Gastroprotective role of a flavonoid-rich subfraction from Fridericia chica (Bonpl.) L. G. Lohmann: a medicinal plant used in the Amazon region.

Fridericia chica is an Amazonian plant used to treat stomach disorders. However, the pharmacological activity of flavonoids in the extract has yet to be investigated. Therefore, we considered that a flavonoid-rich F. chica subfraction (FRS) has gastroprotective functions. For this, before the induction of gastric ulcers with ethanol or piroxicam, the rats received vehicle (water), omeprazole (30 mg/kg), or FRS (30 mg/kg), and the ulcer area was measured macro and microscopically, and the antisecretory action was investigated in pylorus-ligated rats. In addition, the roles of nitric oxide (NO) and nonprotein sulfhydryl compounds (NP-SH) in the gastroprotective effects of FRS were studied. FRS reduced ethanol- and piroxicam-induced ulcerations by 81% and 77%, respectively, as confirmed histologically. Antioxidant effects were observed for FRS through the maintenance of GSH and LPO levels, and the SOD and CAT activity similar to those found in the nonulcerated group. Moreover, FRS avoided the increase in MPO activity and TNF, IL-6, IL-4 and IL-10 levels. Moreover, mucin staining increased in ulcerated rats receiving FRS, and the pharmacological mechanism gastroprotective seems to involve the NO and NP-SH in addition to antisecretory actions. The chemical study by mass spectrometry confirmed the presence of flavonoids in FRS, and molecular docking studies have shown that these compounds interact with cyclooxygenase-1 and NO synthase. Furthermore, there was no indication that FRS had cytotoxic effects. Our results support the popular use of F. chica, and we conclude that the gastroprotection effect promoted by FRS can be attributed to the combined effect of the flavonoids.

Genome transfer technique for bovine embryo production using the metaphase plate and polar body.

Despite many studies in humans and mice using genome transfer (GT), there are few reports using this technique in oocytes of wild or domestic animals. Therefore, we aimed to establish a GT technique in bovine oocytes using the metaphase plate (MP) and polar body (PB) as the sources of genetic material. In the first experiment, GT was established using MP (GT-MP), and a sperm concentration of 1 × 106 or 0.5 × 106 spermatozoa/ml gave similar fertilization rates. The cleavage rate (50%) and blastocyst rate (13.6%) in the GT-MP group was lower than that of the in vitro production control group (80.2% and 32.6%, respectively). The second experiment evaluated the same parameters using PB instead of MP; the GT-PB group had lower fertilization (82.3% vs. 96.2%) and blastocyst (7.7% vs. 36.8%) rates than the control group. No differences in the amount of mitochondrial DNA (mtDNA) were observed between groups. Finally, GT-MP was performed using vitrified oocytes (GT-MPV) as a source of genetic material. The cleavage rate of the GT-MPV group (68.4%) was similar to that of the vitrified oocytes (VIT) control group (70.0%) and to that of the control IVP group (81.25%, P < 0.05). The blastocyst rate of GT-MPV (15.7) did not differ neither from the VIT control group (5.0%) nor from the IVP control group (35.7%). The results suggested that the structures reconstructed by the GT-MPV and GT-PB technique develop in embryos even if vitrified oocytes are used.

Malignancies after renal transplantation: experience of a Mexican referral center.

OBJECTIVE: To report the incidence of cancer after renal transplantation at a referral center in a developing country. MATERIALS AND METHODS: Consecutive patients receiving renal transplantations during a 48-year period at Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran were analyzed. The standardized incidence ratio (SIR) was calculated based on data from GLOBOCAN 2012. RESULTS: From 1257 patients, 98 (8%) developed 143 malignancies. The SIR of all the cohort was 4.1 (95% CI 3.2-5.1). The relative risks of male and female transplant recipients were 4.6 and 3.5 times greater than the risk of cancer of the general population, respectively. The most common malignancy was non-melanoma skin cancer (52%). The malignancy that associated with the greater relative risk was Kaposi sarcoma (SIR: 200), followed by lymphomas (SIR: 30). A multivariate analysis comparing patients with cancer and controls confirmed that receiving a three-drug regimen as final treatment, prolonged immunosuppression, and patients undergoing a second renal transplantation were factors associated with increased cancer development. CONCLUSION: To date, there is paucity of data from developing countries. We reported the results from a National Health Institute in Mexico including a large cohort with a long follow-up, demonstrating differences within frequencies and risks compared to other regions of the world.

