Transfer of humoral and cell-mediated immunity via colostrum in pigs

The first aim of our study was to obtain information on the transmission of antigen-specific antibodies from colostrum to respiratory tract mucosa in piglets. The second aim was to confirm the biological relevance of the presence of lymphocytes in colostrum and the already described fact that these cells can penetrate the intestinal barrier and "colonize" peripheral blood and lymphatic tissues of piglets. Therefore, we performed an experiment in which sows were immunized with a model antigen Keyhole Limpet Hemocyanin and their piglets were euthanized at different intervals after birth and colostrum intake. Colostrum, bronchoalveolar lavage fluid and blood samples were collected for serological detection of antigen-specific antibodies. Lymphocytes isolated from peripheral blood and lymphatic tissues (mesenteric and tracheobronchial lymph nodes and spleen) of piglets were in vitro activated with the antigen. We found that colostrum-derived antibodies can cross into the respiratory tract mucosa. Furthermore, we found that antigen-specific lymphocytes were detectable in mesenteric lymph nodes and peripheral blood, but very rarely in spleen and tracheobronchial lymph nodes.     

Adaptation of aerobic training essentially involved autophagy,mitochondrial marker and muscle fibre genetic modulation in rat cardiac muscles

Information about the role of moderate acute treadmill training in modulating autophagy and mitochondrial markers that might be correlated with alteration of muscle fibre gene expression in rat cardiac muscles is very limited. In this present study, the researchers divided twenty male Wistar rats into four groups: sedentary control, 3, 6 and 15 days and subjected them to treadmill training with moderate intensity (20 m/min), 30 min each day. RNA was extracted from cardiac muscles and stored in temperature of −80°C. Specific primers were utilized for semi-quantitative PCR. Treadmill training decreased autophagy-related gene expression (LC3, p62) and upper stream signalling of autophagy (PIK3CA, Akt and mTOR) in 3 and 6 d, but stimulated gene expression of mitochondrial markers (PGC1α, Cox1, Cox2 and Cox4) in 15 days. αMHC gene expression increased while βMHC gene expression decreased in 15 days. In line with this, autophagy-related genes increased in 3 and 6 days and returned to baseline in 15 days. The increment in mitochondrial gene expression might be correlated with shifting gene expression of αMHC and βMHC in 15 days. Taken together, acute adaptation in cardiac muscles is stimulated by genetic modulation of autophagy, mitochondrial marker and muscle fibre that may explain physiological cardiac adaptation after training. This study can be used as a reference for optimizing performance in period of cardiac muscle adaptation stimulated by treadmill training.     

Subcutaneous ω-Conotoxins Alleviate Mechanical Pain in Rodent Models of Acute Peripheral Neuropathy

The peripheral effects of ω-conotoxins, selective blockers of N-type voltage-gated calcium channels (Ca_V2.2), have not been characterised across different clinically relevant pain models. This study examines the effects of locally administered ω-conotoxin MVIIA, GVIA, and CVIF on mechanical and thermal paw withdrawal threshold (PWT) in postsurgical pain (PSP), cisplatin-induced neuropathy (CisIPN), and oxaliplatin-induced neuropathy (OIPN) rodent models. Intraplantar injection of 300, 100 and 30 nM MVIIA significantly (p < 0.0001, p < 0.0001, and p < 0.05, respectively) alleviated mechanical allodynia of mice in PSP model compared to vehicle control group. Similarly, intraplantar injection of 300, 100, and 30 nM MVIIA (p < 0.0001, p < 0.01, and p < 0.05, respectively), and 300 nM and 100 nM GVIA (p < 0.0001 and p < 0.05, respectively) significantly increased mechanical thresholds of mice in OIPN model. The ED₅₀ of GVIA and MVIIA in OIPN was found to be 1.8 pmol/paw and 0.8 pmol/paw, respectively. However, none of the ω-conotoxins were effective in a mouse model of CisIPN. The intraplantar administration of 300 nM GVIA, MVIIA, and CVIF did not cause any locomotor side effects. The intraplantar administration of MVIIA can alleviate incision-induced mechanical allodynia, and GVIA and MVIIA effectively reduce OIPN associated mechanical pain, without locomotor side effects, in rodent models. In contrast, CVIF was inactive in these pain models, suggesting it is unable to block a subset of N-type voltage-gated calcium channels associated with nociceptors in the skin.     

