Sow-Offspring Diets Supplemented with Probiotics and Synbiotics Are Associated with Offspring's Growth Performance and Meat Quality.

Probiotics and synbiotics supplementation have been shown to play potential roles in animal production. The present study aimed to evaluate the effects of dietary probiotics and synbiotics supplementation to sows during gestation and lactation and to offspring pigs (sow-offspring) on offspring pigs' growth performance and meat quality. Sixty-four healthy Bama mini-pigs were selected and randomly allocated into four groups after mating: the control, antibiotics, probiotics, and synbiotics groups. After weaning, two offspring pigs per litter were selected, and four offspring pigs from two litters were merged into one pen. The offspring pigs were fed a basal diet and the same feed additive according to their corresponding sows, representing the control group (Con group), sow-offspring antibiotics group (S-OA group), sow-offspring probiotics group (S-OP group), and sow-offspring synbiotics group (S-OS group). Eight pigs per group were euthanized and sampled at 65, 95, and 125 d old for further analyses. Our findings showed that probiotics supplementation in sow-offspring diets promoted growth and feed intake of offspring pigs during 95-125 d old. Moreover, sow-offspring diets supplemented with probiotics and synbiotics altered meat quality (meat color, pH45min, pH24h, drip loss, cooking yield, and shear force), plasma UN and AMM levels, and gene expressions associated with muscle-fiber types (MyHCI, MyHCIIa, MyHCIIx, and MyHCIIb) and muscle growth and development (Myf5, Myf6, MyoD, and MyoG). This study provides a theoretical basis for the maternal-offspring integration regulation of meat quality by dietary probiotics and synbiotics supplementation.

Dynamic alteration in the gut microbiota and metabolome of Huanjiang mini-pigs during pregnancy.

BACKGROUND: Maternal gut microbiota and metabolites are associated with their offspring's health. Our previous study showed that maternal body fat percentage increased from days 45 to 110 of gestation in a Huanjiang mini-pig model. Thus, this study aimed to investigate the changes in gut microbiota composition and microbial metabolite profile of sows from days 45 to 110 of gestation. RESULTS: Twenty-four Huanjiang mini-pigs with average body weight were assigned for sample collection during early- (day 45 of pregnancy), mid- (day 75 of pregnancy), and late-pregnancy (day 110 of pregnancy). The results showed that the relative abundances of Clostridium_sensu_stricto_1, Romboutsia, Turicibacter, and Streptococcus in jejunal contents were higher at day 110 than those at day 45 or 75 of gestation. In the ileum, the relative abundance of Streptococcus was higher (P < 0.05) at day 110 of gestation, as well as the metabolism function of the jejunal and ileal microbiota. The ileal butyrate and acetate concentrations were higher at days 45 and 110 of gestation, respectively. In the colon, the concentrations of cadaverine and spermine were higher (P < 0.05) at days 45 and 110 of gestation, respectively. Metabolomic analyses demonstrated that the metabolic pathways, including D-glutamine and D-glutamate metabolism, phenylalanine/tyrosine/tryptophan biosynthesis, and alanine/aspartate/glutamate metabolism changed during gestation. CONCLUSION: Collectively, our results showed that gut microbiota composition and microbial metabolites changed dramatically from early to late pregnancy in a Huanjiang mini-pig model. These findings will provide new targets in formulating maternal nutritional interventions to alleviate the adverse effects during pregnancy on offspring health outcomes.

Betaine hydrochloride addition in Bama mini-pig's diets during gestation and lactation enhances immunity and alters intestine microbiota of suckling piglets.

