The lung-dwelling nematode Rhabdias engelbrechti sp. n. was found in five of eight examined banded rubber frogs in Limpopo Province, South Africa. The species is differentiated from species of Rhabdias Stiles et Hassall, 1905 occurring in the Afrotropical Realm based on the presence of a globular cuticular inflation at the anterior end, the buccal capsule walls being distinctly divided into anterior and posterior parts, the buccal capsule size (6-9 μm × 16-18 μm), and the body length (3.8-6.1 mm). Rhabdias engelbrechti is the tenth species of the genus found in Afrotropical anurans. Our molecular phylogenetic analysis based on the complete sequences of the ITS region and partial sequences of large subunit (28S) gene of the nuclear ribosomal RNA demonstrates that the new species is more closely related to the Eurasian species Rhabdias bufonis (Schrank, 1788) than to two other species from sub-Saharan Africa represented in the tree. In addition, partial sequences of the mitochondrial protein coding cox1 and ribosomal 12S genes of the new species have shown significant differences from all previously published sequences of these genes from African species of Rhabdias., Yuriy Kuzmin, Ali Halajian, Sareh Tavakol, Wilmien J. Luus-Powell, Vasyl V. Tkach., and Obsahuje bibliografii
Monoclonal antibody specific for an epitope of cretory-secretory antigen protein of Opisthorchis felineus (Rivolta, 1884) (Trematoda: Opisthorchiidae) with a molecular weight of 28 kDa was used in a sandwich enzyme-linked immunosorbent assay (ELISA) for immobilisation of liver fluke specific antigen to the solid phase. Examination of human sera by this ELISA compared with commercial assays demonstrated that the monoclonal antibody epitope is located within this significant parasite protein. Anti-idiotypic antibody specific for the paratope of this monoclonal antibody was obtained by a hybridoma technique. Mimicking an epitope of excretory-secretory antigen of O. felineus, it had the capacity to bind specific antibody and elicit an antibody response. The value of anti-idiotypic antibody as a substitute for the liver fluke antigen was tested by ELISA using serum samples of infected dogs. Anti-idiotypic antibody proved to be of value in both an indirect-ELISA and a competitive-ELISA for diagnosis of opisthorchiasis. Mature trematodes were isolated from all infected animals. The faecal egg counts were negative in dogs with a relatively small number of parasites, despite finding antibodies in serum by ELISA. Substitution of parasite antigen with anti-idiotype avoids the use of experimental animals and also reduces time-consuming steps of antigen preparation., Aitbay K. Bulashev, Sergey N. Borovikov, Shynar S. Serikova, Zhanbolat A. Suranshiev, Vladimir S. Kiyan, Saule Z. Eskendirova., and Obsahuje bibliografii
Tissue samples from wildlife from South Africa were opportunistically collected and screened for haemoprotozoan parasites using nonspecific PCR primers. Samples of 127 individuals were tested, comprising over 50 different species. Haemogregarines were the most commonly identified parasites, but sarcocystids and piroplasmids were also detected. Phylogenetic analyses estimated from the 18S rDNA marker highlighted the occurrence of several novel parasite forms and the detection of parasites in novel hosts. Phylogenetic relationships, which have been recently reviewed, appear to be much more complex than previously considered. Our study highlights the high diversity of parasites circulating in wildlife in this biodiverse region, and the need for further studies to resolve taxonomic issues., D. James Harris, Ali Halajian, Joana L. Santos, Lourens H. Swanepoel, Peter John Taylor, Raquel Xavier., and Obsahuje bibliografii
Faecal samples from the rock hyrax (Procavia capensis jayakari Thomas) were collected from the Ibex Reserve in central Saudi Arabia. Eimerian oocysts, which are believed to represent a new species described here as Eimeria tamimi sp. n., were detected in 40 out of 93 samples. Oocysts were fully sporulated in 24-48 hours at 25 ± 2 °C. Sporulated oocysts of E. tamimi sp. n. were ovoid, measuring 35-42 × 19-25 μm (39 × 23 μm), a length/width ratio 1.5-2 (1.7). Oocyst wall was bilayered and measured 1.5 μm in thickness. Micropyle, oocyst residuum and polar granules were not present. Sporocysts are elongate, measuring 12-18 × 9-12 μm (15 × 10 μm), with a length/width ratio 1.1-1.8 (1.5) prominent Stieda