Sexing accuracy and indicators of maturation status in captive Murray cod Maccullochella peelii peelii using non-invasive ultrasonic imagery

Macroscopic- and histological-based assessments of gonad condition were compared with ultrasound images to determine the feasibility of this technology as a non-invasive diagnostic tool for identifying sex and assessing maturation status of Murray cod. Four age-classes (1⁺, 2⁺, 3⁺ and 6⁺ years), were sub-sampled at monthly intervals throughout their annual reproductive cycle and scanned with a 5 MHz linear transducer. An interpretation of sex was made from the resulting images and maximum cross-sectional gonad diameter and area were recorded. Fish were subsequently dissected to confirm gender, and the weights and maturation status of gonads determined and then compared with their respective image profile. Ovaries of females were usually a distinctive feature in ultrasound images, being particularly obvious in older and/or more developed fish. In contrast, the identification of male testis was more problematic. Nonetheless, identifying sex from ultrasound images was consistently achieved by recording the presence/absence of a female ovary (96% total sexing accuracy). Maximum cross-sectional ovary diameter and area were highly correlated with gonad weight (r² = 0.90 and 0.89, respectively) suggesting that indices of maturation status, comparable to the gonadosomatic index (GSI), can be obtained non-destructively from ultrasound scans of females. A less distinct relationship occurred between these dimensions and weight of testes (r² = 0.41). Significant increases (P < 0.05) in mean gonad index (GI, calculated from gonad diameter) occurred for most gonad development stages. However, differences in mean GI between maturation stages were confounded by phenotypic variability, indicating that GI may be limited to population level studies. Nevertheless, ultrasound images of ovaries at each development stage were visually distinctive and enabled qualitative evaluations of maturity, thereby complementing quantitative GI assessments. Repeated serial-monitoring of the same population using ultrasound appears to have great potential for tracking maturation-induced changes in broodfish.

Age-related changes in ovarian characteristices, plasma sex steroids and fertility during pubertal development in captive female Murray cod Maccullochella peelii peelii

Age-related changes in ovarian development characteristics and plasma sex steroids in female Murray cod were examined throughout their second, third and fourth years of life to better understand the physiological and endocrine processes associated with puberty in this species in captivity. Spawning performance of 2+ and 3+ year old females was also assessed to identify ontogenetic differences in egg fertility. Puberty was acquired in 38% of 1+ year old females and 100% of age 2+ females. By age 3+, all females had developed full (adult) reproductive function. Ovarian development in pubertal fish was characterised by a rapid transition between cortical alveoli and lipid droplet oogenic phases, coinciding with significantly lower plasma 17β-oestradiol in age 2+ females (p < 0.05). Mean mature oocyte diameter (2.44 mm), post-fertilisation viability (30.80%) and hatchability (0.99%) of eggs from age 2+ females were significantly reduced relative to age 3+ adults (2.81 mm, 84.89% and 23.58%, respectively). Ovaries of pubertal Murray cod exhibited both vitellogenic and ovulatory capacities, yet functional abnormalities during secondary oocyte growth are likely to have contributed to poor egg fertility and consequently, evaluations of age-at-first maturity based on the presence of advanced ovarian stages may overestimate the reproductive potential of younger broodstock populations.

Cytochemical characterisation of the leucocytes and thrombocytes from murray cod (Maccullochella peelii peelii, Mitchell)

Cytochemistry has proven effective in differentiating specific cell lineages and elucidating their functional properties. This study utilised a range of cytochemical techniques to further investigate the leucocyte populations from Murray cod, an iconic Australian teleost fish species. This analysis provided clear insight into the structure and function of the leucocytes from this fish, which were found to be broadly similar to those of other fish species. However, some important differences were identified in Murray cod, such as the presence of naphthol AS chloroacetate esterase activity in the heterophil population, positive staining for periodic acid Schiff's, alkaline phosphatase and Sudan black B in the lymphocyte population, and a prevalent population of myeloid precursor cells.

Effects of alternate phases of fish oil and vegetable oil-based diets in Murray cod

Fish oil (FO)- and canola oil (CO)-based diets were regularly alternated in a daily cycle (amCO: alternation of CO in the morning and FO in the afternoon, and pmCO: alternation of FO in the morning and CO in the afternoon) or in a series of weekly cycles (2W: alternation of 2 weeks on CO and 2 weeks on FO, 4W: alternation of 4 weeks on CO and 4 weeks on FO), over a 16-week period in juvenile Murray cod (Maccullochella peelii peelii). No significant differences were observed between any of the treatments in relation to the final weight. However, fish subjected to the 2W schedule were larger (P>0.05) than all other treatments (37.2 ± 0.30 vs. 34.3 ± 0.58 in the control treatment). Fish receiving the 2W treatment had a significantly lower total net disappearance of eicosapentaenoic acid 20:5n-3 (EPA) and docosahexaenoic acid 22:6n-3 (62.1% and 24.0% respectively) compared with the control treatment (fish continuously fed a blend of 50% FO and 50% CO). Likewise, Murray cod receiving the amCO daily schedule had a significantly lower total net disappearance of EPA in comparison with the CD and pmCO treatments. These data point towards the existence of cyclical mechanisms relative to fatty acid utilization/retention.

