Hatching Strategies in Monogenean (Platyhelminth) Parasites that Facilitate Host Infection

In parasites, environmental cues may influence hatching of eggs and enhance the success of infections. The two major endoparasitic groups of parasitic platyhelminths, cestodes (tapeworms) and digeneans (flukes), typically have high fecundity, infect more than one host species, and transmit trophically. Monogeneans are parasitic flatworms that are among the most host specific of all parasites. Most are ectoparasites with relatively low fecundity and direct life cycles tied to water. They infect a single host species, usually a fish, although some are endoparasites of amphibians and aquatic chelonian reptiles. Monogenean eggs have strong shells and mostly release ciliated larvae, which, against all odds, must find, identify, and infect a suitable specific host. Some monogeneans increase their chances of finding a host by greatly extending the hatching period (possible bet-hedging). Others respond to cues for hatching such as shadows, chemicals, mechanical disturbance, and osmotic changes, most of which may be generated by the host. Hatching may be rhythmical, larvae emerging at times when the host is more vulnerable to invasion, and this may be combined with responses to other environmental cues. Different monogenean species that infect the same host species may adopt different strategies of hatching, indicating that tactics may be more complex than first thought. Control of egg assembly and egg-laying, possibly by host hormones, has permitted colonization of frogs and toads by polystomatid monogeneans. Some monogeneans further improve the chances of infection by attaching eggs to the host or by retaining eggs on, or in, the body of the parasite. The latter adaptation has led ultimately to viviparity in gyrodactylid monogeneans

Invasion of the shovelnose ray (Rhinobatos typus) by Neoheterocotyle rhinobatidis and Merizocotyle icopae (Monogenea : Monocotylidae)

This study examined the route of infection by free-swimming larvae of 2 monocotylid monogeneans that inhabit the gills (Neoheterocotyle rhinobatidis) and the nasal tissue (Merizocotyle icopae) of the shovelnose ray, Rhinobatos typus, from Heron Island on the Great Barrier Reef, Australia. Larvae of N. rhinobatidis and M. icopae attached directly to the gills and the nasal tissue of the ray, respectively, and did not first settle on the skin. Initial development of the post-oncomiracidium of N. rhinobatidis was rapid and hamuli formed between 6 and 24 h p.i. at a mean temperature of 26 °C. However, growth then slowed markedly and was variable; only 2 fully mature individuals were found 20 days p.i. at a mean temperature of 24·5 °C. Development of M. icopae was slow and variable throughout; hamuli did not appear until 10 days p.i. and no mature individuals were obtained even 22 days p.i. at a mean temperature of 24·5 °C. No character could be found as an indicator of parasite age for N. rhinobatidis or M. icopae due to the high variability in development in both species.L. A. Chisholm and I. D. Whittingto

Mechanism of adhesion and detachment at the anterior end of Merizocotyle icopae (Monogenea: Monocotylidae) including ultrastructure of the anterior adhesive matrix

The anterior adhesive mechanism was studied for Merizocotyle icopae (Monogenea: Monocotylidae). Adult anterior apertures can open and close. In addition, duct endings terminating within the apertures are everted or retracted depending on the stage of attachment. Adhesive in adults is synthesized from all 3 secretory types (rod-shaped, small and large spheroidal bodies) found within anterior apertures. All exit together and undergo mixing to produce the adhesive matrix, a process that depletes duct contents. A greater number of ducts carrying rod-shaped bodies is depleted than ducts containing spheroidal bodies which changes the ratio of secretory types present on detachment. Detachment involves elongation of duct endings and secretion of additional matrix as the worm pulls away from the substrate. The change in secretory type ratio putatively modifies the properties of the secreted matrix enabling detachment. Only after detachment do ducts refill. During attachment, individual secretory bodies undergo morphological changes. The larval and adult adhesive matrix differs. Anterior adhesive in oncomiracidia does not show fibres with banding whereas banded fibres comprise a large part of adult adhesive. The data suggest that this is the result of adult spheroidal secretions modifying the way in which the adult adhesive matrix forms.B. W. Cribb, W. D. Armstrong and I. D. Whittingto

Contrasting roles of axonal (pyramidal cell) and dendritic (interneuron) electrical coupling in the generation of neuronal network oscillations

