Estimates were obtained of the variability among herds of dairy cattle of the standard deviation (s.d.) and coefficient of variation (CV) of milk, fat and protein yield. For fat yield, for example, the mean s.d. within herds for heifers was about 30 kg, the s.d. among herds was 5·2 kg and the mean and s.d. of CV were 0·15 and 0·026, respectively. Correlations of both s.d. and CV were high between both heifers and cows in the same herd and year and between heifers or cows in the same herd in successive years, showing that variability was consistent over time and over age groups.

Ways of correcting for the heterogeneity of variance for use in cow index calculations are suggested; because CVs are heterogeneous, simple log transformation is not sufficient.

The pivotal roles of regulatory jurisdictions in the feed additive sector cannot be over-emphasized. In the European Union (EU), antioxidant substances are authorized as feed additives for prolonging the shelf life of feedstuffs based on their effect for preventing lipid peroxidation. However, the efficacy of antioxidants transcends their functional use as technological additives in animal feeds. Promising research results have revealed the in vivo efficacy of dietary antioxidants for combating oxidative stress in production animals. The in vivo effect of antioxidants is significant for enhancing animal health and welfare. Similarly, postmortem effect of dietary antioxidants has been demonstrated to improve the nutritional, organoleptic and shelf-life qualities of animal products. In practice, dietary antioxidants have been traditionally used by farmers for these benefits in livestock production. However, some antioxidants particularly when supplemented in excess could act as prooxidants and exert detrimental effects on animal well-being and product quality. Presently, there is no exclusive legislation in the EU to justify the authorization of antioxidant products for these in vivo and postmortem efficacy claims. To indicate these efficacy claims and appropriate dosage on product labels, it is important to broaden the authorization status of antioxidants through the appraisal of existing EU legislations on feed additives. Such regulatory review will have major impact on the legislative categorization of antioxidants and the efficacy assessment in the technical dossier application. The present review harnesses the scientific investigations of these efficacy claims in production animals and, proposes potential categorization and appraisal of in vivo methodologies for efficacy assessment of antioxidants. This review further elucidates the implication of such regulatory review on the practical application of antioxidants as feed additives in livestock production. Effecting these regulatory changes will stimulate the innovation of more potent antioxidant products and create potential new markets that will have profound economic impacts on the feed additive industry. Based on the in vivo efficacy claims, antioxidants may have to contend with the legislative controversy of either to be considered as veterinary drugs or feed additives. In this scenario, antioxidants are not intended to diagnose or cure diseases as ascribed to veterinary products. This twisted distinction can be logically debated with reference to the stipulated status of feed additives in Commission Regulation (EC) No 1831/2003. Nonetheless, it is imperative for relevant stakeholders in the feed additive industry to lobby for the review of existing EU legislations for authorization of antioxidants for these efficacy claims.

The networks generated by live animal movements are the principal vector for the propagation of infectious agents between farms, and their topology strongly affects how fast a disease may spread. The structural characteristics of networks may thus provide indicators of network vulnerability to the spread of infectious disease. This study applied social network analysis methods to describe the French swine trade network. Initial analysis involved calculating several parameters to characterize networks and then identifying high-risk subgroups of holdings for different time scales. Holding-specific centrality measurements (‘degree’, ‘betweenness’ and ‘ingoing infection chain’), which summarize the place and the role of holdings in the network, were compared according to the production type. In addition, network components and communities, areas where connectedness is particularly high and could influence the speed and the extent of a disease, were identified and analysed. Dealer holdings stood out because of their high centrality values suggesting that these holdings may control the flow of animals in part of the network. Herds with growing units had higher values for degree and betweenness centrality, representing central positions for both spreading and receiving disease, whereas herds with finishing units had higher values for in-degree and ingoing infection chain centrality values and appeared more vulnerable with many contacts through live animal movements and thus at potentially higher risk for introduction of contagious diseases. This reflects the dynamics of the swine trade with downward movements along the production chain. But, the significant heterogeneity of farms with several production units did not reveal any particular type of production for targeting disease surveillance or control. Besides, no giant strong connected component was observed, the network being rather organized according to communities of small or medium size (<20% of network size). Because of this fragmentation, the swine trade network appeared less structurally vulnerable than ruminant trade networks. This fragmentation is explained by the hierarchical structure, which thus limits the structural vulnerability of the global trade network. However, inside communities, the hierarchical structure of the swine production system would favour the spread of an infectious agent (especially if introduced in breeding herds).

