Search results for: methanogenic toxicity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 941

Search results for: methanogenic toxicity

941 Effect of Antimony on Microorganisms in Aerobic and Anaerobic Environments

Authors: Barrera C. Monserrat, Sierra-Alvarez Reyes, Pat-Espadas Aurora, Moreno Andrade Ivan

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Antimony is a toxic and carcinogenic metalloid considered a pollutant of priority interest by the United States Environmental Protection Agency. It is present in the environment in two oxidation states: antimonite (Sb (III)) and antimony (Sb (V)). Sb (III) is toxic to several aquatic organisms, but the potential inhibitory effect of Sb species for microorganisms has not been extensively evaluated. The fate and possible toxic impact of antimony on aerobic and anaerobic wastewater treatment systems are unknown. For this reason, the objective of this study was to evaluate the microbial toxicity of Sb (V) and Sb (III) in aerobic and anaerobic environments. Sb(V) and Sb(III) were used as potassium hexahydroxoantimonate (V) and potassium antimony tartrate, respectively (Sigma-Aldrich). The toxic effect of both Sb species in anaerobic environments was evaluated on methanogenic activity and the inhibition of hydrogen production of microorganisms from a wastewater treatment bioreactor. For the methanogenic activity, batch experiments were carried out in 160 mL serological bottles; each bottle contained basal mineral medium (100 mL), inoculum (1.5 g of VSS/L), acetate (2.56 g/L) as substrate, and variable concentrations of Sb (V) or Sb (III). Duplicate bioassays were incubated at 30 ± 2°C on an orbital shaker (105 rpm) in the dark. Methane production was monitored by gas chromatography. The hydrogen production inhibition tests were carried out in glass bottles with a working volume of 0.36 L. Glucose (50 g/L) was used as a substrate, pretreated inoculum (5 g VSS/L), mineral medium and varying concentrations of the two species of antimony. The bottles were kept under stirring and at a temperature of 35°C in an AMPTSII device that recorded hydrogen production. The toxicity of Sb on aerobic microorganisms (from a wastewater activated sludge treatment plant) was tested with a Microtox standardized toxicity test and respirometry. Results showed that Sb (III) is more toxic than Sb (V) for methanogenic microorganisms. Sb (V) caused a 50% decrease in methanogenic activity at 250 mg/L. In contrast, exposure to Sb (III) resulted in a 50% inhibition at a concentration of only 11 mg/L, and an almost complete inhibition (95%) at 25 mg/L. For hydrogen-producing microorganisms, Sb (III) and Sb (V) inhibited 50% of this production with 12.6 mg/L and 87.7 mg/L, respectively. The results for aerobic environments showed that 500 mg/L of Sb (V) do not inhibit the Allivibrio fischeri (Microtox) activity or specific oxygen uptake rate of activated sludge. In the case of Sb (III), this caused a loss of 50% of the respiration of the microorganisms at concentrations below 40 mg/L. The results obtained indicate that the toxicity of the antimony will depend on the speciation of this metalloid and that Sb (III) has a significantly higher inhibitory potential compared to Sb (V). It was shown that anaerobic microorganisms can reduce Sb (V) to Sb (III). Acknowledgments: This work was funded in part by grants from the UA-CONACYT Binational Consortium for the Regional Scientific Development and Innovation (CAZMEX), the National Institute of Health (NIH ES- 04940), and PAPIIT-DGAPA-UNAM (IN105220).

Keywords: aerobic inhibition, antimony reduction, hydrogen inhibition, methanogenic toxicity

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940 Pre-Treatment of Anodic Inoculum with Nitroethane to Improve Performance of a Microbial Fuel Cell

Authors: Rajesh P.P., Md. Tabish Noori, Makarand M. Ghangrekar

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Methanogenic substrate loss is reported to be a major bottleneck in microbial fuel cell which significantly reduces the power production capacity and coulombic efficiency (CE) of microbial fuel cell (MFC). Nitroethane is found to be a potent inhibitor of hydrogenotrophic methanogens in rumen fermentation process. Influence of nitroethane pre-treated sewage sludge inoculum on suppressing the methanogenic activity and enhancing the electrogenesis in MFC was evaluated. MFC inoculated with nitroethane pre-treated anodic inoculum demonstrated a maximum operating voltage of 541 mV, with coulombic efficiency and sustainable volumetric power density of 39.85 % and 14.63 W/m3 respectively. Linear sweep voltammetry indicated a higher electron discharge on the anode surface due to enhancement of electrogenic activity while suppressing methanogenic activity. A 63 % reduction in specific methanogenic activity was observed in anaerobic sludge pre-treated with nitroethane; emphasizing significance of this pretreatment for suppressing methanogenesis and its utility for enhancing electricity generation in MFC.

Keywords: coulombic efficiency, methanogenesis inhibition, microbial fuel cell, nitroethane

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939 Application of Customized Bioaugmentation Inocula to Alleviate Ammonia Toxicity in CSTR Anaerobic Digesters

Authors: Yixin Yan, Miao Yan, Irini Angelidaki, Ioannis Fotidis

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Ammonia, which derives from the degradation of urea and protein-substrates, is the major toxicant of the commercial anaerobic digestion reactors causing loses of up to 1/3 of their practical biogas production, which reflects directly on the overall revenue of the plants. The current experimental work is aiming to alleviate the ammonia inhibition in anaerobic digestion (AD) process by developing an innovative bioaugmentation method of ammonia tolerant methanogenic consortia. The ammonia tolerant consortia were cultured in batch reactors and immobilized together with biochar in agar (customized inocula). Three continuous stirred-tank reactors (CSTR), fed with the organic fraction of municipal solid waste at a hydraulic retention time of 15 days and operated at thermophilic (55°C) conditions were assessed. After an ammonia shock of 4 g NH4+-N L-1, the customized inocula were bioaugmented into the CSTR reactors to alleviate ammonia toxicity effect on AD process. Recovery rate of methane production and methanogenic activity will be assessed to evaluate the bioaugmentation performance, while 16s rRNA gene sequence will be used to reveal the difference of microbial community changes through bioaugmentation. At the microbial level, the microbial community structures of the four reactors will be analysed to find the mechanism of bioaugmentation. Changes in hydrogen formation potential will be used to predict direct interspecies electron transfer (DIET) between ammonia tolerant methanogens and syntrophic bacteria. This experimental work is expected to create bioaugmentation inocula that will be easy to obtain, transport, handled and bioaugment in AD reactors to efficiently alleviate the ammonia toxicity, without alternating any of the other operational parameters including the ammonia-rich feedstocks.

