Chemical changes in fed and starved larval trout cod, Maccullochella macquarensis during early development

The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in μmoles larva^–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 μg mg lipid^–1 larva^–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 μg mg lipid^–1 larva^–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.

Phylogenetic relationships of the globally distributed freshwater prawn genus Macrobrachium (Crustacea: Decapoda: Palaemonidae): biogeography, taxonomy, and the convergent evolution of abbreviated larval development

There has hitherto been little research into evolutionary and taxonomic relationships amongst species of the freshwater prawn genus Macrobrachium Bate across its global distribution. Previous work by the authors demonstrated that the endemic Australian species did not evolve from a single ancestral lineage. To examine whether other regional Macrobrachium faunas also reflect this pattern of multiple origins, the phylogeny of 30 Macrobrachium species from Asia, Central/South America and Australia was inferred from mitochondrial 16S rRNA sequences. Phylogenetic relationships demonstrate that, despite some evidence for regional diversification, Australia, Asia and South America clearly contain Macrobrachium species that do not share a common ancestry, suggesting that large-scale dispersal has been a major feature of the evolutionary history of the genus. The evolution of abbreviated larval development (ALD), associated with the transition from an estuarine into a purely freshwater lifecycle, was also mapped onto the phylogeny and was shown to be a relatively homoplasious trait and not taxonomically informative. Other taxonomic issues, as well as the evolutionary origins of Macrobrachium, are also discussed.

Induced spawning, larval development and rearing of two indigenous Malaysian mahseer, Tor tambroides and T. douronensis

The Mahseers (Tor spp.) are highly valued freshwater fishes across the Himalayan and South-east Asian regions. Over exploitation of natural stocks because of high demand and the deteriorating environmental conditions have resulted in marked decline of mahseers in the wild. Malaysian mahseers, T. tambroides (Bleeker) and T. douronensis (Valenciennes), locally known as empurau, kelah or belian and semah, respectively, have significant cultural and economic importance but both species are now threatened in the wild because of environmental degradation and over fishing. A captive breeding programme was instigated to attempt to propagate these two species artificially for conservation and aquaculture purposes. Both pond-reared and tank-held T. tambroides and T. douronensis reached sexual maturity in captivity and were successfully induced to spawn using hormone treatments. Ovaprim (0.5 mL kg⁻¹) was the most successful hormone treatment for both species. Pre-treatment of fish with Ovaplant (28–68 μg kg⁻¹, 2–7 weeks before spawning induction) greatly improved the success rate of spawning induction. Repeat spawning (within 4 months of initial spawning) was induced in some captive fish. Use of formalin baths improved hatching by preventing fungal infections. Embryonic development and hatching are described. Juveniles were reared in static greenwater ponds. Tor tambroides reached 142–179 g (max 270 g) in 60 weeks. These results represent the first successful captive spawning and rearing of both species. Options for future research to improve production are discussed.

Culture-based fisheries in non-perennial reservoirs of Sri Lanka: influence of reservoir morphometry and stocking density on yield

Culture-based fish yield in non-perennial reservoirs of Sri Lanka was related to reservoir morphometry and stocking density. The reservoirs were stocked mainly with fingerlings of one Chinese and three Indian major carp species, common carp, Cyprinus carpio L., and the genetically improved farmed tilapia strain of Nile tilapia, Oreochromis niloticus (L.), at four pre-determined species combinations and a range of stocking densities [SD (fingerlings ha⁻¹)]. Twenty-three reservoirs were harvested successfully at the end of the culture period of 2002–2003. Basic limnological and morphometric parameters, including shoreline development (D_L) and shoreline area ratio (R_LA), were estimated for each of the 23 reservoirs. Bray–Curtis similarity and non-metric multidimensional scaling using mean values of limnological data revealed that reservoirs could be ordinated into two major clusters, one with intact sample distribution due to similar trophic characteristics and the other with scattered sample distribution. Reservoirs in the cluster with similar trophic characteristics showed significant correlation (P < 0.05) between R_LA and total fish yield (Y). A multiple regression equation, Y = −693 + 4810 R_LA + 0.484 SD, was generated to estimate fish harvest in relation to SD.

