Browsing by Subject "Assisted reproductive technologies"
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- PublicationOpen AccessGrowth parameters and growth-related hormone profile in a herd of cattle up to 4 years of age derived from assisted reproductive technologies(MDPI, 2025-02-21) Heras, Sonia; Sena Lopes, Jordana; Quintero-Moreno, Armando; Romero-Aguirregomezcorta, Jon; Canovas, Sebastian; Romar, Raquel; Coy, Pilar; Fisiología
- PublicationEmbargoGrowth performance in pigs derived from in vitro produced embryos is enhanced compared to their artificial insemination-derived counterparts from birth to adulthood(Elsevier, 2025) Heras, Sonia; Soriano-Ubeda, Cristina; Quintero-Moreno, Armando; Romero-Aguirregomezcorta, Jon; Paris-Oller, Evelyne; Gadea, Joaquin; Romar, Raquel; Canovas, Sebastian; Coy, Pilar; FisiologíaResearch suggests that assisted reproductive technologies may lead to adverse effects on the offspring. To date, long-term effects from fetal development through adulthood have been scarcely researched. In the present observational study, we aimed to describe growth (body weight, BW; crown-rump length, CRL; and average daily gain, ADG), and growth-related hormone levels (growth hormone, GH; insulin-like growth factor 1, IGF-1; thyroxine, T4; cortisol) of pigs derived from artificial insemination (AI) and from embryos produced in vitro under two different conditions, C-IVP, with Bovine Serum Albumin (BSA), and RF-IVP, with reproductive fluids and BSA, as protein source in culture, from birth to 5 years of age. In vitro-produced animals showed greater BW, CRL and ADG than AI. Additionally, C-IVP animals had greater BW, CRL and ADG than RF-IVP up to 6 months of age. Regarding hormones, GH concentration was greater in RF-IVP compared to AI and C-IVP. In males, IGF-1 levels of RF-IVP were greater than those of AI. Instead, T4 levels of AI males were greater than C-IVP and RFIVP. Males showed greater IGF-1 and GH than females, the latter only from 1 year old. Instead, females had greater T4 concentration than males. No differences in cortisol were observed between groups or sexes. In conclusion, this study sets reference values of growth and hormone concentrations for adult pigs that are missing in literature. Although the differences observed between the experimental groups were within physiological ranges, there was a greater growth performance in the IVP groups.
- PublicationOpen AccessLong term study of the blood plasma biochemical profile of cattle born by assisted reproductive technologies(Brazilian College of Animal Reproduction, 2024) Serrano-Albal, Maria; Romero-Aguirregomezcorta, Jon; Canovas, Sebastian; Heras, Sonia; Quintero-Moreno, Armando Arturo; Lopes, Jordana Sena; Soriano-Ubeda, Cristina; Gadea, Joaquin; Coy, Pilar; Romar, Raquel; FisiologíaAssisted reproductive technology (ART) is key in cattle breeding as it enables selective mating and improves both reproductive efficiency and genetic improvement. However, there is still limited information on the long-term effects of these methods on the health and development of the offspring. This study aims to investigate the biochemical profile of cattle born through ART, from a previous study (Lopes et al., 2020), over an extended period to understand how these techniques may affect their health and metabolism. Male and female (not pregnant or lactating) cattle born after artificial insemination (AI, N= 7) were compared to those derived from in vitro produced embryos (IVP) using a standard protocol (C-IVP, N= 7), or using oviductal and uterine fluids in the process (RF-IVP, N= 4). Males and females were kept in two diferent open pens, but fed and managed under identical conditions their whole lives. Animals were studied every six months, from 1.5 years old until ≥4 years old, since their first year of life was evaluated by Lopes et al. (2022). Tail vein blood plasma was obtained by centrifugation (1000 G, 10 min) in lithium heparin tubes and stored at 80°C. The biochemical profile included total protein (TP), albumin (ALB), globulin (GLOB), creatinine (CREA), urea (URE), glucose (GLUC), cholesterol (CHOL), triglycerides (TRIG), amylase (AMIL), lipase (LIP), creatinine kinase (CK), alkaline phosphatase (ALP), gamma-glutamyl ranspeptidase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TB). After serial dilutions, inter- and intraassay precision of the methods were lineal and below 15%. Data normal ly distributed was analysed using ANOVA and Tukey’s test; data not normally distributed was analysed using the Kruskal Wallis and Games Howell’s test (statistical significance p<0.05). The data is shown below as means by group over the years. The AI group had lower CREA and CHOL levels (1.43±0.05 mg/dL and 109.97±5.27 mg/dL, respectively) than C-IVP (1.75±0.07 mg/dL and 126.57±7.53 mg/dL, respectively) and RF-IVP (1.86±0.08 mg/dL and 140.01±7.19 mg/dL, respectively). The GGT levels were lower in RFIVP animals (13.00±1.30 UI/L) than both the C-IVP (17.55±1.10 UI/L) and AI groups (16.05±1.23 UI/L). Moreover, the levels of ALT were significantly lower in C-IVP (28.60±1.72 UI/L) than in the AI (34.35±1.81 UI/ L) and RF-IVP groups (33.40±2.24 UI/L). Depending on the age of the animal at the time of sample collection, significant differences were observed for most parameters. The parameters TP, GLOB, CREA, URE, AMIL, AST an increased in their concentration was observed as the animals aged, whilst ALB, GLUC, ALP decreased with age. However, all values can be considered within physiological ranges despite the significant v ariations. To conclude, the study showed differences in some biochemical parameters amongst the ART groups and with aging. However, despite the diferences all values were within physiological ranges. These findings are key to study the potential implications of this diferences on the animal’s health.
