Reproductive toxicology and clastogenic evaluation in mice of a phytotherapeutic formulation obtained from Cinchona calisaya Weddel (Rubiaceae) used in Brazilian folk medicine as female fertility stimulant

Ethnopharmacological relevance: In Brazil, a phytotherapeutic preparation produced from a standardized tincture of Cinchona calisaya Weddel such that each mL of product contains 400 mu g of quinine, known in Portuguese as Agua Inglesa (R) (English water), is indicated by the manufacturer as a tonic, appetite stimulant, and digestive. However, this preparation has long been used in folk medicine as a female fertility stimulant. Despite its widespread use in folk medicine to stimulate female fertility, no study has been undertaken to assess the potential teratogenic and genotoxic effects of this phytotherapeutic preparation. The aim of the present study was to investigate possible toxic reproductive effects in mice caused by exposure to Agua Inglesa (R), either before mating or during the pre- and post-embryo implantation periods. The genotoxic potential was evaluated using the micronucleus assay.Material, Methods, and Results: Virgin female mice, with at least one estrous cycle evidenced by vaginal cytology, were divided into five groups of 15 individuals each (Group I - control, Group II - treated with ethanol solution at 16%, Groups III, IV and V treated with phytotherapeutic preparation at 1.5 mL/kg/day, 3.0 mL/kg/day and 4.5 mL/kg/day, respectively). After the first 28 days of treatment, females were caged individually with adult fertile males. Pregnant females continued to receive treatment for seven days (preimplantation period). Body weight was recorded weekly during treatment. Signs of toxicity (weight loss, food intake, piloerection, apathy, prostration, diarrhea, seizures, behavioral changes, and locomotion) were also observed. The females were sacrificed on the 15th day of pregnancy, uterine horns were evaluated for implantation, and the placental index was recorded. In the micronucleus test, 2000 polychromatic erythrocytes (PCE) per animal, obtained from bone marrow, were scored. Results The results showed that exposure of the females during the pre- and post-implantation periods did not significantly alter the reproductive capacity (p < 0.05); however, in higher dose (three times human dose)reduction of fetal weight was observed. There was no difference between the control and phytotherapeutic preparation (p > 0.05) in terms of the average number of micronucleated polychromatic erythrocytes.Conclusions: Although folk medicine suggests that the Agua Inglesa (R) preparation is useful as a female fertility stimulant, no such effect was confirmed in mice. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Effects of atrazine on female Wistar rats: Morphological alterations in ovarian follicles and immunocytochemical labeling of 90 kDa heat shock protein

Fertility in female mammals may be affected by a variety of endocrine disrupters present in the environment. Herbicide atrazine is an example of endocrine disrupter employed in agriculture, which disrupts estrous cyclicity in rats. Aiming to characterize morphologically the effect of low and sublethal doses of atrazine on the ovaries of Wistar rats, in an effort to determine the possible intrafollicular target site through which this herbicide acts adult females were submitted to both subacute and subchronic treatments. Additionally, immunocytochemical labeling of 90 kDa heat shock protein (HSP90) was performed in order to evaluate the role played by this protein in the ovary, under stressed conditions induced by herbicide exposure. The results indicated that atrazine induced impaired folliculogenesis, increased follicular atresia and HSP90 depletion in female rats submitted to subacute treatment, while the subchronic treatment with low dose of atrazine could compromise the reproductive capacity reflected by the presence of multioocytic follicle and stress-inducible HSP90. © 2007 Elsevier Ltd. All rights reserved.

Amifostine reduces the seminiferous epithelium damage in doxorubicin-treated prepubertal rats without improving the fertility status

