Intact Pigs Research Articles

Determining underlying influences of data variability in the novel object recognition paradigm as used with young pigs.

The novel object recognition (NOR) paradigm is a cognitive test that has been used with many species to detect differences in ability. Various iterations of the paradigm have been implemented, making it difficult to compare results both within and across species. Interpretations of the results are equally diverse, threatening the integrity of the paradigm. These inconsistencies have prompted a deeper dive into the variability of the resultant data. For the purposes of this meta-analysis, data originated from 12 studies involving 367 pigs that were subjected to the same NOR paradigm beginning between postnatal days 21 and 24. The main cognitive measure from the NOR paradigm is recognition index (RI), which was the focus of most of the analyses in this meta-analysis. RI was chosen as the main outcome as it determines a pig's preference for novelty, an innate behavior of cognitively intact pigs. A histogram of RI values (range 0 to 1) showed a bimodal distribution skewed to the right, suggesting that the interpretation of positive performance on the task may need to be stricter. Correlational analyses proved that the number of investigations and investigation time with both the novel and familiar objects were the strongest predictors of resultant RI values. Objective data inclusion criteria were then considered to eliminate non-compliant pigs. Results indicated that requiring at least 5 s of investigation over a minimum of 3 investigations with the novel object reduced overall variability for RI with a concomitant increase in the mean. Further analyses showed that pigs preferred to spend more time with and interact more with the novel object across the entire testing trial, especially in the first minute. Together, these findings suggest that future interpretations of NOR should consider applying stricter statistical analyses as well as additional data processing, such as binning, with emphasis on novel object and familiar object investigation. Overall, modifications to the existing iterations of the NOR paradigm are necessary to improve paradigm reliability.

23 The effects of dietary net energy level on grow-finish performance and carcass characteristics of intact male market pigs managed with Improvest

Abstract Improvest is a gonadotropin releasing factor (GnRF) analogue-diphtheria toxoid conjugate product approved for temporary suppression of testicular function and reduction of boar taint in intact male pigs (Zoetis Canada Inc.). Objectives for this study were to determine the effects of dietary net energy (NE) level during the grow-finish period on live performance and carcass characteristics of intact male pigs managed with Improvest/immunological castration (IC) when compared with physically castrated (PC) male pigs. The 101-d study began when 1,008 pigs (504 intact male pigs and 504 PC male pigs; 10 wk old) were allocated by body weight to 48 pens with 21 intact males or 21 PC males per pen. Three dietary NE treatments were fed to pigs using five dietary phases to provide an average of 2,212 kcal/kg (Low NE), 2,337 kcal/kg (Medium NE), or 2,462 kcal/kg (High NE). Dietary programs were formulated according to standardized ileal digestible lysine requirements of IC males or PC males. The experiment was designed and analyzed as a 2 × 3 factorial with main effects of Improvest management and NE. For the overall study period, there were no significant interactions between Improvest management and NE (P ≥ 0.19) for average daily feed intake (ADFI), average daily gain (ADG), or gain to feed ratio (G:F; Table). There were also no significant interactions between Improvest management and NE (P ≥ 0.06) for carcass characteristics. Immunologically castrated male pigs consumed less feed (5.3% less ADFI; P < 0.01), grew faster (5.1% greater ADG; P < 0.01), and were more efficient (11.2% greater G:F; P < 0.01) compared with PC male pigs. Hot carcass weight (HCW) did not differ (P = 0.16) between IC males and PC males (attributed to 1.6 percentage unit less dressing percentage for IC males; P < 0.01); however, IC males were leaner (0.9 mm less backfat and 0.65% greater predicted lean yield; P < 0.01) compared with PC males. For the overall study period, pigs fed Low NE and Medium NE diets consumed more feed than pigs fed High NE diets, and pigs fed Low NE diets grew slower than pigs fed Medium NE and High NE diets. This resulted in pigs fed Low NE diets having 4.4% less G:F than pigs fed Medium NE and 8.6% less G:F than pigs fed High NE diets (P < 0.01). Pigs fed Low NE had 3.0 kg lighter (P < 0.01) HCW compared with Medium NE, while High NE had intermediate HCW that did not differ from the other two treatments. Overall, typical Improvest response levels were sustained for each of the NE treatments in this study; however, consideration should be provided to the known production impacts of low NE diets when male market pigs are managed with Improvest.

Effect of SNPs in the Promoter Region on the Expression of Cytochrome P450 2E1 (CYP2E1) in Pig Liver.

