Ultrastructure of a haplosporidian containing Rickettsiae,associated with mortalities among cultured paua Haliotis iris

Uninucleate and multinucleate stages of a protozoan parasite are described from cultured abalone Haliotis iris Martyn, 1784 in New Zealand. The parasite is identified as a haplosporidian by the occurrence of multinucleate plasmodia, mitochondria with tubular cristae, lipid droplets, anastomosing endoplasmic reticulum (aER), multivesicular bodies (MVBs), haplosporogenesis by the production of haplosporosome-like bodies from nuclear membrane-bound Golgi, and their maturation to haplosporosomes. Coated pits occurred in the plasma membrane and coated vesicles were scattered in the cytoplasm, particularly in association with the Golgi face away from the nucleus, and aER. It is concluded that the outward face of the Golgi may be the trans face, and that aER is the trans-Golgi network. Coated pits and bristle-coated vesicles are reported from a haplosporidian for the first time. The vesicles in the MVBs resembled the cores and inner membranes of haplosporosomes, without the outer layer. The possible inter-relationships of these features are discussed. The abalone parasite differs from previously described haplosporidians in the apparent absence of a persistent mitotic spindle, and the presence of intracytoplasmic coccoid to rod-shaped bacteria resembling Rickettsiales-like prokaryotes. Phylogenetic analysis of the 16S rRNA gene sequence of the Rickettsiales-like prokaryotes indicated that these organisms belong to the Rickettsia cluster. The prokaryotes have a high (7%) sequence divergence from known Rickettsieae, with Rickettsia sp. and R. massiliae being the closest relatives. The lack of non-molecular evidence prevents us from proposing a new rickettsial genus at this time.     

Prokaryote infections in the New Zealand scallops Pecten novaezelandiae and Chlamys delicatula

Four intracellular prokaryotes are reported from the scallops Pecten novaezelandiae Reeve, 1853 and Chlamys delicatula Hutton, 1873. Elongated (1025 x 110 nm), irregular (390 x 200 nm), or toroidal (410 x 200 nm) mollicute-like organisms (M-LOs) occurred free in the cytoplasm in the digestive diverticular epithelial cells of both scallop species. Those in P. novaezelandiae bore osmiophilic blebs that sometimes connected the organisms together, and some had a rod-like protrusion, both of which resemble the blebs and tip structures of pathogenic mycoplasmas. The M-LOs in C. delicatula had a slightly denser core than periphery. Round M-LOs, 335 x 170 nm, occurred free in the cytoplasm of agranular haemocytes in P. novaezelandiae, without apparent harm to the host cell. In P. novaezelandiae, 2 types of highly prevalent (95 to 100%) basophilic inclusions in the branchial epithelium contained Rickettsia-like organisms (R-LOs). Type 1 inclusions occurred in moderately hypertrophied, intensely basophilic cells, 8 to 10 microm in diameter, containing elongate intracellular R-LOs, 2000 x 500 nm. Type 2 inclusions were elongated and moderately basophilic in markedly hypertrophic branchial epithelial cells, 50 x 20 microm in diameter, containing intracellular organisms 500 x 200 nm in diameter. The possible roles of these organisms in pathogenesis is discussed.     

Luminous vibriosis in rock lobster Jasus verreauxi (Decapoda: Palinuridae) phyllosoma larvae associated with infection by Vibrio harveyi

Studies were conducted to determine the cause of outbreaks of luminous vibriosis in phyllosoma larvae of the packhorse rock lobster Jasus verreauxi reared in an experimental culture facility. On 2 separate occasions mortalities of up to 75% over a period of 4 wk were observed in 4th to 5th and 8th to 10th instar phyllosomas at water temperatures of 20 and 23 degrees C, respectively. Affected larvae became opaque, exhibited small red spots throughout the body and pereiopods, and were faintly luminous when viewed in the dark. Histopathology showed that the gut and hepatopancreas tubules of moribund phyllosomas contained massive bacterial plaques. The hepatopancreas tubules of moribund larvae were atrophic and some contained necrotic cells sloughed into the lumen. Dense, pure cultures of a bacterium identified as Vibrio harveyi were isolated from moribund larvae. The disease syndrome was reproduced by in vivo challenge and V. harveyi was successfully reisolated from diseased larvae after apparently healthy larvae were exposed by immersion to baths of more than 10(4) V. harveyi ml(-1) at 24 degrees C. Injured larvae were more susceptible to infection than were healthy larvae. Survival of larvae experimentally and naturally exposed to V. harveyi was improved when antibiotics were administered via bath exposures.     

