Lactation represents an important element of the life history strategies of all mammals, whether monotreme, marsupial, or eutherian. Milk originated as a glandular skin secretion in synapsids (the lineage ancestral to mammals), perhaps as early as the Pennsylvanian period, that is, approximately 310 million years ago (mya). Early synapsids laid eggs with parchment-like shells intolerant of desiccation and apparently dependent on glandular skin secretions for moisture. Mammary glands probably evolved from apocrine-like glands that combined multiple modes of secretion and developed in association with hair follicles. Comparative analyses of the evolutionary origin of milk constituents support a scenario in which these secretions evolved into a nutrient-rich milk long before mammals arose. A variety of antimicrobial and secretory constituents were co-opted into novel roles related to nutrition of the young. Secretory calcium-binding phosphoproteins may originally have had a role in calcium delivery to eggs; however, by evolving into large, complex casein micelles, they took on an important role in transport of amino acids, calcium and phosphorus. Several proteins involved in immunity, including an ancestral butyrophilin and xanthine oxidoreductase, were incorporated into a novel membrane-bound lipid droplet (the milk fat globule) that became a primary mode of energy transfer. An ancestral c-lysozyme lost its lytic functions in favor of a role as α-lactalbumin, which modifies a galactosyltransferase to recognize glucose as an acceptor, leading to the synthesis of novel milk sugars, of which free oligosaccharides may have predated free lactose. An ancestral lipocalin and an ancestral whey acidic protein four-disulphide core protein apparently lost their original transport and antimicrobial functions when they became the whey proteins β-lactoglobulin and whey acidic protein, which with α-lactalbumin provide limiting sulfur amino acids to the young. By the late Triassic period (ca 210 mya), mammaliaforms (mammalian ancestors) were endothermic (requiring fluid to replace incubatory water losses of eggs), very small in size (making large eggs impossible), and had rapid growth and limited tooth replacement (indicating delayed onset of feeding and reliance on milk). Thus, milk had already supplanted egg yolk as the primary nutrient source, and by the Jurassic period (ca 170 mya) vitellogenin genes were being lost. All primary milk constituents evolved before the appearance of mammals, and some constituents may have origins that predate the split of the synapsids from sauropsids (the lineage leading to ‘reptiles’ and birds). Thus, the modern dairy industry is built upon a very old foundation, the cornerstones of which were laid even before dinosaurs ruled the earth in the Jurassic and Cretaceous periods.

Mammalian milk or colostrum contains up to 10% of carbohydrate, of which free lactose usually constitutes more than 80%. Lactose is synthesized within lactating mammary glands from uridine diphosphate galactose (UDP-Gal) and glucose by a transgalactosylation catalysed by a complex of β4-galactosyltransferase and α-lactalbumin (α-LA). α-LA is believed to have evolved from C-type lysozyme. Mammalian milk or colostrum usually contains a variety of oligosaccharides in addition to free lactose. Each oligosaccharide has a lactose unit at its reducing end; this unit acts as a precursor that is essential for its biosynthesis. It is generally believed that milk oligosaccharides act as prebiotics and also as receptor analogues that act as anti-infection factors. We propose the following hypothesis. The proto-lacteal secretions of the primitive mammary glands of the common ancestor of mammals contained fat and protein including lysozyme, but no lactose or oligosaccharides because of the absence of α-LA. When α-LA first appeared as a result of its evolution from lysozyme, its content within the lactating mammary glands was low and lactose was therefore synthesized at a slow rate. Because of the presence of glycosyltransferases, almost all of the nascent lactose was utilized for the biosynthesis of oligosaccharides. The predominant saccharides in the proto-lacteal secretions or primitive milk produced by this common ancestor were therefore oligosaccharides rather than free lactose. Subsequent to this initial period, the oligosaccharides began to serve as anti-infection factors. They were then recruited as a significant energy source for the neonate, which was achieved by an increase in the synthesis of α-LA. This produced a concomitant increase in the concentration of lactose in the milk, and lactose therefore became an important energy source for most eutherians, whereas oligosaccharides continued to serve mainly as anti-microbial agents. Lactose, in addition, began to act as an osmoregulatory molecule, controlling the milk volume. Studies on the chemical structures of the milk oligosaccharides of a variety of mammalian species suggest that human milk or colostrum is unique in that oligosaccharides containing lacto-N-biose I (LNB) (Gal(β1 → 3)GlcNAc, type I) predominate over those containing N-acetyllactosamine (Gal(β1 → 4)GlcNAc, type II), whereas in other species only type II oligosaccharides are found or else they predominate over type I oligosaccharides. It can be hypothesized that this feature may have a selective advantage in that it may promote the growth of beneficial colonic bacteria, Bifidobacteria, in the human infant colon.

