DigitalUM :: Browsing by Subject "Pig"

Browsing by Subject "Pig"

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Open Access
1H Nuclear Magnetic Resonance of pig seminal plasma reveals intra-ejaculate variation in metabolites
(MDPI, 2020-06-15) Mateo Otero, Yentel; Fernández-López, Pol; Gil-Caballero, Sergi; Fernandez-Fuertes, Beatriz; Bonet, Sergi; Barranco Cascales, Isabel; Yeste, Marc; Medicina y Cirugía Animal; Facultad de Veterinaria
In pigs, ejaculate is expelled in fractions, mainly the sperm-rich fraction (SRF) and the post-SRF (PSRF), which differ in both sperm content and origin. In addition, intra-ejaculate variability between fractions in terms of sperm reproductive characteristics has been previously reported, the highest sperm quality being observed in the first 10 mL of the SRF (SRF-P1). As seminal plasma (SP) composition has been purported to influence sperm physiology, the aim of this study was to profile pig SP metabolite composition and to find putative differences between the ejaculate portions (SRF-P1, the rest of SRF [SRF-P2], PSRF) and entire ejaculate (EE). To this end, ejaculates (n = 8, one per boar) were collected in fractions and SP was analyzed using 1H Nuclear Magnetic Resonance spectroscopy. We identified 19 metabolites present in all ejaculate portions and the EE, and reported correlations between the metabolites. Additionally, and for the first time in mammals, we found intra-ejaculate variability in the SP metabolites, observing different relative abundances in choline, glycerophosphocholine and glycine. Regarding their influence in sperm physiology, we hypothesize that these metabolites may explain the specific reproductive characteristics of each ejaculate portion. Finally, the reported SP metabolites could serve as a first steppingstone in the study of quality, functionality, and fertility biomarkers.
Open Access
Aldose reductase B1 in pig seminal plasma: identification, localization in reproductive tissues, and relationship with quality and sperm preservation
(Frontiers Media, 2021-06-08) Mateo Otero, Yentel; Viñolas Vergés, Estel; Llavanera, Marc; Ribas Maynou, Jordi; Roca, Jordi; Yeste, Marc; Barranco, Isabel; Medicina y Cirugía Animal
Aldose reductase B1 (AKR1B1), a NADPH-dependent enzyme that belongs to the aldo-keto reductase protein superfamily, has been reported to be involved in both male and female reproductive physiology. The objectives of this study were: (1) to evaluate the concentration of SP-AKR1B1 in pig ejaculate fractions; (2) to describe the immunohistochemical localization of AKR1B1 alongside the boar genital tract; (3) to evaluate the relationship between SP-AKR1B1 and sperm quality/functionality parameters. Ejaculates from seven boars (one ejaculate per boar) were collected in separate portions [the first 10 mL of the sperm rich fraction (SRF-P1), the rest of the SRF (SRF-P2), and the post-SRF (PSRF)], and the concentration of SP-AKR1B1 was assessed using an enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry and immunoblotting targeting was conducted in the reproductive tissues of these boars. Additionally, the entire ejaculates of 14 boars (one ejaculate per boar) were collected and split into three separate aliquots for: (i) SP-AKR1B1 quantification; (ii) assessment of sperm concentration and morphology; and (iii) evaluation of sperm quality and functionality parameters upon ejaculate collection (0 h) and after 72 h of liquid storage at 17°C. Concentration of AKR1B1 in the SP of SRF-P1 (458.2 ± 116.33 ng/mL) was lower (P < 0.05) than that of SRF-P2 (1105.0 ± 229.80 ng/mL) and PSRF (1342.4 ± 260.18 ng/mL). Monomeric and dimeric AKR1B1 forms were expressed alongside the reproductive tissues, except in the bulbourethral glands. No relationship between SP-AKR1B1 and sperm quality/functionality parameters was observed either at 0 h or after 72 h of storage at 17°C. In conclusion, AKR1B1 is expressed in the reproductive organs of boars (except bulbourethral glands) and a higher concentration is found in the PSRF suggesting that seminal vesicles would be the main secretory source. However, this enzyme does not appear to be related to sperm quality/functionality or to the sperm ability to withstand liquid storage at 17°C.