Many studies with fin fish have demonstrated the potential to use alternative dietary protein sources to fish meal based on growth responses, although these trials mostly neglect to determine if such protein sources affect immune function. This study investigated the effect of fish meal replacement with dehulled lupin meal (LPN) or hydrolysed poultry feather meal (FTH). Atlantic salmon (Salmo salar L.) parr were supplied isonitrogenous and isoenergetic diets with 40% of the dietary protein provided by LPN or FTH, or 400 g/kg of the dietary protein provided equally by LPN and FTH (MIX). A diet mainly containing fish-meal protein acted as a control (CON). Growth, immune function, blood chemistry and disease resistance were assessed after 56 days. Significant differences (P < 0·05) in weight gain were detected between Atlantic salmon given the CON and FTH diets, whilst those salmon given LPN and MIX did not differ from any other. Productive protein values were significantly lower (P < 0·01) for salmon on FTH compared with those on CON and MIX. Immune function (as assessed by lysozyme, antiprotease, neutrophil oxygen radical production and plasma total immunoglobulin) and blood chemistry (as assessed by plasma total protein and glucose) were not significantly (P > 0·05) affected by any diet. Mortality rates of Atlantic salmon challenged with Vibrio anguillarum were not influenced by diet. These data suggest Atlantic salmon could be supplied diets with the fish meal component reduced to supply approximately 600 g/kg of the total protein, with the remaining 400 g/kg supplied by dehulled lupin meal or a combined dehulled lupin and hydrolysed poultry feather meal without any adverse effects on growth, immune function or blood chemistry.

This study was developed to assess the impact on performance, nutrient balance, serum parameters and feeding costs resulting from the switching of conventional to precision-feeding programs for growing–finishing pigs. A total of 70 pigs (30.4±2.2 kg BW) were used in a performance trial (84 days). The five treatments used in this experiment were a three-phase group-feeding program (control) obtained with fixed blending proportions of feeds A (high nutrient density) and B (low nutrient density); against four individual daily-phase feeding programs in which the blending proportions of feeds A and B were updated daily to meet 110%, 100%, 90% or 80% of the lysine requirements estimated using a mathematical model. Feed intake was recorded automatically by a computerized device in the feeders, and the pigs were weighed weekly during the project. Body composition traits were estimated by scanning with an ultrasound device and densitometer every 28 days. Nitrogen and phosphorus excretions were calculated by the difference between retention (obtained from densitometer measurements) and intake. Feeding costs were assessed using 2013 ingredient cost data. Feed intake, feed efficiency, back fat thickness, body fat mass and serum contents of total protein and phosphorus were similar among treatments. Feeding pigs in a daily-basis program providing 110%, 100% or 90% of the estimated individual lysine requirements also did not influence BW, body protein mass, weight gain and nitrogen retention in comparison with the animals in the group-feeding program. However, feeding pigs individually with diets tailored to match 100% of nutrient requirements made it possible to reduce (P<0.05) digestible lysine intake by 26%, estimated nitrogen excretion by 30% and feeding costs by US$7.60/pig (−10%) relative to group feeding. Precision feeding is an effective approach to make pig production more sustainable without compromising growth performance.

Regulation of foetal development in sheep depends on interactions between the intrinsic capacity of the foetus for growth and the maternal environment. Lambs born in multi-foetus litters have relatively small placentae with fewer cotelydons, and lower birth weights. Litter-size-dependent intrauterine growth restriction (IUGR) is evident at mid gestation when metabolic needs of the conceptus are moderate, and overnutrition of ewes with multiple foetuses does not promote growth of their foetuses to the size of singletons. Those observations suggest that placental and conceptus growth in multi-foetus pregnancies is reprogrammed at mid gestation by an as yet undefined mechanism to attenuate foetal growth. This may protect the foetus from severe nutritional insult during late gestation, when its daily growth rate is at a maximum. In that way, lambs born in large litters with relatively lower birth weights may not experience the long-term physiological insults that can be observed in small lambs born to undernourished ewes.