The presence of GABA in gastropod mucus and its role in inducing larval settlement

Chemical substances that induce larval settlement have been the focus of many gastropod studies due to the importance of wild stock recruitment and production within aquaculture facilities. Gamma-aminobutyric acid (GABA), GABA analogs, and GABA-mimetics associated with certain crustose coralline algae (CCA), are known to induce larval settlement in commercial abalone (Haliotis) species, and other gastropods. Furthermore, mucus secreted from these gastropods has been shown to induce larval settlement, but the stimulatory components of mucus have not been thoroughly investigated. We now present data confirming that GABA is the settlement-inducing effector molecule contained within abalone mucus. To do this, we initially generated anti-GABA for use in immunoenzyme and immunofluorescent microscopy. Using these techniques GABA was identified in the nerves and epithelial cells of the foot, including mucus. Dried mucus samples subject to HPLC analysis revealed a mean concentration of 0.68 mM GABA after sample rehydration. The presence of GABA in these samples was confirmed by time-of-flight mass spectroscopy (TOF-MS). In addition, GABA was detected in the mucus of several abalone species and other gastropods by immunocytochemistry. Subsequent bioassays using both dry and fresh mucus strongly promoted induction of larval settlement.

Larval settlement of the tropical abalone, Haliotis asinina Linnaeus, using natural and artificial chemical inducers

Many kinds of chemical and biological materials have been used as inducers of settlement of abalone larvae, as well as other species of marine gastropods, with responses being highly variable, even to the same chemical cue. The present study tested chemical inducers, γ-aminobutyric acid (GABA), δ-aminovaleric acid (5-AVA) and l-glutamic acid (GA) and the effects they have on larval settlement of Haliotis asinina. Additionally, a relatively inexpensive commercial substance, monosodium glutamate (MSG), was trialed. The datum provided shows all chemicals to be active inducers of settlement in this study, in order of effectiveness of 5-AVA, GABA, MSG to GA. Induction as adjudged from larval numbers settled was best at 6 h 62%, with 10⁻¹ mM 5-AVA. At 24 h, induction was the highest at 78% when exposed to 10⁻² mM 5-AVA. Larvae that were allowed to settle up to 72 h showed the highest numbers of settled larvae, and declined back to 60% when exposed to 10⁻² 5-AVA and 10⁻¹ mM GABA respectively. Monosodium glutamate, although third in settlement standings would bypass the other chemicals, with regard to cost versus yield. The assessment of settlement surface, rough or smooth proved to be irrelevant, which had no significant impact on larval settlement.

