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.     

Study of the genetic variability of porcine circovirus type 2 detected in Serbia and Slovenia

Recent variants of porcine circovirus type 2 (PCV2) were obtained from tissues of domestic pigs with porcine circovirus associated disease and from randomly selected wild boar samples from Serbia and Slovenia. A 450-base-pair nucleotide sequence was obtained by PCR from the ORF2. The derived nucleotide and amino acid sequences were aligned and compared to the corresponding region of closely related PCV2 sequences determined in previous years and retrieved from the GenBank. The 30 Serbian and 17 Slovenian PCV2 sequences clustered into three previously determined genotypes (PCV2a: 7), (PCV2b: 38) and (PCV2d: 2). Three major variable regions, concerning 29 amino acid position substitutions within the ORF2, were observed, which further supports the segregation of the detected strains into three separate genotypes. This study indicates that PCV2b is the predominant genotype in Serbia and Slovenia and the detected PCV2 strains are closely related to those previously described in Europe and in other parts of the world.     

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 Na₂SeO₃/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.     

Further investigation of risk elements content in the bones of wild rodents from a polluted area in Slovakia

Background

Wild rodents are suitable for monitoring environmental pollution and exposure risk assessment for people living in contaminated areas. The content of selected risk elements in the femora of bank vole (Myodes glareolus), yellow-necked mouse (Apodemus flavicollis) and wood mouse (Apodemus sylvaticus) was estimated from the Kolíňany area of Slovakia, which is characterized by a high degree of environmental pollution. The rodents were trapped in February 2011 using standard theriological methods. All animals (n = 32) were adult males in good physical condition. The concentrations of Fe, Cu, Zn and Ni in their bones were determined by atomic absorption spectrophotometry.

Results

The highest concentrations of Fe and Cu were detected in the bones of yellow-necked mouse. Significant differences were observed for the content of Fe between A. flavicollis and M. glareolus (P < 0.05). The highest levels of Zn and Ni were found in the femora of wood mouse; however, significant differences were not detected between the rodents. Moreover, the concentrations of Cu, Zn and Ni were significantly higher in the bones of all three species (P < 0.05) in comparison with the values obtained in the same animal species at the same site in February 2007.

Conclusions

Our results demonstrate an increased accumulation of Cu, Zn and Ni in the femora of M. glareolus, A. flavicollis and A. sylvaticus from the Kolíňany area and thus indicate towards ongoing contamination of this locality.     

A new model for predicting the flight activity of Lobesia botrana (Lepidoptera: Tortricidae)

The grape vine moth, Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae), attacks vineyards mostly in Southern Europe and Northern Africa. The efficiency of most control methods depends on the treatment of pest populations at their most susceptible stages, therefore the prediction of the moth’s development cycle would help greatly in determining an optimal treatment schedule. One strategy for protecting against this pest is based on the day of maximum flight of males per generation (peak flight), when the mating between males and females and the laying of eggs reaches its maximum. The knowledge of these times would enable us to decide the kind of insecticide and the best time to target it. The time of maximum flight can be predicted by the Touzeau model of accumulated average daily temperature exceeding the development threshold. As shown, the fit of this model on data from the Ribera del Duero region in Spain is not accurate enough. Therefore, we propose an empirical and biologically plausible model based on logistic transformation of daily temperatures, which proves a better fit and permits simple inclusion of additional variables such as relative humidity.     

The effects of eggshell calcium (Biomin H®) and its combinations with alfacalcidol (1α-hydroxyvitamin D3) and menaquinone-7 (vitamin K2) on ovariectomy-induced bone loss in a rat model of osteoporosis

The purpose of this study was to investigate the impact of eggshell calcium (Biomin H^® dietary supplement) and its combinations with alfacalcidol (1α-hydroxyvitamin D₃) and menaquinone-7 (vitamin K₂) on ovariectomy-induced bone loss in rats. Adult female rats (n = 48) were divided into 6 groups of 8 individuals each: sham-operated rats (SHAM); ovariectomized (OVX) rats untreated; OVX rats treated with Biomin H^® (BIO); OVX rats simultaneously receiving Biomin H^®, vitamin D₃ (BIO + D₃); OVX rats simultaneously treated with Biomin H^®, vitamin K₂ (BIO + K₂) and OVX rats treated with Biomin H^®, vitamin D₃, vitamin K₂ (BIO + D₃ + K₂) during 8 weeks. Biochemical parameters, bone mineral density (BMD), bone mineral content (BMC) and femoral bone microstructure were determined. Plasma calcium and phosphate were increased in BIO + D₃ and BIO + D₃ + K₂ groups as compared to OVX. Alkaline phosphatase was elevated in OVX, BIO versus SHAM, BIO + D₃ + K2 groups. When compared to OVX group, decreased urine deoxypyridinoline was observed in all treated groups and femoral BMD, BMC were higher in BIO, BIO + D₃, BIO + D₃ + K₂ groups. The BIO + K₂ rats had similar densitometrical values than OVX individuals. Microcomputed tomography revealed increased trabecular relative bone volume (due to an increase in trabecular number) in BIO + D₃, BIO + D₃ + K₂ as compared to OVX. The higher relative bone volume in BIO + D₃, BIO + D₃ + K₂ groups was also accompanied by an increase in bone surface. In the cortical bone, an enhanced periosteal bone apposition was identified in BIO, BIO + D₃, BIO + K₂, BIO + D₃ + K₂ groups. The rats from BIO + D₃ + K₂ group had a higher area of primary osteon's vascular canals. In BIO + D₃, BIO + K₂, BIO + D₃ + K₂ groups, an increased area of secondary osteons was determined in comparison with OVX. Our results indicate the beneficial effect of triple application of Biomin H^®, vitamin D₃, vitamin K₂, as well as simultaneous administration of Biomin H^®, vitamin D₃ on the inhibition of ovariectomy-induced bone loss in a rat model of osteoporosis.     

