Vanillic acid and vitamin C attenuated di-2-ethylhexyl phthalate-induced testicular toxicity in adult male rats.

Abstract: Di-2-ethylhexyl phthalate (DEHP) is an extensively used plasticizer which has raised some concerns about its safety on human health. This study aimed at evaluating the effects of vanillic acid (VA) and vitamin C (VC) supplementation on DEHP-induced testicular toxicity. Thirty-five adult male Wistar rats were randomly divided into 7 groups (A-G) (n = 5) receiving distilled water; 250 mg/kg bw of DEHP only; 30 mg/kg bw of VA and 250 mg/kg bw of DEHP; 30 mg/kg bw of VC and 250 mg/kg bw of DEHP; 30 mg/kg bw of DEHP plus 30 mg/kg bw of VA and 30 mg/kg bw of VC; 30 mg/kg bw of VA only; and 30 mg/kg bw of VC only, respectively. At the end of the experiment, blood was taken from the heart via cardiac puncture and stored, semen was collected from the caudal epididymis for immediate sperm analysis, while the testes were excised and preserved for histological examination and biochemical analysis. The results showed a significant decrease (P < 0.05) in body weights, sperm motility, sperm volume, sperm viability and count, antioxidant levels, and reproductive hormonal levels, with a significant increase (P < 0.05) in sperm morphological defect and lipid peroxidation level in DEHP-only group compared with the control but was ameliorated after VA and VC administration compared to the DEHP-only treated animals. VA and VC supplementation attenuated the toxic effects of DEHP on the testicular functions, morphology, and semen characterization of the experimental adult male Wistar rats. Lay summary: Male infertility is considered when identifiable female causes of infertility are excluded and semen quantity and quality fail to fulfil World Health Organization criteria. From conception through to adulthood, people are exposed to limitless environmental toxicants among which di-2-ethylhexyl phthalate (DEHP) commonly found in personal care products, cosmetics, and medical devices is prevalent. The present study elaborated on the importance of taking antioxidant-rich foods containing vitamin C and vanillic acid, such as those found in various fruits, olives, whole wheat, and cereal grains, in combating infertility caused by environmental toxicants. An experiment was carried out on rats to see the effect of vanillic acid and vitamin C supplementation on preventing DEHP-induced testicular toxicity. The testicles and semen were analyzed from five rats in each treated and control groups. The data led us to conclude that vanillic acid and vitamin C supplementation do have attenuating effects on DEHP-induced testicular toxicity, due to their high antioxidant and anti-inflammatory properties.

Effect of Zinc, Selenium, and Vitamin E Administration on Semen Quality and Fertility of Male Dromedary Camels with Impotentia Generandi.

This study aimed to investigate the effect of zinc (Zn), selenium (Se), and vitamin E (Vit E) administration on semen quality and fertility in male dromedary camels with impotentia generandi (IG, post-coital infertility). Factors that may affect response to treatment were investigated. Thirty-three IG-affected and five fertile camels were included. Case history was obtained, and breeding sound examination was performed. Semen was collected using electroejaculation. IG-camels were classified according to initial sperm count, body condition score, age, duration of infertility, IG-type, and testicular size. IG-camels were treated with a combination of intramuscular injections of Vit E (α-tocopherol acetate, 1 mg/kg bw) and Se (sodium selenite, 0.088 mg/kg bw) once every week for three successive weeks and by daily oral administration of 360 mg of zinc gluconate for 5 successive weeks. Semen quality was estimated before and after treatment. IG-treated camels were allowed to mate females in estrus, and conception rates were calculated. The results showed that sperm cell concentration, sperm motility, and viability significantly increased, while sperm abnormality significantly decreased after treatment (P < 0.01). Positive correlations were observed between initial sperm cell count and post-treated sperm count (P = 0.001), sperm motility (P = 0.01), and viability (P = 0.002). Other variables and their interactions did not affect response to treatment. Conception rates improved after treatment. In conclusion, Zn, Se, and Vit E administration improved semen quality and fertility in male dromedary camels with impotentia generandi. Initial sperm count can be used to predict the degree of camel response to treatment.

