Kudoa prunusi n. sp. (Myxozoa: Multivalvulida) from the brain of Pacific bluefin tuna Thunnus orientalis (Temminck & Schlegel, 1844) cultured in Japan

Kudoa prunusi n. sp. (Myxozoa; Multivalvulida) is described from the brain of Pacific bluefin tuna Thunnus orientalis cultured in Japan. Numerous white cysts, up to 0.5mm in size, were found on and in the brain. Spores having typically five spore valves and five polar capsules resembled a five-petal cherry blossom in apical view and were conical shape with a round bottom in side view. Average spore size was 9.63 (8.5-10.3) μm in width and 7.50 (6.7-8.6) μm in length. The spore dimensions of K. prunusi overlapped with those of Kudoa yasunagai ex Sillago ciliata having five to six spore valves, but they were clearly distinct in spore shape, 18S rDNA and 28S rDNA sequences (0.3% and 1.7% differences, respectively). Phylogenetic analysis of 18S rDNA revealed that K. prunusi grouped with the brain-infecting multivalvulid species, K. yasunagai, K. chaetodoni, K. lethrini and K. neurophila, rather than five-valved Kudoa spp. Combined with morphological, molecular and biological differences, K. prunusi was proven to be a new species.     

Two unusual myxozoans,Kudoa quadricornis n. sp. (Multivalvulida) from the muscle of goldspotted trevally (Carangoides fulvoguttatus) and Kudoa permulticapsula n. sp. (Multivalvulida) from the muscle of Spanish mackerel (Scomberomorus commerson) from the Great Barrier Reef,Australia

Two unusual myxozoan parasites are described from the somatic muscle of 2 reef fishes from Australia's Great Barrier Reef. Kudoa quadricornis n. sp. from the somatic muscle of Carangoides fulvoguttatus is morphologically consistent with other Kudoa sp., having 4 polar capsules and 4 shell valves. Kudoa quadricornis n. sp. is unique in that it has a pyriform spore body with a greater length than width (7.82-9.95 and 5.94-8.66 microm, respectively) and distinct posterolateral projections. Spores of Kudoa permulticapsula n. sp. observed within pseudocysts of the somatic muscle tissue of Scomberomorus commerson are different from those of all other myxozoans. The ovoid spores (length, 4.69-6.65 microm; width, 8.42-9.92 microm; thickness, 6.36-8.33 microm) contain 13 polar capsules with an equal number of shell valves. Phylogenetic analysis using small subunit ribosomal DNA sequences of K. quadricornis n. sp. and K. permulticapsula n. sp. showed that these parasites cluster within a clade comprised of Kudoa species. This brings into question the division of parasites of the Multivalvulida into genera based solely on polar capsule numbers.     

First report of three Kudoa species from eastern Australia: Kudoa thyrsites from mahi mahi (Coryphaena hippurus), Kudoa amamiensis and Kudoa minithyrsites n. sp. from sweeper (Pempheris ypsilychnus)

Fish species around the world are parasitized by myxozoans of the genus Kudoa, several of which infect and cause damage of commercial importance. In particular, Kudoa thyrsites and Kudoa amamiensis infect certain cultured fish species causing damage to muscle tissue, making the fish unmarketable. Kudoa thyrsites has a broad host and geographic range infecting over 35 different fish species worldwide, while K. amamiensis has only been reported from a few species in Japanese waters. Through morphological and molecular analyses we have confirmed the presence of both of these parasites in eastern Australian waters. In addition, a novel Kudoa species was identified, having stellate spores, with one polar capsule larger than the other three. The SSU rDNA sequence of this parasite was 1.5% different from K. thyrsites and is an outlier from K. thyrsites representatives in a phylogenetic analysis. Furthermore, the spores of this parasite are distinctly smaller than those of K. thyrsites, and thus it is described as Kudoa minithyrsites n. sp. Although the potential effects of K. minithyrsites n. sp. on its fish hosts are unknown, both K. thyrsites and K. amamiensis are associated with flesh quality problems in some cultured species and may be potential threats to an expanding aquaculture industry in Australia.     