A potential role for epigenetic mechanisms in the regulation of mammary function in the dairy cow is emerging. Epigenetics is the study of heritable changes in genome function that occur because of chemical changes rather than DNA sequence changes. DNA methylation is an epigenetic event that results in the silencing of gene expression and may be passed on to the next generation. However, recent studies investigating different physiological states and changes in milk protein gene expression suggest that DNA methylation may also play an acute, regulatory, role in gene transcription. This overview will highlight the role of DNA methylation in the silencing of milk protein gene expression during mastitis and mammary involution. Moreover, environmental factors such as nutrition may induce epigenetic modifications of gene expression. The current research investigating the possibility of in utero, hence cross-generational, epigenetic modifications in dairy cows will also be discussed. Understanding how the mammary gland responds to environmental cues provides a potential to enhance milk production not only of the dairy cow but also of her daughter.

Mammary stem cells (MaSC) provide for net growth, renewal and turnover of mammary epithelial cells, and are therefore potential targets for strategies to increase production efficiency. Appropriate regulation of MaSC can potentially benefit milk yield, persistency, dry period management and tissue repair. Accordingly, we and others have attempted to characterize and alter the function of bovine MaSC. In this review, we provide an overview of current knowledge of MaSC gained from studies using mouse and human model systems and present research on bovine MaSC within that context. Recent data indicate that MaSC retain labeled DNA for extended periods because of their selective segregation of template DNA strands during mitosis. Relying on this long-term retention of bromodeoxyuridine-labeled DNA, we identified putative bovine MaSC. These label-retaining epithelial cells (LREC) are in low abundance within mammary epithelium (<1%). They are predominantly estrogen receptor (ER)-negative and localized in a basal or suprabasal layer of the epithelium throughout the gland. Thus, the response of MaSC to estrogen, the major mitogen in mammary gland, is likely mediated by paracrine factors released by cells that are ER-positive. This is consistent with considerable evidence for cross-talk within and between epithelial cells and surrounding stromal cells. Excision of classes of cells by laser microdissection and subsequent microarray analysis will hopefully provide markers for MaSC and insights into their regulation. Preliminary analyses of gene expression in laser-microdissected LREC and non-LREC are consistent with the concept that LREC represent populations of stem cells and progenitor cells that differ with regard to their properties and location within the epithelial layer. We have attempted to modulate the MaSC number by infusing a solution of xanthosine through the teat canal and into the ductal network of the mammary glands of prepubertal heifers. This treatment increased the number of putative stem cells, as evidenced by an increase in the percentage of LREC and increased telomerase activity within the tissue. The exciting possibility that stem cell expansion can influence milk production is currently under investigation.

