Physiological routes from intra-uterine seminal contents to advancement of ovulation

Whole boar semen or seminal plasma has been demonstrated to advance the time of ovulation in gilts. As a means of clarifying this influence, the contribution of uterine lymphatics and their white cell populations has been examined. After duct visualisation with Evan's blue, lymph was sampled from a mesometrial vessel in eight pre-ovulatory gilts whose uterine lumen was infused simultaneously with whole semen in one ligated horn and saline in the contralateral ligated horn. Lymph was collected from cannulated vessels for periods of up to four hours under general anaesthesia. Thereafter, mesometrial lymph nodes, utero-tubal junction and uterine wall tissues were sampled. The proportion of nucleated cells in the sampled lymph increased towards the end of the collection period, but erythrocytes were found in all instances preventing a meaningful differentiation and identification of leukocytes. Prominent uterine lymph nodes were present in the mesometrium on both sides of the reproductive tract in 7 of 10 gilts. Differences in cellular contents were demonstrated between the side of the tract infused with semen and that infused with saline control. Two of 4 gilts had lower values for CD4 (Cluster Differentiation) and 3 of 6 gilts higher values for MHC II (Major Histocompatibility Complex) markers on the side challenged with semen. In contrast, values remained constant for CD8 but ranged widely for CD18. Immunohistochemical analysis of uterine tissue samples for MHC II+ cells revealed significant differences (P < 0.05) between the control and semen-treated ligated portions of the horns, as well as between the tissue sample of uterine wall and that from the utero-tubal junction, but there were no significant differences for CD4+ cells. It therefore remains plausible that semen-induced cytokines in the uterine lymph undergo counter-current transfer to the ipsilateral ovary and accelerate the final maturation of pre-ovulatory Graafian follicles

Fertilidade de sêmen suíno avaliada pelo teste de ligação dos espermatozóides a um substrato sintético

The objective of this work was to evaluate boar semen fertility by a sperm-binding assay to a synthetic substrate. Motility (MOT) and percentage of bound sperm (PSB) were evaluated after 5, 24, 48 and 72 hours of storage at 17°C. PSB was analyzed in solutions containing 6.25 or 12.5 million of spermatozoa/mL, with or without bovine serum albumin (BSA), processed from two to five ejaculates of four boars. Fifty eight gilts were inseminated, a single time, 24 hours after the beginning of estrus. There was a positive correlation (P = 0.0001; r = 0.33) between MOT and PSB. Higher percentages of PSB were observed with 12.5 million of spermatozoa/mL and in the presence of BSA (P < 0.05). After 72 hours, boar 3 showed lower PSB (P < 0.05) than the other boars. The cleavage (TC) and normal embryo rates did not differ among boars, but boar 3 showed less than 70.0% of TC belonging to the superior quartile while boars 1, 2 and 4 had more than 75.0%. After 24 hours of sperm storage, boars differ in their sperm binding to the synthetic substrate. Binding of swine spermatozoa to the synthetic substrate is higher in the presence of BSA and with the increase of spermatic concentration.O objetivo deste trabalho foi avaliar a fertilidade de sêmen suíno pelo teste de ligação de espermatozóides a um substrato sintético. A motilidade (MOT) e o porcentual de espermatozóides ligados (PEL) foram avaliados após 5, 24, 48 e 72 horas de armazenamento a 17ºC. O PEL foi determinado em soluções contendo 6,25 ou 12,5 milhões de espermatozóides/mL, com ou sem albumina sérica bovina (BSA), preparadas a partir de dois a cinco ejaculados de cada um dos quatro machos. Cinqüenta e oito leitoas foram inseminadas, uma vez, 24 horas após o início do estro. Houve correlação positiva (P = 0,0001; r = 0,33) entre a MOT e o PEL. O PEL foi maior com 12,5 milhões de  espermatozóides/mL e na presença de BSA (P < 0,05). Após 72 horas, o macho 3 apresentou PEL inferior ao dos outros três (P < 0,05). As taxas de clivagem (TC) e de embriões morfologicamente normais não diferiram entre indivíduos, mas o macho 3 apresentou menos de 70,0% de TC no quartil superior, enquanto os outros tiveram mais de 75,0%. Os machos diferem quanto à capacidade de ligação de seus espermatozóides ao substrato sintético, a partir de 24 horas de armazenamento do sêmen. A ligação dos espermatozóides ao substrato sintético é maior com a inclusão de BSA e com o aumento da concentração espermática

Pré-diluição e congelação de sêmen suíno em água de coco in natura, após três diferentes pré-tratamentos de incubação

Foram utilizados vinte e quatro ejaculados de cinco diferentes cachaços na congelação de sêmen suíno, sendo doze deles pré-diluídos em água de coco in natura e em Merck I, utilizando a lactose como diluidor de refrigeração e de congelação (experimento A) e doze pré-diluídos apenas com Merck I. Após três diferentes pré-tratamentos, a água de coco in natura foi utilizada como diluidor de refrigeração e de congelação, sendo a lactose utilizada como controle (experimento B). Seguiu-se a metodologia convencional de congelação de sêmen desta espécie -Tierärztliche Hochschule Hannover (grupo 1A-1B) e um novo processo com longo período de equilíbrio (grupos 2A-2B e 3A-3B). A qualidade do sêmen descongelado foi avaliada pela motilidade subjetiva (SMOT), motilidade computadorizada (CMOT) e morfologia das células com borda apical normal (NAR), após fixação em formol citrato. As porcentagens de espermatozóides com NAR foram 65% (grupo 1A), 71% (grupo 2A) e 75% (grupo 3A) para o sêmen pré-diluído em água de coco e 60% (grupo 1A), 68% (grupo 2A) e 68% (grupo 3A) para aquele pré-diluído com Merck I (experimento A); e 56% (grupo 1B), 68% (grupo 2B) e 73% (grupo 3B) para o sêmen congelado em água de coco e 60% (grupo 1B), 68% (grupo 2B e 3B) para o sêmen congelado em lactose (experimento B). Não havendo, portanto, diferença estatística entre os dois pré-diluentes e os dois diluentes de refrigeração e de congelação (p<0,05). Concluiu-se, portanto, que 1) os pré-tratamentos com longo período de equilíbrio têm melhor efeito na proteção do acrossoma do espermatozóide suíno e para manter a motilidade espermática e 2) a água de coco como pré-diluente e diluente para refrigeração e de congelação é semelhante ao Merck I (experimento A) e a lactose (experimento B), sendo portanto indicado para pré-diluir e congelar sêmen suíno

