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Objective: Coronary artery disease remains a leading cause of morbidity and mortality worldwide. Patients with advanced coronary artery disease who are not eligible for endovascular or surgical revascularization have limited options. Extracellular vesicles have shown potential to improve myocardial function in preclinical models. Extracellular vesicles can be conditioned to modify their components. Hypoxia-conditioned extracellular vesicles have demonstrated the ability to reduce infarct size and apoptosis in small animals. Our objective is to assess the potential benefits of hypoxia-conditioned extracellular vesicles in a large animal model of coronary artery disease. Methods: Coronary artery disease was induced in 14 Yorkshire swine by ameroid constriction of the left circumflex coronary artery. Two weeks postsurgery, swine underwent a repeat left thoracotomy for injections of hypoxia-conditioned extracellular vesicles (n = 7) or saline (control, n = 7). Five weeks later, all animals underwent terminal harvest for perfusion measurements and myocardial sectioning. Results: Myocardial perfusion analysis demonstrated a trend toward increase at rest and a significant increase during rapid pacing (P = .09, P

Investigations on acute carbon monoxide (CO) poisoning struggle to highlight a relevant discriminant criterion related to CO poisoning severity for predicting complications, such as delayed neurological syndromes. In this context, it remains difficult to demonstrate the superiority of one method of oxygen (O2) administration over others or to identify the optimal duration of normobaric 100% oxygen (NBO) treatment. Myoglobin, as hemoglobin, are a potential binding site for CO, which could be a source of extravascular CO storage that impacts the severity of CO poisoning. It is not possible in routine clinical practice to estimate this potential extravascular CO storage. Indirect means of doing so that are available in the first few hours of poisoning could include, for example, the carboxyhemoglobin half-life (COHbt1/2), which seems to be influenced itself by the level and duration of CO exposure affecting this store of CO within the body. However, before the elimination of CO can be assessed, the COHbt1/2 toxicokinetic model must be confirmed: research still debates whether this model mono- or bi-compartmental. The second indirect mean could be the assessment of a potential COHb rebound after COHb has returned to 5% and NBO treatment has stopped. Moreover, a COHb rebound could be considered to justify the duration of NBO treatment. On an experimental swine model exposed to moderate CO poisoning (940 ppm for ±118 min until COHb reached 30%), we first confirm that the COHb half-life follows a bi-compartmental model. Secondly, we observe for the first time a slight COHb rebound when COHb returns to 5% and oxygen therapy is stopped. On the basis of these two toxicokinetic characteristics in favor of extravascular CO storage, we recommend that COHbt1/2 is considered using the bi-compartmental model in future clinical studies that compare treatment effectiveness as a potential severity criterion to homogenize cohorts of the same severity. Moreover, from a general toxicokinetic point of view, we confirm that a treatment lasting less than 6 hours appears to be insufficient for treating moderate CO poisoning.

Background: Induced hypothermia post-cardiac arrest is neuroprotective in animal experiments, but few high-quality studies have been performed in larger animals with human-like brains. The neuroprotective effect of postischemic hypothermia has recently been questioned in human trials. Our aim is to investigate whether hypothermia post-cardiac arrest confers a benefit compared to normothermia in large adult animals. Our hypothesis is that induced hypothermia post cardiac arrest is neuroprotective and that the effect diminishes when delayed two hours. Methods: Adult female pigs were anesthetized, mechanically ventilated and kept at baseline parameters including normothermia (38 °C). All animals were subjected to ten minutes of cardiac arrest (no-flow) by induced ventricular fibrillation, followed by four minutes of cardiopulmonary resuscitation with mechanical compressions, prior to the first countershock. Animals with sustained return of spontaneous circulation (systolic blood pressure >60 mmHg for ten minutes) within fifteen minutes from start of life support were included and randomized to three groups; immediate or delayed (2 h) intravenous cooling, both targeting 33 °C, or intravenously controlled normothermia (38 °C). Temperature control was applied for thirty hours including cooling time, temperature at target and controlled rewarming (0.5 °C/h). Animals were extubated and kept alive for seven days. The primary outcome measure is histological brain injury on day seven. Secondary outcomes include neurological and neurocognitive recovery, and the trajectory of biomarkers of brain injury. Conclusion: High-quality animal experiments in clinically relevant large animal models are necessary to close the gap of knowledge regarding neuroprotective effects of induced hypothermia after cardiac arrest.Trial registration: Preclinicaltrials.eu (PCTE0000272), published 2021-11-03.

In vivo delivery of mRNA is promising for the study of gene expression and the treatment of diseases. Lipid nanoparticles (LNPs) enable efficient delivery of mRNA constructs, but protein expression has been assumed to be limited to the liver. With specialized LNPs, delivery to extrahepatic tissue occurs in small animal models; however, it is unclear if global delivery of mRNA to all major organs is possible in humans because delivery may be affected by differences in innate immune response and relative organ size. Furthermore, limited studies with LNPs have been performed in large animal models, such as swine, due to their sensitivity to complement activation-related pseudoallergy (CARPA). In this study, we found that exogenous protein expression occurred in all major organs when swine were injected intravenously with a relatively low dose of mRNA encapsulated in a clinically relevant LNP formulation. Exogenous protein was detected in the liver, spleen, lung, heart, uterus, colon, stomach, kidney, small intestine, and brain of the swine without inducing CARPA. Furthermore, protein expression was detected in the bone marrow, including megakaryocytes, hematopoietic stem cells, and granulocytes, and in circulating white blood cells and platelets. These results show that nearly all major organs contain exogenous protein expression and are viable targets for mRNA therapies.

Heat stress (HS) negatively impacts a variety of production parameters in growing pigs; however, the impact of biological sex on the HS response is largely unknown. To address this, 48 crossbred barrows and gilts (36.8 ± 3.7 kg BW) were individually housed and assigned to one of three constant environmental conditions: (1) thermoneutral (TN) (20.8 ± 1.6 °C; 62.0 ± 4.7% relative humidity; n = 8/sex), (2) HS (39.4 ± 0.6 °C; 33.7 ± 6.3% relative humidity) for 1 d (HS1; n = 8/sex), or (3) or for 7 d (HS7; n = 8/sex). As expected, HS increased rectal temperature (Tr) following 1 d of HS (1.0 °C; P

Introduction: Resuscitative endovascular balloon occlusion of the aorta (REBOA) has emerged as an adjunct to CPR for nontraumatic cardiac arrest (NTCA). This translational study investigated the impact of varying low-flow duration (15- vs 30-mins) on REBOA's hemodynamic performance and ability to achieve return of spontaneous circulation (ROSC) in a porcine model. Methods: Thirty-two pigs were anesthetized and placed into ventricular fibrillation. All animals received a 4-min no-flow period before CPR was initiated. Animals were randomized into four groups: 15- vs 30-minutes of CPR; REBOA vs. no-REBOA. After completion of 15- or 30-minute low-flow, ACLS was initiated and REBOA was inflated in experimental animals. Results: In the 15-mins groups, there were no differences in the rates of ROSC between REBOA (4/8, 50%) and control (4/8, 50%; p = 0.99). However, in the 30-min groups, the REBOA animals had a significantly higher rate of ROSC (6/8, 75%) compared to control (1/8, 12.5%; p = 0.04). In the 7-mins after REBOA deployment in the 30-min animals there was a statistically significant difference in coronary perfusion pressure (REBOA 42.1 mmHg, control 3.6 mmHg, p = 0.038). Importantly, 5/6 animals that obtained ROSC in the 30-min group with REBOA re-arrested at least once, with 3/6 maintaining ROSC until study completion. Conclusion: In our porcine model of NTCA, REBOA preferentially improved hemodynamics and ROSC after a 30-mins period of low-flow CPR. REBOA may be a viable strategy to improve ROSC after prolonged downtime, however, more hemodynamic support will be required to maintain ROSC.

Paslahepevirus balayani (formerly known as hepatitis E virus) is an emerging cause of foodborne disease in Europe, transmitted mainly by the consumption of raw or undercooked pork. Since little is known about the presence of the virus in several pork products that are eaten uncooked, our aim was to evaluate the prevalence of Paslahepevirus balayani in groups of commercial pork products intended for human consumption subjected to different processing techniques. A total of 1265 samples of pork products from Spain were divided into four groups and tested for the presence of Paslahepevirus balayani RNA: unprocessed pig and wild boar meat frozen at −20 °C (n = 389), dry-cured pork products (n = 391), dry-cured and salted pork products (n = 219), and boiled products (n = 266) (none of these products contained pork liver). Five samples were positive for Paslahepevirus balayani RNA (overall prevalence: 0.4%; 95% CI: 0.17% - 0.92%). All positive samples were from unprocessed meat stored at −20 °C, with a prevalence in this group of 1.3% (95% CI: 0.42–3.44); two samples came from pigs (1.1%; 95% CI: 0.13–3.81) and three from wild boar (1.5%; 95% CI: 0.31–4.28). None of the pork samples in the other groups was positive. In conclusion, Paslahepevirus balayani was found in unprocessed swine products form Spain, but not in processed products intended to be consumed undercooked, demonstrating that transmission of this zoonotic virus by eating these pork products should be more seriously considered.

