Non-invasive methods for analysing pig welfare biomarkers.

At present, the assessment of pig welfare quality has gained significant importance, prompting the exploration of novel biomarkers for this purpose. Traditionally, these biomarkers have been monitored in the blood; however, blood sampling is considered an invasive procedure. Currently, non-invasive methods for collecting samples are emerging as viable alternatives for assessing these biomarkers. This article aims to present the current knowledge regarding the use of non-invasive methods for analysing pig welfare biomarkers, specifically focusing on the saliva, hair, faeces, and urine as matrices to determine these biomarkers. The saliva analysis encompasses various biomarkers, such as cortisol, alpha-amylase, chromogranin A, the total esterase, oxytocin, acute phase proteins, adenosine deaminase, immunoglobulins and parameters of redox homeostasis. Cortisol, a specific biomarker, can be determined in the hair, urine and faeces, while urine samples allow for the analysis of catecholamines as non-invasive markers of pig welfare.

Assessing Substrate Utilization and Bioconversion Efficiency of Black Soldier Fly (Hermetia illucens) Larvae: Effect of Diet Composition on Growth and Development Temperature.

Black soldier fly larvae (BSFL) can utilize food by-products or residues for growth, benefiting farm animal's diets' production sustainability. The experiment aimed to assess the effect of different substrate compositions on larval growth, chemical composition, and substrate temperature. BSFL were allocated to one of the four diets (control, vegetable, carnivorous, and omnivorous) for the entire experiment (8 days). The temperature was measured twice daily using a thermal-imaging camera, and the accumulated degree hours (ADH) was calculated. The results showed that the larvae fed the vegetable diet exhibited a significantly reduced growth performance, with a biomass reduction of 26.3% compared to the control diet; furthermore, vegetable-fed larvae showed a lower dry matter content (-30% compared to the average of other diets) due to lower fat content (-65% compared to average of other diets). The nutritional composition of larvae fed an omnivorous diet was similar to larvae fed a high-quality substrate diet (control diet-chicken feed), indicating that the omnivorous diet could be an ideal solution for rearing BSFL larvae; however, the current European legislation prohibits the use of animal meal. The study also revealed that substrate temperatures did not have a discernible influence on larval growth, further emphasizing the importance of diet in BSFL rearing strategies.

Content of Toxic Elements (Arsenic, Cadmium, Mercury, Lead) in Eggs from an Ethically Managed Laying Hen Farm.

Domestic chicken farming has been promoted and spread in several Italian municipalities and worldwide as an aid to the self-consumption of domestically produced food. This study investigated the levels of four toxic elements (As, Cd, Hg, and Pb) in eggs from an ethical laying hen farm, comparing the element concentrations with those possibly present in supermarket eggs. A total of 201 eggs, 141 from the farm and produced by different hen genotypes, and 60 from the supermarket, were collected. The levels of the toxic elements were evaluated in the yolk, albumen, and eggshells of all eggs. The results show that the supermarket eggs' yolk and albumen were more contaminated with lead, compared to the rural eggs. Contrarily, the mean content of arsenic was higher in the albumen and eggshells of the rural eggs, compared to the supermarket eggs. The cadmium content was below the LOQ (0.005 mg/kg) in all samples. The mercury content was below or around the LOQ in all rural eggs. Overall, the supermarket egg albumens were significantly more contaminated than the rural ones. No significant differences were found in quality parameters for both types of eggs. The toxic element values that were detected were in line with other studies in the literature. However, despite the concentrations found not representing a risk to the consumers' health, the results of this study raise a potential food safety issue, and it would be desirable to set specific MRLs for eggs for consumers' protection.

Effects of Lavender Essential Oil Inhalation on the Welfare and Meat Quality of Fattening Heavy Pigs Intended for Parma Ham Production.