Efeito de diferentes concentrações de ingestão de gossipol livre sobre morfometria testicular e qualidade seminal de touros da raça Nelore / Effect of free gossypol intake in different levels on the testicular morphometry and seminal quality of Nelore bulls

A cotonicultura tem forte fator de impacto na economia nacional, e o estado de Mato Grosso se destaca por ser o maior produtor de algodão herbáceo e deter o maior rebanho bovino do país, condições essas que estimulam o uso do caroço, da torta e do farelo de algodão na alimentação animal. Considerando que o gossipol está presente nos subprodutos do algodão e que seus efeitos sobre a reprodução podem reduzir a fertilidade dos animais, objetivou-se, com essa pesquisa, avaliar o efeito da ingestão de dietas com diferentes teores de gossipol livre por dia sobre a morfometria testicular e a qualidade seminal de touros da raça Nelore. Foram utilizados 28 touros, distribuídos aleatoriamente em seis tratamentos: T0, 0 grama de gossipol livre/touro/dia ; T1, 1,08 grama de gossipol livre/touro/dia; T2, 2,07 gramas de gossipol livre/touro/dia; T3, 3,24 gramas de gossipol livre/touro/dia; T4, 3,82 gramas de gossipol livre/touro/dia e T5, 5,08 gramas de gossipol livre/touro/dia. Os animais de cada tratamento foram mantidos confinados em área média de 100m2, dotada de bebedouro, cochos para mistura mineral e para volumoso/concentrado. O consumo de 3,24 gramas de gossipol livre/touro/dia alterou a qualidade espermática e a morfometria testicular de touros.(AU)

The cotton industry has a strong impact factor in the Brazilian economy and the state of Mato Grosso stands out for being the largest upland-type cotton producer and also holds the largest cattle herd in the country, conditions that stimulate the use of cottonseed pie and cottonseed meal in animal feed. Whereas gossypol is present in cotton by-products and their effects on reproduction can reduce the animal fertility, this research evaluated the effect of diets with different free gossypol contents in the testicular morphometry and semen quality of Nelore bulls. 28 bulls were randomly distributed in six treatments: T0, receiving no free gossypol; T1, receiving 1.08g / bull / day; T2, receiving 2.07g / bull / day; T3, receiving 3.24g / bull / day; T4, receiving 3.82g / bull / day and T5, receiving 5.08g / bull / day of free gossypol respectively. The animals in each treatment were kept confined in an averaged area of 100m2, having fresh water, troughs for mineral mixture and roughage / concentrate. The consumption of 3.24g of free gossypol / bull / day altered the bulls sperm quality and testicular morphometry.(AU)

Impotentia generandi in male dromedary camels: heavy metal and trace element profiles and their relations to clinical findings and semen quality.

The aim of this study was to investigate the profiles of cadmium (Cd), lead (Pb), selenium (Se), zinc (Zn), copper (Cu), molybdenum (Mo), iron (Fe), and manganese (Mn) in serum of dromedary camels with impotentia generandi and their associations with the clinical findings and semen analysis data. Sixteen male dromedary camels with impotentia generandi (IG group) and 5 fertile camels (FERT group) were used. The external and internal genital organs were examined using visual inspection, palpation, and ultrasonography. Semen was collected by electroejaculation and examined for volume, count, motility, viability, and abnormality. Blood was collected from all camels and serum was harvested. All serum samples were digested by concentrated acids and analyzed for heavy metals and trace elements by flame emission atomic absorption spectrophotometer. Results showed that the mean heavy metal and trace element concentrations in serum were in the following descending order Fe > Zn > Cu > Cd > Mo > Se > Mn > Pb. Cd was higher in IG than in FERT males (P = 0.02). Se was greater in FERT than in IG groups (P = 0.003). Zn was higher in in FERT than in IG groups (P = 0.001). There were positive correlations between Zn and sperm count (r = 0.59, P = 0.005) and sperm motility (r = 0.57, P = 0.005) and a tendency for negative correlation between Zn and sperm abnormalities (r = - 0.44, P = 0.05). In conclusion, Cd might be implicated as a cause of infertility in male camels. Deficiencies of Se and Zn may also have adverse impacts on male camel reproduction.

Selenium and vitamin E supplementation enhances the antioxidant status of spermatozoa and improves semen quality in male dogs with lowered fertility.

Studies showed a beneficial effect of supplementation with selenium (Se) and vitamin E on semen quality. The aim of the study was to investigate the effect of Se and vitamin E supplementation on the antioxidant status of spermatozoa and semen quality in dogs with lowered fertility. Ten dogs were supplemented daily with Se (6 µg/kg organic Se yeast) and vitamin E (5 mg/kg) per os for 60 days. Control group consisted of 10 males without the supplementation. Semen was collected on day 0, 30, 60 and 90. Sperm quality parameters were evaluated using CASA and a microscope. Concentrations of Se and vitamin E in blood as well as glutathione peroxidase (GSH-Px) activity and total antioxidant capacity (TAC) in the spermatozoa were determined. After 60 days of supplementation the concentration of spermatozoa, the majority of motility indicators and the percentage of normal morphology and live spermatozoa increased significantly (p < .05). An increase (p < .05) in concentration of Se and vitamin E in blood and GSH-Px-activity and TAC in the spermatozoa was detected. The study results indicate that Se and vitamin E supplementation for 60 days enhances the antioxidant status of spermatozoa and improves the quality of the semen in dogs with lowered fertility.

