Effects of high temperature and LPS injections on the hemocytes of the crab Neohelice granulata.

Temperature fluctuations, particularly elevated temperatures, can significantly affect immune responses. These fluctuations can influence the immune system and alter its response to infection signals, such as lipopolysaccharide (LPS). Therefore, this study was designed to investigate how high temperatures and LPS injections collectively influence the immune system of the crab Neohelice granulata. Two groups were exposed to 20 °C (control) or 33 °C for four days. Subsequently, half were injected with 10 µL of physiological crustacean (PS), while the rest received 10 µL of LPS [0.1 mg.kg-1]. After 30 min, the hemolymph samples were collected. Hemocytes were then isolated and assessed for various parameters using flow cytometry, including cell integrity, DNA fragmentation, total hemocyte count (THC), differential hemocyte count (DHC), reactive oxygen species (ROS) level, lipid peroxidation (LPO), and phagocytosis. Results showed lower cell viability at 20 °C, with more DNA damage in the same LPS-injected animals. There was no significant difference in THC, but DHC indicated a decrease in hyaline cells (HC) at 20 °C following LPS administration. In granular cells (GC), an increase was observed after both PS and LPS were injected at the same temperature. In semi-granular cells (SGC), there was a decrease at 20 °C with the injection of LPS, while at a temperature of 33 °C, the SGC there was a decrease only in SGC injected with LPS. Crabs injected with PS and LPS at 20 °C exhibited higher levels of ROS in GC and SGC, while at 33 °C, the increase was observed only in GC and SGC cells injected with LPS. A significant increase in LPO was observed only in SGC cells injected with PS and LPS at 20 °C and 33 °C. Phagocytosis decreased in animals at 20 °C with both injections and exposed to 33 °C only in those injected with LPS. These results suggest that elevated temperatures induce changes in immune system parameters and attenuate the immune responses triggered by LPS.

Probe ultrasonification of egg yolk plasma forms low-density lipoprotein nanoparticles that efficiently protect canine semen during cryofreezing.

Around the world, many couples have turned to in vitro fertilization as a viable solution to fertility issues. Low-density lipoprotein (LDL) is a protein best known for transporting fat molecules throughout the body, but it has also been shown to protect sperm cells during cryopreservation due to its micellar structure. In the present study, we aimed to evaluate different protocols for the preparation of nanoparticles from egg yolk plasma (EYP) containing LDL to improve the viability of cryopreserved canine semen. EYP was subjected to three distinct treatments: ultrasonification in an ultrasound bath at 40 kHz for 30 min (LDL-B); ultrasonification via an ultrasound probe at 50% amplitude for 30 min (LDL-P); and high-pressure homogenization at 10,000 PSI for six cycles (LDL-H). Sperm quality was assessed after thawing using computer-assisted sperm analysis and flow cytometry. The results revealed that compared to the EYP control, the LDL-P formulation presented significantly higher efficiency (p < 0.05) in maintaining total and progressive sperm motility, sperm membrane integrity and fluidity, and levels of intracellular reactive oxygen species. The LDL-P nanoparticles had an average size of approximately 250 nm, a PDI value of 0.3, and -1.15 mV of zeta potential, which are very important because it is an indicator of the stability of a colloidal dispersion. Therefore, we conclude that ultrasonication of EYP using a probe is an efficient method for the preparation of LDL nanoparticles that would enhance the cryoprotection of semen during freezing.

Acute exposure to isoflurane impairs sperm parameters in mice.

Isoflurane, an inhalational anesthetic from the halogenated group, has been increasingly used in the medical and scientific fields. Due to its characteristics, it is capable of inducing anesthesia quickly and quietly; however, the adverse effects resulting from its use have not yet been fully elucidated, especially with regard to reproductive aspects. Considering its common use in research laboratories, whether for performing surgical procedures or for prior exposure to euthanasia, knowledge about its interference in sperm parameters of experimental models characterizes an important study goal. The aim of the present study was to determine the interference of acute exposure to isoflurane on the sperm quality of mice, both immediately previous to euthanasia and in later evaluation, twenty days after a single anesthetic exposure. Our results demonstrate that acute anesthetic exposure reduces sperm motility and is responsible for the formation of damaged sperm cells that are prone to apoptosis, which may affect the outcome of reproductive experiments even 20 days after exposure.

