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We have investigated the changes in the mechanical properties of the zona pellucida (ZP), a multilayer glycoprotein coat that surrounds mammalian eggs, that occur after the maturation and fertilization process of the bovine oocyte by using atomic force spectroscopy. The response of the ZP to mechanical stress has been recovered according to a modified Hertz model. ZP of immature oocytes shows a pure elastic behavior. However, for ZPs of matured and fertilized oocyte, a transition from a purely elastic behavior, which occurs when low stress forces are applied, towards a plastic behavior has been observed. The high critical force necessary to induce deformations, which supports the noncovalent long interaction lifetimes of polymers, increases after the cortical reaction. Atomic force microscopy (AFM) images show that oocyte ZP surface appears to be composed mainly of a dense, random meshwork of nonuniformly arranged fibril bundles. More wrinkled surface characterizes matured oocytes compared with immature and fertilized oocytes. From a mechanical point of view, the transition of the matured ZP membrane toward fertilized ZP, through the hardening process, consists of the recovery of the elasticity of the immature ZP while maintaining a plastic transition that, however, occurs with a much higher force compared with that required in matured ZP.

BackgroundProper calving management of dairy herds is a crucial aspect of the bovine life cycle, as it has profound effects on calf viability and on the post-partum course of the dam. The objectives of this study were to monitor the calving process through the use of a remote alarm system and to determine the impact of prompt emergency obstetric procedures in case of dystocia for the prevention of stillbirths and post-partum reproductive pathologies, and for improving herd fertility. Six groups of experimental animals were studied: monitored heifers (n = 60) and multiparous cows (n = 60) were compared with non-observed animals (n = 60 heifers and n = 60 multiparous) giving birth during the same time period and housed in the calving barn, and with unmonitored animals placed in a dry zone (n = 240 heifers and n = 112 multiparous cows).ResultsThe incidence of dystocia ranged from a minimum of 23.4% (monitored multiparous cows) to a maximum of 33.3% (monitored heifers), and there were no differences compared with control groups. However, the rate of stillbirth was higher in control groups than in the monitored groups (P < 0.01). Among both heifers and multiparous cows, the incidence of post-partum uterine infections was higher in the unmonitored animals both in the calving barn (P < 0.01) and in the dry zone (P < 0.05) compared with monitored animals. Among both heifers and multiparous cows, the control groups showed higher rates of foetal membrane retention than did the monitored groups (P < 0.001). The calving-to-conception interval was shorter; in particular, observed heifers showed a significant advantage of approximately 46 days compared with the unmonitored group (P < 0.001) and 32 days compared with the group in the calving barn (P < 0.05). Multiparous cows also had a reduction in the number of days open.ConclusionsThe remote alarm system used to monitor the calving process assured the prompt presence of personnel, improving both the cow’s reproductive efficiency and neonatal viability.

The zona pellucida (ZP) is an extracellular membrane surrounding mammalian oocytes. The so-called zona hardening plays a key role in fertilization process, as it blocks polyspermy, which may also be caused by an increase in the mechanical stiffness of the ZP membrane. However, structural reorganization mechanisms leading to ZP's biomechanical hardening are not fully understood yet. Furthermore, a correct estimate of the elastic properties of the ZP is still lacking. Therefore, the aim of the present study was to investigate the biomechanical behaviour of ZP membranes extracted from mature and fertilized bovine oocytes to better understand the mechanisms involved in the structural reorganization of the ZP that may lead to the biomechanical hardening of the ZP. For that purpose, a hybrid procedure is developed by combining atomic force microscopy nanoindentation measurements, nonlinear finite element analysis and nonlinear optimization. The proposed approach allows us to determine the biomechanical properties of the ZP more realistically than the classical analysis based on Hertz's contact theory, as it accounts for the nonlinearity of finite indentation process, hyperelastic behaviour and material heterogeneity. Experimental results show the presence of significant biomechanical hardening induced by the fertilization process. By comparing various hyperelastic constitutive models, it is found that the Arruda -Boyce eight-chain model best describes the biomechanical response of the ZP. Fertilization leads to an increase in the degree of heterogeneity of membrane elastic properties. The Young modulus changes sharply within a superficial layer whose thickness is related to the characteristic distance between cross-links in the ZP filamentous network. These findings support the hypothesis that biomechanical hardening of bovine ZP is caused by an increase in the number of inter-filaments cross-links whose density should be higher in the ZP inner side.

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