Boar taint, an unfavorable odor in the meat of intact male pigs, is caused primarily by the accumulation of two compounds: androstenone and skatole. This multifactorial trait is regulated by numerous dietary, management and genetic factors. At the mechanistic level, there are many genes known to be involved in boar taint metabolism. Cytochrome P450 2E1 (CYP2E1) impacts boar taint through the phase I metabolism of skatole. The aim of this study was to identify single-nucleotide polymorphisms (SNPs) within the CYP2E1 gene promoter and explore their relationship with the expression of CYP2E1 mRNA and protein. Sequencing of the promoter region using pools of genomic DNA identified seven promoter region SNPs at -159, -586, -1693, -1806, -2322, -2369 and -2514 bp upstream of the ATG start site. Genomic DNA was obtained from 65 boars from the three major swine breeds: Duroc, Landrace and Yorkshire, and individual animals were genotyped for the identified SNPs. RNA was isolated from liver tissue and quantitative PCR was performed to measure CYP2E1 gene expression, while levels of CYP2E1 protein in liver were measured by Western blotting. Significant within-breed variation in CYP2E1 protein and mRNA expression was observed, indicating significant differences in gene expression among individuals. However, levels of CYP2E1 mRNA and protein were not significantly correlated. Two SNPs within the promoter were significantly associated with CYP2E1 mRNA expression, but not with protein expression. This study provides evidence of additional mutations affecting the gene expression of CYP2E1 and suggests that factors that affect the differences in translation of CYP2E1 mRNA may also be important in affecting skatole metabolism.

Differential Regulation of Male-Hormones-Related Enhancers Revealed by Chromatin Accessibility and Transcriptional Profiles in Pig Liver.

Surgical castration can effectively avoid boar taint and improve pork quality by removing the synthesis of androstenone in the testis, thereby reducing its deposition in adipose tissue. The expression of genes involved in testis-derived hormone metabolism was altered following surgical castration, but the upstream regulatory factors and underlying mechanism remain unclear. In this study, we systematically profiled chromatin accessibility and transcriptional dynamics in liver tissue of castrated and intact full-sibling Yorkshire pigs. First, we identified 897 differentially expressed genes and 6864 differential accessible regions (DARs) using RNA- and ATAC-seq. By integrating the RNA- and ATAC-seq results, 227 genes were identified, and a significant positive correlation was revealed between differential gene expression and the ATAC-seq signal. We constructed a transcription factor regulatory network after motif analysis of DARs and identified a candidate transcription factor (TF) SP1 that targeted the HSD3B1 gene, which was responsible for the metabolism of androstenone. Subsequently, we annotated DARs by incorporating H3K27ac ChIP-seq data, marking 2234 typical enhancers and 245 super enhancers involved in the regulation of all testis-derived hormones. Among these, four typical enhancers associated with HSD3B1 were identified. Furthermore, an in-depth investigation was conducted on the androstenone-related enhancers, and an androstenone-related mutation was identified in a newfound candidatetypical enhancer (andEN) with dual-luciferase assays. These findings provide further insights into how enhancers function as links between phenotypic and non-coding area variations. The discovery of upstream TF and enhancers of HSD3B1 contributes to understanding the regulatory networks of androstenone metabolism and provides an important foundation for improving pork quality.

The effects of dietary net energy on grow-finish performance and carcass characteristics of male market pigs managed with immunological castration (Improvest).