Background: Coronary artery disease remains a leading cause of death worldwide. Bone mesenchymal stem cell–derived extracellular vesicles (EVs) have shown promise in the setting of myocardial ischemia. Furthermore, the properties of the EVs can be modified via preconditioning of progenitor cells. Previous research from our lab demonstrated a significant decrease in proinflammatory signaling following treatment with EVs derived from starvation preconditioning of human bone mesenchymal stem cells (MVM EVs) in a porcine model of chronic myocardial ischemia. However, rodent models have demonstrated that the use of EVs derived from hypoxia preconditioning of bone mesenchymal stem cells (HYP EVs) may have extended benefits compared to MVM EVs. This study evaluated the effect of HYP EVs on inflammation in a swine model of chronic myocardial ischemia. We hypothesized that HYP EVs would have a greater anti-inflammatory effect than MVM EVs or saline (CON). Methods: Yorkshire swine fed a standard diet underwent placement of an ameroid constrictor to the left circumflex artery. Two weeks later, the animals received intramyocardial injection of saline (CON; n = 6), starvation-derived EVs (MVM; n = 10), or hypoxia-derived EVs (HYP; n = 7). After 5 weeks, myocardial perfusion was assessed, and left ventricular myocardial tissue was harvested. Protein expression was measured using immunoblotting. Data were analyzed via the Kruskal-Wallis test or one-way analysis of variance based on the results of a Shapiro-Wilk test. Coronary perfusion was plotted against relative cytokine concentration and analyzed with the Spearman rank-sum test. Results: HYP EV treatment was associated with decreased expression of proinflammatory markers interleukin (IL)-6 (P = .03), Pro-IL-1ß (P = .01), IL-17 (P

Background: Preclinical animal models which mimic the dimensions of long urethral strictures (>2 cm in length) encountered in the clinic are necessary to evaluate prospective graft designs for urethroplasty. The purpose of this study was to develop both male and female porcine models of long urethral strictures (∼4 cm in length) and characterize histological and functional outcomes of iatrogenic stricture formation between genders. Methods: Focal, partial thickness urethral injuries were created over 5–6 cm long segments in male and female swine (N = 4 per gender) via electrocoagulation and the degree of stricture formation was monitored for up to 6 weeks by urethroscopy and retrograde urethrography. Animals were sacrificed following stricture confirmation and histological, immunohistochemical, and histomorphometric analyses were performed on strictured and uninjured control urethral segments to profile wound healing responses. Results: Urethral stricture formation was detected in all female swine by 2 weeks and 100 % of male swine at 3.2 ± 1.8 weeks, post-operatively. The mean length of urethral strictures in both male and female swine was ∼4 cm. Substantial variations in the degree of stricture severity between sexes were observed with males exhibiting significant urethral stenosis and loss of α-smooth muscle actin+ smooth muscle bundles in comparison to controls, while females primarily displayed defects in pan-cytokeratin+ epithelia as well as functional urethral obstruction. Conclusions: Electrocoagulation injury is sufficient to produce long urethral strictures in male and female swine and the degree of stricture severity and nature of urethral obstruction was observed to be dependent on gender. Animal Protocol: AUP-19-150. Key message: Novel male and female models of long urethral strictures in swine were created to characterize histological and functional outcomes of iatrogenic stricture formation between genders.

ABSTRACT: Objectives: The aim of this study was to characterize the floR-carrying plasmids originating from Glaesserella parasuis and Actinobacillus indolicus isolated from pigs with respiratory disease in China. Methods: A total of 125 G. parasuis and 28 A. indolicus strains collected between 2009 and 2022 were screened for florfenicol resistance. Characterization of floR-positive isolates and plasmids were determined by antimicrobial susceptibility testing, serotyping, multilocus sequence typing (MLST), conjugation and transformation assays, whole-genome sequencing (WGS), and phylogenetic analysis. Results: One A. indolicus and six G. parasuis were identified as positive for floR. The six G. parasuis were assigned to four different serovars, including serovars 6, 7, 9, and unknown. In addition to strain XP11, six floR genes were located on plasmids. The six floR-bearing plasmids could be transformed into Pasteurella multocida and divided into two different types, including ∼5000 bp and ∼6000 bp plasmids. The ∼5000 bp plasmids consisting of rep, lysR, mobB, and floR genes, exhibited high similarity among Pasteurellaceae bacteria. Furthermore, the ∼6000 bp plasmids, consisting of rep, lysR, mobC, mobA/L, and floR genes, showed high similarity between G. parasuis and Actinobacillus Spp. Notably, WGS results showed that the floR modules of the two types of plasmids could be transferred and integrated into the diverse Pasteurellaceae- origined plasmids. Conclusion: This study firstly reported the characterization of floR-carrying plasmids from A. indolicus and a non-virulent serovar of G. parasuis in pigs in China and elucidated the transmission mechanism of the floR resistance gene among the Pasteurellaceae family.

There is a gap in the understanding of the relationship between dietary phytate levels and the relative efficacy of phytase to improve amino acid (AA) digestibility in pigs and chickens. Two experiments were conducted to investigate the effect of exogenous phytase on standardized ileal digestibility (SID) of AA and the apparent ileal digestibility (AID) of P in both standard- (SP) and high-phytate (HP) diets for broilers and swine. There were either 40 cages of Cobb 500 male broilers or 10 crossbred barrows (35 kg) fitted with ileal T-cannulas. Both studies were allotted to five dietary treatments (8 replicates). Treatments consisted of four corn-soybean meal-based diets arranged in a 2 × 2 factorial of standard or high phytate and exogenous phytase at 0 or 1 000 phytase units (FYT)/kg; and one N-free diet. Birds were fed a common starter diet from d 0 to 20 and fed experimental diets from d 20 to 25. Birds were euthanized on d 25 via CO2 asphyxiation, and digesta were collected from the terminal ileum. Pigs were fed for a total of four 7-d periods, where digesta were collected on d 6 and 7 of each period. Diet and digesta samples were analyzed for DM, N, Ti, AA, and P to determine AA and P digestibility. The SID of AA was determined by correcting the AID of AA for the basal endogenous losses estimated using the N-free diet. Main effects of the diet type (standard or HP) and phytase (0 or 1 000 FYT/kg), and the interaction of diet type and phytase were evaluated. For both experiments, the HP diets produced lower SID of AA compared to the SP (P

Objective: Limited treatments exist for nonoperative chronic coronary artery disease. Previously, our laboratory has investigated extracellular vesicle (EV) therapy as a potential treatment for chronic coronary artery disease using a swine model and demonstrated improved cardiac function in swine treated with intramyocardial EV injection. Here, we seek to investigate the potential cardiac benefits of EVs by using hypoxia-conditioned EVs (HEV). Specifically, this study aims to investigate the effect of HEV on apoptosis in chronically ischemic myocardium in swine. Methods: Fourteen Yorkshire swine underwent placement of an ameroid constrictor on the left circumflex artery. Two weeks later, swine underwent redo left thoracotomy with injection of either saline (control, n = 7) or HEVs (n = 7). After 5 weeks, swine were euthanized for tissue collection. Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to quantify apoptosis. Immunoblotting was used for protein quantification. Results: Terminal deoxynucleotidyl transferase dUTP nick end labeling staining showed a decrease in apoptosis in the HEV group compared with the control (P = .049). The HEV group exhibited a significant increase in the anti-apoptotic signaling molecule phospho-BAD (P = .005), a significant decrease in B-cell lymphoma 2 (P = .006) and an increase in the phospho-B-cell lymphoma to B-cell lymphoma 2 ratio (P

Background: Sepsis and septic shock still represent great challenges in critical care medicine. Sildenafil has been largely used in the treatment of pulmonary arterial hypertension, but its effects in sepsis are unknown. The aim of this study was to investigate the hypothesis that sildenafil can attenuate endotoxin-induced pulmonary hypertension in a porcine model of endotoxemia. Methods: Twenty pigs were randomly assigned to Control group (n.ß=.ß10), which received saline solution; or to Sildenafil group (n.ß=.ß10), which received sildenafil orally (100.ßmg). After 30.ßminutes, both groups were submitted to endotoxemia with intravenous bacterial lipopolysaccharide endotoxin (LPS) infusion (4.ß..g.kg-1.h-1) for 180.ßminutes. We evaluated hemodynamic and oxygenation functions, and also lung histology and plasma cytokine (TNF.., IL-1.., IL6, and IL10) and troponin I response. Results: Significant hemodynamic alterations were observed after 30.ßminutes of LPS continuous infusion, mainly in pulmonary arterial pressure (from Baseline 19.ß...ß2.ßmmHg to LPS30 52.ß...ß4.ßmmHg, p.ß