BACKGROUND: Maternal nutrition during gestation and lactation is essential for offspring's health. The present study aimed to investigate the effects of betaine hydrochloride addition to sow diets during gestation and lactation on suckling piglet's immunity and intestine microbiota composition. Forty Bama mini-pigs were randomly allocated into two groups and fed a basal diet (control group) and a basal diet supplemented with 3.50 kg ton-1 betaine hydrochloride (betaine group) from day 3 after mating to day 21 of lactation. After 21 days of the delivery, 12 suckling piglets from each group with similar body weight were selected for sample collection. RESULTS: The results showed that maternal betaine hydrochloride addition decreased (P < 0.05) the plasma levels of interleukin (IL)-1ß, IL-2, IL-6, and tumor necrosis factor-α in suckling piglets. Furthermore, dietary betaine hydrochloride addition in sow diets increased (P < 0.05) the villus height (VH) and VH to crypt depth ratio in the jejunum and ileum of suckling piglets. In the piglets' intestinal microbiota community, the relative abundances of Roseburia (P < 0.05) and Clostridium (P = 0.059) were lower in the betaine group compared to those in the control group. Moreover, betaine hydrochloride addition in sow diets decreased the colonic tyramine (P = 0.091) and skatole (P = 0.070) concentrations in suckling piglets. CONCLUSION: Betaine hydrochloride addition in sow diets enhanced the intestinal morphology, improved immunity, and altered intestinal microbiota of suckling piglets. These findings indicated that betaine hydrochloride addition in sow diets during gestation and lactation will impact suckling piglets' health. © 2021 Society of Chemical Industry.

Dissection of the Trigeminal Ganglion of Nonrodent Species Used in Toxicology Studies.

The ganglion of the trigeminal (V cranial) nerve is generally sampled at necropsy in nonrodent toxicology studies only when somatic or autonomic peripheral nervous system toxicity is suspected. The ganglion is far more difficult to locate in nonrodents than in rats and mice, and suitable methods to dissect it have been described only for swine. The trigeminal nerve caudal to the ganglion passes through a canal, roofed by bone in dogs and rabbits and by a tough layer of dura mater in swine and nonhuman primates. The ganglion is partly or wholly obscured by overlying dura mater. Of the 3 intracranial branches of the nerve, the ophthalmic is delicate and the maxillary and mandibular have extremely short courses within the cranial cavity. Methods that are practical in routine toxicologic pathology for the dissection of the ganglion in nonrodent laboratory species are illustrated and relevant species differences in the anatomy of the intracranial part of the trigeminal nerve are highlighted.

[Methods for Creating Medium and Large Animal Models of Peripheral Atherosclerosis].

Atherosclerosis (AS) is the main pathological basis for the occurrence of many vascular diseases and has a strong association with metabolic syndrome. Peripheral arterial diseases caused by AS have high morbidities and mortalities but lack still effective treatment. AS animal models are highly valuable for research on peripheral arteriosclerotic diseases. While small and medium AS animal models based on high-fat feeding and balloon injuries have been successfully established,few literatures have described the creation of large animal models for AS treatment. This article elucidates the current methods for creating medium and large animal models of peripheral atherosclerotic disease,with an attempt to further promote the clinical translation of AS treatment research.

Perivascular adipose tissue dysfunction aggravates adventitial remodeling in obese mini pigs via NLRP3 inflammasome/IL-1 signaling pathway.

Perivascular adipose tissue (PVAT), a special type of adipose tissue, closely surrounds vascular adventitia and produces numerous bioactive substances to maintain vascular homeostasis. PVAT dysfunction has a crucial role in regulating vascular remodeling, but the exact mechanisms remain unclear. In this study, we investigated whether and how obesity-induced PVAT dysfunction affected adventitia remodeling in early vascular injury stages. Mini pigs were fed a high sugar and fat diet for 6 months to induce metabolic syndrome and obesity. In the mini pigs, left carotid vascular injury was then generated using balloon dilation. Compared with normal mini pigs, obese mini pigs displayed significantly enhanced vascular injury-induced adventitial responses, evidenced by adventitia fibroblast (AF) proliferation and differentiation, and adventitia fibrosis, as well as exacerbated PVAT dysfunction characterized by increased accumulation of resident macrophages, particularly the M1 pro-inflammatory phenotype, increased expression of leptin and decreased expression of adiponectin, and production of pro-inflammatory cytokines interleukin (IL)-1ß and IL-18. Primary AFs cultured in PVAT-conditioned medium from obese mini pigs also showed significantly increased proliferation and differentiation. We further revealed that activated nod-like receptor protein 3 (NLRP3) inflammasome and its downstream products, i.e., IL-1 family members such as IL-1ß and IL-18 were upregulated in the PVAT of obese mini pigs; PVAT dysfunction was also demonstrated in preadipocytes treated with palmitic acid. Finally, we showed that pretreatment with IL-1 receptor (IL-1R) antagonist or IL-1R knockdown blocked AF proliferation and differentiation in AFs cultured in PVAT-conditioned medium. These results demonstrate that obesity-induced PVAT dysfunction aggravates adventitial remodeling after early vascular injury with elevated AF proliferation and differentiation via activating the NLRP3/IL-1 signaling pathway.