Keywords: artisanal fishing waste, acidogenesis, volatile fatty acids, pH, inoculum/substrate ratio

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938 Gamma-Hydroxybutyrate (GHB): A Review for the Prehospital Clinician

Authors: Theo Welch

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Background: Gamma-hydroxybutyrate (GHB) is a depressant of the central nervous system with euphoric effects. It is being increasingly used recreationally in the United Kingdom (UK) despite associated morbidity and mortality. Due to the lack of evidence, healthcare professionals remain unsure as to the optimum management of GHB acute toxicity. Methods: A literature review was undertaken of its pharmacology and the emergency management of its acute toxicity.Findings: GHB is inexpensive and readily available over the Internet. Treatment of GHB acute toxicity is supportive. Clinicians should pay particular attention to the airway as emesis is common. Intubation is required in a minority of cases. Polydrug use is common and worsens prognosis. Conclusion: An inexpensive and readily available drug, GHB acute toxicity can be difficult to identify and treat. GHB acute toxicity is generally treated conservatively. Further research is needed to ascertain the indications, benefits, and risks of intubating patients with GHB acute toxicity. instructions give you guidelines for preparing papers for the conference.

Keywords: GHB, gamma-hydroxybutyrate, prehospital, emergency, toxicity, management

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937 Performance of an Anaerobic Baffled Reactor (ABR) during Start-Up Period

Authors: D. M. Bassuney, W. A. Ibrahim, Medhat A. E. Moustafa

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Appropriate start-up of an anaerobic baffled reactor (ABR) is considered to be the most delicate and important issue in the anaerobic process, and depends on several factors such as wastewater composition, reactor configuration, inoculum and operating conditions. In this work, the start-up performance of an ABR with working volume of 30 liters, fed continuously with synthetic food industrial wastewater along with semi-batch study to measure the methangenic activity by specific methanogenic activity (SMA) test were carried out at various organic loading rates (OLRs) to determine the best OLR used to start up the reactor. The comparison was based on COD removal efficiencies, start-up time, pH stability and methane production. An OLR of 1.8 Kg COD/m3d (5400 gCOD/m3 and 3 days HRT) showed best overall performance with COD removal efficiency of 94.44% after four days from the feeding and methane production of 3802 ml/L with an overall SMA of 0.36 gCH4-COD/gVS.d

Keywords: anaerobic baffled reactor, anaerobic reactor start-up, food industrial wastewater, specific methanogenic activity

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936 The Effects of Nano Zerovalent Iron (nZVI) and Magnesium Oxide Nanoparticles on Methane Production during Anaerobic Digestion of Waste Activated Sludge

Authors: Passkorn Khanthongthip, John T. Novak

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Many studies have been reported that the nZVI and MgO NPs were often found in waste activated sludge (WAS). However, little is known about the impact of those NPs on WAS stabilization. The aims of this study were to investigate the effects of both NPs on WAS anaerobic digestion for methane production and to examine the change of metanogenic population under those different environments using qPCR. Four dosages (2, 50, 100, and 200 mg/g-TSS) of MgO NPs were added to four different bottles containing WAS to investigate the impact of MgO NPs on methane production during WAS anaerobic digestion. The effects of nZVI on methane production during WAS anaerobic digestion were also conducted in another four bottles using the same methods described above except that the MgO NPs were replaced by nZVI. A bottle of WAS anaerobic digestion without nanoparticles addition was also operated to serve as a control. It was found that the relative amounts, compared to the control system, of methane production in each WAS anaerobic digestion bottle adding 2, 50, 100, 200 mg/gTSS MgO NPs were 98, 62, 28, and 14 %, respectively. This suggests that higher MgO NPs resulted in lower methane production. The data of batch test for the effects of corresponding released Mg2+ indicated that 50 mg/gTSS MgO NPs or higher could inhibit methane production at least 25%. Moreover, the volatile fatty acid (VFA) concentration was 328, 384, 928, 3,684, and 7,848 mg/L for the control and four WAS anaerobic digestion bottles with 2, 50, 100, 200 mg/gTSS MgO NPs addition, respectively. Higher VFA concentration could reduce pH and subsequently decrease methanogen growth, resulting in lower methane production. The relative numbers of total gene copies of methanogens analyzed from samples taken from WAS anaerobic digestion bottles were approximately 99, 68, 38, and 24 % of control for the addition of 2, 50, 100, and 200 mg/gTSS, respectively. Obviously, the more MgO NPs appeared in sludge anaerobic digestion system, the less methanogens remained. In contrast, the relative amount of methane production found in another four WAS anaerobic digestion bottles adding 2, 50, 100, and 200 mg/gTSS nZVI were 102, 128, 112, and 104 % of the control, respectively. The measurement of methanogenic population indicated that the relative content of methanogen gene copies were 101, 132, 120, and 112 % of those found in control, respectively. Additionally, the cumulative VFA was 320, 234, 308, and 330 mg/L, respectively. This reveals that nZVI addition could assist to increase methanogenic population. Higher amount of methanogen accelerated VFA degradation for greater methane production, resulting in lower VFA accumulation in digesters. Moreover, the data for effects of corresponding released Fe2+ conducted by batch tests suggest that the addition of approximately 50 mg/gTSS nZVI increased methane production by 20%. In conclusion, the presence of MgO NPs appeared to diminish the methane production during WAS anaerobic digestion. Higher MgO NPs dosages resulted in more inhibition on methane production. In contrast, nZVI addition promoted the amount of methanogenic population which facilitated methane production.