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.     

Sex-related variations in bone microstructure of rabbits intramuscularly exposed to patulin

Background

Patulin, a toxic mold metabolite, has been found as natural contaminant of processed fruits, most notably apples, apple juices and other apple-based products. A number of adverse health effects in humans and animals are associated with patulin intoxication. The current study was performed to analyse possible toxic effects of patulin on femoral bone microstructure in adult rabbits in detail. Fourteen clinically healthy four-month-old rabbits of both sexes (6 males and 8 females) were included in the study. Animals from the experimental groups (group E♂, n = 3; group E♀, n = 4) were injected intramuscularly with patulin at dose 10 μg/kg body weight two times a week for 28 days. The dose of patulin was estimated based on the maximum permitted level of patulin for apple products intended for infants and young children. Three males and four females without patulin administration served as controls (groups C♂ and C♀). Cortical bone thickness and qualitative and quantitative histological characteristics of compact bone tissue were investigated.

Results

Intramuscular applications of patulin significantly increased the thickness of cortical bone in both sexes of rabbits. In patulin-exposed males, an absence of primary vascular longitudinal bone tissue near the endosteal border was observed, which could be associated with intensive bone remodeling. Femoral diaphyses of females displayed a lower number of secondary osteons in the middle part of the substantia compacta, and occurrence of the osteons near the periosteum. This could indicate alterations in bone turnover. Histomorphometrical evaluations showed significantly increased sizes of the primary osteons’ vascular canals (P < 0.05) in males exposed to patulin possibly due to mycotoxin-induced increased levels of testosterone.

Conclusions

This study demonstrates significant impact of intramuscular application of patulin on bone microstructure in adult rabbits. Moreover, we have found that the effects of patulin on qualitative and quantitative histological characteristics of compact bone are sex-dependent.     

Long-term peroral administration of bitter apricot seeds influences cortical bone microstructure of rabbits

Apricot seeds due to the presence of cyanogenic glycoside amygdalin belong to the popular “alternative cancer cures", although anticancer effect of amygdalin remains controversial. This in vivo study points to the effect of long-term peroral administration of bitter apricot seeds on bone microstructure of rabbits since chronic amygdalin toxicity in relation to bone parameters has not been investigated yet. Rabbits (n = 16) were randomly divided into four experimental groups of 4 animals each. Three experimental groups S1, S2 and S3 received commercial feed for rabbits mixed with crushed bitter apricot seeds at doses 60, 300 and 420 mg/kg bw during five months, respectively. The control (C) group received no apricot seeds. The long-term consumption of apricot seeds had no impact on total body weight, femoral weight and femoral length of rabbits. Also, microcomputed tomography (3D analysis) of cortical and trabecular bone tissues did not reveal any significant impact of amygdalin toxicity on relative bone volume, BMD, cortical bone thickness, bone surface, trabecular number, thickness, and their separation. On the other hand, histological (2D) analysis demonstrated evident changes in cortical bone microstructure consistent with a decreased density of secondary osteons in the middle part of substantia compacta due to a replacement of Haversian bone tissue by plexiform bone tissue, vasoconstriction in the primary osteons' vascular canals, Haversian canals, and decreased sizes of secondary osteons in rabbits from S1, S2 and S3 groups. These negative changes are associated with different vascularization and biomechanical properties of cortical bones.     

Exposure to non‐ionizing electromagnetic radiation of public risk prevention instruments threatens the quality of spermatozoids

The use of artificial insemination in cattle breeding has evolved to global extent, and insemination doses are often shipped via air transport which requires strict radiation‐based examinations. For the determination of effect of non‐ionizing radiation (NIR), to which are beings frequently exposed due to protection of airport or cultural event security, freshly ejaculated and cryopreserved bovine spermatozoa were used as experimental model. Following radiation with hand‐held metal detector in various exposition times (0, 10 s, 15, 30 and 60 min—groups FR, FR10, FR15, FR30 and FR60) the spermatozoa underwent motility and DNA fragmentation analyses. Study on cryoconserved semen treated with NIR was performed in time intervals 0, 10 s, 1 and 5 min (insemination doses radiated before cryoconservation—CB, CB10, CB1, CB5; samples radiated after freezing—CA, CA10, CA1 and CA5). Fresh semen and insemination doses radiated after cryoconservation showed significantly lower total and progressive motility. No effect on motility parameters was detected in semen extended with cryopreservative medium and radiated prior to freezing. Surprisingly, NIR showed a potential to stimulate spermatozoa velocity; however, the effect was modulated throughout the post‐thawing incubation. Based on the DNA fragmentation assay, sperm DNA stayed intact. Present study underlines the potential harm of NIR, which is frequently used in everyday life, with overall adverse impact on human and animal reproduction. Current study also points out on interesting short‐term spermatozoa stimulation induced by NIR.