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Application of a score for evaluation of pain, distress and discomfort in pigs with lameness and prolapses: correlation with saliva biomarkers and severity of the disease
(2019-10) Contreras Aguilar, María Dolores; Escribano Tortosa, Damián; Martínez Miró, Silvia; López Arjona, Marina; Peres Rubio, Camila; Martínez Subiela, Silvia; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Medicina y Cirugía Animal
A score system was used to evaluate pain, distress and discomfort in healthy pigs and pigs with two different diseases: lameness and rectal prolapse. In addition, correlations between the results of this score and a panel of salivary biomarkers and severity of disease were studied. This panel included biomarkers of stress (cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) and lipase (Lip)), im- munity (adenosine deaminase isozymes 1 (ADA1) and 2 (ADA2)) and oxidative status (uric acid (UA), Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2)). Based on their score, diseased animals were subdivided in those without and with evident pain. Lame pigs and prolapsed pigs with pain showed higher salivary levels of cortisol, sAA, TEA, BChE, ADA1 and ADA2 compared with the healthy pigs. In addition, the prolapsed pigs with pain showed higher levels of FRAS, AOPP and H2O2 compared with the healthy animals. Salivary cortisol, TEA, BChE, ADA isozymes 1 and 2, FRAS and AOPP correlated with the pain score. This five-point pain score system can be easily applied to lame and prolapsed pigs, and salivary bio- markers could be used as an additional tool for pain assessment in those pigs.
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Changes in oxytocin concentrations in saliva of pigs after a transport andduring lairage at slaughterhouse
(Elsevier, 2020-08-31) López Arjona, Marina; Escribano, Damián; Mateo, Sandra V.; Contreras Aguilar, María Dolores; Pérez Rubio, Camila; Tecles Vicente, Fernando; Cerón Madrigal, José Joaquín; Martínez Subiela, Silvia; Medicina y Cirugía Animal
Oxytocin is associated with reproductive physiology but also with welfare and positive emotions. In this study,oxytocin was measured in saliva samples of 45 pigs that were collected before being transported to theslaughterhouse, at the time of arrival and 4 h after arrival to the slaughterhouse. Two previously validatedassays, one that measures free oxytocin and other that measures oxytocin linked to proteins, were used. Inaddition, cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) andlactate dehydrogenase (LDH), which are biomarkers associated with stress and pain in pigs, were measured. Theresults showed a decrease in free and protein-linked oxytocin concentrations at 4 h after transport comparedwith the time before transport, while cortisol, sAA, TEA, BChE and LDH showed an increase at 4 h after transportcompared with the time before transport. Based on these results it can be concluded that the transport andlairage at slaughterhouse in the conditions of this study produce a decrease in oxytocin in the saliva of pigs thatcould indicate a reduced emotional well-being
Open Access
Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction
(BMC, 2023-11-06) Viñolas Vergés, Estel; Ribas Maynou, Jordi; Barranco, Isabel; Peres Rubio, Camila; Bonet, Sergi; Roca, Jordi; Yeste, Marc; Medicina y Cirugía Animal
Background protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development. In some mammals, like pigs, ejaculates are emitted in three separate fractions: pre-sperm, sperm-rich (SRF) and post sperm-rich (PSRF). These fractions are known to vary in volume, sperm concentration and quality, as well as in the origin and composition of seminal plasma (SP), with differences being also observed within the SRF one. Yet, whether disparities in the DNA integrity and chromatin condensation and protamination of their sperm exist has not been interrogated. Results This study determined chromatin protamination (Chromomycin A3 test, CMA3), condensation (Dibromobi‑mane test, DBB), and DNA integrity (Comet assay) in the pig sperm contained in the frst 10 mL of the SRF (SRF-P1), the remaining portion of the sperm-rich fraction (SRF-P2), and the post sperm-rich fraction (PSRF). While chromatin protamination was found to be similar between the diferent ejaculate fractions (P>0.05), chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF (P=0.018 and P=0.004, respectively). Regarding DNA integrity, no diferences between fractions were observed (P>0.05). As the SRF-P1 has the highest sperm concentra‑tion and ejaculate fractions are known to difer in antioxidant composition, the oxidative stress index (OSi) in SP, calcu‑ lated as total oxidant activity divided by total antioxidant capacity, was tested and confrmed to be higher in the SRFP1 than in SRF-P2 and PSRF (0.42±0.06 vs. 0.23±0.09 and 0.08±0.00, respectively; P<0.01); this index, in addition, was observed to be correlated to the sperm concentration of each fraction (Rs=0.973; P<0.001). Conclusion While sperm DNA integrity was not found to difer between ejaculate fractions, SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF. This could be related to the OSi of each fraction.