Electrical coupling between pyramidal cell axons, and between interneuron dendrites, have both been described in the hippocampus. What are the functional roles of the two types of coupling? Interneuron gap junctions enhance synchrony of γ oscillations (25-70 Hz) in isolated interneuron networks and also in networks containing both interneurons and principal cells, as shown in mice with a knockout of the neuronal (primarily interneuronal) connexin36. We have recently shown that pharmacological gap junction blockade abolishes kainate-induced γ oscillations in connexin36 knockout mice; without such gap junction blockade, γ oscillations do occur in the knockout mice, albeit at reduced power compared with wild-type mice. As interneuronal dendritic electrical coupling is almost absent in the knockout mice, these pharmacological data indicate a role of axonal electrical coupling in generating the γ oscillations. We construct a network model of an experimental γ oscillation, known to be regulated by both types of electrical coupling. In our model, axonal electrical coupling is required for the γ oscillation to occur at all; interneuron dendritic gap junctions exert a modulatory effect

Evaluation of behavioral changes and subjective distress after exposure to coercive inpatient interventions

BACKGROUND: There is a lack of evidence to underpin decisions on what constitutes the most effective and least restrictive form of coercive intervention when responding to violent behavior. Therefore we compared ratings of effectiveness and subjective distress by 125 inpatients across four types of coercive interventions. METHODS: Effectiveness was assessed through ratings of patient behavior immediately after exposure to a coercive measure and 24 h later. Subjective distress was examined using the Coercion Experience Scale at debriefing. Regression analyses were performed to compare these outcome variables across the four types of coercive interventions. RESULTS: Using univariate statistics, no significant differences in effectiveness and subjective distress were found between the groups, except that patients who were involuntarily medicated experienced significant less isolation during the measure than patients who underwent combined measures. However, when controlling for the effect of demographic and clinical characteristics, significant differences on subjective distress between the groups emerged: involuntary medication was experienced as the least distressing overall and least humiliating, caused less physical adverse effects and less sense of isolation. Combined coercive interventions, regardless of the type, caused significantly more physical adverse effects and feelings of isolation than individual interventions. CONCLUSIONS: In the absence of information on individual patient preferences, involuntary medication may be more justified than seclusion and mechanical restraint as a coercive intervention. Use of multiple interventions requires significant justification given their association with significant distress

Migration site-specificity and development of Benedenia lutjani (Monogenea: Capsalidae) on the surface of its host Lutjanus carponotatus (Pisces: Lutjanidae)

Life-history attributes of the capsalid monogenean Benedenia lutjani, a parasite of Lutjanus carponotatus from the Great Barrier Reef, Queensland, Australia, were investigated from experimental infections. Oncomiracidia of B. lutjani invaded and attached at any site on the fish, but more commonly invaded body surfaces. Immature specimens then migrated to the pelvic fins. Development of the reproductive organs of B. lutjani corresponded with migratory movements on the host. Parasite aggregation on the pelvic fins coincided with the development of functional male reproductive organs and some protandrous worms that possessed a vagina appeared to be inseminated. Migration to, and aggregation on, the branchiostegal membranes (membranous folds posterior to the opercula) coincided with the onset of sexual maturity and commencement of egg production by parasites. The rate of parasite development and the timing of migratory events on the host were influenced by water temperature. All specimens of B. lutjani reached sexual maturity between 12 and 14 days p.i. at 24 °C and between 8 and 10 days p.i. at 27 °C. Anterior hamuli grew continually during a 16-day experiment at 27 °C and 25-day experiment at 23 °C and their length appeared to provide a suitable index to estimate parasite age. The possible adaptive significance of the migratory behaviour, site-specificity and its link with changes in parasite development are discussed.I. D. Whittington and I. Erns

Spatial distribution of mortality in Pacific oysters Crassostrea gigas: reflection on mechanisms of OsHV-1 transmission

The ostreid herpesvirus (OsHV-1) has the potential to devastate Crassostrea gigas culture in Australia as it has done in many other countries, highlighting the need for a better understanding of disease expression and transmission. The aim of this study was to assess the spatial distribution of OsHV-1 associated mortalities in one of only two infected areas in Australia, Woolooware Bay (Botany Bay New South Wales). In October 2011, healthy sentinel Pacific oysters were placed in three different locations at three different tidal levels and OsHV-1 associated mortalities were closely monitored over 7 months. The outbreak started in November 2011 and the disease remained active until April 2012. Three major mortality events were detected. Rather than being a propagating epizootic, it appeared that most oysters were infected from the same environmental source. The distribution of OsHV-1 associated mortalities was spatially clustered, highly variable and clearly dependent on the age of oysters and their position in the water column. Non-random distribution of mortalities at macro scale (sites several km apart) and micro scale (within rearing trays), and vertical clustering patterns in the water column are discussed in relation to mechanisms of transmission in water. We hypothesise that OsHV-1 may be carried through water by particles, possibly plankton. Key words: Crassostrea gigas, Ostreid herpesvirus 1, summer mortalities, spatial distribution, plankton, disease transmissionFunded by the Fisheries Research and Development Corporation, the University of Sydney and the Sydney Metropolitan Catchment Management Authorit

