Histological alterations in the Antarctic cod, Trematomus bernacchii from Davis Station, East Antarctica

During the summer season 2009/10, a comprehensive environmental impact assessment of the sewage discharge was conducted at Davis Station in the Vestfold Hills region of East Antarctica. As part of this project, a survey of the histology of liver, gill, gonad and muscle tissues in the Antarctic rock cod Trematomus bernacchii from nearshore sites in the receiving environment close to Davis Station in was completed. Fish from 4 sites were examined; 1 site adjacent to the Davis Station sewage outfall (within 500 m of the point of discharge), 2 sites approximately 2 km from the outfall (Anchorage Island and Antennae farm), and 1 site approximately 10 km away from the outfall and adjacent to an Adelie penguin population (Kazak Island). All fish sampled from the sewage outfall site exhibited significant histological alterations in all major tissues. Fish from the other 3 sites showed some alterations in either gill and/or liver tissues. Pathological abnormalities present in all fish collected near the sewage outfall included: extensive multifocal cysts of unknown etiology with necrotic liquification; multifocal granuloma with associated inflammation; coagulative necrosis in the liver; and lamellar hyperplasia with associated proliferation and lamella fusion of the gills. Results of this work form part of a weight of evidence approach alongside ecological monitoring, chemical analysis, ecotoxicological testing and dispersal modelling of the discharge plume which is being used to inform and direct upgrades to the Australian Antarctic Divisions operations and current sewage discharge practises at Davis Station.

Application of a quantitative histological health index for Antarctic rock cod (Trematomus bernacchii) from Davis Station, East Antarctica

A quantitative Histological Health Index (HHI) was applied to Antarctic rock cod (Trematomus bernacchii) using gill, liver, spleen, kidney and gonad to assess the impact of wastewater effluent from Davis Station, East Antarctica. A total of 120 fish were collected from 6 sites in the Prydz Bay region of East Antarctica at varying distances from the wastewater outfall. The HHI revealed a greater severity of alteration in fish at the wastewater outfall, which decreased stepwise with distance. Gill and liver displayed the greatest severity of alteration in fish occurring in close proximity to the wastewater outfall, showing severe and pronounced alteration respectively. Findings of the HHI add to a growing weight of evidence indicating that the current level of wastewater treatment at Davis Station is insufficient to prevent impact to the surrounding environment. The HHI for T. bernacchii developed in this study is recommended as a useful risk assessment tool for assessing in situ, sub-lethal impacts from station-derived contamination in coastal regions throughout Antarctica.

Direct evidence of histopathological impacts of wastewater discharge on resident Antarctic fish (Trematomus bernacchii) at Davis Station, East Antarctica.

During the 2009/2010 summer, a comprehensive environmental impact assessment (EIA) of the wastewater discharge at Davis Station, East Antarctica was completed. As part of this, histological alteration of gill and liver tissue in Antarctic Rock-cod (Trematomus bernacchii) from four sites along a spatial gradient from the wastewater outfall were assessed. All fish within 800 m of the outfall exhibited significant histological changes in both tissues. Common pathologies observed in fish closest to the outfall include proliferation of epithelial cells with associated secondary lamellar fusion in the gills and multifocal granulomata with inflammation and necrosis as well as cysts in the liver. Fish from sites >800 m from the outfall also exhibited alterations but to a lesser degree, with prevalence and severity decreasing with increasing distance from the outfall. This study highlights the value of histopathological investigations as part of EIAs and provides the first evidence of sub-lethal alteration associated with wastewater discharge in East Antarctica.

Performance of juvenile Murray cod, Maccullochella peelii peelii (Mitchell), fed with diets of different protein to energy ratio

