Reliability of three common fecal egg counting techniques for detecting strongylid and ascarid infections in horses.

The detection and quantification of nematode eggs using fecal egg count techniques have an irreplaceable role in equine parasitic control. The reliability, particularly precision and accuracy, of individual techniques have been described only for strongylid infections. The aim of this study was to compare three fecal egg count techniques used for the detection of the two most common equine nematode infections: strongylid and ascarid. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques were tested on spiked fecal samples with various levels of egg concentration (50, 100, 200, 500, 1000 and 3000 eggs per gram) and naturally infected mixed strongylid-ascarid samples with 30 replicates. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques had precision coefficients of variation of 44.33, 35.64 and 18.25% for the strongylid infection and 62.95, 35.71 and 18.95% for the ascarid infection, and percent accuracies (mean count/number of eggs spiked) of 97.53, 88.39 and 74.18% for the strongylid infection and 65.53, 83.18 and 90.28% for the ascarid infection, respectively. Accuracy depended greatly on the type of nematode, but precision did not. The Mini-FLOTAC technique was more precise than the Simple and Concentration McMaster techniques regardless of nematode type. Simple McMaster was the most accurate technique for detecting strongylid eggs, and Mini-FLOTAC was the most accurate technique for detecting ascarid eggs. Our results indicated that none of the current techniques were universally and sufficiently reliable for the simultaneous quantification of both of these common equine nematodes.

New mtDNA data reveal a wide distribution of the Japanese ginbuna Carassius langsdorfii in Europe.

In this study, records on the occurrence of the Japanese ginbuna Carassius langsdorfii from northern Germany, north-western Italy and southern Bosnia and Herzegovina are presented. The new findings, in addition to former studies reported in the Czech Republic and Greece, show that C. langsdorfii is much more widespread in Europe than was previously believed.

The first determination of Trichuris sp. from roe deer by amplification and sequenation of the ITS1-5.8S-ITS2 segment of ribosomal DNA.

Trichuris nematodes were isolated from roe deer (Capreolus capreolus). At first, nematodes were determined using morphological and biometrical methods. Subsequently genomic DNA was isolated and the ITS1-5.8S-ITS2 segment from ribosomal DNA (RNA) was amplified and sequenced using PCR techniques. With u sing morphological and biometrical methods, female nematodes were identified as Trichuris globulosa, and the only male was identified as Trichuris ovis. The females were classified into four morphotypes. However, analysis of the internal transcribed spacers (ITS1-5.8S-ITS2) of specimens did not confirm this classification. Moreover, the female individuals morphologically determined as T. globulosa were molecularly identified as Trichuris discolor. In the case of the only male molecular analysis match the result of the molecular identification. Furthermore, a comparative phylogenetic study was carried out with the ITS1 and ITS2 sequences of the Trichuris species from various hosts. A comparison of biometric information from T. discolor individuals from this study was also conducted.

Effect of lead in water on the absorption of copper, iron, manganese and zinc by sheep (Ovis aries) infected with sheep tapeworm (Moniezia expansa).

The sheep tapeworm (Moniezia expansa) and its host Ovis aries were analyzed by inductively coupled plasma optical emission spectrometry (ICP-OES) for their copper, iron, manganese, zinc and lead levels. Element concentrations in cestode parasites were compared to those in various organs (liver, kidney, and muscle) of sheep. Tapeworms in the small intestine of sheep that were administered 2g of Pb(CH(3)COO)(2) per os daily (7 days) had significantly higher lead concentrations than sheep tissues. Cu levels significantly increased after Pb administration in sheep muscle and sheep tapeworms. Contrarily, Zn content significantly decreased in sheep muscle, but significantly increased in sheep tapeworms. However, Mn content significantly decreased after Pb administration in sheep tapeworms. Furthermore, Fe content significantly decreased after Pb administration in sheep liver and kidneys.

Effect of Acanthocephalus lucii infection on total mercury concentrations in muscle and gonads of fish host (Perca fluviatilis).

