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.

High-speed centrifugation of extender of freeze-thaw boar semen.

In semen cryopreservation, egg yolk is still widely used as a non-penetrating cryoprotectant. Much has been developed in the search for alternatives for this biological product. This work aimed to evaluate the processed egg yolk through ultracentrifugation and/or sonication in the cryopreservation of swine semen. Twenty-seven semen doses were purchased from a commercial boar stud and processed for cryopreservation using egg yolk lactose 11% (control) extender, processed using two different methods: high-speed centrifugation and sonication. Then, they were submitted to freeze-thawing protocol and were assessed for kinematic and cell structural parameters. Samples in which extenders underwent centrifugation had better results in velocity parameters, meanwhile those that only sonication was performed had poorest results in this parameter. The preservation of the membrane and mitochondria structure had better results when the diluent was only centrifuged in comparison with the other treatments. Therefore, centrifugation of extender containing egg yolk is important for better cryopreservation of swine semen.

Use of trehalose in the semen cryopreservation of Amazonian catfish Leiarius marmoratus.

The current study assessed a semen cryopreservation protocol in the Amazonian catfish Leiarius marmoratus, a freshwater fish, of rheophilic behavior, and of great importance for Brazilian fish farming. Eight males (n = 8) were stripped and the semen was cryopreserved if total motility in fresh semen was higher than 80%. The external cryoprotectant Trehalose was then diluted in Beltsvile Thawing Solution (BTS) extender in the following concentrations: 50, 100, 150, and 200 mM. Semen samples were diluted in the media (1:9 v/v) being tested, then frozen in a container with nitrogen vapor (dryshipper), and stored in liquid nitrogen at -196 °C. Motility parameters assessed post-thawing were performed by CASA-system and sperm cell integrity analyses (membrane integrity, DNA integrity, and mitochondrial function) were performed through fluorescence microscopy. As a result, no significant statistical difference was observed between treatments, independently of Trehalose concentrations tested in the following post-thawing analysis: membrane integrity, DNA integrity, mitochondrial functionality, and sperm motility duration. As of total and progressive motilities, the treatment containing 50 mM trehalose (15.6 and 9.5%, respectively), exhibited inferior results when compared to treatments with 150 mM (22.9 and 17.7%, respectively) and 200 mM (31.4 and 26.3%, respectively) trehalose concentrations (P < 0.05); however, it did not differ from the treatment with 100 mM trehalose (18.6 and 15.3%, respectively). Therefore, treatments with trehalose at higher concentrations exhibited superior results when compared to other treatments in in vitro motility parameters for L. marmoratus.

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 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.

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.

Occupational exposure to 50 ppm isoflurane impairs sperm parameters in mice.

OBJECTIVE: To assess the effect of occupational exposure to isoflurane on the sperm quality of mice, and to investigate whether cessation of exposure can restore seminal quality. METHODS: Mice were exposed to 50 ppm of isoflurane for 5 hours per day over a 30-day period. Following this exposure, they were euthanized at predetermined post-exposure intervals, and their sperm samples were analyzed for kinetics and viability parameters. RESULTS: Occupational exposure to isoflurane can adversely affect sperm at 50 ppm, a concentration deemed safe by international occupational health conventions in some countries. These effects are marked by oxidative stress, mitochondrial dysfunction, and genomic damage, alongside alterations in sperm kinetics and acrosomal integrity. Furthermore, cessation of exposure doesn't guarantee restoration of cellular quality. CONCLUSIONS: Isoflurane at 50 ppm, previously deemed safe by some countries, impairs sperm kinetics and viability, potentially directing the spermatogenic process towards infertility and apoptosis.

Capillary electroporation affects the expression of miRNA-122-5p from bull sperm cells.

Sperm-mediated gene transfer (SMGT) has a potential application in the generation of transgenic animals. Capillary electroporation consists of the application of electrical pulses, resulting in an increased transfection rate. Little is known about the impacts of the transfection of exogenous DNA on sperm epigenetics. MicroRNAs are epigenetic factors that are related to sperm motility. MiRNA-122-5p regulates genes that influence motility, and consequently, the fertilizing potential of sperm. Therefore, we aimed at identifying whether epigenetic factors such as microRNAs could be altered after DNA transfection, using the capillary electroporation technique. In this study, bull sperm was electroporated using voltages of 600 V, 1500 V, and 0 V (control group), with or without exogenous DNA. Parameters of sperm quality were analyzed using CASA and flow cytometry, and expression of the miRNA-122-5p was analyzed using RT-qPCR. It was observed that electroporation increased the internalization of exogenous DNA (P < 0.05), but did not impair the mitochondrial activity (P > 0.05). It reduced sperm motility (P < 0.05). The expression of miRNA-122-5p was upregulated in sperm electroporated at 1500 V, and the presence of exogenous DNA did not affect its expression. Thus, we can conclude that electroporation influences the expression of miRNA-122-5p from bull sperm cells.

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.

Effects of exposure to triphenyltin (TPT) contaminant on sperm activity in adulthood of Calomys laucha exposed through breastfeeding.

Triphenyltin (TPT) is an organotin compound (OT), primarily used in agriculture and in the composition of antifouling paints for ships worldwide. Studies have showed its effects as an endocrine disrupter in several organisms by preventing enzymatic expression and causing reproductive toxicity. This study aimed to evaluate the effects of exposure to TPT, via breastfeeding, on reproductive physiology in the Calomys laucha species. The experimental design was compound of five groups, two controls and three with different doses of TPT. Moreover, females were exposed by gavage to the TPT for 20 days, from the 1st day postpartum to the 21st postnatal day (PND). Then, the pups were euthanized and the kinetics, organelles, and biochemistry of the sperm were evaluated. The results presented a reduction in total motility in the groups exposed to TPT. Regarding cellular organelles analysis, a loss in membrane integrity was evidenced; the functionality of mitochondria showed diminution followed by increased acrosome reaction. In conclusion, the TPT causes alteration of the reproductive parameters, decreasing the activity and sperm quality in individuals exposed in the breastfeeding phase.

