Volume 66, Issue 2

Heavy metal contamination of soil, aqueous waste stream and ground water causes major environmental and human health problems. Heavy metals are major environmental pollutants when they are present in high concentration in soil and show potential toxic effects on growth and development in plants. Due to unabated, indiscriminate and uncontrolled discharge of hazardous chemicals including heavy metals into the environment, plant continuously have to face various environmental constraints. In plants, seed germination is the first exchange interface with the surrounding medium and has been considered as highly sensitive to environmental changes. One of the crucial events during seed germination entails mobilization of seed reserves which is indispensable for the growth of embryonic axis. But, metabolic alterations by heavy metal exposure are known to depress the mobilization and utilization of reserve food by affecting the activity of hydrolytic enzymes. Some plants possess a range of potential mechanisms that may be involved in the detoxification of heavy metals by which they manage to survive under metal stress. High tolerance to heavy metal toxicity could rely either on reduced uptake or increase planned internal sequestration which is manifested by an interaction between a genotype and its environment. Such mechanism involves the binding of heavy metals to cell wall, immobilization, exclusion of the plasma membrane, efflux of these toxic metal ions, reduction of heavy metal transport, compartmentalization and metal chelation by tonoplast located transporters and expression of more general stress response mechanisms such as stress proteins. It is important to understand the toxicity response of plant to heavy metals so that we can utilize appropriate plant species in the rehabilitation of contaminated areas. Therefore, in the present review attempts have been made to evaluate the effects of increasing level of heavy metal in soils on the key behavior of hydrolytic and nitrogen assimilation enzymes. Additionally, it also provides a broad overview of the strategies adopted by plants against heavy metal stress.

In recent years, α-galactosidase has been attracting more and more attention because of its potential applications in many aspects. Using reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends, a full-length cDNA sequence composed of 2,439 bp was cloned from Penicillium janczewskii zaleski and was subcloned into pPICZαA and transformed into Pichia pastoris strain X-33. In a 10-L fermentor, the recombinant yeast expressed α-galactosidase with a yield of 254 U/mL by methanol induction for 120 h. The recombinant enzyme showed the optimal activity at 40°C and pH 5.2. The K m values of the recombinant enzyme using p-nitrophenyl-α-D-galactopyranoside (pNPG), melibiose, raffinose and stachyose as substrates were 1, 16, 17.8 and 5.3 mM, respectively. V max values were 227.3, 116.7, 104.8, and 80.6 μM/min using pNPG, melibiose, raffinose and stachyose as substrates, respectively. The α-galactosidase exhibited no sensitivity to various metal ions and ethylenediaminetetraacetic acid, and hydrolyzed melibiose, raffinose and stachyose with different levels of galactose release. The biochemical characteristics of the α-galactosidase suggest that the enzyme may have a prospective application in feed industry as an additive.

A 1.4 Kb fragment of Bacillus licheniformis ATCC 14580 encoding β-glucosidase was cloned and expressed in Escherichia coli. β-Glucosidase expressed by E. coli harboring cloned gene was located entirely in the intracellular fraction. Recombinant β-glucosidase protein was purified to homogeneity level and the molecular weight was found to be 53 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. It gave maximum activity at 50°C and pH 6. K m and V max were 0.206 mM and 1.26 U/mg, respectively, with p-nitrophenyl-β-D-glucopyranoside, while activation energy Ea, enthalpy of activation ?H and entropy of activation ΔS were found to be 66.31 kJ/mol, 64.04 kJ/mol and 48.28 J/mol/K, respectively. The pKa1 and pKa2 of the ionizable groups of active site residues involved in Vmax were found to be 5.5 and 7.0, respectively. When the recombinant β-glucosidase protein was used as a member of consortium with endoglucanase and exoglucanase for the saccharification of wheat straw, 123% increase in saccharification was observed.

