Leydig Cells in Immunocastrated Polish Landrace Pig Testis: Differentiation Status and Steroid Enzyme Expression Status.

Porker immunocastration against gonadoliberin (GnRH) secretion has been utilized since 2009; however, consumers are still skeptical of it. This is due to not having full information available on the problem of a boar taint, as well as a lack of research on morphological and molecular changes that may occur in the animal reproductive system and other body systems. The present study aimed to explore the functional status of steroidogenic Leydig cells of the testicular interstitial tissue in immunocastrated Polish Landrace pigs. Analyses were performed using Western blot, immunohistochemistry for relaxin (RLN), insulin-like 3 protein (INSL3), pelleted growth factor receptor α (PDGFRα), cytochrome P450scc, 3ß- and 17ß-hydroxysteroid dehydrogenases (3ß-HSD, 17ß-HSD), cytochrome P450arom, and 5α-reductase (5α-RED). Immunoassay ELISA was used to measure the androstenone, testosterone, and estradiol levels in the testis and serum of immunocastrates. We revealed disturbances in the distribution and expression of (i) RLN, indicating an inflammatory reaction in the interstitial tissue; (ii) INSL3 and PDGFRα, indicating alterations in the differentiation and function of fetal, perinatal, or adult Leydig cell populations; (iii) P450scc, 3ß-HSD, 17ß-HSD, P450arom, and 5α-RED, indicating disturbances in the sex steroid hormone production and disturbed functional status of Leydig cells; as well as (iv) decreased levels of androstenone, testosterone, and estradiol in testicular tissue and serum, indicating the dedicated action of Improvac to reduce boar taint at both the hypothalamic-hypophysis-gonadal axis and local level (Leydig cells). In summary, our study provides a significant portion of knowledge on the function of Leydig cells after immunocastration, which is also important for the diagnosis and therapy of testis dysfunction due to GnRH action failure and/or Leydig cell differentiational-functional alterations.

The roles and expression of HOXA/Hoxa10 gene: A prospective marker of mammalian female fertility?

This review addresses the influence of homebox A10/a10 (HOXA/Hoxa10) gene on reproductive tract anatomy and functional fertility in mammalian species, and discusses major endocrine and environmental regulators of HOXA/Hoxa10 expression. Female reproductive efficiency or success is a function of several factors including the ovulation and fertilization rate, and uterine receptivity. A family of HOX/Hox genes establishes the segmental identity of the reproductive tract during embryogenesis and retains its physiological plasticity in sexually mature animals and humans. In particular, the HOXA/Hoxa10 gene is an intrinsic component of implantation, decidualization, and immunomodulation in the adult uterus. It was, therefore, suggested that knowledge of HOXA/Hoxa10 regulation might be essential in navigating molecular mechanisms with the aim of enhancing female reproductive potential. However, a recent study in pigs revealed a lack of associations between endometrial HOXA10 expression and reproductive tract morphology, and very poor correlations with sows' fertility metrics. Retinoic acid mainly regulates 3' HOX/Hox paralogs but may also modify the expression of downstream HOX/Hox genes, including HOXA/Hoxa10. Sex steroids directly regulate HOXA/Hoxa10 expression. The vitamin D receptor pathway modulates HOXA/Hoxa10 expression in the adult reproductive tract. Lastly, endocrine disruptors such as diethylstilbestrol, methoxychlor, bisphenol A, and isoflavones were shown to alter HOXA/Hoxa10 expression, thus affecting reproductive competence of the female.

Changes of Androstenone Concentrations in Saliva of Boars with Age.

With the increasing age of boars, the possibility of androstenone (5α-androst-16-en-3-one (AND), 5α-androst-16-en-3α-ol (α-AND), and 5α-androst-16-en-3ß-ol (ß-AND)) occurrence increases as well. The aim of this study was to evaluate concentrations of androstenone compounds in the saliva of boars concerning the age of animals. In total, 72 boars were evaluated (24 boars per replication). The effect of age (three different ages-152, 163, and 172 days) was observed, and (Landrace × Large White) × Pietrain genotypes were used. Chemical analysis of saliva samples was conducted by multidimensional gas chromatography/mass spectrometry (MDGC/MS). Salivary α-AND increased with age (p < 0.05), and positive correlations were found between age and submaxillary salivary gland weight (p < 0.05), age, and salivary AND concentration (p < 0.05), body weight and submaxillary salivary gland weight (p < 0.05), submaxillary salivary gland weight and salivary ß-AND concentration (p < 0.05), as well as submaxillary salivary gland weight and total salivary 5α-androstenone (AND total) concentration (p < 0.001). Nowadays, animal welfare is becoming a more and more discussed topic, and pig breeding is not an exception. Specifically, the castration ban is a current issue, so it is very important to know as much as possible about compounds responsible for boar taint. Androstenone's appearance in boars' saliva could be one of the most important precursors for future early detection of boar taint.

Correlates of reproductive tract anatomy and uterine histomorphometrics with fertility in swine.

