Volume 68, Issue 6

Luminous vibriosis is a shrimp disease that causes major economic losses in shrimp industry as a result of massive shrimp kills due to bacterial infection caused by Vibrio species. Use of antibiotics to control Vibrio in shrimp aquaculture is not allowed in the United States and so it is necessary to develop an alternative pathogen control method for shrimp production. Short-chain fatty acids have been used as food preservatives for a long time. Organic acids are commonly added in feeds in animal production, such as chicken, pig, and cattle. In this study, growth inhibition effects of formic acid on five selected Vibrio species, namely Vibrio alginolyticus, Vibrio cholerae, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus were studied. The Vibrio bacteria were grown on both solid and liquid media using Muller-Hinton agar and alkaline peptone water, respectively, with various concentrations of formic acid. Bacterial growth was monitored in the liquid media using optical density method. The results showed significant inhibition of growth of all five Vibrio species by formic acid at low concentration. The effective concentration (EC50) values were calculated for all five Vibrio species, which were less than 0.039% of formic acid. The results are encouraging to supplement formic acid in the shrimp feed as a control mechanism to reduce Vibrio outbreak in shrimp aquaculture system.

Xylanases constitute an important industrial enzyme, which hydrolyzes the polysaccharide xylan. In this work, a novel Streptomyces strain producing cellulase-free xylanase was isolated from the soil samples collected from the mangrove forest of Kadalundi, Kerala, India. The strain produced unique enzyme, which exhibited optimal activity at pH 9.0 and tolerance up to pH 12.0. Media engineering was carried out to improve the enzyme production, which showed best enzyme production at 30°C, medium pH 9.0 and incubation time of 48 h. Enzyme was highly thermo-tolerant up to 70°C and alkaline tolerant. Partial gene amplification as well as partial purification of enzyme was carried out to characterize the enzyme. The unique features of the enzyme make it an ideal candidate for industrial application for paper and pulp industry.

Microbial inulinases are an important class of industrial enzymes, which are used for the production of fructooligosaccharides and high-fructose syrup. Endoinulinase producing bacterial strains were isolated from soil samples taken from the vicinity of Asparagus sp. root tubers. All the bacterial strains were screened for inulinase activity. The primary screening was carried out based on hydrolytic zone on agar plates containing inulin-based medium and Lugol’s iodine solution. Thus 30 inulinase producing bacterial strains were isolated. Out of 30 strains, 5 bacterial strains were found endoinulolytic, whereas 25 were exoinulolytic on the basis of action pattern of the enzyme. In tertiary screening, the bacterial isolate AS-08 was found to be most efficient for inulinase activity. Morphological, biochemical and physiological characteristics of the bacterial isolate AS-08 confirmed it as Bacillus sp. Furthermore, species-specific identification by 16S rDNA sequencing and phylogenetic analysis revealed the isolate as Bacillus safensis. Bacillus pumilus SH-B30 was found to be the nearest homolog. The strain showed maximum inulinase activity (12.56 U/mL) after 20 h of incubation at 37°C.

Lectins are non-immune carbohydrate-binding proteins or glycoproteins with specific binding sites for certain glycoconjugates. Fungal lectins have been documented for their antitumour, antiproliferative, immunomodulatory, hypotensive and insecticidal effects. In the present study, a mycelial lectin having molecular mass 55 kDa has been purified and characterized from Lentinus squarrosulus. Biological action spectrum of the lectin revealed agglutination of all human blood types (A, B, O, AB), goat, sheep, rabbit and pig erythrocytes. Neuraminidase treatment of blood type O erythrocytes considerably augmented hemagglutination titre. Carbohydrate inhibition studies showed its high affinity to mucin and asialofetuin. Lectin was purified by a combination of ammonium sulphate precipitation, dialysis, ion exchange chromatography and gel filtration chromatography. Optimum pH for lectin activity was observed to be 6.5–8.0 and optimum temperature was 25–30°C. Lectin showed poor pH stability and was stable within pH 7.0–7.5. It was highly thermostable and could withstand temperature upto 70°C. Lectin activity was sensitive to ethylenediaminetetraacetic acid and denaturants.