Embargo
Circadian pattern of acute phase proteins in the saliva of growing pigs
(Elsevier, 2013-05) Gutiérrez Montes, Ana María; Escribano Tortosa, Damián; Fuentes Rubio, María; Cerón Madrigal, José Joaquín; Producción Animal
The circadian rhythm of the acute phase proteins (APPs) haptoglobin (Hp) and C-reactive protein (CRP) was assessed in saliva samples from 18- and 21-week old pigs. Saliva was collected at 07.00, 11.00, 15.00 and 19.00 h on two consecutive days and the Hp and CRP concentrations were quantified using two species-specific, time-resolved immunofluorometric assays. Salivary Hp levels were significantly higher (P < 0.001) in the morning compared to late afternoon (0.68 and 0.37 lg/mL, respectively) although the magnitude of the difference was much lower than is produced by inflammatory conditions. No significant differences were observed in CRP concentrations. Although the concentration of both APPs was higher in the 21- compared to the 18-week old pigs (P < 0.0001), no differences were observed in the circadian rhythm of these APPs when the two age groups were com- pared. Animal gender did not influence the circadian pattern of either APP, although the mean salivary CRP levels were higher in females (P < 0.05).
Open Access
Co-localization of the zinc transporter ZnT8 (slc30A8) with ghrelin and motilin in the gastrointestinal tract of pigs
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Schweiger, Markus; Steffl, Martin; Amselgruber, Werner M.
Zinc is an important co-factor for insulin storage in pancreatic β-cells of different species and the uptake of this ion into insulin containing secretory vesicles is managed by the zinc transporter, ZnT8, a member of the slc30A gene family. Recent studies indicate that this protein is a major autoimmune target in human type 1A diabetes and has also been implicated by genome-wide association studies in type 2 diabetes. Since individuals suffering from type 1 diabetes often develop gastrointestinal motility disorders, we investigated the expression of ZnT8 in the porcine gastrointestinal tract. For this purpose, we studied the cell-type specific expression of ZnT8 in the gut and its co-expression with endocrine hormones that are closely linked to intestinal motility regulation. Nested RT-PCR and immunostaining of sequential serial sections, as well as double-immunostaining using antibodies directed against ZnT8, ghrelin, motilin, neurotensin, serotonin and glucagon-like peptide 1, indicated that ZnT8 is colocalized with ghrelin and motilin. Our findings provide important information about the cell-type specific expression of ZnT8 in the porcine gastrointestinal system. The selective and exclusive expression of ZnT8 in two endocrine cell-types that are engaged in motility functions may be of particular interest for further investigations into type I diabetes-associated gastrointestinal dysfunctions.