Dimethylsulfoxide, methanol and methylglycol in the seminal cryopreservation of Suruvi, Steindachneridion scriptum.

The aim of this study was to analyze the effect of 5%, 7.5%, 10%, 12.5% and 15% dimethylsulfoxide (DMSO), methanol and methylglycol alcohols on the cryopreservation of sperm from Steindachneridion scriptum. Male specimens (n = 15) were obtained from Pisciculture and sperm samples were collected by abdominal massage. Post collection the fresh sperm sample was diluted in the Beltsville Thawing Solution and sperm motility was evaluated. Results indicated that the most precise parameters for total and progressive motility were obtained with the use of methylglycol (all concentrations) and 7.5% and 10% methanol (P < 0.05). The motility of the sperm was sustained for the longest time period when 5%, 7.5% and 15% DMSO was used; similar results were also seen for 5% methanol and methylglycol at 7.5%, 10%, 12.5%, and 15% concentration (P < 0.05). With respect to reactive oxygen species it was observed that the production of ROS decreased only in presence of 5% methylglycol but not when DMSO (5%) was used (P < 0.05). Although the use of methanol (12.5%) allowed for a lesser membrane fluidity as compared to DMSO 12.5% (P < 0.05), membrane functional integrity was greater with 10% and 12.5% DMSO (P < 0.05) as compared to 10% methanol or 5% methylglycol (P > 0.05). Additionally, when major mitochondrial functionalities were assessed it was observed that the values obtained with use of 12.5% and 15% DMSO were comparable to all except 5% methyglycol (P < 0.05). In conclusion, the results of the present study indicate that 7.5% methylglycol was the most effective treatment for the cryopreservation of the S. scriptum sperm.

Immunolocalization of Leptin and its Receptor (ObR-b) in Equine Placenta at Term and Plasma Level Measurement in the Late Gestation.

Placental tissues from humans, rodents, and farm animal contain leptin and its receptor. Expression of leptin has already been described in horses, although there is no description about immunolocalization of leptin and its receptor. The aim of the present study was to investigate the presence and distribution of leptin and ObR-b in the equine placenta at term by immunofluorescence staining, and the changes on plasma leptin concentrations during late gestation. The present study involved eight Criollo-type mares carrying healthy pregnancies. Blood samples were collected during the third trimester of gestation, at foaling, and at 24 hours after foaling. Leptin concentrations were analyzed via radioimmunoassay. Plasma leptin concentrations did not change from the 8th to the 10th month of gestation and displayed a discrete decrease 24 hours after parturition (P = .07). The expression of leptin and ObR-b was observed in the cytoplasm of pseudostratified epithelial cells in the areolar region and in the epithelium of microcotyledons. Also, leptin receptor was allocated in the apical surface of the cells. The presence of leptin and its receptor (ObR-b) in the placenta of mares at term supports an endocrine and autocrine/paracrine action of leptin within this organ.

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.

Low atrazine dosages reduce sperm quality of Calomys laucha mice.

Reproductive effects caused by the exposure to environmentally relevant dosages of atrazine on wild animals are poorly understood. This study evaluated the effects of three dosages of atrazine on sperm parameters of adult Calomys laucha males. Adult mice were orally exposed to dosages of 0 (water and vehicle control), 0.1, 1, and 10 mg/kg of animal weight for a 21-day period. Following exposure, analyses were performed to determine sperm motility parameters, plasma membrane integrity and fluidity, mitochondrial functionality, acrosome integrity, DNA damage, lipid peroxidation, and production of reactive oxygen species (ROS) in the sperm samples. Total and progressive motility were reduced in all dosages in comparison to control groups. Membrane integrity and mitochondrial functionality of sperm were reduced in all dosages, and the sperm membrane fluidity increased in the higher dosages of atrazine (1 and 10 mg/kg), in comparison with the vehicle control. A decrease in the acrosome integrity was noted at 10 mg/kg of atrazine, compared to the control groups. The integrity of DNA, ROS generation, and lipid peroxidation of sperm showed no significant differences when compared with the control groups. These results suggest that exposure to low dosages of atrazine can affect sperm parameters of Calomys laucha and therefore reduce the reproductive capacity of wild rodent species.

The herbicide atrazine affects sperm quality and the expression of antioxidant and spermatogenesis genes in zebrafish testes.

The herbicide atrazine (ATZ) is used worldwide in the control of annual grasses and broad-leaved weeds. The present study evaluated sperm quality parameters in zebrafish Danio rerio after 11-day exposure to nominal ATZ concentrations of 2, 10, and 100 µg L-1. All ATZ concentrations caused a decrease in motility, mitochondrial functionality, and membrane integrity, as measured using conventional microscopy or fluorescence microscopy with specific probes. The DNA integrity of sperm was not affected. The levels of expression of genes related to spermatogenesis, antioxidant defenses, and DNA repair were also investigated using RT-qPCR. The ATZ caused transcriptional repression of the spermatogenesis-related genes SRD5A2 and CFTR, the antioxidant defense genes SOD2 and GPX4B, and the DNA repair gene XPC. This is the first study to show that environmentally relevant concentrations of ATZ significantly affect the sperm quality in fish, possibly resulting in reduced fertility rates. In addition, we showed that the repression of genes related to spermatogenesis and cellular defense could be part of the mechanisms involved in the ATZ toxicity in the testes of male fish.