Increased sperm cell production in ageing roosters by an oral treatment with an aromatase inhibitor and a natural herbal extract designed for improving fertility.

Rooster fertility peaks between 30 and 40 weeks of age and declines rapidly from 50 weeks of age. This is linked to a reduction in the number of spermatozoa in the ejaculate due to high density of spermatids in the seminiferous tubules of low-fertility roosters. In this study, we assessed the effects over spermatogenesis of both letrozole, an aromatase inhibitor, and a commercial herbal extract designed for improving fertility composed by Tribulus, Cinnamomum, Zingiber and Sativus. Forty-two-week-old Ross 308 roosters (n = 24) were distributed into four groups: control, letrozole (0.03 mg/kg), herbal extract (0.04 ml/kg) and letrozole+herbal extract. After 14 weeks of daily oral supplementation, their testes and epididymides were weighed, fixed and sectioned for assessment of spermatogenesis and quantification of sperm cells inside the lumen. Differences in seminiferous tubules measurements and density of sperm cells were tested using R software (version 3.0.1). Although body weight was not affected by the treatment, testes from animals treated with the combination of letrozole and herbal extracts were heavier than those from control animals. Animals treated with either letrozole or herbal extract, or their combination, showed a significant higher number of sperm cells inside the seminiferous tubules and epididymis than control animals (p < .05). These data suggest that the use of letrozole and the herbal extract could improve sperm cell production in ageing roosters. Future studies are needed to disclose the causal mechanisms involved and its effect on fertility and ejaculate features.

The effect of individualized homeopathic treatment on the semen quality of bulls with reproductive disorders: a case series.

BACKGROUND: Poor semen quality of pedigree bulls has major economic implications in cattle breeding. AIMS: To evaluate the effect of homeopathy on the semen of bulls with reproductive disorders. METHODS: The behavioral, clinical and spermatic characteristics of four Nelore bulls were evaluated. The bulls received individualized homeopathic treatment mixed into the feed and administered once per day. Semen was collected using an artificial vagina. Successful collection, freezing, effective doses, and the number of doses of semen per collection were compared before and after homeopathic treatment. RESULTS: The bulls were treated with Sulphur, Phosphorus, Lycopodium and Argentum nitricum all in 30CH dilution. The rates of successful collection were 97.14%, 100%, 96.67% and 30.77% pretreatment and 95.45%, 100%, 94.67% and 96.77% at post-treatment for bulls A, B, C and D, respectively. The average number of doses per ejaculate pre and post-treatment were 102.67 (SD 74.41) vs. 300.08 (SD 180.58), 0 vs. 234.78 (SD 96.12), 0 vs. 105.12 (SD 54.98), 0 ± vs. 107.37 (SD 52.12) respectively. Many of these differences were statistically significant. CONCLUSION: The use of homeopathy apparently improved the production of viable doses of semen from bulls with previous freezing problems and poor semen quality. Controlled studies should be conducted.

Alleviation of reproductive toxicity of gossypol using selenium supplementation in rams.

The objective of this study was to evaluate the reproductive toxicity of gossypol and the possible counteracting effect of selenium supplementation in rams. Twenty-five mature crossbred (Barki × Rahamni) rams were randomly divided into 5 equal groups. The first group served as a control (no gossypol in diet). The second and third groups received diets containing free gossypol of 9 and 14 mg·kg(-1) BW·d(-1)(low level and high level of gossypol), respectively. The fourth and fifth groups received the same diets given to the second and third groups, respectively, with an oral administration of 1 mg selenium (as sodium selenite) daily for each animal. Results showed reductions in ejaculate volume (P = 0.028), percentage of dead sperm (P = 0.003), total functional sperm fraction (P < 0.001), and blood serum concentration of testosterone (P < 0.001) in the presence of both levels of gossypol inclusion. Furthermore, high level of gossypol reduced forward motility (P < 0.001) and semen initial fructose concentration (P = 0.002) and increased abnormal-head sperm (P = 0.003) and blood serum concentration of triiodothyronine (P = 0.006). Regardless of selenium supplementation, increasing level of free gossypol in diet resulted in a significant decrease in the percentage of forward motility (P = 0.037) and significant increases in the mean values of sperm concentration (P < 0.001), total sperm output (P = 0.002), percentage of total abnormal sperm (P = 0.058), and abnormal-head sperm (P = 0.016). On the other hand, regardless of levels of gossypol inclusion, selenium supplementation resulted in significant increases in libido (P < 0.001), mean values of ejaculate volume (P < 0.001), percentage of forward motility (P = 0.019), total sperm output (P < 0.001), total functional sperm fraction (P < 0.001), semen initial fructose concentration (P = 0.031), and blood serum concentrations of both testosterone and triiodothyronine (P < 0.001). In conclusion, free gossypol in diet induced adverse effects on semen quality in rams, but selenium supplementation successfully counteracts most of the hazardous effects of gossypol on semen characteristics.

