Alleviating Heat Stress in Fattening Pigs: Low-Intensity Showers in Critical Hours Alter Body External Temperature, Feeding Pattern, Carcass Composition, and Meat Quality Characteristics.

Heat stress is a significant environmental problem that has a detrimental impact on animal welfare and production efficiency in swine farms. The current study was conducted to assess the effect of low-intensity showers, provided during critical high-temperature hours daily, on body external temperature, feeding pattern, and carcass and meat quality characteristics in fattening pigs. A total of 400 animals (200 barrows and 200 gilts) were randomly allotted in 40 pens. A shower nozzle was installed over 20 pens (half barrows and half gilts) where pigs received a low-intensity shower for 2 min in 30 min intervals from 12 to 19 h (SHO group). Another group without showers was also considered (CON). Feeder occupancy measurement, thermographic measures, and carcass and meat quality parameters were studied. In the periods with higher environmental temperatures, SHO animals showed an increase in the feeder occupancy rate compared to the CON group. A decrease in temperature was observed after the shower, regardless of the anatomical location (p < 0.005). The treatment with showers led to higher values than in the CON group of 4.72%, 3.87%, 11.8%, and 15.1% for hot carcass weight, lean meat yield, and fat thickness in Longissimus Dorsi (LD) and Gluteus Medius muscles, respectively (p < 0.01). Pork from CON showed a 14.9% higher value of drip loss, and 18.9% higher malondialdehyde concentration than SHO (p < 0.01); meanwhile, intramuscular fat content was 22.8% higher in SHO than in CON (p < 0.01). On the other hand, the CON group exhibited higher L* (2.13%) and lower a* and b* values (15.8% and 8.97%) compared to the SHO group. However, the pH20h of the CON group was significantly lower than that of the SHO group (p < 0.001), indicating a softer pH decrease. Related to fatty acids in subcutaneous outer and inner layers and intramuscular fat, the CON group showed higher ΣSFA and lower ΣMUFA and Δ9-desaturase indexes than SHO (p < 0.05). In conclusion, the amelioration of heat stress through showers at critical times should be considered an interesting tool that improves both carcass and meat quality, as well as animal welfare.

Immunochemical Recognition of Bothrops rhombeatus Venom by Two Polyvalent Antivenoms.

The protein profile of Bothrops rhombeatus venom was compared to Bothrops asper and Bothrops atrox, and the effectiveness of antivenoms from the National Institute of Health of Colombia (INS) and Antivipmyn-Tri (AVP-T) of Mexico were analyzed. Protein profiles were studied with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and reverse-phase high-performance liquid chromatography (RP-HPLC). The neutralizing potency and the level of immunochemical recognition of the antivenoms to the venoms were determined using Western blot, affinity chromatography, and enzyme-linked immunosorbent assay (ELISA). Bands of phospholipase A2 (PLA2), metalloproteinases (svMPs) I, II, and III as well as serine proteinases (SPs) in the venom of B. rhombeatus were recognized by SDS-PAGE. With Western blot, both antivenoms showed immunochemical recognition towards PLA2 and svMP. INS showed 94% binding to B. rhombeatus venom and 92% to B. asper while AVP-T showed 90.4% binding to B. rhombeatus venom and 96.6% to B. asper. Both antivenoms showed binding to PLA2 and svMP, with greater specificity of AVP-T towards B. rhombeatus. Antivenom neutralizing capacity was calculated by species and mL of antivenom, finding the following for INS: B. asper 6.6 mgV/mL, B. atrox 5.5 mgV/mL, and B. rhombeatus 1.3 mgV/mL. Meanwhile, for AVP-T, the following neutralizing capacities were found: B. asper 2.7 mgV/mL, B. atrox 2.1 mgV/mL, and B. rhombeatus 1.4 mgV/mL. These results show that both antivenoms presented similarity between calculated neutralizing capacities in our trial, reported in a product summary for the public for the B. asper species; however, this does not apply to the other species tested in this trial.

Short- and Long-Term Effects of Split-Suckling in Pigs According to Birth Weight.