Brain infecting kudoids of Australia's coral reefs, including a description of Kudoa lemniscati n. sp. (Myxosporea: Kudoidae) from Lutjanus lemniscatus (Perciformes: Lutjanidae) off Ningaloo Reef, Western Australia

A survey of the myxosporean fauna of Australian marine fishes revealed the presence of a number of putative species of Kudoidae (Multivalvulida) forming pseudocysts between the outer meningeal layer and the outer surface of the brains of the lutjanids Caesio cuning, Lutjanus carponotatus, Lutjanus ehrenbergii and Lutjanus fulviflamma and the mugilid Liza vaigiensis from Lizard Island on the Great Barrier Reef, Australia and Lutjanus lemniscatus off Ningaloo Reef, Western Australia. Morphometric data combined with Bayesian inference and maximum likelihood analyses of small subunit (SSU) and large subunit (LSU) ribosomal DNA (rDNA) was used for species identification and to explore relationships among these taxa. The brain-infecting taxa examined here formed a well-supported clade to the exclusion of non-brain infecting species in the phylogenetic analyses. The combined diagnostic approach identified an undescribed taxon, Kudoa lemniscati n. sp., from the brain of L. lemniscatus (Perciformes: Lutjanidae) off Ningaloo Reef, Western Australia, which we describe and characterise here. K. lemniscati n. sp. can be distinguished from all other species of Kudoa based on the combination of the distinct tropism for forming pseudocysts in the brain tissue, spores with 7 or 8 equal shell valves and 7 or 8 polar capsules, spore size and the differences in the SSU and LSU rDNA sequence data relative to other kudoids. Kudoa chaetodoni was found in the lutjanids C. cuning and L. carponotatus, expanding the known host range for this species to include chaetodontids and lutjanids. L. ehrenbergii and L. fulviflamma were infected with Kudoa lethrini off Lizard Island, a parasite previously known only from lethrinids. Specimens putatively identified as Kudoa yasunagai from Liza vaigiensis and Lutjanus ehrenbergii were morphologically similar and genetically identical over the SSU rDNA dataset to previously reported specimens, but differed by 4 to 11 nucleotides over the LSU dataset from the remaining isolates examined here. While these data are not definitive, they suggest the presence of a K. yasunagai complex.     

Kudoa hexapunctata n. sp. (Myxozoa: Multivalvulida) from the somatic muscle of Pacific bluefin tuna Thunnus orientalis and re-description of K. neothunni in yellowfin tuna T. albacares

Since Kudoa septempunctata in olive flounder (Paralichthys olivaceus) was indicated to cause food poisoning in humans, other Kudoa species are suspected to have pathogenic potential. Recently, a myxosporean possibly associated with food poisoning in humans consuming raw Pacific bluefin tuna, Thunnus orientalis, was identified as Kudoa neothunni. This is a known causative myxosporean of post-harvest myoliquefaction in yellowfin tuna Thunnus albacares. Regardless of the significant differences in the 28S rDNA sequence and the pathological character (with/without myoliquefaction) between the two T. orientalis and T. albacares isolates, they were considered intraspecific variants of K. neothunni. However, the light and low-vacuum electron microscopic observations in the present study revealed that there were two morphotypes; pointed- and round-type spores, which were significantly differentiated by the ratio of suture width to spore width. Furthermore, the two morphotypes were genetically distinguishable by the 28S rDNA sequence analysis. This morphological and molecular evidence validates that the two Kudoa types are separate species, and thus the pointed- and round-types are referred to as K. neothunni and Kudoa hexapunctata n. sp., respectively. K. neothunni was detected solely from T. albacares, whereas K. hexapunctata n. sp. was found not only from T. orientalis but also from T. albacares.     

