Band 6, Heft 2

Genetic analysis of resistance to infectious disease reveals many important cues that have led to new insights into the interaction between pathogen and host. This knowledge might help with a better prognosis for diseases, and to the development of novel therapeutics. This review focuses on genes and loci that control susceptibility to diseases with an important epidemiologic impact, such as AIDS, hepatitis B, gastritis and peptic ulcer, tuberculosis, leprosy, malaria, schistosomiasis and leishmaniasis. New perspectives for the integration of human and mouse genetics that contribute greatly to our understanding of regulatory mechanisms in health and disease, are also discussed.

Arthropods are very important organisms in the environment in relation to transmission of pathogenic infections to humans. The information on transmission of pathogenic infections to people by commercially offered food arthropods is scant. Consumption of seafood is a very popular dietary habit around the world. Whereas shrimp are the most commonly consumed seafood item, crabmeat has recently become a very popular commercial product, specifically for a majority of European countries. The transmission of waterborne protozoan parasites is facilitated by consumption of seafood harvested from contaminated waters, drinking water or via contact with recreational and surface waters, and remains common throughout the developing as well as the developed world. Protozoan infections pose a significant health risk for immunocompetent individuals, and may cause life-threatening diseases among immunocompromised and immunosuppressed individuals. The transmissive stages of human protozoan parasites are small in size and are shed in large numbers in feces of infected people and animals. These pathogens are resistant to environmental stressors (sometimes even to chlorine disinfection), and only a few of them (e.g., Cryptosporidium oocysts, Giardia cysts, or Vibrio bacteria) are able to cause infection in seafood consumers.

As the occurrence of Candida species infections increases, so does resistance against commonly-used antifungal agents. It is therefore necessary to look for new antifungal drugs. This study investigated the antifungal activity of recently isolated, synthesized and characterized antimicrobial α-helical amphipathic peptides (12–18 amino acids long) from the venom of hymenoptera (melectin, lasioglossins I, II, and III, halictines I and II) as well as a whole series of synthetic analogs. The minimal inhibitory concentrations (MICs) against different Candida species (C. albicans, C. krusei, C. glabrata, C. tropicalis and C. parapsilosis) of the natural peptides amounted to 4–20 µM (7–40 mg/l). The most active were the synthetic analog all-D-lasioglossin III and lasioglossin III analog KNWKK-Aib-LGK-Aib-IK-Aib-VK-NH2. As shown using a) colony forming unit determination on agar plates, b) the efflux of the dye from rhodamine 6B-loaded cells, c) propidium iodide and DAPI staining, and d) fluorescently labeled antimicrobial peptide (5(6)-carboxyfluorescein lasioglossin-III), the killing of fungi by the peptides studied occurs within minutes and might be accompanied by a disturbance of all membrane barriers. The peptides represent a promising lead for the development of new, effective antifungal drugs.

Propolis has been used in traditional folk medicine for ages owing to a number of biological effects. Four propolis samples of Czech and one of Slovak origin were extracted using Soxhlet apparatus and analysed by thin-layer chromatography. Raw propolis samples and their extracts were tested by microdilution broth method to determine minimal inhibitory concentration (MIC) in eight strains of human pathogenic fungi. Raw propolis samples showed a lower in vitro antifungal activity than their extracts. In general, the petroleum ether extracts exhibited the highest in vitro antifungal activity (MIC range of 16–64 µg/ml). The content of flavonoids in the samples varied according to region. The highest amount of flavonoids was found in sample A that originated from Broumov (4%). The most susceptible to the propolis extracts were Trichophyton mentagrophytes and Candida albicans. The propolis samples of Czech and Slovak origin and their extracts showed a considerable in vitro antifungal effect which was associated especially with nonpolar petroleum ether and toluene extracts. There was only a partial correlation between flavonoids content and in vitro antifungal activity.

The melanocortin 4 receptor (MC4R) is involved in the regulation of energy homeostasis and is known as one of the major hypothalamic regulators of food intake. Several studies have shown that replacement of aspartic acid at position 126 of the MC4R abolishes the ligand binding. We used the modified yeast Saccharomyces cerevisiae strain MMY28 to functionally express the MC4R and characterise the importance of this amino acid for ligand based activation of the receptor. The efficiency of the functional expression system was estimated by activation with αMSH, ACTH and THIQ and compared with cAMP response in mammalian cells. We generated the library of MC4R mutants randomised at the amino acid position 126. Recombinant MC4R clones were screened for the αMSH induced activity in yeast. From 9 different amino acids obtained only the natural aspartic acid displayed the ligand dependent activity of MC4R. The MC4R variants with glutamic acid and leucine at position 126, however, displayed higher background activity than other amino acid substitutions. The results suggest that the yeast expression system is suitable for screening of the MC4R receptor ligands and that the substitution of aspartic acid at position 126 of MC4R by different amino acids functionally inactivates the receptor.

