Arsenic-Induced Sex Hormone Disruption: An Insight into Male Infertility.

Arsenic (As) is one of the most potent natural as well as anthropogenic metalloid toxicants that have various implications in the everyday life of humans. It is found in several chemical forms such as inorganic salt, organic salt, and arsine (gaseous form). Although it is mostly released via natural causes, there are many ways through which humans come in contact with As. Drinking water contamination by As is one of the major health concerns in various parts of the world. Arsenic exposure has the ability to induce adverse health effects including reproductive problems. Globally, around 15% of the couples are affected with infertility, of which about 20-30% are attributed to the male factor. Arsenic affects the normal development and function of sperm cells, tissue organization of the gonads, and also the sex hormone parameters. Stress induction is one of the implications of As exposure. Excessive stress leads to the release of glucocorticoids, which impact the oxidative balance in the body leading to overproduction of reactive oxygen species (ROS). This may in turn result in oxidative stress (OS) ultimately interfering with normal sperm and hormonal parameters. This study deals with As-induced OS and its association with sex hormone disruption as well as its effect on sperm and semen quality.

Mutagens in raw ewe milk in Orava region, northern Slovakia: metals.

The aim of this work was to determine the concentrations of selected mutagenic elements (As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Se) in raw ewe milk from undisturbed environment in Orava region, northern Slovakia. There are possible sources of some of the analyzed metals which may be distributed from the metallurgical plants located in the Ostrava region, Czech Republic, and Katowice, Poland. In total, forty milk samples were analyzed in June and August using an inductively coupled plasma optical emission spectrometry. The differences in elements concentrations between the seasonal periods were not significant except of iron (p < 0.0001). The concentrations of most of the metals in ewe milk were low and under the permissible or recommended limits. However, arsenic and selenium concentrations were elevated and could pose a risk of the mutagenic effect, particularly in children. The frequency of element occurrence in June was as follows: Se > Fe > As > Cu > Mn > Ni > Co > Pb > Cr > Cd, and in August: Se > Fe = As > Cu > Mn > Pb > Co > Ni > Cr > Cd. The correlation analysis revealed very strong positive correlation between Cu:Pb (p < 0.05), very strong negative correlation between Fe:Se (p < 0.05). The strong correlations were also found between other elements. The present study showed that milk produced in the relatively undisturbed environment might contain various mutagenic elements. The relationships between the elements might result in the additive or synergistic effects of elements and increase the risk of their mutagenic effects even in low concentrations. Therefore, attention must be paid to the monitoring of metals in the areas where food sources destined especially for child nutrition are produced.

Dose-Dependent Impact of Bee Pollen Supplementation on Macroscopic and Microscopic Structure of Femoral Bone in Rats.

Bee pollen has been successfully used as a feed additive with beneficial impacts on productive, reproductive, and immune conditions of animals. However, its effect on bone structure and bone health remains controversial. Therefore, the purpose of our study was to examine the impact of bee pollen supplementation on macroscopic and microscopic structure of a femoral bone using rats as suitable animal models. Male rats (1 month-old) were assigned into three groups: control (C group) that was fed a standard diet without bee pollen and two bee pollen supplemented groups (P1 and P2 groups) that received an experimental diet including 0.5% and 0.75% of bee pollen, respectively, for 3 months. A number of unfavorable effects of 0.75% bee pollen administration on bone weight, cortical bone thickness, calcium content, alkaline phosphatase activity, sizes of primary osteons' vascular canals, Haversian canals and secondary osteons in the cortical bone have been recorded, whereas these bone parameters were significantly decreased in the P2 group versus the C group. On the contrary, the concentration of 0.5% did not affect any of bone features mentioned above. In conclusion, the impact of bee pollen supplementation on femoral bone structure of rats depends on the dose used.

Determination of Essential and Toxic Elements in Raw Sheep's Milk from Area of Slovakia with Environmental Burden.

