Strength and limitations of the study

5.7 Strength and limitations of the study

Š‡ •–”‡‰–Š• ‘ˆ –Š‹• •–—†› ƒ”‡ –Š‡ …‘ŽŽ‡…–‹‘ ‘ˆ •ƒ’Ž‡• ‹ Ž‘…ƒ–‹‘• –Šƒ– ™‘—Ž† ‘– „‡

ƒ……‡••‹„Ž‡™‹–Š‘—–Ž‘…ƒŽ…‘–ƒ…–ǤŠ‹•…‘—‹…ƒ–‹‘ƒ†‡‹–‡ƒ•‹‡”–‘ƒ””ƒ‰‡ˆ‹‡Ž†™‘”

ƒ†–‘”‡Ž›‘–Š‡‘„–ƒ‹‡†•ƒ’Ž‡•Ǥƒ˜‹‰‘™Ž‡†‰‡‘ˆ–Š‡Ž‘…ƒ–‹‘•ƒŽ•‘…‘–”‹„—–‡†–‘

–Š‡•—……‡••‘ˆ–Š‹•™‘”Ǥ

Š‡ ƒƒŽ›•‹• ‘ˆ •ƒ’Ž‡• ‹ –Š‹• •–—†› ™‡”‡ ’‡”ˆ‘”‡† ƒ– Š‹‰Š …ƒ’ƒ…‹–› ƒ† …‘’‡–‡–

Žƒ„‘”ƒ–‘”‹‡•™‹–Š•‹Žˆ—ŽŽƒ„•…‹‡–‹•–Ǥƒ††‹–‹‘ǡ–Š‡Žƒ”‰‡—„‡”‘ˆ…‘’‘—†•ƒƒŽ›•‡†

ˆ‘”‰ƒ˜‡ƒ„‡––‡”„ƒ…‰”‘—†ˆ‘”ƒ–Š‘”‘—‰Šƒ••‡••‡–‘ˆŠ‡ƒŽ–Š”‹••ǤŠ‡‹…Ž—•‹‘‘ˆ–Š‡

™‹†‡•—‹–‡‘ˆ…‘–ƒ‹ƒ–•ƒŽ•‘…‘–”‹„—–‡†–‘–Š‡‘™Ž‡†‰‡‘ˆ…‘–ƒ‹ƒ–‡š’‘•—”‡‹

ƒœƒ‹ƒǤ

‘”‡‘˜‡”ǡ ’‡”ˆ‘”‹‰ …‘–ƒ‹ƒ– ƒƒŽ›•‹• ‡•’‡…‹ƒŽŽ› ˆ‘” • ƒ– –Š‡ •ƒ‡ ƒ……”‡†‹–‡†

Žƒ„‘”ƒ–‘”›ǡ”‡†—…‡†–Š‡—…‡”–ƒ‹–›‘ˆ–Š‡”‡•—Ž–•’”‡•‡–‡†‹–Š‹••–—†›Ǥ —”–Š‡”‘”‡ǡ–Š‡

‘”–ŠǦ‘—–Š …‘ŽŽƒ„‘”ƒ–‹‘ …‘–”‹„—–‡† —…Š ‹ –Š‡ …ƒ’ƒ…‹–› „—‹Ž†‹‰ ƒ† –”ƒ•ˆ‡” ƒ†

•Šƒ”‹‰‘ˆ‘™Ž‡†‰‡Ǥ

Š‡Ž‹‹–ƒ–‹‘•‘ˆ–Š‹••–—†›‹…Ž—†‡–Š‡’‘Ž‹–‹…ƒŽ†‹•–—”„ƒ…‡•™Š‹…Š†‡Žƒ›‡†•ƒ’Ž‹‰‹–Š‡

„‡‰‹‹‰‘ˆ–Š‡•–—†›ƒ†–Š‡ƒ„‹Ž‹–›–‘ƒ……‡•••‘‡•‹–‡•ƒ†”‡†—…‡†–Š‡’‘••‹„‹Ž‹–›–‘

•ƒ’Ž‡ƒŽƒ”‰‡—„‡”‘ˆˆ‹•ŠǤ‘™—„‡”‘ˆ•‹–‡•ƒ†—‡˜‡—„‡”‘ˆˆ‹•Š•’‡…‹‡•–Šƒ–

™‡”‡ƒƒŽ›•‡†ǡƒ†‡‹–…‘’Ž‡š–‘ƒ‡…‘’ƒ”‹•‘„‡–™‡‡•’‡…‹‡•ƒ†•‹–‡•Ǥ‘™•ƒ’Ž‡

—„‡”ƒ†–Š‡Ž‘™ƒ‘—–‘ˆ•ƒ’Ž‡ƒ–”‹š™‡”‡Ž‹‹–‹‰ƒƒŽ›•‡•‘ˆ‘–Š‡”…‘’‘—†•Ǥ

‘”‡‘˜‡”ǡŽ‹‹–ƒ–‹‘•ˆ”‘–Š‡–‹‹‰‘ˆ•ƒ’Ž‹‰•‘‡–‹‡•ƒ†‡‹–†‹ˆˆ‹…—Ž––‘‘„–ƒ‹–Š‡

•ƒ’Ž‡•ˆ”‘–Š‡…Š‘•‡•‹–‡•Ǥƒ††‹–‹‘ƒŽ’”‘„Ž‡™ƒ•–Šƒ–ˆ‹•Šˆƒ”‡”••‘‡–‹‡•Šƒ†

‘–Š‡” ƒ””ƒ‰‡‡–• ˆ‘” †‡Ž‹˜‡”‹‰ ‘ˆ –Š‡ ˆ‹•Š ƒ– –Š‡ –‹‡ ™Š‡ ‹– ™ƒ• ƒ‰”‡‡† ‘ ˆ‘” –Š‡

’”‘Œ‡…––‘…‘‡ƒ†Šƒ”˜‡•–Ǥ

—”–Š‡”‘”‡ǡ•‘‡‘–Š‡”…Š‡‹…ƒŽ•‘ˆ‹–‡”‡•–™‡”‡‘–‹…Ž—†‡†‹–Š‡ƒƒŽ›•‹•„‡…ƒ—•‡‘ˆ

ˆ‹ƒ…‹ƒŽ Ž‹‹–ƒ–‹‘•ǡ •—…Š ƒ• †‹‘š‹•ǡ „—– ƒŽ•‘ •ǡ ‘”‰ƒ‘’Š‘•’Šƒ–‡• ƒ† ’›”‡–Š”‘‹†•Ǥ

Š‡”‡ ‹• ƒ —…‡”–ƒ‹–› ƒ„‘—– –Š‡ †‹‡–• ‘ˆ –Š‡ ƒœƒ‹ƒ ’‘’—Žƒ–‹‘ †—‡ –‘ Žƒ… ‘ˆ †‹‡–

•—”˜‡›•ƒ†”‡’‘”–•‡•’‡…‹ƒŽŽ›‘’‡”…ƒ’‹–ƒ…‘•—’–‹‘‘ˆˆ‹•Šˆ‘”†‹ˆˆ‡”‡–’‘’—Žƒ–‹‘

‰”‘—’•ǤŠ‹• ”‡•—Ž–‡†–‘—…‡”–ƒ‹–›‘‡•–‹ƒ–‡††‹‡–ƒ”›‹–ƒ‡‘ˆ…‘–ƒ‹ƒ–•ƒ†‘

Š‡ƒŽ–Š”‹•ƒ••‡••‡–Ǥ–Š‡’”‡•‡–•–—†›™‡—•‡†–Š‡Š‹‰Š‡”’‡”…ƒ’‹–ƒˆ‹•Š…‘•—’–‹‘

‡•–‹ƒ–‡†„› ˆ‘”ƒ•–ˆ”‹…ƒ’‘’—Žƒ–‹‘•ǡ™Š‹…Š‹‰Š–‘–„‡”‡’”‡•‡–ƒ–‹˜‡ˆ‘”–Š‡

‰‡‡”ƒŽ’‘’—Žƒ–‹‘Ǥ

Š‡ƒ••‡••‡–‘ˆ”‹•‘ˆ•ƒ†Š‡ƒ˜›‡–ƒŽ•–‘ƒœƒ‹ƒ•‹–Š‹••–—†›†‹†‘–‹…Ž—†‡

–Š‡…‘–”‹„—–‹‘•ˆ”‘‘–Š‡”ˆ‘‘†–›’‡•ƒ†‡˜‹”‘‡–•—…Šƒ•‹†‘‘”†—•–ǤŠ‡”‡ˆ‘”‡ǡ–Š‡

Ž‡˜‡Ž ‘ˆ ‡š’‘•—”‡ –‘ • ƒ† Š‡ƒ˜› ‡–ƒŽ• ˆ‘” ƒ ƒœƒ‹ƒ ‹• Ž‹‡Ž› –‘ „‡ Š‹‰Š‡” –Šƒ

‡•–‹ƒ–‡†ǤŠ—•ǡ–Š‡’‘–‡–‹ƒŽŠ‡ƒŽ–Š”‹••–‘ƒœƒ‹ƒ•ƒ›„‡Š‹‰Š‡”–Šƒ”‡’‘”–‡†‹–Š‹•

•–—†›Ǥ

6 MAIN CONCLUSIONS AND RECOMMENDATIONS

Š‡ƒ‹ˆ‹†‹‰•ˆ”‘–Š‹••–—†›™‡”‡–Šƒ–Ž‡˜‡Ž•‘ˆ•ƒ†Š‡ƒ˜›‡–ƒŽ•‹ˆƒ”‡†ƒ†

™‹Ž†ˆ‹•Šˆ”‘ƒœƒ‹ƒ˜ƒ”‹‡†™‹†‡Ž›„‡–™‡‡Ž‘…ƒ–‹‘•‹†‹…ƒ–‹‰‡š’‘•—”‡ˆ”‘˜ƒ”‹‘—•

•‘—”…‡•ǡ•—…Šƒ•†‹•…Šƒ”‰‡•ˆ”‘ƒ‰”‹…—Ž–—”ƒŽƒ…–‹˜‹–‹‡•ƒ†•‡™ƒ‰‡ǡ‡‹••‹‘ˆ”‘ƒ‹”’‘”–•

ƒ†‹’”‘’‡”†‹•’‘•ƒŽ‘ˆ…‘•—‡”’”‘†—…–•ǤŠ‡Ž‡˜‡Ž•‘ˆ•ƒ†„™‡”‡Š‹‰Š‡”‹™‹Ž†

ˆ‹•Š …‘’ƒ”‡† –‘ ˆƒ”‡† ˆ‹•Š ‹ ƒœƒ‹ƒǡ •—‰‰‡•–‹‰ –Šƒ– ™‹Ž† ˆ‹•Š ‹• ‘”‡ ‡š’‘•‡† –‘

’‘ŽŽ—–‹‘ –Šƒ– ˆƒ”‡† ˆ‹•ŠǤ ‡…‡ ƒ“—ƒ…—Ž–—”‡ ƒ› „‡ ‘”‡ ‡’Šƒ•‹œ‡†Ǥ ‘™‡˜‡”ǡ

Ž‡˜‡Ž•™‡”‡Š‹‰Š‡”‹ˆƒ”‡†ˆ‹•Š–Šƒ‹™‹Ž†ˆ‹•Šǡ’‘••‹„Ž›”‡Žƒ–‡†–‘ƒŽ‘…ƒŽ•‘—”…‡Ǥp,p’Ǧ

™ƒ•–Š‡†‘‹ƒ–…‘–ƒ‹ƒ–‹–Š‡ƒœƒ‹ƒˆ‹•Šƒ†–Š‡”ƒ–‹‘‘ˆp,p’Ǧ–‘p,p’-•—‰‰‡•–‡†„‘–Š”‡…‡–ƒ†Š‹•–‘”‹…ƒŽ—•‡‘ˆǤƒ†ɀǦ™‡”‡ˆ‘—†‹ͳͲͲΨ‘ˆ–Š‡

•ƒ’Ž‡•‘ˆˆƒ”‡†ƒ†™‹Ž†ˆ‹•Šˆ”‘ƒŽŽ•‹–‡••Š‘™‹‰ƒ™‹†‡†‹•–”‹„—–‹‘‘ˆ–Š‡…‘’‘—†•

‹ –Š‡ ƒœƒ‹ƒ ‡˜‹”‘‡–Ǥ Š‡ •–—†› ”‡˜‡ƒŽ‡† „ Ž‡˜‡Ž• ƒ„‘˜‡ • ‹ ͳͲͲΨ ‘ˆ –Š‡

•–—†‹‡†ˆ‹•Šˆ”‘ƒœƒ‹ƒǤŠ‡™‹†‡†‹•–”‹„—–‹‘‘ˆ„‹–Š‡ƒœƒ‹ƒ‡˜‹”‘‡–ƒ›

„‡ …ƒ—•‡† †—‡ –‘ Š‹•–‘”‹…ƒŽ —•‡ ‘ˆ Ž‡ƒ†‡† ‰ƒ•‘Ž‹‡ ƒ† ‹–•’”‡•‡…‡ ‹ ˜ƒ”‹‘—• …‘•—‡”

’”‘†—…–••—…Šƒ•’ƒ‹–ƒ†„ƒ––‡”‹‡•Ǥƒ††‹–‹‘ǡ–Š‡†‹ƒ…‡ƒ…—””‡–ƒ›…‘–”‹„—–‡

–‘ Š‹‰Š „ Ž‡˜‡Ž• ‹ ™‹Ž† ˆ‹•ŠǤ Š‡ ƒ••‡••‡– ‘ˆ Š—ƒ Š‡ƒŽ–Š ”‹• ˆ”‘ –Š‡ •–—†› ‘ˆ

ƒœƒ‹ƒˆ‹•Šǡ•—‰‰‡•–‘ƒ†˜‡”•‡Š‡ƒŽ–Š‡ˆˆ‡…–ˆ”‘…‘•—‹‰ˆƒ”‡†ƒ†™‹Ž†‹Žˆ‹•Š

ƒ†—ŽŽ‡–•ˆ”‘ƒœƒ‹ƒǤ

 –Š‡ ‹’‘”–‡† ˆ‹•Šˆ”‘Š‹ƒǡ•‹‰‹ˆ‹…ƒ– ˜ƒ”‹ƒ–‹‘ ‹Ž‡˜‡Ž• ƒ†’ƒ––‡”• ‘ˆ•ƒ†

Š‡ƒ˜›‡–ƒŽ•™‡”‡ˆ‘—†„‡–™‡‡–Š‡‹†‹˜‹†—ƒŽˆ‹•Šǡ•—‰‰‡•–‹‰–Šƒ––‹Žƒ’‹ƒˆ”‘–Š‡•ƒ‡

’ƒ…ƒ‰‡™‡”‡ˆ”‘†‹ˆˆ‡”‡–Ž‘…ƒ–‹‘•‹Š‹ƒǤŠ‡”ƒ–‹‘‘ˆp,p’Ǧ–‘p,p’Ǧ•—‰‰‡•–‡†

„‘–Š”‡…‡–ƒ†Š‹•–‘”‹…ƒŽ—•‡‘ˆ‹Š‹ƒǤŠ‡•–—†›ˆ‘—†Ž‘™Ž‡˜‡Ž•‘ˆ‹”‡š‹–Š‡

ͻͶΨ‘ˆ–Š‡ˆ‹•Š•ƒ’Ž‡•ƒ†–Šƒ––Š‡’ƒ––‡”‘ˆ•‘•–Ž‹‡Ž›”‡ˆŽ‡…–‡†—•‡‘ˆ–‡…Š‹…ƒŽ

‹š–—”‡‘ˆ‹–Š‡’ƒ•–ǤŠ‡•–—†›ƒŽ•‘•—‰‰‡•–•–Šƒ–‹’‘”–‹‰…‘‘†‹–‹‡•ˆ”‘Š‹ƒ

ƒ›„‡ƒ’ƒ–Š™ƒ›‘ˆ‹–”‘†—…‹‰…‘’‘—†•Ž‹‡ǦʹͲͻƒ†–‘–Š‡ƒœƒ‹ƒ

‡˜‹”‘‡–Ǥ••‡••‡–‘ˆŠ—ƒŠ‡ƒŽ–Š”‹••—‰‰‡•–‡†’‘••‹„Ž‡ƒ†˜‡”•‡Š‡ƒŽ–Š‡ˆˆ‡…–†—‡

–‘ǦǦͳͳͺƒ†ǦͳͲͷƒ†•ˆ”‘…‘•—’–‹‘‘ˆ–‹Žƒ’‹ƒˆ”‘Š‹ƒǤŽ–Š‘—‰Š–Š‡Ž‡˜‡Ž•

‘ˆ • ™‡”‡ ”‡Žƒ–‹˜‡Ž› Ž‘™ ‹ –Š‡ –‹Žƒ’‹ƒǡ ‹– ‹• ƒ Š‹‰ŠŽ› –‘š‹… ‡–ƒŽ ƒ† •Š‘™‡† ƒ Š‹‰Š

…‘–”‹„—–‹‘–‘–Š‡Šƒœƒ”†‹†‡šƒ†–Š‡ͻͷ^–Š’‡”…‡–‹Ž‡…ƒ…‡””‹•Ž‡˜‡Ž•™‡”‡–™‘–‹‡•

Š‹‰Š‡”–Šƒ–Š‡—’’‡”‡†‘ˆ–Š‡…ƒ…‡””‹•Ž‡˜‡Ž”ƒ‰‡ǤŠ‡‹’‘”–ƒ–‹‘‘ˆ–‹Žƒ’‹ƒˆ”‘

Š‹ƒƒ›„‡ƒƒ††‹–‹‘ƒŽ’ƒ–Š™ƒ›‘ˆ‡š’‘•—”‡–‘•ˆ‘”Š—ƒ•‹ƒœƒ‹ƒǡ‹ƒ††‹–‹‘

–‘‡š’‘•—”‡ˆ”‘‰”‘—†™ƒ–‡”•‘—”…‡•ǤŠ‡†‡–‡…–‹‘‘ˆ™‹†‡˜ƒ”›‹‰Ž‡˜‡Ž•ƒ†’ƒ––‡”•

‹ ‹†‹˜‹†—ƒŽ ˆ‹•Š ˆ”‘ ƒ ’ƒ…ƒ‰‡ ‘ˆ ‹’‘”–‡† –‹Žƒ’‹ƒ ˆ”‘ Š‹ƒǡ …ƒŽŽ• ˆ‘” ‹•’‡…–‘”• –‘

‘……ƒ•‹‘ƒŽŽ›‹…Ž—†‡‘‹–‘”‹‰‘ˆ’‘ŽŽ—–ƒ–•‹‹†‹˜‹†—ƒŽˆ‹•Š”ƒ–Š‡”–Šƒ–’‘‘Ž‡†•ƒ’Ž‡•Ǥ

Š‡ ƒ••‡••‡– ‘ˆ ˆ‹•Š Š‡ƒŽ–Š ”‹• ‹ –Š‹• •–—†› ”‡˜‡ƒŽ‡† –Šƒ– • ƒ† ‰ ƒ› ’‘•‡

’‘••‹„Ž‡ƒ†˜‡”•‡Š‡ƒŽ–Š‡ˆˆ‡…–•‹ˆ‹•Šƒ†ƒ›’‘•‡‡…‘Ž‘‰‹…ƒŽ Š‡ƒŽ–Š”‹•ƒ†–Š”‡ƒ–‡•

„‹‘†‹˜‡”•‹–›Ǥ —”–Š‡”‘‹–‘”‹‰•–—†‹‡•ƒ”‡”‡…‘‡†‡†Ǥƒ††‹–‹‘ǡ–Š‡•–—†›…ƒŽŽ•ˆ‘”

”‡‰—Žƒ”‘‹–‘”‹‰‘ˆ•ƒ†Š‡ƒ˜›‡–ƒŽ•‹ˆƒ”‡†ƒ†™‹Ž†ˆ‹•Š‹ƒœƒ‹ƒˆ‘”ˆ—”–Š‡”

†‡˜‡Ž‘’‡–‘ˆƒ“—ƒ…—Ž–—”‡ƒ†–Š‡™‡Žˆƒ”‡‘ˆ–Š‡™‹Ž†ˆ‹•Š•–‘…‹–Š‡…‘ƒ•–ƒŽ™ƒ–‡”•Ǥ

Ž–Š‘—‰Š–Š‹••–—†›”‡˜‡ƒŽ‡†•‘‡’‘–‡–‹ƒŽ…‘…‡”•‘Ž‡˜‡Ž•‘ˆ•ƒ†Š‡ƒ˜›‡–ƒŽ•‹

ƒœƒ‹ƒƒ†Š‹‡•‡ˆ‹•Šǡ–Š‡„‡‡ˆ‹–•‘ˆˆ‹•Š…‘•—’–‹‘•Š‘—Ž†„‡’—–‹‹†Ǥ

7 FUTURE PERSPECTIVE AND REFELECTIONS

—–—”‡’‡”•’‡…–‹˜‡•ƒ†”‡ˆŽ‡…–‹‘•ƒ”‡†‡•…”‹„‡†„‡Ž‘™Ǥ

x Š‡Ž‘™”ƒ–‹‘‘ˆp,p’ǦȀp,p’Ǧ‹•‘‡‘ˆ–Š‡ˆ‹•Š•ƒ’Ž‡•ǡ‹†‹…ƒ–‡†”‡…‡–—•‡

‘ˆ‹ƒœƒ‹ƒ™Š‹…Šƒ›•—‰‰‡•–‹ŽŽ‡‰ƒŽ—•‡‹ƒ‰”‹…—Ž–—”‡„‡•‹†‡•…‘„ƒ–‹‰

ƒŽƒ”‹ƒǤ ƒ”‡”••Š‘—Ž†„‡‰‹˜‡ƒ‘’’‘”–—‹–›ˆ‘””‡–—”‹‰‘Ž†’‡•–‹…‹†‡•–‘…•Ǥ

‘”†‡”–‘‡Ž‹‹ƒ–‡–Š‡‹ŽŽ‡‰ƒŽ•‘—”…‡•ǡ‡š‹•–‹‰”‡‰—Žƒ–‹‘•ˆ‘”—•‡‘ˆ„›–Š‡

‰‘˜‡”‡–ƒ†–Š‡–‘…Š‘Ž‘˜‡–‹‘•Š‘—Ž†„‡ˆ‘ŽŽ‘™‡†—’‘”‡•–”‹…–Ž›ǤŠ‹•

‡‡†•’”‘’‡”’‘Ž‹…›‘’‘ŽŽ—–ƒ–ƒƒ‰‡‡–Ǥ

x ‘‹–‘”‹‰ ’”‘‰”ƒ• ‘ˆ ‡› • ƒ† Š‡ƒ˜› ‡–ƒŽ• •Š‘—Ž† „‡ …‘†—…–‡† ‹ –Š‡

ƒœƒ‹ƒ ‡˜‹”‘‡– ƒ† ‹ ˆ‘‘† ’”‘†—…–• –‘ –”ƒ…‡ •‘—”…‡• ƒ† –‘ ‹’Ž‡‡–

‡ˆˆ‡…–‹˜‡‡ƒ•—”‡•ˆ‘”–Š‡‹”ƒƒ‰‡‡–ƒ†ƒ••‡••Š‡ƒŽ–Š”‹••Ǥ

x ‘”–”ƒ…‡ƒ„‹Ž‹–›’—”’‘•‡ǡ–Š‡‘”‹‰‹‘ˆ‹’‘”–‡†ˆ‹•Š•Š‘—Ž†„‡‘™‹†‡–ƒ‹Žƒ†

–Š‡‹•’‡…–‹‘ˆ‘”Šƒ”ˆ—Ž…Š‡‹…ƒŽ…‘–ƒ‹ƒ–•‹‹’‘”–‡†ˆ‹•Š•Š‘—Ž†‹…Ž—†‡

ƒ••‡••‡–‘ˆ‹†‹˜‹†—ƒŽˆ‹•Š•ƒ’Ž‡”ƒ–Š‡”–Šƒ’‘‘Ž‡†•ƒ’Ž‡•Ǥ

x —–—”‡ ‹˜‡•–‹‰ƒ–‹‘• •Š‘—Ž† ‹…Ž—†‡ ƒƒŽ›•‡• ‘ˆ ‡‡”‰‹‰ …‘–ƒ‹ƒ–• •—…Š ƒ•

ǡ •—„•–‹–—–‡• ˆ‘” „”‘‹ƒ–‡† ˆŽƒ‡ ”‡–ƒ”†ƒ–• ȋ •Ȍ ƒ† ˆŽ—‘”‹ƒ–‡†

”‡’‡ŽŽ‡–•ȋ •Ȍƒ†’‡•–‹…‹†‡•ǡŽ‹‡’›”‡–Š”‘‹†•ǡ…ƒ”„ƒƒ–‡•ƒ†‘–Š‡”–‘š‹…ƒ–•

Ž‹‡’Žƒ•–‹…‹œ‡”•ȋ’Š‡‘Ž•ǡ’Š–ŠƒŽƒ–‡•Ȍ‘–‹…Ž—†‡†‹–Š‹••–—†›Ǥ

x †ƒ–ƒ„ƒ•‡•Š‘—Ž†„‡†‡˜‡Ž‘’‡†–‘‘‹–‘”Ž‡˜‡Ž•ƒ†–”‡†•‘ˆ•‹ƒœƒ‹ƒ–‘

—†‡”•–ƒ†–Š‡†›ƒ‹…•ƒ†‹’Ž‡‡–ƒ–‹‘•‘ˆ”‡•–”‹…–‹‘•ƒ†„ƒ•Ǥ

x ƒ–‹‘ƒŽ†‹‡–•—”˜‡›••Š‘—Ž†„‡’‡”ˆ‘”‡†–‘‹†‡–‹ˆ›Š‹‰ŠǦ”‹•’‘’—Žƒ–‹‘‰”‘—’•

–‘‰‡–Š‡” ™‹–Š ‘‹–‘”‹‰ ’”‘‰”ƒ• ‘ Ž‘‰Ǧ–‡” Š‡ƒŽ–Š ‡ˆˆ‡…–• ‘ˆ ’‘ŽŽ—–ƒ–• –‘

Š—ƒ•Ǥ

x …”‡ƒ•‡‘ˆ’—„Ž‹…ƒ™ƒ”‡‡••‘–Š‡’‘••‹„Ž‡Š‡ƒŽ–Š”‹••”‡Žƒ–‡†–‘‡š’‘•—”‡–‘•ǡ

Š‡ƒ˜› ‡–ƒŽ• ƒ† ‘–Š‡” –‘š‹… …‘’‘—†• ‹ ƒœƒ‹ƒ •Š‘—Ž† „‡ †‘‡ –Š”‘—‰Š

•…‹‡–‹ˆ‹…™‘”•Š‘’•ƒ†•›’‘•‹—•ǡ‡†‹ƒ…‘˜‡”ƒ‰‡ƒ†‡†—…ƒ–‹‘ƒ–•…Š‘‘Ž•Ǥ

x ‘”–ŠǦ‘—–Š…‘ŽŽƒ„‘”ƒ–‹‘•Š‘—Ž†„‡•–”‡‰–Š‡‡†‹‘”†‡”–‘‹’”‘˜‡–Š‡•…‹‡–‹ˆ‹…

‘™Ž‡†‰‡ ƒ† „—‹Ž†‹‰ …ƒ’ƒ…‹–› ‘ •–—†‹‡• ‘ ’‘ŽŽ—–ƒ–•ǡ –Š‡‹” †‹•–”‹„—–‹‘• ƒ†

ƒ••‘…‹ƒ–‡†Š‡ƒŽ–Š”‹••Ǥ

x —Ž–‹†‹•…‹’Ž‹ƒ”›”‡•‡ƒ”…Š•—…Šƒ•…‘ŽŽƒ„‘”ƒ–‹‘„‡–™‡‡’—„Ž‹…Š‡ƒŽ–Šǡƒ–Š”‘’‘Ž‘‰›

ƒ†ƒ–—”ƒŽ•…‹‡…‡•Š‘—Ž†„‡‡•–ƒ„Ž‹•Š‡†‹‘”†‡”–‘—†‡”•–ƒ†–Š‡•‘…‹ƒŽƒ•’‡…–•

‘ˆ’‘ŽŽ—–ƒ–•‡š’‘•—”‡–‘–Š‡’‘’—Žƒ–‹‘Ǥ

x —–—”‡•–—†‹‡••Š‘—Ž†–ƒ‡‹–‘…‘•‹†‡”ƒ–‹‘–Š‡‹’ƒ…–‘ˆ…Ž‹ƒ–‡…Šƒ‰‡‘–Š‡

‰Ž‘„ƒŽƒ†Ž‘…ƒŽ†‹•–”‹„—–‹‘‘ˆ•ƒ†Š‡ƒ˜›‡–ƒŽ•ƒ††‹”‡…–ˆ‘…—•‘–Š‹•ƒ”‡ƒǡ

„‡…ƒ—•‡–Š‡‹ˆŽ—‡…‡‘ˆ…Ž‹ƒ–‡…Šƒ‰‡‘–Š‡•‡’‘ŽŽ—–ƒ–•‹•‘–ˆ—ŽŽ›—†‡”•–‘‘†Ǥ x ‘”ˆ‹•Šǡˆ—”–Š‡”•–—†‹‡•‘–Š‡Ž‡˜‡Ž•ƒ†‡ˆˆ‡…–•‘ˆ’‘ŽŽ—–ƒ–•‹ˆ‹•Šƒ†–Š‡‹”–”‘’Š‹…

