Robert Toman

Concentration of Copper, Iron, Zinc, Cadmium, Lead, and Nickel in Bull and Ram Semen and Relation to the Occurrence of Pathological Spermatozoa

Abstract In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (p < 0.0001) in ram semen in comparison with bull semen. The zinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79 ± 4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17 ± 3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r = 0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r = 0.98), nickel and separated tail (r = 0.77), separated tail and total number of pathological spermatozoa (r = 0.69), knob twisted tail and retention of cytoplasmic drop (r = 0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r = 0.71), copper and separated tail (r = 0.70), and between iron and tail torso (r = 0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality.

Accumulation of Lead, Cadmium, and Mercury in Liver and Kidney of the Brown Hare (Lepus europaeus) in Relation to the Season, Age, and Sex in the West Slovakian Lowland

Abstract Concentrations of lead, cadmium, and mercury in liver and kidneys of brown hares in relation to season, age, and sex were investigated. Over a period of one year 74 hares, 36 males and 38 females were analyzed. The concentrations of lead and cadmium were analyzed by AAS and mercury was determined by mercury vapor technique. The median concentration of lead in liver as well as in kidney in relation to the season is the highest in winter period in comparison with spring, summer, and summer period (p<0.001). The highest concentration of cadmium was found in winter, but the differences were not significant. In mercury, we report significantly higher (p<0.001) median concentrations in liver (0.023 mg kg−1) as well as in kidney (0.068 mg kg−1) in winter period in comparison with all other observed periods. In relation to age the concentrations in lead and mercury in liver and kidney were very similar, without significant differences. In cadmium we report significantly higher concentrations in the liver as well as kidney in adult animals (0.154 and 1.521 mg kg−1) in comparison with juvenile animals (0.048 and 0.582 mg kg−1, respectively). In comparison of the female and male brown hares we found significantly higher (p<0.05) median concentration of lead in the liver of males (0.216 mg kg−1) than in females (0.127 mg kg−1) and the level of cadmium is significantly higher (p<0.001) in females (1.464 mg kg−1) than in male brown hares (1.384 mg kg−1).

Distribution of cadmium and lead in liver and kidney of some wild animals in Slovakia

Abstract The content of cadmium and lead, as risk factors of environment, in liver and kidneys of wild animals as brown hare (Lepus europaeus), yellow-necked mouse (Apodemus flavicollis), wood mouse (Cleithrionomys glareolus), and red deer (Cervus elaphus) were studied. Samples were analyzed by the atomic absorption spectrophotometry method (AAS). The highest levels of cadmium were found in kidneys (0.213–2.387 mg/kg) of all animal species. The concentration of cadmium in liver was 0.032–0.258 mg/kg. The analysis of lead showed that the concentration of this element was higher in kidneys of yellow-necked mouse and wood mouse (0.503–0.780 mg/kg) than in liver (0.177–0.268 mg/kg). In brown hare and red deer a higher accumulation of lead in liver (0.221–1.904 mg/kg) in comparison with kidneys (0.115–0.561 mg/kg) is reported.

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.

Concentration of Copper, Iron, Zinc, Cadmium, Lead, and Nickel in Boar Semen and Relation to the Spermatozoa Quality

Abstract The concentration of copper, iron, zinc, cadmium, lead, and nickel as well as its relation to spermatozoa quality was investigated. The semen samples were analyzed by atomic absorption spectrophotometry (AAS). The concentration of copper in boar semen was 1.64 ± 0.28 mg kg−1 and of iron 16.14 ± 10.35 mg kg−1. The concentration of zinc in boar semen reached an average value of 171.74 ± 64.72 mg kg−1 and the level of cadmium reached 0.01–0.16 mg kg−1 with the average value of 0.05 mg kg−1. The analysis of lead showed that the concentration of this element in boar semen was 0.02 ± 0.03 mg kg−1 and the average level of nickel was 0.06 ± 0.08 mg kg−1. The total percentage of pathological spermatozoa was 9.82 ± 1.47%. Detail analysis determined 3.18% of separated flagellum, 2.26% knob twisted flagellum, 0.88% flagellum torso, 0.85% flagellum ball, 0.42% broken flagellum, 0.23% retention of the cytoplasmic drop, 0.14% small heads, 0.03% large heads, and 1.83% forms other of pathological changes. Correlation analysis showed significant (p<0.05) positive correlation between copper and lead (r = 0.52). High correlation between small head and knob twisted tail (r = 0.67), small head and broken flagellum (r = 0.88) as well as between small head and total number of pathological spermatozoa (r = 0.73) was determined.

