Conservação de peças anatômicas em cloreto de sódio: um estudo com produto de fecundação / Conservation of anatomical parts in sodium chloride: a study with fertilization product / Conservación de partes anatómicas en cloruro sódico: un estudio con producto de fertilización

Introdução: Os materiais de origem humana geralmente são conservados em formaldeído, para possibilitar o estudo da anatomia humana, tal conservante possui baixo custo e boa fixação, contudo é toxico. Diante do exposto é necessário, o estudo de outros métodos de conservação, menos prejudiciais, como a solução de NaCl 30%. Objetivo: Comparar a conservação de peças anatômicas em solução de NaCl à 30% e formaldeído a 10%. Método: Pesquisa experimental, exploratória e descritiva, realizada com dois produtos de abortamento, no laboratório de anatomia de uma universidade pública, no estado do Paraná/BR. Foi realizada fixação em solução de formol 10%, em seguida uma amostra foi lavado em água corrente e armazenado em solução de NaCl à 30%. Após 6 meses da conservação em solução salina, foram coletadas amostras, estas foram submetidas a análise de crescimento bacteriano. Avaliou-se tonalidade e turgor cutâneo, odor e peso, bem como crescimento bacteriano. O estudo seguiu os preceitos éticos (CAAE: 53740121.9.0000.9247). Resultados: Foram realizadas observações após 24h, 7, 30, 60, 90 e 180 dias. O feto em solução de NaCl não possui odor, e diminuição do turgor da pele. Ambas a amostras não apresentaram crescimento bacteriano. Considerações finais: A solução de NaCl a 30% desidrata a pele, mas não altera significativamente a forma e estrutura, ainda não possui odor e nem toxicidade, o que garante benefícios a saúde de quem os manipula, bem como tal concentração de NaCl inibe de forma efetiva o crescimento bacteriano nos tecidos e na própria solução, se demostrando eficaz na conservação.

Introduction: The materials of human origin are usually preserved in formaldehyde, to enable the study of human anatomy, this preservative has low cost and good fixation, however it is toxic. Therefore, it is necessary to study other less harmful preservation methods, such as 30% NaCl solution. Objective: To compare the preservation of anatomical specimens in 30% NaCl solution and 10% formaldehyde solution. Method: Experimental, exploratory and descriptive research, carried out with two abortion products, in the anatomy laboratory of a public university, in the state of Paraná/BR. Fixation in 10% formaldehyde solution was performed, after which a sample was washed in running water and stored in a 30% NaCl solution. After 6 months of preservation in saline solution, samples were collected and submitted to bacterial growth analysis. Skin tone and turgor, odor, weight, and bacterial growth were evaluated. The study followed the ethical precepts (CAAE: 53740121.9.0000.9247). Results: Observations were made after 24h, 7, 30, 60, 90 and 180 days. The fetus in NaCl solution had no odor, and decreased skin turgor. Both samples showed no bacterial growth. Final considerations: The 30% NaCl solution dehydrates the skin, but does not alter significantly the shape and structure, and also has no odor or toxicity, which guarantees health benefits to those who handle them, and such concentration of NaCl inhibits effectively the bacterial growth in the tissues and in the solution itself, proving to be effective in conservation.

Introducción: Los materiales de origen humano suelen conservarse en formol, para posibilitar el estudio de la anatomía humana, este conservante tiene bajo coste y buena fijación, sin embargo es tóxico. Por ello, es necesario estudiar otros métodos de conservación menos nocivos, como la solución de NaCl al 30%. Objetivo: Comparar la conservación de especímenes anatómicos en solución de NaCl al 30% y en solución de formaldehído al 10%. Método: Investigación experimental, exploratoria y descriptiva, realizada con dos abortos, en el laboratorio de anatomía de una universidad pública, en el estado de Paraná/BR. Fue realizada fijación en solución de formaldehído al 10%, después de lo cual la muestra fue lavada en agua corriente y almacenada en solución de NaCl al 30%. Tras 6 meses de conservación en solución salina, se recogieron las muestras y se sometieron a análisis de crecimiento bacteriano. Se evaluaron el tono y la turgencia de la piel, el olor, el peso y el crecimiento bacteriano. El estudio siguió los preceptos éticos (CAAE: 53740121.9.0000.9247). Resultados: Las observaciones se realizaron después de 24h, 7, 30, 60, 90 y 180 días. El feto en solución de NaCl no tenía olor, y la turgencia de la piel disminuyó. Ambas muestras no mostraron crecimiento bacteriano. Consideraciones finales: La solución de NaCl al 30% deshidrata la piel, pero no altera significativamente la forma y estructura, además no tiene olor ni toxicidad, lo que garantiza beneficios para la salud de quienes los manipulan, y dicha concentración de NaCl inhibe eficazmente el crecimiento bacteriano en los tejidos y en la propia solución, demostrando ser eficaz en la conservación.