Pig homogeneity and growth are major concerns for the pig industry today. Variability in pigs’ size has a strong impact on profitability as uniformity plays a key role in the overall economic value of pigs produced. This research focused on statistical methods to identify pigs at risk of growth retardation at different stages of production. Data from 125 083 Yorkshire pigs at weaning (18–28 d), 59 533 pigs at the end of the nursery period (70–82 d) and 48 862 pigs at slaughter (155–170 d) were analyzed under three different cut-points (lowest 10, 20 and 30%) to characterize light animals. Records were randomly split into 2:1 training:testing sets, and each training data set was analyzed through an ordinary least squares approach and four machine learning algorithms (decision tree, random forest, and two alternative boosting approaches). A wide range of weighting functions were applied to give increased relevance to lighter pigs. Each resulting classification norm was used to classify light pigs in the testing data set. Both sensitivity and specificity were retained to construct the receiver operating characteristic curve, and the statistical performance of each analytical approach was evaluated by the area under the curve (AUC). In all production stages and cut-points, the random forest machine learning algorithm provided the highest AUC, closely followed by boosting procedures. For weaning BW (WW), factors related to birth BW and litter size accounted for more than 75% of the important prediction factors for light pigs. BW at the end of the nursery period and slaughter BW analyses revealed a similar pattern where WW and BW at the end of the nursery period accounted for more than 40 and 50% of statistical importance among the prediction factors, respectively. Machine learning algorithms are useful tools to easily evaluate the risk factors affecting the efficiency and homogeneity in swine. Since the BW at birth and weaning are key factors, sow nutrition and feeding management during gestation and lactation, along with piglet management during lactation, are identified as important influences on pig weight variability.

Former foodstuff products (FFPs) are promising alternative ingredients for reducing the waste of natural resources and the environmental impact of food production. This study investigates the effects of salty and sugary FFPs on growth performance, apparent total tract digestibility (ATTD), and growing-finishing pigs' empty body and carcass composition. Thirty-six Swiss Large White male castrated pigs were assigned to three growing (G) and finishing (F) diets: (1) standard diet (ST), 0% FFPs; (2) 30% conventional ingredients replaced by sugary FFPs (SU); and (3) 30% conventional ingredients replaced by salty FFPs (SA). Faecal samples from 24 selected pigs were collected to assess the ATTD of gross energy, crude fibres, and CP. The BW was measured weekly, while feed intake was determined daily. Average daily gain (ADG), average daily feed intake (ADFI), feed conversion ratio (FCR), and ATTD were calculated for both the growing and finishing periods. Pigs' body composition was determined at ≥20 and ≥98 kg using dual-energy X-ray absorptiometry. In the growing but not finishing period, the FCR was lower (P 0.05) ADG, ADFI, FCR, or BW at slaughter. In both the growing and finishing periods, the gross energy ATTD was higher (P 0.05) effects on the parameters related to the pigs' body composition (e.g., average daily fat weight gain). Finally, the carcasses of the SU group had the thickest belly fat, even though the total fat content was similar among the groups. This study confirms that including FFPs has no detrimental effects on growth performance or live body/carcass composition in growing and finishing pigs.

The BIOPIGEE project (part of the One Health European Joint Programme under Horizon 2020) aimed to identify relevant measures to effectively control Salmonella, and another zoonotic pathogen, hepatitis E virus (HEV) within the pig meat food chain. The aim of this study was to identify biosecurity measures or management practices that are relevant for limiting Salmonella and/or HEV occurrence and spread within pig slaughterhouses. This was with the final goal of compiling a list of biosecurity measures for different processes and operations along the slaughter line with evidence of their effectiveness. To achieve this, a literature review was conducted on studies estimating the effectiveness of measures applied in slaughterhouses to reduce the microbial contamination of pig carcasses. Results of this literature search are discussed and presented in summary tables that could be used as a source of information for the pig slaughter industry to further develop their guidelines on hygienic slaughter.

Shortly after weaning, piglets generally eat dry feed poorly; but nevertheless, a phenomenal gain-to-feed ratio is achieved as they gain about as much weight as they eat (150–200 g/d). The high gain-to-feed ratio, though, cannot be explained by their nutrient intake or nutrient repartitioning. Analyses based on tissue composition and bio-electrical impedance data showed that newly weaned piglets lose fat, maintain protein, and gain large amounts of water because of edema. This edema, which may well contribute up to one kg of BW, seems to be triggered by refeeding syndrome. Refeeding syndrome in adult humans occurs when subjects fast for an extended period of time (weeks) that results in downshifts in metabolic activity and concomitant shedding of phosphate (PO4), magnesium (Mg), and potassium (K) in urine. If food is abruptly reintroduced, thus, resulting in strong insulin spikes, metabolism is triggered but hampered by a lack of PO4, Mg, K, and thiamine, causing hypophosphatemia, metabolic stress, and edema. In piglets, the same process appears to happen immediately after weaning but in hours rather than weeks, possibly linked to their high metabolic rate. Refeeding syndrome can be lethal in humans but does not appear to be directly lethal in piglets. Our attempts to prevent it through altered diet composition and/or controlled feeding programs have not resulted in better performance at the end of the nursery phase. A practical ramification of weaning-induced edema is that growth and gain-to-feed ratio data immediately after weaning should be interpreted with caution. In addition, diets arguably should be formulated to not strongly trigger insulin release, while high lysine levels are not needed as the gain is not based on protein accretion.

Swine nursery mortality is one of the most important key performance indicators (KPIs) for commercial swine production systems and, for the purpose of this study, refers to the number of pigs that died during the first 60 days during the post-weaning phase (approximately 170-day period). Likewise, nursery mortality accounts for almost 50% of the mortality that occurs throughout the post-weaning phase. Despite being a metric concerning the post-weaning phase of production, previous studies have demonstrated that nursery mortality is highly correlated with the previous performance of the piglets in the pre-weaning phase and to stocking conditions at placement in nursery sites. However, there is no evidence of that predictive models are currently routinely utilized in the swine industry to predict the nursery performance of individual groups of pigs based on their previous performance in the pre-weaning phase. One obstacle to building such predictive models is that health and management data as well as production data for the pre and post-weaning phases are collected with separate record-keeping programs and stored in unconnected databases.Thus, the objective of this study was to measure and compare the performance of 5 forecasting models for predicting nursery mortality using a master table containing data collected from a single swine production system for 1,831 groups of pigs. An automated model was built to integrate information for 48 variables included in the master table that was previously disconnected within the production system. After the model and parameter tuning process, the model with the best overall prediction performance was the Support Vector Machine (SVM) in terms of Root Mean Squared Error (RMSE=0.3929), Mean Absolute Error (MAE=0.2956), and coefficient of determination (R2=0.7761). This study demonstrated the ability to use an integrated master table containing whole-herd information until the placement event of nursery groups to predict their downstream nursery mortality through the initial 60 days of the post-weaning phase.

Changes in gastrointestinal architecture, high incidence of diarrhoea, and low feed intake (FI) are commonly observed around weaning of pigs, but the relationship between postweaning FI and diarrhoea is unclear. This study aimed to determine the effect of low or high FI during the first days after weaning on growth performance, diarrhoea probability, intestinal permeability, and morphology in pigs until postweaning day (PWD) 28. A total of 120 pigs (7.20 ± 0.26 kg) weaned at 28 days of age (PWD 0) were randomly allocated to five diets and housed individually until PWD 28. Two diets differed in CP and three diets differed in threonine and tryptophan levels. At PWD 4, pigs with the 25% lowest accumulated FI (LOW; n = 30) and 25% highest accumulated FI (HIGH; n = 30) were selected for the study. Faecal consistency was evaluated daily using a 4-scale visual scoring system. Blood was collected at PWD 4, 14, 21 and 28, and small intestinal and colonic tissue was obtained at slaughter on PWD 28. Until PWD 4, LOW pigs consumed approximately 20% (35.7 ± 5.9 g/day) of the FI of HIGH pigs (181 ± 5.75 g/day; P 0.10). The systemic inflammatory markers haptoglobin and C-reactive protein were higher for HIGH pigs at PWD 4 (P = 0.005), but not affected in the following periods (P > 0.10). Pigs in the HIGH group had an increased area of acidic mucin-producing cells in the small intestine compared with LOW pigs (P

