In the last few years two factors have helped to significantly advance our understanding of the Myxozoa. First, the phenomenal increase in fin fish aquaculture in the 1990s has lead to the increased importance of these parasites; in turn this has lead to intensified research efforts, which have increased knowledge of the development, diagnosis. and pathogenesis of myxozoans. The hallmark discovery in the 1980s that the life cycle of Myxobolus cerebralis requires development of an actinosporean stage in the oligochaete. Tubifex tubifex, led to the elucidation of the life cycles of several other myxozoans. Also, the life cycle and taxonomy of the enigmatic PKX myxozoan has been resolved: it is the alternate stage of the unusual myxozoan, Tetracapsula bryosalmonae, from bryozoans. The 18S rDNA gene of many species has been sequenced, and here we add 22 new sequences to the data set. Phylogenetic analyses using all these sequences indicate that: 1) the Myxozoa are closely related to Cnidaria (also supported by morphological data); 2) marine taxa at the genus level branch separately from genera that usually infect freshwater fishes; 3) taxa cluster more by development and tissue location than by spore morphology; 4) the tetracapsulids branched off early in myxozoan evolution, perhaps reflected by their having bryozoan, rather than annelid hosts; 5) the morphology of actinosporeans offers little information for determining their myxosporean counterparts (assuming that they exist); and 6) the marine actinosporeans from Australia appear to form a clade within the platysporinid myxosporeans. Ribosomal DNA sequences have also enabled development of diagnostic tests for myxozoans. PCR and in situ hybridisation tests based on rDNA sequences have been developed for Myxobolus cerebralis, Ceratomyxa shasta, Kudoa spp., and Tetracapsula bryosalmonae (PKX). Lectin-based and antibody tests have also been developed for certain myxozoans, such as PKX and C. shasta. We also review important diseases caused by myxozoans, which are emerging or re-emerging. Epizootics of whirling disease in wild rainbow trout (Oncorhynchus mykiss) have recently been reported throughout the Rocky Mountain states of the USA. With a dramatic increase in aquaculture of fishes using marine netpens, several marine myxozoans have been recognized or elevated in status as pathological agents. Kudoa thyrsites infections have caused severe post-harvest myoliquefaction in pen-reared Atlantic salmon (Salmo salar), and Ceratomyxa spp., Sphaerospora spp., and Myxidium leei cause disease in pen-reared sea bass (Dicentrarchus labrax) and sea bream species (family Sparidae) in Mediterranean countries.
A rod-shaped, gill-associated virus (GAV) was found to be associated with mortalities of cultured adult Penaeus monodon from 4 farms in Queensland, Australia, from January to June 1996. Infected prawns were observed swimming at the surface and edges of ponds and displayed varying degrees of red body colouration. The lymphoid organs of infected prawns appeared disorganized, were devoid of normal tubule structure and contained highly eosinophilic areas which corresponded to foci of highly infected and necrotic cells. Rod-shaped, enveloped virions and helical nucleocapsids were identified in lymphoid organ and gill cells. Nucleocapsids were 166-435 nm X 16-18 nm and enveloped virions were 183-200 nm X 34-42 nm. GAV isolated from infected prawns collected from 3 farms was successfully transmitted to healthy adult P, rnonodon, resulting in mortality from 7 to 8 d postinoculation. Mortality also occurred in prawns infected experimentally with GAV derived from lymphoid organs and gills. The morphology and cytopathology of GAV closely resemble that observed for lymphoid organ virus (LOV) from Australia and yellow-head virus (YHV) from Thailand. Molecular data are required to determine the phylogenetic relationships and appropriate taxonomic classification of these 3 prawn viruses.
Subadult Penaeus monodon shrimp were sampled from 4 farms in Queensland, Australia. Histological observations on lymphoid organs showed the formation of abnormal cell foci, which resembled tubules lacking a central hemolymph vessel. These formations contained hypertrophied nuclei with marginated chromatin, vacuolated cells and inclusion bodies which stained positive with Feulgen's reaction. Male and female F! monodon broodstock were sampled from 1 farm and showed the same abnormal histological features within the lymphoid organ as the subadults. Electron microscopy revealed tightly enveloped, cylindrical particles, 163-200 nm X 36-63 nm, packed into paracrystalline arrays within the cytoplasm and infrequently within the nucleus. Arrays of virions were also observed within the cytoplasm of broodstock gill cells. Cross-sections of the particles revealed electron dense nucleic acid cores and envelopes. Nucleocapsids, 83-590 nm X 13-15 nm, were seen free and in vesicles within the cytoplasm. A negatively stained preparation of partially purified lymphoid organ revealed a nucleocapsid with one conical end. Lymphoid organ virus (LOV) resembles yellow-head virus (YHV) from Thailand in its morphology and cytopathology and rhabdovirus of penaeid shrimp (RPS) from the Americas in its cytopathology.
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