Influence of category-heifers, primiparous and multiparous lactating cows-in a large-scale resynchronization fixed-time artificial insemination program

This study was conducted to evaluate the influence of category (heifers, primiparous or multiparous cows) on pregnancy rates in a large scale resynchronization ovulation program. Nelore heifers (n = 903), primiparous lactating cows (n = 338) and multiparous lactating cows (n = 1,223) were synchronized using a conventional protocol of estradiol/P4-based fixed-time artificial insemination (FTAI). Thirty days after ultrasonography, females who failed the first FTAI were resynchronized with the same hormonal protocol prior to a second FTAI. The pregnancy status of each cohort was evaluated by ultrasonography 30 days after each FTAI. The average conception rate after the first FTAI and resynchronization was 80.5%. Heifers had a higher conception rate (85%) than primiparous (76%) or multiparous cows (78%; p = 0.0001). The conception rate after the first FTAI was similar among heifers (57%), primiparous cows (51%) and multiparous cows (56%; p = 0.193). After the second FTAI, heifers exhibited a higher conception rate (66%) than primiparous or multiparous cows (51%; p = 0.0001). These results demonstrate the feasibility of resynchronization in large beef herds for providing consistent pregnancy rates in a short period of time. We also demonstrated that ovulation resynchronization 30 days after FTAI is particularly effective for heifers, providing a conception rate of up to 66%.

Influence of L-arginine during bovine in vitro fertilization.

The objective of this work was to evaluate the effect of using L-arginine during in vitro fertilization (IVF) on in vitro embryonic development using Bos taurus and Bos indicus semen. Effect of different concentrations (0, 1, 10 and 50 mM) of L-arginine, added to the IVF medium, was evaluated on the fertilization rate at 18 h post-fertilization (hpf), NO3-/NO2- production during IVF by the Griess colorimetric method (30 hpf), cleavage and blastocyst rates (on Day 2 and Day 7 of culture, respectively) and total blastocyst cell number (Day 7 of culture). The results reveal that the addition of 50 mM L-arginine to IVF medium, with either Bos taurus or Bos indicus spermatozoa, decreased the cleavage rate and blastocyst rate compared to the control group. Other concentrations did not affect embryo production. However, 1 mM L-arginine with Bos indicus semen increased the proportion of hatched blastocysts. These results indicate that high L-arginine concentrations may exhibit toxic effects on bovine gametes during in vitro fertilization.

Effects of progestagen exposure duration on estrus synchronization and conception rates of crossbreed ewes undergoing fixed time artificial insemination

Synchronization of estrus and ovulation are of paramount importance in modern livestock improvement programs. These methods are critical for assisted reproduction technologies, including artificial insemination and embryo transfer, that can increase productivity. In the current study, subcutaneous implants containing norgestomet were placed for long (14 days), medium (9 days), and short (5 days) periods of time in 70 crossbred ewes undergoing fixed-time artificial insemination. The resulting effects on estrus synchronization and conception rates were subsequently evaluated. Among the synchronized ewes, 85.7% (60/70) underwent estrus over a period of 72 h after progestagen treatment ceased. The shortest mean interval between withdrawal of the device and onset of estrus (34.2 +/- 8.9 h) was observed in the G14 days of P4 group (p < 0.05). The conception rate of the G14 days of P4 group was statistically higher than that of the other groups (83.3% vs. 60.9% vs. 47.8%; p < 0.05). In conclusion, 14 days of norgestomet treatment produced higher conception rates and a greater number of pregnancies at the beginning of the breeding season.

effect of retinoic acid on in vitro maturation and fertilization rate of mouse germinal vesicle stage oocytes

Retinoids are recognized as important regulators of cell differention and tissue function, Previous studies, performed both in vivo and in vitro, indicate that retin-oids influence several reproductive events. In this study, we investigated the effect of all-trans retinoic acid [t-RA] on maturation and fertilization rate of immature oocytes [germinal vesicle]. Germinal vesicle [GV] oocytes were recovered from 4-6 week old female mice 48 hours after injection of 10 IU pregnant mare serum gonadotropin [PMSG]. Collected oocytes were divided into seven groups: control, sham and five experimental groups. t-RA at concentrations of 1, 2, 4, 6, 8 jjM were added to oocyte maturation medium in the experimental groups. The maturation rate was recorded after 24 hours of culture in a humidified atmosphere of 5% CO[2] at 37°C. Fertilization and developmental rates of matured oocytes were recorded after in vitro fertilization [IVF] and 24 hour culture. The rate of oocytes that developed to the metaphase II stage of maturation significantly increased with 2 and 4 microM t-RA compared to the control and sham groups [p<0.05]. In addition, the number of fertilized oocytes was significantly higher in 4 microM retinoic acid compared to the control [p<0.05], but the difference between the number of fertilized oocytes which developed to the 2-cell stage was not significant between the two groups. The results show that t-RA enhanced mouse oocyte maturation in vitro and improved fertilization and development rates in a dose dependent manner