The samples from 13 perches (Perca fluviatilis) - muscle with skin and bones; fish gonads; and acanthocephalan parasites were analysed for mercury (Hg). Hg concentrations were present in all analysed samples. There were found no statistically significant difference in Hg concentration in fish tissues between perches either with or without infection by the acanthocephalan parasite, Acanthocephalus lucii). In this study there was no evidence that acanthocephalan worms accumulate mercury from hosts. For this reason, A. lucii is not a suitable bioindicator for mercury pollution.

The initial discovery of thorny-headed worms in sheep.

Acanthocephalans belonging to the species Acanthocephalus lucii were found in the colon of a lamb from ecological farms in the Czech Republic. The main determination features used for these acanthocephalans are the shape and size of the hooks as well as the number of hooks on the proboscis. Three immature specimens measured 5.0-13.2mm in length; the appearance of the acanthocephalan body (in the studied material) suggests that passage through this unusual host causes the cystacanths to slightly increase in length. In the case of the lambs examined, the infection may be acquired through the accidental ingestion of the intermediate host - waterlouse (Asellus aquaticus).

Surface modification of cyclic olefin copolymers for osteochondral defect repair can increase pro-destructive potential of human chondrocytes in vitro.

Materials on the basis of cycloolefin copolymers (COC) are suitable for subchondral defect repairs. The objective of this study was to evaluate the influence of surface modification of COC and COC/LLDPE blends on the viability and gene expression of chondrocytes. Human chondrocytes were incubated on the surface of the studied materials. Half of the materials were plasmatically modified with a subsequent type II collagen application. The gene expression of matrix metalloproteinases (MMP-1,-3,-13), pro-inflammatory cytokines (IL-1, TNF-alpha) and apoptotic molecules (BAX, Bcl-2) was evaluated using quantitative Taq-Man PCR after 48 h incubation. Chondrocyte viability was evaluated by the MTT test after 2, 4 and 8 days of incubation. The synthesis of MMPs was measured by ELISA assay in cell culture medium after 48 h of incubation. Chondrocytes incubated on plasmatically modified in contrast to unmodified materials demonstrated significantly increased gene expression of IL-1 (p<0.05), MMP-1 and MMP-3 (p<0.05 for both comparisons) as well as MMP-13 (p<0.001). Increased gene expression was confirmed by significantly increased production of active forms of particular MMPs into the cell culture medium. Unlike surface unmodified polymers, the modified materials showed time-dependent reduction of chondrocyte viability. The gene expression of TNF-alpha and apoptotic molecules by chondrocytes was not significantly changed by different materials. Cycloolefin copolymers and their blends may represent suitable materials for tissue engineering, however, their surface modification followed by collagen type II application may, at least under in vitro conditions, reduce the viability of chondrocytes and induce their pro-destructive behavior. The potential benefit or disadvantage of surface modifications of materials for osteochondral defect repairs needs to be further elucidated.

Heavy metal accumulation in small terrestrial rodents infected by cestodes or nematodes.

The aim of the present study was to assess whether there is a difference in accumulation of heavy metal ions (Cd, Cr, Cu, Mn, Ni, Pb and Zn) in hosts (small mammals) infected by cestode parasites when compared to those without cestode infection. The abundance of gastrointestinal parasites and bioaccumulation of heavy metals in host livers and kidneys were measured. Contents of heavy metals in hosts were determined by ICP OES method. The hosts with cestode infection (Paranoplocephala sp.) had lower contents of heavy metals in their livers and kidneys compared to hosts with nematode infection (Mastophorus muris). The content of Pb, Cd, Cr, Cu and Ni was higher in kidneys than in livers, in both (cestode and nematode infected) rodents while the content of Mn was higher in livers. Content of Zn was similar. The content of heavy metals in host was decreasing with the increasing abundance of cestodes (Paranoplocephalo sp.). Species-response models to particular heavy metals are presented.

Biological and biochemical properties of the carbon composite and polyethylene implant materials.