We assessed the effects of inhalation administration of lavender essential oil (LEO) either once (L1) or twice (L2) a day on animal welfare indicators, carcass and meat quality of Italian heavy pigs. Pigs (n = 108) were allotted to three experimental groups (control -C-, L1 and L2) and lavender was administered, via a vaporizer device, to the treated groups during the entire fattening-finishing period (79-160 kg BW). Tail lesion severity was reduced in L1 at the end of the trial compared to the other groups (p < 0.05). Body lesion counts, however, were higher in L2 than in C (p < 0.05), resulting in a more severe overall damage classification (p < 0.01). At slaughter, no differences were observed in carcass traits or blood stress indicators, only minor differences were observed in meat quality, and no LEO residual was found in fat or lean tissues, highlighting the preserved suitability of thighs for the dry curing process. While it was not possible to conclude on the ability to improve animal welfare of vaporized LEO in this production phase, the absence of adverse effects on meat quality and the discrepancies observed regarding the body lesions in L1 and L2 make further studies on behavioral aspects and the method of administration (route, frequency) of the product desirable.

Growth Performance and Feed Intake Assessment of Italian Holstein Calves Fed a Hay-Based Total Mixed Ration: Preliminary Steps towards a Prediction Model.

The aim of this study was to evaluate the effects of a complete hay-based total mixed ration (TMR) for calves, focusing on their feed intake, animal growth performance, and fecal output, and to develop a preliminary estimation equation for solid feed intake and body weight in Holstein heifer calves. Twenty female Italian Holstein calves (37.14 ± 2.72 kg) born between February and July were studied from the day of birth until 77 days of age. From the fourth day of life, they were fed 3 L/day of pasteurized milk twice daily and supplemented with the same hay-based TMR. The data on feed intake, fecal characteristics, and growth performances were collected and showed that these calves had adequate parameters. Moreover, the data collected was used to create equations to predict body weight and solid feed intake using a mixed model. The goodness of fit of the developed equations was evaluated by coefficients of determinations (R2). The equation obtained shows high R2 (0.98 for solid feed intake and 0.99 for calf weight), indicating the satisfactory precision and accuracy needed to predict female calves' body weight and solid feed intake.

Mortality during Transport of Pigs Subjected to Long Journeys: A Study in a Large European Abattoir.

To date, especially in Europe, few studies have analysed the implications of long journeys on pig welfare and economic losses, expressed in terms of transport mortality. This study retrospectively analysed data collected from slaughtering registers and travel journals in a large Italian abattoir. We focused on pig transports coming from abroad and arriving at the slaughter plant after long journeys (a total of 59,982 pigs over 370 journeys). We explored the relationship between mortality and the following variables: country of origin, journey duration, astronomical season, stocking density on the truck, number of stops, and prolonged stops during the journey (lasting more than 60 min, likely due to traffic jams or truck problems). Overall, the low mortality rate observed (0.09%) was in line with European estimates. The factors with a significant or tendential effect on mortality during transport were the astronomical season (p = 0.0472, with higher mortality in spring) and the presence of prolonged stops during the journey (p = 0.069, tendential effect). Journey duration, stocking density, country of origin, and the number of stops were not statistically significant. In conclusion, based on this case study, using transport mortality combined with data collected during the common routine activity by the Public Veterinary Services in slaughterhouses could be a simple screening method for identifying problematic journeys or transport conditions.

Influence of Increased Freedom of Movement on Welfare and Egg Laying Pattern of Hens Kept in Aviaries.

This work investigates the effects of structural modifications on the welfare level and laying patterns of hens in a three-tier commercial aviary system. Four experimental groups were used: C (control, housed in a traditional aviary); LM (longitudinal movement, in which internal partitions were removed); VM (vertical movement, in which ramps were installed); and FM (freedom of movement, both LM and VM modifications). Hens showed worse body condition scores (p < 0.05) in all the modified aviaries, while plumage condition was improved in FM but worsened in VM (p < 0.05). No significant effect was observed on egg deposition patterns, egg quality or keel bone damage. When ramps were available (VM and FM groups), hens reduced the number of flights and increased the number of walks from 0.52 to 7.7% of the displacements on average (p < 0.05). Apart from some feather pecking concerns in VM (likely due to overcrowding in some favourite aviary areas), LM and FM seemed to facilitate animal movement and promote species−specific behaviour. It is concluded that hen welfare in aviary systems can be improved by means of tailored structural modifications. Producers may therefore adopt some of these modifications (providing ramps and/or removing vertical barriers) to enhance the welfare of hens.