The results of a 56-day experiment on juvenile Murray cod, Maccullochella peelii peelii, an Australian native fish with a high aquaculture potential, of mean weight 14.9 ± 0.04 g, fed with five experimental diets, one a series of 40% protein content and lipid levels of 10, 17 and 24% (P40L10, P40L17 and P40L24), and another of 50% protein and 17 and 24% (P50L17 and P50L24) lipid are presented. The specific growth rate (SGR) (% day−1) of fish maintained on different diets ranged from 1.18 to 1.41, and was not significantly different between dietary treatments, except P40L10 and the rest. However, there was a general tendency for SGR to increase with increasing dietary lipid content at both protein levels. The food conversion ratio (FCR) for the 40% protein series diets were poorer compared with those of the 50% protein diets, and the best FCR of 1.14 was observed with the P50L17 diet. The protein efficiency ratio (PER), however, was better in fish reared on low protein diets. The net protein utilization (NPU) also did not differ significantly (P > 0.05) in relation to dietary treatment. As in the case of PER the highest NPU was observed in Murray cod reared on diet P40L24 and the lowest in fish fed with diet P50L24. The carcass lipid content reflected that of the diets, when significant increases in the lipid content was observed in relation to dietary lipid content at both protein levels. However, body muscle lipid content did not increase with increasing dietary lipid content, and was significantly lower than in the whole body. The fatty acids found in highest concentration amongst the saturates, monoenes and polyunsaturates (PUFAs) were 16 : 0, 18 : 1n-9 and 22 : 6n-3, respectively, and each of these accounted for more than 60% of each of the group's total. The muscle fatty acid content was affected by the dietary lipid content; for example the total amount (in μg mg−1 lipid) of monoenes ranged from 72 ± 5.1 (P40L10) to 112 ± 10 (P40L24) and 112 ± 2.8 (P50L17) to 132 ± 11.8 (P50L24) and the n-6 series fatty acids increased with increasing dietary lipid content, although not always significant. Most notably, 18 : 2n-6 increased with the dietary lipid level in both series of diets.

Growth and nutrient utilization of Murray cod Maccullochella peelii peelii (Mitchell) fingerlings fed diets with varying levels of soybean meal and blood meal

The Australian native freshwater fish Murray cod, Maccullochella peelii pellii (Mitchell), currently supports a fledgling inland aquaculture industry, which is thought to have considerable growth potential. The aim of this study was to evaluate the suitability of two alternate protein sources [blood meal (BM) and defatted soybean meal (SBM)] as substitutes for fish meal at various levels of inclusion in diets for juvenile Murray cod. The growth performance of juvenile Murray cod in response to nine isonitrogenous and isocalorific diets (50% protein, 14% lipid, 20.2 kJ g⁻¹) consisting of a control diet in which protein was supplied from fish meal, and test diets in which the fish meal protein was substituted at levels of 8%, 16%, 24%, and 32% with BM or SBM was evaluated from a 70-day growth experiment. The per cent apparent dry matter (% ADC_dm) and percentage protein digestibility (% ADC_p) of the test diets were also determined using Cr₂O₃ as a marker. Survival in all the SBM dietary treatments was high but that of fish on the BM dietary treatments was significantly (P < 0.05) lower than in all the other dietary treatments. Specific growth rate (% day−1) of Murray cod fed SBM incorporated diets ranged from 1.63 ± 0.06 to 1.78 ± 0.10 and even at the highest level tested (32% of the dietary protein from SBM) was not significantly different (P > 0.05) from the fish fed the control diet (1.65 ± 0.09). Feed conversion ratios of the SBM dietary treatments ranged from 1.36 ± 0.08 to 1.45 ± 0.07. The protein efficiency ratios and protein conversion efficiencies of Murray cod in the soybean meal treatments were also good and for a majority of the SBM diets were better than those for the control diet. Per cent ADC_dm and ADC_p of the SBM diets tested ranged from 70.6 ± 1.46 to 72.3 ± 1.81% and 88.6 ± 0.57 to 90.3 ± 0.17%, respectively, and was not significantly different (P > 0.05) from the control diet (% ADC_dm 74.3 ± 1.63; % ADC_p 91.3 ± 0.55). The reasons for significantly poor survival and growth of Murray cod reared on BM incorporated diets, and relatively poor digestibility of these diets are discussed. The study shows that for Murray cod diets in which fish meal protein is substituted up to 32% performance or carcass composition is not compromised.

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.

Microsatellite loci for studies of wild and hatchery Australian Murray cod Maccullochella peelii peelii (Percichthyidae)

Murray cod is a species of conservation concern that is subject both to wild harvest and to aquaculture production. Six polymorphic microsatellite loci have been developed for this species, which will facilitate studies of wild-stock structure, will help the management of hatchery diversity, and will aid genetic improvement programmes. The markers exhibited nonindependence of genotypes between loci and deviations from Hardy–Weinberg expectations, although these most probably reflect practices at the hatchery from which the genotyped individuals were sourced.