The objective was to determine the effects of dietary net energy (NE) during the grow-finish period on live performance and carcass characteristics of intact male pigs managed with immunological castration (Improvest) compared with physically castrated (PC) male pigs. The 101-d study began when 1,008 pigs (504 intact male pigs and 504 PC male pigs; 10wk old) were allocated by weight to 48 pens with 21 intact males or 21 PC males per pen. Three dietary NE treatments were fed to pigs using five dietary phases (dietary programs were formulated according to standardized ileal digestible lysine requirements of Improvest males or PC males) to provide an average of 2,212 kcal/kg (low NE), 2,337 kcal/kg (medium NE), or 2,462 kcal/kg (high NE). The experiment was designed and analyzed as a 2 × 3 factorial with main effects of Improvest management and NE. For the overall study period, there were no significant interactions between Improvest management and NE (P ≥ 0.19) for average daily feed intake (ADFI), average daily gain (ADG), or gain:feed (G:F). There were also no significant interactions between Improvest management and NE (P ≥ 0.06) for carcass characteristics. Improvest males consumed less feed (5.3% lower ADFI; P < 0.01), grew faster (5.1% greater ADG; P < 0.01), and were more efficient (11.2% greater G:F; P < 0.01) compared with PC males. Hot carcass weight (HCW) did not differ (P = 0.16) between Improvest males and PC males (attributed to 1.6 percentage unit lower dressing percentage for Improvest males; P < 0.01); however, Improvest males were leaner (0.9mm less backfat and 0.65% greater predicted lean yield; P < 0.01) compared with PC males. For the overall study period, pigs fed low NE and medium NE diets consumed 7.5% and 4.6% more feed (P < 0.01) than pigs fed high NE diets, respectively, and pigs fed low NE diets grew 1.7% slower (P < 0.02) than pigs fed medium NE and high NE diets. This resulted in pigs fed low NE diets having 4.4% lower G:F compared with pigs fed medium NE and 8.6% lower G:F compared with pigs fed high NE diets (P < 0.01). Pigs fed low NE had 3.0kg lighter (P < 0.01) HCW compared with medium NE, while high NE had intermediate HCW that did not differ from the other two treatments. Overall, typical Improvest response levels for live performance and carcass characteristics when compared with PC males were achieved for each of the NE treatments evaluated in this study.

Results of ligation the internal iliac and ovarian arteries in the experiment

Objective: to study in an experiment on animals pathomorphological changes in the tissues of the uterus, appendages and blood vessels after ligation of the internal iliac and ovarian arteries.Materials and methods: An experiment was conducted with 6 mature pigs. Group I (n=3) consisted of intact pigs without surgery, group II (n=3) included animals with ligation of the internal iliac and ovarian arteries on both sides. 12 months after the operation, slaughter was performed and the organo-complex of the uterus with appendages and iliac vessels was taken for electron microscopic pathomorphological examination.Results: comparative results of the study of the uterus, appendages found that the normal structure of the mature uterus was preserved, there were no changes in the structure of the fallopian tubes, tissues and vessels of the ovaries. All changes were found in the area of ligation of the ovarian and internal iliac arteries, although the arteries retained their lumen, but were deformed and surrounded by dense fibrous connective tissue. Conclusion: ligation of the ovarian and internal iliac arteries with absorbable suture material causes only local changes. The sufficiency of collateral blood flow is confirmed by the absence of atrophy of the uterus, fallopian tubes and ovaries, as well as organ deformities.

Influence of Sex and a High-Fiber Diet on the Gut Microbiome of Alentejano Pigs Raised to Heavy Weights.

This study investigates the influence of sex and a dietary transition on the gut microbiota of a local Portuguese pig breed. Three groups of male Alentejano pigs (n = 10 each) were raised between ~40 and 160 kg LW. Group C included pigs that were surgically castrated, while the I group included intact ones; both were fed with commercial diets. The third group, IExp, included intact pigs that were fed commercial diets until ~130 kg, then replaced by an experimental diet based on legumes and agro-industrial by-products between ~130 and 160 kg. Fecal samples were collected two weeks before slaughter. The total DNA was extracted and used for 16S metabarcoding on a MiSeq® System. The dietary transition from a commercial diet to the experimental diet substantially increased and shifted the diversity observed. Complex carbohydrate fermenting bacteria, such as Ruminococcus spp. and Sphaerochaeta spp., were significantly more abundant in IExp (q < 0.05). On the other hand, castrated pigs presented a significantly lower abundance of the potential probiotic, Roseburia spp. and Lachnospiraceae NK4A136 group (q < 0.01), bacteria commonly associated with better gut health and lower body fat composition. Understanding the role of gut microbiota is paramount to ensure a low skatole deposition and consumers' acceptance of pork products from non-castrated male pigs.

Influence of Sex on Meat and Fat Quality from Heavy Alentejano Pigs Finished Outdoors on Commercial and High Fiber Diets.

