Odorous Compounds from Poultry Manure Induce DNA Damage, Nuclear Changes, and Decrease Cell Membrane Integrity in Chicken Liver Hepatocellular Carcinoma Cells

Abstract

Animal breeding and management of organic wastes pose a serious problem to the health of livestock and workers, as well as the nearby residents. The aim of the present study was to determine the mechanisms of toxicity of selected common odorous compounds from poultry manure, including ammonia, dimethylamine (DMA), trimethylamine (TMA), butyric acid, phenol, and indole. We measured their genotoxic and cytotoxic activity in the model chicken cell line (LMH), in vitro, by comet assay and lactate dehydrogenase assay, respectively. We also made microscopic observations of any morphological changes in these cells by DAPI staining. Four compounds, namely ammonia, DMA, TMA, and butyric acid increased DNA damage in a dose-dependent manner (p < 0.05), reaching genotoxicity as high as 73.2 ± 1.9%. Phenol and indole induced extensive DNA damage independent of the concentration used. Ammonia, DMA, and TMA caused a dose-dependent release of lactate dehydrogenase (p < 0.05). The IC50 values were 0.02%, 0.05%, and 0.1% for DMA, ammonia and TMA, respectively. These compounds also induced nuclear morphological changes, such as chromatin condensation, shrinkage, nuclear fragmentation (apoptotic bodies), and chromatin lysis. Our study exhibited the damaging effects of odorous compounds in chick LMH cell line.

Synchrotron-Based Fourier-Transform Infrared Micro-Spectroscopy (SR-FTIRM) Fingerprint of the Small Anionic Molecule Cobaltabis(dicarbollide) Uptake in Glioma Stem Cells

“…Cytotoxicity and EC 50 were determined by PrestoBlue assays according to Nowak et al and Organization for Economic Cooperation and Development (OECD) protocol [ 78 ]. The absorbance of the control sample (untreated cells) represented 100% cell viability.…”

Acrylamide Decreases Cell Viability, and Provides Oxidative Stress, DNA Damage, and Apoptosis in Human Colon Adenocarcinoma Cell Line Caco-2

“…Exposure to 6.4-50 mM of AA (24 h), 3.2-50 mM of AA (48 h), and 0.8-50 mM of AA (72 h) showed a significant increase in AA cytotoxicity in the MTT assay, while in the PrestoBlue assay, AA induced cytotoxic effects from 6.4-50 mM of AA (24 h) and 1.6-50 mM of AA (48-72 h) (p < 0.05). The inhibitory concentration (IC) 50 values after 24-72 h of exposure to AA showed higher cytotoxicity in the MTT assay (5.9, 2.5, and 0.7 mM) than PrestoBlue assay (8.9, 3.9, and 2.6 mM), respectively. Different values obtained for each of the assay types resulted from the diverse molecular mechanism used by them.…”

“…Cytotoxicity and IC 50 were determined by MTT and PrestoBlue assays according to Nowak et al [50] and Organisation for Economic Cooperation and Development (OECD) protocol [51]. The absorbance/fluorescence of the control sample (untreated cells) represented 100% cell viability.…”

“…The final amount of each sample was set to 1 mL. The comet assay was performed under alkaline conditions (pH > 13) as previously described [50]. After incubation, aliquots of suspended cells were centrifuged (182× g, 15 min, 4 • C); decanted; suspended in 0.75% LMP agarose; distributed onto slides precoated with 0.5% NMP agarose; and immersed in lysing solution consisting of 2.5 M NaCl, 1% Triton X-100, 100 mM EDTA, and 10 mM Tris, at pH 10 (4 • C, 1 h).…”

Influence of Certain Animal Manures on Nematode Community in Mandarin Orchards Citrus reticulata (Blanco) in Sharkia Governorate, Egypt

“…Among which direct effects of chemicals produced during manure decomposition such as ammonia, hydrogen sulphide ,methan, fatty acids with low molecular weight like acetic, propionic acids, dimethlamine , trimethyamine as well as phenols. All these materials were reported to be deleterious to different species of plant-parasitic nematodes ( Badra et al, 1979 ;Main & Rodriguez -Kabana, 1982;Rodriguez-Kabana, 1986;Kesba &Al-Shalaby, 2008 andNowak et al, 2017).…”