Reducing the dietary CP concentration in the formulation of low-protein diets without adverse effects on animal growth performance and meat quality remains challenging. In this study, we investigated the effects of nicotinamide (NAM) on the nitrogen excretion, growth performance, and meat quality of growing-finishing pigs fed low-protein diets. To measure the nitrogen balance, we conducted two trials: in nitrogen balance trial 1, four crossbred (Duroc × Landrace × Large White) barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design with four diets and periods. The diets consisted of a basal diet + 30 mg/kg NAM (a control dose), basal diet + 90 mg/kg NAM, basal diet + 210 mg/kg NAM, and basal diet + 360 mg/kg NAM. In nitrogen balance trial 2, another four barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design. The diets consisted of a basal diet + including 30 mg/kg NAM (control), basal diet + 360 mg/kg NAM, low-protein diet + 30 mg/kg NAM, and low-protein diet + 360 mg/kg NAM. To measure growth performance, two trials were conducted. In growth performance trial 1, 40 barrows (37.0 ± 1.0 kg) were randomly allocated to one of four dietary treatments (n = 10 per group), whereas in growth performance trial 2, 300 barrows (41.4 ± 2.0 kg) were randomly allocated to one of four dietary treatments, with each dietary treatment conducted in five repetitions with 15 pigs each. The four diets in the two growth performance trials were similar to those in nitrogen balance trial 2. Supplementing the diet with 210 or 360 mg/kg NAM reduced urinary nitrogen excretion and total nitrogen excretion and increased nitrogen retention comparted with the control diet (P 0.05). Pigs fed the low-protein diet with 360 mg/kg NAM showed a decreased intramuscular fat content in the longissimus thoracis muscle when compared with pigs fed the control diet (P > 0.05). Our results suggest NAM as a suitable dietary additive to reduce dietary CP concentration, maximise nitrogen retention and growth performance, and decrease fat deposition in pigs.

The primary challenge of agriculture and livestock production is to face the growing competition between food, feed, fibre, and fuel, converting them from resource-intensive to resource-efficient. A circular economy approach, using agricultural by-products/co-products, in the livestock production system would allow to reduce, reuse, and redistribute the resources. Former food products (FFPs), also named ex-foods, could represent a valid option in strengthening resilience in animal nutrition. FFPs have a promising potential to be included regularly in animal diets due to their nutritive value, although their potential in animal nutrition remains understudied. A thorough investigation of the compositional and dietary features, thus, is essential to provide new and fundamental insights to effectively reuse FFPs as upgraded products for swine nutrition. Safety aspects, such as the microbial load or the presence of packaging remnants, should be considered with caution. Here, with a holistic approach, we review several aspects of FFPs and their use as feed ingredients: the nutritional and functional evaluation, the impact of the inclusion of FFPs in pigs’ diet on growth performance and welfare, and further aspects related to safety and sustainability of FFPs.

Pampa Rocha (PR) is a breed of pig that emerged in eastern Uruguay during the 18th century. They represent an important resource for non-intensive production using purebred or crossbred animals. However, productive activities have been oriented towards intensive production using commercial breeds, abandoning, except by some academic and educational institutions, the promotion of this creole breed. Thus, a population of few animals is still maintained, which could be in danger of disappearing. This work focuses on the fecal microbiota of these animals, which is related to the animal genetic background but also to their grazing capacity and resistance to weather. The structure and diversity of bacterial communities in the intestines of four PR adult females and of other breeds, including crosses, reared under non-grazing conditions, were analyzed and compared. Results obtained indicate that PR fecal microbiota is clearly different from those of other animals analyzed. Some sequences, corresponding to particular groups apparently related to the consumption of fiber, were strongly associated with PR pigs.

Pigs are considered to be the main reservoir for livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA), which is a zoonotic opportunistic pathogen. As LA-MRSA is an occupational hazard, there is an incentive to control its spread in pig herds. Currently, knowledge about effective control measures which do not require culling the whole herd are limited, and the control strategies against LA-MRSA vary between countries. This study uses a stochastic compartment model to simulate possible control measures for LA-MRSA in a farrow-to-finish pig herd. The aims of the study were to (1) extend a previously published disease spread model with additional management and control measures; (2) use the extended model to study the effect of the individual LA-MRSA control measures on the within-herd LA-MRSA prevalence; (3) evaluate the effect of control measures when they are implemented in combinations. From the individual control measures tested in the study, thorough cleaning was found to be most effective in reducing the LA-MRSA prevalence in the herd. When the different control measures were combined, cleaning together with disease surveillance had the largest impact on reducing the LA-MRSA and a higher chance of causing disease elimination. The results of the study showed that achieving disease elimination once LA-MRSA had been introduced in the herd was challenging but was more likely when control measures were introduced early during the outbreak. This emphasises the importance of early detection of the pathogen and subsequent rapid implementation of LA-MRSA control measures.

Giardia duodenalis is one of the common intestinal parasites causing diarrhea in humans and livestock, including pigs. Thus, a healthy livestock would result in a clean environment, which benefits humans. In the present study, the global molecular prevalence of G. duodenalis infection was determined in pig populations, through systematic exploration of 4 international databases (MEDLINE/PubMed, Scopus, Web of Science, and Google Scholar) until March 4th, 2022. A random-effects meta-analysis model was used to estimate the overall and subgroup-based pooled prevalence of G. duodenalis, and I2 index was used for the evaluation of the heterogeneity. Altogether, 42 datasets from 18 papers examined 7272 pigs across 12 nations, showing a 9.1% (95% CI: 5.6–14.3%) pooled molecular prevalence. Sensitivity analysis demonstrated no remarkable variation in the reported total prevalence upon removing individual studies. It was found that 6 Giardia assemblages (A-F) are capable to infect pigs around the world, including assemblage E [16 datasets, 41.1% (95% CI: 24.8–59.6%)], B [8 datasets, 28.2% (95% CI: 12.2–52.6%)], D [3 datasets, 16.2% (95% CI: 10.6–24.1%)], C [3 datasets, 11.6% (95% CI: 7.3–17.9%)], and A [11 datasets, 9.9% (95% CI: 5.6–16.9%)]. Of note, assemblage F was only reported in one study. Meta-regression analysis showed that publication year was not significantly associated with the Giardia prevalence in swine population, in contrast to the sample size. Substantially, animals in weaner and fattener stages were more prone to giardiasis. Assemblages A and B are of utmost zoonotic significance for humans, while assemblages C, D and F have, also, been found in dogs and cats. Still, little is known on the prevalence and distribution of Giardia assemblages in pigs and requires more extensive and detailed studies.

Litter traits are critical economic variables in the pig industry as they represent a production indicator that can serve to determine sow fertility. In this study, a genome-wide association study on litter traits, including total number born (TNB), number born alive (NBA), litter birth weight (LBW), average birth weight (ABW), and piglet uniformity (PU), was carried out on two pig breeds (Yorkshire and Landrace). A total of 3 637 pigs of both breeds were genotyped using the GeneSeek GGP Porcine 50K SNP BeadChip. A mixed linear model (MLM) and fixed and random model circulating probability unification (FarmCPU) were employed in the genome-wide association studies for litter traits using combined data from the two pig breeds and data from each breed separately. Additionally, the heritability of traits was estimated using three methods—pedigree-based best linear unbiased prediction (PBLUP), genomic best linear unbiased prediction (GBLUP), and single-step best linear unbiased prediction (ssGBLUP)—and was found to lie between 0.065 and 0.1289, 0.0478 and 0.0938, 0.0793 and 0.0935, 0.1862 and 0.2163, and 0.0327 and 0.0419 for TNB, NBA, LBW, ABW, and PU, respectively. We also compared the genomic prediction accuracies and unbiasedness for litter traits of the three BLUP models. Our results indicated that the ssGBLUP method provided higher predictive accuracies and more rational unbiasedness compared with the PBLUP and GBLUP methodologies. Furthermore, based on their possible roles, eight candidate genes (INHBA, LEPR, HDHD2, CTNND2, RNF216, HMX1, PAPPA2, and NTN1) were identified as being linked with litter traits. In the middle of the test, these genes were found to be connected with pig metabolism and ovulation rate. Our results provide the insights into the genetic architecture of litter traits in pigs, and the potential single nucleotide polymorphisms (SNPs) and candidate genes identified may benefit economic profits in pig-breeding industry and contribute to improve litter traits.

The spread of infectious livestock diseases is a major cause for concern in modern agricultural systems. In the dynamics of the transmission of such diseases, movements of livestock between herds play an important role. When constructing mathematical models used for activities such as forecasting epidemic development, evaluating mitigation strategies, or determining important targets for disease surveillance, including between-premises shipments is often a necessity. In the United States (U.S.), livestock shipment data is not routinely collected, and when it is, it is not readily available and mostly concerned with between-state shipments. To bridge this gap in knowledge and provide insight into the complete livestock shipment network structure, we have developed the U.S. Animal Movement Model (USAMM). Previously, USAMM has only existed for cattle shipments, but here we present a version for domestic swine. This new version of USAMM consists of a Bayesian model fit to premises demography, county-level livestock industry variables, and two limited data sets of between-state swine movements. The model scales up the data to simulate nation-wide networks of both within- and between-state shipments at the level of individual premises. Here we describe this shipment model in detail and subsequently explore its usefulness with a rudimentary predictive model of the prevalence of porcine epidemic diarrhea virus (PEDv) across the U.S. Additionally, in order to promote further research on livestock disease and other topics involving the movements of swine in the U.S., we also make 250 synthetic premises-level swine shipment networks with complete coverage of the entire conterminous U.S. freely available to the research community as a useful surrogate for the absent shipment data.

