Pregnancy after percutaneous epididymal sperm aspiration in an 81-year-old man with obstructive azoospermia.

There has been observed a trend to delay childbearing, reflecting couple's desire to have children at older ages. Maternal age is a well-known factor that influences the achievement of a pregnancy. In contrast, there are few studies examining the effect of paternal age on the outcomes of assisted reproductive technologies (ART), and results are conflicting. Our patient was vasectomised at the age of 60 years, and his wife was 38 years old. A total of four metaphase II oocytes were inseminated 4 h later (day 0) by intracytoplasmic sperm injection using spermatozoa from percutaneous epididymal sperm aspiration. On day 2, the three resulting embryos were transferred (two had four cells, and one had two cells). Two healthy infants were born at the 37th gestational week by caesarean section. Although the effect of paternal age on ART outcomes and results are conflicting, this case report contributes to point out that advanced age of the man has probably lesser negative influence in the reproductive function.

Birth of healthy twins after intracytoplasmic sperm injection using ejaculated immotile spermatozoa from a patient with Kartagener's syndrome.

This case report demonstrates a successful pregnancy after ICSI combined with hypo-osmotic swelling test in a couple with Kartagener's syndrome with complete immotile ejaculated spermatozoa. Our result suggests that even for complete immotile spermatozoa, the use of hypo-osmotic swelling test is a good alternative to identify viable spermatozoa. When associated with ICSI, it can be a valuable tool to get fertilisation and pregnancy.

Gene banking of the neotropical fish Leporinus obtusidens (Valenciennes, 1836): a protocol to freeze its sperm in the field.

A practical sperm cryopreservation protocol using a dry-shipper and a diluent of simple composition is described for the neotropical fish Leporinus obtusidens (Valenciennes, 1836). The cooling rate of the dry-shipper and its period of useful time, established under laboratory conditions, were respectively 25.7-30.8 degrees C/min (between 0 and -60 degrees C) and 9 days after charging. Sperm donors were selected on the basis of their hyperemic genital papilla and the ability to ooze milt under gentle manual pressure, during the reproductive months of November to January. Milt volume (1.3+/-0.3 mL; n=9 fish), fresh sperm motility rate (93.3+/-2.5%; n=6 fish), and sperm concentration (10.9+/-3.0 x 10(9)spermatozoa/mL of milt) were obtained. The sperm cryopreservation experiments were conducted with the following cryoprotectants (all at 10%, before mixing with milt): dimethyl sulphoxide (DMSO; n=10 fish), methanol (n=6 fish), propanediol (n=6 fish) and ethylene glycol (n=5 fish). Glucose (5%) and hen's egg yolk (10%) made up the diluents containing DMSO, ethylene glycol or propanediol. Milk powder (10%) replaced hen's egg yolk in the diluent containing methanol. Distilled water (up to 100%) completed the diluent solutions. Milt freezing (in 0.5-mL straws) was performed in the dry-shipper after 1:5 (milt:diluent) dilution. Thawed sperm cryopreserved in DMSO-containing diluent and activated by 119 mM NaHCO(3) gave the highest motility rate (62+/-14%). The fertilizing capacity of L. obtusidens sperm was tested using the combination of DMSO-containing diluent as the cryoprotectant and 119 mM NaHCO(3) as the activating solution. Oocytes were obtained from artificial spawning and fertilized with different proportions of spermatozoa. The greatest rate of fertilization (74%) occurred when the ratio of about 112,000 motile spermatozoa:oocyte was used. Thus, a protocol to freeze L. obtusidens sperm can be elaborated as follows. Milt (<1.5 mL fish(-1)) was readily available only in November to January; a simple solution, composed of 10% DMSO (concentration before adding milt), 5% glucose, and 10% hen yolk egg, in distilled water, was used as sperm diluent; cooling rate of 25-30 degrees C/min, yielded in a portable dry-shipper, was adequate to freeze diluted milt (1:5; milt:diluent), in 5-mL straws; about 112,000 thawed motile spermatozoa:oocyte activated by 119 mM NaHCO(3) assured a fertilization rate of 74%.

Evaluation of fish sperm concentration using two counting chambers

A concentraçäo espermática do sêmen do peixe anostomídeo Leporinus elongatus (Valenciennes, 1849) foi estimada nas câmaras de Makler e de Neubauer, tendo-se obtido, respectivamente, 10,9 ± 2,8 ´ 10(9) e 11,9 ± 3,7 ´ 10(9) espermatozóides/ml de sêmen sem que ocorresse diferença significativa entre essas médias (P > 0,05). A estimativa da concentraçäo espermática do sêmen na câmara de Makler apresentou vantagens em relaçäo à da câmara de Neubauer. A contagem na câmara de Makler pôde ser feita sem a necessidade de se esperar pela deposiçäo dos espermatozóides no seu fundo. A baixa diluiçäo do sêmen (1:1.000) necessária para se estimar a concentraçäo espermática pode ser considerada também como vantagem pois altas diluiçöes (como na câmara de Neubauer) säo fontes consideráveis de erro. Dessa forma, a câmara de Makler pode ser utilizada na determinaçäo da concentraçäo espermática do sêmen de peixes e, possivelmente, de outros vertebrados, incluindo a de animais domésticos