Gestational and lactational exposure to gossypol alters the testis transcriptome.

BACKGROUND: Reproductive capacity can be altered by challenges experienced during critical periods of development, including fetal development and early neonatal life. Gossypol is a polyphenolic compound, commonly found in cotton seeds, that impairs male reproduction. Here, we investigated whether the exposure to gossypol in utero and during lactation alters male reproductive function in sheep. From conception until 60 days postpartum, ewes were randomly assigned to a control diet or a gossypol-rich diet based on cottonseed. Lamb testicles were removed at 60 days of age and subjected to RNA-sequencing. RESULTS: Lambs derived from the maternal cottonseed diet showed significantly lower growth and lower testis weight as a proportion of the total body weight, and reduced testosterone levels. In addition, the testis transcriptome was significantly altered by the maternal cottonseed diet. Most of the altered genes are directly implicated in testis development and sperm biology, cell communication, iron ion metabolism, calcium homeostasis and signaling, among other functions. Interestingly, network analysis revealed that exposure to gossypol significantly disturbed coexpression patterns among spermatogenesis-related genes, suggesting a disruption in coregulation mechanisms. CONCLUSIONS: Our findings provide evidence that maternal exposure to gossypol alters male reproductive function in the offspring, with potential lasting or lifelong negative consequences.

Effects of Maternal Dietary Cottonseed on the Profile of Minerals in the Testes of the Lamb.

The use of cotton co-products in animal feed is restricted by the presence of gossypol, which is a toxic and highly reactive molecule of complex minerals. In mammals, part of the offspring phenotype is influenced by dam nutrition. The aim of this study was to investigate the effect of ewe diet, with and without cottonseed (gossypol), on the testicular development of lambs from birth to weaning through the assessment of lamb live weight (LW), macro and histological morphology of testes, and mass fraction of chemical elements by neutron activation analysis (NAA) and microprobe X-ray fluorescence spectroscopy, as well as the multielement distribution map in the testes. Eighteen lambs were used with an average LW at birth of 4 ± 1.0 kg. All lambs were offspring of Santa Inês ewes, fed on ration either with or without cottonseed during mating, gestation, and lactation, thus forming two treatments: control group (C) without cottonseed and treatment group with cottonseed (G). The animals were weighed from birth to 60 days of age, at fortnightly intervals. At 60 days of age, the lambs were orchidectomized to collect their testes for macroscopy, histological, neutron activation, and X-ray fluorescence analysis. Besides dry matter (DM), protein, ether extract (EE), calcium (Ca), and potassium (K) were higher in the ewe milk from the C group compared with the G group (P < 0.05). Lambs from the C group showed higher LW from 45 days onwards, as well as higher average daily gain when compared with the G group (P < 0.05). They also presented higher testicular weight, volume, length, width, as well as tubule and lumen diameters compared with lambs from the G treatment (P < 0.05). Multielementary NAA revealed higher mass fractions of rubidium (Rb), selenium (Se), and cesium (Cs) in the testes of lambs from the C group when compared with the G group, while G showed higher zinc (Zn) content (P < 0.05). No differences between treatments were found for element levels and distribution using X-ray fluorescence microanalysis (P > 0.05). In conclusion, the maternal cottonseed diet compromised performance and testes development of the lambs and reduce the content of Se in the testes.

Cottonseed feeding changes oxidative stress markers in ewes during the peripartum period and increases the quality of colostrum.

Immune indicators and oxidative stress markers were analyzed in blood and lacteal secretions of twenty Santa Ines ewes distributed in two feeding regimes: cottonseed concentrate (CS; n=10) and soybean concentrate (SB; n=10). Blood catalase activity and oxygen radical absorbance capacity were higher in SB than CS (P<0.05), suggesting that cottonseed increased oxidative stress in ewes. Cottonseed consumption increased IgG concentration, lysozyme activity and total protein concentration in the colostrum and decreased iron stock, suggesting a positive effect of this nutrient on the immune quality, antibacterial activity and antioxidant potential of colostrum.

Effect of maternal cottonseed feed on the immune and antioxidant status of Santa Ines lambs.

