GENETIC DIVERSITY AND POPULATION STRUCTURE OF THE EUROPEAN EEL (ANGUILLA ANGUILLA) IN BALTIC LAKELAND

Anguilla anguilla (Linnaeus, 1758) is the unique catadromous fish species in the Latvian fauna. The Baltic Lakeland area representing naturally recruited and introduced eels. In recent decades, its population has decreased not only in Latvia, but also in Europe. European eel critically endangered due to overfishing of glass eels, blocking of migratory paths, deaths in HPS turbines, water pollution and diseases and parasites. Only some bodies of water are freely accessible to natural migration of eels in Latvia.Currently, its position has been recognized as being critical, and a range of normative acts have been adopted for its restoration, such as the Regulation EC 1100/2007. The restocking programs of the European eel Anguilla anguilla have been conducted for nearly one century in Latvia.This study provides the first data on population structure of freshwater eels in Baltic Lakeland, for use in eel conservation and management of aquaculture on a regional and/or global scale. By analysing the sequences of the Cyt b gene of mtDNA for individuals caught in locations throughout in Baltic Lakeland, we determined the population genetic structure of A. anguilla in the area. The diversity of haplotypes was studied in ten waterbodies from part of Baltic Lakeland, namely Lake Sīvers, Lake Usmas, Lake Ķišezers, Lake Liepājas, Lake Alūksnes, Lake Rāznas, Lake Vialikija Švakšty, Lake Svir, Lake Myadzyel, Myadzelka river. Additionally, this study investigated the affinity of the Latvian populations to other A. anguilla populations around the world.This is the first report about eel’s population genetic diversity in Baltic Lakeland. Haplotype variation was different in all investigated waterbodies. In current studies seven new unique haplotypes were detected. Eel population in Baltic Lakeland shows quite high genetic diversity and rapid population expansions, which possibly is results of intensive restocking program. Sequences characterized Anguilla rostrata in Lake Alūksnes were detected

Identification of <i>Sarcocystis</i> and <i>Trichinella</i> Species in Muscles of Gray Wolf (<i>Canis lupus</i>) from Lithuania

Apicomplexan Sarcocystis and Trichinella nematodes are food-borne parasites whose life cycle is carried-out in various wildlife and domestic animals. The gray wolf (Canis lupus) is an apex predator acting as an ecosystem engineer. This study aimed to identify the species of Sarcocystis and Trichinella found in the muscles of gray wolves in Lithuania. During the 2017–2022 period, diaphragm, heart, and hind leg samples of 15 animals were examined. Microscopical analysis showed the presence of two types of Sarcocystis parasites in 26.7% of the analyzed muscle samples. Based on the sequencing of five loci, nuclear 18S rDNA, 28S rDNA, ITS1, mitochondrial cox1, and apicoplast rpoB, S. arctica, and S. svanai were identified. The current work presents the first report of S. svanai in gray wolf. Phylogenetically, S. svanai clustered together with S. lutrae, infecting various carnivorans, and S. arctica was most closely related to S. felis from domestic cats. Trichinella spp. were found in 12 gray wolves (80%). For the first time, Trichinella species were molecularly identified in gray wolves from Lithuania. Trichinella britovi was confirmed in all of the isolated Trichinella larvae using a multiplex PCR. Gray wolves in Lithuania may serve as a major source of zoonotic pathogens due to the presence of these parasites

First molecular characterization of Sarcocystis miescheriana in wild boars (Sus scrofa) from Latvia

Various muscle samples of wild boar (Sus scrofa) from Latvia were studied for the presence of Sarcocystis infection by means of morphological and molecular methods. Sarcocysts were detected in 122 out of 140 (87.1%) wild boar examined. According to the morphological appearance of sarcocysts, the observed cysts belonged to one morphological type and resembled Sarcocystis miescheriana. Twenty-three sarcocysts isolated from the muscles of Latvian wild boars were molecularly characterized at 18S rRNA, ITS1 and cox1. Additionally, eight sarcocysts obtained from Lithuanian wild boars were subjected to molecular analysis in order to compare intraspecific genetic variability. The amplified 18S rRNA region using newly designed primers is sufficiently variable to separate S. miecheriana from S. suihominis. All Latvian and Lithuanian isolates were confirmed belonging to S. miescheriana. No genetic variation was detected within 18S rRNA and ITS1. By contrast, the high intraspecific genetic variability of S. miescheriana was observed within cox1 since each newly obtained sequence represented a unique haplotype. The comparison made using S. miescheriana isolates from Italian and Japanese wild boar and Chinese domestic pig revealed the genetic similarity of the samples depending on their geographical distances. The current study provides the first detection of Sarcocystis infection in wild boars from Latvia and molecular characterization of S. miescheriana