Environmental variables such as photoperiod, heat, stress, nutrition and other external factors have profound effects on quality and quantity of a dairy cow's milk. The way in which the environment interacts with genotype to impact milk production is unknown; however, evidence from our laboratory suggests that circadian clocks play a role. Daily and seasonal endocrine rhythms are coordinated in mammals by the master circadian clock in the hypothalamus. Peripheral clocks are distributed in every organ and coordinated by signals from the master clock. We and others have shown that there is a circadian clock in the mammary gland. Approximately 7% of the genes expressed during lactation had circadian patterns including core clock and metabolic genes. Amplitude changes occurred in the core mammary clock genes during the transition from pregnancy to lactation and were coordinated with changes in molecular clocks among multiple tissues. In vitro studies using a bovine mammary cell line showed that external stimulation synchronized mammary clocks, and expression of the core clock gene, BMAL1, was induced by lactogens. Female clock/clock mutant mice, which have disrupted circadian rhythms, have impaired mammary development and their offspring failed to thrive suggesting that the dam's milk production was not adequate enough to nourish their young. We envision that, in mammals, during the transition from pregnancy to lactation the master clock is modified by environmental and physiological cues that it receives, including photoperiod length. In turn, the master clock coordinates changes in endocrine milieu that signals peripheral tissues. In dairy cows, it is clear that changes in photoperiod during the dry period and/or during lactation influences milk production. We believe that the photoperiod effect on milk production is mediated, in part by the ‘setting’ of the master clock with light, which modifies peripheral circadian clocks including the mammary core clock and subsequently impacts milk yield and may impact milk composition.

The dry period is required to facilitate cell turnover in the bovine mammary gland in order to optimize milk yield in the next lactation. Traditionally, an 8-week dry period has been a standard management practice for dairy cows based on retrospective analyses of milk yields following various dry period lengths. However, as milk production per cow has increased, transitioning cows from the nonlactating state to peak milk yield has grown more problematic. This has prompted new studies on dry period requirements for dairy cows. These studies indicate a clear parity effect on dry period requirement. First parity animals require a 60-day dry period, whereas lactations following later parities demonstrate no negative impact with 30-day dry period or even eliminating the dry period when somatotropin (ST) is also used to maintain milk yields. Shortened dry periods in first parity animals were associated with reduced mammary cell turnover during the dry period and early lactation and increased numbers of senescent cells and reduced functionality of lactating alveolar mammary cells postpartum. Use of ST and increased milking frequency postpartum reduced the impact of shortened dry periods. The majority of new intramammary infections occur during the dry period and persist into the following lactation. There is therefore the possibility of altering mastitis incidence by modifying or eliminating the dry period in older parity animals. As the composition of mammary secretions including immunoglobulins may be reduced when the dry period is reduced or eliminated, there is the possibility that the immune status of cows during the peripartum period is influenced by the length of the dry period.

Milk is a source of bioactive molecules with wide-ranging functions. Among these, the immune properties have been the best characterised. In recent years, it has become apparent that besides the immunoglobulins, milk also contains a range of minor immune-related proteins that collectively form a significant first line of defence against pathogens, acting both within the mammary gland itself as well as in the digestive tract of the suckling neonate. We have used proteomics technologies to characterise the repertoire of host-defence-related milk proteins in detail, revealing more than 100 distinct gene products in milk, of which at least 15 are known host-defence-related proteins. Those having intrinsic antimicrobial activity likely function as effector proteins of the local mucosal immune defence (e.g. defensins, cathelicidins and the calgranulins). Here, we focus on the activities and biological roles of the cathelicidins and mammary serum amyloid A. The function of the immune-related milk proteins that do not have intrinsic antimicrobial activity is also discussed, notably lipopolysaccharide-binding protein, RNase4, RNase5/angiogenin and cartilage-glycoprotein 39 kDa. Evidence is shown that at least some of these facilitate recognition of microbes, resulting in the activation of innate immune signalling pathways in cells associated with the mammary and/or gut mucosal surface. Finally, the contribution of the bacteria in milk to its functionality is discussed. These investigations are elucidating how an effective first line of defence is achieved in the bovine mammary gland and how milk contributes to optimal digestive function in the suckling calf. This study will contribute to a better understanding of the health benefits of milk, as well as to the development of high-value ingredients from milk.