Concentrations of selected essential (Ca, Mg, Zn, Se, Fe, Cu) and toxic elements (As, Cd, Hg, Pb, Ni) were analyzed in milk of sheep (300 ewes of Tsigai breed) depending on lactation stage in area of Slovakia with slightly disturbed environment. To determine the real state of environment in the selected area, the study was extended to include analyses of the soil and animal feed (green pastures). Analyses of elements in soil were performed using ICP-OES and elements in feed and milk using the AAS-HG, AES-ICP, AAS-ETA, AAS-AMA, and AAS-F. Analyses of soil samples and feed samples revealed the presence of toxic elements in this area, but their concentrations did not affect concentrations in milk and were below the LOQ (limit quantification). Significant decrease of Ca as well as Mg and Zn (P < 0.05) concentrations was found between early and middle stages of lactation and significant increase between the middle and late of lactation period. Concentration of Se and Fe in milk during the lactation significantly increased (P < 0.05) in the last stage of lactation. Concentration of Cu in milk of sheep was below LOQ during all stages of lactation. In the milk of sheep during the lactation stages, the significant positive correlations between concentrations of the following elements were observed: Ca-Mg, Fe-Se, Ca-Se, Ca-Fe, Mg-Se, Mg-Fe, Ca-Zn, and Mg-Zn. The low concentrations, below LOQ of selected toxic elements in milk of ewes, indicate the safety of this milk for direct consumption or for further food processing.

Concentrations of toxic metals and essential elements in raw cow milk from areas with potentially undisturbed and highly disturbed environment in Slovakia.

Dietary composition and husbandry practices largely determine the essential trace element status and the toxic metal exposure of livestock and consequently their concentrations in animal products. The main objective of this study was to determine the real contamination of selected areas of Slovakia compared with existing Environmental regionalization of the Slovak Republic (SR) created by the Ministry of the Environment SR. Another aim of this study was to determine the content of essential and toxic metals in feed and milk of dairy cows on selected farms in Slovakia referred to the suitability of the use of milk from these areas to other food processing. This article deals with the analysis of the content of selected elements in feed and milk of dairy cows in the area of Novot (Northern Slovakia; area with undisturbed environment) and Cecejovce (Eastern Slovakia; highly disturbed environment). Eleven elements have been analyzed (essential elements: calcium, zinc, magnesium, selenium, iron, copper; toxic elements: arsenic, cadmium, mercury, lead, nickel). Samples of feed and milk were collected five times during the spring season and five times during the autumn season in 2016. Analysis of samples was performed in Eurofins Bel/Novamann (Nové Zámky, Slovak Republic). Analyses were performed by atomic absorption spectroscopy and atomic emission spectrometry. The obtained values of the individual elements in the feed and milk were evaluated by Student's t test. Significantly higher contents of essential elements Ca, Zn, Fe, Mg, and Cu were found in the feed of dairy cows in Cecejovce (P < 0.001). Significantly higher (P < 0.001) As and Ni content in feed in Cecejovce in autumn season was also recorded. However, the content of these elements in feed did not affect their milk content. The content of Ca, Zn, and Mg in milk was significantly (P < 0.001) higher in Novot in both investigated seasons. Some essential elements and toxic elements in feed and milk were below the LOQ (limit of quantification). Reduced content of Ca and Zn in milk in both farms compared with optimal limits, decreased Ca and Mg content in feed in Novot, and a significant increase of Ca and Mg in feed in Cecejovce in autumn season were found. It can be concluded, due to the low content of toxic elements on these farms, that the use of milk of dairy cows from these areas for direct use or for dairy product processing is appropriate and poses no health risk to the consumers.

Sperm motility and morphology changes in rats exposed to cadmium and diazinon.

BACKGROUND: Humans are ubiquitously exposed to multiple environmental contaminants. Consequences of combined action on the reproductive system remain unknown. This study aimed to assess single and joint effects of cadmium and diazinon exposure on sperm quality parameters. METHODS: Male adult Wistar rats were randomized into 4 groups of ten animals each. Group A was used as a control, animals from group B were exposed to cadmium (30 mg/L), rats from group C were administered with diazinon (40 mg/L), and rats from group D were exposed simultaneously to cadmium (30 mg/L) and diazinon (40 mg/L) via drinking water for 90 days. Sperm morphology and motility were evaluated using a bright field microscope and a computer-assisted semen analysis. RESULTS: The percentage of motile spermatozoa and morphologically normal sperm was markedly reduced in rats from the group B. Rats from the C group showed an increase in velocity parameters, amplitude of lateral head displacement, decrease in beat-cross frequency, and an increase in abnormal sperm morphology. Simultaneous coexposure to cadmium and diazinon increased distance and velocity parameters, and amplitude of lateral head displacement. Reductions were observed in straightness, linearity, wobble, and beat-cross frequency. The decreased normal sperm morphology rates were related to defects of the sperm tail. CONCLUSIONS: Exposure to cadmium and diazinon at relatively low doses impairs sperm quality and can reduce male fertility. Cadmium and diazinon caused significant changes on sperm morphology with varying effects on motility patterns. These parameters were significantly higher in the group D as compared to the group C. The findings have important implications for reproductive risk assessment of combined exposures to multiple chemicals.