Forty-eight litters were used, with a total number of 645 piglets involved in the study. The split-suckling technique was applied to half of the litters at the end of farrowing by removing the heaviest piglets over three periods of 1 h. The piglets were individually weighed at 0, 1 d, and at weaning. Piglet losses were recorded daily. Traceability was maintained until the carcass splitting and meat analysis took place. Carcasses were eviscerated and weighed individually. Total mortality at weaning was affected by body weight, where the low-body-weight piglets showed a mortality rate almost four times higher than that of the normal-weight piglets. Mortality was highest in the first days of life, especially in the low-body-weight piglets. At weaning, split-suckling treatment caused a slight increase in mortality compared to the control group piglets (25% vs. 17.1%). Split-suckling had a positive effect on weight gain during the first 24 h of life (p = 0.014), and there was an interaction between treatment and parity (p = 0.007), with split-suckling being more effective in the primiparous sows compared to the multiparous sows. The piglets from litters receiving the split-suckling treatment had a lower average daily gain during the lactation period (p < 0.001) than the piglets from the control group. Weight gain during the first 24 h of life of the piglets subjected to split-suckling was higher than those of the control group. A lower IgG and α-tocopherol in plasma in the heavier piglets subjected to split-suckling treatment was observed in comparison to their respective control. The piglets from litters receiving the treatment showed a lower average daily gain during the lactation period (p < 0.001) than the piglets from the control group. No difference in slaughter weight was observed according to treatment. The pigs which received split-suckling treatment showed lower subcutaneous fat thickness (p < 0.0013) and higher lean meat yield (p < 0.0027), this effect being more marked in pigs from primiparous sows. Intramuscular fat concentration was higher in the Longissimus Dorsi muscle of the low-body-weight piglets. In the pigs that received split-suckling treatment, a higher concentration of C18:3n-3 (p = 0.036) and a tendency towards a higher concentration of C18:2n-6 (p = 0.107) and unsaturation index (p = 0.113) was observed in intramuscular fatty acids at slaughter, together with a lower concentration of C16:0 (p = 0.053) and SFA (p = 0.064). In conclusion, long-term response to split-suckling, particularly in low-birth-weight piglets, suggests an alteration in adiposity and metabolic regulation in these piglets that receive high levels of colostrum.

Short- and Long-Term Effects of Birth Weight and Neonatal Care in Pigs.

Swine industries worldwide face a loss in profit due to high piglet mortality, particularly as a consequence of the marked increase in prolificity and low birth weight (BW) of piglets. This research studied the effect of BW and individual neonatal care provided to piglets on preweaning mortality, and the long-term effects on growth and carcass and meat characteristics. Litters from seventy-one crossbred sows (PIC 34) were included in the trial. Half of each litter did not receive any further management, and the remaining half received the pre-established management protocol of early assistance of neonatal care (NC). Along lactation, the low-BW piglets (weight equal to or less than 1.1 kg) showed a threefold higher mortality rate than piglets of higher weights (32 vs. 10%; p = 0.001), with mortality particularly concentrated within the first week after birth. No effect of NC treatment was observed on mortality ratio caused by crushing, but a significant effect was observed in low-BW piglets who died of starvation (p < 0.01). The effect of NC on growth is dependent on BW, and heavier piglets at birth benefit from NC treatment to a higher extent than low-BW piglets. Low-BW piglets showed a higher fatness (p = 0.003), lower lean cut yield (p = 0.002) in carcasses, and higher intramuscular fat (IMF) content (2.29% vs. 1.91%; p = 0.01) in meat. NC treatment increased the lean content in carcasses from low-BW piglets (p < 0.01). The monounsaturated fatty acids concentration was higher in lower-than-normal-BW piglets (48.1% vs. 47.1%; p = 0.002) and the opposite effect was observed for polyunsaturated fatty acids (13.6% vs. 15.7%; p = 0.002). NC treatment induced a higher concentration of n-7 fatty acids. In conclusion, NC treatment may be a useful practice to reduce mortality in low-BW piglets. Moreover, NC could affect carcass fatness and meat quality, thus suggesting a long-term effect on metabolism.

Free-Range Feeding Alters Fatty Acid Composition at the sn-2 Position of Triglycerides and Subcutaneous Fat Physicochemical Properties in Heavy Pigs.