–”ƒ•ˆ‡”‹–Š‡ƒ”‹‡‡…‘•›•–‡•Š‘—Ž†„‡‡…‘—”ƒ‰‡†Ǥ

8 REFERENCES

ŽǦ•ƒ™ƒˆ‹ǡǤǤǤǡƒ‰ǡǤǡƒǡǤǡʹͲͳ͹Ǥƒ†‹—ƒ……——Žƒ–‹‘ƒ†‹–•Š‹•–‘Ž‘‰‹…ƒŽ‡ˆˆ‡…–

‘ „”ƒ‹ ƒ† •‡Ž‡–ƒŽ —•…Ž‡ ‘ˆ œ‡„”ƒˆ‹•ŠǤ Ǥ ‡ƒ˜› ‡–Ǥ ‘š‹…Ǥ ‹•Ǥ Ͳʹǡ ͳȂ͸Ǥ

†‘‹ǣͳͲǤʹͳ͹͸͹ȀʹͶ͹͵Ǧ͸Ͷͷ͹ǤͳͲͲͲͳ͹

•ƒ–‡ǡǤǤǡƒƒŠƒ•Š‹ǡǤǡ–ƒ‹ǡǤǡ•‘„‡ǡǤǡ‡˜ƒƒ–ŠƒǡǤǡ—–‘ǡǤǡ‰›ƒ™ƒŠǡǤǤǡ†—Ǧ

—‹ǡǤǡ—„”ƒƒ‹ƒǡǤǡƒƒ„‡ǡǤǡʹͲͳ͵Ǥ……—””‡…‡‘ˆŠƒŽ‘‰‡ƒ–‡†…‘–ƒ‹ƒ–•‹

‹Žƒ†ƒ†…‘ƒ•–ƒŽˆ‹•Šˆ”‘Šƒƒǣ‡˜‡Ž•ǡ†‹‡–ƒ”›‡š’‘•—”‡ƒ••‡••‡–ƒ†Š—ƒ

Š‡ƒŽ–Š ‹’Ž‹…ƒ–‹‘•Ǥ …‘–‘š‹…‘ŽǤ ˜‹”‘Ǥ ƒˆǤ ͻͶǡ ͳʹ͵Ȃͳ͵ͲǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡…‘‡˜ǤʹͲͳ͵ǤͲͷǤͲͲͺ

ǡʹͲͳͶǤ‰‡…›ˆ‘”–‘š‹…•—„•–ƒ…‡•ƒ††‹•‡ƒ•‡”‡‰‹•–”›Ǥ‘Ž›…ŠŽ‘”‹ƒ–‡†„‹’Š‡›Ž•

ȋ•Ȍ–‘š‹…‹–›ǣŠƒ–”‡’‘Ž›…ŠŽ‘”‹ƒ–‡†„‹’Š‡›Ž•ȋ•Ȍǫȏ‘…—‡–ȐǤ

Š––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ…•‡Ȁ…•‡Ǥƒ•’ǫ…•‡α͵ͲƬ’‘αͶȋƒ……‡••‡†ͳʹǤͳͲǤͳͺȌǤ ǡʹͲͳͳǤ‰‡…›ˆ‘”–‘š‹…•—„•–ƒ…‡•ƒ††‹•‡ƒ•‡”‡‰‹•–”›Ǥ—„•–ƒ…‡ǦŽ‹•–‹‰’ƒ‰‡

ȏ ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ•—„•–ƒ…‡•Ȁ‹†‡šǤƒ•’͓

ȋƒ……‡••‡†͵ǤͳͳǤͳͺȌǤ

ǡ ʹͲͲͺǤ ‰‡…› ˆ‘” –‘š‹… •—„•–ƒ…‡• ƒ† †‹•‡ƒ•‡ ”‡‰‹•–”›Ǥ ‘š‹…‘Ž‘‰‹…ƒŽ ’”‘ˆ‹Ž‡ ˆ‘”

ƒŽ—‹— ȋŽȌ ȏ ‘…—‡–ȐǤ

Š––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ‘š”‘ˆ‹Ž‡•Ȁ–’ʹʹǤ’†ˆȋƒ……‡••‡†͵ǤͳͳǤͳͺȌǤ

ǡʹͲͲ͹ƒǤ‰‡…›ˆ‘”–‘š‹…•—„•–ƒ…‡•ƒ††‹•‡ƒ•‡”‡‰‹•–”›Ǥ‘š‘…‘Ž‘‰‹…ƒŽ’”‘ˆ‹Ž‡‘ˆŽ‡ƒ†

ȋ„Ȍ ȏ ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ–‘š’”‘ˆ‹Ž‡•Ȁ–’ͳ͵Ǥ’†ˆ

ȋƒ……‡••‡†͵ǤͳͳǤͳͺȌǤ

ǡʹͲͲ͹„Ǥ‰‡…›ˆ‘”–‘š‹…•—„•–ƒ…‡•ƒ††‹•‡ƒ•‡”‡‰‹•–”›Ǥ‘š‹…‘Ž‘‰‹…ƒŽ’”‘ˆ‹Ž‡ˆ‘”

ƒ”•‡‹… ȋ•Ȍ ȏ ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ–‘š’”‘ˆ‹Ž‡•Ȁ–’ʹǤ’†ˆ

ȋƒ……‡••‡†͵ǤͳͳǤͳͺȌǤ

ǡʹͲͲͷǤ‰‡…›ˆ‘”–‘š‹…•—„•–ƒ…‡•ƒ††‹•‡ƒ•‡”‡‰‹•–”›Ǥ—„Ž‹…Š‡ƒŽ–Š•–ƒ–‡‡–ˆ‘”

‹…‡Žȋ‹Ȍȏ‘…—‡–ȐǤŠ––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ‘š”‘ˆ‹Ž‡•Ȁ–’ͳͷǦ…ͳǦ

„Ǥ’†ˆȋƒ……‡••‡†͵ǤͳͳǤͳͺȌǤ

ǡ ͳͻͻͻǤ ‰‡…› ˆ‘” –‘š‹… •—„•–ƒ…‡• ƒ† †‹•‡ƒ•‡ ”‡‰‹•–”›Ǥ ‘š‹…‘Ž‘‰‹…ƒŽ ’”‘ˆ‹Ž‡ ˆ‘”

‡”…—”›ȋ‰Ȍȏ‘…—‡–ȐǤŠ––’•ǣȀȀ™™™Ǥƒ–•†”Ǥ…†…Ǥ‰‘˜Ȁ‘š”‘ˆ‹Ž‡•Ȁ–’Ͷ͸Ǧ …ͳǦ„Ǥ’†ˆȋƒ……‡••‡†ͳͳǤ͵ǤͳͺȌǤ

—–ŠƒǡǤǤǡʹͲͳͷǤ•‡‘ˆˆ‹•Šƒ•„‹‘Ǧ‹†‹…ƒ–‘”‘ˆ–Š‡‡ˆˆ‡…–•‘ˆŠ‡ƒ˜›‡–ƒŽ•’‘ŽŽ—–‹‘ǤǤ

“—ƒ…Ǥ‡•Ǥ‡˜ǤͲ͸Ǥ†‘‹ǣͳͲǤͶͳ͹ʹȀʹͳͷͷǦͻͷͶ͸ǤͳͲͲͲ͵ʹͺ

ƒ‰ƒ”‹ƒ‘ǡ ǤǤǡ ͳͻͻͳǤ ‹‘Ž‘‰› ‘ˆ ‹Žˆ‹•Š ȋChanos chanos ‘”••ƒŽȌǤ “—ƒ…—Ž–—”‡

‡’ƒ”–‡–ǡ‘—–Š‡ƒ•–•‹ƒ ‹•Š‡”‹‡•‡˜‡Ž‘’‡–‡–‡”ǡŽ‘‹Ž‘ǡŠ‹Ž‹’’‹‡•Ǥ

ƒœ‹ǡ ǤǤǡ•ƒ‹ǡǤǤǡ‘Šƒ‡†ǡǤǤǡʹͲͳͷǤ‹•–”‹„—–‹‘‘ˆŠ‡ƒ˜›‡–ƒŽ•‹•‘‹Ž•‹–Š‡

˜‹…‹‹–›‘ˆ–Š‡’”‘’‘•‡†—Œ——”ƒ‹—‹‡‹ƒœƒ‹ƒǤ˜‹”‘Ǥ‘ŽŽ—–ǤͶǡͶʹȂͷͲǤ

†‘‹ǣͳͲǤͷͷ͵ͻȀ‡’Ǥ˜Ͷ͵’Ͷʹ

‡”‰ǡ Ǥǡ ”ƒ—‰‡”—†ǡ Ǥǡ ‘—”‹œƒ†‡ŠǦ‹ŽŽƒ„ƒ†‹ǡ Ǥǡ Ž•˜‹ǡ ǤǤǡ ǫ”‡ǡ ǤǤǡ Ž‡•–”ǫǡ Ǥǡ

‘’•–ƒ†ǡǤǡ‹‡”ǡǤǤǡ›…Š‡ǡǤǤǡʹͲͳ͸Ǥ†‘…”‹‡ ‡ˆˆ‡…–•‘ˆ ”‡ƒŽǦŽ‹ˆ‡ ‹š–—”‡• ‘ˆ

’‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–• ȋȌ‹‡š’‡”‹‡–ƒŽ ‘†‡Ž• ƒ†™‹Ž†ˆ‹•ŠǤǤ‘š‹…‘ŽǤ

˜‹”‘Ǥ‡ƒŽǤǦƒ”–—””Ǥ••—‡•͹ͻǤ†‘‹ǣͳͲǤͳͲͺͲȀͳͷʹͺ͹͵ͻͶǤʹͲͳ͸Ǥͳͳ͹ͳͻͺͲ

‡”‰ǡǤƒ†‡ǡ‹”„ƒ—ǡǤǤǡ‡‹•‘ǡǤǡ‡‹–‘ǡǤǡ ƒ”Žƒ†ǡǤǡ ‡‡Ž‡›ǡǤǡ ‹‡†Ž‡”ǡǤǡ

ƒƒ••‘ǡ Ǥǡ ƒ„‡”‰ǡ Ǥǡ ƒ™•ǡ Ǥǡ ‘•‡ǡ Ǥǡ ƒˆ‡ǡ Ǥǡ …Š”‡ǡ Ǥǡ ‘Š›ƒƒǡ Ǥǡ

”‹–•…Š‡”ǡǤǡ—‘‹•–‘ǡǤǡ›•Ž‹†ǡǤǡƒŽ‡”ǡǤǡ‡–‡”•‘ǡǤǤǡʹͲͲ͸ǤŠ‡ʹͲͲͷ‘”Ž†

‡ƒŽ–Š”‰ƒ‹œƒ–‹‘”‡‡˜ƒŽ—ƒ–‹‘‘ˆŠ—ƒƒ†ƒƒŽ‹ƒ–‘š‹…‡“—‹˜ƒŽ‡…›ˆƒ…–‘”•

ˆ‘” †‹‘š‹• ƒ† †‹‘š‹ǦŽ‹‡ …‘’‘—†•Ǥ ‘š‹…‘ŽǤ …‹Ǥ ͻ͵ǡ ʹʹ͵ȂʹͶͳǤ

†‘‹ǣͳͲǤͳͲͻ͵Ȁ–‘š•…‹ȀˆŽͲͷͷ

‹–ƒŽƒǡǤ Ǥǡ™‡›—‰ƒǡǤǡƒ‘‘ǡǤǤǡʹͲͲͻǤ‡˜‡Ž•‘ˆŠ‡ƒ˜›‡–ƒŽ•ƒ†…›ƒ‹†‡‹•‘‹ǡ

•‡†‹‡– ƒ† ™ƒ–‡” ˆ”‘–Š‡ ˜‹…‹‹–› ‘ˆ ‘”–Š ƒ”ƒǡ ‘Ž† ‹‡ ‹ ƒ”‹‡ †‹•–”‹…–ǡ

ƒœƒ‹ƒǤ

‘”ƒǡǤǤǡ‡…ǦƒŠǡǤǤǡ†‡ ƒ‰‡”ǡǤǡƒ‰‡ƒƒ”ǡǤǤǡ‘—™ƒǡǤǡƒ”Š‘‘”ǡ ǤǤǤǡƒ–”‹…ǡǤǤǡƒ†‡„‡”‰ǡǤǤǡ‘”–‡ƒ’ǡǤǡŽ—„‡”‰ǡǤǡ‹‹•ǡǤǡ‡‰‘—ǡ Ǥǡ—‰‡”ǡǤǡ‹ƒ‰‹ǡǤǡ‡‹†‡ŽǡǤǤǡʹͲͳ͹Ǥ†‘…”‹‡†‹•”—’–‘”•ƒ†Š‡ƒŽ–Š‡ˆˆ‡…–•‹

ˆ”‹…ƒǣ …ƒŽŽ ˆ‘” ƒ…–‹‘Ǥ ˜‹”‘Ǥ ‡ƒŽ–Š ‡”•’‡…–Ǥ ͳʹͷǡ ͲͺͷͲͲͷǦͳǦͲͺͷͲͲͷǦͳͲǤ

†‘‹ǣͳͲǤͳʹͺͻȀͳ͹͹Ͷ

‘•…ŠǡǤǤǡǯ‡‹ŽŽǡǤǡ‹‰‰‡ǡǤǤǡ‡”™ƒ–ŠǡǤǤǡ‘ˆˆƒǡǤǤǡʹͲͳ͸Ǥ‡ƒ˜›‡–ƒŽ•‹ƒ”‹‡

ˆ‹•Š ‡ƒ– ƒ† …‘•—‡” Š‡ƒŽ–Šǣ ”‡˜‹‡™Ǥ Ǥ …‹Ǥ ‘‘† ‰”‹…Ǥ ͻ͸ǡ ͵ʹȂͶͺǤ

†‘‹ǣͳͲǤͳͲͲʹȀŒ•ˆƒǤ͹͵͸Ͳ

”‡‹˜‹ǡǤǡ”‹–ƒ‰‡ǡǤǤǡƒ‹ƒǡ Ǥǡ™‡‡–ƒǡǤǤǡ‘‡•ǡǤǤǡʹͲͳ͸Ǥ”ƒ…‹‰–Š‡‰Ž‘„ƒŽ

†‹•–”‹„—–‹‘ ‘ˆ ’‡”•‹•–‡– ‘”‰ƒ‹… ’‘ŽŽ—–ƒ–• ƒ……‘—–‹‰ ˆ‘” ‡Ǧ™ƒ•–‡ ‡š’‘”–• –‘

†‡˜‡Ž‘’‹‰”‡‰‹‘•Ǥ˜‹”‘Ǥ…‹Ǥ‡…Š‘ŽǤͷͲǡ͹ͻͺȂͺͲͷǤ†‘‹ǣͳͲǤͳͲʹͳȀƒ…•Ǥ‡•–Ǥͷ„ͲͶʹʹ͸

”‡˜‹ǡǤǤǡͳͻ͹ͺǤƒ•…Š”‘ƒ–‘‰”ƒ’Š‹…‡–Š‘†ˆ‘”–Š‡†‡–‡”‹ƒ–‹‘‘ˆ‘”‰ƒ‘…ŠŽ‘”‹‡

’‡•–‹…‹†‡•‹Š—ƒ‹ŽǤ—ŽŽǤ˜‹”‘Ǥ‘–ƒǤ‘š‹…‘ŽǤʹͲǡʹͺͳȂʹͺ͸Ǥ

—‰ƒŽƒǡǤǤǡƒ…Š‹™ƒǡǤ ǤǡŠ‹ŽŽƒǡǤǤǡʹͲͳ͹Ǥ‘…‡–”ƒ–‹‘ƒ†‹‘ƒ‰‹ˆ‹…ƒ–‹‘‘ˆ‡ƒ˜›

‡–ƒŽ•‹‹‘–ƒ‘ˆ–Š‡‘ƒ•–ƒŽƒ”‹‡”‡ƒ•‘ˆƒœƒ‹ƒǤǤ˜‹”‘Ǥ…‹Ǥ‰Ǥ͸ǡͶͲ͸Ȃ ͶʹͶǤ†‘‹ǣͳͲǤͳ͹ʹ͸ͷȀʹͳ͸ʹǦͷʹ͸͵ȀʹͲͳ͹ǤͲͺǤͲͲʹ

—”‹ƒǡǤǡƒƒ”ƒ–•ƒ‹ƒǡǤǡƒƒŽŽ‹ǡǤǡ‹ƒ‰ǡǤǤǡ‘†‡•ǡǤǡ”ƒ„‹…ǡǤǡ‡Ž‹•‡ǡǤǡ—”‡ǡǤǡ

‘Žƒ”‘˜ƒǡǤǡŽƒ„‡ǡǤǡƒ†ƒǡǤǡʹͲͳͺǤ‘’Ž‡š‡ˆˆ‡…–•‘ˆ’‘ŽŽ—–‹‘‘ˆ‹•Š‹ƒŒ‘”

”‹˜‡”• ‹ –Š‡ œ‡…Š ‡’—„Ž‹…Ǥ …‘–‘š‹…‘ŽǤ ˜‹”‘Ǥ ƒˆǤ ͳ͸Ͷǡ ͻʹȂͻͻǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡…‘‡˜ǤʹͲͳͺǤͲ͹ǤͳͲͻ

ǡʹͲͲ͹Ǥ’ƒ…–‘ˆƒ”‹…—Ž–—”‡‘…‘ƒ•–ƒŽ‡…‘•›•–‡•Ǥ‘”•Š‘’‘‘‰”ƒ’Š•

‘͵ʹǤ

Žƒ”•‘ǡǤǤǡ‘”†„‡”‰ǡǤ Ǥǡƒ‰‡”ǡǤǤǡͳͻͺͷǤ‡’”‘†—…–‹˜‡ƒ††‡˜‡Ž‘’‡–ƒŽ–‘š‹…‹–›‘ˆ

‡–ƒŽ•Ǥ …ƒ†Ǥ Ǥ ‘”Ǥ ˜‹”‘Ǥ ‡ƒŽǤ ͳͳǡ ͳͶͷȂͳͷͶǤ †‘‹ǣͳͲǤͳͲͳ͸Ȁͻ͹ͺǦͲͳʹ͵͸ͻͶͳ͵Ǧ

͵ȀͷͲͲ͸͹Ǧ͹

‘‹„”ƒǡǤǤǡ‡‹•Ǧ‡”‹“—‡•ǡǤǤǡʹͲͲ͹Ǥ‹Žƒ’‹ƒŽƒ”˜ƒ‡ƒ”‘…Ž‘”ͳʹͷͶ‡š’‘•—”‡ǣˆˆ‡…–•‘

‰‘ƒ†• ƒ† …‹”…—Žƒ–‹‰ –Š›”‘‹† Š‘”‘‡• †—”‹‰ ƒ†—Ž–Š‘‘†Ǥ —ŽŽǤ ˜‹”‘Ǥ ‘–ƒǤ

‘š‹…‘ŽǤ͹ͻǡͶͺͺȂͶͻ͵Ǥ†‘‹ǣͳͲǤͳͲͲ͹Ȁ•ͲͲͳʹͺǦͲͲ͹ǦͻʹͺͺǦʹ

‘””‡‹ƒǡǤǤǡƒ”…‹œ‘ǡ ǤǤǡ‹ƒ…Š‹‹ǡǤǡ‘”‡‹”ƒǡǤǤǡʹͲͳͲǤŽ—‹—ƒ• ƒ‡†‘…”‹‡

†‹•”—’–‘”‹ˆ‡ƒŽ‡‹Ž‡–‹Žƒ’‹ƒȋOreochromis niloticusȌǤ‘’Ǥ‹‘…Š‡ǤŠ›•‹‘ŽǤǡƒ”–

ͳͷͳǡͶ͸ͳȂͶ͸͸Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…„’…ǤʹͲͳͲǤͲʹǤͲͲʹ

”‹‹‘ǡǤǤǡʹͲͲͻǤŠŽ‘”‹ƒ–‡†’‡•–‹…‹†‡•ǣŠ”‡ƒ–•–‘Š‡ƒŽ–Šƒ†‹’‘”–ƒ…‡‘ˆ†‡–‡…–‹‘Ǥ

Ž–‡”Ǥ ‡†Ǥ ‡˜Ǥ ͳͶǡ ͵Ͷ͹Ȃ͵ͷͻǤ †‘‹ǣͲͲͲʹǦͻ͸ͳͲȋͲ͹ȌͲͲʹ͵ͻǦͷ ȏ’‹‹Ȑ̳”ͳͲǤͳͲͳ͸ȀŒǤƒŒ•—”‰ǤʹͲͲ͸ǤͳͲǤͲʹ͸

ƒ”‡”—†ǡǤǤǡ–—ƒǡǤǡ—‡ǡǤǡŒ‡”•‡Ž‹—•ǡǤǡŽ›ǡǤǡ”ƒ™‡ǡǤǤǡ‡…‡”ǡǤǡʹͲͲ͸Ǥ

‹‡–ƒ”› ‹–ƒ‡ ‡•–‹ƒ–‹‘• ‘ˆ ‘”‰ƒ‘ŠƒŽ‘‰‡ …‘–ƒ‹ƒ–• ȋ†‹‘š‹•ǡ ǡ ƒ†

…ŠŽ‘”‹ƒ–‡† ’‡•–‹…‹†‡•ǡ‡Ǥ‰Ǥ Ȍ „ƒ•‡† ‘ ™‡†‹•Š ƒ”‡– „ƒ•‡– †ƒ–ƒǤ ‘‘† Š‡Ǥ

‘š‹…‘ŽǤͶͶǡͳͷͻ͹Ȃͳ͸Ͳ͸Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤˆ…–ǤʹͲͲ͸ǤͲ͵ǤͲͳͳ

‘•ƒ–‘•ǡǤǤǡƒ˜ƒŽ…ƒ–‡ǡǤǤǤǡƒ—•‡”Ǧƒ˜‹•ǡǤǤǡ‘’‡•ǡǤǤǡƒ‘•–ƒƒ––‘•ǡǤǤǤǤǡ ʹͲͳ͸Ǥˆˆ‡…–•‘ˆ•—„ǦŽ‡–ŠƒŽƒ†…Š”‘‹…Ž‡ƒ†…‘…‡–”ƒ–‹‘•‘„Ž‘‘†ƒ†Ž‹˜‡”Ǧ