This work aimed to examine the effects of sex on meat and fat quality traits from thirty Portuguese Alentejano (AL) pigs reared in outdoor conditions. These pigs were divided into three groups and fed ad libitum. From ~40 to 130 kg LW, castrated (C group) and intact animals (I and IExp) consumed commercial diets. Until slaughter (~160 kg), C and I pigs remained on commercial diets, and IExp changed to a more sustainable experimental diet with locally produced pulses and byproducts. Samples were collected from the Longissimus lumborum (LL), Psoas major (PM), and dorsal subcutaneous fat (DSF). At ~160 kg, the PM muscle of intact pigs presented lower intramuscular fat content than that of C pigs, while total collagen was higher. Additionally, PM myoglobin was lower and lightness (L*) was higher in intact pigs. Regarding DSF, moisture and total protein contents were higher and total lipids were lower in intact than in castrated pigs, while color parameters were not significantly different. Finally, antioxidant capacity measured in the LL muscle showed an overall lower value in intact pigs. However, lipid oxidation values were not significantly different between the experimental groups and only increased with storage time. Outdoor-reared intact AL pigs produced leaner and less saturated pork and fat compared to castrated ones. Despite the lower antioxidant activity observed in the LL muscles of intact pigs, the lipid oxidative stability of cooked meat was not different among the experimental groups.

Transcriptomic Profiling of Subcutaneous Backfat in Castrated and Intact Alentejano Pigs Finished Outdoors with Commercial and Fiber-Rich Diets.

In this work, we studied the backfat transcriptome of surgically castrated (C), intact (I) and intact fed an experimental diet (IE) outdoor-reared male Alentejano (AL) pigs. The experimental diet was a high-fiber diet with locally produced legumes and by-products associated with a boar taint reduction effect. At slaughter (~160 kg), backfat samples were collected for total RNA sequencing. Intact pigs presented leaner carcasses, more total collagen, and more unsaturated intramuscular fat content than C animals. A total of 2726 differentially expressed genes (DEGs, |log2 FC|> 0.58, q < 0.05) were identified between C and I with overexpressed genes related to muscular activity (MYH1, ACTA1) or collagen metabolism (COL1A1, COL1A2) in I pigs. Between C and IE, 1639 DEGs of genes involved in lipidic metabolism (LEP, ME1, FABP4, ELOVL6) were overexpressed in C. Finally, only 28 DEGs were determined between I and IE. Clustering results indicated a drastic influence of the testis in the transcriptome of subcutaneous fat of AL pigs, while the diet had a marginal effect. Diet can reduce stress by increasing satiety in animals, and could have induced an increase of skatole degradation due to the higher expression of the CYP2A19 gene in the IE group.

Effect of mixing at weaning and nutrient density of the weaner diet on growth performance and welfare of pigs to slaughter

BackgroundMixing pigs at weaning can compromise pig welfare and growth. Therefore, grouping littermates together may allow a diet nutrient and energy density reduction during the nursery period to reduce feed cost without affecting slaughter weight. This study investigated the combined effect of mixing and reducing dietary energy and nutrient density on growth performance, body lesions (BL), and behaviour in pigs from weaning to slaughter.ResultsForty-eight litters [554 pigs, 11–12 pigs/litter; Danish Duroc × (Large White × Landrace)] were included in the trial. At 28 days of age, pigs were weaned and housed in nursery rooms in litter groups (INTACT, n = 24) or mixed with other litters and grouped by weight to reduce within-pen pig weight variation (MIXED, n = 24). A dietary regimen meeting pigs’ nutritional requirements (CON) and a low-density dietary regimen (LOW; -10% energy and protein) completed a 2 × 2 factorial arrangement (Mixing x Diet, n = 12). On day 74 of age, pigs moved to the grower-finisher accommodation without further mixing and all pigs received the CON dietary regimen. Mixing increased FCR by 4.0% during the nursery period (p = 0.003). Nursery pigs fed LOW experienced a growth retardation which was maintained until slaughter (-2.6 kg slaughter weight; p = 0.025). Initial differences in the coefficient of variation (CV) between MIXED (10.4%) and INTACT (17.6%; p < 0.001) pigs were reduced in CON pens but not in LOW pens (interaction p = 0.025) at the end of the nursery period. MIXED pigs had more fights and BL (p < 0.001) at weaning and showed more aggression (p = 0.003) after being moved to the grower-finisher rooms. At the end of the nursery period, MIXED pigs fed LOW showed the highest number of aggressive behaviours around the feeder (interaction; p = 0.003) and pigs fed LOW showed more damaging behaviour (p < 0.001).ConclusionsMixing animals at weaning had limited impact on growth performance but impaired welfare which was aggravated by energy and nutrient reduction in the nursery diet. Decreasing dietary nutrient density in the nursery stage retarded growth, which could not be compensated for during the growing-finishing period.