Background: Amifostine is an efficient cytoprotector against toxicity caused by some chemotherapeutic drugs. Doxorubicin, a potent anticancer anthracycline, is known to produce spermatogenic damage even in low doses. Although some studies have suggested that amifostine does not confer protection to doxorubicin-induced testicular damage, schedules and age of treatment have different approach depending on the protocol. Thus, we proposed to investigate the potential cytoprotective action of amifostine against the damage provoked by doxorubicin to prepubertal rat testes (30-day-old) by assessing some macro and microscopic morphometric parameters 15, 30 and 60 days after the treatment; for fertility evaluation, quantitative analyses of sperm parameters and reproductive competence in the adult phase were also carried out.Methods: Thirty-day-old male rats were distributed into four groups: Doxorubicin (5 mg/kg), Amifostine (400 mg/kg), Amifostine/Doxorubicin (amifostine 15 minutes before doxorubicin) and Sham Control (0.9% saline solution). Standard One Way Anova parametric and Anova on Ranks non-parametric tests were applied according to the behavior of the obtained data; significant differences were considered when p < 0.05.Results: The rats killed 30 and 60 days after doxorubicin treatment showed diminution of seminiferous epithelium height and reduction on the frequency of tubular sections containing at least one type of differentiated spermatogonia; reduction of sperm concentration and motility and an increase of sperm anomalous forms where observed in doxorubicin-treated animals. All these parameters were improved in the Amifostine/Doxorubicin group only when compared to Doxorubicin group. Such reduction, however, still remained below the values obtained from the Sham Control group. Nevertheless, the reproductive competence of doxorubicin-treated rats was not improved by amifostine pre-administration.Conclusions: These results suggest that amifostine promotes a significant reduction of the doxorubicin long-term side effects on the seminiferous epithelium of prepubertal rats, which is reflected in the epidydimal fluid parameters in the adult phase. However, fertility status results suggest that such protection may not be effective against sperm DNA content damage. Further investigation of sperm DNA integrity must be carried out using amifostine and doxorubicin-treated experimental models. © 2010 Vendramini et al; licensee BioMed Central Ltd.

Aspectos biológicos de Cycloneda sanguínea (Coleóptera: Coccinellidae) alimentadas con pulgones criados en algodón transgénico Bollgard I

Cycloneda sanguinea is an important biological control agent of aphid cotton. The objective of this study was to evaluate the biological aspects of C. sanguinea fed with nymphs and adults of Aphis gossypii reared in DeltaOPAL and NuOPAL cultivars (Bollgard I). The experiment was conducted in the laboratory under controlled temperature (25 +/- 1 degrees C), humidity (70 +/- 10%) and photophase (12 hours). The experimental design was completely randomized, and treatments aphids fed on the two cultivars, and replicates 40 individuals. Each larva was individually until adult emergence of the predator in Petri dishes of 9 cm in diameter with moistened filter paper. For evaluation of individual adults were five couples newly emerged in plastic pots 12 cm in diameter and 9 cm tall, with filter paper for oviposition. The parameters evaluated were: duration and viability of the larval stage, pupal stage duration and survival, adult longevity, reproductive period, post-reproductive and total number of eggs per female. The duration of the larval stage of C. sanguinea was lower when fed aphids reared on the cultivar NuOPAL, while adult longevity and the reproductive period were higher. The female fertility and egg viability were also lower.

Efeito de sinigrina aplicada em folhas de brássicas sobre características biológicos de Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Correlações genéticas entre medidas de perímetro escrotal e características produtivas e reprodutivas de fêmeas da raça Nelore

Com objetivo de estimar parâmetros genéticos para perímetro escrotal medido aos 12 (PE12) e aos 18 meses de idade (PE18) de machos e peso a um ano de idade (P365), peso ao primeiro parto (PPP), idade ao primeiro parto (IPP), dias para o primeiro parto (DP) e duração da primeira gestação (DG) medidos em fêmeas, além de correlações entre perímetro escrotal e as demais características, foram analisadas medidas de 9.932 fêmeas e medidas de perímetro escrotal de 14.521 machos Nelore provenientes de 28 rebanhos das regiões Sul e Sudeste do País. Os componentes de (co)variância foram estimados utilizando-se o método da máxima verossimilhança restrita, sob modelo animal, em análises bi-características. As herdabilidades estimadas foram altas para PE12 (0,25 a 0,26), PE18 (0,35 a 0,37), P365 (0,35 e 0,38), PPP (0,40 e 0,41) e DG (0,19 e 0,20) e de menor magnitude para IPP (0,14 e 0,15) e DP (0,05 e 0,06). As correlações genéticas estimadas entre PE12 e P365, PPP, IPP, DP e DG foram de 0,28; 0,30; -0,15; -0,04 e 0,10, respectivamente. As correlações genéticas entre PE18 e P365, PPP, IPP, DP e DG, no entanto, foram de 0,29; 0,32; -0,21; -0,08 e 0,14, respectivamente. Esses resultados sugerem melhorias na eficiência reprodutiva de fêmeas, principalmente quando os rebanhos são selecionados para PE18.