Performance of intensively farmed Murray cod Maccullochella peelii peelii (Mitchell) fed newly formulated vs. currently used commercial diets, and a comparison of fillet composition of farmed and wild fish

Murray cod is a top-order carnivore with high culture potential. Currently, there are no commercial diets formulated specifically for Murray cod. In this study, results of two growth trials on Murray cod (80–83.5-g mean initial weight), conducted in commercial settings, using two laboratory-formulated diets (DU1 and DU2; 48.9% and 49.1% protein, and 16.9% and 16.1% lipid, respectively, on a dry matter basis), and two commercial diets, formulated for other species (salmon – CD/S and barramundi – CD/B) but used in Murray cod farming are presented. The two commercial diets had less protein (46.6% and 44.4%) but higher lipid (21.7% and 19.5%). The energy content of the feeds tested was similar (about 20–22 kJ g−1). The growth performance and feed utilization of Murray cod did not differ significantly amongst the diets, but the food conversion ratio and % protein efficiency ratio in fish fed the DU1 and DU2 diets were consistently better. There was significantly less carcass and muscle lipid deposition in fish fed with the latter diets. Of the fatty acids in muscle, the lowest amounts (in μg mg lipid−1) of n-3 (262.5±2.9), n-6 (39.8±0.9) and polyunsaturated fatty acid (PUFA) (302.3±3.8) were observed in fish fed CD/S, and the highest in fish fed DU2 and CD/B. Fatty acids 16:0 and 18:0, 18:1n-9 and 16:1n-7, and 22:6n-3, 20:5n-3, 22:5n-3 and 18:2n-6 were the dominant fatty acids amongst the saturates, monoenes and PUFA, respectively, and accounted for 80.8–88.7% of all identified fatty acids (23) in muscle of Murray cod. The study showed that Murray cod could be cultured successfully on a diet (DU2) containing 20% soybean meal without compromising growth and/or carcass quality. Differences in the proximate composition and fatty acid composition of muscle of wild and farmed Murray cod were observed, the most obvious being in the latter. Wild Murray cod had significantly less (P<0.05) saturates (192.6±1.84 vs. 266.3±3.51), monoenes (156.5±8.7 vs. 207.6±6.19), n-3 (145.2±5.24 vs. 261.8±3.2) but higher n-6 (144.3±2.73 vs. 48.3±1.38) in muscle (all values are in μg mg lipid−1) than in farmed fish. Wild fish also had a much lower n-3 to n-6 ratio (1.0±0.03 vs. 5.4±0.09).

Seasonal variability in whale encounters in the Western Antarctic Peninsula

Cetacean sighting surveys were conducted as part of nine multidisciplinary research cruises over late summer, autumn and winter of 2 years (2001–2003) during the Southern Ocean Global Ocean Ecosystems (SO GLOBEC) program. Sea-ice cover differed markedly between years, with apparent effects on cetacean distribution. No ice was present until late June in 2001, while the previous winter sea ice never fully retreated (>30% cover) during the 2002 or 2003 summer, thus increasing the proportion of thicker and more complex ice, including multi-year floes. Humpback (237 sightings; 537 individuals) and minke (103 sightings: 267 individuals) whales were the most commonly detected species. Data from seven comparable cruises were used to identify habitat for minke and humpback whales over five geographically distinct spatial divisions in the study area. In all years, both species were predominantly found in near coastal habitat, particularly in the fjords where complex habitat likely concentrated prey. In 2002 and 2003 the presence of sea ice provided additional feeding habitat, and the numbers of minkes (in winter) and humpbacks (late summer and autumn) in the area doubled compared with 2001. Humpbacks in particular were concentrated at the ice boundaries during late summer and autumn, while minke numbers increased in the winter that followed and occupied ice-covered areas along the entire shelf edge. Important resource sites for these species are mainly located in near-coastal areas and are used in all years, but when ice margins exist and intersect with resource sites they attract much larger numbers of animals due to the dynamics between sea ice and prey.