Open Access
Data for compounds included in the article: "A sensitive immunoassay for the quantitation of Pig-MAP in pig saliva samples"
(2024-10-05) Piñeiro, M; Matas-Quintanilla, M; Miralles, A; Gutiérrez, AM; Medicina y Cirugía Animal
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Delays in processing and storage of pig seminal plasma alters levels of contained antioxidants
(2021-03) Barranco, Isabel; Tvarijonaviciute, Asta; Padilla, Lorena; Rodríguez-Martínez, Heriberto; Roca, Jordi; Xiomara, Lucas; Medicina y Cirugía Animal
Seminal plasma (SP) antioxidants are considered biomarkers of sperm function and fertility for AI-boars. The current protocol for their measurement implies the SP was harvested immediately after ejaculation and prompt stored at -80 °C until analysis. Such protocol may be impractical for AI-centers. This study evaluated how SP levels of antioxidants were influenced by delays in (1) SP-harvesting (0 [control], 2 or 24 h at 17 °C after ejaculate collection), in (2) SP-freezing (0 [control] or 24 h at 17 °C after SP-harvesting) or (3) the temperature of storage (-80 °C [control] or - 20 °C). The SP-antioxidants evaluated were: glutathione peroxidase [GPx], superoxide dismutase [SOD], paraoxonase-1 [PON-1], trolox equivalent antioxidant capacity [TEAC] and oxidative stress index [OSI]. A total of 120 aliquots from 10 entire ejaculates were handled in three trials. They were centrifuged (1500 g, 10 min) for harvesting SP and antioxidants were measured with an Automatic Chemistry Analyzer. A 24 h-delay in harvesting the SP led to an increase (p˂0.001) in TEAC and SOD SP-levels, and a decrease (p˂0.05) of OSI and PON-1. Similarly, a 24 h-delay to freeze the SP increased (p˂0.01) TEAC values and decreased (p˂0.01) PON-1 and GPx activity levels. Finally, storing the SP at -20 °C decreased (p˂0.001) SP-levels of TEAC, PON-1 and GPx, and increased (p˂0.01) OSI values. Strong positive relationships (p˂0.001) were found between antioxidant SP-levels in processed samples and their respective controls. In sum, handling and SP storage influence antioxidant measurements in AI-boars. Reliable levels of SP-antioxidants can only be warranted if a strict protocol for harvesting and SP storage is followed.
Open Access
Differences in weight, hierarchy, and incidence of lameness between two groups of adult pigs derived from assisted reproductive technologies
(MDPI, 2022-12-17) Ramírez, Lisette L.; Ortin Bustillo, Alba; Ramis, Guillermo; Romar, Raquel; Coy, Pilar; Romero Aguirregomezcorta, Jon; Ramírez, Lisette L.; Ortín Bustillo, Alba; Ramis Vidal, Manuel Guillermo; Romar Andrés, Raquel; Coy Fuster, Pilar; Anatomía y Anatomía Patológica Comparadas ; Facultad de Veterinaria
The in vitro production (IVP) and subsequent transfer of embryos (ET) to recipient mothers is not yet an established reproductive technology in the pig industry, as it is in cattle. However, that the trade of IVP-cryopreserved pig embryos is expected to start in the next decades. Society and governments are increasingly aware of the repercussions that IVP could have for animal health, welfare, behavior, or food safety, but proven scientific information for this type of animal does not exist, since no colonies of pigs have been created to this end. We created a small one and studied the differences between 16 IVP-derived pigs and 14 pigs derived from artificial insemination (AI), at 3.5 years of age, conceived from the same boar, and housed and fed under the same conditions since they were born. Incidence of lameness, position in the herd hierarchy, weight, adenosine deaminase activity, and hematological and biochemical analytes were compared between the two groups of animals. The results showed that the IVP animals weighed more, occupied higher positions in the herd hierarchy, and had a lower incidence of lameness. Although genetic differences from the maternal line could explain some of these results, it is also possible that the IVP animals developed better adaptative abilities, but more studies with a higher number of animals are necessary to reach consistent conclusions.