Dietary nutrient levels alter the metabolism of arginine family amino acids in the conceptus of Huanjiang mini-pigs.

BACKGROUND: The arginine family amino acids (AFAAs) exert important roles in the metabolism, growth and development of the conceptus. However, to date, few studies have investigated the effects of maternal nutrient levels on the concentrations and metabolism of AFAAs in the conceptus. RESULTS: Compared to low nutrient diets, high nutrient diets increased (P < 0.05) the concentrations of citrulline and proline (Pro) in plasma; the concentrations of arginine, glutamine, Pro and ornithine (Orn) in the amniotic fluid; and the concentrations of all detected AFAAs in the allantoic fluid, which were most pronounced on day 45 of pregnancy. High nutrient diets upregulated (P < 0.05) mRNA expression of arginase I (Arg I), Pro oxidase and spermidine synthetase (SRM) in the fetal placenta, as well as Arg II, SRM and spermine synthetase (SMS) expression in the fetal liver (most pronounced on day 45 of pregnancy). The same effect was observed for mRNA expression of NO synthase and Orn aminotransferase (OAT), mainly on day 110 of pregnancy, and for mRNA expression of Arg I, Arg II, OAT, Orn decarboxylase and SMS throughout pregnancy. High nutrient diets upregulated (P < 0.05) mRNA expression of Y+ L-type amino acid transporter (LAT) and cationic amino acid transporter 1 (CAT1) in the fetal jejunum throughout pregnancy. Dietary treatments did not affect (P > 0.05) mRNA expression of Y+ LAT1, sodium-coupled neutral amino acid transporter 2 (SNAT2) and CAT1 in the fetal placenta, skeletal muscle and colon. CONCLUSION: High nutrient diets increased the concentration and transport of AFAAs in the mothers and conceptus, which likely improves growth and development of the conceptus. © 2018 Society of Chemical Industry.

The experimental study of shunt-decompression arterialized vein flap.

BACKGROUND: Arterialized vein flap is a kind of unphysiological flap. Unphysiological reconstruction of blood circulation leads to higher load than that supported by physiological flap and is the culprit of flap swelling, blood stasis, skin blistering, and necrosis after flap grafting. To resolve the multiple disadvantages of traditional flap grafting, by introducing the principles of fluid mechanics, shunt-decompression surgery is prepared to decline the circulation preload and improve the prognosis of arterialized vein flap grafting. METHODS: By introducing the principles of fluid mechanics, we established the model of shunt-decompression arterialized vein flap, which satisfied the common properties of general fluid that the interface pressure between object and fluid is reduced when the velocity of fluid is increased and vice versa-the effect of Bernoulli. Under this rule, we anastomose the arterialized vein to the branch of main artery of recipient region or make end-to-side anastomosis, which can maintain the blood flow of main artery, decrease the perfusion of flap, and preserve the decompressive effect of main artery to branches. From March, 2016 to September, 2016, we performed animal experiments on ten male bama mini pigs with average weight of 28 ± 2.35 kg. Superior epigastric artery of pig was used for feeding artery to arterialize the superficial epigastric veins. The total area of flap is 8 cm × 6 cm. End-to-side anastomosis and end-to-end anastomosis were established in experimental group and control group, respectively. Doppler speckle perfusion imaging apparatus was used to monitor the alterations of flap perfusion, blood flow of flap, tissue swelling and survival of flaps. RESULTS: The average flap perfusion (PU) at 1 week after surgery is 83.62 ± 3.14 in experimental group and 98.14 ± 6.54 in control group, respectively (P < 0.05), indicating the significant reduction of flap blood perfusion in experimental group as compared with control group. As to the survival of flaps, 7 flaps completely survived, 3 showed partial necrosis, and no one was found as complete necrosis in experimental group, while only 3 flaps survived, and 4 flaps and 3 flaps showed partial necrosis and complete necrosis in control group, respectively (P < 0.05). CONCLUSION: Based on the physiological features of arterialized vein flap and its problems in clinical application, we improved the anastomosis strategy of flap grafting and obtained excellent experimental outcomes, which provides an insight for the clinical application of arterialized vein flaps.