Invertebrate larval dynamics in seasonally closed estuaries

Estuarine benthic assemblages are often numerically dominated by polychaetes. The limits of these populations are determined by larval, and probably to a lesser extent adult movement. A previous study (Newton 1996), indicated that planktonic polychaete larvae were very abundant over the summer months in the Hopkins River; however, the identification and source of these larvae was not known. Defining the extent of a population, and therefore the likelihood of that population recovering following a perturbation, is crucial for effective estuarine management. This study investigated both the likely source of the larvae, (i.e. estuarine or marine) and the extent of larval dispersal within and between estuaries by addressing the following questions: Which taxa produced the planktonic larvae? Are these taxa resident estuarine species? Are the larvae of different taxa evenly distributed within the estuary or do physicochemical parameters or other factors influence their abundance? Are the same larvae found in other estuaries along the coast? and Is there exchange of these larval taxa with the marine environment and other estuaries? Larvae were identified and described by culturing commonly occurring planktonic larvae until adult characteristics appeared. The spionids, Carazziella victoriensis and Prionospio Tatura, numerically dominated the plankton in the Hopkins and the spionid, Orthoprionospio cirriformia was recorded from the Hopkins, Curdies and Gellibrand estuaries. Two spionids, Carazziella sp. and Polydora sp. were identified from tidal waters. Mouth status and physicochemical conditions (salinity, temperature and dissolved oxygen) were monitored in each estuary. Whereas the Merri and Gellibrand estuaries were predominantly stratified over the sampling period, the Curdies was more often well mixed and the Hopkins varied from well mixed to stratified. The duration of mouth opening and hence the opportunity for larval exchange also varied in each estuary. The Merri River was closed for 13.5% of days over the study period, the Gellibrand River for 18.4%, the Hopkins River for 49% and the Curdies River for 71.0%. The distributions of larvae at spatial scales of metres, 100s of metres and kilometres were investigated within a single estuary. While the same larvae, C. victoriensis, P. Tatura and bivalve larvae, were found along the length of the Hopkins estuary the abundances varied at different spatial scales suggesting different processes were influencing the distribution of P. Tatura larvae, and C. victoriensis and bivalve larvae. The distribution of larvae between several estuaries was investigated by monitoring meroplankton at two sites at the mouth of each of the four estuaries approximately monthly (except for winter months). Different meroplanktonic assemblages were found to distinguish each estuary. Further, C. victoriensis and P. Tatura larvae were only recorded in the Hopkins but larvae of the spionid, Orthoprionopio cirriformia were detected in the Hopkins, Curdies and Gellibrand estuaries. The extent of larval exchange with other estuaries and the marine environment was determined by monitoring tidal waters. Settlement trays were also deployed to determine if larvae were moving into estuaries and settling but not recruiting. P. tatura larvae were not detected in the tidal waters of any estuary and while C. victoriensis and O. cirriformia were found in both flood and ebb tides there was no evidence of movement of theses taxa to other estuaries. Larvae of the spionids, Carazziella sp. and Polydora sp., were found in tidal waters of each estuary but were rarely detected in the plankton within the estuaries. Neither species was found as an adult in background cores from any estuary, nor with the exception of a few individuals in the Merri, were they detected in settlement trays in any estuary. I conclude that the source of the larvae of C. victoriensis, P. Tatura and O. cirriformia is estuarine and while C. victoriensis, and O. cirriformia move in and outh of the source estuary in tidal waters there was no evidence for movement to other estuaries. The spionids, Carazziella sp. and Polydora sp were considered to be marine and while they moved in and out of estuaries in tidal waters they did not usually settle in the estuaries. The results of this study are a crucial first step in the development of ecological models to better understand dispersal in seasonally closed estuaries that are typical of southern Australia. This study emphasises the unique physicochemical characteristics and biological assemblages within these estuaries and the need for estuarine management to reflect these differences.

Nutritional related aspects of the artificial propagation and larval rearing of Macquarie perch, Macquaria Australasica (Cuvier, 1830)

Studies the underlying role of nutrition in the lack of response of captive fish to hypophysation. Aspects studied include morphological characteristics, histology of ovaries, proximate analysis, fatty and amino acid profiles of oocytes, muscle, liver and diets of wild and tank-reared fish, egg and larval quality, amino acid composition of eggs and larvae at different developmental stages, larval feeding and hormone treatments.

Reproductive performance and egg and larval quality of the blacklip abalone

The study highlights the interrelationship between physical characteristics of female broodstock with reproductive performance, egg quality and subsequent larval success, and reproductive seasonality of the blacklip abalone (Haliotis rubra Leach). Suitable indices for assessing broodstock were developed. Seasonal changes in biochemical compositions in relation to maturation and development was discussed.