[Effect of PGF[2] alpha injection on conception rate of insemenated repeat breeder cows during summer warm condition]

It has been demonstrated that heat stress has a detrimental effect on conception rate of dairy cows, via early embryonic death. To reduce heat stress, providing shade, fan, appropriate diet and application of ovulation induction agents [GnRH and hCG] is recommended. Endogen PGF[2]alpha has positive effect on ovulation. Repeat Breader. The aim of the present study was to evaluate the role of PGF[2] alpha injection on reproductive performance of Repeat Breader [RB] dairy cows in heat stress condition at artificial insemination [AI]. This study was conducted in two large dairy farms in Tehran province during the summer 1386-87. All cows were inseminated [A M -PM] based on heat detection at 12 hours after observed standing heat. Mean temperature and humidity were recorded throughout the study. In this study 150 RB cows were randomly divided into three groups. Group A [n=50] received PGF[2] alpha simultaneously with AI, group B [n=50] received GnRH at the time of AI and group C [n=50] did not receive any treatment and served as control. Based on rectal palpation on 40-50 days of AI, 5 cows of group A, 9 of group B and 10 group C were pregnant. There were not any significant differences among groups [p>0.05]. The results revealed that simultaneous using of PGF[2]alpha or GnRH with AI has no beneficial effects on conception rate in RB cows

[Effects of MDMA [ecstasy] on oocyte quality and fertilization rate in mice]

Ecstasy drug abuse is relatively common among the youth undergoing psychosocial stresses. Categorized as a stimulant drug, ecstasy has nervous, cardiac and immunological side-effects. The endocrine system is not spared from the harmful effects of the drug and it could influence ovulation by exerting its effects on the hypothalamic-hypophyseal-gonadal axis. The aim of this study was to evaluate the effects of ecstasy on the quality of oocytes and on fertilization rate in mice. Thirty female NMRI mice were divided into three groups, after checking for their estrous cycles. At the beginning of each cycle, ecstasy was administered intraperitoneally to the animals in groups A [a minimum dose of 5 mg/kg/day] and B [a maximum dose of 20 mg/kg/day]. At the end of the second day, 10 IU PMSG, and at the end of the 4th day, 10 IU HCG were administered intra-peritoneally. The mice in group C underwent the same procedures except receiving ecstasy. All the mice were sacrificed on the fifth day to aspirate their oocytes for further evaluation. In-vitro fertilization was done using mature or MII oocytes and fertilization was checked after 5 hours. The results were later statistically analyzed. The number of retrieved oocytes was similar between the experimental and control groups. However, the rate of mature oocytes [MII] was significantly higher [p=0.0001] in the controls [31.2%] than the mice in groups A [15.2%] and B [12.8%]. Moreover, fertilization rate was significantly higher in the controls than the experi-mental groups. Administration of Ecstasy in mice can affect both oocyte maturity and the fertilization potential of oocytes. The overall effects of ecstasy on embryo development and implantation needs further studies to reach a definitive conclusion

Effects of azadirachtin on mortality, fertilization, and swimming speed in larvae of the cane toad, Bufo marinus (Anura: Bufonidae).

Experiments were conducted to assess the effects of azadirachtin (AZ) on survival capacity, fertilization success, and swimming speed in larval stages of the cane toad, Bufo marinus . LC50 values (96-hr) for various larval stages exposed to 0.50 mg/l AZ were as follows: 5.35 (stage 12), 5.77 (stage 16), 787 (stage 20), 13.43 (stage 24), and 21.54 (stage 30). Tadpoles did not exhibit any apparent disorientation or production of mucous in response to AZ concentrations ranging from 0.3 to 0.5 mg/l. Mortality was significantly higher at 0.50 mg/l AZ than at lower concentrations. At 0.50 mg/l, mortality rate decreased significantly from 36% for stage 12 tadpoles, to 24% for stage 20 tadpoles. Mortality for early stages (stage 12) dropped significantly, from 36% at 0.50 mg/l, to 21% at 0.40 mg/l AZ There were no significant differences in percent mortality for any developmental stage at an AZ concentration of 0.3 mg/l, or for controls (no AZ). Exposure of gametes to 0.30 mg/l AZ did not reduce fertilization success as compared to controls (89.8 vs. 83.7%). At 0.40 mg/l, percent fertilization decreased significantly (68.8%). This effect was even more pronounced at 0.50 mg/l. No significant difference was found in swimming speeds between controls and subjects exposed to 0.30 mg/l AZ. However, at a concentration of 0.40 mg/l, tadpoles required a significantly longer period of time to swim a distance of 140 cm as compared to controls. The potential impact of AZ on amphibian populations is discussed.

