Estrus synchonization with prostaglandin F2α compared to progestogen treatment associated with equine chorionic gonadotropin (eCG) in Santa Inês breed ewes reared in Federal District, Brazil

Este estudo teve por objetivo comparar o uso de um análogo da PGF2α à associação de progestágeno (MAP) com gonadotrofina coriônica equina (eCG) na sincronização do estro em ovelhas da raça Santa Inês. Foram utilizadas 38 fêmeas ovinas submetidas a dois protocolos de sincronização de estro, com os seguintes protocolos: PGF2α (duas doses de 0,53 mg de D-cloprostenol com nove dias de intervalo) e protocolo MAP+eCG (pessário intravaginal com 50 mg de acetato de medroxiprogesterona por doze dias e no momento da remoção do dispositivo aplicação de 250 UI de eCG, IM). Submeteram-se as ovelhas aos diferentes protocolos, com intervalo de dois ciclos estrais. Procedeu-se a uma ultrassonografia transretal no último dia do protocolo, para avaliação do diâmetro do maior e do segundo maior folículo, e à coleta de sangue no dia sete do ciclo estral, para avaliação da concentração sérica de P4. Exame laparoscópico foi realizado no dia 11, após o fim dos protocolos, para contagem de corpos lúteos. Para os parâmetros taxa de sincronização, diâmetro do maior e do segundo maior folículo, período do final do protocolo ao estro e taxa de ovulação, não se observaram diferenças entre os mesmos. Foi observado que o protocolo MAP+eCG produziu concentrações séricas de P4 maiores do que o protocolo PGF2α (3,9 e 2,8 ng/ mL, respectivamente, P<0,05). Nas condições do presente estudo, embora o protocolo MAP+eCG tenha apresentado superioridade em relação à concentração sérica de P4, o protocolo PGF2α foi tão eficiente quanto aquele em sincronizar o estro. __________________________________________________________________________________________ ABSTRACTThe aim of this study was to compare two protocols of estrus synchronization in Santa Inês ewes. Thirty-eight ewes were randomly divided into two groups of estrus synchronization: protocol PGF2α (two doses of 0.530 mg of PGF2α, nine days apart) and protocol MAP+eCG (intravaginal sponge impregnated with medroxyprogesterone acetate, for 12 days, and then an injection of 250 IU of eCG). The experiment was in a cross-over design, two estrous cycles apart. On the final day of protocol, a transrectal ultrasound examination was carried out to measure the size of the largest and second largest ovarian follicles and on day 7 of estrous cycle blood was collected to measure serum P4 concentration. Laparoscopy was carried out on day 11 after the end of protocols to count corpora lutea. Synchronization rate, size of largest and second largest ovarian follicles, interval between the end of the protocol to estrus and ovulation rate did not differ between protocols. Ewes synchronized with MAP+eCG had greater serum P4 concentrations than ewes synchronized with PGF2α (3.9 and 2.8 ng/mL, respectively, P<0.05). Based on the results, it may be concluded that, although the protocol MAP+eCG was superior in inducing higher serum concentration of P4, the protocol PGF2α was equivalent regarding estrus synchronization

Embryo production in Morada Nova and Somalis Brasileira ewes

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR(r)) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR(r) removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes

Composição de fosfolipídios e resistência à vitrificação de blastocistos produzidos in vivo de uma raça de suíno naturalizada brasileira

Blastocistos de suínos foram submetidos ao MALDI-TOF para se identificarem os principais fosfolipídios (PL). Depois, parte destes embriões (D6) foram vitrificados (n=52), ou permaneceram frescos (grupo controle, n=42). Após o aquecimento, os blastocistos foram cultivados in vitro para se avaliar a reexpansão e a eclosão (BE) às 24 e 48 horas. Finalmente, às 48 horas, os BE foram submetidos ao RT-qPCR em busca dos genes BCL2A1, BAK, BAX e CASP3. No MALDI-TOF, a intensidade do íon foi expressa em unidades arbitrárias. O desenvolvimento embrionário foi comparado por qui-quadrado (P0,05) entre blastocistos frescos ou vitrificados às 24 (33,3%, 32,7%) e 48 horas (2,4%, 13,5%). As taxas de eclosão foram maiores (P 0.05) between fresh or vitrified blastocysts at 24 (33.3%; 32.7%) or 48 hours (2.4%; 13.5%). Hatching rates were higher (P< 0.05) for fresh compared to vitrified at 24 (66.7%; 15.4%) and 48 hours (97.6%; 36.0%). BAX was overexpressed (P< 0.05) after vitrification. In conclusion, Piau blastocysts can be cryopreserved by Cryotop. This study also demonstrated that the apoptotic pathway may be responsible for the low efficiency of porcine embryo cryopreservation