Literature Review on the Pre-Slaughter Welfare of Italian Heavy Pigs.

This work provides a narrative review of the available information on the welfare of Italian heavy pigs in the pre-slaughter phase (transport, lairage, and stunning). The meat from these pigs is used for specific PDO (Protected Designation of Origin) products, and the production rules for these specialties require higher body weight (160-170 kg) and age (in general more than 9 months) at slaughter than in most other countries. This may lead to specific behavioral and physiological needs of pigs. The present paper summarizes the main research findings and knowledge gaps for each of the pre-slaughter phases. Studies are presented according to the four principles of the Welfare Quality assessment protocol (good feeding, good housing, good health, and appropriate behavior). The results of the literature review indicate a lack of knowledge on several aspects. Most of studies were carried out in a single slaughterhouse, making it difficult to identify risk factors and confounding effects. Moreover, animal-based measures were assessed using different protocols, reducing the possibility of comparison across studies. These findings may serve as a basis for the development of specific research studies and policies aimed at enhancing the animal welfare level and the ethical attributes of this renowned production, also in accordance with consumers' expectations.

Update on animal welfare perception by Italian consumers: A descriptive survey.

The aim of this study was to evaluate if and how the perception of animal welfare by Italian consumers and their purchasing behavior of animal-friendly foods have evolved in recent years. The research was carried out through a survey on a representative sample of Italian consumers (n=969). Responses showed that 69% of consumers declared to pay attention to animal welfare at time of purchasing, and they mostly acquired information about animal welfare through the mass media. In accordance with previous surveys, the categories perceived as having the highest welfare level were fish and dairy cows, whereas the lowest welfare level was attributed to broilers and pigs. Compared to previous surveys, Italian consumers seem to have improved their knowledge about the legislation on animal protection and increased their interest toward animal welfare issues. However, they still find it difficult to identify animalfriendly labels, together with a lack of knowledge about non-conventional (e.g. extensive and organic) farming systems.

Knowledge and Perspectives on the Welfare of Italian Heavy Pigs on Farms.

This review aims to give an overview of the most significant available information on welfare issues of Italian heavy pigs on farms. These animals, whose meat is used to produce typical products, are characterized by much higher body weights and age at slaughter (approximately 160-170 kg, and 9 months of age) than most pigs reared in Europe, resulting in peculiar behavioral and rearing-related needs. To highlight the main findings and the areas in which additional research is needed, the paper was organized by allocating the studies on the basis of the Welfare Quality four principles framework, i.e., good feeding, good housing, good health, and appropriate behavior. The results indicate the possibility of improvement with regard to the management of feeding, watering, and some environmental parameters (light, thermal comfort, enrichments) and a lack of knowledge on the actual space requirements (and a corresponding regulatory gap). Moreover, deficiencies concerning the prevalence of injuries and disease and the relationship between lesions observed post-mortem and rearing conditions needs to be addressed. Lastly, the absence of research concerning the evaluation of the emotional state of animals has been highlighted. It is hoped that these findings will, in the future, serve as a basis for the development of specific policies for these animals in order to increase the ethical value of the entire production chain, in accordance with consumers' demand and expectation for high animal welfare standards.

Pre-Slaughter Sources of Fresh Meat Quality Variation: The Case of Heavy Pigs Intended for Protected Designation of Origin Products.

