The possibility of ice crystal structuring is attractive because it is possible to reach some structures of reduced dimensions capable of decrease the freezing damage to the minimal. The occurrences of needles or skeletons harmful to the frozen tissues demonstrate that the crystal face grown were unequal. The objective of the research is to the induce formation of smaller crystalline seeds, spherical or with a high symmetry level to increase the probability of original form. It was used: 1) anphiphilic substances, to affect the usual crystalline structure of the water; 2) substances with hydrogen bond possibility containing water in all directions in order to arrange symmetrical structures. Under the influence of sodium dodecyl sulfate (SDS), a surface-active agent, the ice crystals composed of spherical crystalline aggregates. Spherulitic crystals were obtained under the influence of valina amino acid, histidine, glicine and salt sulfate aluminum and ammonia. Polyhedric crystals with curve faces were obtained with the influence of the surface-active agent Triton ® X 100, Tween ® 80 and of the bile salt the quenodeoxicolic acid. The best result was obtained under the influence of phoroglucine. This of the triphenol was capable of producing small crystals with rounded extremities and in all stratum of the sample. In all the order solutions the spherical crystals appears in the superior stratum only. | Estruturar cristais de gelo é atrativo devido à possibilidade de diminuir os danos causados pelo congelamento ao mínimo. A ocorrência de agulhas ou esqueletos danosos às células significa que foram desiguais os crescimentos das faces do cristal. Objetivou-se com este trabalho induzir a formação de sementes cristalinas menores, arredondadas ou com alto grau de simetria, para aumentar a probabilidade de crescimentos geometricamente semelhantes à forma original e que elimine crescimentos desiguais nas faces do cristal de gelo. Foram usadas: 1) substâncias anfifílicas, para afetar a estruturação cristalina habitual da água e 2) substâncias com a possibilidade de formação de ligações de hidrogênio nas diversas direções e formar arranjos simétricos. Sob a influência do dodecil sulfato sódico (SDS), um tensoativo, os cristais de gelo cristalizaram na forma de agregados cristalinos esféricos. Cristais esferulíticos foram obtidos sob a influência dos aminoácidos valina, histidina, glicina e do sal sulfato de alumínio e amônio. Cristais poliédricos com faces curvas foram conseguidos sob a influência dos tensoativos Triton ® X 100, Tween ® 80 e de um sal biliar (ácido quenodeoxicólico). O melhor resultado foi obtido sob a influência da floroglucina. Esse trifenol produziu cristais pequenos, com extremidades arredondadas e em todas as camadas da amostra.

Estruturar cristais de gelo é atrativo devido à possibilidade de diminuir os danos causados pelo congelamento ao mínimo. A ocorrência de agulhas ou esqueletos danosos às células significa que foram desiguais os crescimentos das faces do cristal. Objetivou-se com este trabalho induzir a formação de sementes cristalinas menores, arredondadas ou com alto grau de simetria, para aumentar a probabilidade de crescimentos geometricamente semelhantes à forma original e que elimine crescimentos desiguais nas faces do cristal de gelo. Foram usadas: 1) substâncias anfifílicas, para afetar a estruturação cristalina habitual da água e 2) substâncias com a possibilidade de formação de ligações de hidrogênio nas diversas direções e formar arranjos simétricos. Sob a influência do dodecil sulfato sódico (SDS), um tensoativo, os cristais de gelo cristalizaram na forma de agregados cristalinos esféricos. Cristais esferulíticos foram obtidos sob a influência dos aminoácidos valina, histidina, glicina e do sal sulfato de alumínio e amônio. Cristais poliédricos com faces curvas foram conseguidos sob a influência dos tensoativos Triton ® X 100, Tween ® 80 e de um sal biliar (ácido quenodeoxicólico). O melhor resultado foi obtido sob a influência da floroglucina. Esse trifenol produziu cristais pequenos, com extremidades arredondadas e em todas as camadas da amostra.<br>The possibility of ice crystal structuring is attractive because it is possible to reach some structures of reduced dimensions capable of decrease the freezing damage to the minimal. The occurrences of needles or skeletons harmful to the frozen tissues demonstrate that the crystal face grown were unequal. The objective of the research is to the induce formation of smaller crystalline seeds, spherical or with a high symmetry level to increase the probability of original form. It was used: 1) anphiphilic substances, to affect the usual crystalline structure of the water; 2) substances with hydrogen bond possibility containing water in all directions in order to arrange symmetrical structures. Under the influence of sodium dodecyl sulfate (SDS), a surface-active agent, the ice crystals composed of spherical crystalline aggregates. Spherulitic crystals were obtained under the influence of valina amino acid, histidine, glicine and salt sulfate aluminum and ammonia. Polyhedric crystals with curve faces were obtained with the influence of the surface-active agent Triton ® X 100, Tween ® 80 and of the bile salt the quenodeoxicolic acid. The best result was obtained under the influence of phoroglucine. This of the triphenol was capable of producing small crystals with rounded extremities and in all stratum of the sample. In all the order solutions the spherical crystals appears in the superior stratum only.

