Individual assessment of bovine embryo development using a homemade chamber reveals kinetic patterns of success and failure to reach blastocyst stage.

Most early developmental data are lost in bovine embryo culture systems. We developed and validated a method for culture of bovine embryos in groups that allow individual assessment. An autoclavable low-cost multiembryo chamber (MEC) was prepared using a polyester mesh fixed to a glass coverslip. Embryonic development was not affected by MEC. Compared to conventional bovine culture system (oil-covered drops, control), cleavage (C, 71.2 ± 7.8%; MEC, 74.3 ± 6.0%), blastocyst rate (C, 29.9 ± 4.4%; MEC, 28.3 ± 5.0%) and blastocyst cell number (C, 94.1 ± 9.7; MEC, 92.9 ± 5.3) were similar. Caspase 3 positive cell index in blastocysts was increased in MEC group, but apoptosis rate was below 5% (C, 2.9 ± 0.5; MEC, 4.6 ± 0.6). Using MEC, we performed a retrospective analysis for 'failure' and 'success' embryos, based on their ability to reach the blastocyst stage. We detected the majority of 'success' embryos displayed 8 cells at 48 h post-insemination (hpi) (48.7%), but blastocysts derived from this pattern presented lower cell numbers (91.3 ± 4.2 vs. 107.9 ± 4.9) and higher apoptosis index (6.2 ± 0.6 vs. 4.4 ± 0.5) than blastocysts from 4-cell embryos at 48 hpi. Most (72.0%) embryos that were at morula stage 120 hpi reached blastocyst stage at 168 hpi. Those blastocysts presented more number of cells than blastocysts derived from embryos exhibiting 16 cells at 120 hpi (108.6 ± 4.1 vs. 83.9 ± 4.8). Combination of embryo kinetics data at 48 and 120 hpi revealed high chances of blastocyst formation for patterns: 8 cells/morula, 4 cells/morula, 8 cells/16 cells and 4 cells/16 cells. Blastocysts formed from 4-cell/morula and 8-cell/morula patterns represented 69% of all 168 hpi blastocysts. Blastocysts derived from 4 cells/16 cells displayed decreased apoptosis (3.1 ± 0.6). Our results suggest that MEC can be used for bovine embryo culture without detrimental effects on development and can help to predict blastocyst formation and quality of in vitro fertilization (IVF) embryos. Abbreviations: BSA: bovine serum albumine; COC: cumulus-oocyte complex; FERT-TALP: Tyrode's albumin lactate pyruvate fertilization; FBS: fetal bovine serum; IVF: in vitro fertilization; MEC: multiembryo chamber; PBS: phosphate buffered saline; SOF-AA: synthetic oviductal fluid with amino acids medium; TCM: Tissue Culture Medium.

Breeding of White-tailed Tropicbirds (Phaethon lepturus) in the western South Atlantic / Biologia reprodutiva, tamanho populacional e conservação do rabo-de-palha-de-bico-laranja no Atlântico Sul

Abstract Basic information on natural history is crucial for assessing the viability of populations, but is often lacking for many species of conservation concern. One such species is the White-tailed Tropicbird, Phaethon lepturus (Mathews, 1915). Here, we address this shortfall by providing detailed information on reproductive biology, distribution and threats on the Fernando de Noronha archipelago, Brazil – the largest colony of P. lepturus in the South Atlantic. We assessed reproduction from August 2011 to January 2012 by monitoring tropicbird nests and their contents. A population estimate was obtained through a combination of active searches for nests and by census at sea between 2010 and 2012. Breeding success was calculated by traditional methods. The growth curve of chicks and life table were also calculated. Additional information on nest and mate fidelity and on age of breeding birds was obtained from the banded birds. Our results indicate that the unusual nest form (limestone pinnacles) and predation by crabs may be responsible for the observed patterns of hatching and fledging success. Although the Fernando de Noronha population appears to be stable (at between 100-300 birds), a long term monitoring program would be desirable to assess fluctuations in this globally important population. Conservation strategies should focus on controlling predation by land crabs and tegu lizards.

Resumo Informações básicas sobre história natural são cruciais para acessar a viabilidade de populações, mas são ausentes para muitas espécies que necessitam de conservação. Uma destas espécies é o rabo-de-palha-de-bico-laranja, Phaethon lepturus Daudin, 1802. Aqui, vamos abordar o déficit de dados para esta espécie, fornecendo informações detalhadas sobre a biologia reprodutiva, tamanho da população, distribuição e ameaças em Fernando de Noronha, Brasil – a maior colônia de P. lepturus no Atlântico Sul. Acompanhamos a reprodução do rabo-de-palha-de-bico-laranja de Agosto de 2010 a Janeiro de 2011 monitorando ninhos e seus conteúdos. A estimativa da população foi obtida através de uma combinação de busca ativa de ninhos e censo no mar entre 2010 e 2012. O sucesso reprodutivo foi avaliado por métodos tradicionais. A curva de crescimento da coorte e a tabela de vida também foram obtidas. Além disso, informações sobre fidelidade ao ninho e parceiro e, a idade de reprodutores foi obtida a partir das aves anilhadas anteriormente. Nossos resultados indicam que a forma incomum de ninho (pináculos de calcário) e a predação por caranguejos podem ser responsáveis pelo sucesso observado de eclosão e recrutamento. A população de Fernando de Noronha parece estar estável entre 100-300 aves. No entanto, um programa de monitoramento a longo prazo seria desejável para avaliar as flutuações desta população globalmente importante. As estratégias de conservação devem se concentrar em controlar a predação por caranguejos e lagartos teiú.

Breeding of White-tailed Tropicbirds (Phaethon lepturus) in the western South Atlantic.

Basic information on natural history is crucial for assessing the viability of populations, but is often lacking for many species of conservation concern. One such species is the White-tailed Tropicbird, Phaethon lepturus (Mathews, 1915). Here, we address this shortfall by providing detailed information on reproductive biology, distribution and threats on the Fernando de Noronha archipelago, Brazil - the largest colony of P. lepturus in the South Atlantic. We assessed reproduction from August 2011 to January 2012 by monitoring tropicbird nests and their contents. A population estimate was obtained through a combination of active searches for nests and by census at sea between 2010 and 2012. Breeding success was calculated by traditional methods. The growth curve of chicks and life table were also calculated. Additional information on nest and mate fidelity and on age of breeding birds was obtained from the banded birds. Our results indicate that the unusual nest form (limestone pinnacles) and predation by crabs may be responsible for the observed patterns of hatching and fledging success. Although the Fernando de Noronha population appears to be stable (at between 100-300 birds), a long term monitoring program would be desirable to assess fluctuations in this globally important population. Conservation strategies should focus on controlling predation by land crabs and tegu lizards.