Randomised Controlled Trial on Prophylaxis of Mastitis-Metritis-Agalactia Syndrome in Swine using Caulophyllum Logoplex and Lachesis Logoplex.

BACKGROUND: Mastitis-metritis-agalactia (MMA) syndrome occurs in the first days post-partum and causes piglet losses mainly due to malnutrition. One possibility for prophylaxis of MMA is via homeopathy. In this veterinary study, the effectiveness of a prophylactic administration of homeopathic remedies for the prevention of the occurrence of MMA in swine was evaluated. METHODS: In a randomised and blinded study, 60 sows were examined. Sows were randomly distributed in two groups: the experimental group (CL/LL) received a prophylactic administration of the complex homeopathic remedies Caulophyllum Logoplex and Lachesis Logoplex, and the placebo group was administered a sodium chloride (NaCl) solution in the same injection scheme as the experimental group. Clinical signs of MMA, behavioural changes, as well as production parameters, were recorded beginning with the day of farrowing until 5 days post-partum. RESULTS: The treatment group showed no significant effect on the occurrence of MMA in sows (CL/LL: 56.67% MMA positive sows; NaCl: 53.53% MMA positive sows). Treatment group had also no significant effect on health parameters (vaginal discharge, raised rectal temperature, shortage of milk) or behavioural parameters (impaired feeding behaviour and impaired general condition). For the production parameter average weight gain, statistically significant effects in the treatment group were detected. CONCLUSIONS: Prophylaxis with the homeopathic remedies Caulophyllum Logoplex and Lachesis Logoplex showed neither an improvement in MMA prevention nor an improvement in health parameters or behavioural traits in the present herd of sows.

A Comparative Transcriptomic Analysis Reveals That HSP90AB1 Is Involved in the Immune and Inflammatory Responses to Porcine Deltacoronavirus Infection.

PDCoV is an emerging enteropathogenic coronavirus that mainly causes acute diarrhea in piglets, seriously affecting pig breeding industries worldwide. To date, the molecular mechanisms of PDCoV-induced immune and inflammatory responses or host responses in LLC-PK cells in vitro are not well understood. HSP90 plays important roles in various viral infections. In this study, HSP90AB1 knockout cells (HSP90AB1KO) were constructed and a comparative transcriptomic analysis between PDCoV-infected HSP90AB1WT and HSP90AB1KO cells was conducted using RNA sequencing to explore the effect of HSP90AB1 on PDCoV infection. A total of 1295 and 3746 differentially expressed genes (DEGs) were identified in PDCoV-infected HSP90AB1WT and HSP90AB1KO cells, respectively. Moreover, most of the significantly enriched pathways were related to immune and inflammatory response-associated pathways upon PDCoV infection. The DEGs enriched in NF-κB pathways were specifically detected in HSP90AB1WT cells, and NF-κB inhibitors JSH-23, SC75741 and QNZ treatment reduced PDCoV infection. Further research revealed most cytokines associated with immune and inflammatory responses were upregulated during PDCoV infection. Knockout of HSP90AB1 altered the upregulated levels of some cytokines. Taken together, our findings provide new insights into the host response to PDCoV infection from the transcriptome perspective, which will contribute to illustrating the molecular basis of the interaction between PDCoV and HSP90AB1.

Supplementation with Exogenous Catalase from Penicillium notatum in the Diet Ameliorates Lipopolysaccharide-Induced Intestinal Oxidative Damage through Affecting Intestinal Antioxidant Capacity and Microbiota in Weaned Pigs.