This study focused on loin quality in Italian heavy pigs intended for the production of PDOs (Protected Designation of Origin) products, and investigated the pre-slaughter factors which negatively affect the quality of fresh meat. Data were collected on 44 shipments (loads) of pigs. Shipments were carried out under commercial conditions. Several pre-slaughter parameters were recorded within the entire process (on-farm, during transport, and at the slaughterhouse). On a subset of pigs (10 animals from every load, N = 440), serum cortisol and creatine kinase were measured and loin samples were analyzed for pH, instrumental color, drip loss, cooking loss, shear force, and sensory quality. Cluster analysis of the instrumentally-assessed meat quality parameters allowed the categorization of the shipments into two clusters: lower quality (LQ) and higher quality (HQ). Our results showed that the factors with significant differences between the two clusters were journey duration, ambient temperature, distance traveled, and irregular behaviors (slipping, falling, and overlapping) at unloading (all greater in LQ, p < 0.05). The pre-slaughter conditions associated with lower loin quality were ambient temperatures above 22 °C, distance traveled above 26 km, travel duration between 38-66 min, more than 5.9% of animals showing irregular behaviors at unloading.

Health Risk Perception, Consumption Intention, and Willingness to Pay for Pig Products Obtained by Immunocastration.

Surgical castration without the use of anaesthesia and/or analgesia is considered to be detrimental for the welfare of pigs and for this reason its abandonment is advocated. Immunocastration is a more welfare-friendly alternative method; however, stakeholders in the pork sector fear consumer rejection due to perceived safety issues of immunocastrated meat. This work aimed to analyse whether Italian consumers perceive a health risk arising from the use of this technique and, if so, how the perceived risk may influence the purchase choices and the willingness to pay for products derived from immunocastrated animals. To achieve this objective, a survey was carried out on a representative sample of the Italian population. The results highlight that consumers perceive different levels of risk related to the use of immunocastration and that this influences purchasing behaviour and willingness to pay. Moreover, it should be noted that the willingness to pay is also influenced by certain demographic factors, since this is positively associated with younger respondents with lower incomes and less knowledge of farming systems, who live in rural areas and have a greater sensitivity to animal welfare. Given the concerns expressed by consumers, particular attention must be paid to the information transmitted if this technology will be widely implemented in pig husbandry.

Identification of Possible Pre-Slaughter Indicators to Predict Stress and Meat Quality: A Study on Heavy Pigs.

This study aimed at identifying possible pre-slaughter indicators and/or indexes to be used to predict pig stress response and meat quality variation. Data were collected on 44 shipments (loads) of Italian heavy pigs. For each shipment, several pre-slaughter parameters were recorded on farm, during transport, and at the slaughterhouse. Blood and meat samples were taken from 10 pigs from every of the 44 loads included in the study (N = 440). Blood samples were used to assess cortisol and creatine kinase levels, whereas meat samples were used to assess meat quality (pH, instrumental color, tenderness, water-holding capacity, and sensory analysis). Cluster analysis of blood parameters allowed the categorization of the shipments into two main clusters: Lower Stress (LS) and Higher Stress (HS). The variables/indexes statistically differing between the two clusters were: average vehicle speed during transport, welfare index at slaughter (i.e., "slaughter score"), overall transport and slaughter welfare index (TSWI), distance travelled, and behaviors (slips, falls, overlaps) during unloading, which appeared to be the best descriptors of the welfare conditions experienced by Italian heavy pigs during pre-slaughter handling. No consistent effects of the stress level experienced on meat quality was detected, which warrants the need for further studies conducted under more variable pre-slaughter conditions, with the aim of simplifying and improving the TSWI.

Effect of dietary inclusion of different lipid supplements on performance and carcass quality traits of growing beef heifers.