While biosecurity, a central component of the One Health concept, is clearly defined, a harmonized definition of the term ´biosecurity measure´ (BSM) is missing. In turn, particularly at the farm and policy level, this leads to misunderstandings, low acceptance, poor implementation, and thus suboptimal biosecurity along the food animal production chain. Moreover, different views on BSMs affects making comparisons both at the policy level as well as in the scientific community. Therefore, as part of the One Health EJP BIOPIGEE project, a work group i) collected and discussed relevant inclusion and exclusion criteria for measures to be considered in the context of biosecurity and ii) conducted a systematic literature review for potentially existing definitions for the term BSM. This exercise confirmed the lack of a definition of BSM, underlining the importance of the topic. In the pool of articles considered relevant to defining the term BSM, specific research themes were identified. Based on these outcomes, we propose a definition of the term BSM:“A biosecurity measure (BSM) – is the implementation of a segregation, hygiene, or management procedure (excluding medically effective feed additives and preventive/curative treatment of animals) that specifically aims at reducing the probability of the introduction, establishment, survival, or spread of any potential pathogen to, within, or from a farm, operation or geographical area.”The definition provides a basis for policymakers to identify factual BSMs, highlights the point of implementation and supports to achieve the necessary quality standards of biosecurity in food animal production. It also enables clear, harmonized, cross-sectoral communication of best biosecurity practices to and from relevant stakeholders and thus contribute to improving biosecurity and thereby strengthen the One Health approach.

Lowering crude protein in pig diets can reduce nitrogen (N) excretion and alter manure characteristics. Anaerobic digestion (AD) offers potential for converting pig manure into biogas and bio-based fertilizers (i.e., digestate). However, limited research exists on the effects of N content in pig manure on AD when pigs are fed diets with varying crude protein levels. This study investigated how lowering N content in pig manure through low crude protein diets may affect AD process stability, biogas generation, and digestate properties. Manures from different dietary treatments, named as control (CON), low N (LN), and very low N (VLN), with Total Kjeldahl Nitrogen concentrations of 5.87, 5.42, and 5.15 g/L, respectively, were investigated. Daily biogas production, composition (CH 4 , CO 2 , and H 2 S), and digestate properties were monitored over 13 fed-batch cycles (25 ± 4 days per cycle). The experiment was conducted at 20 ± 1 °C, a condition suited for milder climate regions, using six single-stage digesters operated in sequencing fed-batch mode. Data were analyzed by ANOVA using PROC MIXED with repeated measures. Results showed that the differences in N content in pig manure due to the three dietary treatments had a limited impact on biogas generation, with specific methane yields remaining similar over time. CH 4 concentrations remained stable between 60 and 65 %, ensuring high-quality biogas despite dietary variations. Differences between treatments became more pronounced with increased organic loading rates (OLRs) due to variations in the amount of volatile solids fed. AD also remained stable (Total Volatile Fatty Acids/Total Alkalinity <0.25) even at an OLR of 2.15 g of chemical oxygen demand L -1  day -1 , highlighting AD's robustness at lower temperatures. Digestate samples contained essential minerals beneficial for plant growth. More research is needed to explore varied manure compositions and feeding strategies to better understand the interactions of animal nutrition with AD., Competing Interests: Declaration of competing interest The authors declare that this research was conducted in the absence of any commercial or financial relationship that could be perceived as potential conflicts of interest., (Crown Copyright © 2024. Published by Elsevier B.V. All rights reserved.)