Effect of separate and combined exposure of selenium and diazinon on rat sperm motility by computer assisted semen analysis.

Effects of selenium (Se) and diazinon (DZN) on sperm motility parameters in rats were investigated. Male rats received a separate dose of Se (2mgkg-1 b.w., intraperitoneally, 5mgL-1, per os in drinking water), diazinon (20mgkg-1 b.w., intraperitoneally, 40mgL-1, per os in drinking water), and in combination (Se+DZN) with the same dosage as in the separate administration. 36h an intraperitoneal (i.p.) and after 90days of per oral (p.o.) exposure, thirteen parameters of sperm motility were evaluated using a Computer Assisted Sperm Analyzer (CASA). Almost all the evaluated sperm motility parameters significantly decreased in Se p.o. exposed groups. In the Se i.p. group decrease was noted only in beat cross frequency (BCF) and progressive motility. Significant decline in the sperm motility, progressive motility, BCF and increase in amplitude of lateral head displacement (ALH) were recorded after DZN i.p. administration. In DZN p.o. group, significant increase in ALH, velocity average path (VAP) and curvilinear velocity (VCL) but decrease in progressive motility and BCF was detected. Se+DZN i.p. administration caused a significant decrease in motility, progressive motility and BCF. Per oral administration of Se+DZN decreased all motility parameters except LIN, WOB and ALH. Sperm abnormalities increased in all experimental conditions. Se and DZN negatively affected sperm structure and function in separate doses or in combination. No protective effect of Se was observed.

Effects of subchronic exposure to cadmium and diazinon on testis and epididymis in rats.

The present study aimed to elucidate the structural changes in testis and epididymis of adult rats following subchronic peroral administration of cadmium at 30 mg/L, diazinon at 40 mg/L, cadmium at 30 mg/L, and diazinon at 40 mg/L, respectively. At the end of 90-day experiment, the samples of the testes and epididymis were assayed by qualitative and quantitative histological methods. The testis and epididymis weights increased following exposure to cadmium and simultaneous exposure to cadmium and diazinon. Testicular damage following cadmium and diazinon coexposure was significantly less expressive than in groups with individual administration of these compounds. Cadmium caused a significant thickening of seminiferous epithelium, cellular degeneration, and necrosis. Desquamation of immature germ cells resulted in a significant increase of intraepithelial spaces and reduced tubule volume in all experimental groups. Vascular dilation and congestion were detected in the interstitial tissue. The changes in epididymal histology in the group exposed to cadmium and group exposed simultaneously included a reduction of epithelium, necrotic epithelial cells, vasoconstriction, and interstitial edema together with mononuclear cell infiltration. Results did not indicate a synergistic or any additional effect from the simultaneous administration of both toxicants. Further research is needed to determine the significance and the mechanism of the adverse effects.

Cobalt-induced hormonal and intracellular alterations in rat ovarian fragments in vitro.

The objective of this in vitro study was to examine dose-dependent changes in the secretion activity (progesterone, 17ß-estradiol and insulin-like growth factor-I) of rat ovarian fragments after experimental cobalt (Co) administration including the apoptotic potential of Co on rat ovarian fragments by evaluating the expression of apoptotic markers Bax and caspase-3. Ovarian fragments were incubated with cobalt sulphate (CoSO4.7H2O) at the doses 90, 170, 330 and 500 µg.mL(-1) for 24 h and compared with control group without Co addition. Release of progesterone (P4) 17ß-estradiol and insulin-like growth factor-I (IGF-I) by ovarian fragments was assessed by RIA, expression of Bax and caspase-3 by SDS-PAGE and Western blotting. Observations show that P4 release by ovarian fragments was significantly (P < 0.05) inhibited after cobalt sulphate addition at higher doses 170-500 µg.mL(-1) used in the study in comparison to control. However, cobalt sulphate addition did not cause any significant change in the release of 17ß-estradiol by ovarian fragments at all the doses used in the study (90-500 µg.mL(-1)) in comparison to control. On the contrary, IGF-I release by ovarian fragments was significantly (P < 0.05) stimulated after cobalt sulphate addition at the lowest dose 90 µg.mL(-1) in comparison to control, while other doses did not cause any significant change. Also, addition of cobalt sulphate decreased the expression of both the apoptotic peptides Bax and caspase-3 at the higher doses 170, 330 and 500 µg.mL(-1), but not at the lowest dose 90 µg.mL(-1) used in the study. Obtained results suggest Co induced (1) inhibition in secretion of steroid hormone progesterone, (2) dose-dependent increase in the release of growth factor IGF-I, and (3) decrease in the expression of markers of apoptosis (Bax and caspase-3) of rat ovarian fragments.