- PublicationOpen AccessLong-term study of physical, haematological and biochemical parameters in cattle with different embryo origins(MDPI, 2025-06-14) Serrano-Albal, Maria; Romero-Aguirregomezcorta, Jon; Canovas, Sebastian; Heras, Sonia; Gadea, Joaquin; Coy, Pilar; Romar, Raquel; FisiologíaAssisted reproductive technologies are vital in cattle breeding to improve genetic selection and productivity. While early-life differences between artificially inseminated (AI) and in vitro-produced (IVP) cattle have been studied, long-term physiological, haematological, and biochemical effects remain unclear. This observational study assessed AI and IVP cattle from 1.5 to 5 years of age to determine if early differences persist. IVP cattle were produced after the transfer of the embryo produced by supplementing (RF-IVP group) or not supplementing (C-IVP) the embryo culture with oviductal and uterine fluids. Physical evaluations showed body mass index increased until 3.5 years, while temperature and respiratory rate declined with age, with no significant differences between AI and IVP groups. Haematological analysis revealed age-related changes, including decreased red and white blood cell counts and increased mean corpuscular volume and haemoglobin. AI cattle had higher white blood cell counts than IVP groups. Sex significantly influenced many haematological variables. Biochemical analysis showed age-related increases in total protein, creatinine, and urea, and decreases in glucose and alkaline phosphatase. AI cattle had lower cholesterol and creatinine than IVP groups. Despite group differences, all values remained within normal ranges. Sex affected albumin, cholesterol, triglycerides, and creatine kinase. This study provides the first long-term haematological and biochemical reference values for cattle from different reproductive methods, showing that age is the main influencing factor and supporting IVP cattle as a viable alternative to AI in breeding programs.
- PublicationOpen AccessMetabolic profile and glycemic response in fully-grown sows born using assisted reproductive technologies(Elsevier, 2024-10-02) Canovas, Sebastian; Heras, Sonia; Romero-Aguirregomezcorta, Jon; Quintero-Moreno, Armando; Gadea, Joaquin; Coy, Pilar; Romar, Raquel; FisiologíaThe aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (P < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.
- PublicationOpen AccessMetabolic profile and glycemic response in fully-grown sows born using assisted reproductive technologies(Elsevier, 2024-10-02) Cánovas, Sebastián; Heras, S.; Romero-Aguirregomezcorta, J.; Quintero-Moreno, A.A.; Gadea, Joaquín; Coy, P.; Romar, Raquel; FisiologíaThe aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (P < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.
- PublicationOpen AccessReproductive fluids, added to the culture media, contribute to minimizing phenotypical differences between in vitro-derived and artificial insemination-derived piglets(Cambridge University Press, 2022) París-Oller, Evelyne; Soriano-Úbeda, Cristina; Belda-Pérez, Ramsés; Sarriás-Gil, Lucía; Canha-Gouveia, Analuce; Gadea, Joaquín; Vieira, Luis Alberto; García-Vázquez, Francisco Alberto; Romar, Raquel; Cánivas, Sebastián; Coy, Pilar; FisiologíaThe addition of reproductive fluids (RF) to the culture media has shown benefits in different embryonic traits but its long-term effects on the offspring phenotype are still unknown. We aimed to describe such effects in pigs. Blood samples and growth parameters were collected from piglets derived from in vitro-produced embryos (IVP) with or without RF added in the culture media versus those artificially inseminated (AI), from day 0 to month 6 of life. An oral glucose tolerance test was performed on day 45 of life. We show here the first comparative data of the growth of animals produced through different assisted reproductive techniques, demonstrating differences between groups. Overall, there was a tendency to have a larger size at birth and faster growth in animals derived from in vitro fertilization and embryo culture versus AI, although this trend was diminished by the addition of RFs to the culture media. Similarly, small differences in hematological indices and glucose tolerance between animals derived from AI and those derived from IVP, with a sex-dependent effect, tended to fade in the presence of RF. The addition of RF to the culture media could contribute to minimizing the phenotypical differences between the in vitro-derived and AI offspring, particularly in males.