The present study aimed to explore the protective effects of exogenous catalase (CAT) from microorganisms against lipopolysaccharide (LPS)-induced intestinal injury and its molecular mechanism in weaned pigs. Fifty-four weaned pigs (21 days of age) were randomly allocated to CON, LPS, and LPS+CAT groups. The pigs in CON and LPS groups were fed a basal diet, whereas the pigs in LPS+CAT group fed the basal diet with 2,000 mg/kg CAT supplementation for 35 days. On day 36, six pigs were selected from each group, and LPS and LPS+CAT groups were administered with LPS (50 µg/kg body weight). Meanwhile, CON group was injected with an equivalent amount of sterile saline. Results showed that LPS administration damaged intestinal mucosa morphology and barrier. However, CAT supplementation alleviated the deleterious effects caused by LPS challenge through enhancing intestinal antioxidant capacity which was benefited to decrease proinflammatory cytokines concentrations and suppress enterocyte apoptosis. Besides, LPS-induced gut microbiota dysbiosis was significantly shifted by CAT through decreasing mainly Streptococcus and Escherichia-Shigella. Our study suggested that dietary supplemented with 2,000 mg/kg catalase was conducive to improve intestinal development and protect against LPS-induced intestinal mucosa injury via enhancing intestinal antioxidant capacity and altering microbiota composition in weaned pigs. IMPORTANCE Exogenous CAT derived from microorganisms has been widely used in food, medicine, and other industries. Recent study also found that exogenous CAT supplementation could improve growth performance and antioxidant capacity of weaned pigs. However, it is still unknown that whether dietary exogenous CAT supplementation can provide a defense against the oxidative stress-induced intestinal damage in weaned pigs. Our current study suggested that dietary supplemented with 2,000 mg/kg CAT was conducive to improve intestinal development and protect against LPS-induced intestinal mucosa injury via enhancing intestinal antioxidant capacity and altering microbiota composition in weaned pigs. Moreover, this study will also assist in developing of CAT produced by microorganisms to attenuate various oxidative stress-induced injury or diseases.

Vaginal microbiota differences associated with pelvic organ prolapse risk during late gestation in commercial sows†.

During the last decade, sow mortality due to pelvic organ prolapse (POP) has increased. To better understand the biology associated with POP, sows were phenotypically assessed and assigned a perineal score (PS) based on presumed POP risk and categorized as PS1 (low), PS2 (moderate), or PS3 (high). The study objective was to identify changes in sow vaginal microbiota that may be associated with POP. The hypothesis is that vaginal microbiota differs between sows with variable risk for POP, and changes in microbiota during late gestation exist between sows with differing risk. Of the 2864 sows scored during gestation week 15, 1.0, 2.7, and 23.4% of PS1, PS2, and PS3 sows, respectively, subsequently experienced POP. Vaginal swabs subjected to 16S rRNA gene sequencing revealed differences in community composition (Bray-Curtis; P < 0.05) and individual operational taxonomic unit (OTU) comparisons between vaginal microbiota of PS1 and PS3 sows at gestation week 15. Further, differences (P < 0.05) in community composition and OTUs (Q < 0.05) were observed in PS3 sows that either did or did not subsequently experience POP. Differences in community structure (alpha diversity measurements; P < 0.05), composition (P < 0.05), and OTUs (Q < 0.05) were observed in gestation week 12 sows scored PS1 compared to week 15 sows scored PS1 or PS3, suggesting that sow vaginal microbiota shifts during late gestation differently as POP risk changes. Collectively, these data demonstrate that sows with greater POP risk have unique vaginal microflora, for which a better understanding could aid in the development of mitigation strategies.

Porcine ear necrosis.

Porcine ear necrosis (PEN) is a condition that mainly occurs in intensive pig production systems and mostly affects piglets after weaning. The syndrome manifests itself with lesions on the pinna, which can heal or become more severe resulting in partial loss of the ear. The pathogenesis of the condition is not fully known. Three different hypotheses for the development of PEN are described in this review: (1) damage of the epidermis due to Staphylococcal exfoliative toxins; (2) occlusion of small blood vessels; and (3) ear biting with subsequent ß-hemolytic streptococcal infection. Risk factors have not been completely elucidated, but viral and bacterial infections, and husbandry factors such as environment, housing conditions and management, have been suggested. It is also possible that some cases are due to a combination of these factors. The role of parasitic infestations has been not investigated. Due to bacterial involvement, severely affected pigs can be treated with antimicrobials. Control and preventive measures should focus on reducing potential risk factors by implementing herd immunization, as well as improvement of sanitary conditions, feed quality (with respect to mycotoxin contamination), management (appropriate stocking density), and environmental conditions (e.g. number of drinkers and feeders and/or optimal ventilation). Further research is needed to better understand the precise etiology and pathogenesis of PEN, so that risk factors can be identified and more targeted control measures can be implemented.