Heat tolerance, behavioural temperature selection and temperature-dependent respiration in larval Octopus huttoni

This study reports temperature effects on paralarvae from a benthic octopus species, Octopus huttoni, found throughout New Zealand and temperate Australia. We quantified the thermal tolerance, thermal preference and temperature-dependent respiration rates in 1-5 days old paralarvae. Thermal stress (1°C increase h^-1) and thermal selection (~10-24°C vertical gradient) experiments were conducted with paralarvae reared for 4 days at 16°C. In addition, measurement of oxygen consumption at 10, 15, 20 and 25°C was made for paralarvae aged 1, 4 and 5 days using microrespirometry. Onset of spasms, rigour (CT_max) and mortality (upper lethal limit) occurred for 50% of experimental animals at, respectively, 26.0±0.2°C, 27.8±0.2°C and 31.4±0.1°C. The upper, 23.1±0.2°C, and lower, 15.0±1.7°C, temperatures actively avoided by paralarvae correspond with the temperature range over which normal behaviours were observed in the thermal stress experiments. Over the temperature range of 10°C-25°C, respiration rates, standardized for an individual larva, increased with age, from 54.0 to 165.2nmol larvae^-1h^-1 in one-day old larvae to 40.1-99.4nmol h^-1 at five days. Older larvae showed a lesser response to increased temperature: the effect of increasing temperature from 20 to 25°C (Q₁₀) on 5 days old larvae (Q₁₀=1.35) was lower when compared with the 1 day old larvae (Q₁₀=1.68). The lower Q₁₀ in older larvae may reflect age-related changes in metabolic processes or a greater scope of older larvae to respond to thermal stress such as by reducing activity. Collectively, our data indicate that temperatures >25°C may be a critical temperature. Further studies on the population-level variation in thermal tolerance in this species are warranted to predict how continued increases in ocean temperature will limit O. huttoni at early larval stages across the range of this species.

Effects of bait age, larval chemical cues and nutrient depletion on colonization by forensically important calliphorid and sarcophagid flies

Species colonization patterns on corpses and the frequency of carrion fly oviposition and larviposition are affected by decomposition stage and previous maggot colonization. This study investigated these effects on meat bait colonization by Victorian Diptera of forensic importance. Bait treatments were: 'aged' (aged for 4 days at 22 °C, allowing some decomposition); 'nutrient-depleted' [aged for 4 days at 22 °C with feeding Calliphora vicina (Robineau-Desvoidy) (Diptera: Calliphoridae) larvae]; 'extract' (fresh bait mixed with liquid formed by feeding C. vicina larvae), and 'fresh' (untreated control bait). Statistical analysis (α = 0.05) revealed that colonization frequency differed significantly among treatments (Welch's F 3,18.83 = 4.66, P < 0.05). Post hoc tests showed that fresh and extract baits were colonized extensively throughout the experiment with no significant difference, whereas the colonization of nutrient-depleted baits was significantly lower. This suggests that larval digestive enzymes, larval excreta and cuticular hydrocarbons have less effect on colonizing Diptera than the nutritional content of meat. The colonization of aged baits did not differ significantly from that of fresh, extract or nutrient-depleted baits. A further experiment testing 'very aged' (aged for 8 days at 28 °C), 'larvae-added' (fresh bait with C. vicina larvae added before placement) and 'fresh' (untreated control) baits revealed that very aged baits were colonized significantly less frequently than either fresh or larvae-added baits (Welch's F 2, 6.17 = 17.40, P < 0.05).

Larval development of himri, Barbus luteus (Cyprinidae: Cypriniformes) reared in the laboratory

Larval development of the himri barbel, Barbus luteus Heckel, reared at 19-22 ¡C in natural daylight is described. Morphological and some functional character appearances were similar to the main ontogenetic steps of development in most cyprinids. Small hatched larvae attained both metamorphosis and a length of 10.5 mm after almost 35 days. Relative growth of the selected body proportions demonstrated typical priorities in growth compared to the length. An ontogenetic index was inferred from applying age and length criteria at metamorphosis as a scale for ontogenetic events.