The opinions of seven respondent groups about the relative importance of different practices pertaining to the welfare of Australian beef cattle, sheep and goats were surveyed. Respondent groups comprised farmers, livestock transportation representatives, veterinarians, meat processors, animal welfare advocates, animal welfare scientists and government officers. The survey consisted of a web-based adaptive conjoint analysis questionnaire, which was administered to a sample population that was selected randomly for large respondent groups and comprehensively for small groups. The hierarchy of opinion concerning the importance of the different beef cattle practices was: stockmanship > ground (road and rail) transport > spaying > food supply > dehorning > stunning > shelter > identification > pretransport food and water deprivation > castration > sea transport > mustering > confinement. For sheep/goat practices the hierarchy was: parasite control > mulesing > shelter > stockmanship > tail docking > ground transport > feeding > predation > stunning > castration > pretransport food and water deprivation > sea transport > mustering. The method of performing invasive procedures was perceived as less important than the provision of pain relief. Differences in opinion were evident between respondent groups, with animal welfare advocates tending to focus on painful procedures more than those with direct involvement in the industry.

Because of an increasing demand for animal-source foods, an increasing desire to reduce poverty and an increasing need to reduce the environmental impact of livestock production, tropical farming systems with livestock must increase their productivity. An important share of the global human and livestock populations are found within smallholder mixed-crop–livestock systems, which should, therefore, contribute significantly towards this increase in livestock production. The present paper argues that increased livestock production in smallholder mixed-crop–livestock systems faces many constraints at the level of the farm and the value chain. The present paper aims to describe and explain the impact of increased production from the farm and farmers’ perspective, in order to understand the constraints for increased livestock production. A framework is presented that links farming systems to livestock value chains. It is concluded that farming systems that pass from subsistence to commercial livestock production will: (1) shift from rural to urban markets; (2) become part of a different value chain (with lower prices, higher demands for product quality and increased competition from peri-urban producers and imports); and (3) have to face changes in within-farm mechanisms and crop–livestock relationships. A model study showed that feed limitation, which is common in tropical farming systems with livestock, implies that maximum herd output is achieved with small herd sizes, leaving low-quality feeds unutilised. Maximal herd output is not achieved at maximal individual animal output. Having more animals than required for optimal production – which is often the case as a larger herd size supports non-production functions of livestock, such as manure production, draught, traction and capital storage – goes at the expense of animal-source food output. Improving low-quality feeds by treatment allows keeping more animals while maintaining the same level of production. Ruminant methane emission per kg of milk produced is mainly determined by the level of milk production per cow. Part of the methane emissions, however, should be attributed to the non-production functions of ruminants. It was concluded that understanding the farm and farmers’ perceptions of increased production helps with the understanding of productivity increase constraints and adds information to that reported in the literature at the level of technology, markets and institutions.

The fatty acid composition of sperm affects the fertilization rate. The objective was to investigate the effects of dietary fish oil (as a source of n-3 fatty acids) on semen quality and sperm fatty acid composition in sheep. Eight Zandi fat-tailed rams were randomly allocated into two groups and fed either a control diet or a diet supplemented with fish oil. Both diets were isocaloric and isonitrogenous and were fed for 13 weeks, starting in the middle of the breeding season. Semen samples were collected weekly and their characteristics evaluated by standard methods, whereas samples collected at the start and end of the study were assessed (gas chromatography) for sperm lipid composition. Mean (±s.e.m.) sperm concentrations (4.3 × 109 ± 1.3 × 108v. 3.9 × 109 ± 1.3 × 108 sperm/ml and percentages of motile (77.25 ± 3.34 v. 60.8 ± 3.34) and progressively motile sperm (74.13 ± 1.69 v. 62.69 ± 1.69) were significantly higher in the fish oil group than control. Dietary fish oil increased the proportion of docosahexaenoic acid (DHA, C22:6 n-3) in sperm fatty acid composition. We concluded that feeding fish oil as a source of n-3 fatty acids attenuated seasonal declines in semen quality in rams, perhaps through increased DHA in sperm.

