Effect of recombinant gonadotropins on embryo quality in superovulated rabbit does and immune response after repeated treatments.

This study aimed first to evaluate the effect of recombinant human FSH (rhFSH) with and without recombinant human LH (rhLH) on fresh and frozen-thawed embryo development and also to analyze the immune response of rabbit does (Oryctolagus cuniculus) subjected to repeated rhFSH treatments. Nulliparous New Zealand White does were used. In Experiment 1, 120 does were superovulated with 25 IU rhFSH alone or in combination with 5% or 10% rhLH (1.25 IU or 2.50 IU rhLH). A total of 1116 embryos at the compacted morula stage were cultured at 38.5 degrees C, 5% CO(2), and saturated humidity for 48 h. The embryo development to hatching blastocyst was significantly lower for the group with 10% rhLH versus that of the control group (65.6 vs. 79.5 for rhFSH+10% rhLH vs. control, respectively). However, no significant difference was found in development to hatching blastocyst for the control, rhFSH alone, and rhFSH+5% rhLH groups. The developmental potential of frozen-thawed embryos obtained from all groups was similar, with an 83.5% in vitro development rate until the expanded blastocyst stage. To detect anti-FSH antibodies, in Experiment 2, does were subject to four superovulation treatments. The hormone administration had a significant effect on immune response in the superovulation group after two treatments (0.14+/-0.074 and 0.15+/-0.076 vs. 0.46+/-0.078 and 0.50+/-0.078 optical density for the first, second, third, and forth cycles, respectively). Nevertheless, none of the treated does had an immune response in both the first and second treatments; on the contrary, a significant increase in the antibody levels was observed in these females at the moment of the third and fourth superovulation treatments. In conclusion, rhFSH superovulation treatments increase the reproductive potential of rabbit does.

Morphometry characterisation of European eel spermatozoa with computer-assisted spermatozoa analysis and scanning electron microscopy.

The aim of the present study was to characterise European eel spermatozoa morphometrically, as a basis for future studies on the morphological effects of methods for sperm cryopreservation and sperm quality. This characterisation was carried out measuring several spermatozoa morphology parameters (head length, width, area and perimeter) by scanning electron microscopy (SEM), in comparison with measurements developed in European eel spermatozoa with computer-assisted morphology analysis (ASMA). Spermatozoa head morphology showed differences in width (1.15+/-0.01 microm versus 1.12+/-0.01 microm), perimeter (14.68+/-0.13 microm versus 13.72+/-0.19 microm) and area (5.36+/-0.06 microm2 versus 1.12+/-0.01 microm2) for ASMA and SEM, respectively. When head length was evaluated, significant differences were found, being higher for SEM methodology (5.09+/-0.04 microm versus 4.29+/-0.03 microm). The curved and elongated spermatozoa head in eels means a problem for the ASMA system (Sperm Class Analyser), Morfo Version 1.1, Imagesp, Barcelona, Spain), causing an error in the length measurements. However, similar results were obtained by both techniques when spermatozoa head length was considered as the greater length between two points within the object (4.29+/-0.03 microm versus 4.31+/-0.04 microm for ASMA and SEM, respectively). In conclusion, this is one of the first applications of ASMA in fish and the first in this species, and confirms this system as a useful tool with wide applications in future fish spermatozoa studies. Width, perimeter and area could be used as parameters for the spermatozoa morphology evaluation, whereas the length requires a new programming of the Imagesp software.