Changes in compact bone microstructure of rats subchronically exposed to cadmium.

BACKGROUND: Chronic exposure to cadmium (Cd), even at low concentrations, has an adverse impact on the skeletal system. Histologically, primary and secondary osteons as basic structural elements of compact bone can also be affected by several toxicants leading to changes in bone vascularization and mechanical properties of the bone. The current study was designed to investigate the effect of subchronic peroral exposure to Cd on femoral bone structure including histomorphometry of the osteons in adult male rats. In our study, 20 one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group, young males received a drinking water containing 30 mg of CdCl2/L, for 90 days. Ten one-month-old males without Cd intoxication served as a control group. After 90 days of daily peroral exposure, body weight, femoral weight, femoral length, cortical bone thickness and histological structure of the femora were analysed. RESULTS: We found that subchronic peroral application of Cd had no significant effect on body weight, femoral length and cortical bone thickness in adult rats. On the other hand, femoral weight was significantly increased (P < 0.05) in Cd-intoxicated rats. These rats also displayed different microstructure in the middle part of the compact bone where vascular canals expanded into central area of substantia compacta and supplied primary and secondary osteons. Additionally, a few resorption lacunae which are connected with an early stage of osteoporosis were identified in these individuals. Histomorphometrical evaluations showed that all variables (area, perimeter, maximum and minimum diameter) of the primary osteons' vascular canals, Haversian canals and secondary osteons were significantly decreased (P < 0.05) in the Cd group rats. This fact points to alterations in bone vascularization. CONCLUSIONS: Subchronic peroral exposure to Cd significantly influences femoral weight and histological structure of compact bone in adult male rats. It induces an early stage of osteoporosis and causes reduced bone vascularization. Histomorphometrical changes of primary and secondary osteons allow for the conclusion that the bone mechanical properties could be weakened in the Cd group rats. The current study significantly expands the knowledge on damaging action of Cd on the bone.

Simultaneous subchronic exposure to selenium and diazinon as possible risk factor for osteoporosis in adult male rats.

BACKGROUND: Osteoporosis and its main health outcome, fragility fractures, are large and escalating health problems. Skeletal damage may be the critical result of low-level prolonged exposure to several xenobiotics in the general population, but the mechanisms of their adverse effects are not clearly understood. The current study was aimed to investigate the possible ability of simultaneous subchronic peroral administration of selenium (Se) and diazinon (DZN) to induce changes in bone of adult male rats.In our study, twenty 1-month-old male Wistar rats were randomly divided into two experimental groups. In the first group, young males were exposed to 5 mg Na2SeO3/L and 40 mg of DZN/L in drinking water, for 90 days. Ten 1-month-old males without Se and DZN intoxication served as a control group. At the end of the experiment, macroscopic and microscopic structures of the femurs were analysed using analytical scales, sliding instrument, and polarized light microscopy. RESULTS: The body weight, femoral length and cortical bone thickness were significantly decreased in rats simultaneously exposed to Se and DZN (P < 0.05). These rats also displayed different microstructure in the middle part of the compact bone where vascular canals expanded into central area of substantia compacta. The canals occurred only near endosteal surfaces in rats from the control group. Additionally, a smaller number of primary and secondary osteons, as well as a few resorption lacunae were observed near endosteal surfaces in rats simultaneously administered to Se and DZN. The resorption lacunae as typical structures of bone resorption manifestation are connected with an early stage of osteoporosis. Histomorphometric analysis revealed that area, perimeter, maximum and minimum diameters of primary osteons' vascular canals were significantly increased (P < 0.05) in the Se-DZN-exposed rats. On the other hand, all measured variables of Haversian canals and secondary osteons were considerable reduced (P < 0.05) in these rats. CONCLUSIONS: Simultaneous subchronic peroral exposure to Se and DZN induces changes in macroscopic and microscopic structures of the femurs in adult male rats, and also it can be considered as possible risk factor for osteoporosis. The current study contributes to the knowledge on damaging impact of several xenobiotics on the bone.