Outdoor Finishing of Intact Male Portuguese Alentejano Pigs on a Sustainable High-Fiber Diet: Impacts on Blood, Growth, Carcass, Meat Quality and Boar Taint Compounds

This trial evaluated the effect of sex on the blood, growth, carcass, meat quality, and boar taint compounds in male Alentejano (AL) pigs (n = 30). From ~40 to 130 kg LW, castrated (C) and intact pigs (I and IExp groups) were fed commercial diets ad libitum. Between ~130 and 160 kg (slaughter), C and I pigs continued on commercial diets, while IExp were fed an experimental diet containing locally produced pulses and by-products aimed at reducing boar taint. At ~160 kg, blood urea levels were higher in IExp than C pigs, triacylglycerols were lower in both intact groups, and cortisol was lower in IExp. IExp pigs exhibited faster growth, improved feed conversion ratio, carcass higher commercial yield and leaner meat than C pigs. The loin intramuscular fat in intact pigs was lower, less saturated and more polyunsaturated, while total collagen was higher. Fat androstenone content was higher in intact pigs and skatole content was similar across treatments, although they were below threshold values for consumer detection. Finally, although boar taint compounds were low in intact AL pigs raised outdoors, adding pulses and by-products to the experimental diet did not result in a reduction in fat skatole content compared to pigs fed the commercial diet.

Some cardiopulmonary effects of capnoperitoneum in anesthetized guinea pigs (Cavia porcellus): spontaneous ventilation versus intubation and mechanical ventilation

ObjectiveTo compare cardiopulmonary variables and blood gas analytes in guinea pigs (Cavia porcellus) during anesthesia with and without abdominal carbon dioxide (CO2) insufflation at intra-abdominal pressures (IAPs) 4 and 6 mmHg, with and without endotracheal intubation. Study designProspective experimental trial. AnimalsA total of six intact female Hartley guinea pigs. MethodsA crossover study with sequence randomization for IAP and intubation status was used. The animals were sedated with intramuscular midazolam (1.5 mg kg–1) and buprenorphine (0.2 mg kg–1) and anesthetized with isoflurane, and an abdominal catheter was inserted for CO2 insufflation. Animals with endotracheal intubation were mechanically ventilated and animals maintained using a facemask breathed spontaneously. After 15 minutes of insufflation, the following variables were obtained at each IAP: pulse rate, respiratory rate, rectal temperature, oxygen saturation, end-tidal CO2 (intubated only), peak inspiratory pressure (intubated only), noninvasive blood pressure and blood gas and electrolyte values, with a rest period of 5 minutes between consecutive IAPs. After 4 weeks, the procedure was repeated with the guinea pigs assigned the opposite intubation status. ResultsIntubated guinea pigs had significantly higher pH and lower partial pressure of CO2 in cranial vena cava blood (PvCO2) than nonintubated guinea pigs. An IAP of 6 mmHg resulted in a significantly higher PvCO2 (65.9 ± 19.0 mmHg; 8.8 ± 2.5 kPa) than at 0 (53.2 ± 17.2 mmHg; 7.1 ± 2.3 kPa) and 4 mmHg (52.6 ± 10.8 mmHg; 7.01 ± 1.4 kPa), mean ± standard deviation, with intubated and nonintubated animals combined. Conclusions and clinical relevanceAlthough the oral anatomy of guinea pigs makes endotracheal intubation difficult, capnoperitoneum during anesthesia induces marked hypercapnia in the absence of mechanical ventilation. An IAP of 4 mmHg should be further evaluated for laparoscopic procedures in guinea pigs because hypercapnia may be less severe than with 6 mmHg.

Assessment of laparoscopic working space in guinea pigs (Cavia porcellus) undergoing carbon dioxide insufflation at different intra-abdominal pressures.

To evaluate pneumoperitoneal volumes (laparoscopic working space) in guinea pigs (Cavia porcellus) undergoing pneumoperitoneum via carbon dioxide insufflation at different intra-abdominal pressures (IAPs) (4, 6, and 8 mm Hg) and recumbencies (dorsal, right lateral, and left lateral). Six 3- to 4-month-old sexually intact female Hartley guinea pigs. Guinea pigs were anesthetized, intubated, and had an abdominal insufflation catheter placed. A baseline abdominal CT scan was performed. Guinea pigs underwent insufflation, with each IAP given in a random order for 10 to 15 minutes with a washout period of 5 minutes between pressures. Abdominal CT scans were acquired at each IAP and at each recumbency. Pneumoperitoneal volumes were calculated using software. Increases in IAP increased working space significantly (P < .001). The 6- and 8-mm Hg pressures increased working space from 4 mm Hg by 7.3% and 19.8%, respectively. Recumbent positioning (P = .60) and body weight (P = .73) did not affect working space. Order of IAP had a significant (P = .006) effect on working space. One of the guinea pigs experienced oxygen desaturation and bradycardia at 6- and 8-mm Hg IAP. Although an increased working space occurred at 6 and 8 mm Hg compared to 4 mm Hg, further research is needed concerning the cardiovascular effects of pneumoperitoneum in guinea pigs to determine whether those higher IAPs are safe in this species. An IAP of 6 mm Hg can be considered for laparoscopic cannula placement, followed by a lower IAP for laparoscopic procedures.