The fibroblast growth factor family: involvement in the regulation of folliculogenesis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Reproductive effects in male rats exposed to diuron

Diuron is a ureic herbicide considered to have very low toxicity. The present study evaluated several aspects of reproductive toxicity of diuron in adult male rats. Diuron was diluted in corn oil and administered by oral gavage to groups of 18-20 rats at doses of 0, 125 or 250 mg/kg per day for 30 days; the control group received only the corn oil vehicle. At the end of the treatment period, approximately half the animals from each group were assigned to one of two terminal assessment lines: (1) reproductive organ, liver and kidney weights; measurement of diuron concentrations in liver and kidney; plasma testosterone determinations; evaluation of daily sperm production per testis; sperm number and sperm transit time in the epididymis; or (2) sexual behavior assessment during cohabitation with a receptive female; fertility and pregnancy outcome after natural mating; testicular, epididymal, kidney and liver histopathology; sperm morphology. After 30 days of oral diuron treatment, there were no treatment-related changes in body weights, but dose-related diuron residues were detected in the liver of all treated rats and absolute and relative liver weights were increased in both groups. There were no statistically significant differences between the treated and control groups obtained in plasma testosterone concentrations, or in parameters of daily sperm production, sperm reserves in the epididymis, sperm morphology or measured components of male sexual behavior. on the other hand, the number of fetuses in the litters from diuron-treated rats was slightly smaller than litters from control rats. Therefore, although the results did not indicate that diuron exposure resulted in direct male reproductive toxicity in the rat, they suggest that additional studies should be undertaken to investigate the possible effects on fertility and reproductive performance. (c) 2006 Elsevier B.V. All rights reserved.

Considerações sobre a reprodução de duas espécies de raias (Myliobatiformes, Potamotrygonidae) na região do Alto Rio Paraná, Sudeste do Brasil

This article presents preliminary information about the reproduction of two species of freshwater stingrays of the genus Potamotrygon, found in the Upper Parana River (Southwest Brazil), where these animals are exotic. Males of P. motoro and P. falkneri become sexually mature around 270 mm and 260 mm of disc width (DW), respectively. Females become sexually mature around 330 mm of DW in P. motoro and 325 mm in P. falkneri. In both species, females are bigger and heavier than males, reaching about 700 mm of DW and 20 kg. Copulating has not been recorded, but one courtship ritual has been observed during the dry season. Mature males present a different dentition from females and young males, which is probably related to their reproductive behavior instead of to their feeding habit. The female fertility has varied from one to three offspring. A miscarriage has been observed at different stages of embryonic development always when pregnant females were captured. In spite of the great history of anthropization of the study area, it seems that hydrological cycles are related to the reproduction of stingrays. Nevertheless, it would be necessary to conduct deeper studies to verify or not this influence.

Classificação de cultivares de brássicas com relação à resistência à traça-das-crucíferas e à presença de glucosinolatos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise genética da habilidade de permanência em fêmeas da raça Nelore

O objetivo deste estudo foi verificar a possibilidade da característica habilidade de permanência (HP) de matrizes ser utilizada como critério de seleção na raça Nelore. A HP foi definida como a probabilidade de uma vaca parir, no rebanho, na idade de seis anos ou depois desta idade, dado que ela teve uma parição em data anterior. Foram analisadas informações de 55.682 animais. Utilizou-se a amostragem de Gibbs para estimar os componentes de variância e um modelo de limiar de máximo a posteriori para predizer os valores genéticos. A análise forneceu estimativa posterior de herdabilidade e desvio-padrão de 0,21 ± 0,00 e tendência genética, média por ano, de 0,14% para HP. A facilidade de mensuração da característica, a estimativa de herdabilidade e a tendência indicam que a utilização desta característica como critério de seleção pode contribuir para o aumento da fertilidade do rebanho.

Influence of salivary glands extirpation on procreation in rats

The influence of extirpation of the submandibular and sublingual, parotid and all major salivary glands on pregnancy of rats was studied. The statistical analysis of the results indicated that there are statistically significant differences in the number of offsprings before and after extirpation of the salivary glands. The parotidectomized female rats presented the smallest number of offsprings as compared with other groups and presented some progressive sterility after the gland extirpation.

The effect of bovine interferon-alpha I1 on pregnancy rate in heifers.