The life science industries have a strong interest in screening for novel bioactives in complex mixtures like milk and dairy products. Food bioactives are not only important for public health in general, but also have potential therapeutic applications for the treatment of a number of diseases. To identify these novel bioactives, establishment of robust screening assays is essential. The use of in vitro cell-based models for screening and testing have the advantage that several concentrations of mixtures or specific compounds can be assayed at the same time in cells from specific tissues. Primary cell cultures from target organs or established cell lines can be used to identify the most sensitive cells. In addition, a large number of transfected cell lines with very specific sensitivities have been developed. Different endpoints inherent to basal or more sophisticated cellular functions can be investigated, such as cell viability, apoptosis, migration, intracellular signalling, regulation of gene expression, morphology and metabolic alterations. The gastrointestinal tract is an obvious target for bioactive molecules delivered through milk and dairy products, because it lies at the interface between dietary components in the lumen and the internal processes of the host. Identification of bioactive factors that affects proliferation or migration of epithelial cells may have potential applications in promoting gastrointestinal health in both humans and animals. The mammary gland is another target organ of considerable interest since it has been estimated that up to 50% of all newly diagnosed breast cancers may be related to dietary factors. A large number of gastrointestinal and mammary epithelial cell lines are commercially available, but in order to study some cellular functions, primary cultures of freshly isolated cells are often preferred, as established cell lines do not always express specialised properties in culture.

In this study, random regression models were used to estimate covariance functions between feed intake and BW in boars from the two breeds: the Norwegian Landrace and the Norwegian Duroc. In total, 1476 animals of the Norwegian Landrace breed and 1300 animals of the Norwegian Duroc breed had registrations on daily feed intake and growth from 54 to 180 days of age. Random regressions on the Legendre polynomials of age were used to describe genetic and permanent environmental curves in BW (up to the second order) and feed intake (up to the first order) for both the breeds. Heritabilities on BW increased over time for the Landrace (0.18 to 0.24), but were approximately constant for the Duroc (0.33 to 0.35). Average heritabilities for feed intake were approximately the same in both the breeds (0.09 to 0.11), and the estimates decreased over time, most pronounced in Duroc. On the basis of the current data, daily feed intake was seemingly controlled by the same genetic factors throughout the test period for Duroc; however, for Landrace, genetic correlations between test days decreased with increasing distance in time. For BW, the genetic correlations between test days were in general high, and did not go below 0.8 for any of the two breeds in this study. For both feed intake and BW, permanent environmental correlations between start and end of the test were reduced with increasing difference in days, most pronounced in Duroc. This study indicates that weight of the animal at the end of the test was more closely genetically correlated to feed intake of earlier periods compared with later periods of growth for both the breeds. This may be explained by the fact that BW is the cumulative growth of an individual, which is likely to be heavily affected by the feed intake during the most intense growth period.

The objective of this study was to carry out a sensitivity analysis on the impact of various production strategies and performance levels on the relative economic values (REVs) of traits in dairy sheep. A bio-economic model implemented in the program package ECOWEIGHT was used to simulate the profit function for a semi-extensive production system with the Slovak multi-purpose breed Improved Valachian and to calculate the REV of 14 production and functional traits. The following production strategies were analysed: differing proportions of milk processed to cheese, customary weaning and early weaning of lambs with immediate sale or sale after artificial rearing, seasonal lambing in winter and aseasonal lambing in autumn. Results of the sensitivity analysis are presented in detail for the four economically most important traits: 150 days milk yield, conception rate of ewes, litter size and ewe productive lifetime. Impacts of the differences in the mean value of each of these four traits on REVs of all other traits were also examined. Simulated changes in the production circumstances had a higher impact on the REV for milk yield than on REVs of the other traits investigated. The proportion of milk processed to cheese, weaning management strategy for lambs and level of milk yield were the main factors influencing the REV of milk yield. The REVs for conception rate of ewes were highly sensitive to the current mean level of the trait. The REV of ewe productive lifetime was most sensitive to variation in ewe conception rate, and the REV of litter size was most affected by weaning strategy for lambs. On the basis of the results of sensitivity analyses, it is recommended that economic values of traits for the overall breeding objective for dairy sheep be calculated as the weighted average of the economic values obtained for the most common production strategies of Slovak dairy sheep farms and that economic values be adjusted after substantial changes in performance levels of the traits.