A preliminary phylogenetic analysis of the Capsalidae (Platyhelminthes : Monogenea : Monopisthocotylea) inferred from large subunit rDNA sequences

Phylogenetic relationships within the Capsalidae (Monogenea) were examined using large subunit ribosomal DNA sequences from 17 capsalid species (representing 7 genera, 5 subfamilies), 2 outgroup taxa (Monocotylidae) plus Udonella caligorum (Udonellidae). Trees were constructed using maximum likelihood, minimum evolution and maximum parsimony algorithms. An initial tree, generated from sequences 315 bases long, suggests that Capsalinae, Encotyllabinae, Entobdellinae and Trochopodinae are monophyletic, but that Benedeniinae is paraphyletic. Analyses indicate that Neobenedenia, currently in the Benedeniinae, should perhaps be placed in a separate subfamily. An additional analysis was made which omitted 3 capsalid taxa (for which only short sequences were available) and all outgroup taxa because of alignment difficulties. Sequence length increased to 693 bases and good branch support was achieved. The Benedeniinae was again paraphyletic. Higher-level classification of the Capsalidae, evolution of the Entobdellinae and issues of species identity in Neobenedenia are discussed.I. D. Whittington, M. R. Deveney, J. A. T. Morgan, L. A. Chisholm and R. D. Adlar

Experimental infections of Pacific oyster Crassostrea gigas using the Australian OsHV-1 µVar strain

In Australia the spread of the Ostreid herpesvirus OsHV-1 microvariant (OsHV-1 µVar) threatens the Pacific oyster industry. There was an urgent need to develop an experimental infection model in order to study the pathogenesis of the virus in controlled laboratory conditions. The present study constitutes the first attempt to use archived frozen oysters as a source of inoculum, based on the Australian OsHV-1 µVar strain. Experiments were conducted to test (i) the virus infectivity, (ii) the dose-response relationship for OsHV-1, and (iii) the best storage conditions to keep infective viral inoculum. Intramuscular injection of a viral inoculum consistently led to an onset of mortality 48h post injection and a final cumulative mortality exceeding 90%, in association with high viral loads (1 x 105 – 3 x 107 copies of virus mg-1) in dead individuals. For the first time an infective inoculum was produced from frozen oysters (tissues stored at -80°C for 6 months). Storage of purified viral inoculum at +4°C for 3 months provided similar results to use of fresh inoculum whereas storage at -20°C, -80°C and room temperature was detrimental to infectivity. A dose-response relationship for OsHV-1 was identified but further research is recommended to determine the most appropriate viral concentration for development of infection models that would be used for different purposes. Overall, this work highlights the best practices and potential issues that may occur in the development of a reproducible and transferable infection model for studying the pathogenicity of the Australian OsHV-1 strain in C. gigas under experimental conditions. Keywords: Ostreid herpesvirus 1; Crassostrea gigas; experimental infection model; dose-response; inoculum preparation; storage condition

Egg-laying patterns and in vivo egg production in the monogenean parasites Heteraxine heterocerca and Benedenia seriolae from Japanese yellowtail Seriola quinqueradiata

Egg-laying patterns and egg production in Heteraxine heterocerca from the gills and Benedenia seriolae from the skin of Japanese yellowtail Seriola quinqueradiata in Japan were investigated in vivo. Eggs were collected every 3 h from sexually mature H. heterocerca and B. seriolae infecting 3 S. quinqueradiata kept individually over 3 consecutive days and exposed to alternating periods of illumination and darkness (LD 12:12; light on 06.00, light off 18.00) and maintained at 23.8 +/- 0.1 degrees C and 35 ppt salinity. A well-defined egg-laying rhythm was demonstrated for H. heterocerca while B. seriolae was shown torelease eggs continuously. A total of 114,000 H. heterocerca eggs was collected and of these, 45.4 (42.5-49.9)% were collected during the first 3 h period following dark at 18.00 h. A total of 662,857 B. seriolae eggs was collected and these were distributed over each 3 h period ranging from 11.1 to 14.1% of the daily egg output. All eggs extracted from the uterus of each H. heterocerca were joined together forming an 'egg-string'. The contrasting egg-laying patterns of H. heterocerca and B. seriolae suggest that each species makes use of a different infection strategy to infect the same host species, S. quinqueradiata.A. J. Mooney, I. Ernst and I. D. Whittingto