As majority of chemical compounds identified as anti-cancerous are toxic to normal cells, the discovery and identification of new safe drugs is a necessity in the biomedical field. The antioxidant, antitumour and immunomodulating properties of an exopolysaccharide of sequence -α-d-glucose, α-d-mannose and β-d-glucose-, purified from a probiotic Lactobacillus plantarum was studied. The immunostimulation of the compound in human lymphocytes was seen at a concentration of 0.1 mg/mL with 20% proliferation rate. The antitumour studies by morphological apoptosis determination and 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay on breast adenocarcinoma cell line (MCF-7) exhibited an IC50 of 10 mg/mL for the compound.

Silver nanoparticles (AgNPs) have attracted the attention of researchers because of their unique properties and applications in various fields, such as medicine, catalysis, textile engineering, and pollution treatment. The green synthesis of AgNPs has many advantages, such as less time requirement, highly stable AgNPs, better control over crystal growth, morphology, ease for scale up, and economic viability. Syzygium aromaticum (clove) was used for the extracellular biosynthesis of AgNPs. Eugenols are the active biomolecules present in clove, responsible for the bioreduction of AgNO3 (Ag+) leading to the formation and capping of AgNPs (Ag0). One molecule of eugenol releases two electrons and these two electrons will be taken by 2 Ag+ ions and these will get reduced to 2 Ag0. The synthesis of AgNPs was confirmed by the appearance of brown colour. The synthesized AgNPs were characterised by various techniques, such as UV-VIS spectroscopy, transmission electron microscopy, X-ray diffraction and Fourier transformed infrared spectroscopy. The synthesised AgNPs have λ max of 440 nm. It was evaluated that the AgNPs were biphasic in nature (cubic + hexagonal) with an average size of 50.0 nm. The synthesized AgNPs showed significant antimicrobial activity against Bacillus cereus NCDC 240 as they are nano-sized and have high surface area to volume ratio. AgNPs inhibit the growth of bacteria by various ways, such as by disrupting the cell membrane of bacteria, uncoupling the oxidative phosphorylation, inhibiting the DNA replication, forming free radicals and affecting the cellular signalling of bacteria leading to cell death.

Zymomonas mobilis, a Gram-negative ethanologenic non-pathogenic bacterium, is reported to exhibit resistance to high concentrations of β-lactam antibiotics. In the present study, Z. mobilis was found to be resistant to I-IV generations of cephalosporins and carbapenems, i.e. narrow, broad and extended spectrum β-lactam antibiotics. We have analysed the genome of Z. mobilis (GenBank accession No.: NC 006526) harbouring multiple genes coding for β-lactamases (BLA), β-lactamase domain containing proteins (BDP) and penicillin binding proteins (PBP). The conserved domain database analysis of BDPs predicted them to be members of metallo β-lactamase superfamily. Further, class C specific multidomain AmpC (β-lactamase C) was found in the three β-lactamases. The β-lactam resistance determinants motifs, HXHXD, KXG, SXXK, SXN, and YXN are present in the BLAs, BDPs and PBPs of Z. mobilis. The predicted theoretical pI and aliphatic index values suggested their stability. One of the PBPs, PBP2, was predicted to share functional association with rod shape determining proteins (GenBank accession Nos. YP_162095 and YP_162091). Homology modelling of three dimensional structures of the β-lactam resistance determinants and further docking studies with penicillin and other β-lactam antibiotics indicated their substrate-specificity. Semi-quantitative PCR analysis indicated that the expression of all BLAs and one BDP are induced by penicillin. Disk diffusion assay, SDS-PAGE and zymogram analysis confirms the substrate specificity of the β-lactam resistance determinants. This study gives a broader picture of the β-lactam resistance determinants of a non-pathogenic ethanologenic Z. mobilis bacterium that could have implications in laboratories since it is routinely used in many research laboratories in the world for ethanol, fructooligosaccharides, levan production and has also been reported to be present in wine and beer as a spoilage organism.