The nutritional value of fat consumption depends on both the fatty acid composition and the positional distribution of fatty acids within the triglyceride molecule. This research studies the effect of feeding with three different diets (4% lard-enriched; 11.5% high-oleic sunflower-enriched; and extensive feeding mainly with acorns) on the composition of fatty acids in the sn-2 position (and sn-1,3) of triglycerides and the textural properties of subcutaneous fat in heavy Iberian pigs (n = 210 castrated males). A moderate dietary enrichment with oleic acid in mixed diets did not alter the regulation of the sn-2 position of triglyceride (69.9% and 13.9% of palmitic and oleic acids, respectively), but the extremely high intake of oleic acid in pigs fed mainly on acorns changed the proportions of palmitic and oleic acids at the sn-2 position in the subcutaneous fat of pigs (55.0% and 27.2%, respectively). Hardness, adhesiveness, cohesiveness, gumminess, and chewiness showed the least values in EXT pigs, and the greatest values in LARD-fed barrows. SUN cohesiveness and gumminess did not differ from those fed LARD. In addition, Iberian pigs raised in free-range conditions had a more favorable nutritional lipid profile for human health compared to pigs fed conventional diets.

Proanthocyanidins Maintain Cardiac Ionic Homeostasis in Aldosterone-Induced Hypertension and Heart Failure.

Excess aldosterone promotes pathological remodeling of the heart and imbalance in cardiac ion homeostasis of sodium, potassium and calcium. Novel treatment with proanthocyanidins in aldosterone-treated rats has resulted in downregulation of cardiac SGK1, the main genomic aldosterone-induced intracellular mediator of ion handling. It therefore follows that proanthocyanidins could be modulating cardiac ion homeostasis in aldosterone-treated rats. Male Wistar rats received aldosterone (1 mg kg-1 day-1) +1% NaCl for three weeks. Half of the animals in each group were simultaneously treated with the proanthocyanidins-rich extract (80% w/w) (PRO80, 5 mg kg-1 day-1). PRO80 prevented cardiac hypertrophy and decreased calcium content. Expression of ion channels (ROMK, NHE1, NKA and NCX1) and calcium transient mediators (CAV1.2, pCaMKII and oxCaMKII) were reduced by PRO80 treatment in aldosterone-treated rats. To conclude, our data indicate that PRO80 may offer an alternative treatment to conventional MR-blockade in the prevention of aldosterone-induced cardiac pathology.

Effects of Low Phytanic Acid-Concentrated DHA on Activated Microglial Cells: Comparison with a Standard Phytanic Acid-Concentrated DHA.

Docosahexaenoic acid (DHA, 22:6 n-3) is an essential omega-3 (ω-3) long chain polyunsaturated fatty acid of neuronal membranes involved in normal growth, development, and function. DHA has been proposed to reduce deleterious effects in neurodegenerative processes. Even though, some inconsistencies in findings from clinical and pre-clinical studies with DHA could be attributed to the presence of phytanic acid (PhA) in standard DHA treatments. Thus, the aim of our study was to analyze and compare the effects of a low PhA-concentrated DHA with a standard PhA-concentrated DHA under different neurotoxic conditions in BV-2 activated microglial cells. To this end, mouse microglial BV-2 cells were stimulated with either lipopolysaccharide (LPS) or hydrogen peroxide (H2O2) and co-incubated with DHA 50 ppm of PhA (DHA (PhA:50)) or DHA 500 ppm of PhA (DHA (PhA:500)). Cell viability, superoxide anion (O2-) production, Interleukin 6 (L-6), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), glutathione peroxidase (GtPx), glutathione reductase (GtRd), Caspase-3, and the brain-derived neurotrophic factor (BDNF) protein expression were explored. Low PhA-concentrated DHA protected against LPS or H2O2-induced cell viability reduction in BV-2 activated cells and O2- production reduction compared to DHA (PhA:500). Low PhA-concentrated DHA also decreased COX-2, IL-6, iNOS, GtPx, GtRd, and SOD-1 protein expression when compared to DHA (PhA:500). Furthermore, low PhA-concentrated DHA increased BDNF protein expression in comparison to DHA (PhA:500). The study provides data supporting the beneficial effect of low PhA-concentrated DHA in neurotoxic injury when compared to a standard PhA-concentrated DHA in activated microglia.

Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry.

There is increasing evidence suggesting the important role of inflammation and, subsequently, macrophages in the development and progression of renal disease. Macrophages are heterogeneous cells that have been implicated in kidney injury. Macrophages may be classified into two different phenotypes: classically activated macrophages (M1 macrophages), that release pro-inflammatory cytokines and promote fibrosis; and alternatively activated macrophages (M2 macrophages) that are associated with immunoregulatory and tissue-remodeling functions. These macrophage phenotypes need to be discriminated and analyzed to determine their contribution to renal injury. However, there are scarce studies reporting consistent phenotypic and functional information about macrophage subtypes in inflammatory renal disease models, especially in rats. This fact may be related to the limited macrophage markers used in rats, contrary to mice. Therefore, novel strategies are necessary to quantify and characterize the renal content of these infiltrating cells in a reliable way. This manuscript details a protocol for kidney digestion and further phenotypic and quantitative analysis of macrophages from rat kidneys by flow cytometry. Briefly, kidneys were incubated with collagenase and total macrophages were identified according to the dual presence of CD45 (leukocytes common antigen) and CD68 (PAN macrophage marker) in live cells.This was followed by surface staining of CD86 (M1 marker) and CD163 (M2 marker). Rat peritoneal macrophages were used as positive control for macrophage marker detection by flow cytometry. Our protocol resulted in low cellular mortality and allowed characterization of different intracellular and surface protein markers, thus limiting the loss of cellular integrity observed in other protocols. Moreover, this procedure allows the use of macrophages for further techniques, including cell sorting and mRNA or protein expression studies, among others.

Proanthocyanidins block aldosterone-dependent up-regulation of cardiac gamma ENaC and Nedd4-2 inactivation via SGK1.

Aldosterone plays a central role in the development of cardiac pathological states involving ion transport imbalances, especially sodium transport. We have previously demonstrated a cardioprotective effect of proanthocyanidins in aldosterone-treated rats. Our objective was to investigate for the first time the effect of proanthocyanidins on serum and glucocorticoid-regulated kinase 1 (SGK1), epithelial Na+ channel (γ-ENaC), neuronal precursor cells expressed developmentally down-regulated 4-2 (Nedd4-2) and phosphoNedd4-2 protein expression in the hearts of aldosterone-treated rats. Male Wistar rats received aldosterone (1mg kg-1day-1)+1% NaCl for 3weeks. Half of the animals in each group were simultaneously treated with the proanthocyanidins-rich extract (80% w/w) (PRO80, 5mg kg-1day-1). Hypertension and diastolic dysfunction induced by aldosterone were abolished by treatment with PRO80. Expression of fibrotic, inflammatory and oxidative mediators were increased by aldosterone-salt administration and blunted by PRO80. Antioxidant capacity was improved by PRO80. The up-regulated aldosterone mediator SGK1, ENaC and p-Nedd4-2/total Nedd4-2 ratio were blocked by PRO80. PRO80 blunted aldosterone-mineralocorticoid-mediated up-regulation of ENaC provides new mechanistic insight of the beneficial effect of proanthocyanidins preventing the cardiac alterations induced by aldosterone excess.

Aldosterone Induces Renal Fibrosis and Inflammatory M1-Macrophage Subtype via Mineralocorticoid Receptor in Rats.

We aimed to evaluate macrophages heterogeneity and structural, functional and inflammatory alterations in rat kidney by aldosterone + salt administration. The effects of treatment with spironolactone on above parameters were also analyzed. Male Wistar rats received aldosterone (1 mgkg-1d-1) + 1% NaCl for 3 weeks. Half of the animals were treated with spironolactone (200 mg kg-1d-1). Systolic and diastolic blood pressures were elevated (p<0.05) in aldosterone + salt-treated rats. Relative kidney weight, collagen content, fibronectin, macrophage infiltrate, CTGF, Col I, MMP2, TNF-α, CD68, Arg2, and SGK-1 were increased (p<0.05) in aldosterone + salt-treated rats, being reduced by spironolactone (p<0.05). Increased iNOS and IFN-γ mRNA gene expression (M1 macrophage markers) was observed in aldosterone + salt rats, whereas no significant differences were observed in IL-10 and gene ArgI mRNA expression or ED2 protein content (M2 macrophage markers). All the observed changes were blocked with spironolactone treatment. Macrophage depletion with liposomal clodronate reduced macrophage influx and inflammatory M1 markers (INF-γ or iNOS), whereas interstitial fibrosis was only partially reduced after this intervention, in aldosterone plus salt-treated rats. In conclusion, aldosterone + salt administration mediates inflammatory M1 macrophage phenotype and increased fibrosis throughout mineralocorticoid receptors activation.