Structural changes in femoral bone tissue of rats after subchronic peroral exposure to selenium.

BACKGROUND: The role of selenium (Se) on bone microarchitecture is still poorly understood. The present study aims to investigate the macroscopic and microscopic structures of femoral bone tissue in adult male rats after subchronic peroral administration of Se. METHODS: Twenty one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group (Se group) young males were exposed to 5 mg Na(2)SeO(3)/L in drinking water, for 90 days. Ten one-month-old males without Se administration served as a control group. At the end of the experiment, macroscopic and microscopic structures of the femurs were analysed using analytical scales, sliding instrument, and polarized light microscopy. RESULTS: The body weight, femoral length and cortical bone thickness were significantly decreased in Se group rats. These rats also displayed different microstructure in the middle part of the femur, both in medial and lateral views, where vascular canals expanded into the central area of the bone while, in control rats, these canals occurred only near the endosteal surfaces. Additionally, a smaller number of primary and secondary osteons was identified in Se group rats. Histomorphometric analyses revealed significant increases for area, perimeter, maximum and minimum diameters of primary osteons' vascular canals but significant reductions for all measured variables of Haversian canals and secondary osteons. CONCLUSIONS: Se negatively affected the macroscopic and microscopic structures of femoral bone tissue in adult male rats. The results contribute to the knowledge on damaging impact of Se on bone.

Quantitative histological analysis of the mouse testis after the long-term administration of nickel in feed.

In this study, the effects of nickel chloride (NiCl(2)) applied per os on testis histopathology and morphometry of mice were investigated. The metal was applied in pellets at a dose of 10 mg NiCl(2)/kg bw to male mice 4 weeks of age. After 3, 6, 9 and 12 weeks of nickel administration, the relative volume of whole seminiferous tubule, germinal epithelium, tubule lumen, interstitium and blood vessels as well as the diameter of seminiferous tubules were determined in the experimental and corresponding control groups. Microscopic examination of testis showed significant changes in all nickel-exposed groups. The degeneration of germinal epithelium, with released germ cells into the lumen of the tubules, and occurrence of empty spaces in the seminiferous epithelium were found in all experimental groups. The changes in the testes were time-dependent. The relative volume of empty spaces in the seminiferous epithelium significantly increased (P < 0.001) in all experimental groups when compared with the corresponding control. A significant decrease in the relative volume of seminiferous epithelium was observed after 6 and 12 weeks of Ni-exposure. The increased luminization of the tubules was found after 6 (P < 0.001), 9 (P < 0.01) and 12 (P < 0.001) weeks. Interstitial tissue significantly decreased after 6 and 9 weeks of Ni exposure and increased after 12 weeks of Ni intake. The seminiferous tubule diameter significantly (P < 0.001) decreased after 12 weeks. Results of this study report a serious, time-dependent changes in the testes, mainly in the germinal epithelium, after a peroral intake of nickel.

In vivo and in vitro effect of bendiocarb on rabbit testicular structure and spermatozoa motility.

In this study the effect of bendiocarb on the rabbit testicular structure and spermatozoa motility was investigated. For testicular structure evaluation the animals were fed with bendiocarb tablets daily at a dose of 5 mg/kg of body weight for 13 days. The relative volume of the germinal epithelium, interstitium and lumen was measured. The testicular structure evaluation showed decreased relative volume of germinal epithelium in both experimental groups in comparison with the control group. The relative volume of the interstitium was increased in both experimental groups. An increase of the relative volume of the lumen was registered also in both experimental groups. Qualitative analysis detected a dilatation of blood vessels in the interstitium, undulation of the basal membrane and some empty spaces in the germinal epithelium after bendiocarb administration. The spermatozoa motility was evaluated by the computer assisted semen analyzer (CASA) method in various time intervals (0-180 minutes) and the bendiocarb concentration in the culture medium added to experimental groups varied from 0.054 to 0.268 mg/mL. Spermatozoa motility and progressive motility significantly decreased with increased bendiocarb administration and with extending the period of incubation. For other fine motility parameters, a decrease dependent on the time of incubation and on the bendiocarb concentration almost in all experimental groups in comparison to the control was detected. These results clearly suggest that in vitro also in vivo bendiocarb administration decrease male fertility.