Complete nucleotide sequence of plasmid pGP2 from Acetobacter estunensis GP2 was identified after initial cloning of EcoRI fragment followed by preparation of deletion derivatives. Its size was defined to 2,797 bp and several sites for several restriction enzymes were revealed by DNA sequencing. Sequence analysis predicts three putative open reading frames (ORFs). ORF1 shows significant identity with the bacterial excinuclease α-subunit, ORF2 is a putative replication protein with low similarity with other Acetobacter plasmid’s replication proteins, and ORF3 encodes a class B acid phosphatase/phosphotransferase. The replication module comprises a DnaA box like sequence, direct repeats, a potential prokaryotic promoter and a rep gene. The rep module is similar with several θ-replicating, iteron-containing modules from plasmids, suggesting pGP2 replication may follow the same course. Any phenotypic character determinant gene is absent in pGP2, suggesting this plasmid to be cryptic. However, a pGP2 derivative plasmid, containing the putative pGP2 rep region, can replicate and is stably maintained in Acetobacter and Escherichia coli strains; it can also carry foreign DNA fragments. Thus, pGP2-X could serve as a cloning shuttle vector between these bacteria. Prepared deletion derivatives of plasmid pGP2 suggested that Rep protein is essential for plasmid replication in host bacteria. In its natural host, A. estunensis GP2, pGP2 maintains a four-times lower copy number than in E. coli.

Male and female gametophytes have special characters that show a great variety in different taxa. In this study, gametophytes of four species belonging to three sections of the genus Onobrychis Mill. were studied with light microscopy. Results showed that the ovular primordium is tetra-zonate and gives rise to an anatropous ovule. The archesporium may consist of one or more archeosporial cells, but only one of them undergoes meiosis, forming a linear or T-shaped tetrad. Normally, only a single megaspore is functional which is located in the chalazal position while the others degenerate very soon. The young ovule is hemi-anatropous but the mature is anatropous, crassinucellar and bitegmic; integuments form a zig-zag micropyle. A 7-celled embryo-sac is formed corresponding to the Polygonum type. Based on our results, the ovular variable characters are the form and condition of ovary, presence or absence of ovary peduncle, the number and condition of ovule in ovary, length and width of ovule, length and width of embryo sac, number of layers in outer integument, condition of megaspore, alignment pattern of the integuments, asymmetrical initiation of the outer integument, shape of tetrad with the presence of one functional megaspore and so on. The separator characters in male gametophyte are including tri-cellular pollen grains and the number of tapetum nuclei. According to our study the female gametophyte characters are more variable than male gametophyte. The present study provides the first report on embryological description in the genus Onobrychis and also in section Heliobrychis.

PCR amplification techniques viz., repetitive DNA element PCR (REP-PCR), short tandemly repeated repetitive PCR (STRR-PCR) and arbitrarily primed PCR (RAPD-PCR) were used for the taxonomic discrimination among the strains of the unicellular cyanobacterium Synechococcus elongatus collected across the coastal regions of the Indian subcontinent. These strains showed similar phenotypic and genotypic characteristics. Data obtained from genomic fingerprinting were used to perform cluster analysis and demonstrated ability to differentiate strains at intra-specific level. Polymorphisms of different PCR amplification products can serve as strain-specific molecular fingerprints. In comparison with the STRR and RAPD, the REP primer set generates fingerprints of lower complexity, but still the phenogram clearly differentiated the strains. In conclusion, described PCR fingerprinting methods can be considered as promising tools for the differentiation at the strain level of cyanobacteria from the same species.