ƒ…–‹˜‹–›ƒ†Š‡ƒ–‘Ž‘‰‹…ƒŽ’ƒ”ƒ‡–‡”•‹‹Ž‡–‹Žƒ’‹ƒǤ…‘–‘š‹…‘ŽǤ˜‹”‘ǤƒˆǤͳʹͻǡʹͷͲȂ ʹͷ͸Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡…‘‡˜ǤʹͲͳ͸ǤͲ͵ǤͲʹͺ

ǡʹͲͳ͵Ǥ‹”‡…–‹˜‡ʹͲͳ͵Ȁ͵ͻȀ‘ˆ–Š‡‡—”‘’‡ƒ’ƒ”Ž‹ƒ‡–ƒ†‘ˆ–Š‡…‘—…‹Ž‘ˆͳʹ—‰—•–

ʹͲͳ͵ǡ ƒ‡†‹‰ ‹”‡…–‹˜‡• ʹͲͲͲȀ͸ͲȀ ƒ† ʹͲͲͺȀͳͲͷȀ ƒ• ”‡‰ƒ”†• ’”‹‘”‹–›

•—„•–ƒ…‡• ‹ –Š‡ ˆ‹‡Ž† ‘ˆ ™ƒ–‡” ’‘Ž‹…› ȏ ‘…—‡–ȐǤ ˆˆǤ Ǥ —”Ǥ ‹‘Ǥ

Š––’•ǣȀȀ‡—”Ǧ

Ž‡šǤ‡—”‘’ƒǤ‡—Ȁ‡š”‹‡”˜Ȁ‡š”‹‡”˜Ǥ†‘ǫ—”‹αǣǣʹͲͳ͵ǣʹʹ͸ǣͲͲͲͳǣͲͲͳ͹ǣǣ ǡ ʹͲͲͺǤ ‹”‡…–‹˜‡ ʹͲͲͺȀͳͲͷȀ ‘ˆ –Š‡ —”‘’‡ƒ ƒ”Ž‹ƒ‡– ƒ† ‘ˆ –Š‡ ‘—…‹Ž ‘ˆ ͳ͸

‡…‡„‡” ʹͲͲͺ ‘ ‡˜‹”‘‡–ƒŽ “—ƒŽ‹–› •–ƒ†ƒ”†• ‹ –Š‡ ˆ‹‡Ž† ‘ˆ ™ƒ–‡” ’‘Ž‹…›ǡ

ƒ‡†‹‰ ƒ† •—„•‡“—‡–Ž› ”‡’‡ƒŽ‹‰ ‘—…‹Ž ‹”‡…–‹˜‡• ͺʹȀͳ͹͸Ȁǡ ͺ͵Ȁͷͳ͵Ȁǡ ͺͶȀͳͷ͸Ȁǡ ͺͶȀͶͻͳȀǡ ȏ ‘…—‡–ȐǤ ˆˆǤ Ǥ —”Ǥ ‹‘Ǥ †‘‹ǣŠ––’ǣȀȀ‡—”Ǧ

Ž‡šǤ‡—”‘’ƒǤ‡—ȀŽ‡‰ƒŽǦ…‘–‡–ȀȀȀǫ—”‹α…‡Ž‡šǣ͵ʹͲͲͺͲͳͲͷ

ǡʹͲͲʹǤ‹”‡…–‘”ƒ–‡Ǧ‡‡”ƒŽˆ‘”˜‹”‘‡–Ǥ‡ƒ˜›‡–ƒŽ•‹™ƒ•–‡Ǥ ‹ƒŽ”‡’‘”–ȏ

‘…—‡–ȐǤ

Š––’ǣȀȀ‡…Ǥ‡—”‘’ƒǤ‡—Ȁ‡˜‹”‘‡–Ȁ™ƒ•–‡Ȁ•–—†‹‡•Ȁ’†ˆȀŠ‡ƒ˜›̴‡–ƒŽ•”‡’‘”–Ǥ’†ˆ

ȋƒ……‡••‡†͵ǤͳͳǤͳ͹ȌǤ

ǡʹͲͲͲǤ‹”‡…–‹˜‡ʹͲͲͲȀ͸ͲȀ‘ˆ–Š‡—”‘’‡ƒ’ƒ”Ž‹ƒ‡–ƒ†‘ˆ–Š‡‘—…‹Ž‘ˆʹ͵…–‘„‡”

ʹͲͲͲǤ•–ƒ„Ž‹•Š‹‰ƒˆ”ƒ‡™‘”ˆ‘”…‘—‹–›ƒ…–‹‘‹–Š‡ˆ‹‡Ž†‘ˆ™ƒ–‡”’‘Ž‹…›ȏ

‘…—‡–ȐǤˆˆǤǤ—”Ǥ‘—‹–‹‡•Ǥ†‘‹ǣͳͲǤͳͲ͵ͻȀƒ’ͻͺͶʹͳͲͲͳͻ͸

ǡ ʹͲͳͷǤ —”‘’‡ƒ ‘‘† ƒˆ‡–› —–Š‘”‹–›Ǥ …‹‡–‹ˆ‹… •–ƒ–‡‡– ‘ –Š‡ Š‡ƒŽ–ŠǦ„ƒ•‡†

‰—‹†ƒ…‡ ˜ƒŽ—‡• ˆ‘” †‹‘š‹• ƒ† †‹‘š‹ǦŽ‹‡ •Ǥ Ǥ ͳ͵ǡ ͶͳʹͶǤ

†‘‹ǣͳͲǤʹͻͲ͵ȀŒǤ‡ˆ•ƒǤʹͲͳͷǤͶͳʹͶ

Ȁǡ ʹͲͳ͹Ǥ ”ƒ†‡ ƒ† ˆ‘‘† •–ƒ†ƒ”†• ȏ ‘…—‡–ȐǤ

Š––’ǣȀȀ™™™Ǥˆƒ‘Ǥ‘”‰Ȁ͵ȀƒǦ‹͹ͶͲ͹‡Ǥ’†ˆȋƒ……‡••‡†ͳͲǤͳʹǤͳͺȌǤ

ǡ ʹͲͳͺǤ Š‡ •–ƒ–‡ ‘ˆ –Š‡ ™‘”Ž† ˆ‹•Š‡”‹‡• ƒ† ƒ“—ƒ…—Ž–—”‡Ǣ ‡‡–‹‰ –Š‡ •—•–ƒ‹ƒ„Ž‡

†‡˜‡Ž‘’‡–‰‘ƒŽ•Ǥ‘‡Ǥ

ǡʹͲͳ͸ǤŠ‡–ƒ–‡‘ˆ‘”Ž† ‹•Š‡”‹‡•ƒ†“—ƒ…—Ž–—”‡Ǥ‘–”‹„—–‹‰–‘ˆ‘‘†•‡…—”‹–›ƒ†

—–”‹–‹‘ˆ‘”ƒŽŽǤ‘‡Ǥ

ǡʹͲͳͶǤŠ‡•–ƒ–‡‘ˆ™‘”Ž†ˆ‹•Š‡”‹‡•ƒ†ƒ“—ƒ…—Ž–—”‡Ǥ’’‘”–—‹–‹‡•ƒ†…ŠƒŽŽ‡‰‡•Ǥǡ ‘‘†

ƒ†‰”‹…—Ž–—”‡”‰ƒ‹œƒ–‹‘‘ˆ–Š‡‹–‡†ƒ–‹‘•Ǥ‘‡Ǥ

ǡ ʹͲͳͲǤ —Ž–—”‡† ƒ“—ƒ–‹… •’‡…‹‡• ‹ˆ‘”ƒ–‹‘ ’”‘‰”ƒ‡ Ǧ ”‡‘…Š”‘‹• ‹Ž‘–‹…—•

ȋ‹ƒ‡—•ǡ ͳ͹ͷͺȌǤ ˆ‹•Š‡”‹‡• ƒ† ƒ“—ƒ…—Ž–—”‡ ȏ ‘…—‡–ȐǤ

Š––’ǣȀȀ™™™Ǥˆƒ‘Ǥ‘”‰Ȁˆ‹•Š‡”›Ȁ…—Ž–—”‡†•’‡…‹‡•Ȁ”‡‘…Š”‘‹•̴‹Ž‘–‹…—•Ȁ‡ ȋƒ……‡••‡†

ͳʹǤ͵ͳǤͳͺȌǤ

ǡʹͲͲ͸Ǥ ‘‘†•‡…—”‹–›ǡ‘Ž‹…›”‹‡ˆǤ†‘‹ǣͳͲǤͳͲͳ͸Ȁͻ͹ͺǦͲǦͳʹǦ͵ͻͶͺͲ͹Ǧ͸ǤͲͲͲ͵ͻǦ͵

ǡ ʹͲͲ͵Ǥ ƒ–‹‘ƒŽ ƒ“—ƒ…—Ž–—”‡ •‡…–‘” ‘˜‡”˜‹‡™ǣ ‹–‡† ‡’—„Ž‹… ‘ˆ ƒœƒ‹ƒ ȏ

‘…—‡–ȐǤŠ––’ǣȀȀ™™™Ǥˆƒ‘Ǥ‘”‰Ȁˆ‹•Š‡”›Ȁ…‘—–”›•‡…–‘”Ȁƒ•‘̴–ƒœƒ‹ƒȀ‡

ǡ ǡ ǡ ǡǡʹͲͳͺǤŠ‡•–ƒ–‡‘ˆˆ‘‘†•‡…—”‹–›ƒ†—–”‹–‹‘‹–Š‡™‘”Ž†ǡ

Ž‘„ƒŽ ‘‘†•‡…—”‹–›‡’‘”–Ǥ ǡ‘‡Ǥ†‘‹ǣͳͲǤͳͲͻ͵Ȁ…Œ”‡•Ȁ”•–ͲͲ͸

‹ǡǤǡ–—”ǡǤǡ••‡”ǡǤǡ…Š‡––‰‡ǡǤǡ‹ŽŽ‡•ǡǤǡƒ‰ǡǤǡƒ—ǡǤǤǡ”ƒ—•ǡǤǡʹͲͳ͹Ǥ

‡—”‘’•›…Š‘Ž‘‰‹…ƒŽ ‡ˆˆ‡…–• ‘ˆ ‘……—’ƒ–‹‘ƒŽ ‡š’‘•—”‡ –‘ ’‘Ž›…ŠŽ‘”‹ƒ–‡† „‹’Š‡›Ž•Ǥ

‡—”‘–‘š‹…‘Ž‘‰›͸͵ǡͳͲ͸ȂͳͳͻǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡—”‘ǤʹͲͳ͹ǤͲͻǤͲͳͳ

ƒ”…‹ƒǡǤǤǤǡͳͻͻͲǤ ‹•Š‡”‹‡•„‹‘Ž‘‰›‘ˆ‹Žˆ‹•ŠȋChanos chanos ‘”•ƒŽȌǤ

‘‡œ—Ž—ǡǤǤǡ™ƒƒŒ‡ǡǤǡƒ–‹ƒǡǤǤǡʹͲͳͺǤ‡ƒ˜›‡–ƒŽ•ƒ†…›ƒ‹†‡†‹•–”‹„—–‹‘‹–Š‡

˜‹ŽŽƒ‰‡••—””‘—†‹‰—œ™ƒ‰‹‰‘Ž†‹‡‹ƒœƒ‹ƒǤƒœƒ‹ƒǤ…‹ǤͶͶǡͳͲ͹ȂͳʹʹǤ

‘”‡ǡǤǤǡŠƒ’’‡ŽŽǡǤǤǡ ‡–‘ǡǤǤǡ Žƒ™•ǡǤǤǡƒ†ƒŽǡǤǡ”‹•ǡǤǤǡ‘’’ƒ”‹ǡǤǡ‘‡ŽŽ‡”ǡǤǤǡ ʹͲͳͷǤ Ǧʹǣ Š‡ ‡†‘…”‹‡ •‘…‹‡–›ǯ• •‡…‘† •…‹‡–‹ˆ‹… •–ƒ–‡‡– ‘ ‡†‘…”‹‡Ǧ

†‹•”—’–‹‰…Š‡‹…ƒŽ•Ǥ†‘…”Ǥ‡˜Ǥ͵͸ǡͳȂͳͷͲǤ†‘‹ǣͳͲǤͳʹͳͲȀ‡”ǤʹͲͳͷǦͳͲͳͲ

ƒŽŽƒ—‡”ǡǤǡ‡‰ǡǤǡƒ‰ǡǤǦǤǡŠ‡ǡǤǡ•ƒ‹ǡǤǦǤǡ‹—ǡǤǡʹͲͳ͸ǤŠ‡ˆˆ‡…–‘ˆŠ”‘‹…”•‡‹…

š’‘•—”‡‹‡„”ƒˆ‹•ŠǤ‡„”ƒˆ‹•Šͳ͵ǡͶͲͷȂͶͳʹǤ†‘‹ǣͳͲǤͳͲͺͻȀœ‡„ǤʹͲͳ͸Ǥͳʹͷʹ

ƒ‹‰ǡǤǤǡǯ”›ƒǡǤǤǡ”ƒ†ƒŽŽǡǤǤǡ‹…‡ǡǤǤǡʹͲͳʹǤ ‘‘†•ƒˆ‡–›ƒ†ˆ‘‘†•‡…—”‹–›Ǥ

ƒ–Ǥ†—…Ǥ‘™ŽǤ͵ǡͻǤ

ƒ•‡ǡǤǤǤǡ‘Šƒ•‘ǡǤǤǡ–‡’’‡Ž‡”ǡǤǡކ”‹‰ǡǤǡ]•–„›ǡǤǤǡ‡”–•‡ǡǤ Ǥǡ‹‡”ǡ ǤǤǡŽ‡•ƒ†‡”•‡ǡǤǡƒ—Ž•‡ǡǤǤǡ‘’•–ƒ†ǡǤǡʹͲͳͻǤ‹š–—”‡‘ˆ‡”•‹•–‡–”‰ƒ‹…

‘ŽŽ—–ƒ–• ȋ•Ȍ ƒ† œ‘š›‡–Šƒ‡ ȋȌ •Š‘™ ’‘–‡–‹ƒŽ •›‡”‰‹•–‹… ‡ˆˆ‡…–• ‘

‹–‡•–‹ƒŽ–—‘”‹‰‡‡•‹•‹–Š‡Ȁ‹Ȁڏ‘—•‡‘†‡ŽǤŠ‡‘•’Š‡”‡ʹͳͶǡͷ͵ͶȂͷͶʹǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͳͺǤͲͻǤͳʹ͸

ƒ””ƒ†ǡǤǡƒœ”ƒ–‹ǡǤǡ„ƒ””ƒǡǤǡʹͲͲ͸Ǥ‘…‡–”ƒ–‹‘•‘ˆ’‘Ž›…ŠŽ‘”‹ƒ–‡†„‹’Š‡›Ž•‹‹†‘‘”

ƒ‹”ƒ†’‘Ž›„”‘‹ƒ–‡††‹’Š‡›Ž‡–Š‡”•‹‹†‘‘”ƒ‹”ƒ††—•–‹‹”‹‰Šƒǡ‹–‡†

‹‰†‘ǣ ’Ž‹…ƒ–‹‘• ˆ‘” Š—ƒ ‡š’‘•—”‡Ǥ ˜‹”‘Ǥ …‹Ǥ ‡…Š‘ŽǤ ͶͲǡ Ͷ͸͵͵ȂͶ͸͵ͺǤ

†‘‹ǣͳͲǤͳͲʹͳȀ‡•Ͳ͸ͲͻͳͶ͹

ǡʹͲͳͶǤ‹‰ŠŽ‡˜‡Ž’ƒ‡Ž‘ˆ‡š’‡”–•‘ˆ‘‘†•‡…—”‹–›ƒ†—–”‹–‹‘‘ˆ–Š‡…‘‹––‡‡‘

™‘”Ž† ˆ‘‘† •‡…—”‹–›Ǥ —•–ƒ‹ƒ„Ž‡ ˆ‹•Š‡”‹‡• ƒ† ƒ“—ƒ…—Ž–—”‡ ˆ‘” ˆ‘‘† •‡…—”‹–› ƒ†

—–”‹–‹‘Ǥ ƒ‘ͳȂͳͳͻǤ

‘›Ž‡ǡǤǡʹͲͳͷǤ‹‘ƒ……——Žƒ–‹‘‘ˆŠ‡ƒ˜›‡–ƒŽ•ȁ…›…Ž‘’‡†‹ƒǤ…‘ȏ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™Ǥ‡…›…Ž‘’‡†‹ƒǤ…‘Ȁ‡˜‹”‘‡–Ȁ‡…›…Ž‘’‡†‹ƒ•ǦƒŽƒƒ…•Ǧ

–”ƒ•…”‹’–•Ǧƒ†Ǧƒ’•Ȁ„‹‘ƒ……——Žƒ–‹‘ǦŠ‡ƒ˜›Ǧ‡–ƒŽ•ȋƒ……‡••‡†ͳʹǤͳ͹ǤͳͺȌǤ

—‘ǡǤǡ‹ǡǤǡ‹ǡǤǡ—ǡǤǡ‡ƒ‰‡”ǡǤǤǡʹͲͳ͹Ǥ‹•–‘”‹…ƒŽ”‡…‘”†‘ˆ’‘Ž›…ŠŽ‘”‹ƒ–‡†„‹’Š‡›Ž•

ȋ•Ȍƒ†•’‡…‹ƒŽ‘……—””‡…‡‘ˆʹͲͻ‹ƒ•ŠƒŽŽ‘™ˆ”‡•ŠǦ™ƒ–‡”Žƒ‡ˆ”‘‡ƒ•–‡”

Š‹ƒǤŠ‡‘•’Š‡”‡ͳͺͶǡͺ͵ʹȂͺͶͲǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͳ͹ǤͲ͸ǤͲ͹͵

Ž›‹ƒǡǤǡ‘ŠŽƒǡǤǡƒ‡ŽǡǤǡò†‡”ƒǡǤǡʹͲͲ͸Ǥˆƒ–‡ƒ†–”ƒ•’‘”–‘…‡ƒ‘†‡Ž

ˆ‘”’‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–•ƒ†‹–•ƒ’’Ž‹…ƒ–‹‘–‘–Š‡‘”–Š‡ƒǤǤƒ”Ǥ›•–Ǥ͸͵ǡͳȂ ͳͻǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤŒƒ”•›•ǤʹͲͲ͸ǤͲͶǤͲͲ͹

ǡ ʹͲͲͷǤ Š‡ ‹–‡”ƒ–‹‘ƒŽ • ‡Ž‹‹ƒ–‹‘ ‡–™‘”Ǥ ‘–•’‘– ”‡’‘”– ˆ‘” ’ƒ––‡” ‘ˆ

’”ƒ…–‹…‡•᩿ǣ••‘—”…‡•ƒ†”‡Ž‡ƒ•‡•‹ƒœƒ‹ƒǤ

ƒ‹•Šƒƒ”ǡ Ǥǡ •‡–‡ǡ Ǥǡ „ƒŽƒ‰ƒǡ Ǥǡ ƒ–Š‡™ǡ ǤǤǡ ‡‡”‡‰‘™†ƒǡ ǤǤǡ ʹͲͳͶǤ ‘š‹…‹–›ǡ

‡…Šƒ‹• ƒ† Š‡ƒŽ–Š ‡ˆˆ‡…–• ‘ˆ •‘‡ Š‡ƒ˜› ‡–ƒŽ•Ǥ –‡”†‹•…‹’Ǥ ‘š‹…‘ŽǤ ͹ǡ ͸ͲȂ͹ʹǤ

†‘‹ǣͳͲǤʹͶ͹ͺȀ‹–‘šǦʹͲͳͶǦͲͲͲͻ

¡”—’ǡ Ǥǡ ʹͲͲ͵Ǥ ƒœƒ”†• ‘ˆ Š‡ƒ˜› ‡–ƒŽ …‘–ƒ‹ƒ–‹‘Ǥ ”Ǥ ‡†Ǥ —ŽŽǤ ͸ͺǡ ͳ͸͹ȂͳͺʹǤ

†‘‹ǣͳͲǤͳͲͻ͵Ȁ„„ȀŽ†‰Ͳ͵ʹ

ƒ˜‡†ǡǤǡ•ƒ‹ǡǤǡʹͲͳ͹Ǥ‘˜‡”˜‹‡™‘ˆ–Š‡ƒ†˜‡”•‡‡ˆˆ‡…–•‘ˆŠ‡ƒ˜›‡–ƒŽ…‘–ƒ‹ƒ–‹‘

‘ ˆ‹•Š Š‡ƒŽ–ŠǤ ”‘…Ǥ ƒ–ŽǤ …ƒ†Ǥ…‹Ǥ †‹ƒ ‡…–Ǥ ‹‘ŽǤ …‹Ǥ †‘‹ǣͳͲǤͳͲͲ͹Ȁ•ͶͲͲͳͳǦͲͳ͹Ǧ Ͳͺ͹ͷǦ͹

ƒ…Š‘Ž‹ǡǤǤǡƒŠ—ǡǤǡʹͲͳͺǤ‡˜‡Ž•ƒ†Š‡ƒŽ–Š”‹•ƒ••‡••‡–‘ˆŠ‡ƒ˜›‡–ƒŽ•‹•‘‹Žǡ™ƒ–‡”

ƒ†˜‡‰‡–ƒ„Ž‡•‘ˆƒ”‡•ƒŽƒƒǡƒœƒ‹ƒǤǤŠ‡ǤʹͲͳͺǤ†‘‹ǣͳͲǤͳͳͷͷȀʹͲͳͺȀͳͶͲʹ͸͹Ͷ

ƒƒǡ Ǥǡ ƒƒ„‡ǡ Ǥǡ ‹‡•›ǡ ǤǤǡ ƒ–•—ƒ™ƒǡ Ǥǡ ͳͻͻ͹Ǥ ”‰ƒ‘…ŠŽ‘”‹‡ ’‡•–‹…‹†‡• ƒ†

’‘Ž›…ŠŽ‘”‹ƒ–‡† „‹’Š‡›Ž• ‹ ˆ‘‘†•–—ˆˆ• ˆ”‘ •‹ƒ ƒ† ‘…‡ƒ‹… …‘—–”‹‡•Ǥ ‡˜ ˜Ǥ

‘–ƒ‘š‹…‘ŽͳͷʹǡͳȂͷͷǤ

ƒ••‡‰ƒǡǤǤǡƒ–‘ǡǤǤǡʹͲͲͺǤ”•‡‹……‘–ƒ‹ƒ–‹‘Ž‡˜‡Ž•‹†”‹‹‰™ƒ–‡”•‘—”…‡•‹

‹‹‰ ƒ”‡ƒ• ‹ ƒ‡ ‹…–‘”‹ƒ ƒ•‹ ǡ ƒœƒ‹ƒ ǡ ƒ† ‹–• ”‡‘˜ƒŽ —•‹‰ •–ƒ„‹Ž‹œ‡†

ˆ‡””ƒŽ•‘Ž•Ǥ–ǤǤ‹‘ŽǤŠ‡‡‹…ƒŽ…‹Ǥʹǡ͵ͺͻȂͶͲͲǤ

Š‡œ”‹ǡ Ǥǡ ”ƒ•‡”ǡ ǤǤǤǡ ‘—”‹œƒ†‡ŠǦ‹ŽŽƒ„ƒ†‹ǡ Ǥǡ ƒ•–”ƒǡ ǤǤǡ ‡”‰ǡ Ǥǡ ‹‡”ǡ ǤǤǡ

‘’•–ƒ†ǡ Ǥǡ ʹͲͳ͹Ǥ ‹š–—”‡ ‘ˆ ’‡”•‹•–‡– ‘”‰ƒ‹… ’‘ŽŽ—–ƒ–• ƒ†

’‡”ˆŽ—‘”‘‘…–ƒ‡•—Žˆ‘‹…ƒ…‹†‹†—…‡••‹‹Žƒ”„‡Šƒ˜‹‘—”ƒŽ”‡•’‘•‡•ǡ„—–†‹ˆˆ‡”‡–‰‡‡

‡š’”‡••‹‘’”‘ˆ‹Ž‡•‹‡„”ƒˆ‹•ŠŽƒ”˜ƒ‡Ǥ–ǤǤ‘ŽǤ…‹ǤͳͺǤ†‘‹ǣͳͲǤ͵͵ͻͲȀ‹Œ•ͳͺͲʹͲʹͻͳ

‹‰Ǧ‡‹†‡ǡ ǤǤǡ ‡Š–ƒǡ Ǥǡ ‹‘‰ǡ ǤǤǡ ƒŠƒǡ ǤǤǡ –‹‡™‹…œǡ ǤǤǡ ƒ•‡”ǡ Ǥǡ

‡‹†‡ƒǡǤǡ‡–‡”•‘ǡǤǤǡʹͲͳʹǤ‡’”‘†—…–‹˜‡ƒ††‡˜‡Ž‘’‡–ƒŽ–‘š‹…‹–›‘ˆ†‹‘š‹

‹ˆ‹•ŠǤ‘ŽǤ‡ŽŽǤ†‘…”‹‘ŽǤ͵ͷͶǡͳʹͳȂͳ͵ͺǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…‡ǤʹͲͳͳǤͲͻǤͲʹ͹