Nanocarried antioxidants in freezing extenders for boar spermatozoa.

Post-thawing cryoinjuries in boar spermatozoa due to oxidative stress may be reduced by adding nanoencapsulated antioxidants to freezing extenders. This study evaluated post-thawing kinetics, structural and biochemical functions of boar spermatozoa frozen with extenders including resveratrol and vitamin E loaded into polymeric nanocapsules. Resveratrol was added at 0 (control), 5, 10, 20, 40 and 80 µg/ml, whereas Vitamin E was added at 0 (control), 50, 100, 200 and 400 µg/ml. Both antioxidants were tested in free and nanoencapsulated presentations. In contact with empty nanocapsules, some sperm kinetics parameters were impaired compared to the control (p < .05), whereas lipoperoxidation declined (p < .05). With inclusion of 40 µg/ml nanoencapsulated resveratrol, some sperm kinetics parameters were improved (p < .01), but sperm motility, structural and biochemical functions did not differ from the control (p > .05). No improvement in sperm quality occurred with inclusion of vitamin E, although sperm kinetics with 400 µg/ml nanoencapsulated vitamin E was reduced compared to the control (p < .01). Inclusion of 40 µg/ml nanoencapsulated resveratrol benefitted boar sperm kinetics after thawing, but no improvement resulted from inclusion of vitamin E.

Impacts of the biocide chlorothalonil on biomarkers of oxidative stress, genotoxicity, and sperm quality in guppy Poecilia vivipara.

Chlorothalonil is a fungicide present in antifouling paints and other formulations used in agriculture, although studies have shown this chemical to be toxic to fish species. To clarify the deleterious effects of chlorothalonil for these non-target organisms, the present study evaluated the toxic effects of this biocide for the estuarine guppy Poecilia vivipara in terms of an acute mortality test (96 h) and the analysis of biomarkers of oxidative stress, genotoxicity, and sperm quality. The LC50 calculated for P. vivipara was 40.8 µg/L of chlorothalonil. For the analysis of biomarkers, fish were exposed (96 h) to 1 and 10 µg/L of chlorothalonil. It was observed that chlorothalonil alters the levels of pro- and antioxidants towards oxidative stress. In the gills, a negative effect on total antioxidant capacity (ACAP) was detected, while there was a reduction in the activity of glutathione S-transferase (GST) in the liver. However, levels of glutathione (GSH) and the activity and glutamate-cysteine-ligase (GCL) increased in both tissues, as a possible detoxification response. Following chlorothalonil exposure, oxidative damage measured by lipoperoxidation (LPO) significantly increased at the cellular level only (red blood cells (RBCs) and sperm cells). An increase in fluidity of membranes, reactive oxygen species concentration and micronuclei (MNs) incidence were also seen in RBCs. In sperm cells, LPO increased, while membrane and mitochondrial functionality as well as sperm motility decreased. Based on these results, chlorothalonil can be considered as a toxic compound for fish, causing genotoxicity and affecting the RBCs physiology and the fertility of males of P. vivipara.

Roundup® Herbicide Decreases Quality Parameters of Spermatozoa of Silversides Odontesthes Humensis.

The silverside (Odontesthes humensis) is a very interesting model for toxicological studies due its high sensitivity and need for good water quality. The aim of this study was to evaluate the effects of Roundup on spermatozoa of O. humensis, after acute exposure. The fish were exposed to 0 and 7.8 mg L-1 (a.e.) of glyphosate, respectively. Through computer-assisted sperm analysis, a significant decrease in concentration, total and progressive motility, average path distance, straight line distance, path average velocity, curved line velocity, straight line velocity linearity, wobble, amplitude of lateral head displacement, cross beat frequency, and motility period of silverside spermatozoa exposed to Roundup was observed. Also, increase in membrane fluidity, ROS production and lipid peroxidation and a decrease in the mitochondrial functionality was observed in spermatozoa of Roundup exposed silversides. It was demonstrated that Roundup exposure in a concentration that can be achieve in natural water bodies soon after its application in fields is able to cause losses in several sperm quality parameters, consequently decreasing the fertilization potential of O. humensis spermatozoa.