Conservation of identified germplasm is an important component for efficient and effective management of plant genetic resources. Since Chlorophytum species are important medicinal plants, studies were carried out for identification and establish genetic relationships in three species of Chlorophytum and two high yielding genotypes of Chlorophtum borivilianum using RAPD markers. Out of one hundred primers tested, 47 decamers amplified a total of 454 distinct bands ranging from 0.25–3.0 kbp to identify and to evaluate genetic relationships between and among three species of Chlorophytum and two genotypes of Chlorophtum borivilianum. The cluster analysis indicated that three species of Chlorophytum and two genotypes (NRCCB-1 and NRCCB-2) of C. borivilianum formed two major clusters. The first major cluster constituted C. arundinaceum and C. tuberosum, and the second major cluster composed of two subclusters; the first subcluster represented NRCB-1 and NRCB-2 where as the second subcluster represented C. borivilianum. Thus, the RAPD markers have the potential for identification and characterization of genetic relatedness among the species and genotypes. C. borivilianum along with two genotypes also showed similar banding patterns which could be chosen as candidate markers for differentiating the other two species such as C. arundinaceum and C. tuberosum. This would helpful for breeding programmes and provides an important input in conservation biology.

KNOX (KNOTTED1-like homeobox) genes encode homeodomain-containing transcription factors which play crucial roles in meristem maintenance and proper patterning of organ initiation. PttKN1 gene, isolated from the vascular cambium of hybrid aspen (Populus tremula × P. tremuloides), is a member of class I KNOX gene family. In order to understand the roles of PttKN1 gene in meristem activity and morphogenesis as well as to explore the possibility to generate novel ornamental lines via its ectopic expression, it was introduced into the genome of Begonia maculata Raddi by Agrobacterium tumefasciens-mediated gene transformation here. Four types of transgenic plants were observed, namely coral-like (CL) type, ectopic foliole (EF) type, phyllotaxy-irregular (IP) type and cup-shaped (CS) type, which were remarkably different from corresponding wild type and were not also observed in the regenerated plantlets of wild type plant. Among these four types of transgenic plants, the phenotype of coral-like was observed for the first time in the transformants ectopically expressed KNOX genes. The observation of scanning electron microscope (SEM) showed ectopic meristems on the adaxial leaf surface of the transformants. Interestingly, the plantlets with ectopic foliole could generate new ectopic folioles from the original ectopic folioles again, and the plants regenerated from the EF-type transformants could also maintain the original morphology. The same specific RT-PCR band of the four types of transgenic plantlets showed that PttKN1 was ectopically expressed. All these data demonstrated that the ectopic expression of PttKN1 caused a series of alterations in morphology which provided possibilities producing novel ornamental lines and that PttKN1 played important roles in meristem initiation, maintenance and organogenesis events as other class I KNOX genes.

Serpentine outcrops are distinct from their background geological context, as they are often isolated from one another, and provide a home to a significant number of rare or endemic taxa. The aim of this study was to assess the Portuguese populations of one such taxon, Notholaena marantae subsp. marantae, including information on its biogeography, habitat, geographical range and conservation status. A detailed study was also made of the endemic association Notholaenetum marantae, a member of the Phagnalo saxatilis-Cheilanthion maderensis alliance, a type of vegetation included in the EU Habitats Directive 92/43/ECC. The distribution of Notholaena marantae was highly fragmented, with two sites accounting for over 50% of the total number of individuals recorded. The presence of N. marantae is associated with that of ultramafic rocks, which show specific geochemical features. The conservation status of the species is a cause for concern. We propose that the species be considered ‘vulnerable’ according to the criteria of the IUCN (2001), and that areas where its populations grow be protected. This study confirms the endemic status of N. marantae subsp. marantae in Portugal and the importance of the conservation of its serpentine habitats.

The flora of the White Carpathians, a mountain range in the south-east of the Czech Republic, is documented by about 485,000 records of vascular plant occurrences collected since the mid-19th century. A total of 1299 species recorded in 93 grid cells of 2.8 × 3.1 km were used for an analysis of spatial patterns of floristic diversity in the White Carpathians. Multivariate statistical techniques such as ordination and classification were used to reveal the main gradients in floristic composition and species richness, and measured environmental data and Ellenberg indicator values were used to assess underlying environmental factors. There is a striking floristic contrast between the western and eastern part of the study area, which is associated with differences in climate, mean altitude, topographic heterogeneity measured as altitudinal range, and land use. The western part is characterised by thermophilous, continental and calcicolous species of open habitats. In contrast, the more forested eastern part along the state border with Slovakia and the north-eastern part of the area are characterised by acidophilous species with higher moisture requirements. This pattern is consistent with the established phytogeographical division of the Czech Republic into the phytogeographical regions of Thermophyticum and Mesophyticum. The further division of the area into four regions, based on classified grid data, is also similar to the current division into phytogeographical districts, except for the Javorníky district. There are two distinct hot spots of species richness, in the western and the extreme north-eastern part. A poorer flora was found in landscapes with intensive agriculture. Species richness is associated with different environmental factors than species composition, namely with soil types and land-use categories. Alien species are more common in areas with a higher incidence of arable land and built-up areas, and less common in areas dominated by grasslands and forests.