Metabolic adaptations to HFHS overfeeding: how whole body and tissues postprandial metabolic flexibility adapt in Yucatan mini-pigs.

PURPOSE: In the present study, we aimed to metabolically characterize the postprandial adaptations of the major tissues involved in energy, lipids and amino acids metabolisms in mini-pigs. METHOD: Mini-pigs were fed on high-fat-high-sucrose (HFHS) diet for 2 months and several tissues explored for metabolic analyses. Further, the urine metabolome was followed over the time to picture the metabolic adaptations occurring at the whole body level following overfeeding. RESULTS: After 2 months of HFHS consumption, mini-pigs displayed an obese phenotype characterized by high circulating insulin, triglycerides and cholesterol levels. At the tissue level, a general (muscle, adipose tissue, intestine) reduction in the capacity to phosphorylate glucose was observed. This was also supported by the enhanced hepatic gluconeogenesis potential, despite the concomitant normoglycaemia, suggesting that the high circulating insulin levels would be enough to maintain glucose homoeostasis. The HFHS feeding also resulted in a reduced capacity of two other pathways: the de novo lipogenesis, and the branched-chain amino acids transamination. Finally, the follow-up of the urine metabolome over the time allowed determining breaking points in the metabolic trajectory of the animals. CONCLUSIONS: Several features confirmed the pertinence of the animal model, including increased body weight, adiposity and porcine obesity index. At the metabolic level, we observed a perturbed glucose and amino acid metabolism, known to be related to the onset of the obesity. The urine metabolome analyses revealed several metabolic pathways potentially involved in the obesity onset, including TCA (citrate, pantothenic acid), amino acids catabolism (cysteine, threonine, leucine).

Dietary soy isoflavones differentially regulate expression of the lipid-metabolic genes in different white adipose tissues of the female Bama mini-pigs.

Soy isoflavones have been shown to affect lipid metabolism, however the underlying molecular mechanism(s) have not yet been fully understood. The present study, using female Bama mini-pig as a model, examined the effects of soy isoflavones on lipid metabolism and involved gene expression in different white adipose tissues. Female Bama Xiang mini-pigs of 35 days old were fed a basal diet (control, Con), or basal diet supplemented with increasing amounts of soy isoflavones (250, 500, or 1250 mg/kg diet) for 120 days. The results showed that soy isoflavones did not affect the body weight, but decreased the dorsal subcutaneous adipose tissue (DSA) mass and increased the mass of abdominal subcutaneous adipose tissue (ASA) and perirenal adipose tissue (PRA). Besides, soy isoflavones decreased the expression of lipogenic genes and increased the expression of lipolytic genes in DSA, while the opposite effects were observed in ASA and PRA. In addition, the expression of lipoprotein lipase was down regulated in DSA while up regulated in ASA and PRA by soy isoflavones. Moreover, the expression of estrogen receptors (ERs) was up regulated in DSA, and down regulated in ASA and PRA by soy isoflavones. Our results suggest that soy isoflavones affected the lipid metabolism in white adipose tissues of Bama mini-pigs in a site-specific manner, which might be mediated through PPARs and ERs regulated gene expression.

Pigs as Models to Test Cardiovascular Devices.

Pigs as laboratory animals are used in preclinical studies aimed at developing medical devices for cardiac surgery. The anatomy of the cardiovascular system of these animals has been well studied and acknowledged as suitable for use and the testing of new cardiovascular devices developed for humans. However, there are no morphometric characteristics of the aortic root and thoraco-abdominal part of porcine aorta. This can lead to difficulties in experimental surgery and even result in the death of experimental animals due to the mismatch in the size of the implantable devices. Thus, such information is essential to enhance the efficiency of surgical technologies used for eliminating aortic pathologies in their various sections. The purpose of our research is to study the anatomy of the aorta in mini pigs and to assess whether the size, age, and sex of the animals affect the size of the main structures in their aortas. In addition, we attempted to compare the results obtained by transesophageal echocardiography (TEE) and angiography. We studied 28 laboratory mini pigs, dividing them into three groups by body weight (40-70 kg, 71-90 kg, and 90 kg). We did not find any relationship between the external somatometric characteristics of the animals and the size of their aortas. Animals have individual anatomical variability in their cardiovascular systems, which means that they need to be examined in terms of preoperative planning by any available method-echocardiography, angiography, or multispiral computed tomography (CT).