We studied the biocompatibility of the carbon composites and polyethylene materials with and without collagen or collagen and proteoglycan cover. We used the in vitro technology to study the adhesion of model cells evalution, their metabolic activity and the production of TNF-alpha as a cytokine model. Under in vivo condition, the biocompatibility of tested polymers were studied in the implantation experiment, subcutaneously in the interscapular region in the laboratory rat. We have found in the in vitro assay favorable proliferation and the smallest production of pro-inflammatory TNF-alpha cytokine in cells adherent to the hydrophobic polyethylene material coated with biological macromolecules. Using in vivo tests performed by the implantation of materials to the rat we demonstrated that the materials are not cytotoxic. The tissue capsule surrounding the implants was not significantly influenced by the type of the implant and the pre-treatment by the biological molecules. However, the foreign-body giant multinucleated cells were observed only in the vicinity of the collagen - covered hydrophobic polyethylene implant. Interestingly, while the collagen coating improved the biocompatibility of tested polymers in vitro, the inflammatory reaction against this covered materials was higher under in vivo conditions. The pre-treatment of carbon composites by both types of biological macromolecules reduced the occurrence of carbon debris in the implantation site. The tested carbon composites and polyethylene materials are not toxic. The pre-treatment of the materials by extracellular matrix components increased their biological tolerance in vitro and reduced implant wears in animal experiment, which can be important for the medical application.

[New materials suitable for implantation--preparation for implants]. / Nové materiály vhodné pro implantáty--príprava implantátu.

PURPOSE OF THE STUDY: Connective tissue components, particularly collagen and proteoglycan, stimulate cell proliferation and thus promote tissue regeneration. This fact was utilized to test some new implant materials, which were covered by these components, for the speed and quality of their encapsulation. MATERIAL: The materials tested included polyethylene (PE) with either a hydrophobic (HPHO) or a hydrophilic (HPHI) surface and the C-C composite. Before implantation in experimental animals, the materials were coated with a thin layer of collagen-proteoglycan copolymer. METHODS: Collagen was obtained from calf hide (ISC 40 fraction) and proteoglycan was isolated from calf cartilage with 2M GuHCl. The concentrations of elements in implant materials were assessed by the method of ESCA photoelectron spectroscopy (electron spectroscopy for chemical analysis) and, using a comparison of the values of electron binding energies at inner levels with the published ones, the chemical states of elements were identified. RESULTS: Carbon, oxygen and silicon were identified on the surface of the PE HPHI sample; carbon, oxygen, calcium, manganese and sulfur were found on the PE HPHO sample and nitrogen, oxygen and fluorine were present on the C-C composite. DISCUSSION: In our group of patients there was a small rate of rheumatoid arthritis, while in most literature these patients are predominant. In accordance with the outcomes of other authors, ASS is fraught with recurrence. ASS does not strike to fibrous capsule, like an open surgery synovectomy, and from these islands synovialis could regenerate. CONCLUSIONS: The presence of manganese and calcium on the surfaces of the materials investigated indicates the existence of a collagen-proteoglycan copolymer, which contains NH2 and COO groups derived from collagen and SO4 groups from proteoglycan. It is necessary to verify this theoretical assumption based on measurement data in a biological experiment.

[Dependence of mechanical strength on the architecture of the compact bone of human shaft.]. / Závislost mechanické pevnosti na architektonice kompaktní kosti diafýzy u cloveka.

A secondarilly remodelled bone (the haversian bone) forming a wall of human shaft represents a significantly anisotropic material, which has different strengths in different directions. One of the factors influencing mechanical properties (strength in particular) of the bone is the architectonics of the compact bone. Osteons of the haversian bone are arranged in the direction of the dominant first principal stress and create two antirotational spiral systems in the opposite walls of the shaft. The aim of the work was to found out how the bone tensile strength depends on the orientation of osteons and check whether the premise applies that the architectonics of the haversian bone depends on the directions of the first principal stress and in what relation of strength moments the direction of the first principal stress corresponds to the actual orientation of osteons. The results of the tensile strength tests in wet samples from 10 femurs and the strength of the whole shaft in 9 pairs of femurs (cadaver material) have confirmed the premise that osteons are arranged in the directions of the dominant first principal stress and depend on the relation between bending and torsion moments. The strength of the femoral shaft is maximal in the physiological way of loading, i.e. in medial bending and outer rotation. The decisive motion on which the strength of the bone depends is torsion. In non-physiological loading the femoral shaft (by an opposite bending moment or opposite torsion moment in medial bending) the strength of the bone is significantly lower. From the biomechanical viewpoint the architecture of the haversian bone has an optimal and efficient structure. Key words: compact bone, bone strength, types of loading.