Ecology and welfare of aquatic animals in wild capture fisheries

The expansion of commercial aquaculture production has raised awareness of issues relating to the welfare of aquatic animals. The “Five Freedoms” approach to animal welfare was originally devised for farmed terrestrial animals, and has been applied in some countries to aquatic animals reared in aquaculture due to several commonalities inherent within food production systems. There are now moves towards assessing and addressing aquatic animal welfare issues that may arise in wild capture fisheries. However, all “five freedoms” are regularly contradicted in the natural environment, meaning this concept is inappropriate when considering the welfare of aquatic animals in their natural environments. The feelings-based approach to welfare relies on a suffering centered view that, when applied to the natural aquatic environment, requires use of value judgements, cannot encompass scientific uncertainty regarding awareness in fish, elasmobranchs and invertebrates (despite their unquestioned welfare needs), and cannot resolve the welfare conundrums posed by predator–prey interactions or anthropocentrically mediated environmental degradation. For these reasons, the feelings-based approach to welfare is inadequate, inappropriate and must be rejected if applied to aquatic animals in wild capture fisheries, because it demonstrably ignores empirical evidence and several realities apparent within the natural aquatic environment. Furthermore, application of the feelings-based approach is counterproductive as it can alienate key fisheries stakeholders, many of whom are working to address environmental issues of critical importance to the welfare, management and conservation of aquatic animal populations in their natural environment. In contrast, the function-based and nature-based approaches for defining animal welfare appear appropriate for application to the broad range of welfare issues (including emerging environmental issues such as endocrine disruption) that affect aquatic animals in their natural environment, without the need to selectively ignore groups such as elasmobranchs and invertebrates. We consider that the welfare needs of aquatic animals are inextricably entwined with the need for conservation of their populations, communities and their environment, an approach that is entirely consistent with the concept of ecosystem-based management.     

Dactylogyrids (Monogenoidea: Polyonchoinea) parasitising the gills of snappers (Perciformes: Lutjanidae): Species of Euryhaliotrema Kritsky & Boeger, 2002 from the golden snapper Lutjanus johnii (Bloch) off northern Australia, with a redescription of Euryhaliotrema johni (Tripathi, 1959) and descriptions of two new species

Three species of Euryhaliotrema Kritsky & Boeger, 2002 (Monogenoidea: Dactylogyridae) were collected from the gills of four golden snapper Lutjanus johnii (Bloch) (Lutjanidae) from the marine and brackish waters off Darwin, Northern Territory, Australia. Type-specimens of Ancyrocephalus johni Tripathi, 1959 apparently have not survived and the possibility existed that the species was based on specimens representing more than one species. Euryhaliotrema johni (Tripathi, 1959) (sensu Young, 1968) was redescribed and determined to most likely represent A. johni, originally described from the River Hooghly, Diamond Harbour, India. Two new species were described. Euryhaliotrema longibaculoides n. sp. was most similar to Euryhaliotrema longibaculum (Zhukov, 1976) Kritsky & Boeger, 2002 from Lutjanus spp. from the western Atlantic Ocean. It differed from E. longibaculum by having a male copulatory organ (MCO) with an elongate comparatively delicate shaft and a bulbous base (MCO U- or J-shaped with funnel-shaped base in E. longibaculum). Based on the comparative morphology of the haptoral sclerites, Euryhaliotrema lisae n. sp. was most similar to Euryhaliotrema cryptophallus Kritsky & Yang, 2012 from the gills of the mangrove red snapper Lutjanus argentimaculatus (Forsskål) from the South China Sea. Euryhaliotrema lisae differed from E. cryptophallus by having a copulatory complex with an obvious weakly sclerotised J-shaped MCO (MCO cryptic, delicate, and with a shaft comprising about one counterclockwise ring in E. cryptophallus).     