Fertilidade de sêmen suíno avaliada pelo teste de ligação dos espermatozóides a um substrato sintético

O objetivo deste trabalho foi avaliar a fertilidade de sêmen suíno pelo teste de ligação de espermatozóides a um substrato sintético. A motilidade (MOT) e o porcentual de espermatozóides ligados (PEL) foram avaliados após 5, 24, 48 e 72 horas de armazenamento a 17ºC. O PEL foi determinado em soluções contendo 6,25 ou 12,5 milhões de espermatozóides/mL, com ou sem albumina sérica bovina (BSA), preparadas a partir de dois a cinco ejaculados de cada um dos quatro machos. Cinqüenta e oito leitoas foram inseminadas, uma vez, 24 horas após o início do estro. Houve correlação positiva (P = 0,0001; r = 0,33) entre a MOT e o PEL. O PEL foi maior com 12,5 milhões de espermatozóides/mL e na presença de BSA (P<0,05). Após 72 horas, o macho 3 apresentou PEL inferior ao dos outros três (P<0,05). As taxas de clivagem (TC) e de embriões morfologicamente normais não diferiram entre indivíduos, mas o macho 3 apresentou menos de 70,0% de TC no quartil superior, enquanto os outros tiveram mais de 75,0%. Os machos diferem quanto à capacidade de ligação de seus espermatozóides ao substrato sintético, a partir de 24 horas de armazenamento do sêmen. A ligação dos espermatozóides ao substrato sintético é maior com a inclusão de BSA e com o aumento da concentração espermática.The objective of this work was to evaluate boar semen fertility by a sperm-binding assay to a synthetic substrate. Motility (MOT) and percentage of bound sperm (PSB) were evaluated after 5, 24, 48 and 72 hours of storage at 17°C. PSB was analyzed in solutions containing 6.25 or 12.5 million of spermatozoa/mL, with or without bovine serum albumin (BSA), processed from two to five ejaculates of four boars. Fifty eight gilts were inseminated, a single time, 24 hours after the beginning of estrus. There was a positive correlation (P = 0.0001; r = 0.33) between MOT and PSB. Higher percentages of PSB were observed with 12.5 million of spermatozoa/mL and in the presence of BSA (P<0.05). After 72 hours, boar 3 showed lower PSB (P<0.05) than the other boars. The cleavage (TC) and normal embryo rates did not differ among boars, but boar 3 showed less than 70.0% of TC belonging to the superior quartile while boars 1, 2 and 4 had more than 75.0%. After 24 hours of sperm storage, boars differ in their sperm binding to the synthetic substrate. Binding of swine spermatozoa to the synthetic substrate is higher in the presence of BSA and with the increase of spermatic concentratio

Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

The role of antibiotics (AB) in semen extenders as a potential contribution to the global antimicrobial resistance threat is emerging. Here, we establish an AB-free hypothermic preservation strategy for boar semen and investigate its impact on sperm function, microbial load and fertility after artificial insemination (AI). Spermatozoa (12 boars) preserved in AB-free AndroStar Premium extender at 5 °C maintained high motility, membrane integrity, and a low DNA-fragmentation index throughout 72 h storage and results did not significantly differ from controls stored at 17 °C in extender containing AB (p = 0.072). Likewise, kinetic response of spermatoza to the capacitation stimulus bicarbonate during 180 min incubation in Tyrode's medium did not differ from 17 °C-controls. In a competitive sperm oviduct binding assay, binding indices did not differ between semen stored for 72 h AB-free at 5 °C and 17 °C-controls (n = 6 boars). Bacterial load  0.05). In conclusion, a hypothermic semen preservation strategy is presented which offers antibiotic-free storage of boar semen doses

Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

The role of antibiotics (AB) in semen extenders as a potential contribution to the global antimicrobial resistance threat is emerging. Here, we establish an AB-free hypothermic preservation strategy for boar semen and investigate its impact on sperm function, microbial load and fertility after artificial insemination (AI). Spermatozoa (12 boars) preserved in AB-free AndroStar Premium extender at 5 °C maintained high motility, membrane integrity, and a low DNA-fragmentation index throughout 72 h storage and results did not significantly differ from controls stored at 17 °C in extender containing AB (p = 0.072). Likewise, kinetic response of spermatoza to the capacitation stimulus bicarbonate during 180 min incubation in Tyrode's medium did not differ from 17 °C-controls. In a competitive sperm oviduct binding assay, binding indices did not differ between semen stored for 72 h AB-free at 5 °C and 17 °C-controls (n = 6 boars). Bacterial load  0.05). In conclusion, a hypothermic semen preservation strategy is presented which offers antibiotic-free storage of boar semen doses