To investigate the potential use of n-alkanes (alkanes), long-chain alcohols (alcohols) and long-chain fatty acids (acids) for estimating the diet composition of sheep, in a feeding trial. A total of 18 sheep were assigned randomly to three different diets (diet A, diet B and diet C) containing up to eight herbage species (Leymus chinensis, Leymus dasystachys, Elymus sibiricum, Chenopodium album, Puccinellia chinampoensis, Medicago sativa, Saussurea sinuata and Bromus inermis). Faecal recoveries of alkanes, alcohols and acids were determined, and diet compositions were estimated using different combinations of alkanes, alcohols and acids. The faecal concentrations of individual alkanes, alcohols and acids were corrected using the mean recovery of the dietary treatment that the respective animal belonged to (diet recovery), or the mean recovery across all dietary treatments (general recovery). In general, diets did not affect the faecal recovery values for alkanes, alcohols and acids, and no difference in accuracy was found between diet composition estimates based on dietary recovery and general recovery. The accuracy of diet composition estimates declined as the number of dietary components increased from four to eight herbage species (P < 0.001). Better (P < 0.05) estimates of diet composition were obtained with the combinations of two or three marker types instead of alkanes alone. Moreover, results showed that excluding minor diet components from the calculations decreased (P < 0.05) the accuracy of diet composition estimates, whereas including extra non-grazed herbage species did not reduce (P > 0.05) the quality of diet composition estimates. These results confirmed the usefulness of alkanes, alcohols and acids as markers for determining complex diet composition of sheep. However, a negative impact on the accuracy of diet composition estimates, caused by missing minor diet components from the calculation of diet composition, could happen when plant wax markers are used to estimate the diet composition of free-ranging animals.

On the basis of the isomer-specific effects of trans fatty acids (FA) on human health, and the detrimental effect of t10,c12-conjugated linoleic acid (CLA) on cows’ milk fat production, there is a need to identify factors that affect the shift from trans-11 to trans-10 pathway during ruminal biohydrogenation of FA. This experiment was conducted in vitro and aimed at separating the effects of the diet of the donor cows from those of the fermentative substrate, which is necessary to prevent this shift. A total of four dry Holstein dairy cows were used in a 4 × 4 Latin square design. They received 12 kg of dry matter per day of four diets based on maize silage during four successive periods: the control diet (22% starch, <3% fat); the high-starch diet, supplemented with wheat plus barley (35% starch, <3% crude fat); the sunflower oil diet, supplemented with 5% of sunflower oil (20% starch, 7.6% crude fat); and the high-starch plus oil diet (33% starch, 7.3% crude fat). Ruminal fluid of each donor cow was incubated for 5 h with four substrates having similar chemical composition to the diets, replacing sunflower oil by pure linoleic acid (LA). The efficiency of isomerisation of LA to CLA was the highest when rumen fluids from cows receiving dietary oil were incubated with added LA. The shift from trans-11 to trans-10 isomers was induced in vitro by high-starch diets and the addition of LA. Oil supplementation to the diet of the donor cows increased this shift. Conversely, the trans-10 isomer balance was always low when no LA was added to incubation cultures. These results showed that a large accumulation of trans-10 FA was only observed with an adapted microflora, as well as an addition of non-esterified LA to the incubation substrate.