Comparison of three methods for collecting interstitial fluid from subcutaneous tissue in mini pigs.

Interstitial fluid, owing to its similarity to blood components and higher sensitivity and specificity, finds widespread application in disease diagnosis and tumor marker detection. However, collecting interstitial fluid, particularly from the deep subcutaneous connective tissue, remains challenging.•This study aimed to compare three different collection methods - push-pull perfusion, multi-filament nylon thread implantation, and tissue centrifugation - for collecting interstitial fluid from the subcutaneous connective tissue layer of mini-pigs. High-performance ion chromatography was employed to analyze the conventional cation components in the samples and compare ion composition analysis between the different methods.•Results indicated that while the distribution of conventional cations in the interstitial fluid collected by the three methods was generally consistent, there were slight variations in the detection rates and concentrations of different ions. Hence, suitable collection methods should be selected based on the ions or collection sites of interest.

Dietary Protein Levels Modulate the Antioxidant Capacity during Different Growth Stages in Huanjiang Mini-Pigs.

Adequate crude protein (CP) levels in diets play potential roles in swine production. This study determined the impacts of different CP levels on the antioxidant capacity of pigs during different body weight (BW) stages. Three hundred and sixty Huanjiang mini-pigs were allocated to one of three independent experiments, including a 5−10 kg BW group, where CP levels included 14%, 16%, 18%, 20%, and 22%; a 10−20 kg BW group, where CP levels included 12%, 14%, 16%, 18%, and 20%; and a 20−30 kg BW group, where CP levels included 10%, 12%, 14%, 16%, and 18%. These independent experiments were conducted for 28, 28, and 26 days, respectively. Results showed that the 20% CP level increased (p < 0.05) the plasma CAT and GSH-Px activities and the GSH concentration of pigs than in the pigs supplemented with the 14−18% CP levels, and the 20% CP level up-regulated (p < 0.05) the ileal oxidative stress-related gene expression levels of pigs than in the pigs supplemented with the 14% CP level at the 5−10 kg BW. In addition, diets supplemented with 18% CP level increased (p < 0.05) the ileal GSH concentration of pigs than in the pigs supplemented with the 14% and 20% CP levels, and the 16−18% CP levels increased (p < 0.05) the jejunal SOD activity of pigs than in the pigs supplemented with the 14% CP level. At 10−20 kg BW, the 16% CP level presented the strongest jejunal and ileal antioxidant capacity, the 18% CP level had the lowest plasma concentrations of MDA and highest GSH, and the 14−16% CP levels increased the plasma CAT and SOD activities (p < 0.05). Moreover, the 16−20% CP levels up-regulated (p < 0.05) the oxidative stress-related gene expression levels. At 20−30 kg BW, diets supplemented with the 16% CP level increased the plasma CAT activity of pigs than in the pigs supplemented with the 12−14% CP levels, and the 14−16% CP levels decreased the MDA concentration compared with the 10% CP levels (p < 0.05). In conclusion, these findings indicate adequate CP levels of 20%, 16%, and 14% for Huanjiang mini-pigs at the 5−10, 10−20, and 20−30 kg BW stages, respectively.

Dietary Betaine Supplementation Enhances Colonic Barrier Function through the Nrf2/Keap1 and TLR4-NF-κB/MAPK Signaling Pathways and Alters Colonic Microbiota in Bama Mini-Pigs.