Estruturar cristais de gelo é atrativo devido à possibilidade de diminuir os danos causados pelo congelamento ao mínimo. A ocorrência de agulhas ou esqueletos danosos às células significa que foram desiguais os crescimentos das faces do cristal. Objetivou-se com este trabalho induzir a formação de sementes cristalinas menores, arredondadas ou com alto grau de simetria, para aumentar a probabilidade de crescimentos geometricamente semelhantes à forma original e que elimine crescimentos desiguais nas faces do cristal de gelo. Foram usadas: 1) substâncias anfifílicas, para afetar a estruturação cristalina habitual da água e 2) substâncias com a possibilidade de formação de ligações de hidrogênio nas diversas direções e formar arranjos simétricos. Sob a influência do dodecil sulfato sódico (SDS), um tensoativo, os cristais de gelo cristalizaram na forma de agregados cristalinos esféricos. Cristais esferulíticos foram obtidos sob a influência dos aminoácidos valina, histidina, glicina e do sal sulfato de alumínio e amônio. Cristais poliédricos com faces curvas foram conseguidos sob a influência dos tensoativos Triton ® X 100, Tween ® 80 e de um sal biliar (ácido quenodeoxicólico). O melhor resultado foi obtido sob a influência da floroglucina. Esse trifenol produziu cristais pequenos, com extremidades arredondadas e em todas as camadas da amostra. | The possibility of ice crystal structuring is attractive because it is possible to reach some structures of reduced dimensions capable of decrease the freezing damage to the minimal. The occurrences of needles or skeletons harmful to the frozen tissues demonstrate that the crystal face grown were unequal. The objective of the research is to the induce formation of smaller crystalline seeds, spherical or with a high symmetry level to increase the probability of original form. It was used: 1) anphiphilic substances, to affect the usual crystalline structure of the water; 2) substances with hydrogen bond possibility containing water in all directions in order to arrange symmetrical structures. Under the influence of sodium dodecyl sulfate (SDS), a surface-active agent, the ice crystals composed of spherical crystalline aggregates. Spherulitic crystals were obtained under the influence of valina amino acid, histidine, glicine and salt sulfate aluminum and ammonia. Polyhedric crystals with curve faces were obtained with the influence of the surface-active agent Triton ® X 100, Tween ® 80 and of the bile salt the quenodeoxicolic acid. The best result was obtained under the influence of phoroglucine. This of the triphenol was capable of producing small crystals with rounded extremities and in all stratum of the sample. In all the order solutions the spherical crystals appears in the superior stratum only.

The aim of this study was to evaluate the ACP-102c extender in the cryopreservation of ram semen compared to tris-glucose-egg yolk (TRIS) extender and fresh semen. Forty-eight ejaculates were collected from four rams and two aliquots per ejaculate were taken for dilution and cryopreservation in ACP-102c or TRIS and a third aliquot used for the fresh semen analysis. Either the fresh semen and cryopreserved in both extenders were evaluated for viability, integrity of plasma and acrosomal membrane, hypoosmotic swelling test, DNA fragmentation and sperm motility. The extenders did not differ for sperm viability, acrosome and plasma membrane integrity, DNA fragmentation and quantitative and qualitative parameters of sperm velocity after thawing, but differed in hypoosmotic swelling test, total and progressive motility and lateral extent of the head as well as in all motility parameters evaluated (except linearity and straightness) after two hours of incubation at 37 oC. There was variability among rams in total and progressive motility of semen cryopreserved in ACP-102c after thawing. The ACP-102c extender showed less protection in the cryopreservation of ram sperm when compared to TRIS, but the improvement in its formulation and freezing protocols may make it an alternative to freezing ram semen.