Seminal Concentration of Trace Elements in Fox and Relationships to Spermatozoa Quality

Concentrations of copper, zinc, iron, cadmium, lead, and nickel in the semen of foxes (Vulpes vulpes, n = 10), microscopic analysis of occurrence of pathological spermatozoa, and correlations of these elements with pathological forms were studied. Samples were analyzed by using an atomic absorption spectrophotometer. For analysis of pathological spermatozoa semen samples fixed with Hancocks solution and stained with Giemsa were prepared. For each fox at least 1000 spermatozoa were evaluated. The concentrations of copper, zinc, and iron in semen of foxes were found to be 2.16 ± 0.53 mg/kg, 13.09 ± 5.22 mg/kg, and 33.16 ± 24.36 mg/kg, respectively, on wet weight basis. Concentration of cadmium was low (0.07 ± 0.05 mg/kg). The levels of lead and nickel in the semen of foxes were 0.08 ± 0.06 mg/kg and 0.35 ± 0.24 mg/kg, respectively. The total percentage of pathological spermatozoa was 7.76 ± 1.33% with predominancy of knob twisted flagellum, separated flagellum, and broken flagellum. In relation to trace elements the analysis showed significant (p < 0.05) correlation between copper and lead (r = −0.85), copper and other forms of pathological spermatozoa (r = −0.72), zinc and broken flagellum (r = −0.69), iron and retention of cytoplasmic drop (r = 0.87), cadmium and separated flagellum (r = −0.68), and between cadmium retention of cytoplasmic drop (r = 0.87). This study detected significant effects of various trace elements in normal fox semen on the spermatozoa quality.

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 μ g CdCl2/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.

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.

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.

Consumption of bee pollen affects rat ovarian functions

The aim of this study was to examine possible effects of bee pollen added to the feed mixture (FM) on rat ovarian functions (secretion activity and apoptosis). We evaluated the bee pollen effect on the release of insulin-like growth factor I (IGF-I) and steroid hormones (progesterone and estradiol), as well as on the expression of markers of apoptosis (Bcl-2, Bax and caspase-3) in rat ovarian fragments. Female rats (n = 15) were fed during 90 days by FM without or with rape seed bee pollen in dose either 3 kg/1000 kg FM or 5 kg/1000 kg FM. Fragments of ovaries isolated from rats of each group (totally 72 pieces) were incubated for 24 h. Hormonal secretion into the culture medium was detected by RIA. The markers of apoptosis were evaluated by Western blotting. It was observed that IGF-I release by rat ovarian fragments was significantly (p < 0.05) decreased; on the other hand, progesterone and estradiol secretion was increased after bee pollen treatment at dose 5 kg/1000 kg FM but not at 3 kg/1000 FM. Accumulation of Bcl-2 was increased by bee pollen added at 3 kg/1000 kg FM, but not at higher dose. Accumulation of Bax was increased in ovaries of rats fed by bee pollen at doses either 3 or 5 kg/1000 kg FM, whilst accumulation of caspase-3 increased after feeding with bee pollen at dose 5 kg/1000 kg FM, but not at 3 kg/1000 kg FM. Our results contribute to new insights regarding the effect of bee pollen on both secretion activity (release of growth factor IGF-I and steroid hormones progesterone and estradiol) and apoptosis (anti- and pro-apoptotic markers Bcl-2, Bax and caspase-3). Bee pollen is shown to be a potent regulator of rat ovarian functions.