Denitsa Teofanova

Geographic distribution modelling for ruminant liver flukes (Fasciola hepatica) in south-eastern Europe.

Maximum entropy ecological niche modelling was utilised to predict the geographic range for fluke genotypes and haplotypes in south-eastern Europe, using the Maxent program. The lowest (0.832) and the highest (0.947) area under the curve values were observed in the models for the haplotypes CtCmt1 and CtCmt2.2, respectively. Precipitation and temperature contribute equally to model building of the genotypes based on the 28S rDNA gene. In regard to the mtDNA gene region, precipitation is the most important factor in modelling the CtCmt1 haplotype range, while temperature appears to be the most important factor in modelling the CtCmt2.1 and CtCmt2.2 haplotype ranges. The highest level of probability for the geographic distribution of Fasciola hepatica genotypes and haplotypes covered the regions of southern Bulgaria and central and northern Greece which contain a high concentration of potential ruminant hosts.

Genetic diversity of liver flukes (Fasciola hepatica) from Eastern Europe

The genetic diversity of liver fluke populations in three different countries from Eastern Europe (Greece, Bulgaria, and Poland) was determined and compared with available data from other countries. Specifically, SNPs from regions of two nuclear genes, 28S rDNA, β-tubulin 3 and an informative region of the mitochondrial genome were examined. Two major lineages for the 28S rDNA gene based on the highly polymorphic 105th nucleotide position were found. These lineages were widely and almost equally spread not only through the countries studied but also in other investigated geographical areas. Two basic lineages and additional haplotypes were defined for the mtDNA gene region which consisted of the cytochrome c oxidase subunit III gene, transfer RNA histidine gene and cytochome b gene. The basic lineages were observed within Greek, Bulgarian, and Polish Fasciola hepatica populations but the distribution of additional haplotypes differed between the populations from the three countries. For the β-tubulin 3 gene multiple polymorphic sites were revealed but no explicit clades. The SNPs were spread unequally in all studied geographical regions with an evident distinction between the Greek and Polish specimens. Additional genotypes for the 28S rDNA region as well as haplotypes of the mtDNA region that were typical for the Greek or Polish populations were observed. Significant polymorphisms for β-tubulin 3 gene were displayed with decreasing percentage of presence within populations from Greece to Poland. There was an amino acid substitution in β-tubulin 3 protein found only among Polish specimens. It is hypothesized that genotypic differences between Greek, Bulgarian, and Polish liver fluke populations are due to territorial division and genetic drift in past epochs.

Haplotype diversity in autochthonous Balkan cattle breeds

The Bulgarian Grey cattle (BGC) and theShorthorn Rhodopean cattle (SRC) are Balkan indigenousbreeds that are part of the Podolian and Brachicerous(Busha) cattle groups respectively. Concerning the mtDNAdiversity of Balkan cattle breeds, previous studies focusedon Busha cattle and Grey cattle populations.

A Novel Secretory Poly-Cysteine and Histidine-Tailed Metalloprotein (Ts-PCHTP) from Trichinella spiralis (Nematoda)

Background Trichinella spiralis is an unusual parasitic intracellular nematode causing dedifferentiation of the host myofiber. Trichinella proteomic analyses have identified proteins that act at the interface between the parasite and the host and are probably important for the infection and pathogenesis. Many parasitic proteins, including a number of metalloproteins are unique for the nematodes and trichinellids and therefore present good targets for future therapeutic developments. Furthermore, detailed information on such proteins and their function in the nematode organism would provide better understanding of the parasite - host interactions. Methodology/Principal Findings In this study we report the identification, biochemical characterization and localization of a novel poly-cysteine and histidine-tailed metalloprotein (Ts-PCHTP). The native Ts-PCHTP was purified from T. spiralis muscle larvae that were isolated from infected rats as a model system. The sequence analysis showed no homology with other proteins. Two unique poly-cysteine domains were found in the amino acid sequence of Ts-PCHTP. This protein is also the first reported natural histidine tailed protein. It was suggested that Ts-PCHTP has metal binding properties. Total Reflection X-ray Fluorescence (TXRF) assay revealed that it binds significant concentrations of iron, nickel and zinc at protein:metal ratio of about 1∶2. Immunohistochemical analysis showed that the Ts-PCHTP is localized in the cuticle and in all tissues of the larvae, but that it is not excreted outside the parasite. Conclusions/Significance Our data suggest that Ts-PCHTP is the first described member of a novel nematode poly-cysteine protein family and its function could be metal storage and/or transport. Since this protein family is unique for parasites from Superfamily Trichinelloidea its potential applications in diagnostics and treatment could be exploited in future.

