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Parasitology Research Oct 2012The spore morphology and molecular systematic of a new microsporidian which was isolated from the common sea bream Pagrus pagrus (F: Sparidae Linnaeus, 1758) from the...
The spore morphology and molecular systematic of a new microsporidian which was isolated from the common sea bream Pagrus pagrus (F: Sparidae Linnaeus, 1758) from the Red Sea, Egypt have been studied. Fifty-six out of 300 (18.7%) of this fish were infected with microsporidian parasites. The infection was appeared as whitish, ellipsoid, round, or elongated nodules embedded in the epithelial lining of the peritoneum and also in the intestinal epithelium. Light microscopic study revealed that nodules were encapsulated by a fibrous layer encircling numerous mature spores measuring 1.7 ± 0.6 (1.5-2.7 μm) × 1.5 ± 0.3 μm (1.2-1.8 μm) in size. Ultrastructure of spores was characteristic for the genus Pleistophora: dimorphic, uninucleate spores (each spore possesses three to five polar filament coils) and a posterior vacuole. Also, the early recognizable stages of the parasite within nodules include uninucleated, binucleated, and multinucleated meronts followed by detachment of the plasmalemma of the sporont producing sporoblasts which mature to spores that consist of a spore coat and spore contents. Also, we analyzed the small subunit ribosomal gene (SSUrDNA) using PCR and sequencing specimens from the marine populations of P. pagrus fish from the Red Sea. From blast searches, sequence analysis, and phylogenetic analysis, we did not find corresponding GenBank entries to our species. Comparison of the nucleotide sequences showed that the sequence of our microsporidium was most similar to five Pleistophora species with degrees of identity (>91.5%). It was most similar (97.8% identity) to that of Pleistophora hyphessobryconis (account no. GU126672) differing in 19 nucleotide positions and with lower divergence value, Pleistophora ovariae (96.2% identity, account no. AJ252955), Pleistophora hippoglossoideos (91.9% identity, account no. AJ252953), Pleistophora mulleri (91.9% identity, account no. EF119339), and Pleistophora typicalis (91.9% identity, account no. AJ252956). So, they likely represent new species named Pleistophora pagri sp. n. with accession number JF797622 and a GC content of 53%.
Topics: Animals; Base Composition; Cluster Analysis; DNA, Fungal; DNA, Ribosomal; Egypt; Fish Diseases; Genes, rRNA; Histocytochemistry; Microscopy; Microsporidiosis; Molecular Sequence Data; Perciformes; Phylogeny; Pleistophora; Prevalence; RNA, Fungal; RNA, Ribosomal, 18S; Sequence Analysis, DNA; Spores, Fungal
PubMed: 22773009
DOI: 10.1007/s00436-012-3012-9 -
Journal of Fish Biology Jun 2012A transparent mutant tiger barb Puntius tetrazona was identified and characterized by its transparent body, which allows clear visualization of internal organs....
A transparent mutant tiger barb Puntius tetrazona was identified and characterized by its transparent body, which allows clear visualization of internal organs. Hybridization of this mutant with the albino variant produces a transparent and albinoid double phenotype, and the transparency of this mutant is controlled by a recessive allele. Light microscopic and ultrastructural examinations show that in contrast to normal individuals, transparent mutants lack iridophores, and light penetrates unimpeded through the body. Pleistophora sp. infection was observed in vivo, allowing live observation of parasite dissemination and the consequent pathological alterations in the fish body as well as the simultaneous acquisition of data on the dynamics and spatial pattern of pathogenic invasion. It is superior to common fish models, as dynamic experimental data can be obtained from individual fish.
Topics: Albinism; Animals; Chromatophores; Cyprinidae; Fish Diseases; Microsporidia; Microsporidiosis; Mutation; Pigmentation
PubMed: 22650423
DOI: 10.1111/j.1095-8649.2012.03280.x -
Parasitology Research Jul 2012Pleistophora dammami sp. n. is described from Saurida undosquamis from the Arabian Gulf in Saudi Arabia. Infection appeared as whitish cysts in the intestinal wall....