Keywords: magnesium oxide nanoparticles, methane production, methanogenic population, nano zerovalent iron

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935 Estimation of the Acute Toxicity of Halogenated Phenols Using Quantum Chemistry Descriptors

Authors: Khadidja Bellifa, Sidi Mohamed Mekelleche

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Phenols and especially halogenated phenols represent a substantial part of the chemicals produced worldwide and are known as aquatic pollutants. Quantitative structure–toxicity relationship (QSTR) models are useful for understanding how chemical structure relates to the toxicity of chemicals. In the present study, the acute toxicities of 45 halogenated phenols to Tetrahymena Pyriformis are estimated using no cost semi-empirical quantum chemistry methods. QSTR models were established using the multiple linear regression technique and the predictive ability of the models was evaluated by the internal cross-validation, the Y-randomization and the external validation. Their structural chemical domain has been defined by the leverage approach. The results show that the best model is obtained with the AM1 method (R²= 0.91, R²CV= 0.90, SD= 0.20 for the training set and R²= 0.96, SD= 0.11 for the test set). Moreover, all the Tropsha’ criteria for a predictive QSTR model are verified.

Keywords: halogenated phenols, toxicity mechanism, hydrophobicity, electrophilicity index, quantitative stucture-toxicity relationships

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934 Metal Nanoparticles Caused Death of Metastatic MDA-MB-231 Cells

Authors: O. S. Adeyemi, C. G. Whiteley

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The present study determined the toxic potential of metal nanoparticles in cell culture system. Silver and gold nanoparticles were synthesized and characterized following established "green" protocols. The synthesized nanoparticles, in varying concentrations ranging from 0.1–100 µM were evaluated for toxicity in metastatic MDA-MB-231 cells. The nanoparticles promoted a generation of reactive oxygen species and reduced cell viability to less than 50% in the demonstration of cellular toxicity. The nanoparticles; gold and the silver-gold mixture had IC50 values of 56.65 and 18.44 µM respectively. The IC50 concentration for silver nanoparticles could not be determined. Furthermore, the probe of the cell death using flow cytometry and confocal microscopy revealed the partial involvement of apoptosis as well as necrosis. Our results revealed cellular toxicity caused by the nanoparticles but the mechanism remains yet undefined.

Keywords: cell death, nanomedicine, nanotoxicology, toxicity

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933 Toxicological Study of Umbilicus rupesris L. Leaves: Hematological, Biochemical, and Histopathological Studies

Authors: Afaf Benhouda, Mouloud Yahia, Hachani Khadraoui, Asma Meddour, Souhila Benbia, Abdelmoudjib Ghecham, Djahida Benhouda

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Umbilicus rupestris (UR) is an herbal medicine traditionally applied against the ignitions of the skin. The present paper aimed to study the acute and subacute toxicity with orally administered methanolic leaves extract of Umbilicus rupestris L (URMeOH). In acute toxicity tests, four groups of rats (n = 6/group/female) were orally treated with doses of 500, 1000, 1500 and 2000 mg/kg, and general behaviour, adverse effects, and mortality were recorded for up to 14 days. In subacute toxicity study, rats received URAMeOH by gavage at the doses of 100, 200 mg/kg/day (n = 6/group) for 28 days, and biochemical, hematological, and histopathological changes in tissues (liver, kidney) were determined. URMeOH did not produce any hazardous symptoms or death and in the acute toxicity test. Subacute treatment with URMeOH did not show any change in body weight, and hematological and biochemical profiles. In addition, no change was observed either in macroscopic or microscopic aspects of vital organs in rats. Our result showed that Umbilicus rupestris extract could be safe for human use.

Keywords: acute toxicity, biochemical parameters, hematological parameters, Umbilicus rupestris, subacute toxicity

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932 Antioxidant and Acute Toxicity of Stem Extracts of the Ficus Iteophylla

Authors: Muhammad Mukhtar

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The aim of this study is to evaluate the antioxidant activity and acute toxicity of the extracts of Ficus iteophylla by reactions with 1, 1-diphenyl-2-picryhydrazyl radical (DPPH) and method developed by Lork 1983, respectively. Stem bark of Ficus iteophylla was collected, air dried, pulverized to fine powdered and sequentially extracted using acetone, methanol and water in order of increasing polarity. The result shows strong radical scavenging activity against DPPH for all the extracts when compared with ascorbic acid. The LD50 of 316 mg/kg was calculated for all the three extras, and the values were found to be within the practically toxic range, and therefore, care should be taken when using the plants in traditional medicine.

Keywords: antioxidant, acute toxicity, Ficus iteophylla

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931 Acute Oral Toxicity Study of Mystroxylon aethiopicum Root Bark Aqueous Extract in Albino Mice

Authors: Mhuji Kilonzo

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Acute oral toxicity of Mystroxylon aethiopicum root bark aqueous was evaluated in albino mice of either sex. In this study, five groups of mice were orally treated with doses of 1000, 2000, 3000, 4000 and 5000 mg/kg body weight of the crude extract. The mortality, signs of toxicity and body weights were observed individually for two weeks. At the end of the two weeks study, all animals were sacrificed, and the hematological and biochemical parameters, as well as organ weights relative to body weight of each animal, were determined. No mortality, signs of toxicity and abnormalities in vital organs were observed in the entire period of study for both treated and control groups of mice. Additionally, there were no significant changes (p > 0.05) in the blood hematology and biochemical analysis. However, the body weights of all mice increased significantly. The Mystroxylon aethiopicum root bark aqueous extract were found to have a high safe margin when administered orally. Hence, the extract can be utilized for pharmaceutical formulations.

Keywords: acute oral toxicity, albino mice, Mystroxylon aethiopicum, safety

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930 Determination of Acid Volatile Sulfides–Simultaneously Extracted Metal Relationship and Toxicity in Contaminated Sediment Layer in Mid-Black Sea Coasts

Authors: Arife Simsek, Gulfem Bakan

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Sediment refers to the accumulation of varying amounts of sediment material in natural waters and the formation of bottom sludge. Sediments are the most important sources of pollutants as well as important future sources and carriers of pollutants. The accumulation of pollutants in sediments can cause serious environmental problems for the surrounding areas. Heavy metals (such as Cr, Cd, Al, Pb, Cu, Al, Zn) disrupt the water quality, affect the useful use of sediment, affect the ecosystem and have a toxic effect on the life of the sediment layer. This effect, which accumulates in the aquatic organisms, can enter the human body with the food chain and affect health seriously. Potential metal toxicity can be determined by comparing acid volatile sulfides (AVS) – simultaneously extracted metal (SEM) ratio in anoxic sediments to determine the effect of metals. Determination of the concentration of SEM and AVS is useful in screening sediments for potential toxicity due to the high metal concentration. In the case of SEM/AVS < 0 (anoxic sediment); in terms of AVS biomass production, its toxicity can be controlled. No toxic effects may be observed when SEM / AVS < 0. SEM / AVS > 0 (in the case of oxic sediment); metals with sensitive fraction such as Cu, As, Ag, Zn are stored. In this study, AVS and SEM measurements of sediment samples collected from five different points in the district of Tekkeköy in Samsun province were performed. The SEM - AVS ratio was greater than 0 in all samples. Therefore, it is necessary to test the toxicity against the risks that may occur in the ecosystem.