Effect of sex on cryotolerance of bovine embryos produced in vitro.

Male and female embryos are known to be different in developmental kinetics, metabolism, gene expression, and epigenetic patterns. Therefore, the objective of this study was to clarify whether the morphological criteria used to select embryos for cryopreservation lead to a deviation in the male:female ratio, and whether vitrification effects vary according to embryo sex. Initially, five sires were tested to evaluate the effect of the bull on embryo development, sex ratio, speed of development, and response to cryopreservation. Results showed that bulls affected (P < 0.05) embryo production, response to cryopreservation, and sex ratio. Then, one bull was selected, and used to produce embryos in vitro to characterize the responses of male and female embryos to vitrification. Results suggested that male and female embryos have the same morphological responses to vitrification, as no differences (P > 0.05) were observed between the two sexes in post-warming survival and re-expansion rates. However, their molecular responses as evaluated by gene expression (FOSL1, HSPB1, CASP3, CASP8, HSPA5, HSPA1A, G6PD, and PGK1) analysis indicated an effect of sex on vitrification; vitrified female embryos exhibited higher mRNA levels of HSPA1A, CASP3, and G6PD compared to their male counterparts. In conclusion, bulls affected embryo production, speed of development, sex ratio, and response to cryopreservation. Male and female embryos differed in their molecular responses to vitrification; and also, deviations in the male:female ratio when selecting embryos for cryopreservation were confirmed.

Correlation of T Cell Subsets and Hypercholesterolemia of the Donor and Its Association with Acute Graft-versus-Host Disease.

Background: Acute graft-versus-host disease (aGVHD) is an important cause of death following allogeneic hematopoietic stem cell transplantation (allo-HSCT). The association between cholesterol and aGVHD was previously described potentially, resulting from pro-inflammatory responses associated with hypercholesterolemia. The aim of this study was to correlate T cell subsets in donor bone marrow (BM) samples with their levels of cholesterol and associate these results with recipients who developed aGVHD and those who did not. Materials and Methods: A prospective study was performed in 39 donor samples. T cell subsets were analyzed by flow cytometry. Results: Eleven (28%) donors had hypercholesterolemia. Donor samples with hypercholesterolemia had less Tregs compared to donors with normal levels of cholesterol (22.69 (IQR=30.6) cells/µL vs 52.62 (IQR=44.68) cells/µL, p=0.04). Among all individuals in the cohort, aGVHD was observed in 21%: 36% from donors with hypercholesterolemia versus 14% from donors with normal levels of cholesterol. Conclusion: As we described the association between hypercholesterolemia and diminished Tregs, our results might suggest that normalizing the levels of total cholesterol in the donor, prior performing allo-HSCT, might be an effective approach to diminish the risk of the receptor to develop aGVHD.

Added Sugar Consumption and Chronic Oral Disease Burden among Adolescents in Brazil.

Chronic oral diseases are rarely studied together, especially with an emphasis on their common risk factors. This study examined the association of added sugar consumption on "chronic oral disease burden" among adolescents, with consideration of obesity and systemic inflammation pathways through structural equation modeling. A cross-sectional study was conducted of a complex random sample of adolescent students enrolled at public schools in São Luís, Brazil ( n = 405). The outcome was chronic oral disease burden, a latent variable based on the presence of probing depth ≥4 mm, bleeding on probing, caries, and clinical consequences of untreated caries. The following hypotheses were tested: 1) caries and periodontal diseases among adolescents are correlated with each other; 2) added sugar consumption and obesity are associated with chronic oral disease burden; and 3) chronic oral disease burden is linked to systemic inflammation. Models were adjusted for socioeconomic status, added sugar consumption, oral hygiene behaviors, obesity, and serum levels of interleukin 6 (IL-6). All estimators of the latent variable chronic oral disease burden involved factor loadings ≥0.5 and P values <0.001, indicating good fit. Added sugar consumption (standardized coefficient [SC] = 0.212, P = 0.005), high IL-6 levels (SC = 0.130, P = 0.036), and low socioeconomic status (SC = -0.279, P = 0.001) were associated with increased chronic oral disease burden values. Obesity was associated with high IL-6 levels (SC = 0.232, P = 0.001). Visible plaque index was correlated with chronic oral disease burden (SC = 0.381, P < 0.001). Our finding that caries and periodontal diseases are associated with each other and with added sugar consumption, obesity, and systemic inflammation reinforces the guidance of the World Health Organization that any approach intended to prevent noncommunicable diseases should be directed toward common risk factors.

