Short Communications 2018-03-28T11:49:19+00:00

12th SCT

April 18-21, 2018. Belgrade, Serbia

Short Communications

The Use of Selected Marine Organisms in Nanoecotoxicology: Experiments on Mussels, Sea Urchins and Crabs


Petra Burić1, Maja Levak1, Lorena Perić2, Ines Kovačić3, Dijana Pavičić-Hamer1, Daniel Mark Lyons1

1 Laboratory for Marine Nanotechnology and Biotechnology, Center for Marine Research,     Ruđer Bošković Institute, Rovinj, Croatia, 2 Laboratory for Aquaculture and Pathology of Aquatic Organisms, Division for Marine and   Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia, 3Juraj Dobrila University of Pula, Pula, Croatia


With the growing use of nanoparticles in a broad range of industrial and consumer applications there is an increasing likelihood that such nanoparticles will enter the aquatic environment and be transported through freshwater systems, eventually reaching marine waters. Due to silver’s known antimicrobial properties and widespread use of silver nanoparticles (AgNP) in consumer products, the latter’s environmental fate and impact is therefore of particular concern. In this context, we have investigated the effect of low, environmentally-relevant, concentrations (from 1 up to 1000 µg L-1) and different sizes of AgNP (10, 20, 40, 60 and 100 nm) in selected marine invertebrates: sea urchins embryos (Arbacia lixula and Paracentrotus lividus), mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii). The results have showed that 50-1000 µg L-1 AgNP have a negative effect on the sea urchin embryonal development, fertilisation success and have cytogeneotoxic potential toward sea urchin embryos. This findings were dependent on the investigated sea urchin species and the size of nanoparticles used. Mussels were sensitive to 50 µg L-1 AgNP (60 nm) showing a stress response at the entire organism level, destabilisation of hemocites lysosomal membranes, elevation of enzymatic glutathione S-transferase activity and metallothionein content in mussel tissue. However, no effect was observed in the activity of acetylcholinesterase after AgNP treatment. AgNP concentrations from 100-1000 µg L-1 (60 nm) caused mortality in crabs, while up to 500 µg L-1 caused increased total hemocyte numbers. Thus, the selected marine organisms have shown to be useful models for investigating the toxicity of nanoparticles.


Keywords: silver nanoparticle, embryo, lysosomal membrane stability, glutathione S-transferase, metallothionein



The Reference Value for Biomonitoring in Chemicals Risk Area in Thailand


Nalinee  Sripaung, Ph.D.

Bureau of Occupational and Environmental Diseases, DDC, Ministry of Public Health, Thailand



The workers’ biological standard value (WBSV) is generally used to assess chemicals health risks in community. It may cause the deviate biomonitoring. Therefore, this study was aimed to set the pilot reference value for exposed people to chemicals. The study was divided into two phases. Phase I was proceeded in one industrial zone in Thailand during the year 2012 – 2014 to collect blood and urine samples of 402 working – age people in community accompanied with in-depth interview. The 4 heavy metals (arsenic, cadmium, lead, mercury) in blood and urine were analyzed by AAS – analysis and the metabolites of 4 VOCs (benzene, toluene, styrene, xylenes) in urine were analyzed by HPLC – analysis. The volunteers’ 8 average chemicals concentration (VACC) were calculated. Phase II was proceeded during the year 2015 – 2016 to compare risk group identification between usage of VACC and usage of WBCV. The results were presented in percentage. The results showed approximate 90 percent of VACC were lower than WBSV. The exceptional result was volunteers’ average urinary arsenic concentration. It was clearly higher than WBSV. The comparative results showed the adjusted amount of risk people by VACC was higher than the adjusted amount risk people by WBSV. This study indicated that general people’s average chemicals concentration should be used as the reference value for biomonitoring and active health surveillance. Besides, daily intake of chemical mixtures in food chain should be concerned.