Economic potential of the swine industry hinges upon the reproductive performance of sows, which may be enhanced by improving uterine capacity, a component trait of litter size and piglet productivity. Previous attempts at characterizing morphological traits indicative of high uterine volume have not been completely successful, resulting in the continued need for a reliable method of predicting reproductive value to improve production efficiency of the sow. Hence, the main objective of this study was to scrutinize macro- and micro-morphology of the sow's reproductive tract for quantitative correlations with fertility indices. Reproductive records from Polish Landrace × Polish Large White sows were used to examine the associations between fertility and ovarian/uterine morphology (n = 34) or uterine histomorphometry (n = 10). Several measures related to the ovary, including right and left ovarian weight (r = 0.50, p = 0.005 and r = 0.49, p = 0.006, respectively), were positively correlated with the litter size, while left ovarian number of corpora lutea (r = -0.38, p = 0.04) was negatively correlated with the mean litter size. Analysis of histomorphological characteristics of the uterine wall collected during the luteal phase of the estrous cycle revealed correlations between mean litter size and myometrial vascular content (r = 0.75, p = 0.03), the proportion of myometrial stroma (r = -0.68, p = 0.03), and the variability of endometrial thickness (r = -0.72, p = 0.02) in sows. Eight ovarian, vaginal and uterine characteristics were significantly correlated with mean lifetime numbers of live born and stillborn piglets/litter or the last litter size before slaughter. In conclusion, several anatomical and histomorphological metrics that relate to reproductive performance of swine may be used to inform production protocols and as a tool for selection of elite breeding sows, warranting future research into non-invasive or minimally invasive techniques for obtaining such measures.

Associations between Mammary Gland Echotexture and Milk Composition in Cows.

Thirty clinically healthy Holstein-Friesian cows underwent twice daily machine milking and ultrasonographic examinations of the udder just prior to and after milking. Digital ultrasonographic images of each udder quarter were subjected to computer-assisted echotextural analyses to obtain mean numerical pixel values (NPVs) and pixel heterogeneity (PSD) of the mammary gland parenchyma. The average milk yield and pH were higher (p < 0.05) in the morning, whereas crude fat, total solids, solids non-fat and citric acid content were higher (p < 0.05) during the evening milking period. Mean NPVs and PSDs of the mammary gland parenchyma were greater (p < 0.05) after than before milking. There were significant correlations among echotextural characteristics of the udder and protein percentage, lactose content and freezing point depression determined in the milk samples collected in the morning and crude protein, casein, lactose and solids non-fat in the evening. Our results can be interpreted to suggest that computerized analysis of the mammary gland ultrasonograms has the makings of a technique for estimating non-fat milk constituents in cows. However, future validating studies are necessary before this method can be employed in commercial settings and research. Moreover, significant inter-quarter differences in udder echogenicity may necessitate further echotextural studies of separate quarters.

Evaluation of SCD, ACACA and FASN Mutations: Effects on Pork Quality and Other Production Traits in Pigs Selected Based on RNA-Seq Results.

In recent years, pig producers have struggled with the problem of low intramuscular fat levels in pork, which impacts palatability and ultimately meat quality. Reduced levels of intramuscular fat are likely the result of breeding objectives aimed at increasing lean meat content. In this study, three mutations within candidate genes for fat content (SCD, ACACA, and FASN) were selected, based on RNA-seq results and the relationship between polymorphisms in genes related to lipid metabolism, fattening and slaughter characteristics, as well as pork quality, including IMF level, were evaluated to identify selection markers. Moreover, their impact on gene expression was also examined. The PCR-RFLP (polymerase cha- in reaction - restriction fragments length) method was used to establish genotypes and effect sizes of potential genetic markers were estimated using a GLM model. It was identified that a FASN missense variant was positively associated with the expression level of this gene, which suggested its linkage with a mutation having a regulatory function. The association study indicated that the FASN missense variant may play a role in the determination of feed conversion and meat colour. In turn, a mutation in the ACACA gene showed a relationship with IMF content in the Pulawska breed where the differences reached as much as 20%. We suggest considering all three mutations in further studies based on different pig populations due to the crucial role of SCD, ACACA, and FASN genes in lipid metabolism.

Association of Ghrelin Gene Polymorphisms with Fattening Traits and Feed Intake in Pig: A Preliminary Study.

Numerous studies have been conducted to explain the biological functions and mechanism of ghrelin (GHRL) action in animals. However, the exact role of ghrelin in the regulation of growth and development in pigs is still unclear. The ghrelin gene is considered to be a good candidate marker for the identification of economically important traits in pig production such as feed intake, growth or carcass quality. The objectives of the present study were to investigate the regulatory regions and coding sequence of the porcine GHRL gene and determine the effect of polymorphisms at the ghrelin gene locus on selected fattening traits. Data were obtained from 346 gilts (pure breeds: Landrace, 188; Duroc, 74; Pietrain, 84). The PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method was used to detect polymorphisms within GHRL. Three polymorphisms were found, one in the promoter region (c.-93A>G) and two in the 3'UTR sequence (g.4428T>C; g.4486C>T). A significant (p ≤ 0.01) additive effect on daily gain (negative) and age at slaughter (positive) was obtained at the locus c.-93A>G. However, the most promising mutation was at the locus g.4486C > T, which is associated with total feed intake. Overall, the described GHRL polymorphisms may be useful as molecular markers in pig selection but future studies are required.