Bacosides, constituents of Bacopa monnieri (Linn.), are reported to be potential therapeutic saponins in the cure of Parkinson’s disease (PD). However, detailed mechanism for control of PD by bacosides is not well documented. PD has been reported to be caused by genetic mutations in leucine-rich repeat kinase 2 (LRRK2) leading to higher kinase activity that has been identified as a major cause of familial PD. The LRRK2 was thus proposed as an important marker in the pathogenesis of PD. This suggests that inhibition of LRRK2 holds promise as a potential treatment for PD. Our study focuses on the possible application of bacoside A constituents as potential inhibitors of LRRK2. In this work, we have carried out the in silico molecular docking studies of bacoside A constituents with LRRK2, proposing their role as an inhibitor in PD. The study has revealed the significant interactions between bacosaponin and LRRK2 having ten H-bonds at receptor-ligand site with binding affinity −7.5 kcal/mol. Hence, amongst the studied triglycosidic saponins, bacosaponin was analyzed to be a better ligand, proposing it to be a major constituent in inhibiting enzymatic activities of mutated LRRK2.

In the present study, Lactobacillus ingluviei ADK10 (Acc. No. JQ395039) from intestinal origin was tested for its probiotic characteristic as well as uremia ameliorating activity on acetaminophen induced uremic rats. The results revealed that L. ingluviei ADK10 was able to tolerate pH 3.0–9.0 and 0.5% bile salt along with good hydrophobicity (67%) and adherence index with Ht-29 cell line on 258/100 cells. It was susceptible to 20 antibiotics. The organism was able to degrade food ingredients, like starch and milk proteins. The strain showed significant growth inhibition of Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Shigella dysentery, Pseudomonas aeruginosa, and Klebsiella pneumoniae (average diameter of 10 mm). The therapeutic potentiality of this probiotic bacterium was tested against acetaminophen induced uremic rats. It was found that supplementation of L. ingluviei ADK10 for 14 days with food reduced severe increase of uremic profiles, such as blood urea (85%), creatinine (68%) and uric acid (41%) in comparison to the uremic rats. Moreover, during the feeding of rats with probiotic strain at a dose of 1×109 bacteria, reduction of enterobacteria in faeces was observed. Our studies indicated that L. ingluviei ADK10 could be used as a health-promoting probiotic along with antiuremic efficacy.

Mining of natural sources for new secondary metabolites has a successful history, which is reflected by the fact that over 50% of all drugs, currently on the market, are derived from natural products. Bacteria are one of the most important sources of bioactive natural products destined for drug discovery. However, less than 1% of the microorganisms observed in different habitats have been cultivated and characterized. To explore the genomic and functional diversity of the vast majority of the microbial world, novel methods were introduced, which are based on analysis of a DNA isolated from environmental communities. Metagenomics represents a strategy offering access to the genetic information present in uncultured bacteria by screening of libraries constructed from DNA isolated from different habitats. Functional- and sequence-driven screens are the major approaches employed to mine metagenomic libraries. This review aims to highlight discoveries in this area and discusses the possible future directions of the field.

After soil surface disturbances biological soil crusts (BSC) cover rapidly the topmost soil millimeters. Depending on BSC age, development of soil water repellency, water infiltration and soil surface stability are influenced by this thin surface sealing. Within this study disturbed, early- mid- and late successional stages of BSC development were examined along a recovery transect. The results show an increase in water repellency and a decrease in water sorptivity and conductivity with ongoing BSC succession. Penetration resistance data shows very stable thin surface protection by cyanobacteria in early successional BSC that is non-repellent. Later successional stages show increased water repellency and lower water conductivity. We conclude that BSC development induces changes in surface structure and wettability. The soil surface wettability is strongly linked to the BSC community composition.

Transport of radioactive iodide 131I− in a black clay loam soil under spring barley in an early ontogenesis phase was monitored during controlled field irrigation experiment. It was found that iodide bound in the soil matrix could be mobilized by the surface leaching enhanced by mechanical impact of water drops and transported below the root zone of crops via soil cracks. The iodide transport through structured soil profile was simulated by the one-dimensional dual-continuum model, which assumes the existence of two inter-connected flow domains: the soil matrix domain and the preferential flow domain. The model predicted relatively deep percolation of iodide within a short time, in a good agreement with the observed vertical iodide distribution in soil. The dual-continuum approach proved to be an adequate tool for evaluation of field irrigation experiments conducted in structured soils.