This study evaluated the effects of betaine supplementation in sows and/or their offspring's diets on the redox status, immune and inflammatory levels, colonic barrier function, and colonic microbial community of offspring piglets. Thirty-six Bama mini-sows on day 3 of gestation and their weaned offspring piglets (28 d of age) were randomly allocated to the following treatments: (1) sows and their weaned offspring fed the basal diet (control group, Con group); (2) sows fed the basal diet with 3.50 kg/t betaine, and their weaned offspring fed the basal diet (sows betaine group, SB group); (3) sows fed the basal diet with 3.50 kg/t betaine, and their weaned offspring fed the basal diet with 2.50 kg/t betaine (sow-offspring betaine group, S-OB group). Six offspring piglets from each group were selected to collect plasma and colon samples on d 30, 60, and 90 after weaning. Compared with the Con group, the plasma levels of IgA, IgM, GSH-Px, and SOD during d 30-90 after weaning, IFN-α, T-AOC, and GSH on d 30 and 60 after weaning were increased, while MDA during d 30-90 after weaning was decreased in the SB and S-OB groups (p < 0.05). In addition, the plasma levels of IFN-γ on d 60 and T-AOC on d 30 after weaning were higher in the S-OB group than those in the Con group (p < 0.05). In the colon, betaine supplementation increased plasma T-AOC, GSH, and SOD levels while decreasing MDA concentration (p < 0.05). Betaine supplementation improved the colonic protein abundances of ZO-1, occludin, and claudin in offspring and activated the Nrf2/Keap1 signaling pathway while inhibiting the TLR4-NF-κB/MAPK signaling pathway on d 90 after weaning. The 16S rRNA sequencing results showed that betaine supplementation altered colonic microbiota composition by increasing the relative abundances of Verrucomicrobia and Actinobacteria in the SB group while decreasing proinflammatory-associated microbiota abundances (Tenericutes, Prevotella, and Parabacteroides) (p < 0.05). Collectively, these findings suggest that dietary betaine supplementation in sows and/or their offspring could improve offspring piglets' redox status and immune and anti-inflammatory levels and enhance the colonic barrier function by activating Nrf2/Keap1 and inhibiting TLR4-NF-κB/MAPK signaling pathways.

Probiotics or synbiotics addition to sows' diets alters colonic microbiome composition and metabolome profiles of offspring pigs.

Little information exists about the effects of maternal probiotics and synbiotics addition on the gut microbiome and metabolome of offspring. The present study evaluated the effects of probiotics or synbiotics addition to sows' diets on colonic microbiota and their metabolites in offspring using 16S rRNA gene sequencing and metabolome strategy. A total of 64 pregnant Bama mini-pigs were randomly divided into control, antibiotic, probiotics, and synbiotics groups and fed the corresponding experimental diets during pregnancy and lactation. After weaning, two piglets per litter and eight piglets per group were selected and fed a basal diet. The ß-diversity analysis showed that the colonic microbiota of offspring had a clear distinction among the four groups at 65 days of age. Maternal probiotics addition increased the Actinobacteria abundance at 65 days of age and Tenericutes and Firmicutes abundances at 95 days of age of offspring compared with the other three groups, whereas maternal antibiotic addition increased Spirochaetes and Proteobacteria abundances at 95 days of age of offspring compared with the other three groups. Metabolomic analysis showed that colonic metabolites were different between the groups, regardless of the days of age. Furthermore, both PICRUSt2 and enrichment analysis of metabolic pathways showed that maternal probiotics and synbiotics addition affected metabolism of carbohydrate, amino acid, cofactors and vitamins in the colonic microbiota. Compared with the control group, the colonic concentration of indole decreased and skatole increased in the probiotics group, whereas indole increased and skatole decreased in the synbiotics group. Maternal probiotics addition increased the colonic concentrations of acetate and butyrate at 65 and 125 days of age, whereas probiotics and synbiotics addition decreased short-chain fatty acids concentrations at 95 days of age. In addition, the colonic concentrations of putrescine, cadaverine, 1,7-heptanediamine, and spermidine were increased in the antibiotic, probiotics, and synbiotics groups compared with the control group at 95 days of age. The correlation analysis showed that Gemmiger, Roseburia, and Faecalibacterium abundances were positively correlated with acetate, propionate, and butyrate concentrations; Gemmiger, Blautia, and Faecalibacterium were positively correlated with putrescine and spermidine; and Faecalibacterium, Blautia, Clostridium, and Streptococcus were positively correlated with (R)-3-hydroxybutyric acid. Collectively, these findings suggest that probiotics and synbiotics addition to sows' diets exerts effects on offspring pigs by altering gut microbiota composition and their metabolites. The potential beneficial effect on gut health is discussed.

Eliciting and Characterizing Porcine Vocalizations: When Pigs Fly.