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Reproductive caractherization of Santa Inês ewes with mutation on gene GDF-9

Tese (doutorado)–Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2010.No primeiro experimento foi encontrado um novo alelo do GDF-9, nomeado FecGE (Embrapa), que leva a uma substituição da fenilalanina por uma cisteína em uma posição conservadora do peptídeo maduro do gene. Para investigar a associação entre os genótipos FecGE e a taxa de ovulação, 39 ovelhas (15 FecG+/+, 15 FecG+/E e 9 FecGE/E) foram selecionadas dos rebanhos genotipados, e submetidas a dois protocolos de sincronização de estro (PGF2? e MAP + eCG). A taxa de ovulação foi maior (P 0,05) entre o protocolo Dia 0 (CL totais 9,8 ± 5,3; estruturas totais 4,5 ± 4,6; estruturas viáveis 1,6 ± 2,0) e o protocolo tradicional (CL totais 10,0 ± 6,0; estruturas totais 3,5 ± 4,3; estruturas viáveis 1,7 ± 2,4). Para quantidade de corpos lúteos nos ovários (E/E 9,0±6,3; +/E 10,1 ±5,3; +/+ 10,5±5,3), estruturas totais coletadas (E/E 4,9±5,0; +/E 3,1±3,1; +/+ 4,1±5,2), e viáveis (E/E 1,9±2,1; +/E 2,2±2,6; +/+ 0,9±1,7), não houve diferença (P>0,05) entre os diferentes genótipos. Foi observado maior número de animais respondendo ao tratamento superovulatório quando utilizado o protocolo Dia 0 (10/18) em comparação ao protocolo tradicional (6/18). Quando se comparou a quantidade de ovelhas que responderam aos protocolos de superovulação, pelo genótipo não houve diferença significativa: E/E 6/12; +/E 5/12 e +/+ 5/12. Não foi observada alteração nos ovários, no crescimento folicular e nem na produção de embriões viáveis quando se comparou os genótipos. Os dois protocolos foram eficientes em estimular a superovulação e produção de embriões, mas não houve diferença entre os genótipos avaliados quanto a taxa de ovulação. _______________________________________________________________________________ ABSTRACTIn the first experiment was found a new allele of GDF-9, named FecGE (Embrapa), which leads to a substitution of a phenylalanine by a cysteine in a conservative position of the mature peptide in the gene. To investigate the association between the FecGE genotypes (E/E, +/E, and +/+) and the ovulation rate, 39 ewes (15 FecG+/+, 15 FecG+/E and 9 FecGE/E) were selected from the genotyped flocks, and submitted to two estrus synchronization protocol (PGF2α and MPA + eCG). The ovulation rate was greater (P<0.001) in the homozygote mutant (E/E) group which showed an 82% increase in CL average (2.22± 0.12), as well as the highest frequency (96.3%) of multiple-ovulating ewes, when compared to +/E and +/+ groups. The heterozygote group (+/E) presented no difference (P=0.612) in CL average (1.34±0.08), or in the frequency (31.8%) of ewes with multiple ovulations, when compared to the wild-type ewes (1.22±0.11 and 14.6%, respectively). Was observed a genotype effect on the number of twins per ewe (P=0.0136); E/E ewes showed 44% of twinpregnancy, +/E 14% e +/+ 0%. In conclusion, the homozygote ewes had higher ovulation rate and prolificacy than heterozygote and the wild-types. In second experiment the superovulation and embryo recovery was made in 18 ewes (6 +/+, 6 +/E e 6 E/E) to compare ovarian response among genotype. Two protocols of superovulation were used: traditional protocol and Day 0 protocol. Was used fresh semen for AI of one male with proved fertility. Five days after AI the embryos were recovered cirurgically, by laparotomy. There was no difference among evaluated parameters (P>0.05) between Day 0 protocol (total CL 9.8 ± 5.3; total structures 4.5 ± 4.6; viable structures 1.6 ± 2.0) and traditional protocol (total CL 10.0 ± 6.0; total structures 3.5 ± 4.3; viable structures 1.7 ± 2.4). Corpora lutea in the ovaries (E/E 9.0±6.3; +/E 10.1 ±5.3; +/+ 10.5±5.3), total structure recovered (E/E 4.9±5.0; +/E 3.1±3.1; +/+ 4.1±5.2), and viable embryos (E/E 1.9±2.1; +/E 2.2±2.6; +/+ 0.9±1.7), there was no difference (P>0.05) among groups of genotype animals. Was observed higher number of animals answering the superovulation treatment when used Day 0 protocol (10/18) in comparison with traditional protocol (6/18). The ewes that answered to superovulation protocol, by genotype, there were no significative difference E/E 6/12; +/E 5/12 e +/+ 5/12. Was not observed alteration in ovaries, in follicular growth and neither in viable embryo production when compare among genotypes. Both protocols were efficient in superovulation and embryo production, but there were no difference among evaluated genotypes in ovulation rate