Keywords:  reference value, biomonitoring, chemicals health risks, exposed people, active health surveillance



Evaluation of Manganese, Copper, Zinc and Selenium Levels in Patients with Primary Epithelial Ovarian Cancer


Aydan Caglayan1, Doruk Cevdi Katlan2, Zafer Selçuk Tuncer3, Kunter Yüce3

1Hacettepe University, Faculty of Pharmacy, Department of Toxicology, Ankara, Turkey

2Suleymaniye Research and Education Hospital, Obstetrics and Gynecology, İstanbul, Turkey

3Hacettepe University, Faculty of Medicine, Department of Obstetrics and Gynecology, Ankara , Turkey

Correspondence: or

Ovarian cancer is the fifth most frequent cause of cancer deaths in women in European Union countries whereas seventh among all cancers and second among gynecological cancers in Turkey. Epithelial ovarian tumors (EOC) comprises 90% of malignant, 60% of all ovarian tumors, and 75% of EOC are characterized by serous histologic subtype. EOC has been associated with oxidative stress due to epithelial inflammation which makes ovaries more vulnerable to the deleterious effects of reactive oxygen species (ROS). However, antioxidant enzymes (AOEs) [manganese-superoxide dismutase (Mn-SOD), copper,zinc-superoxide dismutase (Cu,Zn-SOD), catalase (CAT) and glutathione peroxidase (GPx1)] protect cells against the biological damage of ROS and support cancer prevention by maintaining normal cell cycle progression, inducing apoptosis, and inhibiting proliferation/ tumor invasion/ angiogenesis/ inflammation.

In the present study, we aimed to measure the trace elements [manganese (Mn), copper (Cu), zinc (Zn) and selenium (Se)] which are structurally/functionally associated with the AOEs by inductively coupled plasma/mass-spectrometry (ICP/MS) in blood samples of patients with EOC (M, n= 30) and compare the data with healthy subjects (C, n=40). Serous EOC (M1, n=18) data were also evaluated according to the tumor grading [well or moderately well differentiated (G1-2) vs. poorly differentiated or undifferentiated (G3)] and disease staging [stage I-II (SI-II) vs. stage III (SIII)]. We obtained; i) The Mn and Se levels of M were significantly lower (∼1.4 and ~1.2 times) than C, ii) only Mn levels were changed [(G3(Mn) < G1-2(Mn), 32%] in M1, iii) significant correlations were observed between [Cu and Zn levels (r=0.701, p=0.036) in G1-2 and (r=0.686, p=0.041) in G3; Cu and Se levels (r=0.960, p=0.000) in G3; Mn levels and Mn-SOD expression (r= 0.551, p=0.006) in M, (r=0.857, p=0.007) in G1-2 and (r= 0.690, p=0.056) in G3; Se levels and erythrocyte GPx1 activity (r=0.660, p=0.053) in G1-2, Se levels and erythrocyte Cu,Zn-SOD activity (r=0.693, p=0.038) in G3]. The study revealed that the trace elements particularly Mn and Se may be of value in the EOC and although Mn level is important in terms of discriminating the tumor grades, mutual correlations between Cu-Zn and Cu-Se levels are remarkable in all grades. Morever, Se is the major important trace element correlates significantly with the eythrocyte AOEs.

Keywords: epithelial ovarian cancer, manganese, copper, zinc, selenium


Acute Poisoning in Children in Azerbaijan


Ismayil Afandiyev

Azerbaijan Medical University, Azerbaijan, Baku


Acute poisoning in children is one of the leading causes of morbidity and mortality in many countries. Data on the epidemiology of pediatric poisoning in Azerbaijan is very scarce. In this observation study, all children poisonings over eight years period (1st January 2009 to 31st December 2016) admitted to Poison Center (PC) in Baku, Azerbaijan were reviewed using electronic and paper medical records. There were a total 2949 pediatric admissions to PC (17.9% of all poisoning cases).  Among them only 6.2% were intentional poisonings. One thousand six hundred fifty-two patients (56.2%) were male. Pharmaceuticals (n=1276, 43.3%), organic solvents/oil products (n=562, 19.1%), and corrosive substances (n=380, 12.9%) were the most common agents responsible for poisoning in children. Thirty fatal cases reported during the study period. Mortality in children (1.02%) was significantly lower (p<0.05) than mortality in adult group (2.56%). Adoption of educational prevention program for parents as well as enforcement child-resistant packaging and reducing toxicity of household chemicals can decrease poisoning cases among children.