Keywords: AVS-SEM, Black Sea, heavy metal, sediment, toxicity

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929 DNA Methylation 6mA and Histone Methylation Involved in Multi-/Trans-Generational Reproductive Effects in Caenorhabditis elegans Induced by Atrazine

Authors: Jiechen Yin, Xiang Hong, Ran Liu

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Atrazine (ATR), a widely used triazine herbicide, is an environmental endocrine disruptor that can cause health problems. However, whether there are multi/trans-generational reproductive impacts of ATR have not been studied to our best knowledge. Therefore, in this study, Caenorhabditis elegans was used as a preferable model organism to identify the multi/trans-generational reproductive toxicity of ATR. L1 larvae were exposed to different concentrations (0.0004–40 mg/L) of ATR for 48 h. Successive generations (F1 to F5) were fed without ATR and consecutive exposure. The results showed that ATR exposure during P0 decreased fecundity, including a reduction in fertilized eggs, oocytes, and ovulation rate, delayed gonadal development, and decreased the relative area of the gonad arm and germ cell number. Furthermore, continuous ATR exposure (P0–F5) causes a significant increase in reproductive toxicity in subsequent generations, although no significant toxicity occurred in the P0 generation after exposure to environmental-related concentrations, suggesting that ATR exposure might have cumulative effects. Likewise, parental exposure to ATR caused transgenerational toxicity impairments. Interestingly, reproductive toxicity not development toxicity was transmitted to several generations (F1–F4), and the F2 generation showed the most notable changes. QRT-PCR results showed that genes related to DNA methylation 6mA (damt-1, nmad-1) and histone H3 methylation (mes-4, met-2, set-25, set-2, and utx-1) can also be passed on to offspring. The function of H3K4 and H3K9 methylation were explored by using loss-of-function mutants for set-2, set-25, and met-2. Transmissible reproductive toxicity was absent in met-2(n4256), set-2(ok952), and set-25(n5021) mutants, which suggests that the histone methyltransferases H3K4 and H3K9 activity are indispensable for the transgenerational effect of ATR. Finally, the downstream genes of DNA methylation and histone H3 methylation were determined. ATR upregulated the expression of ZC317.7, hsp-6, and hsp-60. Mitochondrial stress in parental generation dependent transcription 6mA modifiers may establish these epigenetic marks in progeny.

Keywords: ATR, Caenorhabditis elegans, multi-/trans-generation, reproductive toxicity

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928 Sulfur-Containing Diet Shift Hydrogen Metabolism and Reduce Methane Emission and Modulated Gut Microbiome in Goats

Authors: Tsegay Teklebrhan Gebremariam, Zhiliang, Arjan Jonker

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The study investigated that using corn gluten (CG) instead of cornmeal (CM) increased dietary sulfur shifted H₂ metabolism from methanogenesis to alternative sink and modulated microbiome in the rumen as well as hindgut segments of goats. Ruminal fermentation, CH₄ emissions and microbial abundance in goats (n = 24). The experiment was performed using a randomized block design with two dietary treatments (CM and CG with 400 g/kg DM each). Goats in CG increased sulfur, NDF and CP intake and decreased starch intake as compared with those in CM. Goats that received CG diet had decreased dissolved hydrogen (dH₂) (P = 0.01) and dissolved methane yield and emission (dCH₄) (P = 0.001), while increased dH₂S both in the rumen and hindgut segments than those fed CM. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the rumen and hindgut than those fed CM. Higher (P < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. The study suggested that goats fed corn gluten improved the gene copies of microbiota and fibrolytic bacterial species while reducing starch utilizing species in the rumen and hindgut segments as compared with that fed cornmeal. Goats consuming corn gluten had a more enriched methanogenic diversity and reduced Methanobrevibacter, a contributor to CH₄ emissions, as compared with goats fed CM. Corn gluten could be used as an alternative feed to decrease the enteric CH₄ emission in ruminant production.

Keywords: dissolved gasses, methanogenesis, microbial community, metagenomics

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927 Protective Effect of Protocatechuic Acid Alone and in Combination with Ascorbic Acid in Aniline Hydrochloride Induced Spleen Toxicity in Rats

Authors: Aman Upaganlawar, Upasana Khairnar, Chandrashekhar Upasani

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The present study was designed to evaluate the protective effects of protocatechuic acid alone and in combination with ascorbic acid in aniline hydrochloride-induced spleen toxicity in rats. Male Wistar rats of either sex (200-250g) were used and divided into different groups. Spleen toxicity was induced by aniline hydrochloride (100 ppm) in drinking water for 28 days. Treatment group received protocatechuic acid (40 mg/kg/day, p.o), ascorbic acid (40 mg/kg/day, p.o), and combination of protocatechuic acid (20 mg/kg/day, p.o) and ascorbic acid (20 mg/kg/day, p.o) followed by aniline hydrochloride. At the end of treatment period, serum and tissue parameters were evaluated. Rats supplemented with aniline hydrochloride showed a significant alteration in body weight, spleen weight, feed consumption, water intake, hematological parameters (Hemoglobin content, Red Blood Cells, White Blood Cells and Total iron content), tissue parameters (Lipid peroxidation, Reduced glutathione, Nitric oxide content) compared to control group. Histopathology of aniline hydrochloride-induced spleen showed significant damage compared to control rats. Treatment with Protocatechuic acid along with ascorbic acid showed better protection as compared to protocatechuic acid or ascorbic acid alone in aniline hydrochloride-induced spleen toxicity. In conclusion Treatment with protocatechuic acid and ascorbic acid in combination showed significant protection in aniline hydrochloride-induced splenic toxicity in rats.