Cell-based meat, also called ‘clean’, lab, synthetic or in vitro meat, has attracted much media interest recently. Consumer demand for cellular meat production derives principally from concerns over environment and animal welfare, while secondary considerations include consumer and public health aspects of animal production, and food security. The present limitations to cellular meat production include the identification of immortal cell lines, availability of cost-effective, bovine-serum-free growth medium for cell proliferation and maturation, scaffold materials for cell growth, scaling up to an industrial level, regulatory and labelling issues and at what stage mixing of myo-, adipo- and even fibrocytes can potentially occur. Consumer perceptions that cell-based meat production will result in improvements to animal welfare and the environment have been challenged, with the outcome needing to wait until the processes used in cell-based meat are close to a commercial reality. Challenges for cell-based meat products include the simulation of nutritional attributes, texture, flavour and mouthfeel of animal-derived meat products. There is some question over whether consumers will accept the technology, but likely there will be acceptance of cell-based meat products, in particular market segments. Currently, the cost of growth media, industry scale-up of specific components of the cell culture process, intellectual property sharing issues and regulatory hurdles mean that it will likely require an extended period for cellular meat to be consistently available in high-end restaurants and even longer to be available for the mass market. The progress in plant-based meat analogues is already well achieved, with products such as the ImpossibleTM Burger and other products already available. These developments may make the development of cellular meat products obsolete. But the challenges remain of mimicking not only the nutritional attributes, flavour, shape and structure of real meat, but also the changes in regulation and labelling.

There is currently a lack of research investigating the effectiveness of commercial broiler enrichments, and in particular the ability of these additions to create opportunities for positive welfare. One aim of this study was to investigate whether offering broiler chickens enrichments that have recently been found to be preferred leads to increased levels of activity. A second aim was to investigate the emotional effects of provision of these enrichments by assessing levels of fearfulness and play-like activity. Commercially housed broilers were assigned to treatment houses containing either: (1) platform perches, (2) platform perches+peat dust baths, (3) no enrichment (control). Activity levels and play behaviours in unenriched areas of the house were measured in weeks 3, 4 and 5. Levels of active behaviours, such as foraging and locomotion, were determined from video recordings of undisturbed birds in unenriched areas of the house. To stimulate play-like behaviours an observer walked through the birds, displacing them and creating a space. The broilers using the space were then filmed for 5 min and the occurrences of frolicking, sparring and food-running were recorded. Fearfulness of broilers in unenriched areas of the house, and also when using enrichments, was measured using observer avoidance tests in week 5. We found that creating space among the broilers was a successful method of stimulating play (largely sparring and frolicking), with play being observed in 93% of videos, however the presence of enrichments did not have an effect on the level of play recorded (P>0.05). There was also no treatment effect on activity levels of broilers in unenriched areas (P>0.05), however levels of overall activity decreased as broilers aged. Compared with the control, flight distances in unenriched areas were significantly lower in the perches+dust bath treatment (P=0.026), and were numerically lower in the perches treatment. This suggests a reduction in fearfulness with increased environmental complexity, and thus possible welfare benefits. It is suggested that further research should investigate whether increasing the level of provision of these enrichments leads to more marked improvements in welfare.

Modern fast-growing broilers spend excessive periods resting and their activity further decreases with age. Inactivity has been suggested to increase impaired gait and the incidence of leg disorders. Tibial dyschondroplasia (TD) is a common leg pathology in broilers. A more complex environment might facilitate more activity and improve leg health. Perches or elevated platforms bring variety to broilers’ environment and could motivate more locomotion. This study examined the impact of perches and elevated platforms on walking ability, the occurrence of TD and level of bone ash and mineral contents. The investigation was performed on four commercial broiler farms throughout six consecutive batches with platforms and four to five with perches. On each farm at least two separate houses were included, enabling the comparison of furnished flocks to control flocks during each batch. Plastic slats with ramp access elevated by 30 cm or wooden perches of 10 and 30 cm height were offered in the furnished house. Farmers recorded the platform and perch usage twice a week with a five-point scale. Gait was scored before slaughter on a six-point scale according to the Welfare Quality® assessment protocol for poultry. The severity of TD was determined using a four-point scale on farm from all birds gait scored as 3 and at slaughter from 200 birds/flock. Farmers estimated 50% to 100% of the platforms to be occupied in all flocks throughout the entire growing period. Only single birds were perching, thus perch structures were constantly evaluated to be empty. Due to the low use, the perch-equipped houses were excluded when analysing bone content, walking ability and TD. On average, 30% of the tested birds exhibited gait score ⩾3. Younger scoring age resulted in a lower mean gait score and a lower percentage of scores 3 and 4 to 5. Overall, 2.3% of the birds examined at slaughter and 3.5% of the birds with gait score 3 were affected by TD. Leg health was better in birds with access to platforms: mean gait score, the percentage of birds scoring 3, and TD percentage and severity were lower in birds in platform-equipped houses. Elevated structures such as platforms, offering additional possibilities for locomotion to broilers seem to improve their leg health.