Cottonseed has been used as a nutritional alternative in animal production. However, consequences of this nutrient in the progeny is not well characterized. Thus, this work evaluated the effect on the immune and antioxidant status of the progeny of feeding Santa Ines ewe with or without cottonseed. Twenty-four Santa Ines ewes were distributed in two feeding regimes: cottonseed (CS) concentrate (n = 12) and soybean (SB) concentrate (n = 12). After birth, lambs remained with their mothers and blood samples were collected at 1st, 3rd, 7th, 15th, 30th and 60th day of life of 24 lambs born from mothers fed with (CS, n = 12) or without (SB, n = 12) cottonseed. Serum total protein, albumin, alpha beta globulin, gamma globulin, immunoglobulin G and M, activity of glutathione peroxidase (GPx), catalase (CAT), oxygen radical absorbance capacity (ORAC) and variables related to iron metabolism were affected only by sampling times (P < 0.05). The concentration of serum total protein, alpha beta globulin, gamma globulin and immunoglobulin G and M, GPx activity and ORAC values decreased as lamb age increased. Serum albumin concentration and CAT activity, in turn, increased as lamb age increased. In this work, maternal feeding with cottonseed did not affect the serum protein profile and antioxidant status of progeny during the lactation period, indicating no transfer of gossypol effects by milk secretions. Thus, the alternative in ruminants feeding with cottonseed can be used without maternal-descendant effects to immunity and oxidative stress in lambs.

Vitrification of bovine preantral follicles with dimethylsulfoxide and sucrose plus alfa-tocopherol / Vitrificação de folículos pré-antrais bovinos com dimetilxulfoxido e sacarose adicionado de alfa-tocopherol

The objective of this study was to evaluate the vitrification of bovine preantral follicles with dimethylsulfoxide (D) and sucrose (S) plus α-tocopherol 5mmol/L (T5) or 10mmol/L (T10) and, evaluate the thawed with minimal essential medium (m) with or without sucrose (s). Ovaries of cows were collected from slaughterhouse for the experiment I (n=66) and II (n=51). In the laboratory ovarian fragments were randomly assigned either to fresh control and 8 vitrification treatments (Controle and Dm; Dms, DSm; DSms; DST5m; DST5ms; DST10m; DST10ms). Ovarian fragments were placed in vitrification solution (5 min) and immersed in liquid nitrogen (-196°C), after a week, the fragments were thawed and analyzed. In the experiments I, preantral follicles were morphologically observed for histological evaluation, (normal; degenerated and developing of stage). In the experiment II, preantral follicles were mechanically isolated from ovarian tissue and examined with trypan blue, where dead and live corresponded to stained or non-stained. The treatments DSm, DSms and DST10m were effective in preserving the morphology in situ. However, the viability of isolated preantral follicles after vitrification remained high only in treatment DST10m. Thus, DST10m preserves survival rates and morphological integrity during vitrification of bovine preantral follicles.

Os objetivos deste estudo foram avaliar a vitrificação de folículos pré-antrais bovinos com dimetilsulfóxido (D) e sacarose (S) adicionando α-tocoferol 5mmol/L (T5) ou 10mmol/L (T10) e, avaliar o aquecimento com meio essencial mínimo (m) com ou sem sacarose (s). Ovários de fêmeas bovinas foram coletados de abatedouro, para o experimento I (n= 66) e II (n= 51). No laboratório fragmentos ovarianos foram distribuídos aleatoriamente para o controle fresco e 8 tratamentos de vitrificação (Controle e Dm; Dms, a DSm; DSms; DST5m; DST5ms; DST10m; DST10ms). Os fragmentos ovarianos foram colocados na solução de vitrificação (5 min) e imersos em nitrogênio líquido (-196°C). Após uma semana os fragmentos foram aquecidos e analisados. No experimento I, folículos pré-antrais foram observados morfologicamente para avaliação histológica (normal, degenerados e estádio de desenvolvimento). No experimento II, folículos pré-antrais foram mecanicamente isolados do tecido ovariano e examinados com o azul de trypan, observando mortos e vivos corados e não corados respetivamente. Os tratamentos a DSm, DSms e DST10m foram eficazes na preservação da morfologia in situ. No entanto, a viabilidade de folículos pré-antrais isolados após a vitrificação manteve-se elevada apenas no tratamento DST10m. Assim, DST10m preservou as taxas de sobrevivência e integridade morfológica durante a vitrificação de folículos pré-antrais bovinos.