Key words: children poisoning, epidemiology, Azerbaijan



Toxic Effects of Bisphenol Analogues: Are Alternatives Safe?


Sheikh Raisuddin1 Shikha Sharma1, and Jasim Khan 1

Department of Medical Elementology & Toxicology, Jamia Hamdard (Hamdard University), New Delhi 110062, India

E-mail address:

There is enough literature on bisphenol A (BPA), as it is one of the most studied bisphenol analogues and its production is bulk. Due to sustained media coverage, there is public awareness about the hazardous nature of BPA. This has led to search for alternatives (substitutes) to BPA. Over a dozen bisphenol analogues have been in market. Safety data on these substitutes are limited. However, these analogues appear to be active and available literature indicates their propensity to bind with molecular targets. We conducted a comprehensive in silico study on the binding efficiency of selected bisphenol analogues with their likely target peroxisome proliferator-activated receptors (PPAR). We selected mouse PPARα. Upon interaction with PPARα BPA and its halogenated analogues showed binding energy scores (Kcal/Mol) in the order: tetrabromobisphenol A (-8.75) ˃bisphenol Z (-8.09) ˃ BPC = tetrachlorobisphenol A (-7.77) ˃ bisphenol E (-7.24) ˃ bisphenol B (-7.15) ˃ bisphenol F (7.02) ˃ bisphenol S (6.02) ˃ BPA (5.60) ˃ bisphenol AF (-5.39). Lowest binding energy score of positive control GW 7647 was -7.89. Certain BPA analogues showed higher binding efficiency with PPARα than BPA. In silico results suggest that some of the halogenated analogues of BPA have strong binding affinity with PPARα. Taking bisphenol S (BPS) in vivo study in mice showed hepatotoxic effects of BPS which included lipid peroxidation in a dose-dependent manner along with mild histopathological changes. These results reveal that a proper risk assessment profiling is desirable to ensure safety aspects of BPA alternatives.

Keywords: endocrine disrupting chemicals, nuclear receptors, binding, bisphenol A, bisphenol analogues.



Endocrine Disrupting Activity in Sewage Sludge: Screening Method and Cost-effective Strategy for Detoxification


Dânia E. C. Mazzeo 1,2, Andrea Misovic 2,  Maria A. Marin-Morales 3, Mary Rosa R. Marchi 1, Jörg Oehlmann 2

1 Department of Analytical Chemistry, Institute of Chemistry, UNESP — Univ Estadual Paulista, Araraquara, SP, Brazil, 2 Department Aquatic Ecotoxicology, Goethe University Frankfurt am Main, Germany, 3 Department of  Biology, Institute of Bioscience, UNESP — Univ Estadual Paulista , Rio Claro, SP, Brazil


Sewage sludge (SS) presents a high agronomic potential due to the presence of organic matter, macro and micronutrients, encouraging its recycling in agriculture as a soil conditioner. However, the presence of toxic substances can preclude this practice. Among these substances, the endocrine disrupting chemicals (EDCs) require special attention since they can potentially interfere with the hormonal system of exposed organisms, resulting in metabolic and reproductive disorders, even when present in low concentrations. To enable a safe disposal of this waste in agriculture, this study verified the estrogenic, androgenic and retinoic-like activities of anaerobic SS samples by means of yeast-based reporter-gene assays. Additionally, aiming to eliminate the EDCs from SS, a detoxification process combining microorganisms and biostimulating agents (soil, sugarcane bagasse, and coffee grounds) was performed for periods of 2, 4 and 6 months. A fractionation procedure of samples, dividing the target sample extract into several fractions according to their polarity, was conducted in order to decrease the matrix complexity. Before the detoxification process, the studied SS induced significant agonistic activity at the human estrogen receptor α (hERα), androgen receptor (hAR) as well as the retinoic acid receptor (RARα) but not at the retinoid X receptor (RXRα), in, at least, one of the tested fractions. However, after the SS detoxification, no significant activities were observed, showing that the technology applied here was efficient to eliminate receptor-mediated toxicity. Moreover, the recombinant yeast assay and the fractionation procedure proved to be valuable methods to easily detect EDCs in SS.