Swine Enteric Coronaviruses (PEDV, TGEV, and PDCoV) Induce Divergent Interferon-Stimulated Gene Responses and Antigen Presentation in Porcine Intestinal Enteroids.

Swine enteric coronaviruses (SECoVs) including porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV), account for the majority of lethal watery diarrhea in neonatal pigs and pose significant economic and public health burdens in the world. While the three SECoVs primarily infect intestinal epithelia in vivo and cause similar clinical signs, there are evident discrepancies in their cellular tropism and pathogenicity. However, the underlying mechanisms to cause the differences remain unclear. Herein, we employed porcine enteroids that are a physiologically relevant model of the intestine to assess the host epithelial responses following infection with the three SECoVs (PEDV, TGEV, and PDCoV). Although SECoVs replicated similarly in jejunal enteroids, a parallel comparison of transcriptomics datasets uncovered that PEDV and TGEV infection induced similar transcriptional profiles and exhibited a more pronounced response with more differentially expressed genes (DEGs) in jejunal enteroids compared with PDCoV infection. Notably, TGEV and PDCoV induced high levels of type I and III IFNs and IFN-stimulated gene (ISG) responses, while PEDV displayed a delayed peak and elicited a much lesser extent of IFN responses. Furthermore, TGEV and PDCoV instead of PEDV elicited a substantial upregulation of antigen-presentation genes and T cell-recruiting chemokines in enteroids. Mechanistically, we demonstrated that IFNs treatment markedly elevated the expression of NOD-like receptor (NLR) family NLRC5 and major histocompatibility complex class I (MHC-I) molecules. Together, our results indicate unique and common viral strategies for manipulating the global IFN responses and antigen presentation utilized by SECoVs, which help us a better understanding of host-SECoVs interactions.

Flooding on Beef and Swine Farms: A Scoping Review of Effects in the Midwestern United States.

BACKGROUND: Increasing uncertainty and variability in weather due to climate change puts enormous stress on the sustainability of agricultural communities in several parts of the continental United States. Rural agriculture-based communities, such as those in North Carolina, Nebraska, and Illinois are particularly vulnerable to the impacts of flooding. These extreme weather events affect many animal facilities, and flooding can cause long-term impacts on animal health and productivity. OBJECTIVES: This scoping review investigated the potential short- and long-term ramifications of flooding on beef cattle and swine farms with theoretical locations in the Midwestern U.S. The goal of this review is to create a concept map to identify research gaps related to efforts to support those animals and peoples ravaged by floods. This was accomplished by performing a scoping review using search terms "beef OR swine AND flood" to assess direct effects and then "beef OR swine AND direct effect" to evaluate secondary and tertiary effects. RESULTS: Our findings were based on a total of 89 peer-reviewed publications, with 50 publications relating to beef cattle, 24 relating to swine, and 15 containing information applicable to both. On beef farms, the effects of flooding can be broad and long-lasting. The short-term risks of flooding are reasonably well-understood, including the comingling of unfamiliar cattle, nutritional restriction, and disease transmission. However, long-term impacts, including potential effects on reproduction, nutrition, and carcass quality, have been less studied. In particular, further research is needed on mold species that contaminate hay post-flood, the effect of moldy hay ingestion on cattle, and the effects of heavy metal consumption on cattle and consumers. Little research is available regarding the effects of flooding on swine farms, likely due to the highly intensive nature of swine operations. In general, swine farms suffer from acute infectious diseases and biosecurity risks after flooding events. CONCLUSIONS: The potential ramifications of flooding on livestock farms differ among farm operations. Beef farms should prepare for long-term impacts on nutrition and reproduction, while swine farms should prepare for short-term impacts on biosecurity.

Effects of short-term moderate ZEN consumption on uterosacral ligament elasticity in pubertal gilts.