The aim of this study was to evaluate the impact of dietary extruded flaxseed and/or rumen-protected lipids on growth performance and carcass quality of growing beef heifers. Sixty-three crossbreed heifers (Charolais X Limousine) were distributed into seven experimental groups, balanced in terms of age and live weight. Diets fed to the groups were isoproteic and differed in both, the dietary lipid source (extruded linseed and/or rumen-protected conjugated linoleic acid) and the supplementation length (90 or 180 days before slaughtering), having the same total amount of lipids and vitamin E, during their finishing period. The results obtained in the present study confirm that in low-protein diets, the inclusion of rumen-protected CLA, alone or in combination with flaxseed, did not bring any evident effect on feed intake, performance and carcass quality traits of growing beef heifers.

Towards the Abandonment of Surgical Castration in Pigs: How is Immunocastration Perceived by Italian Consumers?

Immunocastration of pigs represents an alternative method to surgical castration, being more respectful of animal welfare. However, this new technology may not be accepted by consumers due to their perception of possible risks tied to the use of the product, thus representing a concern for the production sector. The study aimed at verifying the attitude of Italian consumers towards immunocastration and to assess whether their perception can be affected by science-based information on advantages and disadvantages of immunocastration. A total of 969 consumers (divided in three groups representative of the Italian population) were contacted and asked to complete an online questionnaire. Only technical (neutral) information on immunocastration was provided to the first group; the second and the third group received information on the advantages (+) and disadvantages (-) of the technique, shown in reverse order (+/- and -/+, respectively). The level of information did not affect consumers' perception of immunocastration. Overall, immunocastration is perceived in a predominantly positive manner (54.5%), with a relatively low level of risk perception (34.2%), and a good willingness to pay more for meat deriving from immunocastrated pigs (+18.7%).

Partially defatted black soldier fly larva meal inclusion in piglet diets: effects on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features.

BACKGROUND: The aim of this trial was to investigate the effects of different inclusion levels of a partially defatted black soldier fly (BSF, Hermetia illucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individually weighed (initial body weight (IBW): 6.1 ± 0.16 kg) and randomly allocated to 3 dietary treatments (4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels (0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I (from d 1 to d 23) and II (from d 24 to d 61). The weight gain (WG), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations. RESULTS: No overall significant differences were observed for growth performance (P > 0.05), except for the ADFI of phase II, which showed a linear response to increasing BSF meal levels (P < 0.05, maximum for the BSF10 group). Dietary BSF meal inclusion did not significantly influence the blood profile, except as far as monocytes and neutrophils are concerned, and these showed a linear and quadratic response, respectively, to increasing BSF meal levels (P < 0.05, maximum for the BSF10 and BSF5 groups, respectively). On the other hand, the nutrient digestibility, gut morphology and histological features were not affected by dietary BSF meal inclusion (P > 0.05). CONCLUSIONS: The obtained results show that a partially defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.

Effects of increased space allowance on animal welfare, meat and ham quality of heavy pigs slaughtered at 160Kg.

Sixty barrows (Body Weight-BW- range: 23.9-160 kg) were allotted to two experimental groups (6 pens of 5 pigs each): the control group was kept at a space allowance of 1m2/head; the second group was kept at 1.3m2/head. Behaviour, growth parameters, carcass and meat quality were assessed, as well as fat and cured ham quality. Results showed that pigs raised at 1.3m2/head spent more time laying (particularly in lateral recumbency, P<0.01 and P<0.001, respectively) compared to pigs kept at lower space allowance. They also reduced the aimless exploration of the slatted pen floor (P<0.001) and increased overall expression of other, mainly active, behaviors (e.g., drinking, walking and standing, P<0.01). Pigs raised at 1.3m2/head showed higher final BW (P = 0.02), more favourable Average Daily Gain (ADG) and gain-to-Feed ratio (G:F) both during the last period of the trial (P<0.05 for both parameters) and over the entire trial (P = 0.01 for both parameters). No significant difference was observed between groups for carcass traits and the main meat quality attributes. Subcutaneous fat from green hams had higher α-linolenic acid content (P<0.01) in the group reared at greater space allowance. Green hams from this group lost less weight at trimming (P<0.01) and the resulting cured hams received better sensory evaluations (P<0.05). No difference was observed in fatty acid composition and unsaturation levels of the subcutaneous fat from cured hams. Our data suggest that heavy pigs intended for Parma ham would benefit from the adoption of higher individual floor space allowances, both in terms of animal welfare (increased possibility to rest) and of productive parameters, without having any detrimental effect on the suitability of the thighs for dry-curing or on the quality of the final product.