- PublicationOpen AccessReproductive fluids, used for the in vitro production of pig embryos, result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM(BMC, 2021) París-Oller, E.; Navarro-Serna, S.; Soriano-Úbeda, C.; Lopes, J.S.; Matás, C.; Ruiz, S.; Latorre, L.; López-Albors, O.; Romar, R.; Cánovas, S.; Coy, P.; FisiologíaBackground: In vitro embryo production (IVP) and embryo transfer (ET) are two very common assisted reproductive technologies (ART) in human and cattle. However, in pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow. In addition, the potential impact of these two ART on the health of the offspring is unknown. We investigated here if the use of a modified IVP system, with natural reproductive fluids (RF) as supplements to the culture media, combined with a minimally invasive surgery to perform ET, affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits. Results: The blastocyst rates obtained by both in vitro systems, conventional (C-IVP) and modified (RF-IVP), were similar. Pregnancy and farrowing rates were also similar. However, when compared to in vivo control (artificial insemination, AI), litter sizes of both IVP groups were lower, while placental efficiency was higher in AI than in RF-IVP. Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI, but not for RF-IVP group. Conclusions: The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring.
- PublicationOpen AccessReproductive fluids, used for the in vitro production of pig embryos, result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM.(BMC, 2021-02-15) París-Oller, E.; Navarro-Serna, S.; Soriano-Ubeda, C.; Lopes, J.S.; Matás, C.; Ruiz, S.; Latorre, R.; López-Albors, O.; Cánovas, S.; Coy, P.; Romar, R.; Fisiología; Anatomía y Anatomía Patológica ComparadasBackground: In vitro embryo production (IVP) and embryo transfer (ET) are two very common assisted reproductive technologies (ART) in human and cattle. However, in pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow. In addition, the potential impact of these two ART on the health of the offspring is unknown. We investigated here if the use of a modified IVP system, with natural reproductive fluids (RF) as supplements to the culture media, combined with a minimally invasive surgery to perform ET, affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits. Results: The blastocyst rates obtained by both in vitro systems, conventional (C-IVP) and modified (RF-IVP), were similar. Pregnancy and farrowing rates were also similar. However, when compared to in vivo control (artificial insemination, AI), litter sizes of both IVP groups were lower, while placental efficiency was higher in AI than in RF-IVP. Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI, but not for RF-IVP group. Conclusions: The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring. Keywords: Assisted reproductive technologies, Embryo transfer, In vitro embryo production, Placenta, Reproductive fluids
- PublicationOpen AccessSpontaneous and ART-induced large offspring syndrome: similarities and differences in DNA methylome(Taylor and Francis Group, 2022-05-03) Yahan Li; Sena Lopes, Jordana; Coy-Fuster, Pilar; Melissa Rivera, Rocío; Didáctica y Organización EscolarLarge/abnormal offspring syndrome (LOS/AOS) is a congenital overgrowth syndrome reported in ruminants produced by assisted reproduction (ART-LOS) which exhibit global disruption of the epigenome and transcriptome. LOS/AOS shares phenotypes and epigenotypes with the human congenital overgrowth condition Beckwith-Wiedemann syndrome. We have reported that LOS occurs spontaneously (SLOS); however, to date, no study has been conducted to determine if SLOS has the same methylome epimutations as ART-LOS. In this study, we performed whole- genome bisulphite sequencing to examine global DNA methylation in bovine SLOS and ART-LOS tissues. We observed unique patterns of global distribution of differentially methylated regions (DMRs) over different genomic contexts, such as promoters, CpG Islands, shores and shelves, as well as at repetitive sequences. In addition, we included data from two previous LOS studies to identify shared vulnerable genomic loci in LOS. Overall, we identified 320 genomic loci in LOS that have alterations in DNA methylation when compared to controls. Specifically, there are 25 highly vulnerable loci that could potentially serve as molecular markers for the diagnosis of LOS, including at the promoters of DMRT2 and TBX18, at the imprinted gene bodies of IGF2R, PRDM8, and BLCAP/NNAT, and at multiple CpG Islands. We also observed tissue-specific DNA methylation patterns between muscle and blood, and conservation of ART-induced DNA methyla-tion changes between muscle and blood. We conclude that as ART-LOS, SLOS is an epigenetic condition. In addition, SLOS and ART-LOS share similarities in methylome epimutations.