Description of loxocythere (novoloxocythere) pelius subgen. sp. nov. (ostracoda) from the cenozoic of S.E. Australia with comments on species of antarctiloxoconcha hartmann, 1986 and loxoreticulatum benson, 1964 from Australian and Antarctic marine waters

Loxocythere (Novoloxocythere) pelius subgen. et sp. nov. is described from Upper Miocene strata of the Port Phillip and Western Port Basins. Victoria. It has its acme in shallow open marine facies of latest Miocene (Cheltenhamian) age. This species, along with Loxocythere (Novoloxocythere) kerryswansoni Yassini and Jones, 1995, forms a discrete group of rotund Australian Loxocythere species that possess posterior extremities in both valves that are positioned well above mid carapace height (i.e. adjacent to dorsal margin). This feature along with a sub-triangular inner margin outline, defines a carapace shape that is distinct from that of rotund species of Loxocythere (loxocythere) Hornibrook, 1952 and Antarctiloxoconcha Hartmann, 1986. The type species of Antarctiloxoconcha – A.frigida (Neale. 1967), possesses internal carapace features that are very similar to the type species of Loxocythere - L. crassa Hornibrook, 1952. Both have relatively short carapaces and sub-quadrate inner margin outlines with posterior extremities in both valves positioned below mid carapace height. Species of Loxocythere (Novoloxocythere), in particular L. (N.) kerryswansoni, have a carapace shape that is transitional between Loxocythere and Loxoreticulatum Benson, 1964. Species of Loxoreticulatum generally possess a sub-parallelogram shaped carapace/inner margin and arched median hinge element. The latter feature is distinct from the mostly straight median hinge elements of Loxocythere (Loxocythere) and Loxocythere (Novoloxocythere) species. Species of Loxocythere (Novoloxocythere) are also readily distinguishable from relatively elongate species of Loxocythere, such as L. (L.) hornibrooki McKenzie, 1967, as the latter possess long (for genus), sub-rectangular shaped carapaces/inner margin outlines and posterior extremities below mid height.

Performance of murray cod, Maccullochella peelii peelii (Mitchell) in response to different feeding schedules

The Australian freshwater fish Murray cod, Maccullochella peelii peelii (Mitchell) is gaining popularity as a suitable species for intensive culture, particularly in closed systems. The aim of this study was to evaluate the performance of Murray cod in response to different feeding schedules. Growth, survival, food conversion and a range of other related parameters including carcass proximate composition were evaluated for fish in five feed management regimes. The feeding regimes used in the experiment were hand fed to satiation twice daily (SAT), a pre-determined ration of 1.2% of the body weight day−1 which was hand fed twice daily (HFR), and belt fed through the day only (B/D), belt fed through the night only (B/N) and belt fed for 24 h (B/DN). Each of the five feeding regimes was randomly allocated to three tanks (triplicates). All of the feeding regimes used a commercially prepared diet formulated specifically for Murray cod, containing ≈50% protein and ≈16% lipid. The experiment was conducted for 84 days. Specific growth rate ranged from 0.89±0.01 to 1.07±0.04% day−1. Food conversion ratio (FCR) ranged from 1.09±0.02 to 0.92±0.03. The fastest growth and greatest final body weight were observed in the SAT treatment; however, the highest FCR, visceral fat index (VFI %) and hepatosomatic index (HSI %) were also observed in this treatment. Significant differences were found in specific growth rate and final mean weight between fish in the B/D and SAT treatments. B/N and B/DN feeding regimes appeared to result in the most favourable fish performance.

Fatty acid metabolism in the freshwater fish Murray cod (Maccullochella peelii peelii) deduced by the whole-body fatty acid balance method

The whole-body fatty acid balance method was used to investigate the fatty acid metabolism in Murray cod (Maccullochella peelii peelii) fed diets containing canola (CO) or linseed oil (LO). Murray cod were able to elongate and desaturate both 18 : 2n − 6 and 18 : 3n − 3. In fish fed the CO diet, 54.4% of the 18 : 2n − 6 consumed was accumulated, 38.5% oxidized and 6.4% elongated and desaturated to higher homologs. Fish fed the LO diet accumulated 52.9%, oxidized 37% and elongated and desaturated 8.6% of the consumed 18 : 3n − 3. The overall roles of n − 6 fatty acids appeared more important in Murray cod compared to other freshwater species. Murray cod also showed a preferential order of utilization of C18 fatty acid for energy production (18 : 3n − 3 > 18 : 2n − 6 > 18 : 1n − 9). Moreover, it is demonstrated that an increase in dietary 18 : 3n − 3 is directly responsible of increased desaturase activity and augmented saturated fatty acid accumulation in the fish body. The present study also suggests that, in the context of the possible maximization of the natural ability of fish to produce long chain polyunsaturated fatty acids, the whole-body approach can be considered well suited and informative and Murray cod is a suited candidate to fish oil replacement for its diets.