The majority of mammals are unable to digest lactose due to post-weaning deactivation of the LCT gene, which is responsible for encoding the enzyme lactase (i.e., adult-type hypolactasia). A substitution of C with T at position −13910 bp upstream of the LCT gene has been linked to the lactase persistence phenotype in European populations. We investigated the frequency of LCT-13910C>T polymorphism in 223 blood donors from central Poland. All samples were genotyped by polymerase chain reaction and direct sequencing. The LCT-13910 T allele (lactase persistence) was present in 51% of individuals sampled from the Polish population. We did not find any non-European variants associated with lactase persistence (LCT-13907C>G, LCT-13913T>C, LCT-13915T>G), or any new polymorphisms within the sequenced region. Allele frequencies obtained are in agreement with results from other countries and confirm the unique pattern of distribution of the LCT-13910C>T genotype in Europe.

Asian sea bass immunoglobulin M (IgM) was purified from the sera of Lates calcarifer by affinity chromatography. Analysis of the purified IgM on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions revealed that the sea bass IgM was a tetrameric protein with a molecular weight of 896 kDa; it contained an equimolar heavy chain and light chain with molecular weight of 83 kDa and 27 kDa respectively. However, besides the covalently linked tetrameric IgM, noncovalently linked tetramer dissociated into dimeric and monomeric forms also demonstrated by non-reducing SDS-PAGE. Carbohydrate moieties were found to be linked with both heavy and light chains. A polyclonal rabbit anti-Asian sea bass IgM was prepared which showed a specific reaction of anti-fish IgM antibody with IgM of sea bass. Sea bass IgM concentration was determined in the serum by indirect ELISA. The average IgM concentration in the sera of the healthy sea bass was 5.4±1.8 mg ml−1; it amounted to 16.7% of the total serum protein.

The aim of the study was to characterize the 2,4-dichlorophenoxyacetic acid (2,4-D) degradative potential of three bacterial strains identified by MIDI-FAME profiling as Burkholderia cepacia (DS-1), Pseudomonas sp. (DS-2) and Sphingomonas paucimobilis (DS-3) isolated from soil with herbicide treatment history. All strains were capable of using herbicide as the only source of carbon and energy when grown in mineral salt medium (MSM) containing 2,4-D (50 mg/l). Over a 10 day incubation period, 69%, 73% and 54% of the initial dose of 2,4-D were degraded by strains DS-1, DS-2 and DS-3, respectively. Analysis of 2,4-dichlorophenol (2,4-DCP) concentration, the main metabolite of 2,4-D degradation, revealed that strains DS-1 and DS-2 may also have the potential to metabolize this compound. The percentage of 2,4-DCP removal was 67% and 77% in relation to maximum values of 9.5 and 9.2 mg/l determined after 4 and 2 days for MSM+DS-1 and MSM+DS-2, respectively. The degradation kinetics of 2,4-D (50 mg/kg) in sterile soil (SS) showed different potential of tested strains to degrade 2,4-D. The times within which the initial 2,4-D concentration was reduced by 50% (DT50) were 6.3, 5.0 and 9.4 days for SS+DS-1, SS+DS-2 and SS+DS-3, respectively.

Two solvent extraction procedures were used to investigate the extraction efficiency in terms of total antioxidant capacity and total phenols in apricot fruit. Samples were either sequentially extracted with aqueous ethanol (ethanol/water 80% v/v) and tetrahydrofuran or directly extracted with tetrahydrofuran. Each extract was analyzed for total antioxidant capacity by the Trolox Equivalent Antioxidant Capacity (TEAC) assay and total phenols by the Folin-Ciocalteu assay. The results showed that using sequential solvent extraction, the majority (85%) of the total antioxidant capacity and total phenols was due to hydrophilic compounds. In tetrahydrofuran direct extractions, the total antioxidant capacity and total phenols were higher than values obtained with aqueous ethanol and the sum of results obtained from sequential extracts for either total antioxidant capacity or total phenols was similar to the tetrahydrofuran-extract antioxidant values. A linear correlation between total antioxidant capacity and total phenols was found and was independent of the solvent extraction method. In conclusion, the choice of solvent is related to the antioxidant potential of fruit and depends on the food hydrophilic/lipophilic composition.