Evaluation of sperm DNA peroxidation in fertile and subfertile dogs.

Oxidative stress (OS) has been recognized as one of the most important causes of male infertility. The antioxidant activities of seminal plasma and epididymal fluid are not enough to prevent OS, which can damage sperm membranes and DNA, so antioxidant supplementation has been used as a treatment of male infertility. The aim of this experiment was to evaluate the DNA peroxidation before and after antioxidant supplementation with vitamin C and E in dogs with and without fertility problems. A total of eleven dogs were used and were divided in two groups: fertile group (G1), dogs with normal spermiogram (n = 5); subfertile group (G2): dogs with low sperm count (<20 × 10(6) sptz/ml) and/or more than 30% of total sperm pathology (n = 6). Both groups received 500 mg/day of vitamin C and 500 mg/day of vitamin E for 60 days. A semen sample was collected before (M1) and after (M2) oral supplementation. Samples were analysed for DNA peroxidation by measuring the 8-hydroxy-2'-deoxyguanosine concentration. No significant difference was observed between groups at either time. Oral supplementation with 500 mg/day of vitamin C and 500 mg/day of vitamin E did not change the DNA peroxidation in fertile and subfertile dogs.

Sperm morphological studies of the west African dwarf buck treated with pumpkin plant (Cucurbita pepo) / Estudio morfológico del esperma de la cabra enana del oeste africano tratado con planta de calabazas (Cucurbita pepo)

Four healthy bucks of the West African Dwarf breed aged between 24 and 30 months were used for this study. The bucks were first used as control and later as experimental animals upon being fed daily with oiled pumpkin plant for the period of six months. The objective of the study was to investigate the effect of the pumpkin plant on the morphology of the spermatozoa of the bucks. There were significant differences (p< 0.05) between the control and experimental values for both primary and secondary morphological spermatozoa abnormalities: the pyriform head has a control value of 6 (0. 42 percent) and post-feeding value of 0 (0 percent), the beat tail; 14(0.97 percent) and 2 (0.16 percent) for the control and post feeding values respectively. The curved mid piece: 17 (1.18 percent) and 1 (0.08 percent) for the control and post-feeding values respectively. The bent mid piece also differed significantly (p<0.05) between control value of 16 (1.11 percent) and post feeding value of 3 (0.23 percent). All through the stages of the study, there were significant reductions in the number of sperm cells with abnormalities consequent upon daily feeding of the animals with pumpkin plant. The plant is therefore recommended for both prevention and treatment of make infertility in man and animals.

Cuatro cabras de la raza enana del oeste de África, con edades comprendidas entre los 24 y 30 meses, fueron utilizadas para este estudio. Las cabras sirvieron primero como control y, posteriormente, como animales de experimentación los que se alimentaron diariamente con aceite de planta de calabaza por un período de seis meses. El objetivo del estudio fue investigar el efecto de la planta de calabaza sobre la morfología de los espermatozoides de las cabras. Hubo diferencias significativas (p <0,05) entre los valores control y experimental en las anomalías morfológicas primarias y secundarias de los espermatozoides: la cabeza tuvo un valor de control de 6 (0,42 por ciento) y post alimentación valor 0 (0 por ciento), el movimiento de la cola un valor de 14 (0,97 por ciento) y 2 (0,16 por ciento) para el control y post alimentación, respectivamente. La curvatura media: valores de 17 (1,18 por ciento) y 1 (0,08 por ciento) para el control y post-alimentación, respectivamente. La inclinación media también difería significativamente (p <0,05), con un valor entre 16 (1,11 por ciento) el control y 3 (0,23 por ciento) post-alimentación. Hasta el final de las etapas del estudio, hubo una reducción significativa en el número de células de esperma con anomalías como consecuencia de la alimentación diaria de los animales con la planta de calabaza. Por lo tanto, la planta se recomienda para la prevención y el tratamiento de la infertilidad en hombres y otros animales.