Effect of pneumoperitoneum on gastrointestinal motility, pain behaviors, and stress biomarkers in guinea pigs (Cavia porcellus).

To compare stress markers, gastrointestinal motility, and behavioral indicators of pain between guinea pigs undergoing pneumoperitoneum with carbon dioxide (CO2) and control guinea pigs. Fourteen 4- to 5-month-old intact female Hartley guinea pigs. Guinea pigs were randomized to receive insufflation or serve as controls (anesthesia and abdominal catheter placement without insufflation), with 7 animals/group. Insufflated animals underwent 6 mm Hg of CO2 pneumoperitoneum for 30 minutes. Afterward, results for vital signs, blood glucose, fecal cortisol, appetite, fecal output, and behaviors (via video recording) were compared between the 2 groups. There was no difference between groups and over time for body temperature, heart rate, fecal output in grams, pellets consumed, blood glucose, and fecal cortisol. Guinea pigs that underwent insufflation had significantly more fecal pellets at 36 hours after the procedure. Several behaviors were expressed similarly between groups and over time, such as body turns, incomplete movement, rearing, lying down, drinking, and hiding. Coprophagy occurred less often in the insufflated versus noninsufflated group at 12 h postprocedure but was similar between groups at other time points. At 60 hours after the procedure, insufflated animals spent less time squinting compared to noninsufflated animals. Other behaviors were differentially expressed over time but not between treatments. Overall, there were no major differences in appetite, stress markers, and behaviors between insufflated and control guinea pigs. CO2 insufflation did not appear to cause undue pain or stress in guinea pigs and may be a reasonable technique to use during laparoscopy.

Estrogen receptor alpha expression in fattening pig’s testes by the timing of the first injection for immunocastration

The purpose of this study was to examine the expression of estrogen receptor alpha (ERα) in the testicular tissues of male fattening pigs injected with the first gonadotropin-releasing hormone (GnRH) vaccine (Improvac™, Zoetis, Thailand) for immunocastration 6 weeks earlier than the standard protocol. All pigs (n=24) were divided into three groups on the criterion of immunocastration protocol: A (n=8) was injected with GnRH vaccine at 15 and 19 weeks old, B (n=8) received GnRH vaccine at 9 and 19 weeks old, and C (n=8) remained untreated. Expression of ERα was investigated using an immunohistochemistry, appraised by an image analysis application (3DHISTECH, Budapest, Hungary), and reported as a H-score. The results revealed that testicular histoarchitecture of the immunocastrated pigs was less developed than that of the intact pigs. ERα was localized both in the seminiferous tubules and interstitial areas of all groups. ERα H-score of the C pigs was lowest (5.49±4.17) among groups and significantly lower than that of both A and B groups (P&lt;0.05). However, the H-score of ERα between A and B groups was not different from each other (21.41±12.61 vs 23.91±11.47, P&gt;0.05). In summary, the first injection of GnRH vaccine either at 9 or 15 weeks of age contributed to a similar result of testicular ERα expression.

Sus_circPAPPA2 Regulates Fat Deposition in Castrated Pigs through the miR-2366/GK Pathway.