Tall expansive grasses act as serious weeds since they spread intensively and are a important threat to biodiversity of various plant communities. A field experiment was set up based on three sets of paired plots, where Calamagrostis epigejos and Arrhenatherum elatius dominated and a mixture of both these tall grasses occurred. Parallel plots were treated by additional amounts of nitrogen (50 kg N ha−1) for six years (2002–2007). Above-ground biomass was harvested in both wet (2004) and dry (2007) years, separated in individual fractions, and weighed to determine the above-ground biomass. We wanted to know if a higher nitrogen availability in dry year can support above-ground growth of expansive tall grasses C. epigejos and A. elatius and what their competitive ability of these tall grasses in mixture is in dry conditions. Drought resulted in a decrease in above-ground biomass production of studied tall expansive grasses, and, in a relative increase in mortality of A. elatius shoots in comparison with C. epigejos. Drought can substantially reduce the effect of applied nitrogen on above-ground growth of expansive tall grasses. However, A. elatius appears to be producing more above-ground biomass after application of N even in relatively dry conditions.

Prevalence of the infectious respiratory agens, avian influenza virus (AIV), Mycobacterium avium (M. avium), and Mycobacterium avium subspecies paratuberculosis (MAP), was studied in migratory marsh-dwelling passerines captured in the Parížske močiare wetlands in Western Slovakia during 2008. Surveillance of 650 birds revealed a lower prevalence of AIV in spring (13.6%) than in summer (17.5%). A total of 14 different subtypes were detected in samples obtained from birds captured during the spring, with the most prevalent subtypes being H8N3, H6N4, H11N6 and H12N6. Subtypes H12N6, H6N6 and H2N5 were predominant in passerines captured during summer months. In eight cases, different AIV infections were detected in the oropharyngeal and cloacal samples originating from a single bird (H1N1 and H8N3; H1N3 and H9N3; H2N3 and H12N6; H2N1 and H8N1; H4N2 and H9N6; H5N5 and H11N6; H6N4 and H11N6; H7N1 and H10N3 in the oropharynx and cloaca, respectively). M. avium was detected in 9.2% and 0.8% of marsh-dwelling passerines captured during spring and summer, respectively. Only two birds were co-infected with AIV and M. avium. All birds were negative for MAP.

Colorado potato beetle, Leptinotarsa decemlineata (Say), is a devastating pest of potatoes in North America and Europe. L. decemlineata has developed resistance to insecticides used for its control. In this study, in order to find a more effective potential biological control agent against L. decemlineata, we investigated its microbiota and tested their insecticidal effects. According to morphological, physiological and biochemical tests as well as 16S rDNA sequences, microbiota was identified as Leclercia adecarboxylata (Ld1), Acinetobacter sp. (Ld2), Acinetobacter sp. (Ld3), Pseudomonas putida (Ld4), Acinetobacter sp. (Ld5) and Acinetobacter haemolyticus (Ld6). The insecticidal activities of isolates at 1.8×109 bacteria/mL dose within five days were 100%, 100%, 35%, 100%, 47% and 100%, respectively, against the L. decemlineata larvae. The results indicate that Leclercia adecarboxylata (Ld1) and Pseudomonas putida (Ld4) isolates may be valuable potential biological control agents for biological control of L. decemlineata.