Keywords: aniline, spleen toxicity, protocatechuic acid, ascorbic acid, antioxidants

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926 Mitigating Ruminal Methanogenesis Through Genomic and Transcriptomic Approaches

Authors: Muhammad Adeel Arshad, Faiz-Ul Hassan, Yanfen Cheng

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According to FAO, enteric methane (CH4) production is about 44% of all greenhouse gas emissions from the livestock sector. Ruminants produce CH4 as a result of fermentation of feed in the rumen especially from roughages which yield more CH4 per unit of biomass ingested as compared to concentrates. Efficient ruminal fermentation is not possible without abating CO2 and CH4. Methane abatement strategies are required to curb the predicted rise in emissions associated with greater ruminant production in future to meet ever increasing animal protein requirements. Ecology of ruminal methanogenesis and avenues for its mitigation can be identified through various genomic and transcriptomic techniques. Programs such as Hungate1000 and the Global Rumen Census have been launched to enhance our understanding about global ruminal microbial communities. Through Hungate1000 project, a comprehensive reference set of rumen microbial genome sequences has been developed from cultivated rumen bacteria and methanogenic archaea along with representative rumen anaerobic fungi and ciliate protozoa cultures. But still many species of rumen microbes are underrepresented especially uncultivable microbes. Lack of sequence information specific to the rumen's microbial community has inhibited efforts to use genomic data to identify specific set of species and their target genes involved in methanogenesis. Metagenomic and metatranscriptomic study of entire microbial rumen populations offer new perspectives to understand interaction of methanogens with other rumen microbes and their potential association with total gas and methane production. Deep understanding of methanogenic pathway will help to devise potentially effective strategies to abate methane production while increasing feed efficiency in ruminants.

Keywords: Genome sequences, Hungate1000, methanogens, ruminal fermentation

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925 Toxicity Identification and Evaluation for the Effluent from Seawater Desalination Facility in Korea Using D. magna and V. fischeri

Authors: Sung Jong Lee, Hong Joo Ha, Chun Sang Hong

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In recent years, the interests on the impacts of industrial wastewater on aquatic ecosystem have increased with concern about ecosystem protection and human health. Whole effluent toxicity tests are used to monitor toxicity by unknown toxic chemicals as well as conventional pollutants from industrial effluent discharges. This study describes the application of TIE (toxicity identification evaluation) procedures to an acutely toxic effluent from a Seawater desalination facility in industrial complex which was toxic to Daphnia magna. In TIE phase I (characterization step), the toxic effects by heavy metals, organic compounds, oxidants, volatile organic compounds, suspended solids and ammonia were screened and revealed that the source of toxicity is far from these toxicants group. Chemical analysis (TIE phase II) on TDS showed that the concentration of chloride ion (24,215 ~ 29,562 mg/L) was substantially higher than that predicted from EC50 for D. magna. In confirmation step (TIE phase III), chloride ion was demonstrated to be main toxicant in this effluent by the spiking approach, species sensitivity approach, and deletion approach. Calcium, potassium, magnesium, sodium, fluorine, sulfate ion concentration was not shown toxicity from D. magna. Finally, we concluded that chloride was the most contributing toxicant in the waste water treatment plant. Further research activities are needed for technical support of toxicity identification and evaluation on the various types of wastewater treatment plant discharge in Korea. Acknowledgement: This research was supported by a grant (16IFIP-B089911-03) from Plant Research Program funded by Ministry of Land, Infrastructure and Transport of Korean government.

Keywords: TIE, D. magna, V. fischeri, seawater desalination facility

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924 Antibacterial Activities, Chemical Constitutes and Acute Toxicity of Peganum Harmala L. Essential Oil

Authors: Samy Selim

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Natural products are still major sources of innovative therapeutic agents for various conditions, including infectious diseases. Peganum harmala L. oil had wide range uses as traditional medicinal plants. The current study was designed to evaluate the antibacterial activity of P. harmala essential oil. The chemical constitutes and toxicity of these oils was also determined to obtain further information on the correlation between the chemical contents and antibacterial activity. The antibacterial effect of the essential oils of P. harmala oil was studied against some foodborne pathogenic bacteria species. The oil of plant was subjected to gas chromatography-mass spectrometry (GC/MS). The impact of oils administration on the change in rate of weight gain and complete blood picture in hamsters were investigated. P. harmala oil had strong antibacterial effect against bacterial species especially at minimum inhibitory concentration (MIC) less than 75.0 μg/ml. From the oil of P. harmala, forty one compounds were identified, and the major constituent was 1-hexyl-2-nitrocyclohexane (9.07%). Acute toxicity test was performed on hamsters and showed complete survival after 14 days, and there were no toxicity symptoms occurred. This study demonstrated that these essential oils seemed to be destitute of toxic effect which could compromise the medicinal use of these plants in folk medicine.

Keywords: analysis mass spectrometry, antibacterial activities, acute toxicity, chemical constitutes, gas chromatography, weight gain, Peganum harmala

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923 In vitro Evaluation of the Anti-Methanogenic Properties of Australian Native and Some Exotic Plants with a View of Their Potential Role in Management of Ruminant Livestock Emissions

Authors: Philip Vercoe, Ali Hardan

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Samples of 29 Australian wild natives and exotic plants were tested in vitro batch rumen culture system for their methanogenic characteristics and potential usage as feed or antimicrobial to enhance sustainable livestock ruminant production system. The plants were tested for their in vitro rumen fermentation end products properties which include: methane production, total gas pressure, concentrations of total volatile fatty acids, ammonia, and acetate to propionate ratio. All of the plants were produced less methane than the positive control (i.e., oaten chaff) in vitro. Nearly 50 % of plants inhibiting methane by over 50% in comparison to the control. Eremophila granitica had the strongest inhibitory effect about 92 % on methane production comparing with oaten chaff. The exotic weed Arctotheca calendula (Capeweed) had the highest concentration of volatile fatty acids production as well as the highest in total gas pressure among all plants and the control. Some of the acacia species have the lowest production of total gas pressure. The majority of the plants produced more ammonia than the oaten chaff control. The plant species that produced the most ammonia was Codonocarpus cotinifolius, producing over 3 times as much methane as oaten chaff control while the lowest was Eremophila galeata. There was strong positive correlation between methane production and total gas production as well as between total gas production and the concentration of VFA produced with R² = 0.74, R² = 0.84, respectively. While there was weak positive correlation between methane production and the acetate to propionate ratio as well as between the concentration of VFA produced and methane production with R² = 0.41, R² = 0.52, respectively.