CircRNAs play an important role in fat deposition, and testosterone-deficient boars exhibit significantly increased fat deposition; however, the mechanism by which testosterone regulates fat deposition through circRNAs remains unclear. In this study, circRNA-seq of backfat and abdominal fat from castrated and intact full-sib Yorkshire pigs was performed. The GO and KEGG enrichment analyses revealed that the host genes of the dorsal DE circRNAs were mainly involved in fatty acid transport, while in abdominal tissues, these genes were mainly involved in adipogenesis and inflammation. The interaction among sus_circPAPPA2, ssc-miR-2366 and GK was verified by dual fluorescence experiments and in porcine preadipocytes. The overexpression of sus_circPAPPA2 significantly inhibited the differentiation of preadipocytes. The expression of sus_circPAPPA2 was increased after adding 100 nM of testosterone, and preadipocyte differentiation was significantly inhibited. Testosterone can affect preadipocyte differentiation by upregulating the expression of sus_circPAPPA2, sponging miR-2366 and regulating the expression of genes, such as GK. These results indicate that testosterone can regulate the expression of adipocyte differentiation- and lipid metabolism-related genes by regulating the expression of circRNA, and ceRNA networks are different in the testosterone regulation of adipose deposition in different parts. This study provides basic data enhancing the understanding of the interaction between the hormone environment and mir-2366/GK to regulate trait performance in pigs.

P157. Role of the cecum on faecal microbial community structure in finisher pigs using a caecectomized pig model

The cecum is a major site of fermentation of dietary nutrients. In the pig, there is a knowledge gap pertaining to the role of the cecum in nutrient metabolism and health. The objective of this preliminary work was to develop a caecectomized pig model and to evaluate cecum microbial community structure in cecectomized finisher pigs fed a high fibre diet. Caecectomies were performed at the nursery to grower phase transition (∼ 63 d of age) prior to the experiment. A total of 10 barrows (initial BW = 83 kg; ∼112 d of age) were allotted to 1 of 2 treatments (5 replicates/treatment) in a 4 wk experiment. Treatments consisted of caecectomized (CEC-) or cecum intact (CTRL) pigs. All pigs were fed a moderately complex, high fibre (soy hulls included at 25%) finisher diet (3,006 ME kcal/kg) during the entirety of the trial. Data was analysed using PROC MIXED procedures of SAS with treatment as fixed effect and initial BW block as random effect. No differences were observed in BW on d 0 (82.92 vs 83.92 kg) or d 28 (110.78 vs 114.36 kg) between treatments (CEC- vs CTRL, respectively). During wk 4 CTRL pigs tended to have greater (P<0.09) ADFI compared to CEC- pigs (4.39 vs. 3.45 kg/d). Faecal samples (obtained on d 0, 14, and 28) were used to evaluate phylum-level bacterial community structure by sequencing the V4 region of the 16S rRNA gene. No significant differences were observed in microbial community structure between treatments over the time course of the experiment. Preliminary data suggests there may be utility in development of a caecectomized pig model to evaluate the role of the caecum in pig nutrient metabolism and health. Additional analyses are in process to investigate fibre digestion kinetics and bacterial community structure at the genus/species levels.

Influence of 2'-Fucosyllactose and Bifidobacterium longum Subspecies infantis Supplementation on Cognitive and Structural Brain Development in Young Pigs.

Development of the gut-brain axis during early-life is an important contributor of brain structural and functional development. Human milk oligosaccharides and gut microbiota have potential beneficial effects on various aspects of development; however, the effects of 2′-fucosyllactose (2′-FL) and Bifidobacterium longum subsp. infantis Bi-26 (Bi-26) administration during infancy separately and combined are still not clear. Therefore, we investigated the effects of early administration of dietary 2′-FL and Bi-26 on brain structural and functional development in the young pig. From postnatal day (PND) 2–34 or 35, fifty-two intact male pigs were randomly assigned to treatment groups in a 2 × 2 factorial arrangement and provided ad libitum access to a nutritionally adequate milk replacer without or with 1.0 g of 2′-FL/L of reconstituted liquid. Pigs within each diet group were further stratified to receive a daily oral dose of glycerol stock without or with Bi-26 (109 CFU). Pigs were subjected to the novel object recognition (NOR) task from PND 27–31 to assess recognition memory and subsequently underwent magnetic resonance imaging procedures at PND 32 or 33 to assess brain macrostructure and microstructure. Pigs that received Bi-26 had smaller absolute brain volumes for 9 of 27 brain regions of interest, and smaller relative volumes for 2 regions associated with kinesthesia (P < 0.05). Synbiotic administration of 2′-FL and Bi-26 elicited interactive effects (P < 0.05) on several microstructural brain components, where dual supplementation negated the effects of each test article alone. Behavioral outcomes indicated that pigs did not express novelty preference, regardless of treatment group, demonstrating no effects of 2′-FL and Bi-26 on recognition memory when supplemented alone or in combination. Interactive effects (P < 0.05) were observed for the number of all object visits, latency to the first object visit, and number of familiar object visits. Pigs that did not receive Bi-26 supplementation exhibited less time interacting with the familiar object in total (P = 0.002) and on average (P = 0.005). In conclusion, supplementation of 2′-FL and/or Bi-26 elicited some alterations in object exploratory behaviors and macro/micro-structures of the brain, but changes in recognition memory were not observed. Specifically in brain microstructure, synbiotic administration of 2′-FL and Bi-26 appeared to negate effects observed when each dietary article was supplemented separately.