Muscle chemical analysis and muscle identification both were attempted by using visible and near infrared reflectance spectroscopy (NIRS). Seventy-eight beef muscles (m. longissimus dorsi) from Hereford cattle were used. The samples were scanned in a NIRS monochromator instrument (NIRSystems 6500, Silver Spring, MD, USA) in reflectance mode (log 1/R). Both intact and minced muscle presentation to the instrument were explored. Predictive equations were made using ISI software (Infrasoft International, Port Matilda, PA, USA) and muscle identification was performed by Principal Component Analysis (PCA) and Soft Independent Modelling of Class Analogy (SIMCA). The coefficient of determination in calibration (R2CAL) and standard error in cross validation (SECV) for the intact sample presentation were 009 (SECV: 15·6), 0·89 (SECV: 46·9), 0·48 (SECV: 23·9) for moisture (M), fat and crude protein (CP) on g/kg fresh weight basis respectively. R2CAL and SECV for minced sample presentation were 0·41 (SECV: 161), 0·92 (SECV: 43·4), 0·71 (SECV: 20·5) for M, fat and CP on g/kg fresh weight basis respectively. Qualitative analysis of optical information through PCA and SIMCA analysis showed differences in muscles resulting from two different feeding systems.

The voluntary intake and digestibility of four hays were measured with eight sheep using two 4 × 4 Latin squares. Measurements were made during the last 8 days of each 3-week period. The degradation characteristics of the hays were measured by incubating samples (in nylon bags) for 12, 24, 48 or 72 h in the rumen of four sheep fitted with rumen cannulae and given a good hay. The exponential p = a + b(1 − e−α) where p = degradation loss, t = time, and a, b and c are constants, was fitted. The potential degradability (defined as a + b) of the dry matter (DM) of the four hays was 0·76, 0·66, 0·54 and 0·46 with corresponding voluntary intakes of 71, 62, 52 and 45 ± 2·9 g DM per kg M075 per day. The in vivo digestibilities were 0·61, 0·59, 0·46 and 0·45 (s.e. 0·013) respectively and corresponded to 23-, 25-, 31- and 67-h degradation. Voluntary intake was better related to potential degradability (and degradability at 12, 24, 48 and 72 h) than to in vivo digestibility. It is concluded that the degradation characteristics of forages may have useful application in predicting voluntary intakes, and that potential degradability could be used to define the rumen (jegradable nitrogen content necessary with any particular forage.

Insulin-like growth factors (IGFs) and their binding proteins play an essential role in regulating animal growth and metabolism. The initial portion of the current review focuses on the physiological effects of the IGFs and delineates their role as regulators of animal growth and metabolism. The role of IGFs as mediators of growth hormone effects, as insulin-like metabolic regulators and as foetal growth regulators is discussed. The remainder of the review is devoted to the IGF binding proteins, their modulation of IGF action and their role in foetal and postnatal regulation of growth.

Even though extensive research has examined the role of nutrition on milk fat composition, there is less information on the impact of forages on milk fatty acid (FA) composition. In the current study, the effect of replacing grass silage (GS) with maize silage (MS) as part of a total mixed ration on animal performance and milk FA composition was examined using eight multiparous mid-lactation cows in a replicated 4 × 4 Latin square with 28-day experimental periods. Four treatments comprised the stepwise replacement of GS with MS (0, 160, 334 and 500 g/kg dry matter (DM)) in diets containing a 54 : 46 forage : concentrate ratio on a DM basis. Replacing GS with MS increased (P < 0.001) the DM intake, milk yield and milk protein content. Incremental replacement of GS with MS in the diet enhanced linearly (P < 0.001) the proportions of 6:0–14:0, decreased (P < 0.01) the 16:0 concentrations, but had no effect on the total milk fat saturated fatty acid content. Inclusion of MS altered the distribution of trans-18:1 isomers and enhanced (P < 0.05) total trans monounsaturated fatty acid and total conjugated linoleic acid content. Milk total n-3 polyunsaturated fatty acid (PUFA) content decreased with higher amounts of MS in the diet and n-6 PUFA concentration increased, leading to an elevated n-6 : n-3 PUFA ratio. Despite some beneficial changes associated with the replacement of GS with MS, the overall effects on milk FA composition would not be expected to substantially improve long-term human health. However, the role of forages on milk fat composition must also be balanced against the increases in total milk and protein yield on diets containing higher proportions of MS.