Topsoil macropores of two plots under no-tillage and conventional tillage were analyzed. A leguminous-cereal rotation was applied for six cycles under dry-land farming system (crop residues were removed). The clay-loam soil shows some vertic characteristics. The main goal is to identify the relationship between the top soil macro and meso-pore distribution for the two tillage systems (at the end of sixth cycle of cultivation) with the annual crop production (rainfall in normal growing period and crop production values are included). Unaltered topsoil samples were taken from 0 to 60 mm (row and interrow positions) and from the immediate depth (60 to 110 mm) in both plots (conventional and no-tillage). The morphometric analyses of 66 polished slices were carried out with the aim to identify differences in soil macro and meso-pore organisation. Soil macropores were classified by size (area) and elongation ratio and by form factor and equivalent pore diameter. No appreciable differences were observed. Soil macro and meso-pore distributions of samples were also compared. The main difference observed between topsoil’s treatments was a different macropore size distribution between topsoil positions. The presence of larger macropores was higher in conventional tillage compared to no-tillage. Samples taken from row and deeper positions of conventional tillage show a somewhat higher amount of macropores in the range between 2 to 2.3 mm equivalent pore diameter. Soil macropores contribute to increase soil aeration and soil drying when topsoil is too wet in critical periods of crop development. Conventional tillage (crop residues removed), provides to the topsoil of a larger lateral and vertical variability of macropore distribution than no-tillage topsoil.

Pines, used for sand dune stabilization, can influence the hydrophysical parameters and water flow in an aeolian sandy soil considerably, mainly due to soil water repellency. Two sites, separated by distance of about 20 m, formed the basis of our study. A control soil (“Pure sand“) with limited impact of vegetation or organic matter was formed at 50 cm depth beneath a forest glade area. This was compared to a “Forest soil” in a 30-year old Scots pine (Pinus sylvestris) forest. Most of the hydrophysical parameters were substantially different between the two soil surfaces. The forest soil was substantially more water repellent and had two-times the degree of preferential flow compared to pure sand. Water and ethanol sorptivities, hydraulic conductivity, and saturated hydraulic conductivity were 1%, 84%, 2% and 26% those of the pure sand, respectively. The change in soil hydrophysical parameters due to soil water repellency resulted in preferential flow in the forest soil, emerging during a simulated heavy rain following a long hot, dry period. The wetting front established in pure sand exhibited a form typical of that for stable flow. Such a shape of the wetting front can be expected in the forest soil in spring, when soil water repellency is alleviated substantially.

Variability in soil moisture on a steep slope near a ridge in a forested mountain range, Shikoku, Japan, was studied observationally and numerically. Vertically integrated soil moisture, from a depth of −60 cm to the surface, W, was introduced as a key indicator, and its seasonal variation was analysed on a daily basis from August 2011 to August 2012. The “bucket with a bottom hole” (BBH) model of Teshima et al. (2006) was improved to consider the forest environment in simulating the variation in W. A “big-leaf” model was incorporated into the modified BBH model to estimate transpiration and interception by trees. The simulated soil moisture agreed reasonably with observed values on a daily to inter-seasonal timescale.

Salinization of crop fields is a pressing matter for sustainable agriculture under desertification and is largely attributed to root absorptive functions of the major crops such as maize. The rates of water and ion absorption of intact root system of maize plants were measured under the salinized condition, and the salt absorptive function of maize roots was analyzed by applying different two kinetic models of root ion absorption (i.e. the concentration dependent model and the transpiration integrated model). The absorption rates for salinization ions (Na+, Cl−, Ca2+ and Mg2+) were found to depend on ion mass flow through roots driven by the transpiration, and therefore the transpiration integrated model represented more accurately rates of root ion absorption. The root absorption of salinization ions was characterized quantitatively by two model parameters of Q′max and K′M involved in the transpiration integrated model, which are considered to relate to the potential absorbing power and the ion affinity of transport proteins on root cell membranes, respectively.