Integrated Analysis of the ceRNA Network and M-7474 Function in Testosterone-Mediated Fat Deposition in Pigs.

Castration can significantly enhance fat deposition in pigs, and the molecular mechanism of fat deposition caused by castration and its influence on fat deposition in different parts of pigs remain unclear. RNA-seq was performed on adipose tissue from different parts of castrated and intact Yorkshire pigs. Different ceRNA networks were constructed for different fat parts. GO and KEGG pathway annotations suggested that testosterone elevates cell migration and affects differentiation and apoptosis in back fat, while it predisposes animals to glycolipid metabolism disorders and increases the expression of inflammatory cytokines in abdominal fat. The interaction between M-7474, novel_miR_243 and SGK1 was verified by dual fluorescence experiments. This ceRNA relationship has also been demonstrated in porcine preadipocytes. Overexpression of M-7474 significantly inhibited the differentiation of preadipocytes compared to the control group. When 100 nM testosterone was added during preadipocyte differentiation, the expression of M-7474 was increased, and preadipocyte differentiation was significantly inhibited. Testosterone can affect preadipocyte differentiation by upregulating the expression of M-7474, sponging novel-miR-243, and regulating the expression of genes such as SGK1. At the same time, HSD11B1 and SLC2A4 may also be regulated by the corresponding lncRNA and miRNA, which ultimately affects glucose uptake by adipocytes and leads to obesity.

The Novel-miR-659/SPP1 Interaction Regulates Fat Deposition in Castrated Male Pigs.

Simple SummaryCastration is a standard method for eliminating boar taint in industrial hog production, but it also causes enormous fat accumulation in the carcass. Secreted phosphoprotein 1 (SPP1) was selected to investigate its functions on the regulation of adipose deposition based on our previous data. In the present study, SPP1 overexpression and interference bidirectionally verified that SPP1 inhibited adipogenic differentiation of porcine bone marrow mesenchymal stem cells (pBMSCs). Testosterone-treated cell models were used to simulate the androgen status of intact pigs, and testosterone addition influenced SPP1 mRNA levels during the differentiation of pBMSCs. Moreover, we identified novel-miR-659 and targeted the 3′ untranslated region of SPP1 based on bioinformatics analysis and dual-luciferase assays, and found that the novel-miR-659 upregulation promoted adipogenesis while novel-miR-659 downregulation suppressed adipogenesis in pBMSCs detected by Oil Red O staining and adipogenic markers. Collectively, the interaction between novel-miR-659 and SPP1 can regulate adipose accumulation in castrated male pigs. Our data provide a theoretical basis for further study on the fat deposition mechanism caused by castration.Castration is usually used to remove boar taint in commercial pork production, but the adipose accumulation was increased excessively, which affected the meat quality of pigs. Based on our previous study, secreted phosphoprotein 1 (SPP1) was significantly differentially expressed between castrated and intact male pigs. However, the role of SPP1 in regulating adipose growth and fat storage caused by castration is unknown. In this study, SPP1 was identified to inhibit adipogenesis by the expression of adipogenic markers PPARγ and FABP4 as well as Oil red staining assay during differentiation of porcine bone marrow mesenchymal stem cells (pBMSCs). Subsequently, testosterone was used to treat pBMSCs to simulate the androgen status of intact pigs. Compared with the control groups without testosterone, the SPP1 expression in the testosterone group was markedly increased in the late stage of pBMSCs differentiation. Furthermore, novel-miR-659 was predicted by TargetScan and miRDB to target SPP1 and verified through a dual-luciferase reporter assay. Oil Red O staining assay indicated that novel-miR-659 overexpression significantly promoted adipogenesis, whereas novel-miR-659 inhibition suppressed adipogenesis. The expressions of adipogenic markers PPARγ and FABP4 showed the same tendency. Taken together, our study found that the targeted interaction between novel-miR-659 and SPP1 is involved in regulation of fat deposition in castrated male pigs.