‹™ƒ‰‘ǡ Ǥǡ ƒ••‹ǡ Ǥǡ ‹ƒ›ƒǡ Ǥǡ ʹͲͳͺǤ ‡•–‹…‹†‡ ”‡•‹†—‡• ‹ ˜‡‰‡–ƒ„Ž‡•ǣ ’”ƒ…–‹…ƒŽ

‹–‡”˜‡–‹‘•–‘‹‹‹œ‡–Š‡”‹•‘ˆŠ—ƒ‡š’‘•—”‡‹ƒœƒ‹ƒǤ—””ǤǤ’’ŽǤ…‹Ǥ

‡…Š‘ŽǤʹ͸ǡͳȂͳͺǤ†‘‹ǣͳͲǤͻ͹͵ͶȀȀʹͲͳͺȀ͵ͺͻ͹͸

‘‘’ƒǦ‡••‡„‘‘ǡǤǡ—‹•ƒǡǤǡ‘‡”•ƒǡǤǤǡ‹††‡”ǡǤǤǤ‡ǡƒƒ—™ǡǤǡ—‹•–”ƒǡ ǤǤǤǤǡƒ—‡’ǡǤǤǤǡͳͻͻͶǤ‹‘š‹ƒ†Ž‡˜‡Ž•‹„Ž‘‘†ƒ†Š—ƒ‹Ž‹”‡Žƒ–‹‘

–‘Ž‹˜‹‰ƒ”‡ƒ•‹Š‡‡–Š‡”Žƒ†•ǤŠ‡‘•’Š‡”‡ʹͻǡʹ͵ʹ͹Ȃʹ͵͵ͺǤ

ƒ —ƒ”†‹ƒǡ ǤǤǡ ƒŽ‡ǡ ǤǤǡ ƒ”˜‡›ǡ Ǥǡ ʹͲͲ͹Ǥ ˜‹†‡…‡ ‘ˆ †‡„”‘‹ƒ–‹‘ ‘ˆ

†‡…ƒ„”‘‘†‹’Š‡›Ž ‡–Š‡” ȋǦʹͲͻȌ ‹ „‹‘–ƒ ˆ”‘ ƒ ™ƒ•–‡™ƒ–‡” ”‡…‡‹˜‹‰ •–”‡ƒǤ

˜‹”‘Ǥ…‹Ǥ‡…Š‘ŽǤͶͳǡ͸͸͸͵Ȃ͸͸͹ͲǤ†‘‹ǣͳͲǤͳͲʹͳȀ‡•Ͳ͹Ͳ͹ʹͺ‰

ƒ†”‹‰ƒǡǤǤǡʹͲͲʹǤŠ‡™‘”Ž†™‹†‡’”‘„Ž‡‘ˆŽ‡ƒ†‹’‡–”‘ŽǤ—ŽŽǤ‘”Ž†‡ƒŽ–Š”‰ƒǤͺͲǡ

͹͸ͺǤ†‘‹ǣͲͲͶʹǦͻ͸ͺ͸ʹͲͲʹͲͲͳͲͲͲͲͲ͵ȏ’‹‹Ȑ

ƒ†”‹‰ƒǡ ǤǤǡ —ŽŽ‡”ǡ Ǥǡ …‘•–ƒǡ ǤǤǤǡ †‡›‹ǡ Ǥǡ ”‘Ž†ǡ Ǥǡ ƒ•—ǡ Ǥ ȋ‹ŽȌǡ ƒŽ†±ǡ ǤǤǡ

‡”–‘ŽŽ‹‹ǡǤǡ‘•‡Ǧǯ‡‹ŽŽ›ǡǤǡ‘—ˆˆ‘”†ǡǤǤǡ”‡›••‡ǡǤǤǡŠ‹Ž‡•ǡǤǡƒŠ‹†‘ŽǡǤǡ‘ŽŽǦ

‡…ǡǤǤǡ”‘’’‡”ǡǤǤǡ ‘„‹ŽǡǤǡ —•–‡”ǡǤǡ”‡‡•–‘‡ǡǤǡƒ‹‡•ǡǤǡƒ”ƒŠƒǡǤǡ

—–‡”ǡǤǡŠƒ”‡ǡǤǡ”—’‹…ǡǤǡƒ’Š‡ƒ”ǡǤǡ‘Šƒ‹ǡǤǡƒ”–‹ǡǤǡƒ–Š‹ƒ•‡ǡǤǡ …‡‡”ǡǤǤǡ—””ƒ›ǡǤǤǤǡ†ƒŠ‹ƒƒŒƒ”ƒǡǤǤǡ‡”‡”ƒǡ Ǥǡ‘–‘«‹ǡǤǡ”‡‡”ǡǤǤǡ

ƒ‡•Šǡ Ǥǡ ‘…•–”ڏǡ Ǥǡ ƒŽ‹ƒ•ǡ Ǥǡ ƒ•‘ǡ ǤǤǡ ƒ†‹Ž›ƒǡ Ǥǡ Ž›ǡǤǤǡ ‹–Šǡ ǤǤǡ

–‡‹‡”ǡ Ǥǡ –‡™ƒ”–ǡ ǤǤǡ —ǡ ǤǤǡ ˜ƒ …Šƒ›…ǡ ǤǤǤǡ ƒ†ƒƒǡ ǤǤǡ —‡ŽŽƒǡ Ǥǡ

Š‘‰ǡ Ǥǡ ʹͲͳ͹Ǥ Š‡ ƒ…‡– ‘‹••‹‘ ‘ ’‘ŽŽ—–‹‘ ƒ† Š‡ƒŽ–ŠǤ ƒ…‡– ͸͹͵͸ǡ ͷͳǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀͲͳͶͲǦ͸͹͵͸ȋͳ͹Ȍ͵ʹ͵ͶͷǦͲ

‡‡ǡǤǦǤǡ‘”†‘ǡǤǤǡƒ–ƒƒ„‡ǡǤͲǤǡͳͻͺ͸Ǥ“—ƒ…—Ž–—”‡‘ˆ‹Žˆ‹•ŠȋChanos chanosȌǣ–ƒ–‡

‘ˆ–Š‡ƒ”–ǤŠ‡…‡ƒ‹…•–‹–—–‡ƒƒ’——ǡƒ™ƒ‹‹ǤǤ

‡‹Œ•ǡǤǤǡ‘’’‡ǡǤǤǡŽ‹‡ǡǤǡƒƒŽ†‡”‡ǡǤǤǤǡ‡‘‘‰–ǡǤǡ‡—••…Š‡”ǡǤǤǡʹͲͲͻǤ

ˆˆ‡…–• ‘ˆ †‹‘š‹•ǡ •ǡ ƒ† • ‘ ‹—‘Ž‘‰› ƒ† Š‡ƒ–‘Ž‘‰› ‹ ƒ†‘Ž‡•…‡–•Ǥ

˜‹”‘Ǥ…‹Ǥ‡…Š‘ŽǤͶ͵ǡ͹ͻͶ͸Ȃ͹ͻͷͳǤ†‘‹ǣͳͲǤͳͲʹͳȀ‡•ͻͲͳͶͺͲˆ

‡‡‹ǡ Ǥǡ ‰‘™‹ǡ Ǥ Ǥǡ ‘†‘ǡ Ǥǡ ʹͲͳ͹Ǥ …—–‡ ’‡•–‹…‹†‡ ’‘‹•‘‹‰ ‹ …Š‹Ž†”‡ǣ ‘•’‹–ƒŽ

”‡˜‹‡™ ‹ •‡Ž‡…–‡† Š‘•’‹–ƒŽ• ‘ˆ ƒœƒ‹ƒǤ Ǥ ‘š‹…‘ŽǤ ʹͲͳ͹ǡ ͺǤ

†‘‹ǣͳͲǤͳͳͷͷȀʹͲͳ͹ȀͶʹͲͺͶͲͷ

‡–‡…Šǡ ͳͻͻͺƒǤ ‹–Š‹— ȋ‹Ȍ Ǧ Š‡‹…ƒŽ ’”‘’‡”–‹‡•ǡ ‡ƒŽ–Š ƒ† ˜‹”‘‡–ƒŽ ‡ˆˆ‡…–•

ȏ ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™ǤŽ‡–‡…ŠǤ…‘Ȁ’‡”‹‘†‹…Ȁ‡Ž‡‡–•ȀŽ‹ǤŠ–

ȋƒ……‡••‡†ͳǤͳǤͳͻȌǤ

‡–‡…Šǡ ͳͻͻͺ„Ǥ ”•‡‹… ȋ•Ȍ Ǧ Š‡‹…ƒŽ ’”‘’‡”–‹‡•ǡ ‡ƒŽ–Š ƒ† ˜‹”‘‡–ƒŽ ‡ˆˆ‡…–•

ȏ ‘…—‡–ȐǤ Š––’•ǣȀȀ™™™ǤŽ‡–‡…ŠǤ…‘Ȁ’‡”‹‘†‹…Ȁ‡Ž‡‡–•Ȁƒ•ǤŠ–

ȋƒ……‡••‡†ͳʹǤͳͳǤͳͺȌǤ

‹‡ǡ ǤǤǤǡ —”•–ǡ Ǥǡ ƒ’’‡ǡ Ǥǡ ʹͲͳͲǤ š’‘•—”‡ ‘ˆ ’‘’—Žƒ–‹‘• –‘ †‹‘š‹• ƒ† ”‡Žƒ–‡†

…‘’‘—†•Ǥ ‘‘†††‹–Ǥ‘–ƒǤͳ͹ǡʹͶͳȂʹͷͻǤ†‘‹ǣͳͲǤͳͲͺͲȀͲʹ͸ͷʹͲ͵ͲͲʹͺ͵͵ʹͶ

‹ƒ”‡•ǡǤǡ‡ŽŽ±•ǡǤǡ‘‹‰‘ǡǤǤǡʹͲͳͷǤ—ƒ‡š’‘•—”‡–‘ƒ†…”‹–‹…ƒŽ‡˜ƒŽ—ƒ–‹‘

‘ˆŠ‡ƒŽ–ŠŠƒœƒ”†•Ǥ”…ŠǤ‘š‹…‘ŽǤͺͻǡ͵͵ͷȂ͵ͷ͸Ǥ†‘‹ǣͳͲǤͳͲͲ͹Ȁ•ͲͲʹͲͶǦͲͳͷǦͳͶͷ͹Ǧͳ

‹‘ǡǤǡ‹Ž†ƒǡͳͻͺͶǤ‹•‡ƒ•‡•‘ˆ‹Žˆ‹•Šǡ‹ǣ—ƒ”‹‘ǡǤǡ‡‹–‡œǡǤȋ†•ǤȌǡ”‘…‡‡†‹‰•‘ˆ–Š‡

‡…‘† –‡”ƒ–‹‘ƒŽ ‹Žˆ‹•Š “—ƒ…—Ž–—”‡ ‘ˆ‡”‡…‡Ǥ •Žƒ† —„Ǥ ‘—•‡ǡ ‡–”‘

ƒ‹Žƒǡ’Ǥ’’ͳͶ͹Ǧͳͷ͵Ǥ

‘ŠƒǡǤǡ”‡‹˜‹ǡǤǡƒ…Š•ǡǤǡ—‹”ǡǤǡʹͲͲ͹ǤŽ‘„ƒŽˆƒ–‡‘ˆ•ǣ—””‡–ƒ†ˆ—–—”‡

”‡•‡ƒ”…Š†‹”‡…–‹‘•Ǥ˜‹”‘Ǥ‘ŽŽ—–ǤͳͷͲǡͳͷͲȂͳ͸ͷǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡˜’‘ŽǤʹͲͲ͹ǤͲ͸ǤͲͷͳ

‘‰‡…‡”ǡ ǤǤǡ ‘‰ƒǡ ǤǤǡ —…‹‡”ǡ Ǥǡ ͳͻͻ͹Ǥ Š‡ Š—ƒ Š‡ƒŽ–Š ‡ˆˆ‡…–• ‘ˆ

ȋ‹…ŠŽ‘”‘†‹’Š‡›Ž–”‹…ŠŽ‘”‘‡–Šƒ‡Ȍ ƒ† • ȋ‘Ž›…ŠŽ‘”‹ƒ–‡† ‹’Š‡›Ž•Ȍ ƒ† ƒ

‘˜‡”˜‹‡™‘ˆ‘”‰ƒ‘…ŠŽ‘”‹‡•‹’—„Ž‹…Š‡ƒŽ–ŠǤ—Ǥ‡˜Ǥ—„Ž‹…‡ƒŽ–ŠͳͺǡʹͳͳȂʹͶͶǤ

†‘‹ǣͳͲǤͳͲͷͷȀ•ǦʹͲͲͺǦͳͲʹ͸ʹͶ͵

‘”„‡”ǡǤǡʹͲͲͺǤš’‘•—”‡‘ˆ‡”‹…ƒ•–‘’‘Ž›„”‘‹ƒ–‡††‹’Š‡›Ž‡–Š‡”•ǤǤš’‘Ǥ…‹Ǥ

˜‹”‘Ǥ’‹†‡‹‘ŽǤͳͺǡʹȂͳͻǤ†‘‹ǣͳͲǤͳͲ͵ͺȀ•ŒǤŒ‡•Ǥ͹ͷͲͲͷ͹ʹ

‘™ǡǤǡ‹‰‰•ǡǤǤǡʹͲͳͷǤ—„Ž‡–ŠƒŽ‡ˆˆ‡…–•‘ˆ…ƒ†‹—‘ƒ—†‹–‘”›•–”—…–—”‡ƒ†ˆ—…–‹‘‹

ˆƒ–Š‡ƒ† ‹‘™• ȋPimephales promelasȌǤ ‹•Š Š›•‹‘ŽǤ ‹‘…Š‡Ǥ Ͷͳǡ ͵ͷ͹Ȃ͵͸ͻǤ

†‘‹ǣͳͲǤͳͲͲ͹Ȁ•ͳͲ͸ͻͷǦͲͳͶǦͻͻͺͺǦ͸

—‹Ž‘ǡǤǡ–ŠƒǡǤǡʹͲͲͻǤ‡ƒ†’‘ŽŽ—–‹‘‹—”„ƒ”‘ƒ†•‹†‡‡˜‹”‘‡–•‘ˆƒ”‡•ƒŽƒƒ

…‹–›Ǥƒœƒ‹ƒǤ…‹Ǥ͵ʹǡ͸ͳȂ͸͹Ǥ†‘‹ǣͳͲǤͶ͵ͳͶȀ–Œ•Ǥ˜͵ʹ‹ʹǤͳͺͶͶͳ

ƒ…Š‹™ƒǡ Ǥ Ǥǡ ʹͲͳͲǤ ‘ƒ•–ƒŽ ƒ”‹‡ ’‘ŽŽ—–‹‘ ‹ ƒ” ‡• ƒŽƒƒ ȋƒœƒ‹ƒȌ ”‡Žƒ–‹˜‡ –‘

”‡…‘‡†‡†‡˜‹”‘‡–ƒŽ“—ƒŽ‹–›–ƒ”‰‡–•ˆ‘”–Š‡‡•–‡”†‹ƒ…‡ƒͻǡͳ͹Ȃ͵ͲǤ

ƒ‰—ǡǤǡ‡”‹‘ǡǤǡƒ”‡”—ǡǤǡŠ‡‰‡ǡǤǡʹͲͳ͸ǤǮ—”†‡•ǯ‘ˆ•‡Ž‡…–‡†Š‡ƒ˜›‡–ƒŽ•‹…‘‘

ˆ‹•Š•’‡…‹‡•ˆ”‘•’‡…‹ˆ‹…‡›ƒˆ”‡•Š™ƒ–‡”•Ǥ–ǤǤ ‹•ŠǤ“—ƒ–Ǥ–—†ǤͶǡͳ͹͵Ȃͳ͹ͻǤ

ƒŠ—”’ƒ™ƒ”ǡ Ǥǡ ʹͲͳͷǤ ˆˆ‡…–• ‘ˆ Š‡ƒ˜› ‡–ƒŽ• ‘ Š—ƒ Š‡ƒŽ–ŠǤ ‹‘‡†Ǥ ‡’‘”–• ͹Ǥ

†‘‹ǣͳͲǤ͵ͺͻʹȀ„”ǤʹͲͳ͸Ǥ͹͸͹

ǡʹͲͳ͸Ǥ‹‹•–”›‘ˆ‰”‹…—Ž–—”‡‹˜‡•–‘…ƒ† ‹•Š‡”‹‡•ǤŠ‡ƒœƒ‹ƒˆ‹•Š‡”‹‡••‡…–‘”ǣ …ŠƒŽŽ‡‰‡•ƒ†‘’’‘”–—‹–‹‡•Ǥƒ”‡••ƒŽƒƒǤ

ƒ›‹Ž‹œ—ǡǤǤǡ†‡‰‡ŽƒǡǤǤǡ‡ŽŽ‡˜‡ǡǤǡŒ‡”˜‡ǡǤǡƒœ™ƒŽƒǡǤǡ‘‰ƒǡǤǡ—ŽŽ‡”ǡǤǤǤǡ

‹‡ǡǤǡ›…Š‡ǡǤǡʹͲͳ͹Ǥ‡ŽˆǦ‡’‘”–‡†•›’–‘•ƒ†’‡•–‹…‹†‡—•‡ƒ‘‰ˆƒ”™‘”‡”•

‹ ”—•Šƒǡ ‘”–Š‡” ƒœƒ‹ƒǣ …”‘•• •‡…–‹‘ƒŽ •–—†›Ǥ ‘š‹…• ͷǡ ͳȂͳ͵Ǥ

†‘‹ǣͳͲǤ͵͵ͻͲȀ–‘š‹…•ͷͲͶͲͲʹͶ

ƒ›‹Ž‹œ—ǡ ǤǤǡ †‡‰‡Žƒǡ ǤǤǡ ƒœ™ƒŽƒǡ Ǥǡ òŽŽ‡”ǡ Ǥǡ ›…Š‡ǡ ǤǤǡ Œ‡”˜‡ǡ Ǥǡ ʹͲͳͷǤ ‡ŽˆǦ

”‡’‘”–‡† Š‡ƒŽ–Š ‡ˆˆ‡…–• ƒ‘‰ •Š‘”– ƒ† Ž‘‰Ǧ–‡” ’‡•–‹…‹†‡ •’”ƒ›‡”• ‹ ”—•Šƒǡ

‘”–Š‡”ƒœƒ‹ƒǣ…”‘•••‡…–‹‘ƒŽ•–—†›Ǥ……—’Ǥ‡†Ǥ‡ƒŽǤˆˆǤ͵Ǥ†‘‹ǣͳͲǤͶͳ͹ʹȀʹ͵ʹͻǦ

͸ͺ͹ͻǤͳͲͲͲʹʹ͵

ƒ”–‹‡œǡ ǤǤǡ •‡‰ǡ Ǥǡ ‡Šǡ Ǥǡ ʹͲͲ͸Ǥ ‹Žˆ‹•Š ȋChanos chanosȌ …—Ž–—”‡᩿ǣ ‹–—ƒ–‹‘• ƒ†

–”‡†•Ǥ‘—”ƒŽǦ ‹•Š‡”‹‡•‘…Ǥƒ‹™ƒǡ͵͵ǡʹʹͻȂʹͶͶǤ

ƒ”—•ŠƒǡǤǡ—ǡǤǡƒ–ƒŽǡǤǡƒ†‹ǡǤǡ…Š™ƒ”–œǡǤǡ‰ǡǤǡ ‡†‹—ǡǤǡ‹Š‘‘˜ǡǤǡŠƒǡ ǤǤǡʹͲͳͺǤŠ‡”‡Žƒ–‹‘•Š‹’„‡–™‡‡’‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–•‡š’‘•—”‡ƒ†–›’‡

ʹ †‹ƒ„‡–‡• ƒ‘‰ ˆ‹”•– ƒ–‹‘• ‹ –ƒ”‹‘ ƒ† ƒ‹–‘„ƒǡ ƒƒ†ƒǣ †‹ˆˆ‡”‡…‡ ‹

†‹ˆˆ‡”‡…‡ ƒƒŽ›•‹•Ǥ –Ǥ Ǥ ˜‹”‘Ǥ ‡•Ǥ —„Ž‹… ‡ƒŽ–Š ͳͷǡ ͳȂͳͻǤ

†‘‹ǣͳͲǤ͵͵ͻͲȀ‹Œ‡”’ŠͳͷͲ͵Ͳͷ͵ͻ

†‡‰‡ŽƒǡǤǤǡ”ƒƒ–Š‡ǡǤǡ‡”‡ƒǡǤǤǡ‘•ŠƒǡǤǤǡƒ†˜‹ǡǤǡƒƒ”‡ǡǤǤǡʹͲͲͻǤ‡ƒ˜›

‡–ƒŽ•ƒ†‘”‰ƒ‘…ŠŽ‘”‹‡”‡•‹†—‡•‹™ƒ–‡”ǡ•‡†‹‡–•ǡƒ†ˆ‹•Š‹ƒ“—ƒ–‹…‡…‘•›•–‡•

‹ —”„ƒ ƒ† ’‡”‹Ǧ—”„ƒ ƒ”‡ƒ• ‹ ƒœƒ‹ƒǤ ƒ–‡”Ǥ ‹”Ǥ ‘‹Ž ‘ŽŽ—–Ǥ ͳȂͳͳǤ

†‘‹ǣͳͲǤͳͲͲ͹Ȁ•ͳͳʹ͹ͲǦͲͲͻǦͲͲͳͻǦ͹

‹”‡”ƒǡǤǤǡʹͲͳͳǤš’‡”‹‡–ƒŽ’‘Ž›…—Ž–—”‡‘ˆ‹Žˆ‹•ŠȋChanos chanosȌƒ†—ŽŽ‡–ȋMugil cephalusȌ—•‹‰‡ƒ”–Š‡’‘†•‹‡›ƒǤ‡•–Ǥ†‹ƒ…‡ƒǤƒ”Ǥ…‹ǤͳͲǡͷͻȂ͹ͳǤ

‘…Š‹ǡ ǤǤǡ ™ƒ†›ƒǡ ǤǤǡ ʹͲͲ͵Ǥ ƒ–‡” —ƒŽ‹–› ‹ –Š‡ –‡‰”ƒ–‡† ƒ”‹…—Ž–—”‡ ‘†

›•–‡•ȋȌƒ–ƒ‘„ƒƒ›ǡƒœ‹„ƒ”ǡƒœƒ‹ƒǤ‡•–Ǥ†‹ƒ…‡ƒǤƒ”Ǥ…‹Ǥʹǡ ͳͷȂ͵ͲǤ

‘†ƒ†—‰—ǡǤǤǡǤ…‘•–ƒǡǤǡʹͲͲͶǤ”‡˜‹‡™‘ˆ‰Ž‘„ƒŽ–‹Žƒ’‹ƒˆƒ”‹‰’”ƒ…–‹…‡•Ǥ“—ƒ…Ǥ•‹ƒ

ͻǡ͹Ȃͳ͸Ǥ†‘‹ǣͳͲǤ͵ͳͲͻȀͻ͹ͺͲʹͲ͵͵ͲͺͻͲͷǦͷ

‘Šƒ‡†Š‡†ǡǤǤǡ ”Þ›•ƒǡǤǤǡƒ”Ž•‡ǡǤǤǡŽƒ•‡”ǡǤǡŽ‹•ƒ„‡–ŠǡǤǡ‡”–•‡ǡǤ Ǥǡ

‡”Šƒ‡‰‡ǡǤǡ‘’•–ƒ†ǡǤǡ‡ŽŽƒǡǤǡ‘•Þ›”ǡǤǡʹͲͳͺǤˆˆ‡…–•‘ˆ†‡ˆ‹‡†‹š–—”‡•

‘ˆ • ƒ† ‡†‘…”‹‡ †‹•”—’–‘”• ‘ –Š‡ •–‡”‘‹† ‡–ƒ„‘Ž‘‡ ‘ˆ –Š‡ Š—ƒ ʹͻͷ

ƒ†”‡‘…‘”–‹…ƒŽ…‡ŽŽŽ‹‡ʹͳͺǡ͵ʹͺȂ͵͵ͻǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͳͺǤͳͳǤͲͷ͹

‘Šƒ‡†ǡǤǤǡʹͲͲʹǤ”‡˜‹‡™‘ˆ™ƒ–‡”“—ƒŽ‹–›ƒ†’‘ŽŽ—–‹‘•–—†‹‡•‹ƒœƒ‹ƒǤ Ǥ—Ǥ˜‹”‘Ǥ͵ͳǡ͸ͳ͹Ȃ͸ʹͳǤ†‘‹ǣͳͲǤͳͷ͹ͻȀͲͲͶͶǦ͹ͶͶ͹Ǧ͵ͳǤ͹Ǥ͸ͳ͹

‘”ƒ‹•ǡǤǡ‘•–ƒǡ ǤǤ‡ǡ‡”‡‹”ƒǡǤ†‡ǤǡʹͲͳʹǤ‡ƒ˜›‡–ƒŽ•ƒ†Š—ƒŠ‡ƒŽ–ŠǤ˜‹”‘Ǥ

‡ƒŽǤǦ‡”‰Ǥ‹••—‡•”ƒ…–Ǥʹʹ͹ȂʹͶͲǤ

‘”‡––‹ǡǤǤǡ—”…Š‹‹ǡǤǤǡ‡ŽŽƒ‰ƒ„ƒǡ Ǥǡƒ’”‹‘ǡ ǤǡʹͲͲ͵Ǥ”ƒ…‡ƒ„‹Ž‹–›‹••—‡•‹ˆ‹•Š‡”›

ƒ† ƒ“—ƒ…—Ž–—”‡ ’”‘†—…–•Ǥ ‡–Ǥ ‡•Ǥ ‘—Ǥ ʹ͹ǡ Ͷͻ͹ȂͷͲͷǤ

†‘‹ǣͳͲǤͳͲʹ͵ȀǣǤͲͲͲͲͲͳͶʹͲ͹ǤͲͳͻͲͲǤͷ…

•—›ƒǡ ǤǤǡ‘…Š‹ǡǤǤǡ‘šǡǤǤǡʹͲͲ͹Ǥ˜‡–‘”›‘ˆ•ƒŽ‡Ǧ•…ƒŽ‡ƒ”‹…—Ž–—”‡‹ƒœƒ‹ƒǤ

—”˜‹˜ƒŽȋ‘†ȌǤ͵͵Ǥ

–—‹ǡǤǡŽ‹‰‘ǡǤǡ—–ƒ›ƒŠ™ƒǡǤǡ‹‰—”ƒǡǤǤǡʹͲͲ͸Ǥ‡‰‡–ƒ–‹‘•–”—…–—”‡ƒ†Š‡ƒ˜›‡–ƒŽ

—’–ƒ‡„›’Žƒ–•‹–Š‡‹‹‰Ǧ‹’ƒ…–‡†ƒ†‘‹‹‰Ǧ‹’ƒ…–‡†•‹–‡•‘ˆ•‘—–Š‡”