Zearalenone (ZEN) is a potent estrogenic toxin in swine, contributing to economic losses in herds via reproductive consequences such as pelvic organ prolapse (POP). To better understand the relationship between ZEN-consumption and reproductive symptoms, an animal feeding study with pubertal gilts was designed. The gilts were exposed to three different treatments: solvent-only feed for 21 days (n = 10), ZEN-spiked feed for 7 days followed by solvent-only feed for 14 days (n = 10), and ZEN-spiked feed for 21 days (n = 10). The gilts did not display any ZEN-related symptoms throughout any of the treatments. At the end of the trial the elastic properties of the USLs from participating gilts were evaluated along two loading directions: main direction (MD) and perpendicular direction (PD). The elastic properties included average stresses at 2% and 4% strains, and secant moduli. Overall the elastic properties of the USLs did not vary across treatment groups or between loading directions. In the MD, average stress increased from 32.96 ± 4.43 kPa at 2% strain to 63.21 ± 9.69 kPa at 4% strain, with a secant modulus of 1.52 ± 0.27 MPa. In the PD, average stress increased from 40.82 ± 4.22 kPa at 2% strain to 83.38 ± 9.17 kPa at 4% strain, with a secant modulus of 2.13 ± 0.31 MPa. Continued research into the relationship between ZEN consumption and reproductive symptoms such as POP is necessary in order to mitigate their deleterious effects in herds.

Removal of sows in Spanish breeding herds due to lameness: Incidence, related factors and reproductive performance of removed sows.

Lameness is a major reason for sow removal in breeding herds. Increased removal occurrences for lameness decrease reproductive efficiency and increase welfare concerns. Therefore, the objectives of this study were to estimate the incidence rate of removal due to lameness, and to investigate the longevity and reproductive performance of sows removed due to lameness. Poisson regression models were applied to a cohort dataset of 137,907 sows in 134 herds located in Spain. The Wilcoxon rank sum test was used to compare the performance of sows removed due to lameness and their controls in one-to-two matched case-control datasets. Removal due to lameness accounted for 4.3 % of all removed sows, and the incidence rate was 19.6 cases per 1000 sow-years (95 % confidence interval: 15.03, 25.51). The majority (70.4 %) of those removed were farrowed sows, whereas only 29.6 % were serviced sows. In farrowed sows, a higher incidence of removal due to lameness was associated with weeks 4-9 after farrowing, higher parity and winter farrowing (P < 0.01). The removal incidence was 24.7-33.1 times higher in weeks 4-9 after farrowing than during the first week after farrowing (P < 0.01). It was 1.3-1.6 times higher in parity 4-5 than in parity 1, and 1.3 times higher for winter farrowing than for summer farrowing (P < 0.01). In contrast, the factors associated with removal due to lameness with serviced sows were weeks 4-5 after service and being re-serviced (P < 0.01). The service sow removal incidence was 4.7 times higher in weeks 4-5 after servicing than during the first 2 weeks after servicing (P < 0.01). Also, it was 2.2 times higher in re-serviced sows than in first serviced sows (P < 0.01). However, removal in serviced sows was not associated with parity (P = 0.10) or service season (P = 0.39). In the case-control datasets, the sows removed due to lameness had higher weaning-to-first-mating interval (means: 6.5 vs. 5.8 days), fewer piglets born alive (11.7 vs. 12.5 piglets) and lower parity at removal (3.4 vs. 4.9; P < 0.01) than sows removed for other reasons or non-removed sows. However, there was no difference in gilt age at first service between the case and control groups (P = 0.29). We recommend identifying sows showing early signs of lameness and treating them with pain medication until removal. The best time for removal would be at weaning when non-productive sow days start.

Intrauterine growth restriction in piglets alters blood cell counts and impairs cytokine responses in peripheral mononuclear cells 24 days post-partum.

Large litter sizes have resulted in more piglets being exposed to intrauterine growth restriction (IUGR). Growth restriction during fetal life is linked with lower growth efficiency and increased susceptibility to infections in postnatal life and IUGR may associate with an altered innate immune system. We investigated the haematological, thromboelastography and plasma biochemical profiles of IUGR and normal piglets as well as cytokine responses in peripheral blood mononuclear cells stimulated with lipopolysaccharide (LPS) at 24 days of age. Piglets were classified at birth based on their head morphology as either IUGR or normal. The present study showed a modulation of the immune function of IUGR pigs, characterized by an increase in neutrophil percentage and fibrinogen levels but a decrease in CD4+ T-cells. A lower level of LPS-induced IL-1ß production was evident in IUGR pigs, suggesting immunological hypo-responsiveness. Furthermore, higher levels of reticulocytes, MCV and MCH and lower levels of erythrocytes in IUGR pigs suggest altered bone marrow hematopoiesis. All together, the results suggest a moderate suppression of the immune response of IUGR piglets, which may have implications for resistance to pathogen challenges in the post-weaning period. Serum metabolites and blood clotting profile did not differ between IUGR and normal piglets.