Novel bovine-associated pVAPN plasmid type in Rhodococcus equi identified from lymph nodes of slaughtered cattle and lungs of people living with HIV/AIDS.

Rhodococcus equi is a well-recognized Gram-positive intracellular facultative bacterium that is opportunistic in nature, which causes pyogranulomatous infections in humans and multiple host animals. The pathogenicity of the microorganism has been attributed to the presence of plasmid-encoded virulence-associated proteins (Vap). To date, three host-associated virulence plasmid types of R. equi have been identified as follows: the circular pVAPA and pVAPB, related, respectively, to equine and porcine isolates, and a recently described linear pVAPN plasmid associated with bovine strains, although these three types are found in human isolates. Recent phylogenomic studies support the evidence that human R. equi infection is zoonotically acquired. Nevertheless, data regarding distribution and prevalence of the host-adapted virulence plasmid types of R. equi isolated from meat animals are scarce or unnoticed. Here, the three host-associated virulence plasmid types (pVAPA, pVAPB, and pVAPN) were investigated in 154 R. equi isolates recovered from lymph nodes of cattle with lymphadenitis (n = 31), faeces of cattle without enteric signs (n = 49), as well as different clinical specimens from human patients (n = 74). The analysis of virulence profile of 74 R. equi from humans revealed six (8.1%) isolates pVAPB (type 8), two (2.7%) pVAPN, and one (1.3%) pVAPB (type 11), all of which were from lung samples from people living with HIV/AIDS. From the lymph node samples of cattle, 41.9% (13 of 31) isolates revealed pVAPN type, whereas all isolates from faecal samples were negative for three host-associated types. Here, recently described bovine-associated pVAPN type was detected in R. equi isolates recovered from the lungs of people living with HIV/AIDS and lymph nodes from slaughtered cattle intended for human consumption; a finding that represents a public health concern, mainly in countries where undercooked or raw meat are traditionally consumed.

Study of genetic variability in pigs after the traditional breeding program.

Molecular markers are tools used to improve genetic gains. The objective of this study was to analyze the security of alleles of molecular marker genes for characteristics of economic interest in a pure population of pigs. After the extraction of DNA from the hair of 272 Large White matrices, the allele and genotype frequency of single nucleotide polymorphism was performed using the ARMS-PCR Multiplex technique in the DGAT1, LEPR, H-FABP, MC4R, and SREBF1 genes using RFLP-PCR for the GH gene. After capillary electrophoresis in an automated DNA sequencing of the DGAT1, LEPR, H-FABP, and SREBF1 genes, no polymorphisms were found. Only the MC4R marker presented 100% heterozygosity. For the GH gene, 209 of the initial population samples were genotyped. The PCR product (605 bp) was digested with the restriction enzyme DdeI, with fragments being of 335, 148, and 122 bp for the D1 allele and 457 and 148 bp for the D2 allele. The genotypic frequency obtained of D1D2 was 88% and of D2D2 was 22%. The D1 allele presented a frequency of 11% and the D2 allele of 89%. The high intensity of selection for commercial breeds justifies the absence or the low number of polymorphisms for the genes studied.

Breast self-exam and patient interval associate with advanced breast cancer and treatment delay in Mexican women.

PURPOSE: The objective of this study was to compare treatment intervals in breast cancer patients according to the detection method (breast self-exam vs screening). PATIENTS AND METHODS: We conducted a retrospective analysis including 291 breast cancer patients at a Mexican tertiary referral hospital. RESULTS: Breast cancer detection method was mostly breast self-exam (60%). The median patient interval was 60.5 days, and was associated with marital status and socioeconomic level. Differences between the two groups were statistically significant for global interval, p = 0.002; however, health system interval was not statistically different. CONCLUSION: In our country, breast cancer screening is opportunistic, with several weaknesses within its management and quality systems. Our study showed that even in specialized health care centers, breast cancer is detected by self-exam in up to 2/3 of patients, which can explain the advanced stages at diagnosis in our country. In developing countries, the immediate health care access for breast cancer patients should be prioritized as an initial step to reduce the global treatment initiation interval in order to reduce mortality.