In order to evaluate the fruit maturity value of ‘Hayward’ kiwifruit (Actinidia deliciosa) as material for wine production, ripened and over-ripened fruit samples were analyzed. In addition, the effect of pectinase enzyme treatment on physicochemical characteristics of the wine during the fermentation process was also examined. The results showed that wine production from ripening kiwifruit took almost twice as long as from over-ripened fruit. The yield of wine production was increased from 63.35% to 66.19% by using riper kiwifruits and adding pectinase before amelioration. The soluble solid content (13.5%) in the wine produced from over-ripened fruits increased in comparison with the wine produced from ripened samples (10.3%). In both groups there was a decrease to 5° Brix at the end of fermentation. Difference of fruit maturity did not have a significant effect on acidy, pH, nor color of wine. The highest scores of total acceptance estimated during fermentation were for wine made from over-ripened fruits. Total phenolics content and antioxidant activities were similar in both wine groups while the mineral content, especially potassium, in wine made from over-ripened kiwifruit was higher than wine produced from ripened fruits. In conclusion, the quality characteristics of kiwifruit wine made from over-ripened fruit treated with pectinase showed higher values of wine in many aspects such as sensory value, alcohol and total phenolics content, antioxidant activity, minerals and production yield.

Esca is a destructive disease of the woody tissues of grapevine (Vitis vinifera L.) and due to the complexity of disease many aspects of host-pathogen interactions are not clearly understood. The histological characteristics of esca symptomatic petioles and internodes, collected from Cabernet Sauvignon and Sangiovese grapevine were studied. The tissues were fixed in FAA, dehydrated and embedded in Histoplast. To identify the lignified cell walls the sections were stained by Crystal violet and Erythrosin B and observed using an optical microscope. The main feature of tissues infected by esca disease was the minor lignification of vascular tissues, which was observed in petiole tissues before appearance of esca symptoms. The opportunity to utilize the histological examination of tissues as a method for the early detection of esca infections is hypothesized for the future application.

In a field experiment, the transfer of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) from contaminated soil to maize (Zea mays L.), sunflower (Helianthus annuus), poplar (Populus nigra × P. maximowiczii) and willow (Salix × smithiana) and the distribution of PCB congeners in maize and sunflower was investigated. The former waste incinerator in Hradec Králové (Czech Republic) was chosen for the experiment. Results of plot screening showed heterogenous contamination by PCBs and PAHs. PCB soil contamination was evidently caused by Delor 106 or Aroclor 1260 stocking and PAH contamination by chemicals containing fluoranthene, benzo/b/fluoranthene, phenanthrene and pyrene. Tested plants were planted on a contaminated field site, in soil contaminated with 1530 µg/kg of total PCBs and 0.138 and 3.42 mg/kg of total PAHs. The results show that maize and sunflower roots accumulated the most PCBs from soil. These plants accumulated hexa- and heptachlorobiphenyl congeners more than tri-, tetra-, and pentachlorobiphenyl congeners. Total concentrations of PAHs in tested plants ranged from 0.096 to 1.34 mg/kg. The highest phenanthrene concentration was found in aboveground biomass of sunflower and the highest concentration of pyrene, in maize roots.

Bryophytes are abundant in streams and are a habitat for many invertebrates, but their contribution to the diet of fluvial zoobenthos is still debated. To estimate the amount of bryophyte-derived organic matter assimilated by benthic invertebrates, we used a combination of fatty acid and stable isotope analyses during a four-year monthly study of a littoral site in the Yenisei River (Siberia, Russia). Acetylenic acids, which are highly specific biomarkers of the water moss Fontinalis antipyretica, were found in lipids of all dominant benthic animals: gammarids, ephemeropterans, chironomids and trichopterans. The dominant zoobenthic species, Eulimnogammarus viridis, had maximum levels of the biomarkers in its biomass during winter, and minimum levels in summer. The zoobenthos in the studied site regularly consume and assimilate bryophyte-derived organic matter as a minor supplemental food. This consumption increases in winter, when the main food source of the zoobenthos, epilithic biofilms, are probably scarce.