Pleistophora dammami sp. n. is described from Saurida undosquamis from the Arabian Gulf in Saudi Arabia. Infection appeared as whitish cysts in the intestinal wall. Cysts ranged in size from 1 to 4 mm. The prevalence of the infection across both fish sexes was 17.5% (24/420). Two kinds of spores were recognized, microspores and macrospores, and each were ovoid in shape. The microspores measured ~2.5 × 2.0 μm in size, while the macrospores measured ~6.0 × 3.0 μm. Ultrastructurally, the parasite did not form xenoma but it formed cysts surrounded by thick cyst wall. All stages of development as meronts, sporonts, sporoblast and spores occurred in the cytoplasm of the host cells within sporophorous vesicles. The stages of development occurred asynchronously and thus all stages were randomly distributed within the cysts. Meronts were elliptical and multinucleated, with unpaired nuclei which constantly divided giving rise to new sporonts. During the transition to sporonts, the border of the meronts increased in thickness to form dense discontinuous cell coat. Later, the sporont divided into sporoblast cells which gradually differentiated the typical organelles of the spores. In mature spores, the polar filament was arranged in 20-24 coils in two rows either side of the posterior vacuole. All ultrastructural and morphological criteria indicate that the described species belongs to the genus Pleistophora.
Topics: Animals; Chordata; Female; Fish Diseases; Gastrointestinal Diseases; Intestines; Male; Marine Biology; Pleistophora; Prevalence; Saudi Arabia; Spores, Fungal
PubMed: 22374307
DOI: 10.1007/s00436-012-2855-4 -
Emerging Infectious Diseases Sep 2011The Phylum Microsporidia comprises >1,200 species, only 15 of which are known to infect humans, including the genera Trachipleistophora, Pleistophora, and Brachiola. We...
The Phylum Microsporidia comprises >1,200 species, only 15 of which are known to infect humans, including the genera Trachipleistophora, Pleistophora, and Brachiola. We report an infection by Tubulinosema sp. in an immunosuppressed patient.
Topics: Acute Kidney Injury; Aged; Fatal Outcome; Female; Humans; Immunocompromised Host; Leukemia, Lymphocytic, Chronic, B-Cell; Microsporidia; Muscle, Skeletal; Myositis
PubMed: 21888805
DOI: 10.3201/eid1709.101926 -
Parasitology Research Feb 2012One hundred three out of 225 (45.8%) of the Red Sea fish Saurida tumbil were infected with microsporidian parasites. The infection was recorded as tumor-like masses...
One hundred three out of 225 (45.8%) of the Red Sea fish Saurida tumbil were infected with microsporidian parasites. The infection was recorded as tumor-like masses (whitish macroscopic cysts) or xenomas often up to 2 cm in diameter and embedded in the peritoneal cavity. Generally, the infection was increased during winter 63.8% (86 out of 135) and fall to 18.9% (17 out of 90) in summer. Light microscopic study revealed that xenomas were encapsulated by a fibrous layer encircling numerous sporophorous vesicles filled with mature spores measuring 1.7 ± 0.6 (1.5-2.7 μm) × 1.5 ± 0.3 μm (1.2-1.8 μm) in size. Ultrastructural microscopic study showed the presence of smooth membranes of the sarcoplasmic reticulum forming a thick, amorphous coat surrounding various developmental stages of the parasite. The various recognizable stages of the parasite were uninuclear, binucleated, and multinucleated meronts followed by detachment of the plasmalemma of the sporont from the sporophorous vesicle producing sporoblasts. Mature spores consist of a spore coat and spore contents. The spore contents consist of the uninucleated sporoplasm and a posterior vacuole located at the posterior end. The polar tube consists of a straight shaft and a coiled region (26-32 coils) arranged in many rows along the inside periphery of the spore. The polaroplast consisted of an anterior region of closely and loosely packed membranes. Molecular analysis based on the small subunit rDNA gene was performed to determine the phylogenetic position of the present species. The percentage identity between this species and a range of other microsporidia predominantly from aquatic hosts demonstrated a high degree of similarity (>92%) with eight Pleistophora species. Comparison of the nucleotide sequences and divergence showed that the sequence of the present microsporidium was most similar to that of Pleistophora anguillarum (99.8% identity) differing in 13 nucleotide positions. So, the present species was recorded and phylogenetically positioned as a new species of Pleistophora.