Effects of dietary microencapsulated Cymbopogon flexuosus essential oil on reproductive-related parameters in male Rhamdia quelen.

In aquaculture, nutrition and supplemented diets have been shown to affect broodstock reproductive performance. In this study, we investigated the effects of dietary supplementation with Cymbopogon flexuosus essential oil (CFEO) microcapsules on reproductive-related parameters in silver catfish (Rhamdia quelen) male broodfish. Adult male broodstocks were separated into three groups according to the concentrations of supplemented CFEO (0.0 = control; 1.0 or 3.0 mL per kg of diet). After 20 days under experimental conditions, the animals were euthanized and the gonads were harvested for gonadosomatic index, sperm analysis, oxidative stress, and histopathology; testosterone levels were measured in the plasma; gene expression of prl, smtl, pomca, and pomcb was assessed in the pituitary gland by real-time PCR. The results showed no alterations on reproductive parameters in R. quelen males treated with Cymbopogon flexuosus essential oil compared to the control-diet animals. In conclusion, CFEO microcapsules supplied for 20 days in the concentrations of 1.00 or 3.00 mL per kilogram of diet did not affect the reproduction criteria evaluated in this study in male silver catfish.

Effects of Experimental Lead Exposure on Testis of the Chestnut Capped Blackbird Chrysomus ruficapillus.

Lead (Pb) effects on testis histology, as well as sperm quality and oxidative status were evaluated in male Chestnut Capped Blackbird (Chrysomus ruficapillus). Wild blackbirds were captured, immediately sampled (field group) or kept in captivity and treated with a single intraperitoneal injection of saline solution (control) or saline solution with Pb acetate (50 or 100 mg/kg Pb). Seven days after injection, whole blood, ductus deferens and testis samples were collected. Increased Pb concentrations were observed in whole blood and testis of Pb-exposed blackbirds with respect to those from field and control blackbirds. Sperm cells of Pb-exposed blackbirds showed loss of membrane integrity, mitochondrial functionality, and DNA integrity. Also, oxidative damage was observed in testis of blackbirds injected with 100 mg/kg Pb. These findings indicate that Pb is accumulated in testis of C. ruficapillus, inducing severe morphological and biochemical injury that can compromise the reproductive performance of male blackbirds. Although the exposure scenario (Pb acetate, high dosage and intraperitoneal injection) tested in the present study would likely not occur in the wild, it was adequate to show potential and relevant toxic effects of Pb in wild birds.

Reproductive parameters of double transgenic zebrafish (Danio rerio) males overexpressing both the growth hormone (GH) and its receptor (GHR).

Growth hormone (GH) transgenesis presents a high potential application in aquaculture. However, excess GH may have serious consequences due to pleiotropic actions. In order to study these effects in zebrafish (Danio rerio), two transgenic lines were developed. The first expresses GH ubiquitously and constitutively (F0104 line), while the second expresses the GH receptor in a muscle-specific manner (Myo-GHR line). Results from the F0104 line showed accelerated growth but increased reproductive difficulties, while Myo-GHR did not show the expected increase in muscle mass. Since the two lines appeared to display complementary characteristics, a double transgenic (GH/GHR) was created via crossing between them. This double transgenic displayed accelerated growth, however reproductive parameters remained uncertain. The objective of the present study was to determine the reproductive capacity of males of this new line, by evaluating sperm parameters, expression of spermatogenesis-related genes, and reproductive tests. Double transgenics showed a strong recovery in almost all sperm parameters analyzed when compared to the F0104 line. Gene expression analyses revealed that Anti-Müllerian Hormone gene (amh) appeared to be primarily responsible for this recovery. Reproductive tests showed that double transgenic males did not differ from non-transgenics. It is possible that GHR excess in the muscle tissues of double transgenics may have contributed to lower circulating GH levels and thus reduced the negative effects of this hormone with respect to reproduction. Therefore, it is clear that GH-transgenesis technology should take into account the need to obtain adequate levels of circulating hormone in order to achieve maximum growth with minimal negative side effects.