Quantitative Echotextural Attributes of Pectoralis Major Muscles in Broiler Chickens: Physicochemical Correlates and Effects of Dietary Fat Source.

This study examined the relationships among physicochemical properties and ultrasonographic image attributes of pectoralis major muscles in broiler chickens. Forty male Ross 308 chicks were randomly assigned to four equinumerous fat-supplementation groups (Group SO: soybean oil; Group FO: flax oil; Group SO + FO: soybean oil + flax oil; and Group BF: beef fat). Ultrasonograms of birds' pectoral muscles were obtained just before slaughter at 6 weeks of age and were subjected to digital image analyses to determine the mean pixel intensity (MPI) and pixel heterogeneity values (standard deviation of numerical pixel values; MPH). A total of 2, 4, 2, and 6 significant correlations were recorded in Groups SO, FO, SO + FO, and BF, respectively; there were no correlations with the chemical composition of the muscles in Groups SO and SO + FO. The strongest correlations were found between muscle lightness (L*) and MPH in Group BF (physical characteristic; r = -0.82, p = 0.003), and between crude fat/protein content and MPI/MPH of pectoral the major muscles in Groups FO/BF (chemical characteristics; r = 0.72, p = 0.02). There exists a potential application of ultrasonographic imaging and computerized image analysis for predicting certain physicochemical properties of pectoralis major muscles in broiler chickens.

The Use of Vagina⁻Cervix Length Measurement in Evaluation of Future Reproductive Performance of Sows: A Preliminary Study under Commercial Conditions.

The length of the distal part of the internal reproductive tract seems to be related to the length and capacity of uterine horns, which is the most important anatomical property influencing litter size in sows. The aim of this study was to evaluate variation in vagina-cervix length (VCL) in gilts and differences in reproductive performance of sows according to VCL. The study was performed in a commercial farm using 221 gilts introduced into the breeding herd. Females were divided into three groups: (S) short (26.0 ± 2.0 cm, n = 36), (M) medium (31.3 ± 1.46 cm; n = 121), and (L) long VCL (36.0 ± 1.4 cm; n = 42) (p < 0.01). Mean live weight of gilts did not differ significantly among groups. Mean first litter size significantly varied between groups S (10.47 ± 3.01) and L (11.98 ± 2.32) (p = 0.0075) and M (10.67 ± 2.98) and L (p = 0.0054), while there was no significant difference between group S and M. Significant advantage (p = 0.023) was noted in the number of litters obtained from sows in groups L (4.69 ± 3.14), M (3.67 ± 2.71), and S (3.36 ± 2.40), and thus in total life production of sows (p = 0.0054), i.e., the number of piglets born alive. To conclude, the differences in vagina-cervix length in gilts during the first service was associated with significant variability in litter size during the first reproductive cycle, giving an advantage to females with longer VCL. Gilts with longer VCL were culled later and gave significantly more litters. Consequently, their lifetime piglet production was greater than gilts with shorter VCL.

Storage of boar semen at 16-18 °C in the long-term commercial extender prepared with deionized water or nanowater.

The aim of this study was to assess the usefulness of nanowater (NW; water declusterized using cold plasma treatment) as a diluent for a commercial boar semen extender during the 15-day storage (Days 1 to 15) at 16-18 °C. Ejaculates collected from 8 boars were subjected to the standard evaluation and then diluted in the extender prepared with deionized water (DW) or NW to a final concentration of 3×109 spermatozoa/ml. The proportion of defective spermatozoa increased (P<0.05) from Day 10 to Day 15 of storage (22.8±16.6% to 41.8±26.4% in DW group and 18.6±11.7% to 34.8±25.4% in NW group) and it was significantly greater in DW group compared with NW group on Days 5 and 10 due mainly to a greater (P<0.05) number of mid-piece defects in semen stored in the DW-containing extender. Sperm progressive motility decreased (P<0.05) in both groups between Days 2 and 6, Days 6 and 10, and Days 10 and 12, whereas the percentage of motile spermatozoa declined (P<0.05) to Day 14 only in NW group. Sperm motility was greater (P<0.05) in NW group compared with DW group from Day 5 to Day 13. A decline in sperm progressive motility below 40% in all semen samples occurred by Day 11 in DW group and by Day 12 in NW group. The mean survival time of sperm at 37 °C ex situ was greater in NW group than in DW group on Day 5 (314±87 min compared with 284±87 min) and Day 10 (223±34 min compared with 182±27 min; NW group compared with DW group, respectively). There were no differences (P>0.05) between the two groups in the concentrations of alkaline phosphatase and aspartate aminotransferase in semen extender. To summarize, the use of NW as an extender diluent exerts cytoprotective effects on boar spermatozoa and delays a decline in sperm progressive motility.