Financial support: FAPESP-BEPE Process 2017/10198-3


Keywords: endocrine disruptors, receptor-mediated toxicity, in vitro bioassay, chemical fractionation, biostimulation



Incidence of Enrofloxacin its Primary Metabolite and Chlortetracycline Residues in Eggs and Broiler Meat from Tamilnadu, India

Ghadevaru Sarathchandra

Pharmacovigilance Laboratory for Animal Feed and Food Safety, Centre For Animal Health Studies, TANUVAS



The administration of fluoroquinolones and Tetracyclines to food animals without an adequate withdrawal time may lead to violative concentrations of residues in foods destined for human consumption. To ensure control over the presence of antibiotic residues in food stuffs of animal origin, European Union and Japan have set maximum residue limits for antibiotic residues in edible animal tissues. Since 2005 the U.S. Food and Drug Administration has banned the use of enrofloxacin in poultry/food animals. In view of this, the prevalence of enrofloxacin and its primary metabolite ciprofloxacin residues in broiler meat and organ samples of field origin was explored in the present study. A total of 180 numbers of broiler chicken edible tissue samples (liver, kidney, breast muscle, thigh muscle and skin each 36 numbers) were randomly purchased from various retail outlets in Chennai, India. Sampling was carried out as per the guidelines prescribed by Codex Alimentarius). Liver, kidney, muscle , skin and egg samples were extracted and subjected to enrofloxacin and ciprofloxacin quantification by validated High Performance Thin Layer Chromatography (HPTLC) – Fluorescent Densitometry.

Liver (20%), kidney (15%) and skin (10.%) samples had enrofloxacin residues higher than the MRLs and found to be violating the regulations of Japan. Indeed, all muscle and egg samples were found to be safe as per EU and Japan MRLs This study therefore stresses the need for adhering the withdrawal period as prescribed by various regulatory bodies (FDA, EU and Japan) and the need for stringent regulation for the use of antimicrobial drugs in the poultry industry as well as the inspection of chicken for antimicrobial residues prior to marketing in India.

Keywords: veterinary drug residue, HPTLC, food safety

Antioxidant Potentials of Cedrelopsis grevei Leave Extract against Chlorpyrifos Induced Oxidative Stress in Kidney of Male Rats


Tarek Heikal

Narcotics, Ergogenics and Poisons Dept, Medical Research Division, National research Centre (NRC), Cairo, Egypt


In the present study, the protective effect of Cedrelopsis grevei (C. grevei) leave extracts against renal oxidative damage and nephrotoxicity induced by chlorpyrifos (CPF) in male rats was undertaken. Four groups containing six rats each were selected. Group I served as control. Groups II rats were received 300 mg/kg extracts by oral gavages. Groups III rats were given a single daily oral doses of CPF (13.5 mg kg-1 kg-1, 1/10 LD50, in corn oil) for 28 consecutive days. Groups V rats were simultaneously given the same doses of extract and CPF as in groups II and III, respectively. In CPF-treated rats, significant reduction in body weight and elevation in kidney weight were observed compared with control. Also, significant perturbations of renal function as evidenced via increase in blood urea nitrogen (BUN) and serum creatinine level and oxidative damage as evidenced via augmentation in kidney lipid peroxidation (LPO) as well as depletion in kidney antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) in CPF-treated rats in comparing with control. Histopathological analysis of the kidney revealed that supplementation with C. grevei resulted in nil to mild vacuolization, swelling and degeneration in the endothelium of glomerular tuft and the epithelium of lining tubules. In conclusion, the use of C. grevei extract appeared to be beneficial to rats, to a great extent by attenuating and restoring the damage sustained by insecticide exposure.

Keywords: antioxidants, chlorpyrifos, kidney, oxidative stress, renal toxicity



Toxicological Assessment of Aqueous Leaf Extract of Bryophyllum pinnatum on Body and Organ Weights, and Haematologic Parameters in Rodents


Omoniyi Yemitan 1, Akinsuyi Akinsegun 2, Sunday Olayemi 34

1 Department of Pharmacology, Therapeutics & Toxicology, Lagos State University College of Medicine, Ikeja, Lagos, Nigeria, 2,3 Department of Pharmacology, Therapeutics & Toxicology, College of Medicine of the University of Lagos, Idi-Araba, Lagos, Nigeria.