The paper focuses on the evaluation of transpiration as a physiological process, which is very sensitive to drought stress. Reactions of 25-year-old Norway spruce (Picea abies (L.) Karst.) trees to drought were examined during 2009 summer. Sap flow rate (SF), meteorological and soil characteristics were measured continually. Vapour pressure deficit of the air (VPD) and cumulative transpiration deficit (KTD) was calculated. During the second half of the vegetation period, the decrease in soil water content was observed and irrigation was applied to a group of spruce trees, while the second group was treated under natural soil drought. On the days, when the differences in transpiration between irrigated (IR) and non-irrigated (NIR) trees were significant (21 days), transpiration of NIR trees was only 23% of the transpiration of IR trees. We found significant differences in transpiration when the soil water content (SWC) of NIR variant at a depth of 5–15 cm ranged from 10.4 to 13.7%. Under both regimes of water availability, daily transpiration significantly responded to atmospheric conditions. However, the influence of all assessed meteorological parameters on SF of NIR trees was significantly lower than on IR tree. The dependency of transpiration on evaporative demands of atmosphere decreased with the decreasing soil moisture. Cumulative transpiration deficit of the stand during the entire evaluated period was 50.9 mm. The difference between the transpiration of the mean NIR tree and of the mean IR tree was 278.8 L over the assessed period of 47 days (5.9 L per day). The transpiration of NIR trees was 40.3% from the transpiration of IR trees during this period.

The main components of drought tolerance of six maize genotypes were studied to evaluate crop performance in water limiting environments: (1) the postponement of dehydration by reduced transpiration rate (TR) and an increased efficiency of water acquisition from soil; (2) the tolerance of dehydration by effective physiological water use. The aim was to describe the genotype dependent response to drought in leaf and root traits and water relations using data from controlled environment and field experiments, and using dynamic simulation by the Swedish Coup model. High genetic variation was detected in the root density, acquisition efficiency and water use among the genotypes. The female parent lines had the greatest TR with the smallest dry matter accumulation in water deficiency, whereas hybrids could acquire more water from dryer soil while maintaining a lower TR. Hybrid Mv 444 increased water potential more strongly in leaves than hybrid Norma. The postponement of dehydration was observed for Norma, while more tolerance to dehydration characterized Mv 444. Simulation was an effective tool for testing hypotheses considering water acquisition efficiency and for summarizing the results of the measurements in a formalized structure; it helped to quantify the dynamics of water availability and the impact of drought on the growth of the maize genotypes.

Residue retention and no-till farming have been widely adopted to reduce erosion risk, but residue retention in particular is becoming less common due to issues with weed control, and competing markets for residue such as bioenergy production. For this reason, the impact of residue removal on soil water contents in a sandy soil in a Mediterranean-type environment was evaluated. Crop residues were removed by burning or conventional tillage annually in autumn (April or May) from 2008 until 2011. Surface residue cover and soil water contents were measured in summer (February-March) every year from 2008 until 2012, at the time of minimum soil water content (approaching air-dry). After three years of residue removal, average ground cover in the subsequent summers (2011 and 2012) decreased from 78% to 51%, and surface soil water contents decreased from 5.1% to 3.1%. Tillage also significantly decreased ground cover (from 72% to 58%) and soil water (from 4.2% to 3.9%) during the same time period. Changes in surface cover and soil water content indicate that residue removal will have implications for soil health and sustainable crop production.

Woody encroachment affects the biohydrology of rangelands worldwide and can increase evapotranspiration by increasing plant rooting depth, increasing the duration of the growing season, or by initiating a process of hydrologic recovery in formerly overgrazed landscapes. Eastern redcedar (Juniperus virginiana) is encroaching rapidly into rangelands in the Southern Great Plains of the USA, and beyond, including Oklahoma. However, the degree to which increasing growing season duration causes higher evapotranspiration after encroachment is not known. Here we show that increasing the duration of the growing season in north-central Oklahoma’s water-limited climate from seven months (April–October) to 12 months increases modeled evapotranspiration only marginally, from 95% to 97% of precipitation. However, this increase in evapotranspiration with woody encroachment into grassland corresponded to a two-thirds reduction in deep drainage. This study’s estimate of the hydrologic effects of eastern redcedar encroachment is likely to be highly conservative because it does not take into account the runoff-inducing effects of livestock grazing. Comparing simulated hydrologic fluxes in the present study to past work measuring runoff from grazinglands suggests that eastern redcedar encroachment into overgrazed rangelands is likely to increase evapotranspiration significantly. Whether or not eastern redcedar encroachment effects on evapotranspiration are discernable at the watershed scale will depend on the extent of encroachment throughout the watershed. Further research is necessary to quantify how the hydrologic effects of eastern redcedar encroachment vary due to climatic gradient.