BACKGROUND/OBJECTIVES: While voice-related therapeutic interventions are often researched preclinically in the porcine model, there are no well-established methods to induce porcine glottic phonation. Described approaches, such as training animals to phonate for positive reinforcement are time-consuming and plagued by inherent variability in the type of phonation produced and contamination of background noise. Thus, a reliable method of assessing glottic phonation in the porcine model is needed. METHODS: In this study, we have created a novel pulley-based apparatus with harness for "pig-lifting" with surrounding acoustic insulation and high-directional microphone with digital recorder for recording phonation. Praat and Matlab were used to analyze all porcine vocalizations for fundamental frequency (F0), intensity, duration of phonation and cepstral peak prominence (CPP). Glottic phonation was detected using F0 (≥2000 hz), duration (≥3 seconds) and researcher perceptual judgment. Partial-glottic phonations were also analyzed. Reliability between researcher judgment and acoustic measures for glottic phonation detection was high. RESULTS: Acoustic analysis demonstrated that glottic and partial-glottic phonation was consistently elicited, with no formal training of the minipigs required. Glottic vocalizations increased with multiple lifts. Glottic phonation continued to be elicited after multiple days but became less frequent. Glottic and partial-glottic phonations had similar CPP values over the 6 experimental days. CONCLUSION: Our cost-effective, reliable method of inducing and recording glottic phonation in the porcine model may provide a cost effective, preclinical tool in voice research.

Dietary Crude Protein Levels Alter Diarrhea Incidence, Immunity, and Intestinal Barrier Function of Huanjiang Mini-Pigs During Different Growth Stages.

Huanjiang mini-pig is an indigenous pig breed in China; however, the optimal dietary crude protein (CP) levels for this pig breed during different growth stages has not been standardized yet. This study investigated the effects of different CP levels on diarrhea incidence, immunity, and intestinal barrier function in pigs. A total of 360 Huanjiang mini-pigs were assigned to three independent trials and fed the following CP diets: 5-10 kg stage, 14, 16, 18, 20, and 22%; 10-20 kg stage, 12, 14, 16, 18, and 20% and 20-30 kg stage, 10, 12, 14, 16, and 18%. In the 5-10 kg stage, the 22%; diet increased the plasma IL-1ß, IL-6, IL-8, and TNF-α concentrations compared to the 14-20% diets and decreased IL-10 and TGF-ß; however, these results were fluctuated in the later stages, including the decrease of IL-1ß and IL-8 in the 20% group, TNF-α in the 18-20% groups, and the increase of IFN-γ in the 20% group at the 10-20 kg stage and the decrease of TNF-α in the 16% group at the 20-30 kg stage. The 20% diet increased the jejunal and ileal IL-10 concentration compared to the 14% diet at the 5-10 kg stage, as well as in the 16% diet compared to the 12% diet at the 10-20 kg stage. In addition, ileal IL-10 concentration was increased in the 16% diet compared to the 10, 12, and 18% diets at the 20-30 kg stage. Furthermore, the 18% diet at the 5-10 kg stage and the 16% diet at the 10-20 kg stage decreased jejunal IL-6 expression, whereas the 20% diet increased the TNF-α and IFN-γ at the 5-10 kg stage. The 20% diet increased the Claudin, Occludin, ZO-1, ZO-2, Mucin-1, and Mucin-20 expressions at the 5-10 kg stage, as well as TLR-2, TLR-4, and NF-κB in the 22 and 20% diets at the 5-10 and 10-20 kg stages, respectively. Collectively, these findings suggest optimal dietary CP levels of 16, 14, and 12% for Huanjiang mini-pigs during the 5-10, 10-20, and 20-30 kg growth stages, respectively; and provide the guiding significance of dietary CP levels for Huanjiang mini-pigs during different growth stages.

Probiotics and Synbiotics Addition to Bama Mini-Pigs' Diet Improve Carcass Traits and Meat Quality by Altering Plasma Metabolites and Related Gene Expression of Offspring.