Effects of a single intraperitoneal administration of cadmium on femoral bone structure in male rats.

BACKGROUND: Exposure to cadmium (Cd) is considered a risk factor for various bone diseases in humans and experimental animals. This study investigated the acute effects of Cd on femoral bone structure of adult male rats after a single intraperitoneal administration. METHODS: Ten 4-month-old male Wistar rats were injected intraperitoneally with a single dose of 2 mg CdCl2/kg body weight and killed 36 h after the Cd had been injected. Ten 4-month-old males served as a control group. Differences in body weight, femoral weight, femoral length and histological structure of the femur were evaluated between the two groups of rats. The unpaired Student's t-test was used for establishment of statistical significance. RESULTS: A single intraperitoneal administration of Cd had no significant effect on the body weight, femoral weight or femoral length. On the other hand, histological changes were significant. Rats exposed to Cd had significantly higher values of area, perimeter, maximum and minimum diameters of the primary osteons' vascular canals and Haversian canals. In contrast, a significant decrease in all variables of the secondary osteons was observed in these rats. CONCLUSIONS: The results indicate that, as expected, a single intraperitoneal administration of 2 mg CdCl2/kg body weight had no impact on macroscopic structure of rat's femora; however, it affected the size of vascular canals of primary osteons, Haversian canals, and secondary osteons.

Nickel induced structural and functional alterations in mouse Leydig cells in vitro.

The present study was aimed at investigating effects of nickel (NiCl(2)) on secretion of testosterone (T), cell viability, ultrastructure and apoptosis in mouse Leydig cells. Testosterone release was measured after 48h of culture with 15.67, 31.25, 62.5, 125, 250, 500 and 1000µmol/L NiCl(2) or without NiCl(2) using radioimmunoassay. Cell viability was assessed by a MTT (metabolic activity assay). Quantification of apoptotic cells was performed using TUNEL assay and the ultrastructural changes were analyzed using transmission electron microscopy. The viability was decreased after addition of ≥250µmol/L NiCl(2). A concentration-dependent depression of T production was observed. The percentage of apoptotic cells was significantly increased only after addition of 125, 250 and 1000µmol/L NiCl(2). After addition of ≥250µmol/L NiCl(2) higher incidence of euchromatin was observed. Lipid droplets and vacuoles in cytoplasm were increased after addition of ≥125µmol/L NiCl(2). NiCl(2) induced decrease in numbers of mitochondria and smooth endoplasmic reticulum after treatment with ≥500µmol/L NiCl(2). Our findings suggest a negative effect of NiCl(2) on steroidogenesis, viability, apoptosis and ultrastructure of mouse Leydig cells.

Cadmium toxicity at low concentration on rabbit spermatozoa motility, morphology and membrane integrity in vitro.

In this study the effect of cadmium on various parameters of spermatozoa motility, morphology as well as on the spermatozoa membrane integrity in rabbits was analyzed in vitro, experimental concentrations ranging from 0.62 to 0.98 micro g CdCl(2)/mL. Pooled rabbit (n = 5) semen was cultured in vitro with cadmium and subsequently diluted to various experimental concentrations apart from control which received no cadmium exposure. Using computer assisted semen analysis method (CASA) we detected decrease of total motility with in the higher concentration range at Time 0. However, with increasing time (after 1 and 2 h of culture), cadmium exerted deleterious effect leading to significant motility reduction in comparison to control. A similar trend was exhibited in case of progressive motility, too. Most of the spermatozoa distance and velocity parameters detected no significant change in comparison to control at the beginning of culture (Time 0), although the toxic effect became significant (P < 0.05) with the passage of culture time (Times 1 and 2 h) in all concentrations. Analysis of spermatozoa morphology detected significant (P < 0.05) alterations at higher concentrations. At higher concentrations acrosomal changes, head without flagellum/separated flagellum, broken flagellum and other abnormalities were significantly higher (P < 0.05), while knob-twisted flagellum and small heads differed significantly (P < 0.05) in comparison to control at all concentrations. In regards to flagellum torso, flagellum ball and retention of cytoplasmic drop statistically higher values (P < 0.05) were noted at the maxium experimental concentration only. Annexin analysis for detection of spermatozoa with disordered membranes revealed higher occurrence of positive spermatozoa in cadmium exposed groups. Annexin-positive reactions suggested alterations in anterior part of head (acrosome) and in flagellum (mitochondrial segment) of spermatozoa. This paper underlines that cadmium is highly toxic for rabbit spermatozoa, as visualized by the toxic effects on parameters of spermatozoa motility, morphology and membrane integrity. The toxic effect is more drastic at higher concentrations. This study also indicates that cadmium requires a minimum one hour incubation time to exert its deletorious effects on various parameters of spermatozoa, particularly at low concentrations.