Spatial organization of the haversian bone in man.

The osteons in all human long bones are grouped in two helical antirotary systems of opposite oblique directions, situated in the contralateral walls of the diaphysis. Several arguments support the hypothesis that this special architecture arises from functional adaptations and that it depends on the orientation of the first principal stress. A new macroscopic method based on the filling of vascular canals of the undecalcified, polished bone with India-ink enabled us to study the spatial architecture of the haversian bone in the entire diaphysis. The osteon directions in normal and atypical femurs was compared with the direction of the first principal stresses, determined analytically in a cylindrical tube model of the diaphysis subjected to a combination of bending, torsion and compression. Under combined loading with the bending moment in the frontal plane to the medial side and with the torque moment in the sense of external rotation, the direction of the first principal stresses corresponded with the direction of osteons in the diaphysis of the femur. In both cases, the first principal stresses, as well as the osteons, were oriented in the opposite oblique direction in the medial and lateral walls of the model and of the bones. Between the two oblique fields a sharp boundary with an atypical organization of the principal stresses and osteons existed. In atypical femurs the osteonal orientation was longitudinal (likely unloaded femurs) or rotated 90 degrees (markedly anteriorly convex femurs). These observations support the hypothesis of a causal relation between the loading mode and the dominant osteonal direction. The organization of the haversian bone seems a typical example of functional adaptation.

Osteon orientation of the diaphysis of the long bones in man.

The orientation of osteons has been described in the long bones of man, using a special macroscopic method, which enabled the study of the osteonal architecture throughout the extent of the diaphysis. The osteons in all bones are arranged in two helical systems of opposite directions, which lie on the opposite sides of the diaphysis. The inclination of osteon direction from the bone axis in remarkably constant, varying between 5 degrees and 15 degrees. The boundary between the two fields is sharp. This special type of osteon orientation corresponds with the directions of the maximum principal stress and/or strain in the walls of the bones, which are exposed to both the bending and torque moments. The typical orientation of osteons reflects the history of loading of the bones in the course of physiological activity. The dominant stress state of every bone could be deduced from the osteon orientation and from the position of the pressure and tension fields on the surface of the bone.

[Structure of compact bone and types of stress in the long bones in humans]. / Struktura kompaktní kosti a zpusob namáhání dlouhých kostí cloveka.

The authors elaborated a new macroscopic method which made it possible to analyze the orientation of osteons in the entire diaphysis. All long bones of the human extremities were thus analyzed. In each bone the osteons are arranged in a specific pattern but according to a uniform plan. Most of them are oriented in an oblique direction and are arranged as two helices of opposite directions located on opposite sides of the bone. The two fields are separated by a sharp borderline. Biomechanical analysis revealed that the orientation of the osteons is consistent with the direction of the maximal principal stress which depends on the normal and torsional stress to which the bone is exposed. From the orientation of osteons the authors derived ex post the mode of dominant loading of all long bones: the femur is exposed to bending in the medial direction and to outer rotation, the tibia is exposed to bending in dorsal direction and to inner rotation. The humerus is bent in the medial direction, the radius in the dorsal one. Both these bones are exposed to loading in inner rotation. In the ulna and fibula the mode of stress could not be assessed by the authors' method. The results are summarized in two diagrams which indicate the orientation of osteons and the way in which the long bones in man are exposed to load. The authors discuss the theoretical and practical impact of these findings.