Pre-Multidrug-Resistant Mycobacterium tuberculosis Infection Causing Fatal Enteric Disease in a Dog from a Family with History of Human Tuberculosis.

This report describes a fatal case of a pet dog with major enteric signs owned by a family that has experienced cases of pulmonary tuberculosis (TB) in the household. Clinical and epidemiological aspects, imaging data, microbiological, haematological and histopathological examinations were assessed to diagnosis of disease. gyrB-RFLP, spoligotyping and MIRU-VNTR allowed molecular detection of M. tuberculosis strain from S family. The resazurin microtiter assay indicated that all isolates were resistant to isoniazid, ethambutol, ciprofloxacin, ofloxacin, streptomycin and amikacin. The public health concerns related to canine tuberculosis and risk of the dissemination by pets of M. tuberculosis pre-multidrug-resistant (PMD) to isoniazid, ethambutol and other first-line drugs used in human therapy of TB are discussed. We believe this to be the first report of PMD M. tuberculosis infection in a dog presenting mainly enteric manifestation, confirmed as S lineage by molecular methods, owned by a family in which TB has spread in the household for generations.

Short Communication Quick method for identifying horse (Equus caballus) and donkey (Equus asinus) hybrids.

The domestication of the Equus genus 5000-6000 years ago has influenced the history of human civilization. As soon as horse and donkey species had been domesticated, they were crossbred, producing humanity's first documented attempt at animal genome manipulation. Since then, the mule (male donkey x female horse) and the reciprocal cross (the hinny, male horse x female donkey) have been the most common equine hybrids in the world. Due to their hybrid vigor, mules and hinnies have been intensively used for carrying loads and people and for tilling the land. Despite their importance, visual distinction of mules and hinnies is difficult due to high phenotypic resemblance. However, the distinction between these two hybrids is of pivotal importance for equid breeders and ranchers. In this study, an easy, low-cost, effective, and fast multiplex-polymerase chain reaction method was developed to distinguish the maternal origin of mules and hinnies, targeting the hyper-variable mitochondrial DNA D-loop region. This methodology can help breeders, ranchers, animal science professionals, and researchers manage their equine herds with more confidence and precision.

Molecular markers for oocyte competence in bovine cumulus cells.

This study aimed to quantify the expression of candidate genes in cumulus cells (CCs) from cumulus-oocyte complexes (COCs) with high and low potential for in vitro development up to the blastocyst stage. First, the effects of individual culture and biopsy on embryo development were evaluated. Individuals cultured using the well of the well system were compared with individuals cultured in 20 µL droplets (microdroplets) and those cultured in groups (control). Blastocyst rates were lower for the individual culture systems (P < 0.05; well of the well = 17.9%, n = 95; microdrop = 26.3%, n = 95) than for the control group (45.0%, n = 209). Second, the effects of biopsy on embryo production were compared between the control and microdroplet cultures, and no effects (P > 0.05) were observed for either group. Finally, the expression profiles of glypican 4 (GPC4), IGF4-binding protein, follicle-stimulating hormonereceptor, growth hormone receptor, epidermal growth factor receptor, fibroblast growth factor 11, solute carrier family 2 member 1, solute carrier family 2 member 3,sprouty homolog 1, versican, and keratin protein 8 in CCs obtained by biopsy were quantified by real-time polymerase chain reaction. Cumulus cells were categorized on the basis of the fates of the COCs: expanded blastocyst, cleaved and arrested, and uncleaved. The GPC4 gene was overexpressed (P = 0.007) in CCs from oocytes that formed embryos compared with those that produced cleaved and arrested embryos. We concluded that individual culture reduced blastocyst production; however, biopsy did not affect embryo development. The profile of GPC4 expression can be used as a marker to distinguish COCs with potential for embryo development from those with limited developmental potential.

Characterization of allele-specific expression of the X-linked gene MAO-A in trophectoderm cells of bovine embryos produced by somatic cell nuclear transfer.