Comparative analysis of algal communities in the rivers of Israel was completed to highlight the influence of environmental variables on biodiversity. The study revealed that 671 species of algae and cyanobacteria belonging to nine taxonomic divisions were present during 2002–2009 in the Yarqon, Alexander, Hadera, Qishon, Oren, Lower and Upper Jordan, and Zin rivers. The species richness of each river was evaluated by taxonomic structural comparison, geobotanical, hierarchical cluster analysis, and the degree of relatedness for different levels of taxonomic resolution. The analysis revealed close similarity of the Upper Jordan and Oren rivers. The average taxonomic distinctness index showed that the Yarqon, Oren, Upper Jordan, and Qishon communities were partly degraded due to permanent environmental disturbances. The variation in taxonomic distinctness index showed that the Alexander, Yarqon and Hadera communities were formed not only due to anthropogenic factors but also through long-term climatic impact. The most abundant indicator species inhabit low streaming and standing alkaline waters of medium salinity and low to medium organic pollution. The statistical approaches allowed discrimination between climatic and anthropogenic factors that impact upon the riverine biodiversity in semi-arid environments. Analysis shows the influence of anthropogenic factors was strongly modulated by climatic impacts causing a marked decease of species richness from north to south.

Serpentine substrate in Serbia covers wide areas mainly in Kosovo and in central, western and south-western part of the country. On the serpentines of Serbia, particularly on the skeleton soils of hills and on lower elevation mountain slopes, there are some xerophilous and steppe-like vegetations. These xerophilous grasslands include mostly Pontic, Pontic-Submediterranean and/or Mediterranean-Submediterranean-Pontic plants. The distribution of 161 taxa of Pontic, Pontic-Submediterranean and Mediterranean-Submediterranean-Pontic areal — types on serpentinites of Serbia was analyzed in order to distinguish centers of richness and diversity. The distribution of taxa was analyzed with respect to geographic, ecological and climatic factors. The qualitative composition of Pontic flora on the serpentine areas was also compared. The greatest floristic richness and the center of diversity of Pontic flora on serpentine of Serbia was recorded in central part of the area investigated.

This study is the first to analyze the relationship between ecological indices (substrate moisture, nutrient content, substrate dispersion/aeration, substrate pH and humus content) and life forms of the vascular flora in and around Lake Provala; these relationships may serve as reliable indicators of the ecological conditions prevailing in this ecosystem. Since the development of certain life forms, in addition to climatic conditions, depends on plant requirements for several major environmental factors expressed as ecological indices, we established the relationship between the ecological index and life form of vascular plants collected over an eight year period in this system using correspondence analysis. We found a significant correlation between the development of certain life forms and levels of substrate moisture, nutrient content and substrate dispersion/aeration. These relationships help explain the predominance of hemicryptophytes and hydro-helophytes in the riparian zone of the lake, as these forms are perfectly adapted to water-saturated or aquatic environments rich in nutrients and relatively well aerated. There was no significant relationship between life forms and substrate pH or the content of organo-mineral compounds (humus) in the soil.

Research on the feeding habits of mammalian carnivores relies mainly on the analysis of stomach contents and faecal samples, but the outcomes of these two methods have only been compared in a few studies, with contrasting conclusions. In an Alpine area of NW Italy, we analysed both fox faeces collected along standardised transects, and the stomach contents of road-killed individuals. Faecal analysis involved the identification of macroscopic fragments, the identification of earthworm chaetae, and the assessment of relative volumes using Kruuk and Parish’s technique. Use of both methods indicated that the diet of the red fox included mainly fruit and mammals, but quantitative differences emerged. Garbage, birds, and cultivated fruit were overrepresented in the stomach contents, while earthworms, mammals, and wild fruit prevailed in the faecal samples. Logistic Regression Analysis suggested that the method of analysis was the main factor in determining the occurrence of food items in fox diet. Nonetheless, evidence suggests that road-killed foxes may include a disproportionately high percentage of synanthropic individuals and therefore be biased towards anthropogenic food. Results suggest that by using Kruuk and Parish’s technique, the main limitation of faecal analysis, i.e. the inaccurate estimation of the relative volume of each food item, can be overcome.

Counter-adaptations of predators towards their prey are a far less investigated phenomenon in predator-prey interactions. Caudal autotomy is generally considered an effective last-resort mechanism for evading predators. However, in victim-exploiter relationships, the efficacy of a strategy will obviously depend on the antagonist’s ability to counter it. In the logic of the predator-prey arms race, one would expect predators to develop attack strategies that minimize the chance of autotomy of the prey and damage on the predator. We tested whether avian predators preferred grasping lizards by their head. We constructed plasticine models of the Italian wall lizard (Podarcis sicula) and placed them in natural habitat of the species. Judging from counts of beak marks on the models, birds preferentially attack the head and might also avoid the tail and limb regions. While a preference for the head might not necessarily demonstrate tail and limb avoidance, this topic needs further exploration because it suggests that even unspecialised avian predators may see through the lizard’s trick-of-the-tail. This result may have implications for our understanding of the evolution of this peculiar defensive system and the loss or decreased tendency to shed the tail on island systems with the absence of terrestrial predators.