Genetic diversity of Czech ‘Candidatus Phytoplasma mali’ strains based on multilocus gene analyses

In several European countries apple trees are affected by apple proliferation disease, which is usually associated with the presence of ‘Candidatus Phytoplasma mali’. During 2010, samples from several apple trees displaying proliferation symptoms were collected throughout the Czech Republic to verify identity of phytoplasmas detected in association with the disease. The majority of the 74 apple trees examined using molecular tools were positive for ‘Ca. P. mali’ presence. The 16S–23S ribosomal genes, the ribosomal protein genes and the nitroreductase and rhodonase like genes were then studied to verify phytoplasma strain variability on multigenic bases. Two RFLP profiles and correspondingly two genetic lineages were found in the PCR-amplified fragments covering the 16S–23S rDNA spacer region. ‘Ca. P. mali’ strains belonging to rpX-A subgroup were identified in the majority of the apple tree sampled, whereas phytoplasmas belonging to the rpX-B subgroup were distributed sporadically. The apple proliferation subtypes AP-15 and AT-2 exhibited nearly equal occurrence; the AT-1 subtype and a mixture of the two or all three of the AP subtypes were infrequently found. The PCR/RFLP results were confirmed by nucleotide sequence analyses of selected ‘Ca. P. mali’ strains.     

Effects of dietary supplementation of nickel and nickel-zinc on femoral bone structure in rabbits

Background

Nickel (Ni) and zinc (Zn) are trace elements present at low concentrations in agroecosystems. Nickel, however, may have toxic effects on living organisms and is often considered as a contaminant. This study reports the effect of peroral administrated Ni or a combination of Ni and Zn on femoral bone structure in rabbits.

Methods

One month-old female rabbits were divided into three groups of five animals each. Group 1 rabbits were fed a granular feed mixture with addition of 35 g NiCl₂ per 100 kg of mixture for 90 days. In group 2, animals were fed a mixture containing 35 g NiCl₂ and 30 g ZnCl₂ per 100 kg of mixture. Group 3 without administration of additional Ni or Zn served as control. After the 90-day experimental period, femoral length, femoral weight and histological structure of the femur were analyzed and compared.

Results

The results did not indicate a statistically significant difference in either femoral length or weight between the two experimental groups and the control group. Also, differences in qualitative histological characteristics of the femora among rabbits from the three groups were absent, except for a fewer number of secondary osteons found in the animals of groups 1 and 2. However, values for vascular canal parameters of primary osteons were significantly lower in group 1 than in the control one. Peroral administration of a combination of Ni and Zn (group 2) led to a significant decreased size of the secondary osteons.

Conclusions

The study indicates that dietary supplementation of Ni (35 g NiCl₂ per 100 kg of feed mixture) and Ni-Zn combination (35 g NiCl₂ and 30 g ZnCl₂ per 100 kg of the mixture) affects the microstructure of compact bone tissue in young rabbits.     

Direct effect of fertilization on microbial carbon transformation in grassland soils in dependence on the substrate quality