Lower Oral Doses of Micellized α-Tocopherol Compared to α-Tocopheryl Acetate in Feed Modify Fatty Acid Profiles and Improve Oxidative Status in Pigs.

This study evaluated the effect of vitamin E supplementation source, and the dose given to sows or piglets, on the fatty acid profile of colostrum, milk, subcutaneous and intramuscular fat, and the oxidative status of piglets at 39 days of age. Sows (n = 10) were given 150 mg dl-α-tocopheryl acetate/d in feed, or 75 or 50 mg micellized-d-α-tocopherol/d in water from Day 103 of pregnancy. Weaning piglets from each group of sows (n = 7) received 3.33 mg dl-α-tocopheryl acetate/d in feed, or 1.7 mg micellized-d-α-tocopherol/d or 1.1 mg micellized-d-α-tocopherol/d in water for 14 days. Colostrum from sows supplemented with micellized-d-α-tocopherol had a lower proportion of C20:0 (P = 0.02), C18:4 n-3 (P = 0.03) and a higher C18:1 n-9 to C18:0 ratio than those given dl-α-tocopheryl acetate. Supplementation with micellized-d-α-tocopherol decreased the C18:0 proportion (P = 0.04) and the C18:1 n-9 to C18:0 ratio (P = 0.03) in milk, whereas the C18:1 n-7 proportion increased (P = 0.03) compared to dl-α-tocopheryl acetate. Composition was affected by the d-α-tocopherol dose. A similar trend to that observed in milk was observed in fatty acid composition in piglet fat. Piglets supplemented with micellized-d-α-tocopherol at low doses did not have different ferric reducing antioxidant power in muscle tissues (P = 0.31) than when they were supplemented with dl-α-tocopheryl acetate. Piglets given 1.7 mg micellized-d-α-tocopherol/d had lower oxidized glutathione than those given 1.1 mg/d (P = 0.0055). In conclusion, oral supplementation of sows (75 mg/d) and piglets (1.7 mg/d) with micellized natural vitamin E modified the fatty acid profile of piglet tissues and improved their oxidative status.

Effect of age at the beginning of the free-range fattening period on growth and carcass and fat quality in Iberian pigs.

This experiment was carried out to study the influence of age at the beginning of the free-range fattening period (traditional pigs, TP, age 12 months vs. young pigs, YP, age 8 months) on the performance of Iberian pigs. During 152 days prior to the fattening period, TP and YP pigs received 1.7 and 2.6 kg feed per day, respectively. During fattening, TP pigs had a higher average daily gain (p < 0.05) than YP pigs. The proportions of PUFA and n-3 fatty acids of the outer and inner layers of subcutaneous backfat were higher in TP than in YP pigs (p < 0.05), while the proportions of C16:0 and SFA in the inner layer of subcutaneous backfat were greater in YP than in TP pigs (p < 0.05). The ratio of n-6/n-3 in subcutaneous backfat was lower in TP than in YP pigs (p < 0.05). The percentage of intramuscular fat in longissimus dorsi muscle was higher in TP than in YP pigs (p < 0.05). The relationship between the percentage of intramuscular fat in longissimus dorsi muscle and average daily gain during the free-range fattening period adjusted to a quadratic function (p < 0.05). The concentration of alpha- and gamma-tocopherol in subcutaneous backfat at slaughter was significantly higher in TP than in YP pigs (p < 0.05). It is concluded that Iberian pigs that have 8 months of age at the beginning of free-range feeding have adequate commercial quality.