Effects of dietary oil source on growth and fillet fatty Acid composition of Murray cod, Maccullochella peelii peelii

The Murray cod, an Australian native freshwater fish, supports a relatively small but increasing aquaculture industry in Australia. Presently, there are no dedicated commercial diets available for Murray cod; instead, nutritionally sub-standard feeds formulated for other species are commonly used. The aim of the present investigation was to assess the suitability of two plant based lipid sources, canola oil (CO) and linseed oil (LO), as alternatives to fish oil for juvenile Murray cod. Five iso-nitrogenous, iso-calorific, iso-lipidic semi-purified experimental diets were formulated with 17% lipid originating from 100% cod liver oil (FO), 100% canola oil, 100% linseed oil and 1 : 1 blends of canola and cod liver oil (CFO) and 1 : 1 blends of linseed and cod liver oil (LFO). Each of the diets was fed to apparent satiation twice daily to triplicate groups of 50 Murray cod with initial mean weights of 6.45 ± 1.59 g for 84 days at 22 °C. Final mean weight, specific growth rate and daily feed consumption were significantly higher for the FO and LFO treatments compared to the LO treatment. Feed conversion and protein efficiency ratios were not significantly different amongst treatments. Experimental diets containing vegetable oil and vegetable oil blend(s) had significantly higher concentrations of n-6 fatty acids, predominantly in the form of linoleic acid (LA), while n-3 fatty acids were present in significantly higher concentrations in LO and LFO treatments. The fatty acid composition of Murray cod fillet was reflective of the dietary lipid source. Fillet of fish fed the FO was highest in EPA (20:5n-3), ArA (20:4n-6) and DHA (22:6n-3). Fish fed the CO diet had high concentrations of oleic acid (OlA) (192.2 ± 10.5 mg g lipid− 1), while the fillet of Murray cod fed the LO diet was high in α-linolenic acid (LnA) (107.1 ± 6.7 mg g lipid− 1). The present study suggests that fish oil can be replaced by up to 100% with canola oil and by up to 50% with linseed oil in Murray cod diets with no significant effect on growth.

Modification of tissue fatty acid composition in Murray cod (Maccullochella peelii peelii, Mitchell) resulting from a shift from vegetable oil diets to a fish oil diet.

The dynamics of fatty acid composition modifications were examined in tissues of Murray cod fed diets containing fish oil (FO), canola oil (CO) and linseed oil (LO) for a 25-week period and subsequently transferred to a FO (finishing/wash-out) diet for a further 16 weeks. At the commencement of the wash-out period, following 25 weeks of vegetable oil substitution diets, the fatty acid compositions of Murray cod fillets were reflective of the respective diets. After transfer to the FO diet, differences decreased in quantity and in numerousness, resulting in a revert to the FO fatty acid composition. Changes in percentages of the fatty acids and total accumulation in the fillet could be described by exponential equations and demonstrated that major modifications occurred in the first days of the finishing period. A dilution model was tested to predict fatty acid composition. In spite of a general reliability of the model (Y=0.9234X+0.4260, R²=0.957, P<0.001, where X is the predicted percentage of fatty acid; Y the observed percentage of fatty acid), in some instances the regression comparing observed and predicted values was markedly different from the line of equity, indicating that the rate of change was higher than predicted (i.e. Y=0.4205X+1.191, R²=0.974, P<0.001, where X is the predicted percentage of α-linolenic acid; Y the observed percentage of α-linolenic acid). Ultimately, using the coefficient of distance (D), it was shown that the fatty acid composition of fish previously fed the vegetable oil diets returned to the average variability of the fillet fatty acid composition of Murray cod after 70 or 97 days (LO and CO respectively).