Response of microbial metabolism (growth, substrate utilization, energetic metabolism) to fertilization by N and P and resulting changes in soil‐organic‐matter (SOM) decomposition (priming effect) were studied in grassland soils with relatively high organic‐matter content. Treatments with and without glucose addition were studied to simulate difference between rhizosphere and bulk soil. Our expectation was that fertilization would decrease soil respiration in both treatments due to an increased efficiency of microbial metabolism. At first, fertilization activated microbial metabolism in both treatments. In glucose‐nonamended soils, this was connected with a short‐term apparent priming effect but if glucose was available, the higher energetic demand was covered by its mineralization in preference against SOM, causing significant SOM savings as compared to unfertilized soils. After a relatively short period of 1–3 d, however, the phase of deprived microbial metabolism occurred in both treatments, which was characterized by lower soil respiration in fertilized than in unfertilized soils. Fertilization further decreased net microbial growth following glucose addition, shortened turnover time of microbial biomass and changed the partitioning of assimilated glucose within microbial biomass (decreased accumulation of storage compounds and increased the proportion of mineralized glucose). As a result, fertilization reduced soil respiration mainly due to a deprivation of microbial metabolism. The rate and range of microbial response to fertilization and also the amount of saved soil C were larger in the soil with higher SOM content, likely driven by the higher content of microbial biomass.     

Mapping of urban environmentally sensitive areas in Bratislava city

Journal of Soils and Sediments - Environmentally sensitive areas within urban environment (U-ESA) play an essential role in maintaining urban ecological balance and ecological safety. The aims of...     

Characterization of <Emphasis Type="Italic">Agrobacterium</Emphasis> species by capillary isoelectric focusing

Tumorigenic and non-tumorigenic strains of Agrobacterium tumefaciens, A. rhizogenes, A.rubi, and A.vitis were examined using capillary isoelectric focusing, phenotypic determinative tests, PCR and fatty acid analysis. The isoelectric points (pI) of the 40 strains investigated clearly differentiated the strains according to their respective species. The different species were characterized with the following pI values: A. tumefaciens 2.2, A. rhizogenes 4.0, A. rubi 2.15, and A. vitis 2.6. This differentiation corresponded to the phenotypic, PCR and fatty acid characterizations. Strains with the similar chromosomal background but different plasmid content, e.g. A.vitis strain S4, and F2/5 gave the same pI values. Strains of Rhizobium species differed from Agrobacterium strains in their pI values. The advantage of capillary isoelectric focusing over the phenotypic determinative tests, PCR and fatty acid analysis is its speed (15 min), relative simplicity, and the very small amount of chemicals used. This rapid and simple method is a major improvement over the classical methods of separation of Agrobacterium species and should prove useful for rapid characterization of Agrobacterium-like colonies isolated from plant tumours for epidemiological and generic diversity studies.     

The inability of tapeworm Hymenolepis diminuta and fluke Dicrocoelium dendriticum to metabolize praziquantel

Biotransformation enzymes can, to a certain extent, protect parasitic worms against the toxic effects of anthelmintics and can contribute to drug-resistance development. The objective of our work was (1) to find and identify phase I and II metabolites of the anthelmintic praziquantel (PZQ) formed by the lancet fluke (Dicrocoelium dendriticum) and the rat tapeworm (Hymenolepis diminuta) and (2) to compare PZQ metabolites in helminths with PZQ biotransformation in rat as host species. Ultra high performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) was used for this purpose. During in vitro incubations, mitochondria-like and microsomes-like fractions (prepared from homogenates of adult worms or from rat liver homogenate) were incubated with 10 and 100 μM PZQ. Liquid/liquid extraction was used for samples during in vitro experiments. In the ex vivo study, living D. dendriticum and H. diminuta adults were incubated in RPMI-1640 medium in the presence of 50 nM or 100 nM PZQ for 24h. After incubation, the worms were removed from the medium and homogenized. Homogenates of worms, medium from the incubation of worms or rat hepatocytes and rat urine (collected during 24h after oral PZQ administration) were separately extracted using solid-phase extraction. The results showed that both D. dendriticum and H. diminuta enzymatic systems are not able to metabolize PZQ. On the other hand, thirty one different phase I and four phase II PZQ metabolites were detected in rat samples using UHPLC/MS/MS analyses. These results show that our experimental helminths, as the members of tapeworm and fluke groups of parasites, are not able to deactivate PZQ, and that the biotransformation enzymes of the studied helminths do not contribute to PZQ-resistance.