‘Welfare is a wide term that embraces both the physical and mental wellbeing of the animal. Any attempt to evaluate welfare, therefore, must take into account the scientific evidence available concerning the feelings of animals that can be derived from their structure and functions and also from their behaviour.’ So stated the Report prepared by the Technical Committee to Enquire into the Welfare of Animals kept under Intensive Livestock Husbandry Systems under the chairmanship of Professor Rogers Brambell (Command Paper 2836, 1965).

Lameness in sows is an emerging disease condition with major effects on animal welfare and economics. Yet the direct impact on reproduction results remains unclear. The present field study investigated the impact of lameness and claw lesions throughout the reproductive cycle on (re)production results of sows. In five farms, a total of 491 group-housed sows were followed up for a period of one reproductive cycle. Sows were assessed for lameness every time they were moved to another area in the farm. Claw lesions were scored at the beginning and at the end of the cycle. Reproduction results included the number of live-born piglets, stillborn piglets, mummified fetuses and crushed piglets, weaning-to-oestrus interval and the presence of sows not showing oestrus post weaning, returning to service and aborting. Sows that left the group were recorded and the reason was noted. A mean prevalence of lameness of 5.9% was found, although it depended on the time in the productive cycle. The highest percentage of lame sows (8.1%) was found when sows were moved from the post-weaning to the gestation stable. No significant associations were found between lameness and reproduction parameters with the exception of the effect on mummified foetuses. Wall cracks, white line lesions, heel lesions and skin lesions did have an effect on farrowing performance. Of all sows, 22% left the group throughout the study, and almost half of these sows were removed from the farm. Lameness was the second most important reason for culling. Sows culled because of lameness were significantly younger compared with sows culled for other reasons (parity: 2.6 ± 1.3v. 4.0 ± 1.8). In conclusion, the present results indicate that lameness mainly affects farm productivity indirectly through its effect on sow longevity, whereas claw lesions directly affect some reproductive parameters. The high percentage of lame sows in the insemination stable indicate that risk factor studies should not only focus on the gestation stable, but also on housing conditions in the insemination stable.

Fatty acid synthase (FASN) is known as a crucial enzyme of cellular de novo fatty acid synthesis in mammary gland which has been proved as the main source of short and medium-chain fatty acids of milk. However, the regulatory role of FASN in goat-specific milk fatty acids composition remains unclear. We cloned and analyzed the full-length of FASN gene from the mammary gland of Capra hircus (Xinong Saanen dairy goat) (DQ 915966). Comparative gene expression analysis suggested that FASN is predominantly expressed in fat, small intestine and mammary gland tissues, and expresses higher level at lactation period. Inhibition of FASN activity by different concentrations (0, 5, 15, 25 and 35 μM) of orlistat, a natural inhibitor of FASN, resulted in decreased expression of acetyl-CoA carboxylase α (ACCα), lipoprotein lipase and heart-type fatty acid binding protein (H-FABP) in a concentration-dependent manner in goat mammary gland epithelial cells (GMEC). Similar results were also obtained by silencing of FASN. Additionally, reduction of FASN expression also led to apparent decline of the relative content of decanoic acid (C10:0) and lauric acid (C12:0) in GMEC. Our study provides a direct evidence for inhibition of FASN reduces cellular medium-chain fatty acids synthesis in GMEC.

The objective of this study was to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for porcine salivary chromogranin A (CgA) measurements, using a species-specific antibody, and evaluate its behaviour in an acute stress model. Polyclonal antibodies were produced in rabbits immunized with a synthetic porcine fragment of CgA359−379 and used to develop a sandwich TR-IFMA. This TR-IFMA was analytically validated and showed intra- and inter-assay coefficients of variation of 6.23% and 5.82%, respectively, an analytical limit of detection of 4.27 × 10−3 μg/ml and a limit of quantification of 24.5 × 10−3 μg/ml. The assay also demonstrated a high level of accuracy, as determined by linearity under dilution (r = 0.975) and recovery tests. When a model of experimental acute stress, in which animals were immobilized for 3 min with a nose snare (stressor stimulus), was applied, a significant increase (P < 0.05) in CgA levels in saliva was detected at 15 min post-stressor stimulus. These results indicate that the assay developed in this study could measure CgA in porcine saliva in a reliable way and that the concentrations of CgA in saliva samples of pigs increase after an acute stress situation.