Two multivalvulid myxozoans causing postmortem myoliquefaction: Kudoa megacapsula n. sp. from red barracuda (Sphyraena pinguis) and Kudoa thyrsites from splendid alfonso (Beryx splendens)

Postmortem myoliquefaction associated with multivalvulid myxozoans was found in fillets of red barracuda (Sphyraena pinguis) and splendid alfonso (Beryx splendens), which were imported to Japan from China and South Africa, respectively. Morphological examinations of the myxozoans from the somatic muscle of red barracuda revealed that spores (30.3-44.7 microm in maximum thickness) had 4 distinct winglike valves, in which 1 extremely large (12.7 x 5.8 microm), 2 small, and 1 vestigial polar capsule were present. The small subunit ribosomal DNA sequence analysis showed that the myxozoan cluster within a clade was composed of Kudoa thyrsites, Kudoa minithyrsites, and Kudoa lateolabracis, all having stellate spores with 1 polar capsule larger than the other 3. On the basis of these characteristics, we describe this parasite as Kudoa megacapsula n. sp. Morphological and molecular analyses of the myxozoan from splendid alfonso identified it as K. thyrsites, which has been described from many marine fishes. To our knowledge, this is the first record of K. thyrsites in splendid alfonso.     

Kudoa hypoepicardialis n. sp. (Myxozoa: Kudoidae) and associated lesions from the heart of seven perciform fishes in the northern Gulf of Mexico

Kudoa hypoepicardialis n. sp. infects the space between the epicardium and the compact myocardium and, in intense infections, the pericardial chamber of man-of-war fish (Nomeus gronovii) (Nomeidae) (the type host), blue runner (Caranx crysos) (Carangidae), Warsaw grouper (Epinephelus nigritus) (Serranidae), Atlantic tripletail (Lobotes surinamensis) (Lobotidae), northern red snapper (Lutjanus campechanus) (Lutjanidae), black drum (Pogonias cromis) (Sciaenidae), and bluefish (Pomatomus saltatrix) (Pomatomidae) in the northern Gulf of Mexico. This is the first report of a Kudoa sp. from the heart of a fish in the Gulf of Mexico, and of these hosts, only the bluefish was previously identified as a host for a species of Kudoa. Spores of the new species varied slightly in size among these hosts but were regarded as conspecific based on their nearly identical (99.9%) small-subunit (SSU) ribosomal DNA (rDNA) sequence. The new species differs both from the 4 nominal species of Kudoa reported from fishes in the Gulf of Mexico and from K. pericardialis, an allopatric species that infects the pericardial cavity, by the combination of having a large spore, a small polar capsule, and a polar filament with a single coil. The new species is morphologically and genetically most similar to K. shiomitsui, an allopatric species that infects the heart and pericardial cavity, but is distinguished from it based on a 4.2% difference in the SSU rDNA sequence. Heart lesions primarily were restricted to the vicinity of plasmodia and included a layer of fibrinous inflammation characterized by lymphocytes, macrophages, and granulomas as well as epithelioid encapsulations around plasmodia. Heavily infected hosts had melanin-like deposits and adipose cells beneath the epicardium. and the epicardium was discontinuous and apparently breached by plasmodia in some regions. Cardiac muscle, gill, liver, spleen, intestine, and kidney were normal.     

Multivalvulid myxozoans from eastern Australia: three new species of Kudoa from scombrid and labrid fishes of the Great Barrier Reef, Queensland, Australia

Three new species of Kudoa, each having 6 polar capsules, are described from the somatic muscle of fishes collected on the Great Barrier Reef, Queensland, Australia. Kudoa grammatorcyni n. sp. was observed in the shark mackerel Grammatorcynus bicarinatus. Spores are stellate in apical view, width (all measurements in microm) 8.62 (8.03-8.95); thickness 8.14 (7.63-8.68); suture width 7.7 (7.24-8.16); length 6.54 (6.32-6.71); polar capsule length 3.68 (3.55-3.82); polar capsule width 1.72 (1.65-1.84). Kudoa scomberomori n. sp. is described from the Spanish mackerel Scomberomorus commerson. Spores are stellate in apical view, width 7.56 (6.84-8.16); thickness 6.79 (6.18-7.63); suture width 5.92 (5.26-6.32); length 5.43 (5.00-6.18); polar capsule length 3.24 (3.03-3.55); polar capsule width 1.37 (1.25-1.51). Kudoa thalassomi n. sp. is described from the moon wrasse Thalassoma lunare. Spores are stellate in apical view, width 10.66 (9.47-11.84); thickness 9.37 (8.55-10.79); suture width 7.98 (6.84-8.82); length 6.65 (6.18-7.11); polar capsule length 4.92 (4.74-5.00); polar capsule width 2.12 (2.04-2.24). All 3 species differ in spore morphology from the 1 previously described myxozoan with 6 polar capsules, Hexacapsula neothunni from yellowfin tuna Neothunnus macropterus, which has since been reassigned to Kudoa.     