The freshwater bryozoan species Pectinatella magnifica was found in 6 sandpits and in 19 mostly extensively managed ponds in the Protected Landscape Area and Biosphere Reserve Třeboňsko (Czech Republic) from its first record (in 2003) to 2012. Mean fresh biomass and abundance of P. magnifica colonies were 0.6 ± 1.5 kg m−2 and 0.7 ± 1.1 colony m−2 (± SD), respectively, in the shoreline zone during the growing season 2006–2011. The maximum biomass was mostly recorded during the first half of August in all basins. Colonization of further localities was recorded rather than increasing of P. magnifica biomass or abundance in 2012. There were no correlations between water temperature or water transparency and biomass/abundance of P. magnifica during the growing season. P. magnifica colonies preferred to grow on the branches or roots (especially of Salix sp.) to aquatic macrophytes and stones. Most of the water bodies, where this bryozoan species occurred, had lower concentration of total phosphorus in the water when compared with the typical fishponds in the Czech Republic. Inner space of colonies of P. magnifica provided suitable higher trophic level substrate when compared with the water of the sandpits/fishponds especially for green coccal algae. A massive algal colonization was indentified in decomposing colonies at the end of the growing season.

Three species of Plectus Bastian, 1865 viz., P. aquatilis and P. pusillus from Kerman province and P. velox from Alborz province, Iran are described and illustrated. Partial sequences of 18S region of ribosomal DNA gene were amplified for P. aquatilis and P. pusillus. The Blast results of population of P. aquatilis from Iran showed 8–10 nucleotides differences with populations of the same species (AF036602; GQ892827; AY284700) reported from the UK, Belgium and The Netherlands, respectively. Whereas Iranian population of P. pusillus showed 14–16 nucleotides differences with P. cf. pusillus (AY284705; AY284704) reported from The Netherlands. Molecular analysis revealed close relationship of the Iranian plectids with P. cf. parvus (AY284699) reported from The Netherlands. Phylogenetic relationships with other related species in the genus Plectus and closely related genera that are available in the GenBank are given.

The main goals of this study were to determine the richness and diversity of helminth parasites of Rhinella fernandezae at the component and infracommunity levels and determine the ecological implications of different biotic and abiotic factors. Specimens were collected near the city of Corrientes, Corrientes Province, Argentina. Prevalence of infection was 94% in the specimens examined (n = 65). The helminth component community in R. fernandezae in this area was comprised a total of 22 species. Of all helminth species, only three (Catadiscus inopinatus, Cosmocerca podicipinus and C. parva) were dominant (importance value: I > 1.0) in the community. The most abundant species were B. tetracotyloides (d = 0.43) among the larvae and C. podicipinus (d = 0.09) among adult worms. At the infracommunity level, the mean individual species richness (2.28 ± 1.48) (mean ± SD) was no more than 3 helminth species per infected host; the diversity and equitability of helminths were 0.23 ± 0.21 and 0.48 ± 0.38, respectively. The host body size was the main factor in determining the parasite abundance. Species richness was significantly and positively correlated with host body size. The parasite helminth species predominantly showed an overdispersed pattern of distribution. Helminth species showed two negative and significant pairs of covariation and one significant pair of association (P < 0.05). R. fernandezae has a wide variety of parasites relating to the host microhabitat, mobility and feeding habits.

The diversity and longitudinal variation of zooplankton in the lower Vistula River were analyzed. Samples were taken from 40 stations located along a 272-km long section of the lower river course. During the study the unique technique of taking samples from “the same water” was used. The zooplankton community was dominated by rotifers and nauplii — larval stages of copepods. The most abundant species were: Brachionus angularis, Brachionus calyciflorus and Brachionus budapestiensis. The zooplankton species diversity in the main channel of the lower Vistula River was similar to other large European rivers; however, its abundance was lower. The diversity, abundance and biomass of potamoplankton steadily decreased downstream. This could be related both to scarcity of storage zones for potamoplankton development in the river due to the extensive regulation processes, and changes in hydrological conditions of the main channel (by the straightening of riverbed) where the samples were collected.