Flow cytometric sex sorting affects CD4 membrane distribution and binding of exogenous DNA on bovine sperm cells.

Bovine sex-sorted sperm have been commercialized and successfully used for the production of transgenic embryos of the desired sex through the sperm-mediated gene transfer (SMGT) technique. However, sex-sorted sperm show a reduced ability to internalize exogenous DNA. The interaction between sperm cells and the exogenous DNA has been reported in other species to be a CD4-like molecule-dependent process. The flow cytometry-based sex-sorting process subjects the spermatozoa to different stresses causing changes in the cell membrane. The aim of this study was to elucidate the relationship between the redistribution of CD4-like molecules and binding of exogenous DNA to sex-sorted bovine sperm. In the first set of experiments, the membrane phospholipid disorder and the redistribution of the CD4 were evaluated. The second set of experiments was conducted to investigate the effect of CD4 redistribution on the mechanism of binding of exogenous DNA to sperm cells and the efficiency of lipofection in sex-sorted bovine sperm. Sex-sorting procedure increased the membrane phospholipid disorder and induced the redistribution of CD4-like molecules. Both X-sorted and Y-sorted sperm had decreased DNA bound to membrane in comparison with the unsorted sperm; however, the binding of the exogenous DNA was significantly increased with the addition of liposomes. Moreover, we demonstrated that the number of sperm-bound exogenous DNA was decreased when these cells were preincubated with anti-bovine CD4 monoclonal antibody, supporting our hypothesis that CD4-like molecules indeed play a crucial role in the process of exogenous DNA/bovine sperm cells interaction.

Maternal Transfer of Bisphenol A During Nursing Causes Sperm Impairment in Male Offspring.

The health effects of environmental chemicals on animals and humans are of growing concern. Human epidemiological and animal study data indicate that reproductive disorders and diseases begin early during prenatal and postnatal development. An increase of human male reproductive disturbance in the past several decades was associated to chemicals called endocrine disruptors (ED). Bisphenol A (BPA) is a ubiquitous organic environmental contaminant with ED activity. This study verified the effect of BPA exposure via breast milk during the lactation (early postnatal) period in male mice. Dams were exposed to oral BPA (300, 900, and 3000 µg/kg/BW/day) during the breastfeeding period (21 days). BPA at all concentrations significantly impaired sperm parameters in adult mice (8 months old), but mitochondrial functionality was more affected at BPA 3000. The acrosome membrane parameter was affected by BPA concentrations from 900 to 3000, and DNA integrity showed pronounced impairment at BPA 900 and 3000. BPA 3000 treatment also induced testicular degeneration and complete aplasia in some seminiferous tubules. Testicular oxidative damage was observed, and the total antioxidant capacity was impaired in BPA 900 and 3000 treatment groups. Taken together, the present study demonstrated long-term adverse effects of BPA in male mice, including reduced sperm quality, antioxidant capacity, and changes in testicular tissue. Our results clearly demonstrate the danger of BPA transferred via lactation on sperm quality registered even after a long time-elapsed from exposure to this harmful chemical.

The effects of osmolality on sperm quality in Jenynsia multidentata (Cyprinodontiformes: Anablepidae).

Sperm quality tests on fish are classically used for evaluating cryopreservation procedures, and they are also promising to assess aquatic toxicity and biomarkers of xenobiotic effects on reproduction. Osmotic shock from the storage medium is one of the main factors affecting sperm quality during evaluation. Thus, the objective of this study was to evaluate the effects of different osmolalities (240-460 mOsm/kg) for at least 4 days on the sperm quality parameters of the viviparous fish Jenynsia multidentata. The level of significance was (P < 0.05). The plasma osmolality of J. multidentata is 326 ± 3.9 mOsm/kg. The motility of fresh semen was higher in osmolalities of 280 and 300 mOsm/kg but did not differ between osmolalities from 240 to 320 mOsm/kg. Above 380 mOsm/kg, the motility observed was 0%. Over the time period studied motility increased with increasing osmolality, and the most constant and long-lasting rates were between 300 and 320 mOsm/kg. On the 4th day of evaluation, higher membrane integrity rates were observed between 280 and 360 mOsm/kg, higher mitochondrial membrane potential was observed between 300 and 460 mOsm/kg, and higher DNA integrity rates were observed between 260 and 380 mOsm/kg. Moreover, osmolalities ≥460 and ≤240 resulted in the lowest motility and DNA integrity levels. Over 4 days, the plasma membrane integrity was significantly lower at ≤260 and ≥400 mOsm/kg, and the mitochondrial membrane potential was significantly lower only in osmolalities ≤240 mOsm/kg. Therefore, we conclude that for sperm quality preservation in J. multidentata, an osmolality of 300-320 mOsm/kg of the most suitable diluent is necessary. Furthermore, we conclude that the storage of sperm in a hyposmotic (<260 mOsm/kg) or hyperosmotic (>400 mOsm/kg) solution affects not only motility but also other sperm quality parameters.