Time course determination of the effects of rapid and gradual cooling after acute hyperthermia on body temperature and intestinal integrity in pigs.

Rapid cooling after acute hyperthermia may cause a sustained increase in body temperature and exacerbate intestinal damage in pigs. Therefore, the study objective was to evaluate the temporal effects of rapid and gradual cooling on body temperature response and intestinal integrity after acute hyperthermia in pigs. In three repetitions, 54 pigs [83.3 ± 6.7 kg initial body weight (BW)], balanced by sex were exposed to thermoneutral conditions for 6 h (TN; n = 6 pigs/repetition; 21.1 ± 2.0°C), or heat stress conditions (HS; 39.3 ± 1.6°C) for 3 h, followed by a 3 h recovery period of gradual cooling [HSGC; n = 6 pigs/repetition; gradual decrease from HS to TN conditions] or rapid cooling [HSRC; n = 6 pigs/repetition; rapid TN exposure and cold water (4.0°C) dousing every 30 min for 1.5 h]. Feed was withheld throughout the entire 6 h period, but water was provided ad libitum. Gastrointestinal (TGI) and rectal (TR) temperatures were recorded every 15 min during the HS and recovery periods. Six pigs per repetition (n = 2/treatment) were euthanized and jejunal and ileal samples were collected for histology immediately after (d 0), 2 d after, and 4 d after the recovery period. Data were analyzed using PROC MIXED in SAS 9.4. Overall, rapid cooling reduced TR and TGI (P < 0.01; 0.95°C and 0.74°C, respectively) compared to gradual cooling. Jejunal villus height was reduced overall (P = 0.02; 14.01%) in HSGC compared to HSRC and TN pigs. Jejunal villus height-to-crypt depth ratio was reduced overall (P = 0.05; 16.76%) in HSGC compared to TN pigs. Ileal villus height was reduced overall (P < 0.01; 16.95%) in HSGC compared to HSRC and TN pigs. No other intestinal morphology differences were detected. In summary, HSRC did not cause a sustained increase in body temperature and did not negatively impact biomarkers of intestinal integrity in pigs.

The use of cells from ANPEP knockout pigs to evaluate the role of aminopeptidase N (APN) as a receptor for porcine deltacoronavirus (PDCoV).

The coronaviruses, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV) represent important sources of neonatal diarrhea on pig farms. The requirement for aminopeptidase N (APN) as a receptor for TGEV, but not for PEDV, is well established. In this study, the biological relevance of APN as a receptor for PDCoV was tested by using CRISPR/Cas9 to knockout the APN gene, ANPEP, in pigs. Porcine alveolar macrophages (PAMs) from ANPEP knockout (KO) pigs showed resistance to PDCoV infection. However, lung fibroblast-like cells, derived from the ANPEP KO PAM cultures, supported PDCoV infection to high levels. The results suggest that APN is a receptor for PDCoV in PAMs but is not necessary for infection of lung-derived fibroblast cells. The infection of the ANPEP KO pigs with PDCoV further confirmed that APN is dispensable as a receptor for PDCoV.

Hepatitis E Virus in Pigs from Slaughterhouses, United States, 2017-2019.

Hepatitis E virus (HEV) RNA was detected in 6.3% and HEV IgG in 40% of 5,033 serum samples from market-weight pigs at 25 slaughterhouses in 10 US states. The prevalent HEV genotype was zoonotic genotype 3, group 2. Blood of HEV-viremic pigs from slaughterhouses may contaminate pork supply chains.

Porcine Model of Infrarenal Abdominal Aortic Aneurysm.

