Differences in AChE and BChE Enzyme Activation Levels in Liver and Brain Tissues in Rainbow Trouts Exposed to Different Bacterial Diseases

Abstract

In this study, symptomatic fish samples were taken from rainbow trout farms. Isolation and identification of agents isolated from fish samples were made. DNA isolations from different purified colonies were carried out with the mericon bacterial DNA kit. Real-Time PCR procedure was performed by using universal bacterial primers. Molecular identifications were performed by blasting the nucleotides obtained by sequence analysis of PCR amplicons. Spectrophotometric measurements were performed at 412 nm wavelengths for AChE activity and 412 nm for BChE activity from liver and brain tissues of fish samples. The activity differences of different disease factors among themselves and according to the control group were examined. As a result of the study, isolation and identification of Bacillus subtilis, Lactococcus garvieae and Staphylococcus epidermidis from 5 different farms were performed. Over 98% similarity was observed as a result of sequencing analysis of the isolates. In this study, it was observed that three different bacteria isolated from trout farms suppressed AChE and BChE enzyme activities in both tissues of trout.

Keywords

• AChE
• Bacteria
• BChE
• Rainbow trout
• Real-Time PCR
• Sequencing

Farklı Bakteriyel Hastalıklara Maruz Kalmış Gökkuşağı Alabalıklarında Karaciğer ve Beyin Dokularında AChE ve BChE Enzim Aktivite Seviyesi Farklılıkları

Öz

Bu çalışmada, gökkuşağı alabalığı işlemelerinden semptomlu balık örnekleri alınmıştır. Balık örneklerinden etkenlerinin izolasyonu ve identifikasyonları yapılmıştır. Saflaştırılan farklı kolonilerden DNA izolasyonları mericon bacterial DNA kiti ile gerçekleştirilmiştir. Real-Time PCR prosedürü evrensel bakteriyel primerler kullanılarak gerçekleştirilmiştir. Moleküler identifikasyonlar PCR amplikonlarının dizi analizi ile elde edilen nükleotidlerin blastlaması ile gerçekleştirilmiştir. Balık örneklerinin karaciğer ve beyin dokularından AChE aktivitesi için 412 nm ve BChE aktivitesi için 412 nm dalga boylarında spektrofotometrik ölçümler yapılmıştır. Farklı hastalık etkenlerinin kendi aralarında ve kontrol grubuna göre aktivite farklılıkları incelenmiştir. Çalışma sonucunda, 5 farklı işletmelerin 3’ünden Bacillus subtilis, Lactococcus garvieae ve Staphylococcus epidermidis etkenlerinin izolasyon ve identifikasyonları gerçekleştirilmiştir. İzolatların dizileme analizi sonucunda %98'in üzerinde benzerlik gözlenmiştir. Çalışmada, alabalık çiftliklerinden izole edilen üç farklı bakterinin her iki dokusunda da AChE ve BChE enzim aktivitelerini baskıladığı görülmüştür.

Anahtar Kelimeler

• AChE,
• Bakteri
• BChE
• Rainbow trout
• Real-Time PCR

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