Somatic cell nuclear transfer (SCNT) may affect epigenetic mechanisms and alter the expression of genes related to embryo development and X chromosome inactivation (XCI). We characterized allele-specific expression of the X-linked gene monoamine oxidase type A (MAO-A) in the trophectoderm (TF) of embryos produced by SCNT. Total RNA was isolated from individual biopsies (N = 25), and the allele-specific expression assessed by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism. Both paternal and maternal alleles were expressed in the trophectoderm. However, a higher frequency of the mono-allelic expression of a specific allele was observed (N = 17; 68%), with the remaining samples showing the presence of mRNA from both alleles (N = 8; 32%). Considering that MAO-A is subject to XCI in bovine, our results suggest that SCNT may influence XCI because neither an imprinted (mono-allelic expression in all samples) nor a random (presence of mRNA from both alleles in all samples) pattern of XCI was observed in TF. Due to the importance of XCI in mammalian embryo development and its sensitivity to in vitro conditions, X-linked genes subject to XCI are candidates for use in the development of embryo quality molecular markers for assisted reproduction.

Ovarian follicular dynamics in 2 to 3 months old Nelore calves (Bos taurus indicus)

This study aimed to evaluate reproductive physiology aspects in 2 to 3 months old Nelore (Bos taurus indicus) calves. Follicular dynamics was monitored daily by ultrasound in ten calves during 18 consecutive days. Calves younger than 2.25 months old (n = 4) had maximum follicle diameters ≤2.5 mm, so only the other animals (n = 6) continued to be monitored by ultrasound. The mean maximum diameter of the ovaries was 13.6 ± 0.6 mm, which had 31.4 ± 3.45 visible follicles. A successive anovulatory follicular wave-like pattern was identified when each wave showed a dominant follicle, including a variable number of other follicles smaller in size (subordinate follicles), during the observational period. Three consecutive follicular growth waves were detected per animal, during this time interval, when 50% of the animals (n = 3) showed two waves and the other half (n = 3) had three waves. The first day of detection of the dominant follicle, retrospectively identified at a diameter of 2 mm, was defined as the day of wave emergence on each wave (day zero). Considering the grouped data, the follicular wave length was 9.2 ± 2.0 days and the correlated dominant follicle began its regression at 6.33 ± 1.63 days after the day of its emergence. The dominant follicles had a growth rate of 0.23 ± 0.06 mm/day and reached the maximum diameter of 3.03 ± 0.17 mm. Despite the smaller ovarian and follicle diameters in 2 to 3 months old Nelore calves (Bos taurus indicus) compared to studies in post pubertal females; the follicular wave-like pattern and the number of recruited follicles were similar to the reported in Zebu females in reproductive activity. These data can characterize an early critical moment on the reproductive development of these animals.(AU)

Ovarian follicular dynamics in 2 to 3 months old Nelore calves (Bos taurus indicus)

This study aimed to evaluate reproductive physiology aspects in 2 to 3 months old Nelore (Bos taurus indicus) calves. Follicular dynamics was monitored daily by ultrasound in ten calves during 18 consecutive days. Calves younger than 2.25 months old (n = 4) had maximum follicle diameters ≤2.5 mm, so only the other animals (n = 6) continued to be monitored by ultrasound. The mean maximum diameter of the ovaries was 13.6 ± 0.6 mm, which had 31.4 ± 3.45 visible follicles. A successive anovulatory follicular wave-like pattern was identified when each wave showed a dominant follicle, including a variable number of other follicles smaller in size (subordinate follicles), during the observational period. Three consecutive follicular growth waves were detected per animal, during this time interval, when 50% of the animals (n = 3) showed two waves and the other half (n = 3) had three waves. The first day of detection of the dominant follicle, retrospectively identified at a diameter of 2 mm, was defined as the day of wave emergence on each wave (day zero). Considering the grouped data, the follicular wave length was 9.2 ± 2.0 days and the correlated dominant follicle began its regression at 6.33 ± 1.63 days after the day of its emergence. The dominant follicles had a growth rate of 0.23 ± 0.06 mm/day and reached the maximum diameter of 3.03 ± 0.17 mm. Despite the smaller ovarian and follicle diameters in 2 to 3 months old Nelore calves (Bos taurus indicus) compared to studies in post pubertal females; the follicular wave-like pattern and the number of recruited follicles were similar to the reported in Zebu females in reproductive activity. These data can characterize an early critical moment on the reproductive development of these animals.

Trueperella pyogenes multispecies infections in domestic animals: a retrospective study of 144 cases (2002 to 2012).