Žƒ‡‹…–‘”‹ƒ™‡–Žƒ†•ǤƒœǤǤ…‹Ǥ‘Ž͵ʹǡ͵ͻȂͶͻǤ

òŽŽ‡”ǡǤǤǤǡʹͲͳ͹Ǥ‡”‹ƒ–ƒŽ‡š’‘•—”‡–‘‡˜‹”‘‡–ƒŽ…‘–ƒ‹ƒ–•ƒ†ƒ••‘…‹ƒ–‹‘•–‘

Š‡ƒŽ–Š”‹••‹‘”–Š‡”ƒœƒ‹ƒǤ‘”™‡‰‹ƒ‹˜‡”•‹–›‘ˆ‹ˆ‡…‹‡…‡•Ǥ

òŽŽ‡”ǡǤǤǤǡ‘Ž†‡”ǡǤǡ”›‹Ž†•”—†ǡǤǡ”ސ‡•–ƒ†ǡǤǡƒ”‹‹ǡǤǡ‹‡ǡǤǡƒ›‹Ž‹œ—ǡǤǡ

†‡‰‡ŽƒǡǤǡ‘‹–‹ǡ Ǥǡ—”–ƒ†ŠƒǡǤǡƒ—‡‘‰ƒǡǤǡƒƒ”‡ǡǤǤǡ‘ŽŠƒ—‰ǡǤǡ›…Š‡ǡ ǤǡʹͲͳͺǤ”‡ƒ–ƒŽ‡š’‘•—”‡–‘’‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–•‹‘”–Š‡”ƒœƒ‹ƒƒ†

–Š‡‹” †‹•–”‹„—–‹‘ „‡–™‡‡ „”‡ƒ•– ‹Žǡ ƒ–‡”ƒŽ „Ž‘‘†ǡ ’Žƒ…‡–ƒ ƒ† …‘”† „Ž‘‘†ǡ

˜‹”‘‡–ƒŽ‡•‡ƒ”…ŠǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡˜”‡•ǤʹͲͳͺǤͳʹǤͲʹ͸

òŽŽ‡”ǡǤǤǤǡ‘Ž†‡”ǡǤǡ”›‹Ž†•”—†ǡǤǤǡƒ”‹‹ǡǤǡ‹‡ǡǤǡƒ›‹Ž‹œ—ǡǤǤǡ†‡‰‡ŽƒǡǤǤǡ

‘‹–‹ǡ Ǥǡ —”–ƒ†Šƒǡ Ǥǡ ‘‰ƒǡ ǤǤǡ ƒƒ”‡ǡ ǤǤǡ ›…Š‡ǡ ǤǤǡ ʹͲͳ͹Ǥ ”‰ƒ‘…ŠŽ‘”‹‡

’‡•–‹…‹†‡• ȋ•Ȍ ƒ† ’‘Ž›…ŠŽ‘”‹ƒ–‡† „‹’Š‡›Ž• ȋ•Ȍ ‹ Š—ƒ „”‡ƒ•– ‹Ž ƒ†

ƒ••‘…‹ƒ–‡†Š‡ƒŽ–Š”‹••–‘—”•‹‰‹ˆƒ–•‹‘”–Š‡”ƒœƒ‹ƒǤ˜‹”‘Ǥ‡•ǤͳͷͶǡͶʹͷȂ Ͷ͵ͶǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡˜”‡•ǤʹͲͳ͹ǤͲͳǤͲ͵ͳ

òŽŽ‡”ǡǤǤǤǡ‘Ž†‡”ǡǤǡ”›‹Ž†•”—†ǡǤǤǡ‹‡ǡǤǡލ‡ǡǤǤǡƒ›‹Ž‹œ—ǡǤǤǡ†‡‰‡ŽƒǡǤǤǡ

‘‹–‹ǡ Ǥǡ—”–ƒ†ŠƒǡǤǡ‘‰ƒǡǤǤǡƒƒ”‡ǡǤǤǡ›…Š‡ǡǤǤǡʹͲͳ͸Ǥ”‘‹ƒ–‡†ˆŽƒ‡

”‡–ƒ”†ƒ–• ȋ •Ȍ ‹ „”‡ƒ•– ‹Ž ƒ† ƒ••‘…‹ƒ–‡† Š‡ƒŽ–Š ”‹•• –‘ —”•‹‰ ‹ˆƒ–• ‹

‘”–Š‡”ƒœƒ‹ƒǤ˜‹”‘Ǥ–ǤͺͻȂͻͲǡ͵ͺȂͶ͹Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡˜‹–ǤʹͲͳͷǤͳʹǤͲ͵ʹ

™‡˜—”ƒǡ Ǥǡ –Šƒǡ ǤǤǡ Š‡Š‡ǡ ǤǤǡ ʹͲͲʹǤ ”‰ƒ‘…ŠŽ‘”‹‡ ’‡•–‹…‹†‡ ”‡•‹†—‡• ‹

•‡†‹‡–•ƒ†„‹‘–ƒˆ”‘–Š‡…‘ƒ•–ƒŽƒ”‡ƒ‘ˆƒ”‡•ƒŽƒƒ…‹–›ǡƒœƒ‹ƒǤƒ”Ǥ‘ŽŽ—–Ǥ

—ŽŽǤͶͷǡʹ͸ʹȂʹ͸͹Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀͲͲʹͷǦ͵ʹ͸ȋͲͳȌͲͲ͵͵ͳǦͻ

œ‹”ƒ›ǡǤǡ‹‹”‡‹ǡǤǤǡʹͲͳ͸Ǥ‹‘ƒ……——Žƒ–‹‘‘ˆŠ‡ƒ˜›‡–ƒŽ•‹ƒ”‹‡ˆ‹•Š‡•ȋSiganus sutorǡLethrinus harakǡƒ†Rastrelliger kanagurtaȌˆ”‘ƒ”‡•ƒŽƒƒƒœƒ‹ƒǤ‡‰Ǥ

–—†Ǥƒ”Ǥ…‹Ǥ͹ǡ͹ʹȂͺͲǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ”•ƒǤʹͲͳ͸ǤͲͷǤͲͳͶ

ǡʹͲͳͺǤ‡”ˆ‘”ƒ…‡ƒ—†‹–”‡’‘”–‘–Š‡ƒƒ‰‡‡–‘ˆ’‡•–‹…‹†‡•‹ƒ‰”‹…—Ž–—”‡Ǥƒ”

‡••ƒŽƒƒǤ

Ǧǡ ʹͲͳ͵Ǥ ʹͲͳʹ ‘’—Žƒ–‹‘ ƒ† Š‘—•‹‰ …‡•—•Ǥ ‘’—Žƒ–‹‘ †‹•–”‹„—–‹‘ „›

ƒ†‹‹•–”ƒ–‹˜‡ƒ”‡ƒ•Ǥ

ǡʹͲͳͷǤƒœƒ‹ƒ‹ˆ‹‰—”‡ʹͲͳͶǡŠ‡‹–‡†‡’—„Ž‹…‘ˆƒœƒ‹ƒǡ‹‹•–”›‘ˆ ‹ƒ…‡Ǥ

ƒ”‡•ƒŽƒƒǤ

‹‡—™‘—†–ǡǤǡ—‹ǡǤǤǡ‹‡–‡”•ǡǤǡ‘”†ƒƒǡǤǡ‹••‡”ǡǤǡ›Ž‹ǡǤǡ‘”‰‡ǡǤǤǡ‹‡•›ǡǤǤǡ

‘—™ƒǡǤǡʹͲͲͻǤ‹‘š‹ǦŽ‹‡ …Š‡‹…ƒŽ•‹•‘‹Žƒ†•‡†‹‡– ˆ”‘”‡•‹†‡–‹ƒŽƒ†

‹†—•–”‹ƒŽ ƒ”‡ƒ• ‹ …‡–”ƒŽ ‘—–Š ˆ”‹…ƒǤ Š‡‘•’Š‡”‡ ͹͸ǡ ͹͹ͶȂ͹ͺ͵Ǥ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͲͻǤͲͶǤͲ͸Ͷ

‘‰ƒǡǤǤǡ†‡‰‡ŽƒǡǤǤǡ‹‡ǡǤǡƒ†˜‹ǡǤǡʹͲͳͳǤ••‡••‡–‘ˆˆƒ”‹‰’”ƒ…–‹…‡•ƒ†

—•‡•‘ˆƒ‰”‘…Š‡‹…ƒŽ•‹ƒ‡ƒ›ƒ”ƒ„ƒ•‹ǡƒœƒ‹ƒǤˆ”‹…ƒǤ‰”‹…Ǥ‡•Ǥ͸ǡʹʹͳ͸Ȃ ʹʹ͵ͲǤ

‡”ǡǤǤǡŽ‘„‡‹†ǡǤǤǡ ‘—ƒ†ǡǤǡƒ‰Š‡•–ƒ‹ǡǤǤǡŽǦŽƒ›ƒǡǤǤǡŽƒ‹ǡǤǤǡ‹”ǡǤǡ

ŽǦƒŠƒ••ƒǡ Ǥǡ ʹͲͳʹǤ ƒ†‹— „‹‘ƒ……——Žƒ–‹‘ ƒ† –‘š‹…‹–› ‹ –‹Žƒ’‹ƒ ˆ‹•Š

ȋOreochromis niloticusȌǤ Ǥ ‹Ǥ ‡–Ǥ †˜Ǥ ͳͳǡ ͳ͸ͲͳȂͳ͸Ͳ͸Ǥ

†‘‹ǣͳͲǤ͵ͻʹ͵ȀŒƒ˜ƒƒǤʹͲͳʹǤͳ͸ͲͳǤͳ͸Ͳ͸

‘•–ǡǤ˜ƒ†‡”ǡ‡›‡”ǡǤǡ‡”‡—Ž‡ǡǤǤǤǡʹͲͲ͵Ǥ ‹•Š„‹‘ƒ……——Žƒ–‹‘ƒ†„‹‘ƒ”‡”•‹

‡˜‹”‘‡–ƒŽ ”‹• ƒ••‡••‡–ǣ ƒ ”‡˜‹‡™Ǥ ˜‹”‘Ǥ ‘š‹…‘ŽǤ Šƒ”ƒ…‘ŽǤ ͳ͵ǡ ͷ͹ȂͳͶͻǤ

†‘‹ǣͳͲǤͳͲͳ͸Ȁͳ͵ͺʹǦ͸͸ͺͻȋͲʹȌͲͲͳʹ͸Ǧ͸

ƒ“—‘––‡ǡǤǡƒ‹ŽŽ›ǡǤǡͳͻͻͶǤ˜‹”‘‡–ƒŽ…‘•–”ƒ‹–•ƒ†ƒ“—ƒ…—Ž–—”‡†‡˜‡Ž‘’‡–ǡ‹ǣ

”‘…‡‡†‹‰‘ˆ–Š‡ –‡”ƒ–‹‘ƒŽ‘ˆ‡”‡…‡Ǥ’’Ǥʹ͸ͳȂʹ͹ʹǤ

‡•ƒ–‘”‹ǡǤǤǡ‘•‘‹ǡǤǡ—„ƒ‰‘––‹ǡǤǡ”‹ŽŽ‘ǡǤǡ‡”–ƒœœ‹ǡǤǤǡʹͲͲͻǤƒ…‡”‹…‹†‡…‡‹

–Š‡’‘’—Žƒ–‹‘‡š’‘•‡†–‘†‹‘š‹ƒˆ–‡”–Š‡Dz•‡˜‡•‘ƒ……‹†‡–dzǣ™‡–››‡ƒ”•‘ˆˆ‘ŽŽ‘™Ǧ—’Ǥ

˜‹”‘Ǥ‡ƒŽǤŽ‘„Ǥ……‡••…‹Ǥ‘—”…‡ͺǡͳȂͳͳǤ†‘‹ǣͳͲǤͳͳͺ͸ȀͳͶ͹͸ǦͲ͸ͻǦͺǦ͵ͻ

Ž—ǡǤǤǡ‹ǡǤǡƒŒ‘ǡǤǡʹͲͳͲǤŠ‡‡••‡–‹ƒŽ–‘š‹ǣ’ƒ…–‘ˆœ‹…‘Š—ƒŠ‡ƒŽ–ŠǤ–Ǥ

Ǥ˜‹”‘Ǥ‡•Ǥ—„Ž‹…‡ƒŽ–Š͹ǡͳ͵ͶʹȂͳ͵͸ͷǤ†‘‹ǣͳͲǤ͵͵ͻͲȀ‹Œ‡”’Š͹ͲͶͳ͵Ͷʹ

‘Ž†‡”ǡǤǡƒ„”‹‡Ž•‡ǡǤǤǡ†Žƒ†ǡǤǡƒ˜‹‘˜ƒǡǤǤǡƒ…Š‡˜ǡǤǡލ‡ǡǤǤǡƒƒ”‡ǡǤǤǡ ʹͲͲͺƒǤ ’ƒ–‹ƒŽ ƒ† –‡’‘”ƒŽ …Šƒ‰‡• ‘ˆ …ŠŽ‘”‹ƒ–‡† ’‡•–‹…‹†‡•ǡ •ǡ †‹‘š‹•

ȋ•Ȁ •Ȍƒ†„”‘‹ƒ–‡†ˆŽƒ‡”‡–ƒ”†ƒ–•‹Š—ƒ„”‡ƒ•–‹Žˆ”‘‘”–Š‡”

—••‹ƒǤ…‹Ǥ‘–ƒŽ˜‹”‘Ǥ͵ͻͳǡͶͳȂͷͶǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͲ͹ǤͳͲǤͲͶͷ

‘Ž†‡”ǡ Ǥǡ òŽŽ‡”ǡ ǤǤǡ ”›‹Ž†•”—†ǡ ǤǤǡ †‡ ‘‡”ǡ Ǥǡ ƒ‡”•ǡ Ǥǡ ƒ•–”ƒǡ ǤǤǡ ‹‡ǡ Ǥǡ

†‡‰‡ŽƒǡǤǤǡ‘„‡”‰ǡǤǡ‘‰ƒǡǤǤǡƒ†˜‹ǡǤǡƒƒ”‡ǡǤǤǡ›…Š‡ǡǤǤǡʹͲͳ͸Ǥ‹‘š‹•ǡ

•ǡ …ŠŽ‘”‹ƒ–‡† ’‡•–‹…‹†‡• ƒ† „”‘‹ƒ–‡† ˆŽƒ‡ ”‡–ƒ”†ƒ–• ‹ ˆ”‡‡Ǧ”ƒ‰‡ …Š‹…‡

‡‰‰• ˆ”‘ ’‡”‹Ǧ—”„ƒ ƒ”‡ƒ• ‹ ”—•Šƒǡ ƒœƒ‹ƒǣ ‡˜‡Ž• ƒ† ‹’Ž‹…ƒ–‹‘• ˆ‘” Š—ƒ

Š‡ƒŽ–ŠǤ…‹Ǥ‘–ƒŽ˜‹”‘ǤͷͷͳȂͷͷʹǡ͸ͷ͸Ȃ͸͸͹Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳ͸ǤͲʹǤͲʹͳ

‘Ž†‡”ǡǤǡòŽŽ‡”ǡǤǤǡ›…Š‡ǡǤǤǡ†‡‰‡ŽƒǡǤǤǡ‘‰ƒǡǤǤǡƒ„‹‹ǡ ǤǤǡ„‹•‡ǡǤǤǡƒƒ”‡ǡ ǤǤǡ ƒ†˜‹ǡ Ǥǡ Œ‡”˜‡ǡ Ǥǡ ‹‡ǡ Ǥǡ ʹͲͳͶǤ ‡˜‡Ž• ƒ† ’ƒ––‡”• ‘ˆ ’‡”•‹•–‡– ‘”‰ƒ‹…

’‘ŽŽ—–ƒ–•ȋ•Ȍ‹–‹Žƒ’‹ƒȋ”‡‘…Š”‘‹••’ǤȌˆ”‘ˆ‘—”†‹ˆˆ‡”‡–Žƒ‡•‹ƒœƒ‹ƒǣ

‡‘‰”ƒ’Š‹…ƒŽ†‹ˆˆ‡”‡…‡•ƒ†‹’Ž‹…ƒ–‹‘•ˆ‘”Š—ƒŠ‡ƒŽ–ŠǤ…‹Ǥ‘–ƒŽ˜‹”‘ǤͶͺͺȂ ͶͺͻǡʹͷʹȂʹ͸ͲǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳͶǤͲͶǤͲͺͷ

‘Ž†‡”ǡǤǡƒ˜‹‘˜ƒǡǤǤǡƒ…Š‡˜ǡǤǡލ‡ǡǤǤǡ†Žƒ†ǡǤǤǡƒƒ”‡ǡǤǤǡʹͲͳͲǤ‡˜‡Ž•ƒ†

’ƒ––‡”•‘ˆ‡”•‹•–‡–”‰ƒ‹…‘ŽŽ—–ƒ–•ȋȌ‹•‡Ž‡…–‡†ˆ‘‘†‹–‡•ˆ”‘‘”–Š™‡•–

—••‹ƒȋͳͻͻͺȂʹͲͲʹȌƒ†‹’Ž‹…ƒ–‹‘•ˆ‘”†‹‡–ƒ”›‡š’‘•—”‡Ǥ…‹Ǥ‘–ƒŽ˜‹”‘ǤͶͲͺǡ ͷ͵ͷʹȂͷ͵͸ͳǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳͲǤͲ͹ǤͲ͵͸

‘Ž†‡”ǡǤǡŠ‘•‡ǡǤǡ‹†•–”ڏǡǤǡލ‡ǡǤǤǡƒƒ”‡ǡǤǤǡʹͲͲͺ„Ǥ‡˜‡Ž•ƒ†–‡’‘”ƒŽ

–”‡†• ‘ˆ …ŠŽ‘”‹ƒ–‡† ’‡•–‹…‹†‡•ǡ ’‘Ž›…ŠŽ‘”‹ƒ–‡† „‹’Š‡›Ž• ƒ† „”‘‹ƒ–‡† ˆŽƒ‡

”‡–ƒ”†ƒ–• ‹ ‹†‹˜‹†—ƒŽ Š—ƒ „”‡ƒ•– ‹Ž •ƒ’Ž‡• ˆ”‘ ‘”–Š‡” ƒ† ‘—–Š‡”

‘”™ƒ›ǤŠ‡‘•’Š‡”‡͹͵ǡͳͶȂʹ͵Ǥ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͲͺǤͲ͸ǤͲͲʹ

‘—””‡–ǡǤǡ ƒ—…‘ǡǤǡʹͲͳ͸Ǥ‘„ƒŽ–ǡ‹ǣŠ‹–‡ǡǤǤȋ†ǤȌǡ…›…Ž‘’‡†‹ƒ‘ˆ‡‘…Š‡‹•–”›Ǥ

’”‹‰‡” –‡”ƒ–‹‘ƒŽ —„Ž‹•Š‹‰ǡ ™‹–œ‡”Žƒ†ǡ ’’Ǥ ͳͲȂͳ͵Ǥ †‘‹ǣͳͲǤͳͲͲ͹Ȁͻ͹ͺǦ͵Ǧ͵ͳͻǦ

͵ͻͳͻ͵Ǧͻ

ƒŽ†ïƒǡǤǡÀ‡œǡǤǡƒ›‘ƒǡǤǤǡƒ”…‡Ž×ǡǤǡʹͲͲ͹Ǥ‡”…—”›Ž‡˜‡Ž•ƒ†Ž‹˜‡”’ƒ–Š‘Ž‘‰›‹ˆ‡”ƒŽ

ˆ‹•ŠŽ‹˜‹‰‹–Š‡˜‹…‹‹–›‘ˆƒ‡”…—”›…‡ŽŽ…ŠŽ‘”ǦƒŽƒŽ‹ˆƒ…–‘”›ǤŠ‡‘•’Š‡”‡͸͸ǡͳʹͳ͹Ȃ ͳʹʹͷǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ…Š‡‘•’Š‡”‡ǤʹͲͲ͸ǤͲ͹ǤͲͷ͵

‘–Š—‹•ǡǤǡ—”‡Š‘—–ǡǤǡ—‹ŒǡǤǡ‘‡ǡǤǡ—”ƒ‰™ƒǡǤǡƒ–—œ‹ǡǤǡƒ„ƒ‰ƒ„‡ǡǤǤǡ

Š‘‘ǡ Ǥǡ ʹͲͳͶǤ “—ƒ…—Ž–—”‡ ‹ ƒ•– ˆ”‹…ƒǢ ”‡‰‹‘ƒŽ ƒ’’”‘ƒ…ŠǤ ƒ‰‡‹‰‡ǡ

‡–Š‡”Žƒ†•Ǥ

›Žƒ†‡”ǡ Ǥǡ ƒ‹‰ǡ Ǥǡ ƒ—‰†ƒŠŽǡ Ǥǡ ”‡‹˜‹ǡ Ǥǡ —†ǡ Ǥǡ ʹͲͳͷǤ ‘„‹‹‰ ’Žƒ•ƒ

‡ƒ•—”‡‡–• ƒ† ‡…Šƒ‹•–‹… ‘†‡Ž‹‰ –‘ ‡š’Ž‘”‡ –Š‡ ‡ˆˆ‡…– ‘ˆ • ‘ –›’‡ ʹ

†‹ƒ„‡–‡• ‡ŽŽ‹–—• ‹ ‘”™‡‰‹ƒ ™‘‡Ǥ ˜‹”‘Ǥ ‡•Ǥ ͳͶʹǡ ͵͸ͷȂ͵͹͵Ǥ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ‡˜”‡•ǤʹͲͳͷǤͲ͹ǤͲͲʹ

ƒŽ‡Šǡ Ǥǡ ʹͲͲͺǤ ƒ’–—”‡Ǧ„ƒ•‡† ƒ“—ƒ…—Ž–—”‡ ‘ˆ —ŽŽ‡–• ‹ ‰›’–Ǥ  Ǥ ‘˜ƒ–‡ŽŽ‹ ƒ† Ǥ Ǥ

‘Ž–Š—• ȋ‡†•ȌǤ ƒ’–—”‡Ǧ„ƒ•‡† ƒ“—ƒ…—Ž–—”‡Ǥ Ž‘„ƒŽ ‘˜‡”˜‹‡™ǡ ‹•Š‡”‹‡• ‡…Š‹…ƒŽ

ƒ’‡”Ǥ‘‡Ǥ

ƒ”‹ƒǡ ǤǤǤǡ ʹͲͳ͸Ǥ ••‡••‡– ‘ˆ Š‡ƒŽ–Š ”‹•• ƒ••‘…‹ƒ–‡† ™‹–Š …‘…‡–”ƒ–‹‘• ‘ˆ Š‡ƒ˜›

‡–ƒŽ• ‹ ˆ‹•Š ˆ”‘ –Š‡ …‘ƒ•– ‘ˆ ƒœƒ‹ƒǤ Ǥ —Ž–‹†‹•…‹’Ǥ ‰Ǥ …‹Ǥ –—†Ǥ ʹǡ ͹Ǥ

†‘‹ǣ†‘‹Ǥ‘”‰ȀͳͲǤͶ͵ͳͶȀ‡Œ‡•Ǥ˜ͳͲ‹ͶǤͳʹ

…Š‹––‘—ǡǤǤǡͳͻ͹͵Ǥ”–‹ˆ‹…‹ƒŽ•’ƒ™‹‰ ‘ˆ—ŽŽ‡–ƒ†…—Ž–—”‡‘ˆ—ŽŽ‡–ƒ†‹Žˆ‹•Š‹

ƒ‹™ƒǤ–Ǥ‡–Ǥ“—ƒ…Ǥ‡•Ǥ‡˜ǤͺǤ

Š‘‘ǡǤǡƒ–ƒ‡ǡǤǤǡ‡–‡‰‡”‡ǡǤǡƒŒ‹–ƒ—•ǡǤǤǡ•—›ƒǡ Ǥǡ‘…Š‹ǡǤǡ‰ƒ›ƒǡǤǡ ʹͲͳͳǤ Š‡ •–ƒ–—• ƒ† †‡˜‡Ž‘’‡– ‘ˆ ƒ“—ƒ…—Ž–—”‡ ‹ ƒœƒ‹ƒǡ ƒ•– ˆ”‹…ƒǡ ‹ǣ

–ƒ”ƒŒƒǡǤǡ‘†‹—ǡǤǤǡ‘›‘••ƒǡǤǡ—„‡”‹ǡǤǤǡƒ‹”ǡǤǤǡ‡›‡ŠǡǤǤǡ‰ƒǡǤǡ

‹˜ƒ—ƒ”ǡ ǤǤ ȋ†•ǤȌǡ –‡”ƒ–‹‘ƒŽ ‘ˆ‡”‡…‡ ‘ …‘•›•–‡ ‘•‡”˜ƒ–‹‘ ƒ†

—•–ƒ‹ƒ„Ž‡‡˜‡Ž‘’‡–ȋȌǤ„‘‹˜‡”•‹–›ǡ‘ǡ–Š‹‘’‹ƒǡ’’ǤͺͷȂͻ͹Ǥ

‹‘‹…ŠǡǤǤǡ‹–‡•ǡǤǤǡͳͻͻͷǤŽ‘„ƒŽ†‹•–”‹„—–‹‘‘ˆ’‡”•‹•–‡–‘”‰ƒ‘…ŠŽ‘”‹‡…‘’‘—†•Ǥ …‹‡…‡Ǥʹ͸ͻǡͳͺͷͳȂͳͺͷͶǤ

