Methods and systems for measuring growth rate in plant or aquatic animal species

Abstract

Method and systems for measuring growth rate in plant or aquatic animal species such as embryonic or adult fish. The methods and systems utilize the measurement of NADH2 production by detecting a colorimetric and fluorescent shift when a redox indicator such as resazurin is added to a sample. The colorimetric/fluorescent shift is indicative of the reduction of the redox indicator by NADH2. The methods and systems of the present invention may be used to predict growth potential of a plant or animal, and measuring the growth rate of said plant or animal may be helpful for identifying and selecting individuals within a group that have greater growth potential. The methods and systems of the present invention may help eliminate the need for special equipment (e.g., for measuring oxygen consumption), decrease variability of measures, and minimize the effects of external factors (feeding/hormonal status).

1 . A method of brood stock separation from a pool of individuals, said method comprising:
 a. measuring whole body metabolic rate in a cell sample from each individual, wherein metabolic rate is measured using a colorimetric/fluorescent redox assay comprising introducing a redox indicator to the cell sample at time T 0 ; measuring a fluorescence value at time T 0 ; measuring a fluorescence value at time T 1  after time T 0 ; and determining the change in fluorescence between T 1  and T 0  to determine a metabolic rate; and   b. separating the individuals in the group that had a metabolic rate in the 10% highest metabolic rates of the group from the remaining individuals in the group.   
     
     
         2 . The method of  claim 1 , wherein the pool of individuals comprises embryonic or juvenile aquatic organisms, cells, or plants. 
     
     
         3 . The method of  claim 2 , wherein the aquatic organisms are fish embryos. 
     
     
         4 . The method of  claim 2 , wherein the aquatic organisms are adult fish. 
     
     
         5 . The method of  claim 1 , wherein individuals in the group that had a metabolic rate in the 5% highest metabolic rates of the group are separated from the remaining individuals in the group. 
     
     
         6 . The method of  claim 1 , wherein the redox indicator comprises AlamarBlue®, resazurin, a tetrazolium dye, or PrestoBlue®. 
     
     
         7 . The method of  claim 1 , wherein T 1  is equal to T 0  plus 8 hours, T 0  plus 16 hours, or T 0  plus 24 hours. 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1 , wherein measuring a fluorescence value comprises using a fluorescent plate reader. 
     
     
         11 . The method of  claim 1 , wherein measuring a fluorescence value comprises obtaining a digital photograph of the cell sample and quantitating red, green, and blue color intensity, wherein the intensities are correlated with fluorescence to determine a quantitative measurement of metabolic rate. 
     
     
         12 . A method for predicting growth of individual organisms within a group, said method comprising:
 a. measuring metabolic rate in a cell sample of the organism by performing a colorimetric/fluorescent redox assay comprising introducing a redox indicator to the cell sample at time T 0 , measuring a fluorescence value at time T 0 , and measuring a fluorescence value at time T 1 ;   b. determining a metabolic rate by calculating the change in fluorescence between T 1  and T 0 ; and   c. comparing the metabolic rates of each of the organisms within the group; wherein a first organism with a higher metabolic rate than a second organism will be bigger, heavier, or both bigger and heavier at a time beyond a time associated with material effect.   
     
     
         13 . The method of  claim 12 , wherein the organism is a plant or aquatic animal. 
     
     
         14 . The method of  claim 13 , wherein the aquatic animal is a fish. 
     
     
         15 . The method of  claim 13 , wherein the organism is an embryonic aquatic organism. 
     
     
         16 . The method of  claim 13 , wherein the organism is an adult aquatic organism. 
     
     
         17 . The method of  claim 12 , wherein the redox indicator comprises AlamarBlue®, resazurin, a tetrazolium dye, or PrestoBlue®. 
     
     
         18 . The method of  claim 12 , wherein T 1  is equal to T 0  plus any given duration of time. 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 12 , wherein measuring a fluorescence or absorbance value is assessed using a spectrophotometer with or without fluorescent capabilities. 
     
     
         22 . The method of  claim 12 , wherein measuring a fluorescence value comprises obtaining a digital photograph of the cell sample and quantitating red, green, and blue color intensity, wherein the intensities are correlated with fluorescence to determine a quantitative measurement of metabolic rate. 
     
     
         23 . The method of  claim 12 , wherein the time beyond the time associated with maternal effect is harvest time. 
     
     
         24 . The method of  claim 12 , wherein the time beyond the time associated with maternal effect is 3 months or 8 months. 
     
     
         25 .- 32 . (canceled)