Biometric, nutritional and sensory characteristic modifications in farmed Murray cod (Maccullochella peelii peelii) during the purging process

Intensively farmed, market-size Murray cod (~ 600 g), were purged (transferred into a clean water system and starved) and sampled at three day intervals for a total of 18 days (D0, D3, D6, D9, D12, D15 and D18). Purged fish lost from 6% (D3) to 14% (D18) body weight, and the weight loss was highly correlated to the number of days of purging/starvation. Condition factor and Hepatosomatic Index decreased significantly (P < 0.05) only after 18 days of purging compared to the control (D0). Fillet lipid content (%) did not vary during the trial. Eicosapentaenoic acid (EPA: 20:5 n−3) decreased and docosapentaenoic acid (DPA: 22:5 n−3) increased (P < 0.05) during the trial, while docosahexaenoic acid (DHA: 22:6 n−3) did not show any significant variation. Purging contributed positively to the improvement of the volatile flavour compound composition, with a significant (P < 0.05) reduction in total volatile aldehydes and an increase in total volatile hydrocarbons. Since no major differences were found between samples during the last stages of the purging process (D12, D15 and D 18), it is possible to conclude that, under these experimental conditions, 12 days is the minimum duration to obtain an improvement in the volatile compound profile of intensively farmed Murray cod whilst keeping body weight loss to a minimum.

Apparent in vivo Δ-6 desaturase activity, efficiency, and affinity are affected by total dietary C18 PUFA in the freshwater fish Murray cod

Dietary fatty acids are known to modulate fatty acid metabolism in fish. However, the innate capability of fish to bioconvert short chain fatty acids to health promoting long chain fatty acids (LCPUFA) is insufficient to compensate for a reduced dietary intake. While many studies have focused on the dietary regulation of the fatty acid bioconversion pathways, there is little known regarding the effects of the dietary levels of C18 polyunsaturated fatty acids (PUFA) on fatty acid metabolism. Here, we show a greater degree of apparent enzyme activity (Δ-6 desaturase) in fish fed a diet with higher amounts of dietary C18 PUFA. In particular, fish receiving high amounts of dietary C18 PUFA had a greater amount of Δ-6 desaturase activity acting on 18:3n-3 than 18:2n-6. However, with the gradual reduction of dietary C18 PUFA there was a shift in substrate preference of Δ-6 desaturase from 18:3n-3 to 18:2n-6. This information will provide valuable insight for the implementation of low fish oil diets, which permit the maintenance of n-3 LCPUFA levels in farmed Murray cod.

Traceability and discrimination among differently farmed fish : a case study on Australian murray cod

The development of traceability methods to distinguish between farmed and wild-caught fish and seafood is becoming increasingly important. However, very little is known about how to distinguish fish originating from different farms. The present study addresses this issue by attempting to discriminate among intensively farmed freshwater Murray cod originating from different farms (indoor recirculating, outdoor floating cage, and flow through systems) in different geographical areas, using a combination of morphological, chemical, and isotopic analyses. The results show that stable isotopes are the most informative variables. In particular, δ13C and/or δ15N clearly linked fish to a specific commercial diet, while δ18O linked fish to a specific water source. Thus, the combination of these isotopes can distinguish among fish originating from different farms. On the contrary, fatty acid and tissue proximate compositions and morphological parameters, which are useful in distinguishing between farmed and wild fish, are less informative in discriminating among fish originating from different farms.

Short-term food deprivation does not improve the efficacy of a fish oil finishing strategy in Murray cod

Two groups of fish (Maccullochella peelii peelii) were fed for a 90-day conditioning period on a canola oil diet (CO) or a fish oil diet (FO). Canola oil diet fed fish were then shifted to the FO diet for a 90-day finishing period. A variable period of  starvation (0, 5, 10 and 15 days) was introduced to reduce the initial lipid level of CO fed fish at the beginning of the finishing period and therefore accelerate the rate of recovery of FO-like fatty acids. During starvation, fish did not show  significant reduction in total lipid content, either in the fillet or whole body. At the end of the conditioning period, fatty acid composition of the diet was mirrored in fish tissues. These differences came close to levelling out following re-feeding, with the exception of n - 6 polyunsaturated fatty acids (PUFA). However, no  effects of the starvation periods on the final fatty acid make-up of fish were recorded. The results of this trial show that Murray cod, when subjected to a starvation period of up to 15 days, does not lose an appreciable quantity of lipid and, therefore, the tested starvation approach to reduce the initial level of lipid has to be considered unsuccessful. 

Biology and management of Murray cod reproduction within controlled environments

The emerging Murray cod aquaculture industry is currently restricted by a lack of consistent and year-round production of fingerlings. Biological information and reproductive technologies developed during this research have facilitated the development of indoor, environmentally-controlled breeding practices for this species, thereby enabling Murray cod juveniles to be produced on-demand.

Haematology of cultured Murray cod

Murray cod are Australia's largest freshwater fish: endangered in the wild, but supporting a boutique industry. This project studied the blood cells of Murray cod and those affected by the disfiguring chronic erosive dermatopathy. This has provided insight into the factors affecting Murray cod health.

Chronic ulcerative dermatopathy syndrome in Murray cod

Chronic ulcerative dermatopathy is a disease affecting Murray cod in aquaculture facilities that utilise groundwater. This study investigated the effects of this condition on key physiological processes and developed a cost-effective method of pre-treating the groundwater source that prevents the condition arising in Murray cod.