Vitrification of equine oocytes with a polyvinyl alcohol after in vitro maturation with equine growth hormone and insulin-like growth factor-I.

BACKGROUND: In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods. OBJECTIVE: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA). METHODS: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were exposed to 1.4M DMSO+1.8M EG+1 percent PVA for 3min, and then transfer to 2.8M DMSO+3.6M EG+0.6M sucrose+1 % PVA for 1min. After rewarming, oocytes were evaluated by confocal microscopy. RESULTS: Maturation rates of oocytes were not significant different among groups (P > 0.05), however eGH+IGF-I group can develop the assessment of resumption of meiosis (MI+MII = 86.7 %). CONCLUSION: The oocyte did not show morphological alterations. The use of PVA-copolymer may represent a potential alternative for vitrification of equine oocytes after IVM.

Chlorothalonil as a potential endocrine disruptor in male zebrafish (Danio rerio): Impact on the hypothalamus-pituitary-gonad axis and sperm quality.

Chlorothalonil is a broad-spectrum organochlorine fungicide widely employed in agriculture to control fungal foliar diseases. This fungicide enters aquatic environments through the leaching process, leading to toxicity in non-target organisms. Organic contaminants can impact organism reproduction as they have the potential to interact with the neuroendocrine system. Although there are reports of toxic effects of chlorothalonil, information regarding its impact on reproduction is limited. The aim of the present study was to evaluate the influence of chlorothalonil on male reproductive physiology using the zebrafish (Danio rerio) as ecotoxicological model. Zebrafish were exposed for 7 days to two concentrations of chlorothalonil (0.1 and 10 µg/L) along with a control group (with DMSO - 0.001%). Gene expression of hypothalamus-pituitary-gonad axis components (gnrh2, gnrh3, lhr, fshr, star, hsd17b1, hsd17b3, and cyp19a1), as well as hepatic vitellogenin concentration were assessed. In sperm cells, reactive oxygen species (ROS) content, lipid peroxidation (LPO), mitochondrial functionality, and membrane integrity and fluidity were evaluated. Results indicate that exposure to the higher concentration of chlorothalonil led to a reduction in brain gnr2 expression. In gonads, mRNA levels of lhr, star, and hsd17b1 were decreased at both chlorothalonil concentrations tested. Similarly, hepatic vitellogenin concentration was reduced. Regarding sperm cells, a decreased ROS level was observed, without significant difference in LPO level. Additionally, a higher mitochondrial potential and lower membrane fluidity were observed in zebrafish exposed to chlorothalonil. These findings demonstrate that chlorothalonil acts as an endocrine disruptor, influencing reproductive control mechanisms, as evidenced by changes in expression of genes HPG axis, as well as hepatic vitellogenin concentration. Furthermore, our findings reveal that exposure to this contaminant may compromise the reproductive success of the species, as it affected sperm quality parameters.

Assessment of oxidative stress biomarkers in the threatened annual killifish Austrolebias charrua exposed to Roundup.