Keywords: in vitro Rumen Fermentation, methane, wild Australian native plants, forages

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922 In silico and Toxicity Study of the Combination of Roselle (Hibiscus sabdariffa L.) and Garlic (Allium sativum L.) as Antihypertensive Herbs

Authors: Doni Dermawan

Abstract:

Hypertension is a disease with a high prevalence in Indonesia. The prevalence of hypertension in Indonesia is based on the Basic Health Research (Riskesdas) in 2013 which amounted to 25.8%. Medicinal plants have been widely used to treat hypertension including roselle (Hibiscus sabdariffa L.) and garlic (Allium sativum L.) by a mechanism as angiotensin converting enzyme (ACE) inhibitor. The purpose of this research is to analyze the in silico (molecular studies) of pharmacological effects and toxicity of roselle (Hibiscus sabdariffa L.) and garlic (Allium sativum L.) as well as a combination of both are used as antihypertensive herbs. The results of study showed that roselle (Hibiscus sabdariffa L.) and garlic (Allium sativum L.) have great potential as antihypertensive herbs based on the affinity and stability of active substances to specific receptor with a much better value than a of antihypertensive drugs (lisinopril). Toxicity values determined by the method of AST, ALT and ALP in which the three values obtained indicate the presence of acute toxic effects that need to be considered in determining the dose of the extract of roselle and garlic as antihypertensives.

Keywords: Allium sativum, antihypertensive, Hibiscus sabdariffa, in silico, toxicity

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921 Safety Risks of Gaseous Toxic Compounds Released from Li Batteries

Authors: Jan Karl, Ondrej Suchy, Eliska Fiserova, Milan Ruzicka

Abstract:

The evolving electromobility and all the electronics also bring an increase of danger with used Li-batteries. Li-batteries have been used in many industries, and currently many types of the batteries are available. Batteries have different compositions that affect their behavior. In the field of Li-battery safety, there are some areas of little discussion, such as extinguishing of fires caused by Li-batteries as well as toxicity of gaseous compounds released from Li batteries, transport or storage. Technical Institute of Fire Protection, which is a part of Fire Brigades of the Czech Republic, is dealing with the safety of Li batteries. That is the reason why we are dealing with toxicity of gaseous compounds released under conditions of fire, mechanical damage, overcharging and other emergencies that may occur. This is necessary for protection of intervening of fire brigade units, people in the vicinity and other envirnomental consequences. In this work, different types of batteries (Li-ion, Li-Po, LTO, LFP) with different kind of damage were tested, and the toxicity and total amount of released gases were studied. These values were evaluated according to their environmental hazard. FTIR spectroscopy was used for the evaluation of toxicity. We used a FTIR gas cell for continuous measurement. The total amount of released gases was determined by collecting the total gas phase through the absorbers and then determining the toxicants absorbed into the solutions. Based on the obtained results, it is possible to determine the protective equipment necessary for the event of an emergency with a Li-battery, to define the environmental load and the immediate danger in an emergency.

Keywords: Li-battery, toxicity, gaseous toxic compounds, FTIR spectroscopy

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920 Histopathological and Biochemical Evaluation of Hydroxyurea-Induced Hepato-Pulmonary Toxicity and Lymphoid Necrosis in Rats

Authors: Samah Oda, Asmaa Khafaga, Mohammed Hashim, Asmaa Khamis

Abstract:

Toxicity of hydroxyurea (HU), a treatment for certain tumors, polycythemia, and thrombocytosis, was evaluated in rats in one-month toxicity study. Sixty male albino rats were equally classified into four groups. Rats received daily oral gavage of HU in 0, 250, 500, and 750 mg/kg b.wt. Chemical and histopathological assessment of liver, lung, spleen, and bone marrow was performed at 10, 20, and 30 days of the experiment. No significant change was reported in alanine aminotransferase (ALT), aspartate aminotransferase (AST), globulin, and albumin/ globulin ratio during the experiment. Significant decreases in alkaline phosphatase (ALP) and total albumin were reported in rats received 500 and 750 mg/kg b.wt of HU. In addition, total cholesterol level increased significantly after 10 days; however, it significantly decreased after 20 and 30 days of the experiment. Moreover, hepatocytic vacuolation and necrosis with portal inflammatory infiltrates were reported along experimental periods. Pulmonary congestion, hemorrhage, interstitial mononuclear infiltration, peribronchitis, and bronchial epithelial necrosis were also reported. Severe lymphocytic necrosis in spleen and severe loss of hematopoietic cells and replacement with corresponding adipose tissue in bone marrow tissues was demonstrated. In conclusion, HU could be able to induce severe dose and time-dependent hepato-pulmonary toxicity and lymphoid depression in rats.

Keywords: hydroxyurea, hepato-pulmonary toxicity, lymphoid depression, histopathology

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919 Overview and Pathophysiology of Radiation-Induced Breast Changes as a Consequence of Radiotherapy Toxicity

Authors: Monika Rezacova

Abstract:

Radiation-induced breast changes are a consequence of radiotherapy toxicity over the breast tissues either related to targeted breast cancer treatment or other thoracic malignancies (eg. lung cancer). This study has created an overview of different changes and their pathophysiology. The main conditions included were skin thickening, interstitial oedema, fat necrosis, dystrophic calcifications, skin retractions, glandular atrophy, breast fibrosis and radiation induced breast cancer. This study has performed focused literature search through multiple databases including pubmed, medline and embase. The study has reviewed English as well as non English publications. As a result of the literature the study provides comprehensive overview of radiation-induced breast changes and their pathophysiology with small focus on new development and prevention.