Hair cortisol determination in sows in two consecutive reproductive cycles.

Hair analysis has been proposed as a minimally invasive technique capable of furnishing information regarding the stress response during medium- and long-term periods. Bristle samples were collected from the rump region of sows at three key physiological phases (before delivery - BD; weaning time - WT; pregnancy diagnosis - PD) during consecutive reproductive cycles in order to test swine hair as a reliable matrix of cortisol evaluation. Cortisol was extracted from the bristles and assayed using radioimmunoassay. The highest mean hair cortisol concentrations were demonstrated (p<0.001) at the PD time points (20.1±.95 and 16.29±2.15 pg/mg). Moreover, cortisol was significantly higher (p<0.001) at BD2 (10.48±0.96 pg/mg) as compared to BD1 (5.17±0.51 pg/mg) and WT1 (6.01±0.47 pg/mg). The various physiological phases had a significant effect on cortisol concentration (p<0.00001) with a higher cortisol concentration found during late pregnancy and lactation than in early-mid pregnancy. This could be due not only to the physiological hormonal status, but also to the different housing conditions (single crates vs. group housing). The season of the year was also observed to have an effect (p<0.005), with the lowest cortisol concentration recorded during the hot season.

Meat and ham quality of Italian heavy pigs subjected to different illumination regimes.

In order to attain a good level of welfare pigs require a sufficient environmental illumination.Therefore minimum levels for light duration and light intensity are set up by the European legislation.Two independent and separate trials were aimed to determine whether an increase above the minimum mandatory levels of lighting duration (14 vs. 8 hours/day at 70 lux: trial A) or of light intensity (80 vs. 40 lux over a 12-hour lighting period: trial B) could modify the carcass traits and meat and cured hams quality of Italian heavy pigs (body weight at slaughtering of about 160 kg). Slaughtering parameters, fresh meat quality and fatty acid composition of raw thighs and cured hams were assessed. Pigs receiving the longer photoperiod showed a tendency (P < 0.1) toward a higher slaughtering body weight and a higher saturation degree (P < 0.01) of subcutaneous fat of the raw thighs, while cured hams deriving from pigs subjected to the higher illumination intensity showed a higher (P < 0.05) relative content of polyunsaturated fatty acids. Our results indicate that, within a moderate range of light intensity and given an appropriate dark period for animal rest, neither a prolonged photoperiod nor a higher light intensity caused any negative effect on the carcass traits, meat or quality of long-cured hams.

Oxidative stability of pork meat lipids as related to high-oleic sunflower oil and vitamin E diet supplementation and storage conditions.

The aim of this research work was to evaluate the oxidative stability of pork meat lipids as related to dietary supplementation with high-oleic sunflower oil (HOSO) and/or α-tocopheryl acetate (VE), as well as the influence of storage conditions. Four different diets (control; HOSO; VE; HOSO+VE), were fed to swines until slaughtering. Meat slices were packed in vessels with transparent shrink film and exposed to white fluorescent light for 3 days at 8 °C. HOSO supplementation increased oleic acid content of pork meat. The highest levels of peroxide value (PV) and cholesterol oxidation products (COPs) were detected in the control group, whereas HOSO-enriched diets displayed the highest thiobarbituric reactive substance (TBARs) content. After storage under light exposure, pork meat slices exhibited a decrease of PV, which resulted in an increasing trend of TBARs and COPs. Feeding enrichment with both HOSO and vitamin E can be, therefore, used as an appropriate supplementation strategy to produce pork meat with a suitable oxidative stability.