This study aimed to analyze the toxic effects of Roundup Transorb® on the endangered Neotropical annual killifish Austrolebias charrua through the assessment of molecular and biochemical biomarkers. The fish were collected in temporary ponds and exposed to environmentally realistic concentrations of the herbicide (5 mg.L-1 for 96 h). The production of ROS, lipid peroxidation, DNA damage, and membrane fluidity were evaluated in the blood cells by flow cytometry. The mRNA expression of the antioxidant-related genes sod2, cat, gstα, atp1a1, gclc, and ucp1 across the brain, liver, and gills was quantified. The acute exposure of annual killifish to Roundup significantly increased ROS production, lipid peroxidation, and DNA damage in their erythrocytes. Likewise, Roundup Transorb® decreased membrane fluidity in the blood cells of the exposed fish. Gene expression analysis revealed that Roundup exposure alters the relative expression of genes associated with oxidative stress and antioxidant defense. Our results give rise to new insights into adaptive mechanisms of A. charrua in response to Roundup. Since Brazilian annual killifishes strongly risk extinction, this study paves the way for developing novel biotechnologies applied to environmental monitoring and aquatic toxicology assessment.

Birthweight leads to seminal and testicular morphofunctional commitment in sexually mature boars.

Given the importance of males as semen donors for artificial insemination (AI) and the high incidence of low birthweight piglets at commercial farms, the impact of birthweight on fertility in boars deserves special attention. The aim of this study was to evaluate testicular morphofunctional parameters and semen characteristics in different birthweight boars. Forty littermate males were selected at birth and divided into two experimental groups, according to birthweight: high (HW, birthweight ranging from 1.80 to 2.15 kg, n = 20) and low (LW, birthweight ranging from 0.75 to 1.10 kg, n = 20). At 170 days of age, a sub-set of 24 littermate boars (n = 12 HW and n = 12 LW) were randomly selected for semen collection, which was performed once a week, at a 15-day interval, during five weeks. At 300 days of age, boars were orchiectomized, and the testis processed for histological and molecular analyses. The HW group was heavier and presented larger testes compared to LW animals (P < 0.05). Despite that, birthweight did not significantly affect semen volume or sperm quality parameters (concentration, motility, vigor or morphology), although LW boars produced 38.2% fewer total sperm and 24% lower semen concentration, leading to 36.8% less inseminating doses. The histomorphometrical evaluation showed that seminiferous tubules diameter and germinal epithelium height were similar between experimental groups. However, LW boars presented shorter seminiferous tubules and, consequently, fewer Sertoli cells per testis (P 0.05). Even though plasma testosterone levels were equivalent in both birthweight groups, LW testis presented less androgen receptors (P < 0.05). Additionally, birthweight was positively correlated with total seminiferous tubule length and number of Sertoli cells (P < 0.01), and with body and testis weights (P < 0.01). Taken together, even though adult LW boars showed no evident semen pathologies or spermatogenesis commitment, mature HW males have the potential to produce more spermatozoa, consequently more semen doses per ejaculate, being more valuable to an industry that relies on AI.

Oral cytological changes in young adults related to alcohol consumption.

OBJECTIVE: The study aimed to investigate the oral cytological changes in young adults with recent history of alcohol consumption, as well as its relation with the consumption of alcohol. DESIGN: The sample included 67 young adults, who performed a smear of the oral mucosa and answered a questionnaire about recent and lifetime consumption of alcohol and other drugs. The methods used were sensitive to show the association between alcohol consumption and other drugs with the damage to oral cavity cells. RESULTS: DNA fragmentation index, mitochondrial functionality and cell viability, showed a significant difference between alcohol users and nonusers. However, there was no distinction between these parameters and different frequency consumption. Alcohol consumption, economic class and use of illicit drugs were related to the cytological parameters evaluated. CONCLUSIONS: This result shows the existence of cell damages among the evaluated students and can direct future studies towards more in-depth investigations of the mechanisms involved.

3-[(4-chlorophenyl)selanyl]-1-methyl-1H-indole ameliorates long-lasting depression- and anxiogenic-like behaviors and cognitive impairment in post-septic mice: Involvement of neuroimmune and oxidative hallmarks.