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal abdominal aortic aneurysms (AAA) in swine. To achieve this, we use a combination of balloon angioplasty, elastase and collagenase, and a lysyl oxidase inhibitor, called ß-aminopropionitrile (BAPN), to create clinically significant infrarenal aortic aneurysms, analogous to human disease. Noncastrated male swine are fed BAPN for 7 days prior to surgery to achieve a steady state in the blood. A midline laparotomy is performed and the infrarenal aorta is circumferentially dissected. An initial measurement is recorded prior to aneurysm induction with a combination of balloon angioplasty, elastase (500 units)/collagenase (8000 units) perfusion, and topical elastase application. Swine are fed BAPN daily until terminal procedure on either postoperative day 7, 14, or 28, at which time the aneurysm is measured, and tissue procured. BAPN + surgery pigs are compared to pigs that underwent surgery alone. Swine treated with BAPN and surgery had a mean aortic dilation of 89.9% ± 47.4% at day 7, 105.4% ± 58.1% at day 14, and 113.5% ± 30.2% at day 28. Pigs treated with surgery alone had significantly smaller aneurysms compared to BAPN + surgery animals at day 28 (p < 0.0003). The BAPN + surgery group had macroscopic and immunohistochemical evidence of end stage aneurysmal disease. Clinically significant infrarenal AAA can be induced using balloon angioplasty, elastase/collagenase perfusion and topical application, supplemented with oral BAPN. This model creates large, clinically significant AAA with hallmarks of human disease. This has important implications for the elucidation of AAA pathogenesis and testing of novel therapies and devices for the treatment of AAA. Limitations of the model include variation in BAPN ingested by swine, quality of elastase perfusion, and cost of BAPN.

Kinetic Modelling of [68Ga]Ga-DOTA-Siglec-9 in Porcine Osteomyelitis and Soft Tissue Infections.

BACKGROUND: [68Ga]Ga-DOTA-Siglec-9 is a positron emission tomography (PET) radioligand for vascular adhesion protein 1 (VAP-1), a protein involved in leukocyte trafficking. The tracer facilitates the imaging of inflammation and infection. Here, we studied the pharmacokinetic modelling of [68Ga]Ga-DOTA-Siglec-9 in osteomyelitis and soft tissue infections in pigs. METHODS: Eight pigs with osteomyelitis and soft tissue infections in the right hind limb were dynamically PET scanned for 60 min along with arterial blood sampling. The fraction of radioactivity in the blood accounted for by the parent tracer was evaluated with radio-high-performance liquid chromatography. One- and two-tissue compartment models were used for pharmacokinetic evaluation. Post-mortem soft tissue samples from one pig were analysed with anti-VAP-1 immunofluorescence. In each analysis, the animal's non-infected left hind limb was used as a control. RESULTS: Tracer uptake was elevated in soft tissue infections but remained low in osteomyelitis. The kinetics of [68Ga]Ga-DOTA-Siglec-9 followed a reversible 2-tissue compartment model. The tracer metabolized quickly; however, taking this into account, produced more ambiguous results. Infected soft tissue samples showed endothelial cell surface expression of the Siglec-9 receptor VAP-1. CONCLUSION: The kinetics of [68Ga]Ga-DOTA-Siglec-9 uptake in porcine soft tissue infections are best described by the 2-tissue compartment model.

A porcine animal model to mimic the restart of enteral nutrition (refeeding-model).

With the aim towards establishing an animal model of total parenteral nutrition (TPN), 12 piglets aged 9 weeks (mean body weight 21 kg) were surgically provided with central venous catheters. Six piglets were nourished parenterally with the objective to reach a 14-d period of TPN; the other six piglets served as control and were fed normally. Only one animal from each group could be monitored over the whole period. Nine piglets were euthanised on d 13 and one on d 12. No animal showed fever or signs of septicaemia during the study. The levels of Ca, Mg, Na and P in the blood were within the normal range as were those for blood glucose and plasma creatinine. Symptoms of the TPN included: transient diarrhoea, occasional appearance of faecal blood and occasional absence of defecation. A reduced small intestine length and altered mucosal morphology and function were observed. One animal showed bile stasis at the end of the study. All TPN animals showed a remarkably high level of blood urea early in the morning. The intestinal symptoms observed may resemble the human situation during TPN. However, due to the fast growth rate, pigs aged 9 weeks have higher nutrient requirements per kg body weight. Consequently, the osmolality of the nutrient solution was necessarily high. Whether the significantly higher blood urea observed in the TPN group reflected a catabolic metabolism during the starving period at night-time could not be conclusively shown. Alternatively, it could reflect a slower growth rate and a resulting quantitative excess of amino acids (AA), or could have been the consequence of a suboptimal AA composition. A permanent infusion would be favourable in order not to overcharge the capacity for glucose uptake and amino acid metabolism during the infusion.

