In the present study a new luminescent dye 3‐N‐(2‐pyrrolidinylacetamido)benzanthrone (AZR) was synthesized. Spectroscopic measurements of the novel benzanthrone 3‐aminoderivative were performed in seven organic solvents showing strong fluorescence. The capability of the prepared dye for visualization has been tested on flax, red clover and alfalfa to determinate the embryo in plant callus tissue cultures. Callus cells were stained with AZR and further analysed utilizing confocal laser scanning fluorescence microscopy. Performed experiments show high visualization effectiveness of newly synthesized fluorescent dye AZR that is efficient in fast and relatively inexpensive diagnostics of callus embryos that are problematic due to in vitro culture specificity.

Helminth infracommunities were studied at 174 sites of Latvia in seven hosts from six amphibian taxa of different taxonomical, ontogenic and ecological groups. They were described using a standard set of parasitological parameters, compared by ecological indices and linear discriminant analysis. Their species associations were identified by Kendall's rank correlation, but relationships with host size and waterbody area were analysed by zero-inflated Poisson and zero-inflated negative binomial regressions. The richest communities (25 species) were found in post-metamorphic semi-aquatic Pelophylax spp. frogs, which were dominated by trematode species of both adult and larval stages. Both larval and terrestrial hosts yielded depauperate trematode communities with accession of aquatic and soil-transmitted nematode species, respectively. Nematode loads peaked in terrestrial Bufo bufo. Helminth infracommunities suggested some differences in host microhabitat or food object selection not detected by their ecology studies. Associations were present in 96% of helminth species (on average, 7.3 associations per species) and dominated positive ones. Species richness and abundances, in most cases, were positively correlated with host size, which could be explained by increasing parasite intake rates over host ontogeny (trematode adult stages) or parasite accumulation (larval Alaria alata). Two larval diplostomid species (Strigea strigis, Tylodelphys excavata) had a negative relationship with host size, which could be caused by parasite-induced host mortality. The adult trematode abundances were higher in larger waterbodies, most likely due to their ecosystem richness, while higher larval abundances in smaller waterbodies could be caused by elevated infection rates under high host densities.

Benzanthrone derivates are now widely used in many industrial and scientific applications as dyes for polymers and textiles. In biochemical, biomedical and diagnostics investigations benzanthrone dyes are used as a lipophilic fluorescent probe since many benzanthrone derivates demonstrate bright fluorescence and they have ability to intercalate between membrane lipids. The aim of research presented here was to assess the luminescence ability of benzanthrone derivatives using microscopic visualization of biological objects. Accordingly, specimens of freshwater trematodes: Diplostomum spathaceum, Diplodiscus subclavatus and Prosotocus confusus, were stained by novel benzanthrone dyes using different fixatives. The samples were examined under a confocal laser scanning microscope. All of the dyes tested demonstrated good results for digestive and reproductive system visualization. Based on obtained results we conclude that benzanthrone dyes could be used for internal and external structure confocal laser scanning microscopic imaging of trematode specimens.

Luminescent derivatives of benzanthrone are becoming more useful based on their light-absorbing and fluorescent-emitting properties. Our previous studies showed that luminescent staining properties of the same benzanthrone dye differ for variable parasite samples. Therefore, two types of benzanthrone dyes were prepared. One has a strongly basic amidine group and a halogen atom, and the other has an amide moiety and a tertiary amine group. Trematoda Parafasciolopsis fasciolaemorpha is a liver fluke of a moose (Alces alces) and has a significant influence on the health and abundance of the moose population. Staining protocols for parasite P. fasciolaemorpha specific organ or organ systems imaging are mostly time-consuming and labor-intensive. The study aimed to compare the fixation technique and the staining protocol by synthesized benzanthrone luminescent dyes to determine detailed morphology, anatomical arrangement of the organ systems and gross organization of the muscle layers of P. fasciolaemorpha using confocal laser scanning microscopy. Luminophores were tested for samples fixed in different fixatives. Developed dyes and staining protocol resulting in imaging of all parts of trematode without additional sample preparation procedures, which usually are required for parasite examination. Obtained results confirmed that the most qualitative results could be reached using 3-N-(2-piperidinylacetamido)benzanthrone dye which has amide moiety and a tertiary amine group. Based on obtained results, 3-N-(2-piperidinylacetamido)benzanthrone gave more qualitative parasite visualization than 2-bromo-3-N-(N′,N′-dimethylformamidino)benzanthrone.

An increasing number of bacteria have been described as benefiting from interaction with free-living amoeba. The most common association between free-living amoeba and microorganisms is interaction of various non-pathogenic and pathogenic bacterial species with amoeba. Various pathogenic bacterial species have capacity to resist digestion by free-living amoeba, which has been observed by many researchers. Also, several of these pathogens are able to resist digestion by macrophages. In addition, free-living amoeba have been associated with several diseases in humans. Acanthamniioeba castella is an important predator of bacteria. It is a ubiquitous organism in water, soil, and air. Attention from a public health perspective is needed by investigation of interaction of foodborne pathogens and free-living amoeba. Bacteria can use free-living amoeba as reservoirs, mediators or vehicles, an infection route, “biological gym” and evolutionary crib or interaction may result in a close endosymbiotic relationship. The purpose of this review is to describe the interaction mechanisms between free-living amoeba and common bacteria species that survive in host cells.