Dihydroquercetin (DHQ), also known as Taxifolin (TA), is a flavanonol with various biological activities, such as anticancer, anti-inflammatory, and antioxidative properties. It has been found to effectively increase the viability of porcine intestinal epithelial cells (IPEC-J2). However, the precise mechanism by which DHQ increases the proliferation of IPEC-J2 cells is not entirely understood. This study aimed to explore the potential pathways through which DHQ encourages the proliferation of IPEC-J2 cells. The findings indicated that DHQ significantly improved the protein expression of tight junction proteins (ZO-1, Occludin, and Claudin1) and a molecular biomarker of proliferation (PCNA) in IPEC-J2 cells. Furthermore, DHQ was found to increase the Wnt/β-catenin pathway-associated β-catenin, c-Myc, and cyclin D1 mRNA expression, and promote the protein expression of β-catenin and TCF4. To confirm the involvement of the Wnt/β-catenin signaling pathway in the DHQ-promoted proliferation of IPEC-J2 cells, the inhibitor LF3, which targets β-catenin/TCF4 interaction, was used. It was found that LF3 inhibited the protein expressions upregulated by DHQ and blocked the promotion of cell proliferation. These results indicate that DHQ positively regulates IPEC-J2 cell proliferation through the Wnt/β-catenin pathway, providing constructive insights into the role of DHQ in regulating intestine development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Antibiotic resistance (AMR) poses a significant global health challenge, with swine farms recognized as major reservoirs of antibiotic resistance genes (ARGs). Recently, bacterial membrane vesicles (BMVs) have emerged as novel carriers mediating horizontal gene transfer. However, little is known about the ARGs carried by BMVs in swine farm environments and their transfer potential. This study investigated the distribution, sources, and microbiological origins of BMVs in three key microbial habitats of swine farms (feces, soil, and fecal wastewater), along with the ARGs and mobile genetic elements (MGEs) they harbor. Characterization of BMVs revealed particle sizes ranging from 20 to 500 nm and concentrations from 10 8 to 10 12 particles/g, containing DNA and proteins. Metagenomic sequencing identified BMVs predominantly composed of members of the Proteobacteria phyla, including Pseudomonadaceae, Moraxellaceae, and Enterobacteriaceae, carrying diverse functional genes encompassing resistance to 14 common antibiotics and 74,340 virulence genes. Notably, multidrug resistance, tetracycline, and chloramphenicol resistance genes were particularly abundant. Furthermore, BMVs harbored various MGEs, primarily plasmids, and demonstrated the ability to protect their DNA cargo from degradation and facilitate horizontal gene transfer, including the transmission of resistance genes. In conclusion, this study reveals widespread presence of BMVs carrying ARGs and potential virulence genes in swine farm feces, soil, and fecal wastewater. These findings not only provide new insights into the role of extracellular DNA in the environment but also highlight concerns regarding the gene transfer potential mediated by BMVs and associated health risks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Pathogenic Escherichia coli (PEC) are important foodborne bacteria that can cause severe illness in humans. The PECs thrive within the intestines of humans as well as animals and may contaminate multiple ecosystems, including food and water, via faecal transmission. Abattoir and farm employees are at high risk of PEC exposure, which could translate to community risk through person-to-person contact. To determine the epidemiology and resistome of PECs in Gauteng and Limpopo provinces of South Africa, 198 swine faecal samples, 220 poultry cloacal swabs, 108 human hand swabs, 11 run-off water samples from abattoirs and farms were collected from four swine and five poultry commercial abattoirs and two swine farms. One effluent sample each was collected from four wastewater treatment plants (WWTP) and a tertiary hospital setting. Phenotypic and genotypic techniques were used including polymerase chain reaction, pulsed-field gel electrophoresis (PFGE) and whole genome sequencing (WGS). Results showed EHEC and EPEC prevalence was 4.1 % (22/542) and 20.8 % (113/542), respectively, with the O26 serogroup detected the most in PEC isolates. According to the PFGE dendrogram, isolates from poultry, human hand swabs and run-off water clustered together. Diverse virulence factors such as the novel stx2k subtype and eae genes were detected among the 36 representative PEC isolates according to WGS. The results showed that 66.7 % (24/36) of sequenced PECs presented with multi-drug resistance (MDR) to β-lactamase 13.9 % (5/36), aminoglycoside 61.1 % (22/36), tetracycline 41.7 % (15/36) and quinolones 38.9 % (14/36). No colistin nor carbapenem resistance was detected. Sequence types (STs) associated with MDR in this study were: ST752, ST189, ST206, ST10, ST48 and ST38. The findings highlight the threat of zoonotic pathogens to close human contacts and the need for enhanced surveillance to mitigate the spread of MDR foodborne PECs., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Samuel T. Ogundare reports financial support was provided by National Research Foundation. Anthony M Smith reports financial support was provided by United Kingdom Department of Health and Social Care. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Furanoids are a class of contaminants prevalent in both airborne and occupational environments, with potential health implications through inhalation, oral ingestion, and skin penetration. Given their diminutive molecular size, there is a presumption that furanoids can readily permeate the skin. To systematically explore this presumption, we investigated the skin absorption and toxicity of a series of furans (furfuryl alcohol, furfuryl acetate, furfural, methyl 2-furoate, and 5-methylfurfural) using in silico, in vitro, and in vivo models. The in vitro permeation test (IVPT) from neat and aqueous suspension (5 mM) of furans demonstrated a facile absorption through pig and nude mouse skins. The lipophilicity of furans significantly influenced skin deposition, with higher lipophilicity displaying greater deposition. However, an opposing trend emerged in the receptor compartment accumulation. In barrier-defective skin simulating atopic dermatitis (AD) and psoriasis, enhanced deposition occurred with more hydrophilic furans but not with the more lipophilic ones. In the cell-based study, furanoids induced the proliferation of keratinocytes and skin fibroblasts except for the compounds with the aldehyde group (furfural and 5-methylfurfural). Both furfuryl acetate and 5-methylfurfural activated keratinocytes via the overexpression of COX-2 and PGE 2 by 1.5‒2-fold. This stimulation involved the mitogen-activated protein kinase (MAPK) signaling pathway. For the in vivo mouse skin treatment, we selected furfuryl acetate (hydrophilic) and 5-methylfurfural (lipophilic). Both furans showed different patterns of skin lesions, where repeated application of furfuryl acetate caused epidermal hyperplasia and scaling, while 5-methylfurfural predominantly evoked skin inflammation and barrier disintegration. Toxicokinetics analysis revealed a higher plasma concentration of topically applied furfuryl acetate than that of the 5-methylfurfural (5.04 versus 2.34 nmol/ml), resulting in the mild injury of furfuryl acetate-treated peripheral organs. Conversely, no notable adverse effects on organs were observed for the 5-methylfurfural. This study established the relationship between cutaneous absorption and the toxicity of furans following skin exposure., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This study presents a method utilizing solid-phase microextraction Arrow (SPME Arrow) combined with ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for the selective detection of three veterinary drugs-thiabendazole, sulfamethazine, and clenbuterol-in milk and pork. Two-dimensional metal-organic framework nanosheets (2D-MOFs) were employed as coating materials for the SPME Arrow. Three types of 2D-MOFs (Ni, Mn, and Co based) were synthesized and characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and a physical adsorption analyzer. The 2D-MOF coatings were fabricated using the electrospinning technique, with polyacrylonitrile (PAN) serving as the binder. Comparative analysis of the three 2D-MOF coatings revealed that 2D-Ni-MOF was the optimal coating material for the SPME Arrow. Optimization of the coating preparation conditions and SPME procedures included determining the optimal mass ratio of 2D-Ni-MOF to PAN, electrospinning time, and extraction and desorption parameters. Equilibrium extraction was achieved within 60 min, and desorption was completed within 30 min. Subsequently, the 2D-Ni-MOF-SPME Arrow-UPLC-Q-TOF-MS method was established and validated under optimal conditions, demonstrating high precision with inter-day precision ranging from 3.8 % to 9.5 % and intra-day precision ranging from 5.1 % to 11.5 %. The reusability study indicated that the extraction performance of the new SPME Arrow remained consistent after 90 adsorption-desorption cycles. The method exhibited linearity in milk and pork over the ranges of 0.002-5 μg L -1 and 0.01-5 μg L -1 , respectively. The detection limits in milk and pork were 0.001-0.004 μg L -1 and 0.003-0.007 μg L -1 , respectively. This method demonstrated excellent applicability for determining residues of the three veterinary drugs in milk and pork., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Hangzhen Lan reports financial support was provided by Ningbo University. Hangzhen Lan reports financial support was provided by One Health Interdisciplinary Research Project of Ningbo University. Hangzhen Lan reports financial support was provided by J National Natural Science Foundation of China. Hangzhen Lan reports financial support was provided by Key R&D Program of Zhejiang. Hangzhen Lan reports financial support was provided by Natural Science Foundation of Zhejiang Province. Hangzhen Lan reports financial support was provided by Science and Technology Programs of Ningbo., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sulfachloropyridazine (SCP) is a common sulfonamide antibiotic pollutant found in animal excreta. Finding highly efficient degrading bacterial strains is an important measure to reduce SCP antibiotic pollution. Although some strains with degradation capabilities have been screened, the degradation pathways and biotransformation mechanisms of SCP during bacterial growth are still unclear. In this study, a strain capable of efficiently degrading SCP, named Bacillus sp. DLY-11, was isolated from pig manure aerobic compost. Under optimized conditions (5 % Vaccination dose, 51.5 ℃ reaction temperature, pH=7.92 and 0.5 g/L MgSO 4 ), this strain was able to degrade 97.7 % of 20 mg/L SCP within 48 h. Through the analysis of nine possible degradation products (including a new product of 1,4-benzoquinone with increased toxicity), three potential biodegradation pathways were proposed. The biodegradation reactions include S-N bond cleavage, dechlorination, hydroxylation, deamination, methylation, sulfur dioxide release, and oxidation reactions. This discovery not only provides a new efficient SCP-degrading bacterial strain but also expands our understanding of the mechanisms of bacterial degradation of SCP, filling a knowledge gap. It offers important reference for the bioremediation of antibiotic pollutants in livestock and poultry farming., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Green agriculture faced challenges due to the shortage of efficient cadmium (Cd)-resistant plant growth-promoting bacteria (CdR-PGPB) and their low survival rate and activity during application. In this study, a diverse range of efficient CdR-PGPB were isolated from the rhizosphere soil of Desmodium elegans, especially those with high phosphate-solubilizing capabilities (272.87-450.45 mg L -1 ). Two highly efficient CdR-PGPB namely, XH1 and XH3 were loaded on to rice husk biochar (RHB) and pig bone biochar (PBB), labelled as RHBM and PBBM respectively. This study aimed to explore their effectiveness and mechanisms in promoting maize growth in a Cd-contaminated planting system. Results showed that PBBM performed best among all treatments. It significantly decreased soil phytoavailable Cd by 53.19 % and Cd content in maize shoot by 85.89 %. It also increased soil available phosphorus by 145.72 %, soil alkaline phosphatase activity by 76.34 %, maize shoot/root biomass by 47.06 %/67.98 %, Chlorophyll (a/b) content by 66.80 %/134.13 % and peroxidase activity by 171.96 %. These results were achieved through the synergistic action of efficient CdR-PGPB and PBB. Therefore, PBBM proved to be a promising and innovative application technique for sustainable agricultural development in Cd-contaminated farmland ecosystems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Porcine epidemic diarrhea virus (PEDV) and rotavirus has posed a significant threat to the pig industry annually across different nations, resulting in