ABSTRACTThe aim of this study was to determine the effects of various dilution ratios on thaw sperm motility, velocity and presence of spermatic abnormalities in the common carp. 20 males with three years of age, raised at the Departamento Nacional de Obras Contra as Secas (DNOCS) were used. From the collected semen a total of nine pools were formed. The semen from each pool was evaluated for motility, velocity and spermatic morphology before and after cryopreservation. The sperm cryopreservation was performed in medium ACP-104 + DMSO 10% diluted in 1:1 or 1:3 (sperm:extender). The samples were loaded into 0.25mL straws, frozen in liquid nitrogen vapor (Styrofoam) and stored in liquid nitrogen (-196 °C). There was no significant difference (P>0.05) between the ratios of dilution on the kinetic and morphological parameters of thawed semen from common carp. However, these parameters were significantly different (P<0.05) compared to fresh semen (control). Low velocity values were recorded in thawed semen (VCL 42.6-46.5μm/s; VSL 33.2-37.1μm/s; VAP 38.9-43.2μm/s), however, observed motility above 59% (59.8-67.8%), and low levels of spermatic abnormalities (19.3-19.5%) suggested that the ACP-104 can be used as a suitable medium for cryopreservation of common carp semen in any of the tested dilution ratios. | RESUMOO objetivo deste estudo foi avaliar o efeito de diferentes taxas de diluição seminal sobre a motilidade, velocidade e presença de anormalidades espermáticas no sêmen descongelado de carpa comum (Cyprinus carpio). Utilizaram-se 20 machos sexualmente maduros, pertencentes ao Departamento Nacional de Obras Contra as Secas (DNOCS). A partir do sêmen coletado, foi formado um total de nove pools. O sêmen de cada pool foi avaliado quanto à motilidade, velocidade e morfologia espermática antes e depois da criopreservação. A criopreservação seminal foi realizada em meio ACP-104 + DMSO 10% diluído em 1:1 ou 1:3 (sêmen:diluidor). As amostras foram envasadas em palhetas de 0,25mL, congeladas em vapor de nitrogênio líquido (caixa de poliestireno) e estocadas em nitrogênio líquido (-196 °C). Não houve diferença significativa (P>0,05) entre as taxas de diluições sobre os parâmetros cinéticos e morfológicos de sêmen descongelado de carpa comum. Entretanto, esses parâmetros apresentaram diferença significativa (P<0,05) em relação ao sêmen fresco (controle). Baixos valores de velocidade foram registrados no sêmen descongelado (VCL 42,6-46,5μm/s; VSL 33,2-37,1μm/s; VAP 38,9-43,2μm/s); contudo, observaram-se motilidades acima de 59% (59,8-67,8%) e baixos índices de anormalidades espermáticas (19,3-19,5%), sugerindo que o ACP pode ser usado como um meio favorável à criopreservação do sêmen de carpa comum em qualquer uma das taxas de diluição testadas.

Compararam-se as características cinéticas e morfológicas de espermatozoides caprinos congelados nos meios à base de ACP-101® e TRIS. Os diluentes utilizados foram: ACP-101® (+ 2,5% gema ovo + 7% glicerol) e TRIS (+ 20% gema ovo + 6,8% glicerol). Quarenta e oito ejaculados de quatro bodes foram coletados, avaliados, divididos em duas alíquotas e diluídos nos meios ACP-101® e TRIS, respectivamente, posteriormente congelados e, após 30 dias, descongelados. A avaliação da motilidade espermática por computador foi realizada aos 5, 60 e 120 minutos pós-descongelação. As características de motilidade espermática analisadas foram: motilidade total (MT) (%) e progressiva (MP) (%), velocidades média do trajeto do espermatozoide (VAP) (µm/s) e linear (VSL) (µm/s) e população de espermatozoides rápidos (ER) (%). As avaliações de morfologia espermática quantificaram a porcentagem de espermatozoides normais (N) e as alterações da cabeça (AC), da peça intermediária (API) e do flagelo (AF), aos cinco e 120 minutos pós-descongelação. O diluente TRIS apresentou resultados cinéticos mais elevados que o ACP-101® aos 60 e 120 minutos pós-descongelação. As AC aos 120 minutos pós-descongelação foram mais altas nos espermatozoides congelados em ACP-101®. Conclui-se que o diluente TRIS promoveu maior viabilidade in vitro dos espermatozoides caprinos pós-descongelação. | The aim of the work was to compare kinetic and morphologic characteristics of goat sperm frozen in diluent media based on ACP-101® and TRIS. The employed diluents were: ACP-101® (+ 2.5% egg yolk + 7% glycerol) and TRIS (+ 20% egg yolk + 6.8% glycerol). Forty eight ejaculates from four bucks were collected, assessed and divided into two aliquots and diluted into the experimental treatments ACP-101® and TRIS. The samples were frozen and after 30 days, thawed. The computer assisted spermatic motility evaluations were placed into 5, 60 and 120 minutes post-thawing. The motion parameters assessed were: total motility (MT) (%), progressive motility (MP) (%), average path velocity (VAP) (µm/s), straight linear velocity (VSL) (µm/s), and population of rapid spermatozoa (ER) (%). The morphologic parameters: normal spermatozoa (N), head alteration (AC), intermediary piece alteration (API), tail alteration (AF) were evaluated at 5 and 120 min post-thawing. The media based on TRIS showed kinetic results significantly superior to ACP-101® at 60 and 120min. After 120min post-thawing the AC was higher in frozen sperm in media based on ACP-101®. The TRIS media promoted better goat spermatozoa in vitro viability post-thawing.

It is known that temperature and thawing velocity can affect sperm quality during freeze-thawing procedures. Thus, the aim of this research was to compare different thawing protocols for canine semen extended in ACP-106® extender and cryopreserved. Ten ejaculates were collected from eight dogs and sperm rich fraction was evaluated regarding volume, concentration, motility, vigor, pH, morphology, acrosomal integrity and hyposmotic swelling test. Then, semen was extended in ACP-106® with 20% egg yolk and 6% glycerol, and frozen in 0.25 mL straws. After one week, one straw was thawed at 37°C/60s, another one at 55°C/5s and a third one at 75°C/8s, and the seminal characteristics were reevaluated. Similar results were observed for all the groups, except for sperm motility which was better preserved at 55°C/5s for up 30min than at 37°C/60s or 75°C/8s. From these results, it is concluded that canine semen extended in ACP -106® is better thawed at 55°C/5s.