Open Access
Evaluation of S100A12 protein as a porcine health status biomarker when quantified in saliva samples.
Gutiérrez, A M; Matás-Quintanilla, M; Piñeiro, M; Sánchez, J; Fuentes, P; Medicina y Cirugía Animal
Datos de investigación relativos a un artículo de investigación: validación de un biomarcador, la proteína S100A12, de salud porcina, validación analítica y clínica de un ensayo para cuantificar las concentraciones del biomarcador en muestras de saliva porcina.
Open Access
Exploring seminal plasma GSTM3 as a quality and in vivo fertility biomarker in pigs-relationship with sperm morphology
(MDPI, 2020-08-12) Llavanera, Marc; Delgado-Bermúdez, Ariadna; Mateo-Otero, Yentel; Padilla, Lorena; Romeu, Xavier; Roca, Jordi; Barranco, Isabel; Yeste, Marc; Medicina y Cirugía Animal
Glutathione S-transferases Mu 3 (GSTM3) is an essential antioxidant enzyme whose presence in sperm has recently been related to sperm cryotolerance, quality and fertility. However, its role in seminal plasma (SP) as a predictor of the same sperm parameters has never been investigated. Herein, cell biology and proteomic approaches were performed to explore the presence, origin and role of SP-GSTM3 as a sperm quality and in vivo fertility biomarker. GSTM3 in SP was quantified using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit specific for Sus scrofa, whereas the presence of GSTM3 in testis, epididymis and accessory sex glands was assessed through immunoblotting analysis. Sperm quality and functionality parameters were evaluated in semen samples at 0 and 72 h of liquid-storage, whereas fertility parameters were recorded over a 12-months as farrowing rate and litter size. The presence and concentration of GSTM3 in SP was established for the first time in mammalian species, predominantly synthesized in the epididymis. The present study also evidenced a relationship between SP-GSTM3 and sperm morphology and suggested it is involved in epididymal maturation rather than in ejaculated sperm physiology. Finally, the data reported herein ruled out the role of this antioxidant enzyme as a quality and in vivo fertility biomarker of pig sperm.
Open Access
Expression of transforming growth factor-B3 -GF-B3- in the porcine ovary during the oestrus cycle
(Murcia : F. Hernández, 2008) Steffl, M.; Schweiger, M.; Amselgruber, W.
Transforming growth factor-ß (TGF-ß) proteins are growth factors that have been shown to be involved in regulation of ovarian follicular development. Ovarian expression, activity and functional significance of TGF-ß1 and TGF-ß2 isoforms were extensively studied in most species. However, little is known about the biological role of TGF-ß3 previously shown to be expressed independently of the other two isoforms. Therefore, expression of TGF-ß3 mRNA and protein was evaluated by RT-PCR and immunohistochemistry, respectively, in porcine ovaries collected during different phases of the oestrus cycle. Results of RT-PCR analysis showed that TGF-ß3 mRNA is expressed throughout the oestrus cycle. The level of TGF-ß3 mRNA expression was found to be higher at metoestrus and dioestrus. Weak TGF-ß3 immunoreactivity was present in follicular epithelial cells and oocytes of preantral follicles in all stages examined. TGF-ß3 protein expression was exclusively present in theca interna cell layer of antral follicles, and was particularly prominent in large antral follicles. Immediately after ovulation, almost all theca cells outside of the granulosa cell layer were intensively stained with anti-TGF-ß3. Immunostaining of TGF-ß3 in theca lutein cells rapidly decreased during corpus luteum development. It is suggested that TGF-ß3 may play an important role in modulating theca cell function of pre- and postovulatory follicles of the pig.