bodies and sporocyst residuum. Experimental infection of two clinically healthy rock hyraxes with sporulated oocysts of E. tamimi sp. n. resulted in shedding unsporulated oocysts 5-10 days post infection. Partial sequences of 18S ribosomal RNA (18S rDNA) and cytochrome C oxidase subunit 1 (COI) regions were amplified using the polymerase chain reaction (PCR) and sequenced. Phylogenetic analysis based on 18S rDNA using maximum likelihood (ML) and Bayesian inference (BI) methods revealed that E. tamimi sp. n. grouped with Eimeria quokka Barker, O'Callaghan et Beveridge, 1988, E. mundayi Barker, O´Callaghan et Beveridge, 1988, E. potoroi Barker, O'Callaghan et Beveridge, 1988 and E. gaimardi Barker, O'Callaghan et Beveridge, 1988 marsupials. Eimerian species have been regarded as a paraphyletic group and the present investigation confirmed the conflict between phenotypic traits, used widely in the classification of this group of parasites., Osama B. Mohammed, Manei M. Aljedaie, M. S. Alyousif and Nabil Amor., and Obsahuje bibliografii
The record is confirmed of a “Clariobranchus” hybrid Clarias gariepinus × Heterobranchus sp. specimen in the River Danube in Croatia. Clarias gariepinus was introduced to Europe for farming and research purposes. Because of their faster growth and short time to achieve market size, “Clariobranchus” hybrids have gained attention in European aquaculture. To date, no record has been found in the literature of “Clariobranchus” hybrid escapees in inland waters of Europe. The present finding suggests that the “Clariobranchus” hybrid escaped from an unprotected aquaculture system or open pond farm. The threat of establishment of a population of this hybrid in suitable environments, such as thermal ponds, cannot be ruled out.
In the parthenogenetic monogeneans of the genus Gyrodactylus Nordmann, 1832, the genetic diversity within or between hosts is determined by the relative roles of lateral transmission and clonal propagation. Clonality and limited transmission lead to high-amplitude metapopulation dynamics and strong genetic drift. In Baltic populations of the three-spined stickleback Gasterosteus aculeatus Linnaeus, the local mitochondrial diversity of Gyrodactylus arcuatus Bychowsky, 1933 is very high, and spatial differentiation weak. To understand the transmission dynamics in a single location, the transmission of the parasite from adults to next generation sticklebacks was investigated in a northern Baltic brackish water location. By sequencing 777 nt of cox1, as many as 38 separate mitochondrial haplotypes were identified. In August, the intensity of gyrodactylid infection on adult hosts was high, the haplotype diversity (h) was extreme and differentiation between fish was negligible (total h = 0.926, mean h = 0.938). In October, only 46% of the juvenile sticklebacks carried G. arcuatus. The number of parasites per young fish followed a Poisson distribution 0.92 ± 1.04 (mean ± SD) on October 2, and was clearly overdispersed 2.38 ± 5.00 on October 25. The total haplotype diversity of parasites on juveniles was nearly as high as in adults (h = 0.916), but the mean per fish was only h = 0.364 (FST = 0.60), due to low intensity of infection and rapid clonal propagation of early arrivals. The initial first come first served advantage of the first gyrodactylid colonisers will be lost during the host adulthood via continuous transmission. Nesting and polygamy are suggested as factors maintaining the high genetic diversity of the parasite population. The transmission dynamics and, consequently, the population structure of Baltic G. arcuatus is fundamentally different from that of G. salaris Malmberg, 1957, on the Baltic salmon Salmo salar Linnaeus., Jaakko Lumme, Marek S. Ziętara., and Obsahuje bibliografii
