Species ID Project

Up to 10 individuals of a fin fish, shark, and crustacean species are collected and morphologically identified by Southeast Fisheries Science Center. Water-soluble sarcoplasmic proteins are extracted from the tissue; proteins are analyzed by microfluidic electrophoresis (Agilent Bioanalyzer 2100) to generate species-specific protein patterns. The protein patterns are entered into a database to determine which are the most abundant for each species. The proteins that are seen in all 10 individuals are transferred to the excel pattern matching library database that uses NSIL designed formulas to calculate the range of each protein (+/- 1.5 to 3%) and compare an unknown protein to pattern to those in the pattern matching library.

The pattern matching library is designed to hold the highly abundant proteins extracted from tissue collected from 10 individuals of a species and compare an unknown pattern for identification.

Specimens are collected and identified by the Southeast Fisheries Science center.

Protein pattern analysis is is conducted by NMFS OSF National Seafood Inspection Laboratory.

Species name, Species-specific protein masses, Sample protein masses, total # proteins, # of protein matches, Sample total of # of proteins, Library sample protein quantity ration, and similarity score.

All the proteins in the library are from authenticated species and the most abundant with-in 10 individuals.

The database is only assessable by being in the facility and use of a CAC card to sign-in to any facility computer. The IT coordinator assigns which personnel in the facility can enter into each network folder.

The database is only accessible through the facility's network and protected from public access by a firewall.

2010 Microsoft Excel

The number of proteins in the pattern matching library are species specific.

Individuals are authenticated morphologically by a fisheries biologist, documented and photographed for outside review.

Protein size can be affected by many differing environmental factors. Due to this variation, a protein size range (+/- 1.5 to 3%) is used.

The number of proteins in the pattern matching library are species specific.

If the individual is misidentified (morphologically), the protein patterns can shift in the wrong direction or some high abundant protein can be excluded that should not.

Each species in the library is compared to the library and others of that species not used as one of the to individuals is compared to verify the accuracy of the library.

The ideal number of species-specific proteins is greater than five. The highest amount of proteins extracted is 16 and then decreased to only include those observed in all 10 individuals.

Protein size can be affected by many differing environmental factors. Due to this variation, a protein size range (+/- 1.5 to 3%) is used.

Protein size can be affected by many differing environmental factors. Due to this variation, a protein size range (+/- 1.5 to 3%) is used.

On other analyses have been performed for comparison.

The pattern matching library compares all proteins of an unknown to the species specific proteins with-in the range (+/- 1.5 to 3%).

The height threshold is initially set to 5FU and raised incrementally for each individual to remove insignificant proteins until the total number of proteins is less than 16

Only proteins that are seen in all 10 individuals, and above the height threshold of 5FU are inserted into the pattern matching library.

Always.

The highly abundant proteins are determined, entered into the pattern matching library that performs the +/- 1.5 to 3% calculation and transcribed into a second worksheet to remove rounding extensions. All parts are checked by a second laboratory personnel.

NA

Limited access and QA

A specimen is caught and recorded on the vessel as its location, date and condition. After the specimen is brought to NSIL, it is identified by a fisheries biologist and the data recorded on the vessel is recorded into the database. The specimen is process and the number of vials is recorded.