‹”‘‹•ǡ Ǥǡ ʹͲͲ͵Ǥ –‡”‘‹† ’•›…Š‘•‹•ǣ ”‡˜‹‡™Ǥ ‡Ǥ ‘•’Ǥ •›…Š‹ƒ–”› ʹͷǡ ʹ͹Ȃ͵͵Ǥ

†‘‹ǣͳͲǤͳͲͳ͸ȀͲͳ͸͵Ǧͺ͵Ͷ͵ȋͲʹȌͲͲʹͶͳǦͶ

‘„‘ǡǤǤǡʹͲͳ͵Ǥ…‘‘‹…•—•–ƒ‹ƒ„‹Ž‹–›‘ˆ•ƒŽŽ•…ƒŽ‡‹Žˆ‹•Šˆƒ”‹‰‹ƒœƒ‹ƒǤ‹–‡†

ƒ–‹‘•—‹˜‡”•‹–›ˆ‹•Š‡”‹‡•–”ƒ‹‹‰’”‘‰”ƒ‡Ǥ…‡Žƒ†Ǥ

–ƒ’Ž‡–‘ǡǤǤǡ”ƒœ‹ŽǡǤǡ‘Ž„”‘‘ǡǤǤǡ‹–…Š‡Ž‘”‡ǡǤǤǡ‡‡†‹…–ǡǤǡ‘•–ƒ–‹‘˜ǡǤǡ

‘––‡”ǡǤǡʹͲͲ͸Ǥ˜‹˜‘ƒ†‹˜‹–”‘†‡„”‘‹ƒ–‹‘‘ˆ†‡…ƒ„”‘‘†‹’Š‡›Ž‡–Š‡”ȋ ʹͲͻȌ„›Œ—˜‡‹Ž‡”ƒ‹„‘™–”‘—–ƒ†…‘‘…ƒ”’Ǥ˜‹”‘Ǥ…‹Ǥ‡…Š‘ŽǤͶͲǡͶ͸ͷ͵ȂͶ͸ͷͺǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤƒ’•—•…ǤʹͲͳͳǤͳͲǤͳʹͶ

–‡Ž‡”ǡ Ǥǡ ƒ‡Žǡ Ǥǡ ʹͲͳͲǤ ƒ–Š™ƒ›• ‘ˆ –‘ –Š‡ ”…–‹…ǣ ƒ”‹ƒ„‹Ž‹–‹‡• ƒ†

…‘–”‹„—–‹‘•‘ˆ‘…‡ƒ‹……—””‡–•ƒ†ƒ–‘•’Š‡”‹…–”ƒ•’‘”–ƒ†…Š‡‹•–”›•‘—”…‡•Ǥ

–‘•ǤŠ‡ǤŠ›•ǤͳͲǡͻͻ͸ͷȂͻͻͺͲǤ†‘‹ǣͳͲǤͷͳͻͶȀƒ…’ǦͳͲǦͻͻ͸ͷǦʹͲͳͲ

–‘…Š‘Ž ‘˜‡–‹‘ǡ ʹͲͳͺǤ ‹•–‹‰ ‘ˆ • ‹ –Š‡ –‘…Š‘Ž ‘˜‡–‹‘ ȏ

‘…—‡–ȐǤ

Š––’ǣȀȀ…ŠǤ’‘’•Ǥ‹–ȀŠ‡‘˜‡–‹‘ȀŠ‡•ȀŽŽ•Ȁ–ƒ„‹†ȀʹͷͲͻȀ‡ˆƒ—Ž–Ǥƒ•’š

ȋƒ……‡••‡†͹ǤͳʹǤͳͺȌǤ

–‘…Š‘Ž‘˜‡–‹‘ǡʹͲͲͺǤ˜‡”˜‹‡™Ǥȋ‹…ŠŽ‘”‘Ǧ†‹’Š‡›ŽǦ–”‹…ŠŽ‘”‘‡–Šƒ‡Ȍȏ

‘…—‡–ȐǤ

Š––’ǣȀȀ…ŠǤ’‘’•Ǥ‹–Ȁ’Ž‡‡–ƒ–‹‘Ȁ‡•–‹…‹†‡•ȀȀ˜‡”˜‹‡™Ȁ–ƒ„‹†Ȁ͵͹ͺȀ‡ˆƒ

—Ž–Ǥƒ•’šȋƒ……‡••‡†ͳʹǤͳͺǤͳͺȌǤ

…Š‘—™‘—ǡǤǤǡ‡†Œ‘—ǡǤǤǡƒ–Ž‘ŽŽƒǡǤǤǡ—––‘ǡǤǤǡʹͲͳʹǤ‡ƒ˜›‡–ƒŽ•–‘š‹…‹–›ƒ†–Š‡

‡˜‹”‘‡–ǤͳͲͳǡͳȂ͵ͲǤ†‘‹ǣͳͲǤͳͲͲ͹Ȁͻ͹ͺǦ͵Ǧ͹͸Ͷ͵Ǧͺ͵ͶͲǦͶ

Š‘•‡ǡǤǡ‘Žƒ†‡”ǡǤǡƒƒ‡ǡǤǤǡƒžǡǤǡ ”‘•Šƒ—‰ǡǤǡ‹ƒ‡ǡǤǤǡŠ‘”—†ǡǤǡ‡…Š‡”ǡ Ǥǡ›„‹‰ǡǤǡʹͲͲ͹Ǥ……—’ƒ–‹‘ƒŽ‡š’‘•—”‡–‘Š‡šƒ„”‘‘…›…Ž‘†‘†‡…ƒ‡ƒ–ƒ‹†—•–”‹ƒŽ

’Žƒ–Ǥ˜‹”‘Ǥ…‹Ǥ‡…Š‘ŽǤͶͳǡͷʹͳͲȂͷʹͳ͸Ǥ†‘‹ǣͳͲǤͳͲʹͳȀ‡•Ͳ͹Ͳʹ͸ʹʹ

‘”‡ŽŽǡ Ǥǡ ‘…Š‹ǡ Ǥǡ ʹͲͲ͸Ǥ —”ƒ‰ƒ ‘˜‡”ƒ…‡ ƒ•‡Ž‹‡Ǥ ‘ƒ•–ƒŽ ‡•‘—”…‡• ‡–‡”ǡ

‹˜‡”•‹–›‘ˆŠ‘†‡•Žƒ†Ǥ

”ƒ†‹‰ ƒ† …‘‘‹…•ǡ ʹͲͳͺǤ ƒœƒ‹ƒ ’‘”–• ʹͲͳͺ ȏ ‘…—‡–ȐǤ

Š––’•ǣȀȀ–”ƒ†‹‰‡…‘‘‹…•Ǥ…‘Ȁ–ƒœƒ‹ƒȀ‹’‘”–•ȋƒ……‡••‡†ͳʹǤͳ͹ǤͳͺȌǤ

•—‹‘”‹ǡǤǡ‘—ƒ‰ƒǡǤǡŠ‹„ƒ–ƒǡǤǡ…Š‹ǡǤǡƒƒ›ƒƒǡǤǡ•Š‹ƒ”—ǡǤǡƒƒ‘ǡǤǡƒ‡ǡ Ǥǡ‘•Š‹—”ƒǡǤǡ —”—‡ǡǤǡʹͲͲͺǤ‘‰Ǧ–‡”‡ˆˆ‡…–•‘ˆ’‘Ž›…ŠŽ‘”‹ƒ–‡†„‹’Š‡›Ž•ƒ†

†‹‘š‹• ‘ ’”‡‰ƒ…› ‘—–…‘‡• ‹ ™‘‡ ƒˆˆ‡…–‡† „› –Š‡ —•Š‘ ‹…‹†‡–Ǥ ˜‹”‘Ǥ

‡ƒŽ–Š‡”•’‡…–Ǥͳͳ͸ǡ͸ʹ͸Ȃ͸͵ͲǤ†‘‹ǣͳͲǤͳʹͺͻȀ‡Š’ǤͳͲ͸ͺ͸

ǡͳͻ͹ͻǤ‘˜‡–‹‘‘Ž‘‰Ǧ”ƒ‰‡–”ƒ•„‘—†ƒ”›ƒ‹”’‘ŽŽ—–‹‘Ǥ‡‡˜ƒǡ™‹–œ‡”Žƒ†Ǥ ǡ ǡ ǡ ʹͲͲͻǤ ‡‰‹‘ƒŽ •›–Š‡•‹• ”‡’‘”– ‘ –Š‡ •–ƒ–—• ‘ˆ ’‘ŽŽ—–‹‘ ‹ –Š‡

‡•–‡”†‹ƒ…‡ƒ‡‰‹‘Ǥƒ‹”‘„‹Ǥ

ǡ ǡ ǡ ǡ ͳͻͻͺǤ ˜‡”˜‹‡™ ‘ Žƒ†Ǧ„ƒ•‡† ’‘ŽŽ—–ƒ– •‘—”…‡• ƒ† ƒ…–‹˜‹–‹‡•

ƒˆˆ‡…–‹‰–Š‡ƒ”‹‡ǡ…‘ƒ•–ƒŽǡƒ†ˆ”‡•Š™ƒ–‡”‡˜‹”‘‡–‹–Š‡ƒ•–‡”ˆ”‹…ƒ”‡‰‹‘Ǥ ǡ ʹͲͲ͵Ǥ‡”•‹•–‡– ‘”‰ƒ‹…’‘ŽŽ—–ƒ–• ȋ•ȌǣŠƒ…‹‰…Š‡‹…ƒŽƒƒ‰‡‡– ˆ‘”

’”‘˜‡†”‘†—…–‹˜‹–›ǡƒ”‡–……‡••ƒ†˜‹”‘‡–Ǥ‹‡ƒǡ—•–”‹ƒǤ

ǡʹͲͳ͹ǤʹͲͳ͹Š‹ƒǯ•ˆ‹•Š‡”›ƒ—ƒŽǤŽ‘„ƒŽƒ‰”‹…—Ž–—”ƒŽ‹ˆ‘”ƒ–‹‘‡–™‘”Ǥ

ˆ‘”‡‹‰ƒ‰”‹…—Ž–—”ƒŽ•‡”˜‹…‡‰ƒ‹”‡’‘”–Ǥ

ǡ ʹͲͳ͹Ǥ —ƒ Š‡ƒŽ–Š ”‹• ƒ••‡••‡– ȏ ‘…—‡–ȐǤ

Š––’•ǣȀȀ™™™Ǥ‡’ƒǤ‰‘˜Ȁ”‹•ȀŠ—ƒǦŠ‡ƒŽ–ŠǦ”‹•Ǧƒ••‡••‡–ȋƒ……‡••‡†ͳʹǤͳ͸ǤͳͺȌǤ ǡʹͲͲͷǤ˜‹”‘‡–ƒŽ”‘–‡…–‹‘‰‡…›Ǥ—‹†‡Ž‹‡•ˆ‘”…ƒ”…‹‘‰‡”‹•ƒ••‡••‡–ǡ

Ǥ†‘‹ǣͳͲǤͳͲͺͲȀͳͲͶͲͺͶͶͲ͸ͲͲͻ͹͹͸͹͹

ǡʹͲͲͲǤ—‹†ƒ…‡ˆ‘”ƒ••‡••‹‰…Š‡‹…ƒŽ…‘–ƒ‹ƒ–†ƒ–ƒˆ‘”—•‡‹ˆ‹•Šƒ†˜‹•‘”‹‡•ǣ ‹•Š•ƒ’Ž‹‰ƒ†ƒƒŽ›•‹•Ǥ

ǡͳͻͺ͸Ǥ—‹†‡Ž‹‡•ˆ‘”–Š‡Š‡ƒŽ–Š”‹•ƒ••‡••‡–‘ˆ…Š‡‹…ƒŽ‹š–—”‡•Ǥƒ•Š‹‰–‘ǡ Ǥ†‘‹ǣͳͲǤͳͲͶʹȀʹͲͲͻͲͳʹ͹

ƒŽ‘ǡǤǡ‘””‹•ǡǤǡ”‘‹ǡǤǡʹͲͲͷǤ‡–ƒŽ•ǡ–‘š‹…‹–›ƒ†‘š‹†ƒ–‹˜‡•–”‡••Ǥ—””Ǥ‡†ǤŠ‡Ǥ ͳʹǡͳͳ͸ͳȂͳʹͲͺǤ†‘‹ǣͳͲǤʹͳ͹ͶȀͲͻʹͻͺ͸͹Ͳͷ͵͹͸Ͷ͸͵ͷ

ƒŽ‡”ǡǤǤǡ‹„Ž›ǡǤǤǡ‘’‹ǡǤǤǡ‡ƒƒŽŽǡǤǤǡʹͲͳʹǤ”‹…‹’Ž‡•‘ˆ‡…‘–‘š‹…‘Ž‘‰›ǡ ‘—”–ŠǤ

‡†Ǥ”‡••ǡ‡™‘”Ǥ

ƒ‹ǡǤǤǡ”ƒǡǤǡ•ƒ‹ǡǤǤǡʹͲͳͷǤ‡ƒ†–‘š‹…‹–›᩿ǣƒ”‡˜‹‡™Ǥ–‡”†‹•…‹’Ǥ‘š‹…‘ŽǤͺǡͷͷȂ͸ͶǤ

†‘‹ǣͳͲǤͳͷͳͷȀ‹–‘šǦʹͲͳͷǦͲͲͲͻ

ƒ‹ƒǡ Ǥǡ ʹͲͲ͵Ǥ ••‡••‹‰ –Š‡ ’‘–‡–‹ƒŽ ‘ˆ ’‡”•‹•–‡– ‘”‰ƒ‹… …Š‡‹…ƒŽ• ˆ‘” Ž‘‰Ǧƒ‰‡

–”ƒ•’‘”– ƒ† ƒ……——Žƒ–‹‘ ‹ ’‘Žƒ” ”‡‰‹‘•Ǥ ˜‹”‘Ǥ …‹Ǥ ‡…Š‘ŽǤ ͵͹ǡ ͳ͵ͶͶȂͳ͵ͷͳǤ

†‘‹ǣͳͲǤͳͳͲͻȀǤʹͲͲͺǤͳͶͻ

ƒ‹ƒǡ Ǥǡ ƒ…ƒ›ǡ Ǥǡ ͳͻͻ͵Ǥ Ž‘„ƒŽ ˆ”ƒ…–‹‘ƒ–‹‘ ƒ† …‘Ž† …‘†‡•ƒ–‹‘ ‘ˆ Ž‘™ ˜‘Žƒ–‹Ž‹–›

‘”‰ƒ‘…ŠŽ‘”‹‡…‘’‘—†•‹’‘Žƒ””‡‰‹‘•ǤǤ™‡†‹•Š…ƒ†Ǥ…‹ǤŽ‘„ǤʹʹǡͳͲȂͳͺǤ ǡ ʹͲͳͷǤ ‡ƒ† ‡š’‘•—”‡ ‹ ˆ”‹…ƒ …Š‹Ž†”‡ǣ ‘–‡’‘””› •‘—”…‡• ƒ† …‘…‡”Ǥ ‘—–Š

ˆ”‹…ƒǤ

ǡʹͲͳͳǤŠ‡—•‡‘ˆ‹ƒŽƒ”‹ƒ˜‡…–‘”…‘–”‘ŽǤŽ‘„ƒŽƒŽƒ”‹ƒ”‘‰”ƒ‡Ǥ‡‡˜ƒǤ ǡʹͲͳͲǤ‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–•ǣ‹’ƒ…–‘…Š‹Ž†Š‡ƒŽ–ŠǤ‹Ǥ‡‡–Ǥ͵ͻǡ͸͹Ǥ ǡ ʹͲͲ͹Ǥ ‡ƒŽ–Š ”‹•• ‘ˆ Š‡ƒ˜› ‡–ƒŽ• ˆ”‘ Ž‘‰Ǧ”ƒ‰‡ –”ƒ•„‘—†ƒ”› ƒ‹” ’‘ŽŽ—–‹‘Ǥ

‘’‡Šƒ‰‡ǡ‡ƒ”Ǥ

ǡʹͲͲͳǤ˜‹”‘‡–ƒŽŠ‡ƒŽ–Š…‹–‡”‹ƒʹʹͶǣ”•‡‹…ƒ†ƒ”•‡‹……‘’‘—†•Ǥ

ǡͳͻͻ͹ǤŽ—‹‹—‡‡˜ƒǡ‘”Ž†‡ƒŽ–Š”‰ƒ‹œƒ–‹‘ǡ–‡”ƒ–‹‘ƒŽ”‘‰”ƒ‡‘

Š‡‹…ƒŽƒˆ‡–›ȋ˜‹”‘‡–ƒŽ‡ƒŽ–Š”‹–‡”‹ƒͳͻͶȌǤ‘”Ž†‡ƒŽ–Š”‰ƒ‹œƒ–‹‘Ǥ

—ǡǤǡ‘—ǡǤǡ‹‰ǡǤǡŠ‡ǡǤǡ—ǡǤǡŠ‡ǡǤǡƒǡǤǡƒǡǤǡƒ‰ǡǤǡʹͲͳͶǤ••‘…‹ƒ–‹‘

‘ˆǡƒ†Ȁ „‘†›„—”†‡•™‹–ŠŠ‘”‘‡Ž‡˜‡Ž•ˆ‘”…Š‹Ž†”‡‹ƒ‡Ǧ™ƒ•–‡

†‹•ƒ–Ž‹‰ ƒ”‡ƒ ‘ˆ Š‡Œ‹ƒ‰ ”‘˜‹…‡ǡ Š‹ƒǤ …‹Ǥ ‘–ƒŽ ˜‹”‘Ǥ Ͷͻͻǡ ͷͷȂ͸ͳǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳͶǤͲͺǤͲͷ͹

—ǡǤǡ‘—ǡǤǡ‹‰ǡǤǡŠ‡ǡǤǡ—ǡǤǡŠ‡ǡǤǡƒǡǤǡƒ‰ǡǤǡʹͲͳͷǤˆˆ‡…–•‘ˆ•ƒ†

•‘–Š›”‘‹†Š‘”‘‡ǡŽ›’Š‘…›–‡’”‘Ž‹ˆ‡”ƒ–‹‘ǡŠ‡ƒ–‘Ž‘‰›ƒ†‹†‡›‹Œ—”›

ƒ”‡”• ‹ ”‡•‹†‡–• ‘ˆ ƒ ‡Ǧ™ƒ•–‡ †‹•ƒ–Ž‹‰ ƒ”‡ƒ ‹ Š‡Œ‹ƒ‰ǡ Š‹ƒǤ …‹Ǥ ‘–ƒŽ

˜‹”‘Ǥͷ͵͸ǡʹͳͷȂʹʹʹǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳͷǤͲ͹ǤͲʹͷ

—ǡ Ǥǡ Šƒ‰ǡ Ǥǡ ‹ǡ Ǥǡ …Š”ƒǡ ǤǤǡ ‡––”—’ǡ Ǥǡ ʹͲͲʹǤ †‘…”‹‡ ‡ˆˆ‡…–• ‘ˆ •—„Ž‡–ŠƒŽ

‡š’‘•—”‡–‘’‡”•‹•–‡–‘”‰ƒ‹…’‘ŽŽ—–ƒ–•ȋ•Ȍ‘•‹Ž˜‡”…ƒ”’ȋHypophthalmichthys molitrixȌǤ˜‹”‘Ǥ‘ŽŽ—–ǤͳʹͲǡ͸ͺ͵Ȃ͸ͻͲǤ†‘‹ǣͳͲǤͳͲͳ͸ȀͲʹ͸ͻǦ͹ͶͻͳȋͲʹȌͲͲͳͺʹǦ͵

ƒ•‡‡ǡǤǡ‹”œƒǡǤǤǡŠƒǡǤǤǡƒ—•ƒ”ǡǤǡ‡ŠƒǡǤǡƒ‹ˆǡǤǡʹͲͳ͸Ǥ”ƒ…‡‡–ƒŽ•

Š‡ƒŽ–Š ”‹• ƒ’’”ƒ‹•ƒŽ ‹ ˆ‹•Š •’‡…‹‡• ‘ˆ ”ƒ„‹ƒ ‡ƒǤ ’”‹‰‡”’Ž—• ͷǡ ͺͷͻǤ

†‘‹ǣͳͲǤͳͳͺ͸Ȁ•ͶͲͲ͸ͶǦͲͳ͸ǦʹͶ͵͸Ǧ͸

‡‹–‘—ǡǤǤǡ‡ŠƒƒǡǤǤǤǡʹͲͳͶǤ’ƒ…– ‘ˆ™ƒ–‡”’‘ŽŽ—–‹‘™‹–Š Š‡ƒ˜›‡–ƒŽ•‘ ˆ‹•Š

Š‡ƒŽ–Šǣ ˜‡”˜‹‡™ ƒ† —’†ƒ–‡•Ǥ Ž‘„Ǥ ‡–Ǥ ͳʹǡ ʹͳͻȂʹ͵ͳǤ

†‘‹ǣͳͲǤͷͺʹͻȀ‹†‘•‹Ǥ‰˜ǤʹͲͳͶǤͳʹǤͲʹǤͺʹʹͳͻ

Šƒ‰ǡǤǡŠƒ‘ǡǤǡ—ǡǤǡŠƒ‰ǡǤǡʹͲͳ͹Ǥ‹••‹‘‹˜‡–‘”›ƒ†‡˜‹”‘‡–ƒŽ†‹•–”‹„—–‹‘

‘ˆ †‡…ƒ„”‘‘†‹’Š‡›Ž ‡–Š‡” ‹ Š‹ƒǤ …‹Ǥ ‘–ƒŽ ˜‹”‘Ǥ ͷͻͻȂ͸ͲͲǡ ͳͲ͹͵ȂͳͲͺͳǤ

†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͳ͹ǤͲͷǤͲ͸Ͳ

Š‡‰ǡǤǡƒ‰ǡǤǡŠƒ‰ǡǤǡŠ‡‰ǡǤǡŠƒ‰ǡǤǡŠƒ‰ǡǤǡʹͲͲ͹Ǥ‘’—Žƒ–‹‘Š‡ƒŽ–Š”‹•†—‡

–‘†‹‡–ƒ”›‹–ƒ‡‘ˆŠ‡ƒ˜›‡–ƒŽ•‹–Š‡‹†—•–”‹ƒŽƒ”‡ƒ‘ˆ—Ž—†ƒ‘…‹–›ǡŠ‹ƒǤ…‹Ǥ‘–ƒŽ

˜‹”‘Ǥ͵ͺ͹ǡͻ͸ȂͳͲͶǤ†‘‹ǣͳͲǤͳͲͳ͸ȀŒǤ•…‹–‘–‡˜ǤʹͲͲ͹ǤͲ͹ǤͲͶͶ

APPENDIX

Description of POPs and heavy meals POPs

DDTs: ‹…ŠŽ‘”‘†‹’Š‡›Ž–”‹…ŠŽ‘”‘‡–Šƒ‡ ™ƒ• •›–Š‡•‹œ‡† ‹ ͳͺ͹Ͳ• ƒ† ‹–• ‹•‡…–‹…‹†ƒŽ

’”‘’‡”–‹‡•™‡”‡†‹•…‘˜‡”‡†‹ͳͻ͵Ͳ•„›™‹••„‹‘…Š‡‹•–ƒ—Ž‡”ƒòŽŽ‡”ȋƒŽ‡”‡–

ƒŽǤǡ ʹͲͳʹȌǤ – ™ƒ• ™‹†‡Ž› —•‡† †—”‹‰ ™‘”Ž† ™ƒ” ƒ• ƒ ˜‡…–‘” …‘–”‘Ž ƒ† –Š‡”‡ƒˆ–‡”

‡š–‡•‹˜‡Ž›—•‡†ˆ‘”…‘„ƒ––‹‰ƒŽƒ”‹ƒǡ’‡•–•‹ƒ‰”‹…—Ž–—”‡ƒ†‡…–‘’ƒ”ƒ•‹–‡•‹Ž‹˜‡•–‘…

™‘”Ž†™‹†‡ ȋ–‘…Š‘Ž ‘˜‡–‹‘ǡ ʹͲͲͺǢ ƒŽ‡” ‡– ƒŽǤǡ ʹͲͳʹȌǤ —‡ –‘ ‹–• ’‘–‡–‹ƒŽ –‘š‹…

‡ˆˆ‡…–•–‘™‹Ž†Ž‹ˆ‡ƒ†Š—ƒ•ǡ™ƒ•„ƒ‡†‹ͳͻ͹Ͳ•ƒ†‹…Ž—†‡†‹–Š‡–‘…Š‘Ž

‘˜‡–‹‘‹ʹͲͲͶǤ—‡–‘ƒ„•‡…‡‘ˆ‡“—ƒŽŽ›‡ˆˆ‡…–‹˜‡ƒ†‡…‘‘‹…ƒŽ–‡”ƒ–‹˜‡•‹•

•–‹ŽŽƒŽŽ‘™‡†–‘„‡—•‡†ƒ•˜‡…–‘”…‘–”‘Žƒ•‹†‘‘””‡•‹†—‡•’”ƒ›ȋȌƒ†‹•‡…–‹…‹†‡–”‡ƒ–‡†

‡–•ȋȌ‹ƒŽƒ”‹ƒ‡†‡‹…”‡‰‹‘•‡•’‡…‹ƒŽŽ›‹ˆ”‹…ƒȋǡʹͲͳͳȌǤ‘‡–Š‡Ž‡••ǡ‹ŽŽ‡‰ƒŽ

‘‰‘‹‰—•‡‹ƒ‰”‹…—Ž–—”‡‹••—‰‰‡•–‡†‹•‘‡•‹ƒƒ†ˆ”‹…ƒ…‘—–”‹‡••—…Šƒ•ƒœƒ‹ƒ

ƒ†Š‹ƒǤŠƒ•ˆ‹˜‡‡–ƒ„‘Ž‹–‡•p,p′Ǧǡo,p′Ǧǡp,p′Ǧƒ†p,p′ǦǤp,p′Ǧ‹•

–Š‡’”‹…‹’Ž‡‡–ƒ„‘Ž‹–‡ǡ‹–‹•‘”‡–‘š‹…ƒ†’‡”•‹•–‡––Šƒ–Š‡’ƒ”‡–™‹–Š–Š‡

ŠƒŽˆǦŽ‹ˆ‡‘ˆͳͲ›‡ƒ”•Ǥ

HCHs:‡šƒ…ŠŽ‘”‘…›…Ž‘Š‡šƒ‡•‹•‘‡‘ˆ–Š‡‘•–™‹†‡Ž›—•‡†‹•‡…–‹…‹†‡™‘”Ž†™‹†‡‹•‘‹Ž

ƒ’’Ž‹…ƒ–‹‘ǡ–”‡ƒ–‡–‘ˆ•‡‡†ǡ™‘‘†ˆ‘”’”‡•‡”˜ƒ–‹‘ƒ†Š‘—•‡Š‘Ž†‹•‡…–•Ǥ…‘‡•‹

–™‘ˆ‘”—Žƒ–‹‘•Ǣƒ•ƒ–‡…Š‹…ƒŽ‰”ƒ†‡‹š–—”‡…‘–ƒ‹‹‰ȽǦȋͷͷǦͺͲΨȌǡɀǦȋͺǦͳͷΨȌǡ ȾǦȋͷǦͳͶΨȌǡɁǦȋʹǦͳ͸ΨȌ‹•‘‡”•ƒ†ƒ•’—”‡ɀǦƒŽ•‘‘™ƒ•Ž‹†ƒ‡Ǥ‹†ƒ‡

