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• W1566199991 abstract "Rock lobster hatchery rearing for the supply of juvenile seedstock to aquaculture has yet to becommercialized due to the complex and prolonged larval phase, which is further confoundedby bacteria-associated infections and mortalities. However, the adoption of improved larvalrearing technology and the practice of hygienic husbandry at the Tasmanian Aquaculture andFisheries Institute Marine Research Laboratories (TAFI MRL) have been responsible forclosing the life cycle of the southern rock lobster Jasus edwardsii and more recently of theeastern rock lobster Jasus (Sagmariasus) verreauxi. The aim of this research was to control themicroorganisms that colonize the phyllosoma during their early stages of development,especially by application of bacteria with probiotic characteristics. Bacteria were isolated andidentified from the microbial community of phyllosoma cultured at TAFI MRL and wereexamined for their probiotic or pathogenic effects.Since pathogenic, opportunistic, beneficial and innocuous Vibrio spp. are ubiquitous incrustacean hatcheries and due to the prevalence of Vibrio spp. in clear water larval rearingtechniques, the probability of obtaining autochthonous Vibrio spp. probionts that have bettercolonizing efficacy than probionts that are used in terrestrial animals was considered to be high.Bacterial isolates from live and dead J. edwardsii phyllosoma (instar I, n = 100; instars III andVI, n = 50) were randomly selected from bacteria enumerated on Johnson's marine agar andTCBS agar plates, and subculturing produced 239 viable isolates from 25 different species. Theisolates were identified using PIBWIN based on the biochemical reactions conducted with theMicroSys® Vibrio identification kit. Different species that constituted the microbial communityof the phyllosoma were designated as putative probionts or likely pathogens based on theirassociation with live and dead phyllosoma, respectively. High recovery of V. chagasii from dead instar I phyllosoma and its complete absence from live instar I phyllosoma indicated thatit was likely to be pathogenic. Similarly, high occurrence of V. splendidus from dead instar IIIand VI phyllosoma indicated it to be a virulent isolate. Following this hypothesis, V.mediterranei, V. alginolyticus, Phenon 36, Phenon 52, V. cyclitrophicus, and V. calviensis werealso presumed to be pathogenic. Phenon 8, V. orientalis, V. anguillarum and V. penaecida wereassociated with healthy, live phyllosoma. Isolates that were identified as Phenon' were V.alginolyticus-like isolates.The development of a probiotic model to protect phyllosoma was an important aspect of thisresearch. Since hatchery-reared phyllosoma derive their gut microflora from Artemia, thedelivery of isolates via Artemia during the present study mimicked the normal route throughwhich the bacterially-free phyllosoma become colonized. The Artemia were subjected to ashort disinfective purge (with formaldehyde and concentrated microalgae) and immediatelysuspended in microalgae inoculated with the axenic bacterial isolates. The bacterially colonizedArtemia were dispensed into the phyllosoma rearing containers and the gut evacuation of theisolates by the Artemia was determined. During enrichment, Artemia accumulated 1.3-4.9 ±0.02-0.1 x 107 and 1.2-2.5 ± 0.01-0.07 x 107 heterotrophic and Vibrio cells Artemia-1 ,respectively, and 24 h later, the numbers had diminished 100-fold. Experiments determiningthe effect of monoxenic isolates on the survival of newly hatched phyllosoma were conductedwith isolates that were often or exclusively obtained from either dead or live phyllosoma.During these experiments, which lasted for 14 days, V. penaecida (Sr. No. 232) and V. chagasii(Sr. No. 64) were pathogenic towards phyllosoma with 22.5% and 58% survival, respectively,compared to 68% survival in the Control. The survival in phyllosoma exposed to monoxeniccultures of Phenon 8 (Sr. No.148), Phenon 52 (Sr. No. 14), V. cychtrophicus (Sr. No. 152) and V. orientalis (Sr. No. 229) was 67-87%. V. mediterranei (Sr. No. 93), V. calviensis (Sr. No. 30),V. alginolyticus (Sr. No. 25), V. splendidus (Sr. No. 166), V. anguillarum (Sr. No. 221) andPhenon 36 (Sr. No. 215) were mildly virulent towards phyllosoma (61-65% survival) comparedto the Control. The histological sections of dead phyllosoma revealed proliferation of bacterialcells in the lumen.Probiont-pathogen challenge experiments were conducted on phyllosoma, wherein V.cyclitrophicus (Sr. No. 152), V. orientalis (Sr. No. 229), Phenon 8 (Sr. No.148) and Phenon 52(Sr. No. 14) were used as putative probionts while V. penaecida (Sr. No. 232) and V. chagasii(Sr. No. 64) were used as pathogens. Probiont-pathogen treatments received putative probiontsexclusively until Day 4 and thereafter also received the pathogens until the experiments wereterminated. Survivals in the probiont-pathogen treatments were significantly higher than in thetreatments receiving pathogen-only, and were not statistically different from survivals in theControl or the probiont-only treatments. Highest survival (88%) was in the Phenon 8 (Sr.No.148) - V. chagasii (Sr. No. 64) treatment. Other probiont-pathogen treatments had 86-82%survival with the exception of V. orientalis (Sr. No. 229) — V. penaecida (Sr. No. 232), whichhad 75% survival. Probiotic effects are exerted through mechanisms such as in vivoantagonism, competitive exclusion of the pathogens by the probionts, activation of the innatecrustacean immune responses and production of siderophores. During the present study, theobserved probiotic effect may have involved either a single mechanism or a combination ofmechanisms of probiosis. High phyllosoma survival was associated with high abundance ofsucrose fermentors while sucrose non-fermentors typically accompanied poor survival. The therapeutic effect of the probionts was determined by administering the probionts tophyllosoma that had been previously colonized by V. chagasii (Sr. No. 64) and V. penaecida(Sr. No. 232). The administration of probionts was able to halt the phyllosoma mortality andgradually displaced the pathogen. The administration of the probionts and pathogensimultaneously to phyllosoma infected with the pathogens improved the survival compared tothe treatments that received the pathogen exclusively throughout the experiment. There was nobenefit of mixed-culture probionts over axenic probionts on the survival of phyllosoma.Further, the application of probionts and intermittent ozonation of the culture water did notimprove larval survival, although the effect of ozonation resulted in a reduction in the bacterialabundances.The application of autochthonous probionts as prophylactic and therapeutic measures ensuredhigh survival of phyllosoma during their critical early larval phase. The results of this studymay assist in manipulating and controlling the bacterial colonization of early stage phyllosomaduring hatchery rearing." @default.
• W1566199991 created "2016-06-24" @default.
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• W1566199991 date "2009-01-01" @default.
• W1566199991 modified "2023-09-24" @default.
• W1566199991 title "Probiotic and pathogenic bacteria in larval rearing of spiny lobsters, Jasus edwardsii and Jasus (Sagmariasus) verreauxi" @default.
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