Life history, pathology, and description of Kudoa ovivora n. sp. (Myxozoa, Myxosporea): an ovarian parasite of Caribbean labroid fishes

We describe Kudoa ovivora n. sp. from ovaries of bluehead wrasse, Thalassoma bifasciatum, and record its presence in 6 species (Labroidei) collected in the San Blas Islands. Panama. Kudoa ovivora spores are quadrate with rounded edges in apical view, oval-shaped with apical valve extensions in side view (mean spore dimensions: length 6.5 microns, width 7.7 microns, thickness 6.9 microns; mean polar capsule dimensions: length 2.1 microns, width 1.5 microns). This is the first Kudoa species from gonads of fishes. Prevalence of infection varied among labrids (Thalassoma bifasciatum, Halichoeres bivittatus, Halichoeres garnoti, Halichoeres poevi), with T. bifasciatum exhibiting the greatest prevalence. Density of infection, measured as percent infected eggs, also varied among species with highest densities occurring in H. garnoti. Kudoa ovivora may not require an intermediate host because fishes fed infected tissue developed more infections than unfed fish. Infected eggs are inviable and larger and heavier than uninfected eggs. Infected eggs contain more organic and inorganic material, indicating that K. ovivora increases resource allocation to eggs. Therefore, infected females may have reduced growth, fecundity, and/or spawning activity. Because males were uninfected and all identified hosts are protogynous sequential hermaphrodites, further studies of K. ovivora may provide new insights on the costs/benefits of sex change.     

Systemic infection of Kudoa lutjanus n. sp. (Myxozoa: Myxosporea) in red snapper Lutjanus erythropterus from Taiwan

A new species of Kudoa lutjanus n. sp. (Myxosporea) is described from the brain and internal organs of cultured red snapper Lutjanus erythropterus from Taiwan. The fish, 260 to 390 g in weight, exhibited anorexia and poor appetite and swam in the surface water during outbreaks. Cumulative mortality was about 1% during a period of 3 wk. The red snapper exhibited numerous creamy-white pseudocysts, 0.003 to 0.65 cm (n = 100) in diameter, in the eye, swim bladder, muscle and other internal organs, but especially in the brain. The number of pseudocysts per infected fish was not correlated with fish size or condition. Mature spores were quadrate in apical view and suboval in side view, measuring 8.2 +/- 0.59 microm in width and 7.3 +/- 0.53 microm in length. The 4 valves were equal in size, each with 1 polar capsule. Polar capsules were pyriform in shape, measuring 3.62 +/- 0.49 microm in length and 2.2 +/- 0.49 microm in width. Mild inflammatory responses or liquefaction of host tissue were associated with K. lutjanus n. sp. infection. The junction of shell valves appeared as overlapping, straight lines. The polar filament formed 2 to 3 coils. A general PCR (polymerase chain reaction) primer for Kudoa amplified the small subunit (SSU) rDNA sequences, and the amplified gene was sequenced. It was evident from the phylogenetic tree that the 3 strains tested, AOD93020M, AOD93028M and AOD93028B, were identical and belonged to the Kudoa SS rRNA subgroup. The evolutionary tree showed that these strains form a unique clade, at a distance from other Kudoa species and myxosporeans. The spore's morphological and ultrastructural characteristics, as well as the SS rDNA properties of the isolates, were also essentially identical and served to distinguish them from representative Kudoa. It is, therefore, proposed that the strains isolated from the diseased red snapper be assigned to a new species.     