Sincronização de estro com prostaglandina F2 versus progesterona associada à gonadotrofina coriônica eqüina (eCG) em ovelhas deslanadas no Distrito Federal

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2008.Este estudo teve por objetivo comparar o uso de PGF2 à associação de progesterona (P4) com gonadotrofina coriônica eqüina (eCG) na sincronização do estro em ovelhas da raça Santa Inês. Foram utilizadas 38 fêmeas ovinas divididas em dois grupos aleatoriamente e submetidas a dois protocolos de sincronização de estro: protocolo PGF2 (duas doses de 0,530 mg de PGF2 com nove dias de intervalo) e protocolo P4+eCG (pessário intravaginal de acetato de medroxiprogesterona por 12 dias e no momento da remoção do dispositivo aplicação de 250 UI de eCG). As ovelhas foram submetidas ao “cross-over”, com intervalo de dois ciclos estrais. Procedeu-se uma ultra-sonografia transretal no último dia do protocolo para avaliar diâmetro do maior e do segundo maior folículo e foi coletado sangue no dia sete do ciclo estral para avaliação da concentração sérica de P4. Exame laparoscópico foi realizado no dia 11 após o fim dos protocolos para contagem de corpos lúteos. Para os parâmetros taxa de sincronização, diâmetro do maior e do segundo maior folículo, período do final do protocolo ao estro e taxa de ovulação não foram observadas diferenças entre os protocolos. Foi observado que o protocolo P4+eCG produziu concentrações séricas de P4 maiores do que o protocolo PGF2 (3,9 e 2,8 ng/mL, respectivamente, P<0,05). Nas condições do presente estudo, embora o protocolo P4+eCG tenha apresentado superioridade em relação à concentração sérica de P4, o protocolo PGF2 foi tão eficiente em sincronizar o estro quanto o P4+eCG. _______________________________________________________________________________ ABSTRACTThe aim of this study was to compare two protocols of estrus synchronization in Santa Inês ewes. Thirty-eight ewes were randomly divided into two groups and treated with two protocols of estrus synchronization: protocol PGF2 (two doses of 0.530 mg of PGF2, nine days apart) and protocol P4+eCG (intravaginal sponge impregnated with medroxyprogesterone acetate, for 12 days and then an injection of 250 IU of eCG). The experiment was in a cross-over design, two estrous cycles apart. On the final day of protocol a transrectal ultrasound examination was carried out to measure the size of the largest and second largest ovarian follicles and on day 7 of estrous cycle blood was collected to measure serum P4 concentration. Laparoscopy was carried out on day 11 after the end of protocols to count corpora lutea. Synchronization rate, size of largest and second largest ovarian follicles, hours between the end of the protocol to estrus and ovulation rate did not differ between protocols. Ewes synchronized with P4+eCG had greater serum P4 concentrations than ewes synchronized with PGF2 (3.9 and 2.8 ng/mL, respectively, P<0.05). Based on the results, it may be concluded that although the protocol P4+eCG was superior in inducing higher serum concentration of P4, the protocol PGF2 was equivalent regarding estrus synchronization