Three new oribatid mite species, Belbodamaeus indicus sp. n. (Damaeidae), Malaconothrus macrofoveolatus sp. n. (Malaconothridae) and Nothrus phylliformis sp. n. (Nothridae), are described from Indian soils. Belbodamaeus indicus sp. n. is clearly distinguishable from all species of Belbodamaeus by the absence of discidia, very long sensilli and morphology of parastigmatic tubercles Sa. Malaconothrus macrofoveolatus sp. n. is very similar morphologically to Malaconothrus yinae Yamamoto, Aoki, Wang & Hu, 1993 from China, however it differs from the latter by the morphology of notogastral and genital setae, size of body foveolae, epimeral formula, and number of genital setae. Nothrus phylliformis sp. n. is very similar morphologically to Nothrus mystax Mahunka, 1986 from Tanzania, however it differs from the latter by the smaller body size, length of interlamellar setae and the position of notogastral setae d 1. An identification key to known species of Belbodamaeus is presented.

We report on occurrences of the subcosmopolitan palpigrade Eukoenenia florenciae (Rucker, 1903) in a hothouse in Bratislava and in a dry river bed on Tenerife. We redescribe this probably parthenogenetic species, provide information on the thick setae of the coxae and the forked setae on the terminal segment of the first leg, and discuss the significance of these previously disregarded characters for species delimitation and systematics of the Palpigradi.

Descriptions of Cerapachys browni sp. n. and Cerapachys costatus sp. n. from India are provided. C. browni is the member of the wide spread dohertyi group while C. costatus represents the 3rd species of the rare crawleyi group, with C. sauteri Forel, 1913 and C. crawleyi Wheeler, 1924 reported earlier. An identification key to the worker caste of Indian Cerapachys species is provided.

We studied differences in respiration of materials from different parts of wood ant nest (top, bottom, and rim) and from the nest surroundings (humus layer and mineral soil). Samples were taken from 8 wood ant (Formica aquilonia) nests in each of the two types of forest (birch and pine) in eastern Finland. The differences were related to material and forest stand characteristics (i.e., moisture, pH, carbon content, and C:N ratio). As a result, the highest respiration per g DW was measured at the top of ant nests in the birch forest. However, respiration did not significantly differ between the parts of ant nests in the pine forest. Respiration of the humus layers in both forest stands was on average higher, whereas respiration of the mineral soils in both forest stands was lower in comparison with respiration of the nest materials. The respiration per g C did not show any significant differences between different parts of nests and surrounding soil. The most important factors influencing respiration of the materials appeared to be moisture, carbon content, and pH. In conclusion, respiration of wood ant nest material is affected by the specific material and forest stand characteristics.

Succession of Carabidae communities in spruce forests in the High Tatra Mts damaged by the windstorm of November 2004 exhibited two trends. The first trend includes the communities differentiation according to the state and management of damaged sites into three groups: (1) the site with fallen timber in situ shows only quantitative and reversible changes in rapport to the intact stand, (2) the sites with extracted timber, where less tolerant forest species disappeared, more tolerant forest species were favored and non-forest mountain species appeared, (3) the sites with extracted timber, additionally burned in July/August 2005, where number of the forest species and their abundance declined and temporal invasions of xenocoenous open-landscape species occurred. This differentiation is explained by autecology of individual species and state of vegetation. In 2010, the communities in burned and unburned sites started to converge due to partial restoration of the vegetation cover, but they continued to strongly differ from the site with timber in situ. The second trend includes a striking decline of the number of species and individuals and cumulative biomass in all sites in 2008 and a slow increase of these parameters up to 2011. The extreme dry summer of 2012 caused a decline of these parameters about to the levels from 2009. This trend represents a long lasting consequence of the extremely dry year 2007 and an immediate response to the drought in 2012. The Standardized Precipitation Index (SPI24) and Standardized Evapotranspiration Index (SPEI24) fitted best these changes.