This study presents the parasitical relationships between the leech Piscicola respirans and European grayling Thymallus thymallus. It determines leech invasion intensity and extensity as well as describes the preferences of fin selection by the parasite as a place to attach and feed. It also shows the significance of fin selection by the leech in the initial phase of life cycle. Finally, the study defines P. respirans as a predominantly fin-parasite of fishes. Most probably the host-searching mechanism is based on positive phototaxis.

This paper presents a part of the results obtained from extensive studies conducted on springs — remarkable freshwater habitats with many unique characteristics and a high degree of biological diversity. The abiotic conditions and fauna composition of 68 submontane permanent oligotrophic springs in the Blanice River catchment (eastern part of The Bohemian Forest, Czech Republic) were investigated with a focus on their relationship to the Pisidium species occurrence. We attempted to determine the habitat requirements of individual pea clam species. Species of genus Pisidium generally preferred unshaded springs with fine substrate, the most favourable environment were helocrene springs. Although they were the second most numerous taxonomical group in the springs, only two species were present there: Pisidium casertanum and P. personatum. The occurrence of both species strongly positively correlated with each other. Substrate type and a few chemical parameters were the main parameters, which differentiated the distribution of both species. While P. casertanum was found to be most abundant in vegetation, P. personatum preferred humolite substrate. The occurrence of P. casertanum was further influenced by nutrients compared to P. personatum, whose occurrence positively correlated with the common minerals. In general, biotope characteristics of springs were found to be more important for Pisidium species inhabiting oligotrophic mountain springs than water chemistry.

The investigation of the vertical distribution of the zooplankton community in the temperate Lake Trichonis during four seasons in 2005, showed the existence of vertical segregation among species, ontogenetic stages and sexes within and between the major groups. In each season, the two or three more abundant rotifer species distributed at separate depth layers, while this feature was maintained during the entire 24 h period, since no diel vertical migrations (DVM) were performed. In contrast, the crustacean community, comprised mainly by the calanoid copepod Eudiaptomus drieschi and the cladoceran Diaphanosoma orghidani, showed various patterns of DVM, being more pronounced in spring and summer. Females of E. drieschi distributed deeper than males, while the copepod nauplii were found mainly in the surface layer in all four seasons. Temperature was the most important abiotic factor affecting directly and indirectly the vertical distribution and migration of various species. During stratification, the metalimnion was the most productive layer in Lake Trichonis, having maximum values of dissolved oxygen and low transparency due to high concentration of organic matter and phytoplankton. The DVM patterns of the crustaceans indicate that the metalimnion acts probably as a daylight refuge against predation by Atherina boyeri, which is the dominant planktivorous fish in the lake.

The historical influence of landscape on the quantitative variables and a special group of hyporheos in streams with similar origin but different catchment landscapes and forest management was studied. The study was conducted in two streams: (i) a preserved forested natural stream where critical ecosystem processes were unaltered by human activities and (ii) a stream with the strong anthropogenic stressors (e.g., logging, agriculture, pasture), impacting the system for the last 500 years. Some parameters were strongly related to these land use: conductivity, fine benthic organic matter (FBOM), orthophosphate (P_PO4) and periphyton content all increased along the gradient from the natural stream to the urban-dominated catchment. The density of interstitial assemblages corresponded with the conductivity (at P < 0.05) and was higher on the stream in urban-dominated catchments; and the Harpacticoida taxa richness, dominated by the family Canthocamptidae, was also greater here. The Multiple Regression Analysis shows that the rate of deforestation had had the most significant effect on the density of hyporheos. Among of crustacean fauna the representatives of epigean harpacticoids from (of) family Canthocamptidae, dominated.