SINCRONIZAÇÃO DE ESTRO COM PROSTAGLANDINA F2α versus PROGESTÁGENO ASSOCIADO À GONADOTROFINA CORIÔNICA EQUINA (eCG) EM OVELHAS SANTA INÊS NO DISTRITO FEDERAL, BRASIL

The aim of this study was to compare two protocols ofestrus synchronization in Santa Inês ewes. Thirty-eight ewes wererandomly divided into two groups of estrus synchronization: protocolPGF2α (two doses of 0.530 mg of PGF2α, nine days apart)and protocol MAP+eCG (intravaginal sponge impregnated withmedroxyprogesterone acetate, for 12 days, and then an injectionof 250 IU of eCG). The experiment was in a cross-over design,two estrous cycles apart. On the final day of protocol, a transrectalultrasound examination was carried out to measure the size of thelargest and second largest ovarian follicles and on day 7 of estrouscycle blood was collected to measure serum P4 concentration.Laparoscopy was carried out on day 11 after the end of protocols to count corpora lutea. Synchronization rate, size of largest andsecond largest ovarian follicles, interval between the end of theprotocol to estrus and ovulation rate did not differ between protocols.Ewes synchronized with MAP+eCG had greater serumP4 concentrations than ewes synchronized with PGF2α (3.9 and2.8 ng/mL, respectively, P<0.05). Based on the results, it may beconcluded that, although the protocol MAP+eCG was superior ininducing higher serum concentration of P4, the protocol PGF2αwas equivalent regarding estrus synchronization

Blastocyst development of oocytes recovered from slaughterhouse ovaries (CONT) and by ovum pick-up (OPU) from the ovaries of non-superstimulated females (IMA), superstimulated females (FSH) and superstimulated females that received an ovulation inducer (MII) that were vitrified (VIT) at the metaphase II stage.

<p>^a,b,c,d Values with different superscripts in the same column are different at P<0.05.</p><p>* Hatched blastocyst at D8 as a percentage of oocyte number.</p><p>Blastocyst development of oocytes recovered from slaughterhouse ovaries (CONT) and by ovum pick-up (OPU) from the ovaries of non-superstimulated females (IMA), superstimulated females (FSH) and superstimulated females that received an ovulation inducer (MII) that were vitrified (VIT) at the metaphase II stage.</p

Experimental design flow diagram.

<p>Flow diagram of experimental design for the different treatments. Oocytes recovered from slaughterhouse ovaries (CONT), obtained by OPU from non-superstimulated females (IMA) and from superstimulated females (FSH) were matured in vitro. In vivo-matured oocytes were obtained by OPU from superstimulated females that received an ovulation inducer 24 hours previously (MII). A sample of matured oocytes from each of four groups was used to study the composition of plasma membrane phospholipids using MALDI-TOF. The remaining oocytes were divided in half, one half consisting of non-vitrified fresh oocytes (CONT, IMA, FSH and MII) and other of vitrified/ warmed oocytes (CONT Vit, IMA Vit, FSH Vit and MII Vit). At the end of the warming process, the eight groups were used for in vitro fertilization and culture. Cleavage at D2 and blastocyst development at D7 and D8 were evaluated. At D8, all of the blastocysts were measured, and those larger than 160 μM in diameter were stained for total cell number counting.</p

Comparison of the relative intensity of most dispersed ions after mass spectrometry (MALDI-TOF) analyses, of oocytes from different maturation systems.

<p>The 760.6 ion corresponds to [PC (34:1) + H] ⁺ and 782.6 to [PC (34:6) + H] ⁺ or [PC (34:1) + Na], both phosphatidylcholines (PC).</p><p>^ab Different letters in the same column indicates statically differences among the treatments after ANOVA according to Tukey’s test (P<0.05).</p><p>CONT = oocytes from slaughterhouse ovaries and matured in vitro; IMA = OPU oocytes from non-stimulated animals and matured in vitro; FSH = OPU oocytes from FSH simulated animal and matured in vitro; MII = OPU oocytes after in vivo maturation.</p><p>The data are expressed as arbitrary unit intensity and standard deviation (±SD), and each ion represents a different phospholipid.</p