Previous studies have indicated that 15N enrichment of solid-associated bacteria (SAB) may be predicted from the same value in liquid-associated bacteria (LAB). The aims of this study were to confirm this and to measure the error in the nutrient supply from SAB, when LAB are used as the reference sample. For this purpose, the chemical and amino acid (AA) compositions of both the bacterial populations were studied in four experiments carried out on different groups of three rumen cannulated wethers. Diets (one in Experiments 1 and 4 and three in Experiments 2 and 3) had forage-to-concentrate ratios (dry matter (DM) basis) between 2 : 1 and 40 : 60, and were consumed at intake levels between 40 and 75 g DM/kg (BW)0.75. The bacteria samples were isolated after continuous infusion of (15NH4)2SO4 (40, 18, 30 and 25 mg 15N/day, in Experiments 1 to 4, respectively) for at least 14 days. In all experiments, SAB had consistently higher concentrations of organic matter (826 v. 716 g/kg DM, as average) and total lipids (192 v. 95 g/kg DM, as average) than LAB. Similar CP concentrations of both populations were observed, except a higher concentration in SAB than in LAB in Experiment 3. A consistent (in Experiment 4 only as tendency) higher AA-N/total N ratio (on average 17.5%) was observed in SAB than in LAB. The 15N enrichment in SAB was systematically lower than in LAB. On the basis of the results of all studies a close relationship was found between the 15N enrichment in SAB and LAB, which was shown irrespective of experiments. This relationship was established from Experiments 1 and 2 and the above cited previous results (n = 20; P < 0.001; R2 = 0.996), and then confirmed from the results of Experiments 3 and 4. These relationships between SAB and LAB demonstrate that CP supply from SAB is underevaluated by, on average, 21.2% when LAB are used as the reference. This underevaluation was higher for true protein and even higher for the lipid supply (32.5% and 59.6%, respectively, as an average of the four experiments). Large differences in AA profile were observed between SAB and LAB. The prediction equation obtained using 15N as the marker may be used to correct the errors associated with the traditional use of LAB as the reference sample, and therefore to obtain more accurate estimates of the microbial nutrient supply to the ruminants.

The overall reproductive performance has decreased over the last decades, involving changes in cyclicity, oestrous behaviour and fertility. High milk yield (MY), low body condition score (BCS) and large body condition (BC) loss have been identified as risk factors. However, these effects are often confounded, as high MY and body lipid reserve mobilization are correlated. The aim of this study was to evaluate the respective effects of MY and BC on post-partum ovarian cyclicity, oestrus and fertility of Holstein cows. This study provides novel information, as MY and BC change were uncoupled in the overall dataset that included 98 lactations and milk progesterone profiles. Cows were assigned to two feeding-level groups: high feed, which achieved high MY and moderate BC loss throughout lactation (8410 kg, −1.17 unit from calving to nadir BCS), and low feed, which limited MY and triggered a large BC loss (5719 kg, −1.54 unit). MY and BC had different effects at different stages of the reproductive process. Cyclicity as well as non-fertilization and early embryo mortality were mainly driven by body lipid reserves, whereas oestrous behaviour and late embryo mortality were related to MY. The results point to possible uncoupling between cyclicity, oestrus and early and late embryo survival allowing compensation along the reproductive process and leading to similar final reproductive performance. In compact calving systems, which require high pregnancy rates within a short period, higher MY strategies appear unsuitable even where BCS is maintained, owing to depressed oestrous behaviour and probably increased late embryo mortality, which delays rebreeding. Similarly, strategies that compromise cyclicity and fertility by excessively low BCS are unsuitable.

Primary bovine mammary epithelial cells (BMEC) were cultured in media containing varying concentrations of glucose, to determine the effects of glucose availability on glucose transport and its mechanism in bovine mammary gland. The BMEC incubated with 10 and 20 mM glucose had twofold greater glucose uptake than that with 2.5 mM glucose (P < 0.05). Increased glucose availability enhanced the cell proliferation (P < 0.05). As the glucose uptake is mediated by facilitative glucose transporters (GLUTs), the expression of GLUT mRNA was investigated. Compared with the control (2.5 mM), 5 and 10 mM glucose did not influence the abundance of GLUT1 mRNA (P < 0.05), whereas 20 mM glucose decreased the GLUT1 mRNA expression in the BMEC (P < 0.05). The expression of GLUT8 mRNA was not affected by any concentration of glucose (P > 0.05). As GLUTs are coupled with hexokinases (HKs) in regulating glucose uptake, the expression of HKs and their activities were also studied. The HK activity was greater in 5, 10 and 20 mM glucose than that in 2.5 mM glucose (P < 0.05). The expression of HK2 mRNA rather than HK1 mRNA was detected in the BMEC; however, the abundance of HK2 mRNA was not elevated by any concentrations of glucose compared with control (P > 0.05). Furthermore, addition of 3-bromopyruvate (30, 50 or 70 μM), an inhibitor of HK2, resulted in the decrease of glucose uptake and cell proliferation at both 2.5 and 10 mM glucose (P < 0.05). Therefore, the glucose concentrations may affect glucose uptake partly by altering the activity of HKs, and HK2 may play an important role in the regulation of glucose uptake in the BMEC.