Correspondence of habitat and macroinvertebrate diversity was investigated in twenty Czech rivers. With regard to the importance of river characteristics (depth, current velocity and substratum) River Channel Habitat Diversity method was developed within this paper. The method is focused directly on the stream wetted channel and it assesses habitat diversity and richness computed as Shannon-Wiener’s index. Macroinvertebrate samples were taken by the national PERLA method at each study site, diversity index was also calculated and compared with habitat and substrate diversity. Habitat diversity was important for macroinvertebrate communities according to multivariate assessment, but contrary to the hypothesis it was not significantly correlated with macroinvertebrate diversity. Based on the results, broader rivers had lower habitat diversity than smaller streams, and were more influenced by human activities (lower habitat diversity, higher pollution). Results obtained by the RCHD and RHS methods were not significantly correlated.

We examined the species composition and community structure of mites of the order Mesostigmata (Acari) in nests of the Eurasian griffon vulture (Gyps fulvus Hablizl, 1783) in Croatia. Material collected from 18 nests included 565 mites belonging to seven species. The most abundant species were Leiodinychus orbicularis (C.L. Koch, 1839) (Trematuridae) and Androlaelaps casalis (Berlese, 1887) (Laelapidae). The results were compared with the community structure and frequency of dominant species of Mesostigmata in nests of 32 other bird species. Leiodinychus orbicularis occurred in the nests of 13 species of birds. It is a typical nidicolous species which occurs most frequently in the perennial nests of birds of prey. In contrast, A. casalis rarely occurs in the nests of birds of prey.

To determine the cotton bollworm migrating population rate in Hungary, we examined the weights and the front wing morphological feautures of trapped moths. We used sex pheromone traps to monitor field populations during the maize vegetation cycle period in 2008. We examined moths trapped at various times, and measured their body mass (m) and morphological features, namely the front wing quotient (fWQ = quotient of length of front wing/width of front wing), modified wing loading (WL = weight of moth/surface of front wing), and the relative thorax size (RTS = width of thorax/width of head). The data were analysed by Student t-test, anterior wing abrasion and darkness were analysed by a Adobe Photoshop 7.0 software. The Hungarian appearance of three cottom bollworm generations in 2008 was also observed. Based on the examined morphological features we found regularity in body mass, front wing quotient and modified wing loading changes during the flight period. The specimens trapped in the first and third part of the flight period had lower body mass, larger wing surface, longer wings and more favourable modified wing loading than the specimens trapped in the middle of the flight period. The abrasion and colour of the anterior wings of cotton bollworms were concordant to morphometric investigations. The abrasion in darker spots E1 and E3 clearly showed a more intensive usage of the wings in case of specimens trapped at the beginning and at the end of the flight period.

In this study, for the first time, the morphology of the dorsal and ventral skin of Triturus karelinii using light microscopy through histochemical methods was described. The skin exhibited basic morphological characteristics of the other urodeles: the epidermis composed of keratinized stratified epithelium with numerous conical protrusions and the dermis subdivided into spongy and compact layers. In the spongious dermis, three distinct types of glands were observed, namely serous, mucous and mixed glands. These glands were alveolar and occurred in both males and females. The morphologies of all three skin glands differed from anurans and other urodeles, having peculiar characteristics. Serous glands exhibited three different appearances. An unusual finding in mucous glands was the different appearance of their granules, showing diverse density after staining with PAS and AB. The histochemical analysis clearly demonstrated the presence of neutral, acid and sulfated mucins in the adenocytes of mucous glands. Mixed glands formed by mucous and serous glands exhibited the same morphological traits of both types of glands. Light microscopic observations revealed that the dorsal and ventral skin showed structural similarities with some minor differences, possibly resulting from their functions.