™ƒ•‹‹–‹ƒŽŽ›—•‡†‹–Š‡ƒ†™‡•–‡”—”‘’‡™Š‹Ž‡–‡…Š‹…ƒŽ™ƒ•—•‡†‹—••‹ƒǡ

‘ƒ‹ƒ †‹ƒ ƒ† Š‹ƒǤ Š‹ƒ •–‹ŽŽ ’”‘†—…‡• –‡…Š‹…ƒŽ ˆ‘” —•‡ ƒ• ‹•‡…–‹…‹†‡ ‹

ƒ‰”‹…—Ž–—”‡ǡ™‘‘†’”‡•‡”˜ƒ–‹‘•ƒ†Š‘‡•ǤȾǦ‹•‘‡”‹•–Š‡‘•–’‡”•‹•–‡–‹•‘‡”ƒ†

‘”‡–‘š‹…–ŠƒŽ‹†ƒ‡ǡ‹–•ŠƒŽˆǦŽ‹ˆ‡‹•ƒ’’”‘š‹ƒ–‡†–‘„‡͹›‡ƒ”•‹Š—ƒǤ

HCBs: ‡šƒ…ŠŽ‘”‘„‡œ‡‡ ‹• ƒ ‘”‰ƒ‘…ŠŽ‘”‹‡ ’‡•–‹…‹†‡ǡ ™Š‹…Š ™ƒ• ‹–”‘†—…‡† ‹

ƒ‰”‹…—Ž–—”‡ƒ•ƒˆ—‰‹…‹†‡ˆ‘”•‡‡†–”‡ƒ–‡–‹ͳͻͶͷǤ–‹•ƒŽ•‘ƒ„›Ǧ’”‘†—…–‹–Š‡’”‘†—…–‹‘

‘ˆŽƒ”‰‡—„‡”‘ˆ…ŠŽ‘”‹ƒ–‡†…‘’‘—†••—…Šƒ•…ƒ”„‘–‡–”ƒ…ŠŽ‘”‹†‡ǡ–”‹…ŠŽ‘”‘‡–Š›Ž‡‡

ƒ†’‡–ƒ…ŠŽ‘”‘„‡œ‡‡ƒ†‡š‹•–•ƒ•ƒ‹’—”‹–›‹•‡˜‡”ƒŽ’‡•–‹…‹†‡ˆ‘”—Žƒ–‹‘•Ǥ–™ƒ•

ƒŽ•‘—•‡†‹–Š‡ƒ—ˆƒ…–—”‡‘ˆˆ‹”‡™‘”•ǡƒ—‹–‹‘ǡƒ†•›–Š‡–‹…”—„„‡”Ǥ–‹•’‘–‡–‹ƒŽŽ›

„‹‘Ǧƒ……——Žƒ–‹˜‡™‹–Š–Š‡ŠƒŽˆǦŽ‹ˆ‡‘ˆ‘”‡–Šƒ‘‡›‡ƒ”‹„‹‘–ƒǤ

CHLs:ŠŽ‘”†ƒ‡•™ƒ•’”‘†—…‡†ƒ•‹‹•‡…–‹…‹†‡•‹ͳͻͶͷƒ†—•‡†‹ƒ‰”‹…—Ž–—”‡ƒ†ˆ‘”

–Š‡…‘–”‘Ž‘ˆ–‡”‹–‡•ǡƒ–•ƒ†…‘…”‘ƒ…Š‡•Ǥ–…‘•‹•–•‘ˆ‘”‡–ŠƒͳʹͲ…‘’‘‡–•„—–

–Š‡ ‘•– …‘‘ ƒ† ‹’‘”–ƒ– ƒ”‡cisǦ…ŠŽ‘”†ƒ‡ǡtransǦ…ŠŽ‘”†ƒ‡ǡtransǦ‘ƒ…ŠŽ‘”ǡ

‘š›…ŠŽ‘”†ƒ‡ƒ†Š‡’–ƒ…ŠŽ‘”Ǥ‹•Š‹‰ŠŽ›’‡”•‹•–‡–™‹–Š–Š‡ŠƒŽˆǦŽ‹ˆ‡‘ˆƒ„‘—–Ͷ›‡ƒ”•Ǥ Mirex:‹”‡š™ƒ•‹–”‘†—…‡†‹ͳͻͷͻƒ†—•‡†ƒ•ƒ‹•‡…–‹…‹†‡ƒ‰ƒ‹•–ƒ–•ƒ†–‡”‹–‡•

ƒ†ˆ‹”‡”‡–ƒ”†ƒ–ˆ‘”’Žƒ•–‹…”—„„‡”ǡ’ƒ‹–•ǡ’ƒ’‡”ƒ†‡Ž‡…–”‹…ƒŽ‡“—‹’‡–•Ǥ‹”‡š‹•Š‹‰ŠŽ›

’‡”•‹•–‡–™‹–Š–Š‡ŠƒŽˆǦŽ‹ˆ‡‘ˆƒ’’”‘š‹ƒ–‡Ž›ͳͲ›‡ƒ”•Ǥ

PCBs: ‘Ž›…ŠŽ‘”‹ƒ–‡† „‹’Š‡›Ž• ƒ”‡ •›–Š‡–‹… …‘’‘—†• …‘•‹•–‹‰ –™‘ „‡œ‡‡ ”‹‰•

Ž‹‡†„›ƒ•‹‰Ž‡…ƒ”„‘–‘…ƒ”„‘„‘†ǤŠ‡„‡œ‡‡”‹‰•Šƒ˜‡•‘‡‘”ƒŽŽ–Š‡Š›†”‘‰‡

ƒ–‘• •—„•–‹–—–‡† „› –Š‡ …ŠŽ‘”‹‡ ƒ–‘•Ǥ Š‡‘”‡–‹…ƒŽŽ› –Š‡”‡ ƒ”‡ ʹͲͻ ‹†‹˜‹†—ƒŽ

…‘‰‡‡”• –Šƒ– …ƒ „‡ ˆ‘”‡† †‡’‡†‹‰ ‘ –Š‡ —„‡” ƒ† ’‘•‹–‹‘ ‘ˆ …ŠŽ‘”‹‡ ƒ–‘•

ȋǡ ʹͲͳͶȌǤ • …ƒ „‡ …ƒ–‡‰‘”‹œ‡† „› †‡‰”‡‡ ‘ˆ …ŠŽ‘”‹ƒ–‹‘Ǥ • ™‹–Š –Š‡ •ƒ‡

—„‡”‘ˆ…ŠŽ‘”‹‡•ƒ”‡–‡”‡†ƒ•ǮŠ‘‘Ž‘‰ǯ•—…Šƒ•ƒ‰”‘—’‘ˆ’‡–ƒǦŠ‡šƒǦƒ†Š‡’–ƒǦ•Ǥ

‡’‡†‹‰ ‘ –Š‡ …ŠŽ‘”‹‡ …‘–‡–ǡ ’”‘†—…–‹‘ ’”‘…‡•• ƒ† ‘”‹‰‹ǡ • ™‡”‡ —•‡† ƒ•

…‘’Ž‡šǡ –‡…Š‹…ƒŽ ‹š–—”‡• •—…Š ƒ• ”‘…Ž‘”ǡ Ž‘’Š‡ǡ Š‡‘…ŠŽ‘”ǡ ƒ‡…ŠŽ‘”ǡ ›”ƒŽ‡‡ǡ ‡…Ž‘”ǡ‡Ž‘”Ǥ•™‡”‡ˆ‹”•–ƒ—ˆƒ…–—”‡†…‘‡”…‹ƒŽŽ›‹ͳͻʹͻƒ††—‡–‘‹–••–ƒ„‹Ž‹–›

ƒ†Š‡ƒ–”‡•‹•–ƒ–ƒ–—”‡™ƒ•—•‡†‹˜ƒ”‹‘—•’”‘†—…–•‹…Ž—†‹‰†‹‡Ž‡…–”‹…ǡ–”ƒ•ˆ‘”‡”•ǡ

Š‡ƒ–‡š…Šƒ‰‡ˆŽ—‹†•ƒ†„—‹Ž†‹‰ƒ–‡”‹ƒŽ•Ǥ‘‡…‘‰‡‡”•™‹–Š‡‹–Š‡”‘‡‘”‘Ž

ƒ–‘‹–Š‡ortho’‘•‹–‹‘ƒ”‡•‹‹Žƒ”–‘†‹‘š‹•‹–‡”•‘ˆ’Š›•‹…‘…Š‡‹…ƒŽ’”‘’‡”–‹‡•Ǥ

Š‡›…ƒ„‹†–Š‡Š”‡…‡’–‘”ȋ ǡʹͲͳͷȌƒ†ƒ”‡–Š‡”‡ˆ‘”‡”‡ˆ‡””‡†–‘ƒ•†‹‘š‹ǦŽ‹‡

•ȋǦ•Ȍǡƒ†Š‡…‡–Š‡‹”–‘š‹…‹–›‹•‡š’”‡••‡†‹–‘š‹…‡“—‹˜ƒŽ‡…›ˆƒ…–‘”•ȋ •ȌǤ

Š‡–‘š‹…‹–›‘ˆ‘–Š‡”…‘‰‡‡”•‹•”‡Žƒ–‡†–‘‘–Š‡”‡…Šƒ‹••ȋǡʹͲͲͲȌǤ PCCDs/PCDFs: ‹‘š‹• ƒ”‡ ‘– ƒ—ˆƒ…–—”‡† „—– ’”‘†—…‡† ƒ• ƒ „›Ǧ’”‘†—…– ‘ˆ ˜ƒ”‹‘—•

‹†—•–”‹ƒŽ’”‘…‡••‡•ȋƒŽ‡”‡–ƒŽǤǡʹͲͳʹȌǤ•Ȁ •ƒ”‡‡š–”‡‡Ž›–‘š‹…–‘Š—ƒ•ǤŠ‡

‡…Šƒ‹•‘ˆ–‘š‹…‹–›‘ˆ†‹‘š‹•‹•ƒ••‘…‹ƒ–‡†™‹–Š–Š‡‹–‡”ƒ…–‹‘™‹–Šƒ…›–‘•‘Ž‹…’”‘–‡‹ǡ

–Š‡ƒ”›ŽŠ›†”‘…ƒ”„‘”‡…‡’–‘”ȋŠȌǡ™Š‹…Š”‡‰—Žƒ–‡•–Š‡•›–Š‡•‹•‘ˆƒ˜ƒ”‹‡–›‘ˆ’”‘–‡‹•Ǥ

Š‡Š”‡…‡’–‘”‹•’”‡•‡–‹ƒ›Š—ƒ–‹••—‡•ǡ‹…Ž—†‹‰–Š‡Ž—‰ǡŽ‹˜‡”ǡ’Žƒ…‡–ƒǡƒ†

Ž›’Š‘…›–‡•Ǥ‡…ƒ—•‡ ‘ˆ–Š‡‹”–‘š‹…‹–›ǡ ‹†‹˜‹†—ƒŽ†‹‘š‹• Šƒ˜‡ „‡‡ ƒ••‹‰‡†ƒ Ǥ Š‡

ˆƒ…–‘”‹•”‡Žƒ–‡†–‘–Š‡‘•––‘š‹…†‹‘š‹…‘’‘—†ǡ–Š‡ʹǡ͵ǡ͹ǡͺǦ–‡–”ƒ…ŠŽ‘”‘†‹„‡œ‘Ǧ’Ǧ†‹‘š‹

™‹–Š–Š‡ ‘ˆͳȋ‘‡ȌǤ

BFRs:”‘‹ƒ–‡†ˆŽƒ‡”‡–ƒ”†ƒ–•Šƒ˜‡„‡‡…‘‡”…‹ƒŽŽ›ƒ—ˆƒ…–—”‡†•‹…‡ͳͻ͸Ͳ•ƒ†

ƒ††‡†‹…‘•—‡”’”‘†—…–•ˆ‘”ˆ‹”‡•ƒˆ‡–›–‘‡Šƒ…‡–Š‡ˆŽƒ‡Ǧ”‡–ƒ”†ƒ…›’”‘’‡”–‹‡•Ǥ‘Ž›

„”‘‹ƒ–‡††‹’Š‡›Ž‡–Š‡”ȋȌǡ‹•ƒŽƒ”‰‡•–‰”‘—’‘ˆ •Ǥ‹‡•ǡ•Šƒ˜‡ʹͲͻ …‘‰‡‡”•ƒ†ƒ”‡…‘’‘—†•‘ˆ–™‘„‡œ‡‡”‹‰•„—–‹…‘–”ƒ”›Ž‹‡†„›ƒ‡–Š‡”ƒ†

•‘‡‘”ƒŽŽ‘ˆ‹–•Š›†”‘‰‡ƒ–‘•ƒ”‡•—„•–‹–—–‡†„›„”‘‹‡ǤŠ‡…‘‡”…‹ƒŽ•™‡”‡

’”‘†—…‡†‹–Š”‡‡‹š–—”‡•’‡–ƒǦȋǦʹͺǡǦͶ͹ǡǦͻͻǡǦͳͲͲȌǡ‘…–ƒǦȋǦͳͷ͵ǡǦͳͷͶǡǦ ͳͺ͵Ȍ ƒ† †‡…ƒǦ ȋǦʹͲ͸ǡ ǦʹͲ͹ǡ ǦʹͲͺǡ ǦʹͲͻȌǤ ‡šƒ„”‘‘…›…Ž‘†‘†‡…ƒ‡ ȋȌ ‹•

ƒ‘–Š‡” ‘•– ™‹†‡Ž› —•‡† ƒ• ƒŽ‹’Šƒ–‹… …›…Ž‹… ƒ††‹–‹˜‡ǡ ‹– ‹• ƒ‹Ž› ƒ• ’‘Ž›•–›”‡‡ ‹

…‘•–”—…–‹‘ƒ–‡”‹ƒŽ••—…Šƒ•–Š‡”ƒŽ‹•—Žƒ–‹‘‘”‘Ž†‡†ˆ‘ƒ’ƒ…‹‰ǡƒ†–‡š–‹Ž‡•ǡ•—…Š

ƒ• Š‘—•‡Š‘Ž† ˆ—”‹–—”‡ ƒ† ƒ’’Ž‹ƒ…‡•Ǥ –Š‡” • ‹…Ž—†‡ ’‡–ƒ„”‘‘–‘Ž—‡‡ ȋȌǡ

’‡–ƒ„”‘‘‡–Š›Ž„‡œ‡‡ ȋȌǡ ‹„”‘‘’”‘’›Ž –”‹„”‘‘’Š‡›Ž ‡–Š‡” ȋȌ ƒ†

Š‡šƒ„”‘‘„‹’Š‡›ŽȋȌǤ

PFASs:‡”ˆŽ—‘”‘ƒŽ›Ž•—„•–ƒ…‡•Šƒ˜‡„‡‡’”‘†—…‡†•‹…‡ͳͻͶͻǡ–Š‡›ƒ”‡…‘’‘—†•‘ˆ

…ƒ”„‘ …Šƒ‹• ™‹–Š ƒ– Ž‡ƒ•– ‘‡ …ƒ”„‘ ˆ—ŽŽ› „‘†‡† –‘ ˆŽ—‘”‹‡ ƒ–‘•Ǥ Š‡› ƒ”‡ Š‹‰ŠŽ›

’‡”•‹•–‡–Ž‹‡‘–Š‡”•ǡŠ‘™‡˜‡”–Š‡›ƒ”‡Ž‡••Ž‹’‘’Š‹Ž‹…ǤŠ‡›Šƒ˜‡„‡‡—•‡†ƒ•”‡’‡ŽŽ‡–

‹˜ƒ”‹‘—•…‘•—‡”’”‘†—…–•ǡˆ‘‘†„‘š‡•ǡ‘—–†‘‘”…Ž‘–Š‹‰ǡ‘”‡Ǧ‡šǡ•‹™ƒšǡˆ—”‹–—”‡Ǥ

Š‡‹…ƒŽ•–”—…–—”‡•‘ˆ•‹…Ž—†‡†‹–Š‹••–—†›

DDT PCB

DDE PCDD

y-HCH PBDE

HCB HBCDD

CHL PFAS

Mirex HBB

Heavy metals

Fe: ”‘ ‹• ‡••‡–‹ƒŽ ˆ‘” Šƒ‡‘‰Ž‘„‹ ˆ‘”ƒ–‹‘ ȋƒ•‡‡ ‡– ƒŽǤǡ ʹͲͳ͸Ȍ ”‡•’‘•‹„Ž‡ ˆ‘”

‘š›‰‡–”ƒ•’‘”–ƒ–‹‘‹–Š‡„‘†‹‡•‘ˆŽ‹˜‹‰‘”‰ƒ‹•ȋƒ”‹ƒǡʹͲͳ͸Ȍƒ†‹’‘”–ƒ–ˆ‘”ƒ›

˜‹–ƒŽ’”‘–‡‹•ƒ†‡œ›‡•‹Ž‹˜‹‰‘”‰ƒ‹••ȋƒ‹•Šƒƒ”‡–ƒŽǤǡʹͲͳͶǢƒŽ‘‡–ƒŽǤǡʹͲͲͷȌǤ Co: ‘„ƒŽ– ‹• ‡••‡–‹ƒŽ ‹ Š—ƒ ƒ• ’ƒ”– ‘ˆ ˜‹–ƒ‹ ͳʹǤ – Šƒ• „‡‡ —•‡† ‹ –‡…Š‘Ž‘‰›

‹†—•–”‹‡••—…Šƒ•”‡…Šƒ”‰‡ƒ„Ž‡„ƒ––‡”‹‡•ǡƒŽŽ‘›ƒ†…ƒ–ƒŽ›•–ȋ‘—””‡–ƒ† ƒ—…‘ǡʹͲͳ͸ȌǤ

Cu:‘’’‡”‹•‡••‡–‹ƒŽˆ‘”‡–ƒ„‘Ž‹…ƒ…–‹˜‹–‹‡•‹–Š‡„‘†›‘ˆŽ‹˜‹‰‘”‰ƒ‹••Ǥ–‹•—•‡†‹

–Š‡ ‡œ›ƒ–‹… ’”‘…‡•• ‹ –Š‡ ’”‘†—…–‹‘ ‘ˆ Šƒ‡‘‰Ž‘„‹ ȋƒ‰— ‡– ƒŽǤǡ ʹͲͳ͸ȌǤ – ‹•

•—’’Ž‡‡–‡†‹–Š‡†‹‡–‘ˆˆƒ”‡†ˆ‹•ŠȋǡʹͲͲ͹ȌǤ

Cr:Š”‘‹—‹•‡••‡–‹ƒŽˆ‘”–Š‡‡–ƒ„‘Ž‹•‘ˆ…ƒ”„‘Š›†”ƒ–‡ȋ…Š‘—™‘—‡–ƒŽǤǡʹͲͳʹȌǤŠ‡

Ž‡˜‡Ž•‘ˆ”‹–Š‡‡˜‹”‘‡–•Šƒ˜‡„‡‡‹…”‡ƒ•‡†„›˜ƒ”‹‘—•ƒ–Š”‘’‘‰‡‹…ƒ…–‹˜‹–‹‡•

•—…Š ƒ• Ž‡ƒ–Š‡” ƒ† –‡š–‹Ž‡ǡ †›‹‰ ƒ† ’”‹–‹‰ ‹†—•–”‹‡• ƒ† …Š”‘ƒ–‡ ’”‘†—…–‹‘

ȋ—–Šƒǡ ʹͲͳͷǢ …Š‘—™‘— ‡– ƒŽǤǡ ʹͲͳʹȌǤ ” Šƒ• „‡‡ —•‡† ƒ• ™‘‘† ’”‡•‡”˜ƒ–‹˜‡• ƒ†

ˆ—‰‹…‹†‡•ȋ†‡‰‡Žƒ‡–ƒŽǤǡʹͲͲͻȌ‹‡Ž‡…–”‘’Žƒ–‹‰ƒ†ˆ‡”–‹Ž‹œ‡”•ƒ†—•‡†‹’—Ž’ƒ†’ƒ’‡”

’”‘†—…–‹‘ȋƒ‹•Šƒƒ”‡–ƒŽǤǡʹͲͳͶȌǤ

Zn: ‹… ‹• ƒ ‹’‘”–ƒ– –”ƒ…‡ ‡Ž‡‡– ƒ† ‹…”‘—–”‹‡– –Šƒ– Šƒ• ƒ •‹‰‹ˆ‹…ƒ– ”‘Ž‡ ‹

‡œ›ƒ–‹…ƒ…–‹‘•ǡ‹—‡•›•–‡ƒ†‡—”‘–”ƒ•‹••‹‘ȋ—–ŠƒǡʹͲͳͷȌǤ

Ni: ‹…‡Ž ‹• ”‡Ž‡ƒ•‡† ‹ –‘ –Š‡ ‡˜‹”‘‡– „› „‘–Š ƒ–—”ƒŽ ƒ† ƒǦƒ†‡ •‘—”…‡•

ȋ—–ŠƒǡʹͲͳͷȌǤ‹‹•‡••‡–‹ƒŽ–‘ƒ›‘”‰ƒ‹••ƒ–Ž‘™…‘…‡–”ƒ–‹‘ƒ†„‡…‘‡•–‘š‹…

™Š‡‹–•…‘…‡–”ƒ–‹‘‡š…‡‡†•–Š”‡•Š‘Ž†Ž‡˜‡Ž•Ǥ

Li:‹–Š‹—‹•ƒƒŽƒŽ‹‡–ƒŽǡ™Š‹…Š‡š‹•–ƒ–—”ƒŽŽ›ƒ•‹š–—”‡‘ˆ‹•‘–‘’‡•Ǥ‹–Š‹—‹•—•‡†‹

˜ƒ”‹‘—• ‹†—•–”‹ƒŽ ƒ’’Ž‹…ƒ–‹‘ •—…Š ƒ• Ž‹–Š‹— „ƒ––‡”‹‡• ƒ† ƒ‹” …‘†‹–‹‘‡”• ȋ‡–‡…Šǡ ͳͻͻͺƒȌǤ–Šƒ•ƒŽ•‘„‡‡—•‡†‹˜ƒ”‹‘—•‡†‹…ƒ–‹‘ˆ‘”’•›…Š‹ƒ–”‹……‘†‹–‹‘••—…Šƒ•„‹’‘Žƒ”

†‹•‘”†‡”•ȋ‹”‘‹•ǡʹͲͲ͵ȌǤ

Se:‡Ž‡‹—‹•ƒ‘‰–Š‡”ƒ”‡‡Ž‡‡–•‘‡ƒ”–Š•ƒ†‹•‘•–Ž›ˆ‘—†‹‰”‘—†ƒ†•—”ˆƒ…‡

™ƒ–‡”•Ǥ–‹•ƒ‡••‡–‹ƒŽ‡Ž‡‡–ˆ‘”‘”ƒŽ‰”‘™–Šƒ†’Š›•‹‘Ž‘‰‹…ƒŽˆ—…–‹‘•‘ˆƒ‹ƒŽ•

•—…Šƒ•–Š›”‘‹†‡œ›‡‡–ƒ„‘Ž‹•‹•’‡”ƒ–‘‰‡‡•‹•ȋ—–ŠƒǡʹͲͳͷȌǤ

V:ƒƒ†‹—‹•ˆ‘—†ƒ–—”ƒŽŽ›ƒ†‹•”‡Ž‡ƒ•‡†„›‹†—•–”‹ƒŽ‡‹••‹‘•–Š”‘—‰Š…‘ƒŽƒ†ˆ—‡Ž

„—”‹‰ ȋ—–Šƒǡ ʹͲͳͷȌǤ – ‹• ‡••‡–‹ƒŽ ‹ ”‡‰—Žƒ–‹‰ ˜ƒ”‹‘—• ‡œ›ƒ–‹… ƒ…–‹˜‹–‹‡• ˆ‘”

‡–ƒ„‘Ž‹……Šƒ‰‡•Ǥ

Pb:‡ƒ†Šƒ•‘‹’‘”–ƒ–„‹‘Ž‘‰‹…ƒŽ”‘Ž‡‹–Š‡„‘†›‘ˆŽ‹˜‹‰‘”‰ƒ‹••Ǥ–‹•ƒ‘‰–Š‡

‘•––‘š‹…Š‡ƒ˜›‡–ƒŽ•ˆ‘”Ž‹˜‹‰‘”‰ƒ‹••‹…Ž—†‹‰ˆ‹•Šƒ†Š—ƒȋ…Š‘—™‘—‡–ƒŽǤǡ ʹͲͳʹȌǤ–•Ž‡˜‡Ž•Šƒ•„‡‡‹…”‡ƒ•‡†„›˜ƒ”‹‘—•ƒ–Š”‘’‘‰‡‹…ƒ…–‹˜‹–‹‡•‹…Ž—†‹‰„—”‹‰‘ˆ

ˆ—‡Žǡ ‹‹‰ǡ ƒ† ƒ—ˆƒ…–—”‡ ‘ˆ †‹ˆˆ‡”‡– …‘‘†‹–‹‡• •—…Š ƒ• ’ƒ‹–•ǡ „ƒ––‡”‹‡•ǡ

ƒ—‹–‹‘•ȋ…Š‘—™‘—‡–ƒŽǤǡʹͲͳʹȌƒ†—•‡‘ˆ„…‘–ƒ‹‹‰ˆ—‡Žȋ—‹Ž‘ƒ†–Šƒǡ ʹͲͲͻȌǤ

Hg:‡”…—”›‹•‘‡‘ˆ–Š‡‘•––‘š‹…‡–ƒŽ•–Šƒ–‹•™‹†‡•’”‡ƒ†‹–Š‡‡˜‹”‘‡–Ǥ‰Ž‡˜‡Ž•

ƒ”‡‹…”‡ƒ•‹‰‹–Š‡‡˜‹”‘‡–†—‡–‘—•‡‹ˆ—‰‹…‹†‡•…Š‡‹…ƒŽ•ƒ†˜ƒ”‹‘—•‹†—•–”‹ƒŽ

ƒ…–‹˜‹–‹‡•ȋǡͳͻͻͻǢ—–ŠƒǡʹͲͳͷǢ…Š‘—™‘—‡–ƒŽǤǡʹͲͳʹȌƒ†‹‹‰ȋ‘•…Š‡–ƒŽǤǡ ʹͲͳ͸ȌǤ —”‹‰ ‘ˆ ˆ‘••‹Ž ˆ—‡Ž ƒ† •‡Ž–‹‰ ‘” ‡–ƒŽ ‘”‡• …‘–”‹„—–‡• •‹‰‹ˆ‹…ƒ–Ž› –‘

ƒ–‘•’Š‡”‹…‰Ǥ

Cd:ƒ†‹—‹•ƒƒ–—”ƒŽŽ›‘……—””‹‰‡–ƒŽǤ–‹•—•‡†‹˜ƒ”‹‘—•‹†—•–”‹ƒŽ’”‘†—…–‹‘••—…Š

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ERRATA

Errata

Page Line Original text Corrected text

7 19-20 A study by (Al-sawafi et al., 2017) A study by Al-sawafi et al., (2017) 7 21 (Low and Higgs, 2015) Low and Higgs, (2015)

8 17 paragraph subchapters

9 11 EF

ED

EF

ED

9 12 FIR FIR

9 13 RFD RfD

9 14 WAB WAB

9 14 TA TA

9 15 EF*ED EF*ED

11 2 EQS EQSBIOTA

26 22 95^th EWI for DL-PCBs PCB-118 95^th percentile EWI for DL-PCBs: PCB-118

27 11 to give when to give a call when

28 4 concerted concentrated

29 4 in Tanzania DDTs in Tanzania. DDTs

32 5 in all sites (paper II). in all sites.