BACKGROUND: Formerly, Arcanobacterium pyogenes was recently renamed Trueperella pyogenes. This opportunistic bacterium is related to miscellaneous pyogenic infections in animals. Most studies involving T. pyogenes are case reports, whereas few surveys have focused the major aspects of T. pyogenes infections involving a case series study design. OBJECTIVE: The aim of this study was to retrospectively evaluate selected epidemiological and clinical aspects, as well as the in vitro antimicrobial susceptibility pattern of 144 cases of T. pyogenes infections among domestic animals from 2002 to 2012. ANIMALS AND METHODS: T. pyogenes was isolated from different clinical specimens from cattle, goats, sheep, pigs, horses, dogs, and buffaloes. Correlations were assessed by the Chi-square or Fisher's exact tests. RESULTS: Mastitis (45.1%), abscesses (18.0%), pneumonia (11.1%), and lymphadenitis (9.0%) were the most common clinical manifestations. In addition, the organism was also isolated from other miscellaneous clinical specimens from cases of septicemia, encephalitis, pyometra, prostatitis, orchitis, seminal vesiculitis, pericarditis, and omphalitis. No statistical association was observed between T. pyogenes infections and age, gender, or season across the study. The most effective drugs against the pathogen were florfenicol (99.1%), cefoperazone (96.0%), cephalexin (95.0%), and ceftiofur (94.8%). High resistance rates were observed against trimethoprim-sulfamethoxazole (49.3%), followed by norfloxacin (10.9%) and tetracycline (9.2%). CONCLUSIONS: This study highlights the diversity of clinical manifestations and the opportunistic behavior of T. pyogenes infections in domestic animals, with predominance of mastitis, abscesses, pneumonia, and lymphadenitis. It also reinforces the importance of knowing the susceptibility profile before initiating therapy, to improve antimicrobial therapy approaches.

Vitrification of bovine oocytes at different meiotic stages using the Cryotop method: assessment of morphological, molecular and functional patterns.

This study aimed to investigate the functional, morphological and molecular patterns of bovine oocytes vitrified at different times during in vitro maturation (IVM). Four groups of oocytes were used: non-vitrified control oocytes (CG), oocytes vitrified at 0 h (V0), oocytes vitrified after 8 h of IVM (V8) and oocytes vitrified after 22 h of IVM (V22). After vitrification, the oocytes were warmed and then returned to the incubator to complete a total of 24h of IVM. To evaluate the effect of vitrification, the nuclear maturation and fertilization rates were assessed by lacmoid staining and ultrastructural electron microscopy. The cleavage and blastocyst rates were evaluated at D2, D7 and D8. The expression levels of CASP3, TP53, HDAC2, SUV39H1 and DNMT1 were investigated by RT-qPCR. The nuclear maturation, oocyte fertilization, cleavage and blastocyst rates were higher (P < 0.05) in the CG group (80%; 81.3%; 88.5%; and 35.8%) than in the V0 (44%; 44.6%; 22.7%; and 2.6%), V8 (50%; 63%; 21.5%; and 2.2%) and V22 (55.5%; 66.9%; 24.1%; and 4.6%) groups. Ultrastructural analysis revealed significant damage within the cytoplasm of all vitrified groups, but more severe degeneration was observed in the V22 group. The gene expression profiles were not affected by vitrification (P > 0.05). In conclusion, cytoplasm degeneration seems to be the most severe form of damage caused by vitrification. The use of the Cryotop method for vitrification severely reduces bovine oocyte viability regardless of whether it is performed at GV, GVBD or MII stage.

Antioxidantes associados à pressão hidrostática sobre a viabilidade embrionária pós-desvitrificação / Association between antioxidant and hidrostatic pressure on the embryonic viability after

Avaliou-se o efeito de dois antioxidantes no cultivo de embriões pós-desvitrificação, associados ounão à pressão hidrostática (PH) em três experimentos. O primeiro avaliou a interação entre PH e antioxidante(β-mercaptoetanol - BME, cisteamina - CYST e BME + CYST). O segundo, similar ao primeiro, vitrificou osembriões uma hora após passarem ou não pela PH. Os parâmetros foram taxas de eclosão e degeneração com24 e 48 h após passar pela PH (experimento 1) e 12, 24, 48 e 72 h pós-desvitrificação (experimento 2). Oexperimento 3 avaliou a taxa de prenhez de embriões cultivados por 12 h com/sem BME. O primeiroexperimento não demonstrou interação entre os tratamentos. No segundo, os resultados foram similares para BX.O tratamento BME + CYST obteve melhor taxa de eclosão dos BL com 48 e 72 h (76,04%). O mesmo fatoocorreu para a taxa de degeneração às 24 h (BME + CYST = 7,29%; 32,29% controle). Ao transferir os embriões(n = 55), percebeu-se uma similaridade em todos os grupos (38,9% CONT; 16,7% VITRIF e 31,6% BME). Opresente trabalho mostrou que o uso da pressão hidrostática e de BME e CYST não influencia as taxas deeclosão, degeneração e de prenhez de embriões vitrificados.