huge economic losses. The frequent co-infection of these two viruses in clinical settings complicates the process of differential diagnoses. Rapid and accurate detection of PEDV and rotavirus is in great demand for timely diarrhea disease prevention and control. In this study, tris stabilized AuNPs were prepared and a sensitive lateral flow immunoassay (LFIA) sensor was developed for the simultaneous and rapid detection of PEDV and rotavirus on site. After the system optimization, the established LFIA can simultaneously identify PEDV and rotavirus with limits of detection (LOD) of 1.25 × 10 3 TCID 50 mL -1 and 3.13 × 10 2 pg mL -1 , respectively. When applying for clinical samples, the LFIA show a concordance of 95 % and 100 % to reverse transcript polymerase chain reaction (RT-PCR) for PEDV and rotavirus respectively. Therefore, this LFIA can qualitatively detect PEDV and rotavirus in 18 min with high sensitivity and accuracy without any sophisticated equipment and operation, making it a promising candidate for the early diagnosis of PEDV or/and rotavirus diarrhea on site., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Hailan Chen reports financial support was provided by Bama County for Talents in Science and Technology. Qianlian Su reports financial support was provided by Project for Enhancing Young and Middle-aged Teacher's Research Basis Ability in Universities in Guangxi. Hui You reports financial support was provided by Guangxi Bagui Scholar Program. Jiakang He reports financial support was provided by Guangxi Key R&D Program Project.]., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: To investigate if fentanyl induces immobility through activation of the serotonergic 5HT 1A receptor, by using the 5HT 1A -antagonist robalzotan., Study Design: A prospective, blinded, randomized, two-group study., Animals: A group of 12 mixed-breed pigs aged 71-79 days., Methods: The motor response to clamping a claw was assessed in isoflurane-anaesthetized pigs at baseline, then fentanyl was infused intravenously (IV) for 40 minutes and clamping was repeated. The infusion started at 20 μg kg -1 hour -1 and was increased by 60% until fentanyl produced immobility, defined as no motor response for 60 seconds. Subsequently, either robalzotan (1 mg kg -1 ) or the same volume of saline was injected IV and clamping was repeated. The change in response was compared with Fisher's exact test. Mean arterial blood pressure (MAP) and heart rate (HR) were extracted for 2 minutes before and after 60 seconds of clamping, and the differences compared with a Wilcoxon signed-rank test. Dynamic respiratory compliance was calculated at baseline and after fentanyl; p < 0.05., Results: Baseline clamping produced a motor response within 5 seconds. This was abolished by fentanyl. Robalzotan or saline did not alter this (p = 0.45). As a response to clamping, MAP and HR changed with median (range) -0.5 (-4.4 to 22.2) mmHg and -1 (-7 to 1.5), respectively, where HR changed significantly (p = 0.039). The 95% confidence interval for the effect size of fentanyl upon dynamic compliance was -3.25 to -1.65 mL cmH 2 O -1 ., Conclusions and Clinical Relevance: No indication was found for the 5HT 1A receptor to be involved in fentanyl-induced reduction of the motor response to claw clamping. The decreased compliance after fentanyl could suggest onset of chest wall rigidity., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objective: To collect validity evidence for the use of the Anastomosis Objective Structured Assessment of Technical Skills (A-OSATS) instrument, which has been developed to evaluate performance of a minimally invasive side-to-side bowel anastomosis with hand-sewn common enterotomy., Design: Residents performed a robotic ileocolic anastomosis simulation on an ex vivo porcine model. Faculty scored each resident with the A-OSATS and performed a provocative leak test on the completed anastomoses. Residents were reassessed on the sewing sub-score 1 month later. Data were compared with parametric and nonparametric analysis., Setting: Single academic general surgery residency PARTICIPANTS: PGY-4 and -5 general surgery residents (n = 17) RESULTS: PGY-5s performed better than PGY-4s in repeat A-OSATS sewing sub-score (mean 55/55 ± 0 vs 43 ± 4.9, p < 0.001) and time to complete (minutes, mean 14.5 ± 4.9 vs 21.2 ± 3.9, p = 0.01). There was a strong correlation between A-OSATS score and time (r = -0.67, p = 0.005). For the initial assessment, there was no significant difference in mean A-OSATS score between anastomoses that leaked and those that did not leak (137.3 ± 14.5 vs 150.1 ± 11.2, p = 0.098), but on repeat assessment, intact anastomoses had a higher mean A-OSATS sewing sub-score than those that leaked (52.2 ± 4.7 vs 39 ± 3.5, p = 0.007). There was no significant difference between initial A-OSATS score and repeat score (p = 0.14)., Conclusions: We provide extrapolative validity evidence for the A-OSATS instrument by comparing A-OSATS score to time to sew, provocative leak test, and discrimination between PGY-4s and PGY-5s. Generalizability validity evidence is provided by test-retest reliability. Further refinement is needed for the A-OSATS tool to be used for high-stakes entrustment decisions in resident-performed robotic ileocolic anastomoses., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Porcine reproductive and respiratory syndrome (PRRS) caused by porcine reproductive and respiratory syndrome virus (PRRSV) causes considerable economic losses to the global swine industry every year and seriously hinders the healthy development of this industry. Although tremendous efforts have been made over the past 30 years toward the development of prevention and control strategies against PRRSV infection, to date, treatments with proven efficacy have yet to be available due to our incomplete understanding of the molecular basis and complexity of the infection machinery. This review systematically discusses recent advances in the research and development of anti-PRRSV therapies targeting different stages of the viral life cycle. Furthermore, this review puts forward novel intervention targets and research approaches based on our in-depth exploration of virus-host interactions and the latest biological technologies, which have the potential to complement or transform current anti-PRRSV strategies and become breakthrough points for the control of PRRS in the future., Competing Interests: Declaration of competing interest All authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Porcine deltacoronavirus (PDCoV) poses a significant threat to both the pig industry and public safety, and has recently been identified in humans. Currently, there are no commercially available vaccines or antiviral treatments for PDCoV. In this study, recombinant porcine interferon δ8 (rINF-δ8) expressed by the HEK 293F expression system was used to evaluated its antiviral activity against PDCoV both in vitro and in vivo. Results demonstrated that rIFN-δ8 displayed non-toxic to ST cells and primary PAMs, and effectively inhibited PDCoV replication in a dose-dependent manner in vitro, with complete suppression of virus replication at a concentration of 2 μg/ml. Treatment of piglets with two doses of 25 μg/kg of rIFN-δ8 reduced clinical symptoms, decreased virus shedding, alleviated intestinal damage, and lowered the viral load in the jejunum and ileum. Furthermore, the levels of interferon-stimulated genes (ISGs) such as Viper, Mx1, ISG15, IFIT1, OSA, and IFITM1 were significantly increased both in vitro and in vivo, with elevated ISG levels sustained for at least 3 days in vivo. These findings suggest that rIFN-δ8 has the potential to serve as an effective antiviral agent for preventing PDCoV in pigs in the future., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Porcine epidemic diarrhea virus (PEDV) causes enormous economic losses to the pork industry, and its extensive cell tropism poses a substantial challenge to public health and safety. However, the invasion mechanisms and relevant host factors of PEDV remain poorly understood. In this study, we identified 422 differentially expressed genes related to PEDV infection through transcriptome analysis. Among these, Annexin A2 (ANXA2), Prohibitin-2 (PHB2), and Caveolin-2 (CAV2) were identified through screening and verifying as having a specific interaction with the PEDV S protein, and positive regulation of PEDV internalization was validated by siRNA and overexpression tests. Subsequently, using host membrane protein interaction networks and co-immunoprecipitation analysis, we found that ANXA2 PHB2 or CAV2 directly interact with Rab11a. Next, we constructed a pseudovirus model (LV-PEDV S-GFP) to further confirm that the downregulation of Rab11a could promote PEDV invasion. In detail, ANXA2, PHB2, or CAV2 promoted PEDV invasion via downregulating Rab11a. Furthermore, we showed that the S-protein fusion peptide (FP) was sufficient for S-protein interaction with ANXA2, PHB2, CAV2, and Rab11a, and the addition of exogenous GTP could regulate the efficiency of PEDV invasion. Collectively, ANXA2, PHB2, or CAV2 influenced the membrane fusion of PEDV with host cells through the host restriction factor Rab11a. This study could be targeted for future research to develop strategies for the control of PEDV., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Objectives: To investigate the correlation between the serum levels of 25(OH)D and the resorption of the alveolar bone walls and regeneration of the alveolar space after tooth extraction., Methods: 14 adults in need of extraction of hopeless teeth were enrolled. An intraoral digital impression was performed, and each patient was tested to assess serum vitamin D levels. Subsequently, extraction of teeth and contextual guided bone regeneration was performed using porcine origin graft material and a resorbable collagen membrane to covert the defect. After 4 months, an impression was taken, and the model was scanned using a professional scanner for lab. At the same time, a cone beam computed tomography was performed to plan implant insertion through fully digital computer guided surgery. Bone was collected to perform histological and histomorphometric analysis. Pre and postoperative scans were compared using a specific software to estimate the volumetric changes. Tests were applied to investigate the relationship between the different predictor variables and the outcome variables., Results: 14 patients were divided in 3 groups depending on the serum Vit-D levels, identifying three ranges corresponding to low (lower than 20), medium (between 20 and 30), and optimal vitamin D levels (higher than 30). Volumetric contraction after extraction was observed for all patients, without any significant difference between the groups. Focusing on the post-extraction regeneration, patients belonging to the group with lower levels of Vit-D displayed lower and more disorganized levels of bone. Immunohistochemistry analysis showed that Col1A1 and Osteocalcin had no physiological alteration. Osteopontin could be identified near the external surface of bone tissue granules. Runx2 signals were detected near the margins of bone trabeculae., Conclusions: Serum vit-D levels do not appear to influence the extent of post-extraction bone contraction; on the contrary, they seem to influence the post-extraction regeneration., Clinical Significance: Vit D serum levels may influence the regenerative aspect during post-extraction turn-over. This might suggest controlling and (in case of low levels) recommend Vit D supplement in the patient diet in case of extraction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Ballistic gelatin has been extensively used in ballistics research for decades, but calibration standards were established on limited datasets, and only few studies have attempted to recreate these experiments with biological tissues. Recent studies have demonstrated better biofidelity with 20% ordnance ballistic gelatin, but researchers have discredited the use of synthetic gelatin claiming different behavior than ordnance gelatin. To investigate the use of synthetic clear gelatin as an acceptable surrogate of biological tissue, depth of penetration was compared between low-velocity impacts of various projectiles into porcine tissue (n = 192), post-mortem human subjects (n = 29), and Clear Ballistics synthetic gelatin (n = 39). The predicted depth of penetration of the 0.177" steel BB (38.1 mm) was consistent with the manufacturer's calibration standard (31.75-44.45 mm) and within calibration bounds of recently proposed empirical equations. Compared to impacts in biological tissue, synthetic gelatin demonstrated the least variability in depth of penetration (R 2  = 0.96). Using ANCOVA, velocity was a significant covariate (p < 0.001), and there were no significant differences in normalized depth of penetration over density between porcine tissue, post-mortem human subjects, and 20% synthetic gelatin (p = 0.22). Ultimately, this study confirmed the use of 20% synthetic gelatin as an acceptable tissue simulant using standard calibration methods for use in future ballistic studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