Only in the last decade the long-term consequences of sepsis started to be studied and even less attention has been given to the treatment of psychological symptoms of sepsis survivors. It is estimated that 60% of sepsis survivors have psychological disturbances, including depression, anxiety, and cognitive impairment. Although the causative factors remain largely poorly understood, blood-brain barrier (BBB) disturbances, neuroinflammation, and oxidative stress have been investigated. Therefore, we sought to explore if the immunomodulatory and antioxidant selenocompound 3-[(4-chlorophenyl)selanyl]-1-methyl-1H-indole (CMI) would be able to ameliorate long-term behavioral and biochemical alterations in sepsis survivors male Swiss mice. CMI treatment (1 mg/kg, given orally for seven consecutive days) attenuated depression- and anxiogenic-like behaviors and cognitive impairment present one month after the induction of sepsis (lipopolysaccharide, 5 mg/kg intraperitoneally). Meantime, CMI treatment modulated the number of neutrophils and levels of reactive species in neutrophils, lymphocytes, and monocytes. In addition, peripheral markers of liver and kidneys dysfunction (AST, ALT, urea, and creatinine) were reduced after CMI treatment in post-septic mice. Notably, CMI treatment to non-septic mice did not alter AST, ALT, urea, and creatinine levels, indicating the absence of acute hepatotoxicity and nephrotoxicity following CMI treatment. Noteworthy, CMI ameliorated BBB dysfunction induced by sepsis, modulating the expression of inflammation-associated genes (NFκB, IL-1ß, TNF-α, IDO, COX-2, iNOS, and BDNF) and markers of oxidative stress (reactive species, nitric oxide, and lipid peroxidation levels) in the prefrontal cortices and hippocampi of mice. In conclusion, we unraveled crucial molecular pathways that are impaired in post-septic mice and we present CMI as a promising therapeutic candidate aimed to manage the long-lasting behavioral alterations of sepsis survivors to improve their quality of life.

Atrazine exposure in gestation and breastfeeding affects Calomys laucha sperm cells.

The present study evaluated the effects of low doses of atrazine administered during gestation and breastfeeding on sperm characteristics of the wild rodent Calomys laucha. Adult females were divided into groups of 10 and administered different doses of atrazine through gavage, during gestational or breastfeeding period. At 3 months of age, the F1 adult male progeny of these females was evaluated. We observed a drastic reduction in the total and progressive motility of male sperm cells at all doses and during both the exposure periods. Moreover, the plasma membrane integrity of adult male sperm cells decreased at all doses of atrazine administered during the breastfeeding, whereas the membrane fluidity of these cells increased at all tested doses. Atrazine led to a decrease in the sperm mitochondrial functionality at all doses and during both exposure periods. The damage to the sperm DNA was higher in males exposed to the highest dose (1.0 mg/kg) during the gestation period, and in animals exposed to the lowest dose of atrazine (0.1 mg/kg) during breastfeeding period. Furthermore, the highest dose (1.0 mg/kg) of atrazine reduced the sperm concentration. Furthermore, the reduced levels of reactive oxygen species (ROS) were observed at all evaluated doses in males exposed during the gestation period. These results suggest that the administration of low doses of atrazine at critical periods of development may permanently reduce the sperm quality in C. laucha.

Life-time exposure to waterborne copper IV: Sperm quality parameters are negatively affected in the killifish Poecilia vivipara.

In previous studies, we have shown that copper (Cu) is significantly accumulated in various tissues of killifish Poecilia vivipara following chronic exposure. Also, we showed that chronic metal exposure disrupted energy production and growth in this species. In the present study, we aimed to evaluate if chronic exposure to this metal could also affect reproductive parameters of P. vivipara males (sperm quality). In order to test that, newborn (<24 h-old) fish were exposed to two concentrations of waterborne Cu (5 and 9 µg/L) for 345 days. After exposure, fish were euthanized and the testes were collected for sperm analysis. We could observe that exposed animals had reduced sperm motility and period of motility. Also, the sperm of exposed fish had reduced plasma membrane integrity, mitochondrial functionality and DNA integrity when compared to sperm of control animals. It is suggested that the well-known association of Cu with elevated oxidative damage, endocrine disruption and energetic disturbance are involved with the observed outcomes. The results obtained in the present study show that chronic exposure to environmentally relevant concentrations of waterborne Cu caused reductions in all parameters used to evaluate sperm quality. Therefore, it is concluded that life-time exposure to this metal may disrupt fish reproduction and negatively affect the maintenance of its populations.