Farrowing is an intrinsically risky process for both the sow and the piglets that can cause welfare and economic problems. The effects of the non-steroidal anti-inflammatory drug meloxicam on post-farrowing behaviour of sows, and the performance of piglets were investigated. A total of 48 sows were randomly allocated at the day of farrowing (day 0) into two homogeneous groups regarding parity, and treated with either meloxicam or saline solution as placebo. For each sow, number of position changes, total time lying and standing or sitting, feed intake and rectal temperature (RT) were recorded during 3 days after farrowing. Piglets were individually weighed at farrowing and at weaning. The number of position changes did not show significant differences between treatments (P = 0.79). Sows spent significantly less time lying during day +3 after farrowing in the meloxicam group than in the placebo group (P = 0.04). Feed intake and RT showed a parity effect (P < 0.001 in both cases); however, no treatment effect was observed (P = 0.67 and P = 0.47, respectively). Pre-weaning mortality rate in piglets was not affected by treatment. In litters from multiparous sows, piglets of low birth weight (defined as percentile 15: BW <1180 g) had an average daily gain significantly higher in the meloxicam group than in the placebo group (196.6 ± 7.2 v. 166.6 ± 9.1 g/day; P = 0.03). Although the administration of meloxicam 90 min after farrowing showed a positive effect on the total time lying of the sows, additional investigations are required to better qualify relevant indicators of pain following farrowing in sows and to specify the analgesic effects of meloxicam on piglet performance.

Bruises in cattle develop after the application of force, and they provide evidence for sub-optimal animal welfare. The aim of this study was to describe the gross characteristics of bruises in cows arriving at the slaughterhouse directly from farms or through the livestock market. The number of bruises and their distribution on the carcass as well as their severity, shape, size and colour were assessed post mortem in a slaughterhouse in Chile. A total of 258 cow carcasses were evaluated, and a total of 846 bruises were found on 243 of the carcasses. Cows that had passed through a livestock market (M-carcasses) had in total 563 bruises (mean 3.8 bruises/carcass, s.d. 2.0), whereas cows transported directly from farms (F-carcasses) had in total 283 bruises (mean 2.5 bruises/carcass, s.d. 1.8). The backs of F-carcasses had twice as many bruises as M-carcasses (32.9% and 16.2%, respectively), whereas bruises in the rib area were more frequently observed in M-carcasses (13.1%) than in F-carcasses (8.1%). Superficial bruises (grade 1) were the most frequently observed (66.2% of all bruises). Regarding the size of the bruises, 64 (7.6%) were classified as large, 271 (32.0%) as medium and 511 (60.4%) as small. Irregularly shaped bruises were the most frequent (91.1%, n = 771), followed by linear (3.8%, n = 32), circular (3.1%, n = 26) and tramline-shaped bruises (1.9%, n = 16). The latter were noticed only in M-carcasses, which may indicate that these animals were beaten more frequently with sticks or other rod-shaped objects. Fresh, bright red-coloured bruises were found more frequently on all the animals (69.5% from farms and 70.5% from market) compared with bluish (29.7% and 29.3%, respectively) and yellow bruises (0.4% and 0.2%, respectively). The method of selling was significantly associated with the number of bruises on the carcass (P < 0.001) and the anatomical site (P < 0.05), but not with the severity, shape and colour of the bruises. Increased fat coverage reduced the severity of bruises (P < 0.001). This study shows that, in Chile, market animals have more bruises than those sourced directly from farms, and their distribution is different. More information about the causes of infliction may help reduce bruises and it may also improve their welfare. Further studies are required to elucidate whether the causes of the high bruising in the case of animals passing through markets are related only to extra handling (repeated loading, unloading, transportation, eventual mixing) or to the way of handling by personnel and inadequate design.