Selenium is an essential element with a narrow margin between beneficial and toxic effects. This study was aimed to determine the neurobehavioral changes resulted from the prenatal exposure of mice to high doses of sodium selenite during fetal and early postnatal development. Atomic absorption for monitoring the placental transfer of selenium to offspring was employed. The developmental observations as well as the behavioral tests, such as sensory motor reflexes, and learning and memory test in automatic reflex conditioner (shuttle box) (active avoidance responses) were applied. Adult mice was assigned into three groups: the first group was remained as a control group given phosphate buffered saline; the second and the third groups were orally administrated sodium selenite at doses of 1 mg/kg and 4 mg/kg of the diet, respectively started from the 7th day to the end of the gestation period. Appearance of body hair and opening of eyes of the pups from treated mothers were delayed in a dose-dependent manner. The body weight gain came significantly lower than those of the control especially at the higher dose. Selenite also inhibited the sensory motor reflexes in all elements of acts and postures in a dose dependent manner. The active avoidance training-test indicated that selenite exposure was associated with learning impairment. Acetylcholine recorded a significant decrease in almost all the period of this study. By using atomic absorption, we found a significant high concentration of selenium in the brain, liver and kidney until the 40th postnatal day, indicating active transfer of selenium from mothers to embryos.

The effects of retinoic acid analogue — phytol (precursor of retinoic X receptors ligand), TTNPB (retinoic acid receptor agonist) and seocalcitol (EB1089, analogue of vitamin D3) in mammary tumours of Sprague-Dawley rats induced with 1-methyl-1-nitrosourea (MNU) were investigated. Treatment with phytol, TTNPB and seocalcitol may have some protective and therapeutical effects on malignant processes. Treatment with these components in combination of TTNPB and phytol or seocalcitol and phytol inhibited progression of MNU-induced tumours of the rat mammary gland, and also induced decrease of tumour burden and volume in comparison with treated control group. Treatment of rats with the above compounds had no effect on malignity and invasiveness of carcinomas.

The preorbital gland is a specialized dermal gland of antelopes which plays an important role in territorial marking behavior and pheromonal communication. To our knowledge, there is little information available on the role of preorbital gland marks in Indian antelopes (blackbucks). Males are seen averting the gland during behavioural display and territorial marking but the functional aspect of this gland has not been examined. Hence, the aim of this study was to describe the histomorphology of the preorbital gland in territorial and non-territorial male blackbucks to determine its morphology and secretory function. The results showed that the preorbital gland is composed of modified sebaceous and apocrine glands. The apocrine gland is lined by simple cuboidal epithelial cells; the serous parts of the secretory products are often seen in the apical portions of the cells. The myoepithelial cells contain actin filaments lying on the basal membranes of the apocrine gland. There are some considerable histological changes in the presence of the sebaceous and apocrine glands in territorial males in comparison to non-territorial males. The following histological changes associated with occurrence of the sebaceous and apocrine glands have been observed in territorial and non-territorial male blackbucks: (1) increase of size of sebaceous and apocrine glands and (2) increase in density of sebaceous and apocrine glands in territorial males compared to non-territorial males. It is suggested that the higher development (i.e., size) and density of sebaceous and apocrine glands in territorial males could depend on hormone production (i.e., testosterone). Based on the histological observation and the role of sebaceous and apocrine glands in the preorbital gland supported by literature, it is possible to conclude that both territorial and non-territorial blackbuck males may produce pheromonal substances through preorbital gland (secretion) for olfactory communication.

The molecular mechanisms of phenoxyherbicides action in animals have been insufficiently studied. Now, we have investigated the interaction of sodium salts of phenoxyherbicides, e.g., 2,4-dichlorophenoxyacetic acid (2,4-D-Na), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T-Na) and 4-chloro-2-methylphenoxyacetic acid (MCPA-Na) with human erythrocytes. In this study, we evaluated the effect of these compounds on erythrocyte membrane fluidity as well as changes in membrane proteins content. It was observed that all of the compounds studied altered membrane fluidity, changed the size and shape of the erythrocytes and provoked echinocytes formation. It was also revealed that 2,4-D-Na and 2,4,5-T-Na changed the content of erythrocyte membrane proteins mainly by a decrease in the level of spectrin and low molecular weight proteins. The comparison of the action of phenoxyherbicides examined showed that 2,4,5-T-Na caused the greatest changes in the erythrocytes membrane, whereas MCPA-Na induced the lowest alterations in the incubated cells. It must be noted that changes of the investigated parameters were observed only at presence of significant concentrations of these compounds that may penetrate human organism only as a result of acute poisoning.