Open Access
Extracellular vesicles isolated from porcine seminal plasma exhibit different tetraspanin expression profiles
(2019-08-09) Barranco, Isabel; Padilla, Lorena; Parrilla, Inmaculada; Alvarez-Barrientos, Alberto; Pérez-Patiño, Cristina; Peña, Fernando; Martínez, Emilio A; Rodriguez-Martínez, Heriberto; Roca, Jordi; Roca, Jordi; Medicina y Cirugía Animal
Seminal extracellular vesicles (EVs) include exosomes (ø 40-120 nm) and microvesicles (MVs, ø 120-1000 nm), which would be involved in multiple functional reproductive roles. The study aimed to establish which EV subtypes are present in pig semen, using a high-resolution flow cytometer to explore differences in their tetraspanin expression profile. The EVs were isolated from 12 pig ejaculates using serial ultracentrifugation and characterized by dynamic light scattering and electron microscopy for size and morphology as well as for tetraspanin expression using flow cytometry with Carboxyfluorescein succinimidyl ester (CFSE) and antibodies against CD9, CD63 and CD81. Pig semen contained a heterogeneous EV-population regarding size and morphology. Flow cytometric analysis demonstrated that the proportion of EVs expressing CD63 and CD9 was higher in MVs (P < 0.001 and P < 0.05, respectively) than in exosomes, while the opposite was true for CD81; higher (P < 0.001) in exosomes than in MVs. In conclusion, (1) the new generation of flow cytometers are able to accurately identify EVs and to gate them in two size-different populations named exosomes and MVs. (2) Tetraspanins CD9, CD63 and CD81 are present in both seminal EVs, albeit with exosomes and MVs differing in expression profiles, suggesting dissimilar cargo and binding affinity.
Open Access
Flutamide alters β-catenin expression and distribution, and its interactions with E-cadherin in the porcine corpus luteum of mid- and late pregnancy
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2015) Grzesiak, Malgorzata; Mitan, Agata; Janik, Marcelina E.; Knapczyk-Stwora, Katarzyna; Slomczynska, Maria
This study examined whether flutamideinduced androgen deficiency during mid- and late pregnancy in pigs affected luteal expression of adherens junction protein, β-catenin, and its interactions with Ecadherin. Flutamide (50 mg/kg body weight) was administered into pregnant gilts between days 43-49 (GD50F), 83-89 (GD90F) or 101-107 (GD108F) of gestation. Corpora lutea (CLs) were obtained on day 50, 90 or 108 of pregnancy (n=8-11 per each group). Total β-catenin and E-cadherin expression was examined at mRNA (real-time PCR) and protein (Western blot) level. Moreover, subcellular β-catenin fractions were extracted and immunoblotted. Immunohistochemistry was used for β-catenin localization. To determine whether flutamide disturbs β-catenin/E-cadherin mutual interactions, coimmunoprecipitation using anti-β-catenin antibody was performed. Furthermore, phosphorylation of Ecadherin was assessed. Flutamide exposure led to decreased β-catenin mRNA expression in all examined groups (p<0.001 or p<0.01), but protein level was lower only in the GD90F and GD108F groups (p<0.05). Ecadherin mRNA (p<0.05 or p<0.01) and protein (p<0.05) levels were up-regulated in all flutamidetreated groups when compared to controls. β-catenin was predominantly found in membranes of luteal cells with no significant changes after antiandrogen treatment. βcatenin/E-cadherin complexes were more abundant in the GD90F (p<0.05) and GD108F (p<0.01) groups than in controls due to enhanced E-cadherin phosphorylation at serine 838/840 in those animals (p<0.05). Overall, although androgen deficiency affected β-catenin expression in the CL of pregnancy in pigs, a compensatory mechanism by enhanced interactions with E-cadherin is possible. Thus, androgen signaling via androgen receptors appears to be crucial in the regulation of luteal cells cross-talk.