Improvement of sperm production in subfertile boars by Cordyceps militaris supplement.

Cordyceps species have been traditionally used for the enhancement of sexual function, however, there is few direct evidence to prove this. We investigated the spermatogenic effect of Cordyceps militaris (CM) by supplementation with CM mycelium to subfertile boars. Seventeen Duroc and 12 Landrace boars (29 to 40 months old) were selected to feed with regular diet (control groups, n = 8 and 6, respectively) or diet supplemented with CM mycelium (treatment groups, n = 9 and 6, respectively) for 2 months. Semen was collected once a week. The quality of fertile sperm (normally greater than 62% of motility and 70% of normal morphology) and the quantity (semen volume, and total sperm number) were compared in these boars. The result showed that sperm production was enhanced significantly at the end of first month (p < 0.05), peaked at the second month (p < 0.01) of supplementation with CM and was maintained for 2 weeks after stopping the treatment (p < 0.01). Plasma cordycepin concentration was detected in boars supplemented with CM but not in the controls. More importantly, the percentages of motile sperm cells and sperm morphology were also improved significantly in most of treated boars during the second month of supplementation (p < 0.01) and 2 weeks after the treatment (p < 0.05) as compared to their initial values. These results indicate that supplementation with CM mycelium improves sperm quality and quantity in subfertile boars and may partly support the role of Cordyceps in sexual enhancement.

Homeopathic treatment for infertility in a prize Nelore bull.

Treatments for infertility in bulls are not described in homeopathic literature. A few treatments, such as changing the protein content of the diet, giving extra minerals, etc have been proposed. This case report describes homeopathic treatment for infertility in a prize bull. A Nelore bull, considered infertile for 3 years, was treated with homeopathic Pulsatilla nigricans 200 CH. Decreased total sperm defects, increased sperm motility and a very impressive increased number of doses of semen produced were observed. The bull relapsed after treatment was withdrawn, but again responded when it was resumed. Since only one animal was observed one cannot assume that the observed changes were due only to this treatment. Further studies may establish the real benefits of a homeopathic medicine in bull infertility.

Trace element deficiencies and fertility in ruminants: a review.

Various minerals (copper, cobalt, selenium, manganese, iodine, zinc, and iron) can influence reproductive performance of ruminants. Reproductive failure may be induced by deficiencies of single or combined trace elements and by imbalances. This review is focused on maladjustments of trace elements leading to impaired breeding performance. Opinion is diverse as to the existence of various reproductive disturbances from either a severe copper depletion or a marginal dietary copper deficiency. Field experience suggests that administration of cobalt to ruminants on cobalt-deficient diets improves their impaired breeding performance. Selenium infertility in ewes is more prevalent in some areas and in some seasons, but the actual cause of this malady and the continuing role of additional factors are unknown. Manganese is necessary for normal fertility in ruminants, and feeding low-manganese rations depresses conception rates. Lack of iodine impairs thyroid activity and also ovarian function. Reproductive failure in the female and in spermatogenesis are manifestations of zinc deficiency. Despite forages rich in iron, low availability in certain instances could affect adversely ruminant reproduction. Knowledge of biochemical dysfunctions from trace element deficiencies is essential to determine the role which trace elements play in fertility of ruminant animals.

Histological observation of some of the endocrine glands in the sterile carp-funa hybrid (F1), with special reference to the hypophysis.

Some endocrine glands of the carp-funa hybrids were studied with a light microscope to elucidate their detailed structure and the possible causal factor of sterility in the males. Adult specimens of carp (Cyprinus carpio), gengoroh-buna (Carassius auratus cuvieri), and their hybrid (F1) were examined. The hybrid males are sterile as manifested by the failure of meiosis and seminomatous neoplasm in their testes. The hybrid females revealed well-developed ovaries, but their fertility was not tested. The hybrid hypophysis shows an intermediate condition between the parent species in the grade of ramification of the pars nervosa into the pars intermedia. Among seven types of granular cells demonstrated in the adenohypophysis, certain degenerative and anomalous changes are recognized only in the gonadotrophs of the hybrid hypophysis, especially in the female. These changes are discussed as a possible cause of sterility. A considerable amount of aldehyde fuchsin stainable neurosecretory material occurs in the cells of the nucleus preopticus and in the pars nervosa. The nucleus lateralis tuberis exhibits a histologically healthy condition.