An intelligent pH response indicator film is an easy-to-use device for the real-time monitoring of meat freshness during transport and storage. Therefore, a novel pH-sensitive anthocyanin indicator film composed of polyvinyl alcohol-blueberry anthocyanin (BA)-2-hydroxypropyltrimethyl ammonium chloride chitosan (HACC) called PAH-2.0 with 1.2 mg/mL HACC to monitor meat freshness using HACC as the colorimetric enhancer has been developed. BA and HACC were mixed and immobilized in the polyvinyl alcohol matrix by hydrogen bonds, as confirmed via Fourier-transform infrared spectroscopy and X-ray diffraction. The inclusion of HACC improved the color stability and antioxidant and antibacterial properties of the PAH-2.0 film. When applied to pork for freshness monitoring at 4 °C, three freshness stages, including fresh, sub-fresh, and spoiled, could be clearly distinguished based on the color variations of the PAH-2.0 film. The distinct hierarchical color change from purple to blue-violet and finally to grayish-blue was highly correlated with the indicators of pork freshness: pH values, total volatile basic nitrogen, and total viable count. This study provides a simple and promising approach for fabricating meat freshness indicator films with high color recognition accuracy, thereby offering new possibilities for visual meat freshness monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hemoadhican (HD) is an exopolysaccharide with a branched structure that has been reported for its high hemostatic ability. In this study, a HD-based hemostatic sponge was prepared through ultrasonic dissolution and freeze-drying without using any cross-linking agent. The sponge could spontaneously cross-link using hydrogen bonds to form adhesive mud within 3 s upon contact with blood. This sponge-mud mixture adhered tightly to the wound tissue, forming a pressure-resistant physical barrier that captures and locks in blood cells and platelets. Simultaneously, the hydrophobic methyl groups of HD sponges repel blood inwardly, effectively sealing the wound. The brush-like structure of HD molecules was suspected to penetrate wet tissues through topological entanglement, thereby enhancing wet adhesion. Compared with gauze and gelatin sponges, HD sponges achieved more effective hemostasis in animal models using rat and rabbit femoral arteries. In particular, HD sponges showed excellent hemostasis in heparin-induced hemorrhage models in mice and pigs. The in vivo experiment demonstrated the excellent biosafety of the HD sponge. Conclusively, the HD sponge is a safe and efficient rapid hemostatic material that is expected to become an alternative material for clinical hemostatic procedures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Objective: To determine if in vivo cryoneurolysis inhibits ex vivo compound action potential (CAP) conduction in the porcine saphenous nerve and if this occurs rapidly enough to justify performing the technique before stifle surgery., Study Design: Blinded, controlled, randomized, preclinical study., Animals: A group of eight healthy, 8 weeks old, intact, female pigs anesthetized for an unrelated terminal study., Methods: Both saphenous nerves of each pig were exposed surgically, and 15 mm of a 20 gauge, closed-tip, commercial cryoneurolysis cannula were inserted cranial to each nerve within the neurovascular fascial sheath along its long axis. The cannula was only actuated on one limb, according to random allocation. Nerves were excised within 15 minutes of actuation and underwent testing in a nerve conduction chamber, where stimulus voltage was increased sequentially (from 0.1 to ≤ 1.9 V). An anesthesiologist blinded to treatment viewed recordings of time versus voltage for each nerve and answered 'yes' or 'no' when asked if an evoked CAP was observed. Fisher's exact test evaluated the incidence of CAP conduction between groups (p < 0.05 considered significant). Nerves were submitted for hematoxylin and eosin staining for blinded histopathological examination., Results: A CAP was conducted in 8/8 and 1/8 of the control and treated nerves, respectively (p = 0.001). Maximal responses in control nerves were 1.92 ± 0.19 mV (mean ± standard error). In the single treated nerve that conducted a CAP, the maximal CAP amplitude was 0.4 mV, lower than the lowest maximal CAP (1.19 mV) in the control nerves. All control nerves were histologically normal, and all treated nerves displayed mild perivascular and perineural inflammation (cuffs of lymphocytes, plasma cells and eosinophils, and edema)., Conclusions and Clinical Relevance: The rapid inhibition of CAP conduction warrants clinical investigation of saphenous cryoneurolysis for both intraoperative antinociception and postoperative analgesia in pigs undergoing experimental stifle surgery., (Copyright © 2024 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Early diagnosis of pregnancy is directly related to cost-effective livestock production. We produced a rat monoclonal antibody (mAb) against synthesized porcine early pregnancy factor (pEPF) using conventional hybridoma technology and used it as a tool for the detection of early pregnancy in Duroc sows. The rat pEPF-mAb showed reactivity to uterine tissues of pregnant sows 20 or 30 days post-mating (day 0 defined as the day of mating) and non-pregnant sows (confirmed signs of estrus) in western blotting. Immunohistochemical analysis confirmed that pEPF was located in the stromal and grand epithelial tissues of pregnant sows 20 or 30 days post-mating. In the enzyme-linked immunosorbent assay, pEPF expression in urine and blood showed similar results, with the highest expression observed in pregnant sows 20 days post-mating, whereas there was no significant difference in expression levels between non-pregnant sows and pregnant sows 30 days post-mating. The pEPF-mAb-based pregnancy diagnostic kit can be applied to pig urine samples non-invasively collected at 20 days post-mating with 70 % accuracy. Further improvements to the kit's diagnostic performance may lead to substantial benefits for the swine industry, facilitating more efficient and accurate reproductive management., Competing Interests: Declaration of competing interest No potential conflict of interest was reported by the author(s)., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Calcium ions (Ca 2+ ) regulate cell proliferation and differentiation and participate in various physiological activities of cells. The calcium transfer protein inositol 1,4,5-triphosphate receptor (IP 3 R), located between the endoplasmic reticulum (ER) and mitochondria, plays an important role in regulating Ca 2+ levels. However, the mechanism by which IP 3 R1 affects porcine meiotic progression and embryonic development remains unclear. We established a model in porcine oocytes using siRNA-mediated knockdown of IP 3 R1 to investigate the effects of IP 3 R1 on porcine oocyte meiotic progression and embryonic development. The results indicated that a decrease in IP 3 R1 expression significantly enhanced the interaction between the ER and mitochondria. Additionally, the interaction between the ER and the mitochondrial Ca 2+ ([Ca 2+ ] m ) transport network protein IP 3 R1-GRP75-VDAC1 was disrupted. The results of the Duolink II in situ proximity ligation assay (PLA) revealed a weakened pairwise interaction between IP 3 R1-GRP75 and VDAC1 and a significantly increased interaction between GRP75 and VDAC1 after IP 3 R1 interference, resulting in the accumulation of large amounts of [Ca 2+ ] m . These changes led to mitochondrial oxidative stress, increased the levels of reactive oxygen species (ROS) and reduced ATP production, which hindered the maturation and late development of porcine oocytes and induced apoptosis. Nevertheless, after treat with [Ca 2+ ] m chelating agent ruthenium red (RR) or ROS scavenger N-acetylcysteine (NAC), the oocytes developmental abnormalities, oxidative stress and apoptosis caused by Ca 2+ overload were improved. In conclusion, our results indicated IP 3 R1 is required for meiotic progression and embryonic development by regulating mitochondrial calcium and oxidative damage., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The incidence of "acid attacks" (vitreolage) is a global concern, with those affected often receiving lifelong medical care due to physical and psychological damage. The purpose of this study was to evaluate the effectiveness of several emergency skin decontamination approaches against concentrated (>99 %) sulphuric acid and to identify the effective window of opportunity for decontamination. The effects of four decontamination methods (dry, wet, combined dry & wet and cotton cloth) were assessed using an in vitro diffusion cell system containing dermatomed porcine skin. Sulphuric acid (H 2 SO 4 ) was applied to the skin with decontamination protocols performed at 10 s, 30 s, 8 min, and 30 min post exposure. Skin damage was quantified by tritiated water ( 3 H 2 O) penetration, receptor fluid pH and photometric stereo imaging (PSI), with quantification of residual sulphur (by SEM-EDS) to determine overall decontamination efficiency. Skin translucency (quantified by PSI) demonstrated a time-dependent loss of dermal tissue integrity from 10 s. Quantification of dermal sulphur content confirmed the rapid (exponential) decrease in decontamination efficiency with time. The pH of the water effluent indicated complete neutralisation of acid from the skin surface after 90 s of irrigation. Wet decontamination (either alone or immediately following dry decontamination) was the most effective intervention evaluated, although no decontamination technique was statistically effective after 30 s exposure to the acid. These data demonstrate the time-critical consequences of dermal exposure to concentrated sulphuric acid: we find no practical window of opportunity for acid decontamination, as physical damage is virtually instantaneous., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Ltd.)