Topics: Animals; Chordata; Cluster Analysis; DNA, Fungal; DNA, Ribosomal; Fish Diseases; Genes, rRNA; Histocytochemistry; Incidence; Indian Ocean; Microscopy; Microsporidiosis; Molecular Sequence Data; Phylogeny; Pleistophora; RNA, Fungal; RNA, Ribosomal, 18S; Seasons; Sequence Analysis, DNA; Spores, Fungal
PubMed: 21858478
DOI: 10.1007/s00436-011-2597-8 -
Diseases of Aquatic Organisms Jun 2011Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or...
Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation, or small spores in nuclei (i.e. Nucleospora salmonis). Hence, a variety of selective stains that enhance visualization of spores is recommended. We discovered that the Luna stain, used to highlight eosinophils, red blood cells, and chitin in arthropods and other invertebrates, also stains spores of Pseudoloma neurophilia. We compared this stain to the Gram, Fite's acid fast, Giemsa, and H&E stains on 8 aquatic microsporidian organisms that were readily available in our 2 laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum, and an unidentified microsporidian from UK mitten crabs Eriocheir sinensis. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the 2 microsporidia from invertebrates: S. mytilovum and the unidentified microsporidian from E. sinensis.
Topics: Animals; Brachyura; Coloring Agents; Fishes; Microsporidia; Mycoses; Mytilus; Spores, Fungal; Staining and Labeling
PubMed: 21848126
DOI: 10.3354/dao02346 -
Journal of Medical Microbiology Apr 2011Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron...
Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron microscopy. This study describes a single SYBR Green real-time PCR assay for the simultaneous detection and species identification of such infections. This assay was highly sensitive, routinely detecting infections containing 400 parasites (g stool sample)(-1), whilst species identification was achieved by differential melt curves on a Corbett Life Science Rotor-Gene 3000. A modification of the QIAamp DNA tissue extraction protocol allowed the semi-automated extraction of DNA from stools for the routine diagnosis of microsporidial infection by real-time PCR. Of 168 stool samples routinely analysed for microsporidian spores, only five were positive by microscopy. By comparison, 17 were positive for microsporidial DNA by real-time analysis, comprising 14 Enterocytozoon bieneusi, one Encephalitozoon cuniculi and two separate Pleistophora species infections.
Topics: Benzothiazoles; DNA, Fungal; Diamines; Encephalitozoon cuniculi; Enterocytozoon; Humans; Microsporidiosis; Molecular Sequence Data; Organic Chemicals; Pleistophora; Polymerase Chain Reaction; Quinolines; Sensitivity and Specificity; Sequence Analysis, DNA; Staining and Labeling; Transition Temperature
PubMed: 21183599
DOI: 10.1099/jmm.0.026781-0 -
Parasitology Research Jan 2011Out of 600 marine fish from the Red Sea belonging to three different species that were collected and examined for microsporidian parasites, 87 (14.5%) fish were found to...
Out of 600 marine fish from the Red Sea belonging to three different species that were collected and examined for microsporidian parasites, 87 (14.5%) fish were found to be infected. The infection was recorded as cysts or xenomas embedded in the gut epithelium and the peritoneal cavity of the three fish species. The highest percent of infection with microsporidian parasites was recorded in Saurida tumbil 19.5% (39/200) followed by Pagrus pagrus 15% (45/300) and the lowest percent of infection was recorded in Epinephelus chlorostigma 3% (three out of 100). After rupture of the cysts, the spores were released and examined by light microscopy. Each spore was elongated to ellipsoidal in shape and possessed a posterior vacuole which is characteristic to phylum Microspora. They measure 1.6 ± 0.5 μm (1.5-2.4 μm) × 1.3 ± 0.1 μm (1.3-2.0 μm) in Saurida tumbil and Pagrus pagrus, respectively. The spores of Pleistophora sp recorded from E. chlorostigma were ovoid to pyriform in shape and measure 1.9 ± 0.5 μm (1.8-2.7 μm) × 1.6 ± 0.4 μm (1.5-2.4 μm).