SIGNIFICANCE OF MILK PROTEIN GENES POLYMORPHISM FOR BULGARIAN RHODOPEAN CATTLE: COMPARATIVE STUDIES

ABSTRACT The aim of the present research was to reveal the genotype profile of the local Bulgarian Rhodopean cattle population with respect to αs1-casein, kappa-casein andβ-lactoglobulin genes, by PCR-RFLP assay and conformational 2D PAGE. According to these profiles, it is possible to determine the association between each genotype and milk qualitative and quantitative traits and to establish the position of the breed with regard to the genetic diversity other European cattle breeds. The investigation also revealed the high significance of the genetic variants of the three above-mentioned milk protein genes for: Clarification of the influence of other breeds on the Bulgarian Rhodopean cattle population; Obtaining data for genetic drift among the Bulgarian Rhodopean cattle population, the indigenous Shorthorn Rhodopean cattle population as a predecessor breed and the Jersey cow as a main improvement breed; Gathering the necessary information about preservation of this local breed, which is important for the Bulgarian biodiversity gene fund.

Application of Milk Proteins Genetic Polymorphism for Selection and Breeding of Dairy Cows in Bulgaria

© 2012 Hristov et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Application of Milk Proteins Genetic Polymorphism for Selection and Breeding of Dairy Cows in Bulgaria

Issues Associated with Genetic Diversity Studies of the Liver Fluke, Fasciola Heptica (Platyhelminthes, Digenea, Fasciolidae)

Parasitic diseases are huge problem for human and veterinary medicine and for economy, agriculture and wildlife management. One of these diseases is fasciolosis, which is caused by two trematode species, Fasciola hepatica (liver fluke) and Fasciola gigantica. Only F. hepatica is a concern in Europe and Americas but the distribution of both species overlaps in many areas of Africa and Asia (Mas-Coma et al., 2005). F. hepatica occurs mostly in cattle, sheep and wild ruminants. Recently, worldwide losses in animal productivity due to fasciolosis were estimated at over US

Genotyping of Endemic for Rhodopa Mountains Shorthorn Rhodopean Cow Breed

3.2 billion per annum (Spithill et al., 1999). The infection of humans has been regarded as accidental for many years. However, fasciolosis is now recognized as an emerging human disease: the World Health Organization (WHO, 2006) has estimated that 2.4 million people are infected with F. hepatica and a further 180 million are at risk of infection (Mas-Coma et al., 1999).

Native and Recombinant Fatty Acid Binding Protein 3 from Fasciola Hepatica as a Potential Antigen

ABSTRACT Shorthorn Rhodopean cattle breed is one of two endemic for Bulgaria cow breeds. The breed is characterized by high vitality and fertility, calving ease, resistance to diseases, and low requirements for feed. Milk yield is low, but fat and protein content of milk are high. Therefore it is of high significance to reveal the genotype profile of that cattle population which represents the aim of present study. 38 animals of that breed were genotyped for two lactoprotein genes: kappa-casein and alfaS1-casein through PCR-RFLP analysis. The results for kappa- casein locus show slight superiority of heterozygous AB genotype and prevalence of uncommon B allele. Analysis of polymorphism of alfaS1-casein gene displays that about 53% of the animals are with homozygous BB genotype, about 34% - with heterozygous BC genotype and only ∼13% (5 cows)—with homozygous CC genotype. It may be concluded that Shorthorn rhodopean cow breed is with specific genotype profile similar to other cattle population in South-Eastern Europe.

Mitochondrial DNA haplotype analysis of liver fluke in bison from Bialowieza Primaeval Forest indicates domestic cattle as the likely source of infection

ABSTRACT Lipid-binding proteins are members of widely distributed protein family. They take part in metabolism of different lipophilic ligands and also have a role in physiological activity, metabolism and disposition of essential hydrophobic compounds (fatty acids, phospholipids, eicosanoids, retinoids, etc.) which are important molecules involved in several cellular processes including gene transcription, immune responses etc. Fatty acid binding proteins (FABPs) are mostly cytosolic but several members are excretory-secretory (E-S) proteins. There is a lot of data that helminthic FABPs from E/S products cause antigenic and allergenic reactions. The aim of the present study is to make a detailed investigation of native and produced recombinant FABP3 as a potential antigen for vaccine creation. The native FABP3 from Fasciola hepatica was purified and recombinant one was expressed in E. coli. Polyclonal antibody was produced against recombinant FABP3 and it was tested on the native one. Obtained results give the opportunity to investigate the effectiveness of the native and recombinant FABP3 as an immunotherapeutic agent and if it could be used for immunoprophilactic.