Myxobolus filamentum sp. n. was found infecting gill filaments of three of 39 Brycon orthotaenia Günther specimens examined (8%), which were taken from the river São Francisco in Minas Gerais state, Brazil. Plasmodia of the parasite were white and long, measuring 5 mm in lenght. Mature spores of M. filamentum sp. n. were oval from the frontal view and biconvex from the lateral view, measuring 7.5-9.7 µm (9.0 ± 0.3 µm) in length and 5.2-7.3 µm (6.2 ± 0.4 µm) in width. The polar capsules were elongated and equal in size, measuring 3.8-5.5 µm (4.7 ± 0.3 µm) in length and 1.3-2.2 µm (1.7 ± 0.1 µm) in width. The development of the parasite led to compression of the adjacent tissues and inflammatory infiltrate with granulocytic cells. Ultrastructural observation revealed that the plasmodia were delimited by two membranes, which had numerous and extensive pinocytotic channels extending into the wide ectoplasm zone. The plasmodial wall exhibited abundant villi-like projections and a thin layer of granular material prevented direct contact between the plasmodial wall and the host tissue. Phylogenetic analysis, based on 18S rDNA, showed M. filamentum sp. n. as a sister species of Myxobolus oliveirai Milanin, Eiras, Arana, Maia, Alves, Silva, Carriero, Ceccarelli et Adriano, 2010, a parasite of other fish species of the genus Brycon Müller et Troschel from South America., Juliana Naldoni, Suellen A. Zatti, Kassia R.H. Capodifoglio, Tiago Milanin, Antônio A.M. Maia, Marcia R.M. Silva, Edson A. Adriano., and Obsahuje bibliografii
Hexokinase (HXK) is the first key enzyme in the glycolytic pathway and plays an extremely important role in energy metabolism. By searching the microsporidian database, we found a sequence (NBO_27g0008) of Nosema bombycis Nägali, 1857 with high similarity to hexokinase-2, and named it as NbHXK2. The NbHXK2 gene has 894 bp and encodes 297 amino acids with 34.241 kD molecular weight and 5.26 isoelectric point. NbHXK2 contains 31 phosphorylation sites and 4 potential N-glycosylation sites with signal peptides and no transmembrane domain. Multiple sequence alignment showed that NbHXK2 shares more than 40% amino acid identity with that of other microsporidia, and the homology with hexokinase-2 of Nosema tyriae Canning, Curry, Cheney, Lafranchi-Tristem, Kawakami, Hatakeyama, Iwano et Ishihara, 1999, Nosema pyrausta (Paillot, 1927) and Nosema ceranae Fries, Feng, da Silva, Slemenda et Pieniazek, 1996 was 89.17%, 87.82% and 69.86%, respectively. Phylogenetic analysis based on the amino acid sequence of hexokinase showed that all microsporidia cluster together in the same clade, and are far away from animals, plants and fungi, and that N. bombycis is closely related to N. tyriae; N. pyrausta; N. ceranae and Nosema apis Zander, 1909. Immunolocalisation with the prepared polyclonal antibody showed that NbHXK2 was mainly distributed in the cytoplasm and plasmalemma in proliferative, sporulation stage and mature spore of N. bombycis. qRT-PCR assay showed that the NbHXK2 expressed at higher level during spore germination and at early stage of proliferation. These results indicate that N. bombycis may use its own glycolytic pathways to supply energy for infection and development, especially germination and in the early stage of proliferation, and acquire energy from the host through certain ways as well.
The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.
Parasitic organisms inhabiting the alimentary canal should permanently resist the destructive action of host digestive enzymes. The intestinal parasites were shown to produce specific protease inhibitors protecting them from proteolysis. However, little is known about this adaptive mechanism in cestodes so far, especially for the tapeworms dwelling inside the fish intestines. Here, we explored the ability to inactivate proteolytic enzymes in the fish tapeworm Eubothrium rugosum (Batsch, 1786) (Bothriocephalidea) parasitising the intestine of wild burbot, Lota lota (Linnaeus). The assays were conducted with different concentrations of commercial trypsin and homogenate of intestinal mucosa both being the sources of proteinases. The incubation of live E. rugosum in trypsin solutions of two different concentrations caused a significant decrease in the enzyme activity. The extent of activity reduction was dependent on trypsin concentration. At the same time, the inhibitory effect of the worm incubation medium turned out to be statistically insignificant. These findings suggest partial adsorption of the enzyme to the tegument surface, with its further inactivation. In contrast to the incubation medium, the worm extract suppressed over 80% of trypsin activity and nearly half of the proteolytic activity in the mucosa homogenate. Notably, the inhibitory activity of the tapeworms hardly depended on their size characteristics. Finally, the research has demonstrated secretion of proteinase inhibitor in E. rugosum, which appears to be essential for its survival in enzymatically hostile environment., Galina I. Izvekova, Tatyana V. Frolova, Evgeny I. Izvekov., and Obsahuje bibliografii