Open Access
High total antioxidant capacity of the porcine seminal plasma (SP-TAC) relates to sperm survival and fertility
(2015-12-21) Barranco, Isabel; Tvarijonaviciute, Asta; Perez-Patiño, Cristina; Parrilla, Inmaculada; Cerón, Jose J; Martínez, Emilio A; Rodriguez-Martinez, Heriberto; Roca, Jordi; Medicina y Cirugía Animal
The study attempted to clarify the role of total antioxidant capacity of seminal plasma (SP-TAC) on boar sperm survival and fertility after artificial insemination (AI). SP-TAC differed (P < 0.001) among boars (n° = 15) and, to a lesser degree, among ejaculates within male (4 ejaculates/boar). SP-TAC also differed (P < 0.001) among ejaculate fractions (43 ejaculates and 3 fractions per ejaculate), of which the sperm-peak portion of the sperm rich ejaculate fraction (SRF) had the highest SP-TAC. SP-TAC was not correlated with sperm quality (motility and viability) or functionality (intracellular ROS generation and lipid peroxidation) of liquid AI-semen samples stored at 17 °C for 72 h (90 AI-samples), but the decline in sperm quality was larger (P < 0.05) in ejaculates with low, compared with high SP-TAC (hierarchically grouped). The SP-TAC differences among ejaculate portions agree with sperm cryosurvival rates (14 ejaculates from 7 boars), showing sperm from sperm-peak portion better (P < 0.01) post-thaw quality and functionality than those from the entire ejaculate (mainly post-SRF). Boars (n° = 18) with high SP-TAC (hierarchically grouped) had higher (P < 0.05) fertility outcomes (5,546 AI-sows) than those with low SP-TAC. Measurement of SP-TAC ought to be a discriminative tool to prognosis fertility in breeding boars.
Open Access
Immunophenotype profile by flow cytometry reveals different subtypes of extracellular vesicles in porcine seminal plasma
(2024-01-23) Barranco, Isabel; Alvarez-Barrientos, Alberto; Parra, Ana; Martinez-Diaz, Pablo; Lucas, Xiomara; Roca, Jordi; Medicina y Cirugía Animal
Background: Porcine seminal plasma (SP) is endowed with a heterogeneous population of extracellular vesicles (sEVs). This study evaluated the immunophenotypic profile by high-sensitivity flow cytometry of eight sEV subpopulations isolated according to their size (small [S-sEVs] and large [L-sEVs]) from four different SP sources, namely three ejaculate fractions (the first 10 mL of the sperm rich fraction [SRF-P1], the remaining SRF [SRF-P2], and the post-SRF [PSRF]) and entire ejaculate (EE). Methods: Seminal EVs were isolated using a size exclusion chromatography-based protocol from six SP pools (five ejaculates/pool) of each SP source and characterized using complementary approaches including total protein (BCA™assay), particle size distribution (dynamic light scattering), morphology (transmission electron microscopy), and purity (albumin by Western blot). Expression of CD9, CD63, CD81, CD44 and HSP90β was analyzed in all sEV subpopulations by high-sensitivity flow cytometry according to MIFlowCyt-EV guidelines, including an accurate calibration, controls, and discrimination by CFSE-labelling. Results: Each sEV subpopulation exhibited a specific immunophenotypic profile. The percentage of sEVs positive for CD9, CD63, CD81 and HSP90β differed between S- and L-sEVs (P < 0.0001). Specifically, the percentage of sEVs positive for CD9 and CD63 was higher and that for CD81 was lower in S- than L-sEVs in the four SP sources. However, the percentage of HSP90β-positive sEVs was lower in S-sEVs than L-sEVs in the SRF-P1 and EE samples. The percentage of sEVs positive for CD9, CD63, and CD44 also differed among the four SP sources (P < 0.0001), being highest in PSRF samples. Notably, virtually all sEV subpopulations expressed CD44 (range: 88.04-98.50%). Conclusions: This study demonstrated the utility of high-sensitivity flow cytometry for sEV immunophenotyping, allowing the identification of distinct sEV subpopulations that may have different cellular origin, cargo, functions, and target cells.