Kudoa iwatai (Myxosporea: Multivalvulida) in wild and cultured fish in the Red Sea: redescription and molecular phylogeny

Gilt-head sea bream, Sparus aurata L., the Mediterranean's most important mariculture species, has been cultured for the last 30 yr in Eilat (Israeli Red Sea). Kudoa sp. was the first myxosporean parasite reported from this species. In recent years, an increase in prevalence in both land-based and sea-cage facilities in Eilat has been observed. Infections with the same Kudoa species appeared in cultured European sea bass Dicentrarchus labrax (L.) and grey mullet Mugil cephalus in the same farms, as well as in 10 species of wild Red Sea reef fish, indicating that Kudoa sp. is not fastidious with regard to its host. All affected species displayed 1- to 2-mm (up to 5 mm) whitish, spherical, or oval polysporous plasmodia. The parasite established multiple site infections, most commonly in the muscles and intracranial adipose tissue of the brain and eye periphery. Other sites were subcutaneous adipose tissue, nerve axons, mouth, eye, mesenteries, peritoneum, swim bladder, intestinal musculature, heart, pericardium, kidney, and ovary. On the basis of spore morphology, the parasite was identified as Kudoa iwatai Egusa and Shiomitsu, 1983. Ultrastructural features were comparable to those of previously studied Kudoa species. The 18S rDNA from 7 Red Sea isolates was sequenced and compared with the sequence of the same gene from K. iwatai isolated from cultured red sea bream, Pagrus major, in Japan. The phylogenetic position of K. iwatai within the genus was determined using sequence analysis of all related taxa available in GenBank. The 3 isolates of K. iwatai clustered together on a newly formed, highly supported clade. The Red Sea strain of K. iwatai is apparently native to the region. In the absence of records of this Kudoa sp. from the extensive Mediterranean sea bream and sea bass production industries, introduction with its Mediterranean hosts seems unlikely. Therefore, we conclude that K. iwatai is an Indo-Pacific species that, in the Red Sea, has extended its host range to include the allochthonous gilt-head sea bream, European sea bass, and grey mullet.     

Myxobolus albi n. sp. (Myxozoa) from the Gills of the Common Goby Pomatoschistus microps Krøyer (Teleostei: Gobiidae)

ABSTRACT. A recent investigation into the myxozoan fauna of common gobies, Pomatoschistus microps , from the Forth Estuary in Scotland, revealed numerous myxosporean cysts within the gill cartilage. They were composed of polysporous plasmodia containing myxobolid spores that were morphologically different from the other known species of Myxobolus and from the myxosporeans previously recorded from this host (i.e. the ceratomyxid Ellipsomyxa gobii , infecting the gall bladder, and the kudoid Kudoa camarguensis , infecting the muscle tissues). Spores were ovoid, 9.4 × 9.1 μm with a thickness of 6.6 μm, with two pyriform polar capsules, the polar filaments of which had four to five turns. Molecular analysis of the parasite's small subunit rDNA region, based upon a contiguous sequence of 1,558 base pairs, discriminated it from other myxosporean species that have been characterized so far. A comparison of the spore morphology and the molecular sequences determined for this new isolate with other myxozoans described to date, confirmed its identity as a previously unknown myxobolid supporting the proposal that this isolate be elevated to the species level as a new species within the genus Myxobolus . A phylogenetic analysis places this new myxobolid, Myxobolus albi n. sp., in a basal position of a clade containing the majority of Henneguya spp. sequenced to date and various Myxobolus spp.     

Host specificity and local infection dynamics of Kudoa leptacanthae n. sp. (Multivalvulida: Kudoidae) from the pericardial cavity of two Zoramia spp. (Perciformes: Apogonidae) at Lizard Island lagoon, Queensland, Australia

Kudoa leptacanthae n. sp. was identified within the pericardial cavities of two apogonid species, Zoramia leptacantha and Zoramia viridiventer, from waters off Lizard Island on the Great Barrier Reef. The species shows a close taxonomic affinity, both morphologically and genetically, to Kudoa shiomitsui Egusa & Shiomitsu 1983 reported from a tetraodontid and scombrid, both from Japan. The infection was at high prevalences (average=75.8%±0.63, n=343) within schools of the two host species. Different sporogonic stages ('developing' and 'fully-developed') were observed in pseudocysts within the pericardium of the hosts. Kudoa leptacanthae n. sp. appears to be a persistent infection within the Zoramia species hosts. No seasonal variation was detected for this host/parasite system and the parasite did not affect host condition, when assessed using host length/weight ratio as a coarse indicator. We hypothesise that a source of infection in close proximity to the home patch reef of apogonid schools may lead to the high prevalence and intensity of infection reported in this study.     