This study aimed to evaluate different antioxidants in embryo culture after vitrification, with or withoutthe previous use of hydrostatic pressure (PH). Three experiments were designed to evaluate the interactionbetween PH and antioxidants (β-mercaptoethanol - BME, cysteamine - CYST and BME + CYST) in fresh andvitrified in vitro produced embryos. In experiment 1 hatching and degeneration rates were evaluated at 24 and48 h after passing through the PH and in experiment 2 the same parameters were evaluated at 12, 24, 48 and72 h after heating. The last step of the study evaluated the pregnancy rate of vitrified embryos, cultured for 12 hwith / without BME. The first experiment showed no difference between treatments. The second found similarresults for all parameters evaluated in BX embryos. Note that the BME + CYST treatment obtained a betterhatching rate in BL with 48 and 72 h (76.04%) than the control group (45.83%). The same behavior wasobserved in degeneration 24 h, where the BME + CYST group was 7.29% against 32.29% in the control.However, the pregnancy rates (55 embryo transfers) were not different between control fresh, control vitrifiedand BME groups (38.9%, 16.7% and 31.6%, respectively). This study showed that the use of hydrostaticpressure and antioxidant had no effect on the evaluated parameters.

Antioxidantes associados à pressão hidrostática sobre a viabilidade embrionária pós-desvitrificação / Association between antioxidant and hidrostatic pressure on the embryonic viability after

Avaliou-se o efeito de dois antioxidantes no cultivo de embriões pós-desvitrificação, associados ounão à pressão hidrostática (PH) em três experimentos. O primeiro avaliou a interação entre PH e antioxidante(β-mercaptoetanol - BME, cisteamina - CYST e BME + CYST). O segundo, similar ao primeiro, vitrificou osembriões uma hora após passarem ou não pela PH. Os parâmetros foram taxas de eclosão e degeneração com24 e 48 h após passar pela PH (experimento 1) e 12, 24, 48 e 72 h pós-desvitrificação (experimento 2). Oexperimento 3 avaliou a taxa de prenhez de embriões cultivados por 12 h com/sem BME. O primeiroexperimento não demonstrou interação entre os tratamentos. No segundo, os resultados foram similares para BX.O tratamento BME + CYST obteve melhor taxa de eclosão dos BL com 48 e 72 h (76,04%). O mesmo fatoocorreu para a taxa de degeneração às 24 h (BME + CYST = 7,29%; 32,29% controle). Ao transferir os embriões(n = 55), percebeu-se uma similaridade em todos os grupos (38,9% CONT; 16,7% VITRIF e 31,6% BME). Opresente trabalho mostrou que o uso da pressão hidrostática e de BME e CYST não influencia as taxas deeclosão, degeneração e de prenhez de embriões vitrificados.(AU)

This study aimed to evaluate different antioxidants in embryo culture after vitrification, with or withoutthe previous use of hydrostatic pressure (PH). Three experiments were designed to evaluate the interactionbetween PH and antioxidants (β-mercaptoethanol - BME, cysteamine - CYST and BME + CYST) in fresh andvitrified in vitro produced embryos. In experiment 1 hatching and degeneration rates were evaluated at 24 and48 h after passing through the PH and in experiment 2 the same parameters were evaluated at 12, 24, 48 and72 h after heating. The last step of the study evaluated the pregnancy rate of vitrified embryos, cultured for 12 hwith / without BME. The first experiment showed no difference between treatments. The second found similarresults for all parameters evaluated in BX embryos. Note that the BME + CYST treatment obtained a betterhatching rate in BL with 48 and 72 h (76.04%) than the control group (45.83%). The same behavior wasobserved in degeneration 24 h, where the BME + CYST group was 7.29% against 32.29% in the control.However, the pregnancy rates (55 embryo transfers) were not different between control fresh, control vitrifiedand BME groups (38.9%, 16.7% and 31.6%, respectively). This study showed that the use of hydrostaticpressure and antioxidant had no effect on the evaluated parameters.(AU)