Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

Three thousand litres of potable water is used each day to cook the Blue Swimmer Crabs which are caught by Abacus Fisheries. The cook water has a good flavour profile as it contains volatile compounds, proteins and minerals. Therefore the company wanted to know if it was feasible to utilise the crab cook water to produce a crab stock concentrate. The cadmium levels in the cook water were minimal and well below the maximum residue limit even when concentrated, therefore it did not pose a risk to consumers.

Proximate analysis conducted on the crab cook water determined that an increase in the crab cook load led to an increase in the components of total solids, protein, sodium and pH. The volume of crab cook water remains the same each day but the cook load does not. So, an equation was developed using the protein vs total solids regression line to determine the amount of water that had to be removed from the crab cook water to produce the stock at the desired concentration. Three different processing technologies (boiling, vacuum drying, freeze drying) used to produce stocks were evaluated and sensory analysis conducted on the products.

High quality fruits and vegetables grown in Carnarvon are being underutilised so they were used to produce two value added stocks using the crab stock base (herb flavoured crab stock and a chilli tomato crab stock). Sensory analysis conducted on the value added stocks indicated that both of the products were rated acceptable amongst the consumer panellists; however the chilli tomato crab stock was the most preferred.

In conclusion, the results from the research indicate that it is feasible to use the crab cook water to produce a crab stock concentrate and could also be potentially used to develop other food products.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.

The cDNA encoding for G-protein coupled receptor 54 (GPR54) was cloned from the brains of Southern Bluefin Tuna (SBT) and Yellowtail Kingfish (YTK). The SBT GPR54 has an open reading frame of 1134bp encoding a predicted 378 amino acid peptide, containing seven putative transmembrane domains, a 138 bp 5'UTR and 238 3'UTR. The partial YTK GPR54 cDNA contains 729 bp nucleotide sequence encoding 243 amino acid residues.

A RT-qPCR assay was developed for both SBT and YTK GPR54, as confirmed by primer specificity and high R2 values. The reference gene used, Hprt1, displayed consistent Cq values across most of the different sampling points, however some interaction between the reference gene expression and developmental stage was observed. GPR54 expression levels were determined for two cohorts, an immature group and a pubertal group, at two sampling points. RT-qPCR results showed that YTK brain GPR54 expression levels did not show sexual dimorphism and were significantly higher in the reproductively mature fish at a point past their peak spawning time compared with expression in the immature fish group.

The YTK gonad GPR54 expression level was significantly lower in males from the pubertal fish group during peak spawning time. In SBT, GPR54 expression profile in the brain and the gonad did not vary between immature and reproductively advanced fish. Analysis was carried out using both relative gene expression data and raw Cq. Alternative reference genes would need to be examined for the assay to be reliable across tissues and developmental time points.