Intra-specific variation of Kudoa spp. (Myxosporea: Multivalvulida) from apogonid fishes (Perciformes), including the description of two new species, K. cheilodipteri n. sp. and K. cookii n. sp., from Australian waters

Kudoa spp. from the musculature and intestinal mucosa of species of the teleost family Apogonidae were examined for their taxonomic identity. Two novel species are characterised: Kudoa cheilodipteri n. sp. from the musculature of Cheilodipterus quinquelineatus Cuvier, Ostorhinchus cyanosoma (Bleeker) and O. aureus (Lacépède); and Kudoa cookii n. sp. from the submucosa of the intestines of O. cookii (Macleay) only. Both species are characterised using morphology, small subunit ribosomal DNA (SSU rDNA), large subunit ribosomal DNA (LSU rDNA), and biological characters. Three new host records, O. cyanosoma, O. aureus and Apogon doederleini, and associated geographical, morphological and genetic data are also provided for Kudoa whippsi Burger & Adlard, 2010. Morphological and molecular intra-specific variation of all isolates assigned to K. whippsi is also examined. Phylogenetic analyses further support the idea that tissue tropism is a distinguishing character between morphologically similar species; species reported here display close relatedness to morphologically similar species infecting the same tissue within their hosts.     

A new species of myxozoan (Myxosporea) from the brain and spinal cord of rainbow trout (Oncorhynchus mykiss) from Idaho

A new species of Myxosporea, Myxobolus neurotropus n. sp., is described from the brain and spinal cord of rainbow trout (Oncorhynchus mykiss) from Duncan Creek, Owyhee County, Idaho. Spores are oval, have 2 pyriform polar capsules, and possess a thick spore wall (sutural rim) with a short intracapsular offshoot. The mean spore dimensions are length 11.8 microm, width 10.8 microm, and thickness 8.8 microm. This myxozoan is compared to other described Myxobolus species found in cranial tissues of salmonids in terms of spore morphology and phylogenetic analysis. Because it is found in brain and spinal cord, it is encountered while performing screening tests for Myxobolus cerebralis, the causative agent of salmonid whirling disease. Where chronic inflammation and granulomatous lesions are associated with M. cerebralis, histological examination shows no host response to M. neurotropus n. sp. A diagnostic polymerase chain reaction (PCR) test is included as an aid in properly identifying the species.     

Diagnostic polymerase chain reaction assay to detect Kudoa neurophila (Myxozoa: Multivalvulida) in a marine finfish hatchery

A single-round polymerase chain reaction (PCR) diagnostic assay was developed from a small subunit ribosomal DNA (SSU rDNA) gene sequence to detect the myxozoan parasite Kudoa neurophila, the causative agent of myxozoan disease in the hatchery reared marine finfish, striped trumpeter Latris lineata (Forster). The assay was developed for use as a disease control management tool in a hatchery system specifically designed to research and produce marine finfish such as striped trumpeter juveniles for aquaculture. The assay is sufficiently species specific and sensitive enough to detect a small fragment of the parasite's SSU rDNA. At the lower limits of detection, the test is consistently positive to an estimated 0.1 spore or 60 fg of parasite DNA per 25 microl PCR reaction in serial dilution and positive to an estimated 0.1 spore in 25 mg of infected fish CNS tissue (4 spores g(-1). Specifically, the test is capable of detecting early stages of the life cycle within the fish host and consequently diagnosing an infection not normally detected using traditional histological techniques. The test is also effective for screening water supplies and prey species cultures throughout the hatchery system to determine bio-security efficacy, to assist in identification of an alternate or other primary fish host, to indicate the location of potential disease reservoirs, and to enable a targetted approach to disease prevention.     