Since they were first described in 1993, it was found that recombinant variable fragments (rVHHs) of heavy-chain antibodies (HCAbs) from Camelidae have unusual biophysical properties, as well as a special ability to interact with epitopes that are cryptic for conventional Abs. It has been assumed that in vivo raised polyclonal HCAbs (pHCAbs) should behave in a similar manner than rVHHs; however, this assumption has not been tested sufficiently. Furthermore, our own preliminary work on a single serum sample from a llama immunized with a β-lactamase, has suggested that pHCAbs have no special ability to down-modulate catalytic activity. In this work, we further explored the interaction of pHCAbs from four llamas raised against two microbial enzymes and analyzed it within a short and a long immunization plan. The relative contribution of pHCAbs to serum titer was found to be low compared with that of the most abundant conventional subisotype (IgG1), during the whole immunization schedule. Furthermore, pHCAbs not only failed to inhibit the enzymes, but also activated one of them. Altogether, these results suggest that raising high titer inhibitory HCAbs is not a straightforward strategy – neither as a biotechnological strategy nor in the biological context of an immune response against infection – as raising inhibitory rVHHs.

The release rate (RR) of sulphur hexafluoride (SF6) gas from permeation tube in the rumen appears to be positively related with methane (CH4) emissions calculated using the SF6 tracer technique. Gas samples of breath and ruminal headspace were collected simultaneously in order to evaluate the hypothesis that transactions of SF6 in the rumen are the source for this relationship. Six non-lactating dairy cows fitted with rumen cannulae were subdivided into two groups and randomly assigned to a two-period crossover design to permeation tubes with low RR (LRR = 1.577 mg/day) or two-times higher RR (HRR = 3.147 mg/day) RR. The cows were fed limited amounts of maize silage (80% ad libitum) split into two meals (40% at 0800 h and 60% at 1600 h). Each period consisted of 3-day gas sampling. Immediately before the morning feed and then each hour over 8 h, ruminal gas samples (50 ml) were withdrawn through the cannula fitted with stoppers to prevent opening. Simultaneously, 8-h integrated breath gas samples were collected over the same period. Ratios of concentration of CH4/SF6, CO2/SF6 and CO2/CH4 and emission estimates of CH4 and CO2 were calculated for each sample source using the SF6 tracer technique principles. The LRR treatment yielded higher (P < 0.001) ruminal CH4/SF6 (by 1.79 times) and CO2/SF6 (by 1.90 times) ratios than the HRR treatment; however, these differences were lower than the 2.0 times difference expected from the RR between the LRR and HRR. Consequently, the LRR treatment was associated with lower (P < 0.01) ruminal emissions of CH4 over the 8-h collection period than with the HRR treatment (+11%), a difference also confirmed by the breath samples (+11%). RR treatments did not differ (P = 0.53) in ruminal or breath CO2 emissions; however, our results confirm that the SF6 tracer seems inappropriate for CO2 emissions estimation in ruminants. Irrespective of the RR treatment, breath samples yielded 8% to 9% higher CH4 emission estimates than the ruminal samples (P = 0.01). The relationship between rumen and breath sources for CH4 emissions was better for LRR than for HRR treatment, suggesting that tracer performance decreases with the highest RR of SF6 tested in our study (3.1 mg/day). A hypothesis is discussed with regard to the mechanism responsible for the relationship between RR and CH4 emission estimates. The use of permeation tubes with small range in RR is recommended in animal experiments to decrease variability in CH4 emission estimates using the SF6 tracer technique.