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Improvement of boar sperm cryosurvival by using single-layer colloid centrifugation prior freezing
(Elsevier, 2012-09-15) Martínez Alborcia, M. J.; Morrell, Jane M.; Parrilla Riera, Inmaculada; Barranco Cascales, Isabel; Vázquez Autón, José María; Martínez García, Emilio; Roca Aleu, Jorge; Medicina y Cirugía Animal
The aim of this experimental study was to evaluate the effectiveness of sperm selection using single-layer centrifugation (SLC) prior to freezing on the sperm cryosurvival of boar ejaculates. Twenty-four sperm rich ejaculate fractions (SREF), collected from 24 boars (one per boar), were divided into two groups according to their initial semen traits: standard (n = 15) and substandard (n = 9). Semen samples from each SREF were split in two aliquots, one remained untreated (control samples) and the other was single-layer centrifuged (500 g for 20 min) using 15 mL of Androcoll-P Large (SLC samples). The yield of total, motile (assessed by CASA) and viable (cytometrically evaluated after staining with H-42, propidium iodide (PI) and FITC-PNA) sperm after SLC was higher (P < 0.05) in standard than substandard semen samples. The semen samples were cryopreserved using a standard 0.5-mL straw freezing protocol. Post-thaw sperm motility and viability (assessed at 30 and 150 min post-thawing) were higher (P < 0.05) in SLC than in control samples, regardless of the initial semen traits of the ejaculates. Additionally, thawed spermatozoa from SLC samples were more resistant (P < 0.05) to lipid peroxidation (BIOXYTECH MDA-586 Assay Kit) than those from control samples, regardless of the initial semen traits of the ejaculates. The SLC-treatment also influenced the functionality of thawed spermatozoa undergoing an in vitro capacitation process. The percentage of viable sperm showing high membrane fluidity (assessed with merocyanine 540) was lower (P < 0.05) in the SLC than in the control samples, regardless of the initial semen traits of the ejaculates. Thawed viable spermatozoa of SLC samples generated less (P < 0.05) reactive oxygen species (assessed with CM-H(2)DCFDA) than those of control samples in the substandard ejaculates. These findings indicate that the sperm selection before freezing using SLC improves the freezability of boar sperm.
Open Access
Influence of the circadian cycle, sex and production stage on the reference values of parameters related to stress and pathology in porcine saliva
Gutiérrez, A M; Saco, Y; Peña, Rl; Matas-Quintanilla, M; Ibáñez-López, F J; Piñeiro, M; Sotillo, J; Bassols, A; Medicina y Cirugía Animal
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Insulin in the saliva of pigs: Validation of an automated assay and changes at different physiological conditions
(2021-10-20) Ortín-Bustillo, Alba; Tvarijonaviciute, Asta; Cerón, José Joaquín; Contreras-Aguilar, María Dolores; Martínez-Miró, Silvia; Martínez-Subiela, Silvia; Tecles, Fernando; Muñoz-Prieto, Alberto; Medicina y Cirugía Animal
This study aimed to evaluate whether insulin could be measured in the saliva of pigs and if its concentration changes in some physiological conditions. For this purpose, a validation of an automated heterologous immunoassay for measuring insulin in the saliva of pigs was performed. In addition, the possible changes of salivary insulin concentration in sows after food intake and during gestation and lactation were studied. The evaluated immunoassay was able to detect insulin in the saliva of pigs in a precise and accurate way when species-specific calibrators were used. There was no correlation in insulin concentrations between serum and saliva. Insulin concentrations showed a significant increase in the saliva of sows after feeding. Sows at farrowing and lactation presented higher salivary insulin levels as compared with those in gestation. In conclusion, the results showed that insulin could be measured in the saliva of pigs, and changes in its concentration can be detected due to food intake and different physiological conditions.

Browsing by Subject "Pig"

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