Mercury-induced alterations in rat kidneys and testes in vivo.

In this study effects of mercury administration on the kidney and testicular structure of adult rats were evaluated. Rats received mercury (HgCl2) in single intraperitoneal dose 20 mg HgCl2 (group A), 10 mg HgCl2 (group B) and 5 mg HgCl2 (group C) per kilogram of body weight and were killed after 48 hours following mercury administration. After the preparation of histological samples the results were compared with control group (K). In kidney decreased diameters of glomeruli and renal corpuscles, damaged tubules with affected quality of tubular cells and infiltration of interstitium were detected. Quantitative analysis demonstrated increased relative volume of tubules and renal corpuscles. Also the number of nuclei and glomeruli was increased in all experimental groups. The diameter of glomeruli and renal corpuscles was decreased. In testis undulation of basal membrane, dilatation of blood vessels in interstitium and occurrence of empty spaces in germinal epithelium were observed. Decreased relative volume of germinal epithelium, increased relative volume of interstitium and increased apoptosis occurrence suggest damaged interstitium and revealed occurrence of edemas. The relative volume of seminiferous tubules showed higher luminization. The number of nuclei was decreased in all experimental groups what is in positive relation with occurrence of empty spaces. Also other evaluated criteria demonstrated significant differences between control group and experimental groups. This study reports a negative effect of mercury on the structure and function of kidney and testes.

Lead-induced alterations in rat kidneys and testes in vivo.

The purpose of this study was to assess the effects of lead administration on the kidney and testicular structure of adult rats. Rats received lead (PbNO(3)) in single intraperitoneal dose 50 mg/kg (group A), 25 mg/kg (group B) and 12.5 mg (group C) per kilogram of body weight and were killed 48 h following lead administration. After the preparation of histological samples the results were compared with control. After the lead administration dilated Bowman's capsules and blood vessels in interstitium of kidney with evident hemorrhagic alterations were noted. Quantitative analysis determined increased relative volume of interstitium and tubules. Also, the diameter of renal corpuscules, diameter of glomeruli and diameter of Bowman's capsule were significantly increased, especially in group A, with the highest lead concentration. In testes, dilatation of blood capillaries in interstitium, undulation of basal membrane and occurrence of empty spaces in seminiferous epithelium were detected. An apoptosis assay confirmed increased incidence of apoptosis in the spermatogenetic cells after the lead administration. Also further morphometric analysis showed significant differences in evaluated parameters between control and treated groups. The number of cell nuclei was decreased in lead-treated groups, which is concerned with the occurrence of empty spaces as well as with the higher apoptosis incidence in germinal epithelium. This study reports a negative effect of lead on the structure and function of kidney and testes.

Cobalt-induced alterations in hamster testes in vivo.

The purpose of this study was to assess the effects of cobalt on the testicular structure of adult golden hamsters. Hamsters in group A received cobalt (CoCl2) in single intraperitoneal dose 20 mg/kg, in group B 10 mg/kg and in group C 5 mg CoCl2/kg body weight and were killed fourty eight hours after cobalt administration. Afer a preparation of histological samples the results were compared with control. After a cobalt administration dilatation of blood capillaries in interstitium, undulation of basal membrane and occurrence of empty spaces in seminiferous epithelium was detected. Morphometric analysis showed that in all cobalt-treated groups the relative volume of seminiferous epithelium was significantly decreased. In the relative volume of interstitium a significant increase was found between control group and experimental groups. After cobalt administration we have found linear non-significant decrease. Evaluation of diameter seminiferous tubules found increase of this parameter in the all experimental group in comparison with control. Height of seminiferous epithelium was relatively constant and in all groups but the difference between control and group A was significant (P < 0.05). Analysis of the lumen diameter of seminiferous tubules detected significantly increase mainly group B. Evaluation of the number of cell nuclei per a constant area detected an increase of this parameter in experimental group. Results of this study report a negative effect of cobalt on structure and function of testes.