Bovine interferon-alpha I1 (bIFN-alpha) may be useful for enhancing fertility in sheep and cattle because it has extensive sequence homology with ovine and bovine trophoblast protein-1 and, like those proteins, extends corpus luteum lifespan. To test the effectiveness of bIFN-alpha to enhance fertility, several experiments were performed in which inseminated heifers were given i.m. injections of bIFN-alpha approximately at the time of embryo-mediated signals that result in maintenance of the corpus luteum. In Exp. 1, heifers given 20 mg of bIFN-alpha daily from d 14 to 17 tended (P less than .07) to have lower pregnancy rates at d 110 to 112 of gestation (36/75; 48% vs 43/72; 60%). Similar results were obtained in Exp. 2 when heifers received a single injection of 40 mg of bIFN-alpha or placebo at d 13 after estrus; pregnancy rates at d 42 were 39/104 (38%) for bIFN-alpha and 47/98 (48%) for placebo. In Exp. 3, heifers were given gradually increasing doses of bIFN-alpha or placebo from d 11 to 19, because such a regimen had been shown to reduce the number of heifers experiencing hyperthermia after bIFN-alpha injection. Pregnancy rates were 42/95 (44%) for bIFN-alpha and 62/111 (56%) for placebo. Across all three experiments, pregnancy rates were lower (P less than .01) for heifers treated with bIFN-alpha (117/274; 43%) than for heifers treated with placebo (152/281; 54%). In conclusion, these results demonstrate that, under the administration systems used, bIFN-alpha does not increase pregnancy rate, but rather tends to reduce it.

Reproductive changes in male rats treated perinatally with an aromatase inhibitor.

The effects of maternal exposure to aromatase inhibitor during the perinatal period of sexual brain differentiation were studied. The fertility was assessed in adult, male rat offspring of aromatase inhibitor-treated dams. The following results were obtained: (1) Sexual maturation, body weight, and wet weights of testis, pituitary, seminal vesicle, ventral prostate, and levatori ani muscle were unchanged at adult life. (2) Fifty percent of the animals were able to mate with normal females, which became pregnant but exhibited an increased number of preimplantation loss. (3) There was a decrease in the number of spermatozoa found in the testes and in the daily sperm production. (4) Of those, 25% of the male rats treated with aromatase inhibitor did not present male sexual behavior, showing female behavior when pretreated with estrogen. These results indicate that perinatal exposure to aromatase inhibitor during the critical period of male brain sexual differentiation has a long-term effect on the reproductive physiology and behavior of male rats.

Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes

Experiments were conducted to study effects of macromolecules on stallion sperm capacitation and fertilization as determined by penetration of bovine zona-free and equine partially zona-removed oocytes. Stallion sperm were capacitated in TYH medium (modified Krebs-Ringer bicarbonate) supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/ mL of BSA. Capacitation was induced with 8 bromoadenosine cyclic monophosphate (8BrcAMP; 0.5 mM) alone or in combination with 0.1 μM of ionomycin. Intraspecies gametes were co-incubated in TYH/PVA or TYH/ BSA for 18 to 20 h. For zona-free bovine oocytes, penetration rate (35%) with the combination of 8BrcAMP and ionomycin in PVA-containing medium was higher (P < 0.05) than any treatment in BSA-containing medium (5 to 6%). A similar study was conducted using equine oocytes with partially removed zonae. Sperm capacitated and used for in vitro fertilization (IVF) in PVA-containing medium had higher penetration rates (P < 0.01) than sperm in BSA-containing medium (54 vs. 11%). The effect of equine preovulatory follicular fluid on bovine oocyte penetration was assessed. Bovine oocytes were matured in tissue culture medium-199 with 0, 20, 50, or 100% equine preovulatory follicular fluid, and 1 IU/mL of equine chorionic gonadotropin. Stallion sperm were treated with 8BrcAMP + ionomycin in PVA- or BSA-containing media. The penetration rates of bovine zona-free oocytes by stallion sperm were again higher with PVA (47%) than BSA (18%; P < 0.01). Penetration rates of oocytes matured in 100% follicular fluid were higher (P < 0.05) than for oocytes matured with 0% follicular fluid. The effects of equine follicular fluid and PVA/BSA during sperm capacitation on standard bovine IVF were examined. Culture of bovine oocytes with equine follicular fluid did not affect oocyte maturation or penetration rates after IVF. Bovine sperm capacitated with heparin in PVA-containing medium yielded lower (P < 0.05) fertilization rates than those capacitated in BSA-containing medium when incubated with both zona-intact and zona-free bovine oocytes. In summary, PVA was superior to BSA for ionophore-induced capacitation of equine sperm for penetration of zona-free bovine oocytes or partially zona-removed equine oocytes, but not for standard bovine IVF with bovine sperm. Zona-free bovine oocytes may be useful for assaying in vitro capacitation and fertilization of stallion sperm. © 2003 American Society of Animal Science. All rights reserved.