Predictors of successful conception in a clinical trial of 30 patients with primary amenorrheic hypogonadotrophic hypogonadism treated by ovulation induction

A clinical trial was designed to assess the association between the treatment of infertile females with Hypogonadotrophic Hypogonadism and their fertility state considering conception as the desired outcome. Serum hormones, LH, FSH and estradiol, endometrial thickness and count of active ovarian follicles were assessed by U/S to evaluat their role in anticipating a successful conception. Standard protocols of ovulation induction were used in a clinical trial [historical cohort design] of 30 patients with primary hypogonadotrophic hypogonadism. For ethical reasons no attempt was made to fix the number of induction courses given to the female patient [such a variability is suspected to confound the result to a limited extent], so a female was followed up with several courses of ovulation induction [according to the advice of the physician in charge] for a median duration of I year with conception as the final outcome of interest. Four serum hormonal markers [FSH, LH, Prolaction and estradiol] were assessed before and 12-14 days after initiating therapy. Transvaginal U/S was done to measure endometrial thickness and the number of dominant follicles in both ovaries after 12-14 days of treatment. Serum estradiol concentration, endometrial thickness and number of dominant follicles were significant predictors of successful conception. Using endometrial thickness as a decision rule to predict successful conception at a cutoff value of 7 mm, considering the rule as positive if equal or greater than 7 mm, resulted in a positive predictive value [PPV] of 50% [Relative risk of having conception was 4.3 times]. The confidence in a positive rule was increased to 71.4% [PPV =11.4%] when the cutoff value was set higher at 8 mm endometrial thickness. Using the number of dominant follicles as a decision rule at a cutoff value of 4 resulted in a PPV of 60%. This PPV was further increased to 72.7%] when the cutoff value was set higher at 5. An U/S finding of 5 or more dominant follicles increased the.probability of having conception by 12.6 times. A serum estradiol concentration at day 12 of 114 pgm/ml and higher predicted successful conception with a PPV of 50% [relative risk of having conception was 7], increasing this cutoff value of serum estradiol to a maximum of 498.5 pgm/ml increased the PPV to 66.7%. a clinician can predict successful conception after the first 2 weeks of initiating therapy with a moderate degree of confidence depending on serum estradiol, endometrial thickness and number of dominant follicles in the ovary. Other factors however need to be considered to increase the confidence of prediction, such as the type of insemination technique, sperm count of husband. A larger sample size and a survival analysis design is needed to assess the exact risk of having conception after each course of treatment

Effect of eCG and GnRH on induction of ovarian activity and conception rate in post-partum ewes

Eighteen lactating Baladi ewes were divided into four groups. Group 1 [five ewes] received progesterone releasing intravaginal sponge containing 60 mg medroxy progesterone acetate [MAP]. Group 2 [five ewes] received a home made progesterone releasing intravaginal sponge containing 500 mg progesterone. Group 3 [four ewes] as in group 1 plus 750 IU equine chorionic gonadotropin [eCG] at sponge withdrawal. Group 4 [four ewes] was treated as in group 3 plus 100 mug GnRH analogue during estrus. In all groups, sponge remained in place for 12 days. Blood samples were collected before treatment, during estrus and 7, 14, 21 and 28 days post estrus. Progesterone levels were determined using RIA. The analysis of data showed that 60, 0, 100 and 100% of ewes in the 4 groups responded to the treatment after 72.00 +/- 0.00, 54.00 +/- 12.00 and 40.50 +/- 9.00 hr for groups 1, 3 and 4, respectively. Duration of estrus averaged 21.60 +/- 5.37, 16.50 +/- 5.74 and 15.00 +/- 6.00 hours for groups 1, 3 and 4, respectively. One ewe had conceived in groups 1, 3 and 4. Progesterone levels were higher in the responding than in non responding animals, and in pregnant than in non-pregnant ewes. In conclusion, treatment with 500 mg natural progesterone failed to induce estrus in postpartum ewes, while, eCG increase the incidence of estrus and neither GnRH nor eCG could improve ewe fertility during postpartum period