in cis-9, trans-11, cis-15 18:3 was apparent. Feeding LO increased milk fat content of trans-13 18:1 and cis-9, trans-13 18:2, while the concentrations of these two isomers were not affected by the level of dietary concentrates. The isomer cis-9, trans-13, cis-15 18:3 has not been detected in any of the milk samples. In conclusion, interactions were observed between LO and dietary concentrates on the proportions of some intermediates of the trans-11 biohydrogenation pathway. The presence of trans-13 18:1 and cis-9, trans-13 18:2 supports the existence of a trans-13 pathway, but an 18:3 intermediate with a trans-13 double bond has not been identified.

Lactones are important contributors to the flavour and aroma of milk and dairy products. This study was conducted to evaluate the effects of dietary linseed oil (LO) and forage to concentrate ratio on milk lactone profile. Twenty four Holstein cows were used during a 4-week feeding trial in a randomised complete block design. Cows were fed diets containing 30% (LC) or 70% (HC) concentrate, and 0% (NLO) or 3% LO in a 2×2 factorial arrangement of treatments. Milk lactone profile was evaluated using the solid phase microextraction technique. The highest levels of δ-lactones (δ-6:0, δ-8:0, δ-10:0, and δ-12:0) were found with the LC/NLO diet. These concentrations were then decreased when cows received either a high level of concentrate or supplemental LO, but these effects were not additive (interaction of LO by concentrate, P<0·01). An interaction of LO by concentrate (P<0·01) was also noted on milk γ-12:0 for which the highest concentration was observed when supplementing LO in HC diet, while no effect was apparent when LO was added in LC diet. Moreover, feeding HC increased the level of γ-12:1 in milk as compared with LC, while LO had no effect on this γ-lactone. Finally, γ-12:2 was not detected in any of the milk samples studied. Organoleptic properties of milk were evaluated in a triangle test showing that a significant number of assessors perceived a difference between milk from cows fed LC/NLO as compared with milk from cows fed HC/LO. The sensory evaluation was completed by a ranking test where the intensities of fresh lactic, foreign and global flavours were not different between treatments. In conclusion, feeding LO in HC diet modified milk lactone profile with a shift toward more γ- and less δ-lactones as compared with LC diet not supplemented with LO. A difference was perceived in a triangle test between milk from these two treatments, but the sensory attributes responsible for this difference have not been identified in the current trial.

Alpha-lactalbumin (α-la) is one of the major proteins in whey. When partially hydrolysed with Bacillus licheniformis protease, it produces nanotubular structures in the presence of calcium ions by a self-assembly process. This study presents investigation of α-la protein structure during hydrolysis and nanotube formation using optical spectroscopy. Before spectroscopic measurements, nanotubes were examined with microscopy. The observed α-la nanotubes (α-LaNTs) were in the form of regular hollow strands with a diameter of about 20 nm and the average length of 1 μm. Amide and backbone vibration bands of the Raman spectra displayed remarkable conformational changes in α and β domains in the protein structure during nanotube growth. This was confirmed by the Fourier-transform infrared (FTIR) spectroscopy data. Also, FTIR analysis revealed certain bands at calcium (Ca++) binding sites of COO− groups in hydrolysed protein. These sites might be critical in nanotube elongation.

The goal of the present study was to examine the suitability of a short pre-stimulation (P) for 15 s followed by a latency period (L) of 30 s before cluster attachment for machine milking. In addition we tested the effect of a periodic reduction of the vacuum under the teat (VR) during the massage phase from 43 kPa to 12–15 kPa on milking characteristics and teat tissue condition. The study was carried out in 9 cows in a cross-over design. Animals were milked twice daily, and each of the 4 treatment combinations was used for six subsequent milkings (P+L vs. continuous P, and standard pulsation vs. VR, respectively). Milk flow was recorded during all experimental milkings. Longitudinal ultrasound cross sections of the teat were performed by B-mode ultrasound after the last milking of each treatment at 0, 5, and 15 min after the end of milking, respectively. None of the evaluated milking characteristics (total milk yield, main milking time, peak flow rate, average milk flow) differed between treatments. Teat measures as obtained by ultrasound cross sections showed no significant difference if individual treatments were compared at the three time points individually. However, teat wall thickness (TWT) tended to be smaller in VR vs. non-VR treatments at 5 min after milking (P=0·05). In conclusion, teat preparation consisting of a short stimulation followed by a latency period represents a similarly efficient pre-stimulation as a continuous pre-stimulation. VR seems to reduce the load on the teat tissue during milking and thus reduces the development of oedema and hence a less pronounced increase of TWT while milking characteristics are similar with or without VR.

The aim of the study was to investigate how the use of fresh cheese brines compared with used brines and various combinations of pH and NaCl concentrations affected the survival of Listeria monocytogenes. Cheese brines from five Norwegian small scale cheese producers were analysed and showed great variations in pH (4·54–6·01) and NaCl concentrations (14·1–26·9 %). The survival of five strains of List. monocytogenes (two clinical isolates, two food isolates and one animal isolate) in four different cheese brines (three used and one fresh) was investigated. Results showed significant differences in survival both depending on the strains and the brines. Strains of human outbreak listeriosis cases showed greater ability to survive in the brines compared with food isolates and a List. monocytogenes reference strain (1–2 log10 difference after 200 d). All strains showed highest survival in the freshly prepared brine compared with the used brines. Molecular typing by multiple locus variable number tandem repeats analysis (MLVA) showed that there were no detectable alterations in the examined variable number tandem repeats of the genome in five strains after 200 d storage in any of the salt brines. Combined effects of pH (4·5, 5·25 and 6·0) and NaCl (15, 20 and 25 %) in fresh, filter sterilised brines on the survival of List. monocytogenes were examined and results showed that pathogen populations decreased over time in all brines. Death rates at any given NaCl concentration were highest at low pH (4·5) and death rates at any given pH were highest at low NaCl concentrations (15 %). In conclusion, the use of used brines reduced the survival of List. monocytogenes and a combination of low pH (4·5) and low salt concentrations (15 %) decreased the risk of List. monocytogenes survival compared with higher pH (5·25 or 6·0) and higher NaCl concentrations (20 or 25 %).

The main goal of this experiment was to study the effect of milk fat depression, induced by supplementing diet with plant oils, on the bovine fat metabolism, with special interest in cholesterol levels. For this purpose 39 cows were divided in three groups and fed different rations: a control group (C) without any oil supplementation and two groups with soybean oil (SO) or rapeseed oil (RO) added to the partial mixed ration (PMR). A decrease in milk fat percentage was observed in both oil feedings with a higher decrease of −1·14 % with SO than RO with −0·98 % compared with the physiological (−0·15 %) decline in the C group. There was no significant change in protein and lactose yield. The daily milk cholesterol yield was lower in both oil rations than in control ration, while the blood cholesterol level showed an opposite variation. The milk fatty acid pattern showed a highly significant decrease of over 10 % in the amount of saturated fatty acids (SFA) in both oil feedings and a highly significant increase in mono (MUFA) and poly (PUFA) unsaturated fatty acids, conjugated linoleic acids (CLA) included. The results of this experiment suggest that the feeding of oil supplements has a high impact on milk fat composition and its significance for human health, by decreasing fats with a potentially negative effect (SFA and cholesterol) while simultaneously increasing others with positive (MUFA, PUFA, CLA).