The primary aim of this study was to identify reference genes and workers of particular role and ages that would be suitable for exploring genetic/epigenetic variations in constitutive expression of a gene encoding antimicrobial peptide defensin1 in worker heads using real-time PCR. This peptide is an integral component of larval food and honey and has potential to act against some brood pathogens. Expression levels of distinct genes may vary in worker heads due to genetic factors, age of bee, and particular role of a worker that depends on its age or colony needs. Prerequisite for exploring the variations in defensin1 expression was therefore to identify such workers in which correlated expression of defensin1 and suitable reference genes occurs. Selection process was done by carefully designed quantitative real-time PCR procedure in two colonies showing different age-related division of labor. Expression of ten candidate reference genes, defensin1 and amylase, as a marker of forager bees, was assessed in pooled head samples of workers aged 2 to 30 days. Correlated and moreover stable expression of defensin1 and six candidate genes was detected in nursing bees in both colonies. The suitable reference genes were therefore selected on the basis of their expression stability. This was evaluated by geNorm and NormFinder algorithms in pooled head samples and through plotted Cq data in head samples of individual nurse bees. As the best reference genes were selected: psa1, tctp1, cyclophilin, gapdh and mrjp4 (in this order). They are suitable for aforementioned defensin1 expression studies and also for studies of other genes expressed in heads of nurses. In addition, an amylase expression-based procedure for reliable distinguishing nurses from foragers was elaborated.

A genetic linkage map of Japanese flounder was constructed using 165 doubled haploids (DHs) derived from a single female. A total of 574 genomic microsatellites (type II SSRs) and expressed sequence tag (EST)-derived markers (EST-SSRs) were mapped to 24 linkage groups. The length of linkage map was estimated as 1270.9 centiMorgans (cM), with an average distance between markers of 2.2 cM. The EST-SSRs were used together with type II SSR markers to construct the Japanese flounder genetic linkage map which will facilitate identify quantitative trait locus (QTL) controlling important economic traits in Japanese flounder. Thus, twelve skeletal traits at 2 years of age were measured for all DHs. Forty-one QTLs were detected on 14 linkage groups and totally account for a small proportion of phenotypic variation (4.5 to 17.3%). Most of QTLs detected distribute on linkage groups 5 (9 QTLs), 8 (9 QTLs), 9 (5 QTLs) and 20 (4 QTLs), in which, some QTLs perform the pleiotropy.

The study focuses on the phylogenetic origin and genetic diversity of brown trout in the eastern part of the Balkan Peninsula. It further aims to reveal the impact of human-mediated transfers and stocking with non-indigenous trout on the populations in this area. For these purposes, mtDNA control region and microsatellite variation of 204 individuals from 16 populations were analysed. The results indicate that mtDNA haplotypes from the lower Danube basin and southern Black Sea basins differ substantially from a subclade of the Danubian lineage consisting of haplotypes found so far in the most of the Danube basin and in the Caspian and Aral Sea basins. Considering also the results of demographic analyses, this study evidences a complex evolutionary history of brown trout in the southern and western parts of the Black Sea basin. In the Aegean Sea basin, a high frequency of the central haplotype of Adriatic mtDNA lineage has been found. The other Adriatic lineage haplotypes found in this basin differ from the central haplotype by one mutational step only, indicating a recent evolution of the Adriatic lineage in the Aegean Sea basin. Substantial genetic differentiation among populations and basins was revealed. The hybridization with Atlantic brown trout was indicated in both sea basins, but especially in the Danube basin. Compared to other European regions, it can be inferred that the introgression of exogenous brown trout in the eastern Balkan populations is rather low.

The present histochemical study was carried out to analyze the distribution and topography of acetylcholinepositive nerve fibers in the thymus and bursa of Fabricius of quails. The AChE-positive nerve fibers were demonstrated by direct thiocholine histochemical method. Nerve fibers present in the thymuses form periarterial nerve plexuses located mostly in the interlobular septa and on the cortico-medullary junction. Vessels-independent nerve fibers occur also in the parenchyma of thymic medulla, but rarely in parenchyma of the cortex. Within the connective tissue between the bursa of Fabricius and the wall of proctodeum we observed conspicuous AChE-positive ganglia, often in close relationship to greater arteries. Within the wall of bursa of Fabricius, AChE-positive nerve fibers create nerve plexuses around arteries. We observed a close relationship between lymphoid follicules in bursal submucosa and mucosa and AChE-positive nerve fibers. Nerve fibers create a ring around lymphoid follicles, but do not penetrate into the germinal center of the follicle. Arteries inside quail thymuses and bursae of Fabricius contain rich AChE-positive nerve plexuses, when compared to the veins, which have a very poor presence of AChE-positive nerves. According to lesser presence and decreased density of AChE-positive nerve fibers in older animals, we described age-dependent changes in both quail primary lymphoid organs.