Perkinsus olseni in vitro isolates from the New Zealand clam Austrovenus stutchburyi

Perkinsus olseni infections are reported at 10%-84% prevalences among Austrovenus stutchburyi clams (cockles) in northern New Zealand coastal waters. However, P. olseni has not yet been propagated in vitro from New Zealand clams. In our sample of A. stutchburyi clams from Mangemangaroa Stream, New Zealand, 24% (8/34) showed low-intensity Perkinsus sp. infections among mantle and gill tissues incubated in alternative Ray's fluid thioglycollate medium (ARFTM), and 5% (4/79) showed Perkinsus sp. lesions by histological analyses. Among clams that were screened using a polymerase chain reaction (PCR) assay, 16% (3/19) were positive for Perkinsus sp. DNA. Alternative Ray's fluid thioglycollate medium-enlarged hypnospores from tissues of five infected clams yielded three in vitro Perkinsus sp. isolate cultures that were cloned before sequencing internal transcribed spacer (ITS) regions of their rRNA gene complex. For one isolate, ATCC PRA-205, large subunit (LSU) rRNA and actin genes were also sequenced. All nucleotide sequences from all isolates consistently identified them as P. olseni, as did their in vitro cell cycles and zoosporulation characteristics. All in vitro isolate cultures and their respective monoclonal derivative strains were cryopreserved and deposited for archiving and distribution by the American Type Culture Collection (http://www.atcc.org).     

Two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) from marine fishes off the northern coast of Australia

Based on light and electron microscopical studies, two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) are described from the ovary of marine perciform fishes off the northern coast of Australia (near Darwin): Philometra carangis n. sp. from the bluespotted trevally Caranx bucculentus Alleyne & Macleay (Carangidae) and P. carponotati n. sp. from the Spanish flag snapper Lutjanus carponotatus (Richardson) (Lutjanidae). Philometra carangis is mainly characterised by the length of the spicules (153–189 µm), the presence of a distinct dorsal protuberance consisting of two dorsolateral lamellar parts separated from each other by a smooth median field, a V-shaped mound on the male caudal extremity, a pair of large post-cloacal papillae and the body length of the males (3.22–4.15 mm). Philometra carponotati is distinguished from other congeneric species parasitising lutjanids by the length of the spicules and gubernaculum (225–252 and 99–117 µm, respectively), the absence of a dorsal protuberance on the distal lamellar part of the gubernaculum, the presence of a U-shaped mound on the male caudal extremity, a pair of large post-cloacal papillae and the body length of the male (3.74–4.31 mm). Besides the recently established Philometra zabidii Moravec & Diggles, 2014 (based on a single female), these two newly described nematodes are the only nominal gonad-infecting species of Philometra known to parasitise marine fishes in Australian waters.     

Development of a TaqMan PCR assay for the detection of Bonamia species

The development of molecular diagnostic assays with increased sensitivity compared with conventional histological techniques is highly desirable for effective management of bonamiosis in cultured oyster stocks and wild populations. A real-time TaqMan PCR assay was developed for the specific detection of Bonamia species in infected oyster tissues. The TaqMan assay was shown to be significantly more sensitive than histopathology. Although a real-time TaqMan PCR assay is comparable with conventional PCR in terms of sensitivity, it offers the advantages that it is a rapid test and has a very low risk of sample cross-contamination. Furthermore, it can be optimised to quantify the parasite load in samples. The assay detected Bonamia isolates from Australia, New Zealand, Europe, Canada, Chile and the USA and therefore demonstrated genus specificity as tested in this study.