Phylogenetic relationships amongst Chloromyxum Mingazzini, 1890 (Myxozoa: Myxosporea), and the description of six novel species from Australian elasmobranchs

Six novel species of Chloromyxum Mingazzini, 1890 are described using a whole evidence approach combining morphometric and molecular data, together with features of their biology. Elasmobranchs were collected in Australian waters, from the Great Barrier Reef, Queensland, off Lizard and Heron Islands; from Moreton Bay, southeast Queensland; off Hobart, Tasmania; and from the Tamar River, Launceston, Tasmania. The novel species proposed here are: Chloromyxum hemiscyllii n.sp. from Hemiscyllium ocellatum; Chloromyxum kuhlii n.sp. from Neotrygon kuhlii; Chloromyxum lesteri n.sp. from Cephaloscyllium laticeps; Chloromyxum mingazzinii n.sp. from Pristiophorus nudipinnis; Chloromyxum myliobati n.sp. from Myliobatis australis; and Chloromyxum squali n.sp. from Squalus acanthias. A seventh species from Squalus acanthias is also reported but due to limited material is not formally described. Molecular phylogenetic analyses revealed that the genus Chloromyxum is polyphyletic, and species from elasmobranchs form a well-supported sister clade, with the type species Chloromyxum leydigi, to all other congeneric species clustering within the freshwater myxosporean clade. Morphological analysis showed that elasmobranch-infecting species are predominantly pyriform shaped, have clearly thickened spore apex and possess caudal filaments, compared to other Chloromyxum species which are generally spherical or subspherical, and lack caudal filaments. These morphological and phylogenetic data provide further support for the erection of new genera, but we conservatively consider the species described in this study and other elasmobranch-infecting Chloromyxum species as Chloromyxum sensu strictu, whilst the freshwater teleost infecting and amphibian infecting species we will assign as Chloromyxum sensu lato, until more comprehensive data are available.     

Description of Babesia duncani n.sp. (Apicomplexa: Babesiidae) from humans and its differentiation from other piroplasms

The morphologic, ultrastructural and genotypic characteristics of Babesia duncani n.sp. are described based on the characterization of two isolates (WA1, CA5) obtained from infected human patients in Washington and California. The intraerythrocytic stages of the parasite are morphologically indistinguishable from Babesia microti, which is the most commonly identified cause of human babesiosis in the USA. Intraerythrocytic trophozoites of B. duncani n.sp. are round to oval, with some piriform, ring and ameboid forms. Division occurs by intraerythrocytic schizogony, which results in the formation of merozoites in tetrads (syn. Maltese cross or quadruplet forms). The ultrastructural features of trophozoites and merozoites are similar to those described for B. microti and Theileria spp. However, intralymphocytic schizont stages characteristic of Theileria spp. have not been observed in infected humans. In phylogenetic analyses based on sequence data for the complete18S ribosomal RNA gene, B. duncani n.sp. lies in a distinct clade that includes isolates from humans, dogs and wildlife in the western United States but separate from Babesia sensu stricto, Theileria spp. and B. microti. ITS2 sequence analysis of the B. duncani n.sp. isolates (WA1, CA5) show that they are phylogenetically indistinguishable from each other and from two other human B. duncani-type parasites (CA6, WA2 clone1) but distinct from other Babesia and Theileria species sequenced. This analysis provides robust molecular support that the B. duncani n.sp. isolates are monophyletic and the same species. The morphologic characteristics together with the phylogenetic analysis of two genetic loci support the assertion that B. duncani n.sp. is a distinct species from other known Babesia spp. for which morphologic and sequence information are available.     

A new myxozoan parasite from the Amazonian fish Metynnis argenteus (Teleostei, Characidae): light and electron microscope observations

Myxobolus metynnis n. sp. (Phylum Myxozoa) is described in the connective subcutaneous tissues of the orbicular region of the fish, Metynnis argenteus (Characidae), collected in the lower Amazon River, near the city of Peixe Boi, Pará State, Brazil. Polysporic, histozoic plasmodia were delimited by a double membrane with numerous microvilli on the peripheral cytoplasm. Several life-cycle stages, including mature spores, were observed. An envelope formed by numerous fine and anastomosed microfibrils was observed at the spore surface. The spore body presented an ellipsoidal shape and was about 13.1 microm long, 7.8 microm wide, and 3.9 microm thick. Elongated-pyriform polar capsules were of equal size, measuring 5.2 microm in length, 3.2 microm in width, and possessing a polar filament with 8-9 turns around the longitudinal axis. The binucleated sporoplasm contained a vacuole and numerous sporoplasmosomes. These were circular in cross-section, showing an adherent eccentric, dense structure, with a half-crescent section. Based on the morphological differences and host specificity, we propose that the parasite is a new species named Myxobolus metynnis n. sp.