32 16 (USEPA, 1986) USEPA, (1986)

Appendix Heavy metals Fe

organism (Saria, 2016). and organism (Saria, 2016) and Appendix

Heavy metals As

and inhuman and in human

PAPERS I-III

Occurrence and levels of persistent organic pollutants (POPs) in farmed and wild marine fish from Tanzania. A pilot study

Eliezer Brown Mwakalapa^a,b,d, Aviti John Mmochi^b, Mette Helen Bjorge Müller^a, Robinson Hammerthon Mdegela^c, Jan Ludvig Lyche^a, Anuschka Polder^a,*

aDepartment of Food Safety and Infection Biology, Norwegian University of Life Sciences, P. O. Box 8146 Dep, N-0033 Oslo, Norway bInstitute of Marine Sciences, University of Dar es Salaam, P. O. Box 668, Mizingani Road, Zanzibar, Tanzania

cDepartment of Veterinary Medicine and Public Health, Sokoine University of Agriculture, P. O. Box 3021, Morogoro, Tanzania dDepartment of Health Sciences and Technology, Mbeya University of Science and Technology, P. O. Box 131, Mbeya, Tanzania

h i g h l i g h t s g r a p h i c a l a b s t r a c t

DDTs were the major POPs in farmed and wild milkfish and mullets.

p,p⁰-DDE was 572 times higher in wild milkfish than in farmed milkfish from Mtwara.

PCBs and PBDEs were low and in varying ranges in milkfish and mullets.

HBCDD in mullet from Pemba war-rant further research on its occur-rence in the region.

a r t i c l e i n f o

Article history:

Received 11 June 2017 Received in revised form 23 September 2017 Accepted 25 September 2017 Available online 26 September 2017 Handling Editor: Myrto Petreas

Keywords:

Persistent organic pollutants (POPs) DDTs

In 2016, farmed and wild milkfish (Chanos chanos) and mullet (Mugil cephalus) from Tanzania mainland (Mtwara) and Zanzibar islands (Pemba and Unguja) were collected for analyses of persistent organic pollutants (POPs). Fish livers were analysed for organochlorine pesticides (OCPs), polychlorinated bi-phenyls (PCBs), brominatedflame retardants (BFRs). Muscle tissue was used for analyses of per-fluoroalkyl substances (PFASs). The major contaminant wasp,p⁰-DDE. The highestp,p⁰-DDE concentration was found in wild milkfish from Mtwara (715.27 ng/g lipid weight (lw)). This was 572 times higher than the maximum level detected in farmed milkfish from the same area. The ratios ofp,p⁰-DDE/p,p⁰-DDT in wild milkfish and mullet from Mtwara and Pemba indicate historical use of DDT. In contrast, ratios in farmed milkfish from Unguja and Mtwara, suggest recent use. The levels of HCB, HCHs and trans-non-achlor were low.P

10PCBs levels were low, ranging from<LOD to 8.13 ng/g lw with the highest mean level found in farmed milkfish from Shakani, Unguja (3.94 ng/g lw). The PCB pattern was dominated by PCB -153>-180>-138. PBDEs were detected in low and varying levels in all locations. BDE-47 was the dominating congener, and the highest level was found in farmed milkfish from Jozani (1.55 ng/g lw).

HBCDD was only detected in wild mullet from Pemba at a level of 16.93 ng/g lw. PFAS was not detected in any of the samples. POP levels differed between geographic areas and between farmed and wildfish.

Human activities seem to influence levels on PCBs and PBDEs on Unguja.

©2017 Elsevier Ltd. All rights reserved.

*Corresponding author. Norwegian University of Life Sciences, Campus Adam-stuen, P.O. Box 8146 Dep, N-0033 Oslo, Norway.

E-mail address:anuschka.polder@nmbu.no(A. Polder).

Contents lists available atScienceDirect

Chemosphere

j o u r n a l h o me p a g e :w w w . e l s e v i e r . c o m/ l o c a t e / c h e m o s p h e r e Chemosphere 191 (2018) 438e449

1. Introduction

Persistent Organic Pollutants (POPs) are halogenated chemical substances that are characterised by high lipophilicity and chemical persistence. POPs accumulate in fatty tissues and biomagnify in the food chain (Lohmann et al., 2007). They are volatile and may un-dergo long range atmospheric transport and be found far from where they were used or manufactured (Polder et al., 2014; Wania and Mackay, 1993). Since 1940s POPs have been manufactured and used in a wide range of products such as pesticides, transformer oil, building materials,flame retardants, antifouling agents and cool-ants. They have also been unintentionally released as by-products in combustion processes (UNIDO, 2003). In aquatic ecosystems, POPs bioaccumulate in organic matter and aquatic organisms including fish (Walker et al., 2012). POPs, such as dichlor-odiphenyltrichloroethane and metabolites (DDTs), polychlorinated biphenyls (PCBs) and brominatedflame retardants (BFRs), have been documented to cause adverse health effects in animals and humans (Trollerud, 2013; Walker et al., 2012), such as egg shell thinning in marine birds (Bouwman et al., 2008; Ratcliffe, 1970), reproductive impairment in seals (Bergman, 2007) in whales (Beland et al., 1993) and endocrine disruption infish (Berg et al., 2016). The Stockholm Convention, a global treaty for protecting humans and the environment against POPs contamination, has listed more than 20 POPs so far (Stockholm Convention, 2016).

Tanzania ratified the Convention in 2004 and has a national implementation plan. Due to the hot and humid tropical climate in Tanzania, many pests and weeds threaten food production in agriculture. Thus, use of pesticides is inevitable. Several studies have documented the presence of POPs in the Tanzanian environ-ment, including food and humans, due to discharges from agri-cultural activities, malaria control, waste disposals and obsolete stockpiles (Kariathi et al., 2016; Kishimba et al., 2004; Lema et al., 2014; Machiwa, 2010; Mtashobya and Nyambo, 2014; Müller et al., 2016, 2017; Mwevura, 2014; Nonga et al., 2011; Polder et al., 2014, 2016).

Fish represents a valuable source of proteins and nutrients for the general population and is of high importance for food security and economy in many countries (FAO, 2016). Therefore, aquacul-ture is rapidly expanding worldwide (FAO, 2014) including Tanzania, (Watengere et al., 2008). The main farmedfish species in Tanzania are Nile tilapia (Oreochromis niloticus), African catfish (Clarius gariepinus), rainbow trout (Oncorhynchus mykiss) in fresh water, and milkfish (Chanos chanos) and mullet (Mugil cephalus) in marine waters (Rothuis et al., 2014). Whereas a few studies on POPs have been done in fresh water and marinefishes in Tanzania (Machiwa, 2010; Mdegela et al., 2009; Mwevura et al., 2002; Polder et al., 2014), no studies have been performed on POPs levels in farmedfish in Tanzania in comparison to POP levels in corre-sponding wild species.

The main objective of the present study was to assess the levels and occurrence of POPs in farmed and wild milkfish and mullet from Jozani and Shankani at Unguja, Mtwara, and Pemba Islands in Tanzania.

2. Materials and methods

2.1. Description of species and study sites

The samples of farmedfish were collected fromfish farms on

1. In short, milkfish (Chanos chanos, Forsskål, 1775) and mullet (Mugil cephalus, Linnaeus, 1758) are omnivorousfish species which are tolerant to wide ranges of temperatures and salinity and therefore suitable for culture. They have become important marine aquaculture species in the coast of Tanzania. Milkfish from Unguja Island were obtained from Jozani and Shakani ponds (Fig. 1). Jozani ponds are located near Jozani forest famous for colobus monkeys, while Shakani ponds are situated in quarry areas about 3 km from the airport. Milkfish from Pemba island were collected from ponds in Pujini village in Chake Chake town (Fig. 1). Wild mullet were bought fromfisherman at Wete (capital of Pemba island). Farmed and wild milkfish from Mtwara were collected from the ponds and nearby ocean in Ndubwe village. Mtwara region in the southern part of Tanzania, is well-known for the production of cashew nuts, oil and gas, and cement (Fig. 1). The study intended to include analyses offish feed from thefish farms for elucidating contami-nant pathways, however, this was not possible.

2.2. Ethical clearance and permissions to conduct research Permission to conduct this research in the selected sites was given by the management of Institute of Marine Sciences, Univer-sity of Dar es Salaam. Local authorities andfish farm owners were informed about the research aims, and sampling was conducted upon farmers' consent. The permission to transport samples from Tanzania to Norway was granted by The Ministry of Agriculture, Livestock and Fisheries and The Norwegian Food Safety Authority.

2.3. Sampling and sample treatment

Sampling was done in Unguja, Pemba and Mtwara in January, March and April 2016, respectively. A total of 121fish were obtained from bothfish farms and the ocean. Thefish were euthanized with a blow to the head. Dissection and collection of tissue samples were conducted on site except for Shakani site. Due to the high tem-perature (up to 40C) and absence of suitable shade at Shakani, live fish were transported in plastic containers to the laboratory at the Institute of Marine Sciences (IMS) for collection of tissue samples.

Samples were immediately put on dry ice or in liquid nitrogen, depending on the following analysis, as described below.

Overview of the analysed samples andfish characteristics is presented in Table 1. Dissection took place on aluminium foil cleaned with ethanol. Liver samples were put in 10 mL labelled plastic vials; muscles were wrapped in labelled aluminium foil and put in a zipper bag, then immediately transferred into the cool box with ice. To avoid cross contamination, tools were cleaned with ethanol between eachfish sampling. Samples were transferred to IMS and kept in a freezer at20^OC. For sites that required over a day of transportation, samples were stored in a freezer until the traveling time; then transported in a cool box with ice to IMS and then stored in the freezer at20^OC until transportation to Norway for analysis. During transportation to Norway the samples were kept frozen, and after the arrival to Norway the samples were stored in a freezer at20^OC until analysis.

2.4. Sample analysis

At each location thefish was selected on weight and length and grouped (Table 1). The intention was to get a sufficient number of fish with the same weight per site and per species, however, at some sites the availablefish were smaller, as in Mtwara. Chemical

E.B. Mwakalapa et al. / Chemosphere 191 (2018) 438e449 439

the requirement of the NS-EN ISO/IEC 17025 (TEST 137). Thefish liver samples were analysed for OCPs, PCBs and BFRs. Due to small sample seize of liver tissue, perfluoroalkyl substances (PFAS) were analysed infish muscles.

available. One sample was lost during sample preparation, leaving 47fish liver samples for analyses for OCPs: hexachlorobenzene (HCB), a, b- and g-hexachlorocyclohexanes (P

HCHs), oxy-chlordane, trans-chlordane, cis-chlordane and trans-nonachlor (PCHLs), mirex, bis-2,2-(4-chlorophenyl)-1,1,1- trichloroethane

0 0 0 0

Fig. 1.A map of Tanzanian coasts showing the location of sampling sites.

Table 1

Sites andfish characteristics: Sampling time, salinity, mean and range of individual weight and length and number of individual samples of liver from farmed milkfish from Jozani and Shakani (Unguja), Mtwara and Pemba, wild milkfish from Mtwara and wild mullets from Pemba, Tanzania.

Site Fish type Sampling time Salinity (ppt) Mean weight (g) Weight range (g) Mean length (cm) No of sample

Jozani ponds Milkfish Jan-16 36 662 413e826 44.0625 8

Shakani ponds Milkfish Jan-16 40 683.375 533e936 43.5 8

Pemba Ponds Milkfish Mar-16 25 211.875 196e226 29.4375 8

Pemba wild Mullets Mar-16 30 611.75 542e711 39.6357 8

Mtwara ponds Milkfish Apr-16 22 189.05 83.8e308.6 25.8125 8

Mtwara wild Milkfish Apr-16 29 107.55 59.2e185.2 21.957 7

E.B. Mwakalapa et al. / Chemosphere 191 (2018) 438e449 440

BDE-28, -47,99,100, 153,154,183,206,207,208 and209 (P

11PBDEs) and hexabromocyclododecane (HBCDD).

Before weighing,fish liver samples were macerated with scalpel and thoroughly mixed to obtain homogeneous samples. Approxi-mately 0.5e1 g of homogenizedfish liver was weighed into pre-cleaned glass centrifuge tubes. After weighing, 25mL of internal standards PCB -29,112 and207 (1000mg/mL) (Ultra-Scientific, RI, USA); 20mL of BDE -77,119,181, and¹³C12-209,¹³C12-TBBP-A (500mg/mL) (Cambridge Isotope Laboratories, Inc., MA, USA) were added in all the samples. Furthermore, 10 mL distilled water, 2 mL 6% sodium chloride (NaCl), 15 mL acetone and 20 mL cyclohexane were added. The mixture of sample tissue, standards andfluids were homogenized using Ultra Turax homogenizer (IKA Ultra-Turrax T25, IKA Laboratory Technology, Staufen, Germany).

Thereafter followed by lipid extraction using ultrasonic homoge-nizer (Cole Parmer CPX 750, Vernon Hills IL, USA) The second extraction was done with 5 mL acetone and 10 mL cyclohexane.

Separation of the lipid extract from the sample was done by centrifuging at 2095g for 10 min using Allegra X-12R Centrifuge (Beckman Coulter, Fullerton, CA, USA). The method for analysis used is based onBrevik (1978)with modification as described in Polder et al. (2008b). The lipid extract was concentrated by evap-oration using the Zymark Turbo Vap II evaporator (Zymark Coop-eration, Hopkinton, MA, USA) at 40 C and thereafter lipid determination was done gravimetrically using 1 mL aliquot of fat extract. The clean-up of the remaining lipid extract was performed using 96% H2SO4(Fluka Analytika, Sigma-Aldrich, St. Louis, USA).

Thefinal extracts were evaporated on a sand bath at 40C with the blow of N2and concentrated to afinal volume of 0.5 mL before transferred into the 2 mL amber vials for GC analysis.

2.4.1.2. PFAS.Due to lack of liver tissue after analyses of OCPs, PCBs and BFRs, muscle was used as matrix for analysing PFASs. Out of 121 fishes, muscle of the same 47fish were analysed for 2 PFSAs groups:

perfluorohexane sulfonate (PFHxS) and perfluorooctane sulfonate (PFOS) and 6 PFCAs: perfluorooctanoic acid (PFOA), per-fluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), per-fluoroundecanoic acid (PFUdA), perfluorododecanoic acid (PFDoA), and perfluorotridecanoic acid (PFTrDA), summarized asS8PFAS.

The muscle samples werefinely chopped with a scalpel before weighing. Approximately 1 g was weighed into the plastic centri-fuge tubes. 40mL internal standards I.S PFAS Syrene (500 ng/mL) and 40mL I.S PFAS Sulfonate (500 ng/mL) (Wellington Laboratories Inc, CANADA) were added in all the samples followed by the addition of 5 mL of methanol. After adding methanol, the mixture was homogenized using Ultra Turax T25 Homogenizer then further sonicated using Ultra Sonic processor. Extraction was done twice with methanol by shaking the sonicated samples in Vibrax VXR (IKA Werke GmbH&Co. KG, Germany) machine for 30 min fol-lowed by centrifugation at 2095g for 10 min. The extracts were concentrated by evaporation in a Turbo Vap LV evaporator at 40C before cleaning up the lipids with active carbon (Env Carb). After lipid clean-up, the extracts were concentrated again by evaporation to afinal volume of 1 mL and then transferred into the LC-MS vials.

During the analytical procedure sample extracts were not in con-tact with any glassware to avoid the possible adsorption of the analytes.

2.4.2. Instrumental analysis

2.4.2.1. Separation and detection of the POPs.OCPs, PCBs and BFRs were separated and detected using GC-MS methods, PFASs were

selected ion monitoring (SIM), see list with target ions in Suppl S 1.

3.

2.4.2.2. OCPs and PCBs. Separation and detection of chlorinated compounds were performed on a HRGC (Agilent 6890 Series) coupled to a MS detector (Agilent 5975C Agilent Technologies) which was operated in negative chemical ionization (NCI) mode with selected ion monitoring (SIM) as described inPolder et al.

(2014). The OC compounds were separated on a DB-5 MS column

(J&W Scientific, Agilent Technologies) (60 m, 0.25 mm i.d., 0.25mm

film thickness). The temperature program was: 90C (3 min hold);

25C/min increase to 180C (2 min hold); 1.5C/min increase to 220C (2 min hold); and 3C/min increase to 275C (15 min hold) and 25C/min increase to 300C (1 min hold). The total run time was 70.6 min. The carrier gas was hydrogen (H2) at a 1.3 mL/min constantflow. The injection volume was 2mL.

2.4.2.3. BFRs.Detection of PBDEs (except from BDE-209) and HBCDD was performed on a HRGCeLRMS (Agilent 6890 Series;

Agilent Technologies), equipped with an auto-sampler (Agilent 7683 Series; Agilent Technologies) and coupled to a MS detector (Agilent 5973 Network; Agilent Technologies) (Polder et al., 2014).

In short, the separation and identification of the compounds were performed on a DB-5 MS column (30 m, 0.25 mm i.d., 0.25mmfilm thickness; J&W Scientific). The injection volume was 2mL. For detection of BDE-209, extracts (10mL) were injected on a GCeMS (Agilent 6890 Series/5973Network) configured with a program-mable temperature vaporization (PTV) injector (Agilent Technolo-gies). The separation and identification of BDE-209 were performed on a DB-5-MS column (10 m, 0.25 mm i.d., 0.10mmfilm thickness;

J&W Scientific, Agilent Technologies).

2.4.2.4. PFASs.Separation and detection of PFASs were conducted using the LC-MS/MS (API 3000; Applied bioscience, MDX SCIEX) composing of API 3000 triple-quadrupole mass spectrometer coupled to Agilent 1100 HPLC (Agilent Technologies), as described byGrønnestad et al. (2017). The injection volume was 10mL.In short, the column characteristics include 2.1 mm inner diameter and 150 mm long, with a particle size of 3.5mm (Agilent Eclipse Plus C18). The operation software was the Analyst version 1.6.

2.5. QA/QC

2.5.1. OCPs, PCBs, BFRs

Every analytical series included one blind sample of non-spiked salmon trout (Salmo trutta), two samples of spiked salmon trout for recovery, three procedural blanks of solvents and the laboratory's own reference material of the blubber of a harp seal (Pagophilus groenlandicus).The analytical quality was successfully approved by routinely analysing different Certified Reference Materials (CRMs).

In addition, the laboratory successfully participated in Arctic Monitoring and Assessment Program (AMAP) ring test for PCBs, OCPs and PBDEs in human serum 2016, and Quasimeme 2016, round 1: QOR126BT, QOR127BT, QBC046BT, QBC047BT for OCs in fish muscle,fish liver and shellfish tissue inter-laboratory studies.

The limits of detections (LOD) for individual analytes were defined as 3 times the noise level of each analyte. The LODs (ng/g ww) ranged from 0.001 to 0.061 for OCPs, 0.001 to 0.117 for PCBs and 0.001 to 0.196 for BFRs. The relative recoveries for OCPs were be-tween 84 and 118%, for PCBs bebe-tween 81 and 117% and for BFRs between 87 and 112%. The results above and below the limit

E.B. Mwakalapa et al. / Chemosphere 191 (2018) 438e449 441

Atlantic cod (Gadus morhua), two recoveries of spiked Atlantic cod and three blanks of solvent. The analytical quality of the method was assessed by including an inter-laboratory test (AMAP) in the analysis of samples. The LOD for PFAS ranged between 0.08 ng/g ww to 0.43 ng/g ww and the recoveries were between 97 and 125%.

2.5.3. Statistical data analysis

Data were organised in spread sheets (MS Excel, 2016). JMP 11 statistical software was used for further analysis. Compounds detected in less than 60% of the samples were only reported in range and not included in statistical analysis. The compounds which were detected in more than 60% of the samples were re-ported in mean, median and range and were included in further statistical analysis. The non-detects (nd) were treated as zero dur-ing analysis as the concentrations of POPs in general were very low.

Shapiro-Wilk Test W was used to test for the distribution of the data. Since the data were not normally distributed even after transformation, a non-parametric Kruskal-Wallis test was used to test for differences among sites and Tukey test was used to identify the means that were different from each other. Spearman rank correlation was used to assess the correlation between variables.

The difference between sites were considered statistically signifi-cant whenp<0.05.

3. Results

3.1. Site andfish characteristics

Salinity varied among locations and is presented inTable 1. The mean weights of farmed milkfish from Jozani and Shakani were higher than the farmed and wild milkfish from Mtwara and farmed milkfish from Pemba (Table 1). The mean lipid percentage infish liver ranged between 3.1% in farmed milkfish from Pemba and 11.4%

in farmed milkfish from Shakani (Table 2).

3.2. Levels of OCPs

DDTs, HCB andg-HCH were detected in more than 90% of the samples (Suppl S 3.2). DDTs were the dominating OCPs in milkfish and mullet with the highest mean concentrations in all sites (Table 2). The mean percent contributions to theP

DDTs were 84.4% forp,p⁰-DDE, 8.4% forp,p⁰-DDD, 7.1% forp,p⁰-DDT, and 0.2% for o,p⁰-DDT (Suppl S 3.1).p,p⁰-DDE was the dominating compound in all sites with highest maximum concentrations of 715 ng/g lw in wild milkfish from Mtwara (Suppl S 2.1). The ratios ofp,p⁰-DDE/p,p⁰ -DDT (Suppl S 2.1) ranged from 3.4 to 6.6 (mean 4.6) in farmed milkfish from Jozani, 6.4e9.3 (mean 7.6) in farmed milkfish from Shakani, 5.3e10.2 (mean 7.1) in farmed milkfish from Mtwara, 40.9 (only one ratio available) in wild milkfish from Mtwara and 15.1e95.9 (mean 34.6) in wild mullet from Pemba. No ratio was available for farmed milkfish from Pemba becausep,p⁰-DDE was the only DDT metabolite detected. The highest concentration of HCB was 0.52 ng/g lw found in farmed milkfish from Shakani (Table 2;

Fig. 2).g-HCH contributed 97% toP

HCHs. The highest level ofg -HCH was detected in wild milkfish from Mtwara (0.19 ng/g lw)

HCHs. The highest level ofg -HCH was detected in wild milkfish from Mtwara (0.19 ng/g lw)

In document Persistent Organic Pollutants (POPs) and heavy metals in fish from Tanzania : levels and associated health risks to humans and fish (sider 54-188)