Keywords: radiotherapy toxicity, breast tissue changes, breast cancer treatment, radiation-induced breast changes

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918 Adverse Effects of Natural Pesticides on Human and Animals: An Experimental Analysis

Authors: Abdel-Tawab H. Mossa

Abstract:

Synthetic pesticides are widely used in large-scale worldwide for control pests in agriculture and public health sectors in both developed and developing countries. Although the positive role of pesticides, they have many adverse toxic effects on humans, animals, and the ecosystem. Therefore, in the last few years, scientists have been searching for new active compounds from natural resources as an alternative to synthetic pesticides. Currently, many commercial natural pesticides are available commercially worldwide. These products are recommended for uses in organic farmers and considered as safe pesticides. This paper focuses on the adverse effects of natural pesticides on mammals. Available commercial pesticides in the market contain essential oils (e.g. pepper, cinnamon, and garlic), plant extracts, microorganism (e.g. bacteria, fungi or their toxin), mineral oils and some active compounds from natural recourses e.g. spinosad, neem, pyrethrum, rotenone, abamectin and other active compounds from essential oils (EOs). Some EOs components, e.g., thujone, pulegone, and thymol have high acute toxicity (LD50) is 87.5, 150 and 980 mg/kg. B.wt on mice, respectively. Natural pesticides such as spinosad, pyrethrum, neem, abamectin, and others have toxicological effects to mammals and ecosystem. These compounds were found to cause hematotoxicity, hepato-renal toxicity, biochemical alteration, reproductive toxicity, genotoxicity, and mutagenicity. It caused adverse effects on the ecosystem. Therefore, natural pesticides in general not safe and have high acute toxicity and can induce adverse effects at long-term exposure.

Keywords: natural pesticides, toxicity, safety, genotoxicity, ecosystem, biochemical

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917 Protective Effect of Aframomun chrysanthum Seed Aqueous Extract in Acetaminophen-Induced Liver Toxicity in Rats

Authors: N. Nwachoko, E. B. Essien, E. O. Ayalogu

Abstract:

Owing to the outbreak of different diseases and microbial resistance to some available drugs, proper identification, and evaluation of plants have been encouraged. There have been claims worldwide by the traditional system that some plants possessed medicinal properties. Plants and their components have been said to be source of large amount of drugs which comprise of distinct groups such as antispasmodics, anticancer and antimicrobials. Researchers have reported that chemicals in plants are responsible for the medicinal uses of plants. Thus this study evaluated the protective effect of Aframomun chrysanthum seed aqueous extract in acetaminophen-induced liver toxicity in rats. A suspension of 750 mg/kg acetaminophen was administered once every 72 hours to induce toxicity in the rats. Oral administration of 500, 1000 and 2000 mg/kg body weight of the extract and 100 mg/kg of silymarin (reference drug) were administered for 10 days. Biochemical analysis showed significant (p < 0.05) increase in the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT)and alkaline phosphatase (ALP)as well as the concentrations of albumin (ALB) and total bilirubin (T.B.) levels in rats administered with acetaminophen only. The levels of these parameters were significantly (p < 0.05) decreased in the groups pretreated with the extract.

Keywords: Aframomun chrysanthum, silymarin, hepatoprotective, toxicity

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916 Protective Effect of Celosia Argentea Leaf Extract on Cadmium Induced Toxicity and Oxidative Stress in Rats

Authors: Sulyman Abdulhakeem Olarewaju, S. O. Malomo, M. T. Yakubu, J. O. Akolade

Abstract:

The ameliorative effect of Celosia argentea var. cristata leaf extract against cadmium (Cd) induced oxidative stress and toxicity in selected tissues of rats was investigated. Toxicity coupled with oxidative stress was induced in rats by oral administration of Cd (8 mg/kg b. wt). Preliminary quantitative phytochemical and in vitro antioxidant analyses showed that the methanolic extract of C. argentea leaves was constituted by polyphenols (5.72%), saponins (3.20%), tannins (0.65%) and cadenolides (0.006%). IC50 of 9800, 7406, and 45.04 μg/ml were recorded for inhibition of linoleic acid oxidation, 2, 2-diphenyl-1-picrylhydrazyl and hydrogen peroxide radicals respectively. Simultaneous administration of C. argentea leaf extract with Cd significantly attenuated Cd-induced elevation of serum enzyme markers such as aspartate and alanine transaminase, alkaline and acid phosphatase as well as γ-glutaryltransferase in a dose-dependent fashion, while their reduced level in the liver were significantly increased. Higher levels of enzymatic antioxidants; superoxide dismutase and catalase activities were observed in the liver, brain, kidney and testes of the Cd-induced rats treated with C. argentea extract, while lipid peroxidation expressed in malondialdehyde concentrations were lower when compared to values in rats administered Cd only. Other Cd-induced toxicity and stress markers in the serum viz. reduced uric acid and albumin levels as well as elevated total and unconjugated bilirubin were attenuated by the extract and their values compared favorably with those animals co-administered cadmium with ascorbic acid. Data from the study showed that oral administration of extract from the leaf C. argentea may ameliorate Cd-induced oxidative stress and toxicity in rats.

Keywords: toxicity, cadmium, celosia, antioxidants, oxidative stress

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915 Analgesic, Toxicity and Anti-Pyretic Activities of Methanolic Extract from Hyoscyamus albus Leaves in Albinos Rats

Authors: Yahia Massinissa, Henhouda Affaf, Yahia Mouloud

Abstract:

The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by brewer’s yeast induced fever in rats. For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of Hyoscyamus albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of Hyoscyamus albus was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, methanolic extract, toxicity, analgesic activity, antipyretic activity, formalin test

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914 Studies on Toxicity and Mechanical Properties of Nonmetallic Printed Circuit Boards Waste in Recycled HDPE Composites

Authors: Shantha Kumari Muniyandi, Johan Sohaili, Siti Suhaila Mohamad

Abstract:

The aim of this study was to investigate the suitability of reusing nonmetallic printed circuit boards (PCBs) waste in recycled HDPE (rHDPE) in terms of toxicity and mechanical properties. A series of X-ray Fluorescence Spectrometry (XRF) analysis tests have been conducted on raw nonmetallic PCBs waste to determine the chemical compositions. It can be seen that the nonmetallic PCBs approximately 72% of glass fiber reinforced epoxy resin materials such as SiO2, Al2O3, CaO, MgO, BaO, Na2O, and SrO, 9.4% of metallic materials such as CuO, SnO2, and Fe2O3, and 6.53% of Br. Total Threshold Limit Concentration (TTLC) and Toxicity Characteristic Leaching Procedure (TCLP) tests also have been done to study the toxicity characteristics of raw nonmetallic PCB powders, rHDPE/PCB and virgin HDPE for comparison purposes. For both of the testing, Cu was identified as the highest metal element contained in raw PCBs with the concentration of 905 mg/kg and 59.09 mg/L for TTLC and TCLP, respectively. However, once the nonmetallic PCB was filled in rHDPE composites, the concentrations of Cu were reduced to 134 mg/kg for TTLC and to 3 mg/L for TCLP testing. For mechanical properties testing, incorporation of 40 wt% nonmetallic PCB into rHDPE has increased the flexural modulus and flexural strength by 140% and 36%, respectively. While, Izod Impact strength decreased steadily with incorporation of 10 – 40 wt% nonmetallic PCBs.

Keywords: nonmetallic printed circuit board, recycled HDPE, composites, mechanical properties, total threshold limit concentration, toxicity characteristic leaching procedure

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913 Comparative and Combined Toxicity of NiO and Mn₃O₄ Nanoparticles as Assessed in vitro and in vivo

Authors: Ilzira A. Minigalieva, Tatiana V. Bushueva, Eleonore Frohlich, Vladimir Panov, Ekaterina Shishkina, Boris A. Katsnelson

Abstract:

Background: The overwhelming majority of the experimental studies in the field of metal nanotoxicology have been performed on cultures of established cell lines, with very few researchers focusing on animal experiments, while a juxtaposition of conclusions inferred from these two types of research is blatantly lacking. The least studied aspect of this problem relates to characterizing and predicting the combined toxicity of metallic nanoparticles. Methods: Comparative and combined toxic effects of purposefully prepared spherical NiO and Mn₃O₄ nanoparticles (mean diameters 16.7 ± 8.2 nm and 18.4 ± 5.4 nm respectively) were estimated on cultures of human cell lines: MRC-5 fibroblasts, THP-1 monocytes, SY-SY5Y neuroblastoma cells, as well as on the latter two lines differentiated to macrophages and neurons, respectively. The combined cytotoxicity was mathematically modeled using the response surface methodology. Results: The comparative assessment of the studied NPs unspecific toxicity previously obtained in vivo was satisfactorily reproduced by the present in vitro tests. However, with respect to manganese-specific brain damage which had been demonstrated by us in animal experiment with the same NPs, the testing on neuronall cell culture showed only a certain enhancing effect of Mn₃O₄-NPs on the toxic action of NiO-NPs, while the role of the latter prevailed. Conclusion: From the point of view of the preventive toxicology, the experimental modeling of metallic NPs combined toxicity on cell cultures can give non-reliable predictions of the in vivo action’s effects.

Keywords: manganese oxide, nickel oxide, nanoparticles, in vitro toxicity

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912 Toxicological Analysis of Some Plant Combinations Used for the Treatment of Hypertension by Lay People in Northern Kwazulu-Natal, South Africa

Authors: Mmbulaheni Ramulondi, Sandy Van Vuuren, Helene De Wet

Abstract:

The use of plant combinations to treat various medical conditions is not a new concept, and it is known that traditional people do not only rely on a single plant extract for efficacy but often combine various plant species for treatment. The knowledge of plant combinations is transferred from one generation to the other in the belief that combination therapy may enhance efficacy, reduce toxicity, decreases adverse effects, increase bioavailability and result in lower dosages. However, combination therapy may also be harmful when the interaction is antagonistic, since it may result in increasing toxicity. Although a fair amount of research has been done on the toxicity of medicinal plants, there is very little done on the toxicity of medicinal plants in combination. The aim of the study was to assess the toxicity potential of 19 plant combinations which have been documented as treatments of hypertension in northern KwaZulu-Natal by lay people. The aqueous extracts were assessed using two assays; the Brine shrimp assay (Artemia franciscana) and the Ames test (Mutagenicity). Only one plant combination (Aloe marlothii with Hypoxis hemerocallidea) in the current study has been previously assessed for toxicity. With the Brine shrimp assay, the plant combinations were tested in two concentrations (2 and 4 mg/ml), while for mutagenicity tests, they were tested at 5 mg/ml. The results showed that in the Brine shrimp assay, six combinations were toxic at 4 mg/ml. The combinations were Albertisia delagoensis with Senecio serratuloides (57%), Aloe marlothii with Catharanthus roseus (98%), Catharanthus roseus with Hypoxis hemerocallidea (66%), Catharanthus roseus with Musa acuminata (89%), Catharanthus roseus with Momordica balsamina (99%) and Aloe marlothii with Trichilia emetica and Hyphaene coriacea (50%). However when the concentration was reduced to 2 mg/ml, only three combinations were toxic which were Aloe marlothii with Catharanthus roseus (76%), Catharanthus roseus with Musa acuminata (66%) and Catharanthus roseus with Momordica balsamina (73%). For the mutagenicity assay, only the combinations between Catharanthus roseus with Hypoxis hemerocallidea and Catharanthus roseus with Momordica balsamina were mutagenic towards the Salmonella typhimurium strains TA98 and TA100. Most of the combinations which were toxic involve C. roseus which was also toxic when tested singularly. It is worth noting that C. roseus was one of the most frequently used plant species both to treat hypertension singularly and in combination and some of the individuals have been using this for the last 20 years. The mortality percentage of the Brine shrimp showed a significant correlation between dosage and toxicity thus toxicity was dosage dependant. A combination which is worth noting is the combination between A. delagoensis and S. serratuloides. Singularly these plants were non-toxic towards Brine shrimp, however their combination resulted in antagonism with the mortality rate of 57% at the total concentration of 4 mg/ml. Low toxicity was mostly observed, giving some validity to combined use, however the few combinations showing increased toxicity demonstrate the importance of analysing plant combinations.

Keywords: dosage, hypertension, plant combinations, toxicity

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