Topics: Animals; Fish Diseases; Fishes; Histocytochemistry; Indian Ocean; Intestinal Mucosa; Microscopy; Microsporidia; Microsporidiosis; Peritoneal Cavity; Prevalence
PubMed: 20865429
DOI: 10.1007/s00436-010-2061-1 -
Diseases of Aquatic Organisms Jul 2010Zebrafish Danio rerio are important models for biomedical research, and thus, there is an increased concern about diseases afflicting them. Here we describe infections...
Zebrafish Danio rerio are important models for biomedical research, and thus, there is an increased concern about diseases afflicting them. Here we describe infections by Pleistophora hyphessobryconis (Microsporidia) in zebrafish from 3 laboratories. As reported in other aquarium fishes, affected zebrafish exhibited massive infections in the skeletal muscle, with no involvement of smooth or cardiac muscle. In addition, numerous spores within macrophages were observed in the visceral organs, including the ovaries. Transmission studies and ribosomal RNA (rRNA) gene sequence comparisons confirmed that the parasite from zebrafish was P. hyphessobryconis as described from neon tetra Paracheirodon innesi. Ten 15 d old zebrafish were exposed to P. hyphessobryconis collected from 1 infected neon tetra, and 7 of 10 fish became infected. Comparison of P. hyphessobryconis small subunit rRNA gene sequence from neon tetra with that obtained from zebrafish was nearly identical, with < 1% difference. Given the severity of infections, P. hyphessobryconis should be added to the list of pathogens that should be avoided in zebrafish research facilities, and it would be prudent to avoid mixing zebrafish used in research with other aquarium fishes.
Topics: Animals; DNA, Ribosomal; Fish Diseases; Microsporidia; Microsporidiosis; Phylogeny; Zebrafish
PubMed: 20853741
DOI: 10.3354/dao02245 -
Infection, Genetics and Evolution :... Oct 2010Microsporidia comprise an unusual group of intracellular, eukaryotic parasites that exhibit ubiquitous distribution throughout the animal kingdom. We analysed the small...
Molecular characterisation of the Microsporidia of the amphipod Gammarus duebeni across its natural range revealed hidden diversity, wide-ranging prevalence and potential for co-evolution.
Microsporidia comprise an unusual group of intracellular, eukaryotic parasites that exhibit ubiquitous distribution throughout the animal kingdom. We analysed the small subunit ribosomal gene (SSUrDNA) using PCR and sequencing and screened 894 Gammarus duebeni (Crustacea, Amphipoda) specimens from 35 European marine and freshwater populations. We discovered considerable hidden microsporidian diversity. Blast searches, sequence analysis and phylogenetic analysis revealed intraspecific sequence variants of known species Dictyocoela duebenum,Dictyocoela muelleri, Pleistophora mulleri and Nosema granulosis. For seven SSUrDNA sequences, we did not find corresponding GenBank entries; they likely represent new species, provisionally classified within the genus Microsporidium. Phylogenetic reconstructions revealed their position as polyphyletic, thereby lending support to the hypothesis of an early microsporidian radiation within this host group. Nevertheless, four of the presumptive novel species formed a discrete and well-supported subclade in the phylogenetic analysis. The respective host specimens were collected from disjunct freshwater sites in Wales, Ireland and Brittany (France) and may represent a new, G. duebeni-specific, microsporidian genus. At the population level, our screening showed that parasitism through Microsporidia is the rule rather than the exception in G. duebeni. We found Microsporidia in 91% of sampled G. duebeni populations. This finding may have considerable consequences for the interpretation of results from ecological, behavioural, physiological and evolutionary studies of the host, as parasitism can significantly influence these traits. Because the host G. duebeni has a complex phylogeography and evolutionary history, the studied host-parasite system may have potential as a model system for investigating processes of co-evolution.
Topics: Amphipoda; Animals; DNA, Fungal; Demography; Evolution, Molecular; Genetic Variation; Host-Parasite Interactions; Microsporidia; Phylogeny
PubMed: 20601176
DOI: 10.1016/j.meegid.2010.06.011