<i>Sarcocystis</i> Species Richness in Sheep and Goats from Lithuania

Contradictory data is available on the intermediate host specificity of Sarcocystis spp. in farm animals. Therefore, the current work aimed at molecularly testing samples of sheep and goats reared in Lithuania to identify Sarcocystis species described in other intermediate hosts but suspected to be non-canonical parasites to these small ruminants. For this purpose, muscle samples from 47 domestic sheep and nine goats were examined. Sarcocystis species were identified using direct and nested PCR targeting cox1 and sequencing of positive amplified products. Along with the detection of the canonical Sarcocystis spp. in their respective intermediate hosts, the DNA of S. capracanis and S. morae was detected in sheep, although these species were previously thought to be specific to goats and deer, respectively. In addition, DNA from S. arieticanis and S. tenella was found in goats, even though these two species were believed to be sheep-specific. Notably, under light microscopy, only sarcocysts of S. capracanis specific to goats were observed. Thus, future research on the life cycle and host-specificity of Sarcocystis spp. examined is warranted

Novel luminescent dyes for confocal laser scanning microscopy used in Trematoda parasite diagnostics

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

Modeling the Distribution of the Chytrid Fungus <i>Batrachochytrium dendrobatidis</i> with Special Reference to Ukraine

Amphibians are the most threatened group of vertebrates. While habitat loss poses the greatest threat to amphibians, a spreading fungal disease caused by Batrachochytrium dendrobatidis Longcore, Pessier & D.K. Nichols 1999 (Bd) is seriously affecting an increasing number of species. Although Bd is widely prevalent, there are identifiable heterogeneities in the pathogen’s distribution that are linked to environmental parameters. Our objective was to identify conditions that affect the geographic distribution of this pathogen using species distribution models (SDMs) with a special focus on Eastern Europe. SDMs can help identify hotspots for future outbreaks of Bd but perhaps more importantly identify locations that may be environmental refuges (“coldspots”) from infection. In general, climate is considered a major factor driving amphibian disease dynamics, but temperature in particular has received increased attention. Here, 42 environmental raster layers containing data on climate, soil, and human impact were used. The mean annual temperature range (or ‘continentality’) was found to have the strongest constraint on the geographic distribution of this pathogen. The modeling allowed to distinguish presumable locations that may be environmental refuges from infection and set up a framework to guide future search (sampling) of chytridiomycosis in Eastern Europe

A Comparative Study of New Fluorescent Anthraquinone and Benzanthrone α-Aminophosphonates: Synthesis, Spectroscopy, Toxicology, X-ray Crystallography, and Microscopy of <i>Opisthorchis felineus</i>

In this research, we explore the synthesis of and characterize α-aminophosphonates derived from anthraquinone and benzanthrone, focusing on their fluorescence properties and potential applications in confocal laser scanning microscopy (CLSM). The synthesized compounds exhibit notable solvatochromic behavior, emitting fluorescence from green to red across various solvents. Spectroscopic analysis, including 1H-, 13C-, and 31P-NMR, FTIR, and mass spectrometry, confirms the chemical structures. The compounds’ toxicity is evaluated using etiolated wheat sprouts, revealing varying degrees of impact on growth and oxidative damage. Furthermore, the study introduces these α-aminophosphonates for CLSM imaging of the parasitic flatworm Opisthorchis felineus, demonstrating their potential in visualizing biological specimens. Additionally, an X-ray crystallographic study of an anthraquinone α-aminophosphonate provides valuable structural insights

A 38-year study on Trichinella spp. in wild boar (Sus scrofa) of Latvia shows a stable incidence with an increased parasite biomass in the last decade

Publisher Copyright: © 2015 Kirjušina et al.; licensee BioMed Central.Background: Trichinella spp. are zoonotic parasites transmitted to humans by the consumption of raw or insufficiently cooked meat of different animal species. The most common source of infection for humans is meat from pigs and wild boar (Sus scrofa). The aim of the present work was to evaluate the incidence of Trichinella spp. infections in wild boar hunted in Latvia over a 38 year interval (1976 to 2013). Methods: A total 120,609 wild boars were individually tested for Trichinella spp. by trichinoscopy and, in case of negativity, by artificial digestion of 25 g muscles, in the 1976-2005 period, and by artificial digestion of 25-50 g muscles in the 2006-2013 period. Trichinella spp. larvae were identified at the species level by multiplex PCR. Results: In the study period, the overall prevalence of infected wild boar was 2.5%. Trichinella britovi was the predominant (90%) species. The incidence of Trichinella spp. infection in wild boar exhibited two different trends. From 1976 to 1987, the incidence of infected/hunted wild boar increased from 0.23% to 2.56%, then it decreased to 0.19 in 1994. Thereafter, the incidence fluctuated between 0.05% and 0.37%. A statistically significant (P∈<∈0.05) correlation (r∈=∈0.54; p∈=∈0.0199) was found between the trend of Trichinella spp. incidence in hunted wild boar and the number of snow cover days from 1976 to 1993. From 1997 to 2013, the estimated wild boar population of Latvia increased by 4.9 times and the hunting bag by 9.7 times, with a stable incidence of Trichinella spp. in the population. It follows that the biomass of Trichinella spp. larvae and of T. britovi, in particular, increased. Conclusions: The incidence trends of Trichinella spp. in wild boar could be related to the role played by the snow in reducing the thermal shock and muscle putrefaction which increases the survival of the larvae in muscle tissues of carrion in the 1976-1993 period; and, in the 1997-2013 period, to the increased biomass of Trichinella spp. due to the increased carnivore populations, which are the main reservoirs of these parasites.publishersversionPeer reviewe