Acorn-related acquired pseudomelanosis in Calabrian black pigs.

BACKGROUND: Melanosis of lymph nodes in black pigs has generally been related to regression of congenital melanoma and, occasionally, to ingestion of acorns. The aim of this manuscript is to confirm the hypothesis of a possible acquired acorn-related pseudomelanosis in the Nero Calabrese pig, a swine breed belonging to the group of Italian native breeds and whose coverage area corresponds to the region of Calabria, southern Italy. This pig is characterized by slow-growing subjects, producing, however, high quality meat suitable for the production of sausages and fine hams. The study was carried out on 142 normally slaughtered pigs. All organs were examined. Lymph nodes and intestine (jejunum) were sampled. Histochemistry was performed on deparaffinized histological sections to identify the cell types involved and to characterize the pigment stored. To further confirm the pigmentation disorder, immunohistochemistry was carried out. Total phenolic substances were identified in acorns through the use of a biochemical reaction. RESULTS: Lymph node pigmentation appears directly related to acorn ingestion, with a higher incidence in the group which was 70% natural fed (acorn of Quercus virgiliana). Moreover, findings obtained revealed how different amounts of phenolic substrates present in Q. virgiliana and Q. ilex acorns can influence the incidence of such exogenous pigmentation. CONCLUSION: The findings obtained in this study confirm the acquired nature of the melanin-like pigmentation detected in lymph nodes from acorn-fed swine. Acquired pigmentation must be differentiated from true melanosis as well as from melanosis related to tumor regression of congenital melanoma. This thesaurismosis can be proposed as a marker of wellbeing and quality, confirming that the pigs have been bred and fed in natural conditions.

MicroRNAs in porcine uterus and serum are affected by zearalenone and represent a new target for mycotoxin biomarker discovery.

The mycotoxin zearalenone (ZEN) poses a risk to animal health because of its estrogenic effects. Diagnosis of ZEN-induced disorders remains challenging due to the lack of appropriate biomarkers. In this regard, circulating microRNAs (small non-coding RNAs) have remarkable potential, as they can serve as indicators for pathological processes in tissue. Thus, we combined untargeted and targeted transcriptomics approaches to investigate the effects of ZEN on the microRNA expression in porcine uterus, jejunum and serum, respectively. To this end, twenty-four piglets received uncontaminated feed (Control) or feed containing 0.17 mg/kg ZEN (ZEN low), 1.46 mg/kg ZEN (ZEN medium) and 4.58 mg/kg ZEN (ZEN high). After 28 days, the microRNA expression in the jejunum remained unaffected, while significant changes in the uterine microRNA profile were observed. Importantly, 14 microRNAs were commonly and dose-dependently affected in both the ZEN medium and ZEN high group, including microRNAs from the miR-503 cluster (i.e. ssc-miR-424-5p, ssc-miR-450a, ssc-miR-450b-5p, ssc-miR-450c-5p, ssc-miR-503 and ssc-miR-542-3p). Predicted target genes for those microRNAs are associated with regulation of gene expression and signal transduction (e.g. cell cycle). Although the effects in serum were less pronounced, receiver operating characteristic analysis revealed that several microRNA ratios were able to discriminate properly between non-exposed and ZEN-exposed pigs (e.g. ssc-miR-135a-5p/ssc-miR-432-5p, ssc-miR-542-3p/ssc-miR-493-3p). This work sheds new light on the molecular mechanisms of ZEN, and fosters biomarker discovery.