This study evaluated the effects of maternal probiotics and synbiotics addition on several traits and parameters in offspring. A total of 64 Bama mini pigs were randomly allocated into the control (basal diet), antibiotic (50 g/t virginiamycin), probiotics (200 mL/day probiotics), or synbiotics (500 g/t xylo-oligosaccharides and 200 mL/day probiotics) group and fed with experimental diets during pregnancy and lactation. After weaning, two piglets per litter and eight piglets per group were selected and fed with a basal diet. Eight pigs per group were selected for analysis at 65, 95, and 125 days of age. The results showed that the addition of probiotics increased the average daily feed intake of the pigs during the 66- to 95-day-old periods and backfat thickness at 65 and 125 days of age, and that the addition of synbiotics increased backfat thickness and decreased muscle percentage and loin-eye area at 125 days of age. The addition of maternal probiotics increased the cooking yield and pH45min value at 65 and 95 days of age, respectively, the addition of synbiotics increased the meat color at 95 days of age, and the addition of probiotics and synbiotics decreased drip loss and shear force in 65- and 125-day-old pigs, respectively. However, maternal antibiotic addition increased shear force in 125-day-old pigs. Dietary probiotics and synbiotics addition in sows' diets increased several amino acids (AAs), including total AAs, histidine, methionine, asparagine, arginine, and leucine, and decreased glycine, proline, isoleucine, α-aminoadipic acid, α-amino-n-butyric acid, ß-alanine, and γ-amino-n-butyric acid in the plasma and longissimus thoracis (LT) muscle of offspring at different stages. In the LT muscle fatty acid (FA) analysis, saturated FA (including C16:0, C17:0, and C20:0) and C18:1n9t contents were lower, and C18:2n6c, C16:1, C20:1, and unsaturated FA contents were higher in the probiotics group. C10:0, C12:0, and C14:0 contents were higher in 65-day-old pigs, and C20:1 and C18:1n9t contents were lower in the synbiotics group in 95- and 125-day-old pigs, respectively. The plasma biochemical analysis revealed that the addition of maternal probiotics and synbiotics decreased plasma cholinesterase, urea nitrogen, and glucose levels in 95-day-old pigs, and that the addition of synbiotics increased plasma high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and total cholesterol concentrations in 65-day-old pigs and triglyceride concentration in 125-day-old pigs. The addition of maternal probiotics and synbiotics regulated muscle fiber type, myogenic regulation, and lipid metabolism-related gene expression of LT muscle in offspring. In conclusion, the addition of maternal probiotics and synbiotics improved the piglet feed intake and altered the meat quality parameters, plasma metabolites, and gene expression related to meat quality.

New Therapy for Spinal Cord Injury: Autologous Genetically-Enriched Leucoconcentrate Integrated with Epidural Electrical Stimulation.

The contemporary strategy for spinal cord injury (SCI) therapy aims to combine multiple approaches to control pathogenic mechanisms of neurodegeneration and stimulate neuroregeneration. In this study, a novel regenerative approach using an autologous leucoconcentrate enriched with transgenes encoding vascular endothelial growth factor (VEGF), glial cell line-derived neurotrophic factor (GDNF), and neural cell adhesion molecule (NCAM) combined with supra- and sub-lesional epidural electrical stimulation (EES) was tested on mini-pigs similar in morpho-physiological scale to humans. The complex analysis of the spinal cord recovery after a moderate contusion injury in treated mini-pigs compared to control animals revealed: better performance in behavioural and joint kinematics, restoration of electromyography characteristics, and improvement in selected immunohistology features related to cell survivability, synaptic protein expression, and glial reorganization above and below the injury. These results for the first time demonstrate the positive effect of intravenous infusion of autologous genetically-enriched leucoconcentrate producing recombinant molecules stimulating neuroregeneration combined with neuromodulation by translesional multisite EES on the restoration of the post-traumatic spinal cord in mini-pigs and suggest the high translational potential of this novel regenerative therapy for SCI patients.

A miniature pig model of pharmacological tolerance to long-term sedation with the intravenous benzodiazepines; midazolam and remimazolam.

As a new and ultra fast-acting IV benzodiazepine, pharmacological tolerance may be anticipated during long-term treatment with remimazolam e.g. in intensive care. In this context, tolerance is particularly relevant for withdrawal syndrome. However, apart from primates, existing models of sedative tolerance are unsuitable for remimazolam due to its excessive metabolic clearance (i.e. in rodents) or paradoxical responses (in dogs). Pigs are a well-established model species, especially for in-vivo drug safety studies, and appear a well suited as model for evaluation of remimazolam. In a series of experiments from dose-range-finding bolus and infusion studies through to 28-day continuous level sedation, we established a viable model of intravenous benzodiazepine sedation in NIBS micropigs to compare tolerance development during 28 days sedation with either midazolam or remimazolam. Dose increases after 28 days were lower for remimazolam (0 to 3-fold) than for midazolam (2 to 4-fold) and recovery times were approximately 40% faster for remimazolam vs midazolam. Tolerance to remimazolam is therefore likely in long-term human sedation and may be less than that seen for midazolam.