The acute and subchronic toxicities of the aqueous leaf extract of Bryophyllum pinnatum (ALBP) was studied on the body and vital organs and haematologic parameters in rodents, with a view to predicting its safety in humans.

To five groups of albino mice (n = 5), acute oral or i.p. 24h -14 days toxicity was tested for ALBP, up to 5000 mg/kg, then observed for morphologic changes and mortalities. For the 90-day subchronic study, male rats (16/group) received daily 10, 100, 1000 mg/kg extract doses or distilled water (control); body weights were measured weekly; after 90 days, some rats were sacrificed (10/group) for vital organs weights and haematology. Remaining rats (6/group) were retained, untreated, for reversibility study.

Oral ALBP caused sedation, tachycardia and hypercapnia; intraperitoneal doses produced LD50 of 1650 mg/kg. Subchronic test caused significant (P<0.05) weight reduction. At 1000 mg/kg, ALBP significantly caused increased weights of kidneys, liver and spleen, but reduction in weights of lungs and testes. Reversal of weights of the liver and spleen, but not the lungs and testes, was recorded. At 100 mg/kg, increases in PCV % & WBC, but at 1000 mg/kg, significant reduction of RBC, Hb concentration, PCV % and platelets were recorded. For WBC, significant and irreversible elevation was recorded at 100 & 1000 mg/kg of ALBP.

At high doses, ALBP has potential to cause toxic effects on lungs and testes, as well as anaemia, thereby monitoring of during long-term use is required.

Keywords: Bryophyllum pinnatum, toxicity, body weight, testicular toxicity, anaemia



Occupational Exposure Modeling of Pesticides:  State of the Art and Challenges for their Implementation in Developing Countries


Wells Utembe1, Mary Gulumian1,2

1 Toxicology Department, National Institute for Occupational Health, South Africa, 2 School of Pathology, University of the Witwatersrand, South Africa



Occupational exposure to pesticides is determined by factors including weather, level of protection, nature and duration of task, state of formulation, application equipment, type of crop and behaviour of the pesticide handler.  Occupational exposure modelling is therefore a very complex and arduous task that is often achieved through the use of empirical models (derived from regional or local experimental data). Various occupational exposure modelling tools were therefore reviewed to assess if they can be utilized in the risk assessment of pesticides in developing countries and also to evaluate the challenges that are encountered when utilizing these tools.

Examples of exposure modelling tools reviewed include the Pesticide Handlers Exposure Database (PHED) (North America), the German Biologische Bundesanstalt für Land- und Forstwirtschaft (BBA), United Kingdom Predictive Operator Exposure Model (UK-POEM), the European Predictive Operator Exposure Model (EUROPOEM), and the Dutch model all of which were developed in industrialised countries.

A review of these various occupational exposure modelling tools to assess if they can also be utilized in developing countries have revealed that each one of these models requires a number input parameters that may not be readily available in developing countries. Most importantly, as empirical models, existing pesticide occupational exposure models are only applicable in domains similar to those for which they were developed. It is therefore proposed that their applicability in some countries cannot be assumed, and therefore recommended that their validation and recalibration may be required.

Keywords:  occupational exposure, modelling, risk assessment

Immunomodulatory Properties of Zinc Oxide


Pasqualla Fagundes dos Santos 1, 2, Kathrin Becker 1, Harald Schennach 2, Dietmar Fuchs 1, Johanna M Gostner 3

1Division of Biological Chemistry, Medical University of Innsbruck, Austria, 2Central Institute of Blood Transfusion and Immunology, University Hospital, Innsbruck, Austria, 3Division of Medical Biochemistry, Medical University of Innsbruck, Austria


Zinc oxide (ZnO) nanomaterials are used not only in technical and chemical industry, but also pharmaceutical, food and cosmetic applications are frequent, due to the relatively low toxicity and biodegradability. While the use of ZnO and nanomaterials is considered not to pose a risk of adverse effects after dermal exposure, there are concerns regarding ZnO containing aerosols.

Human peripheral blood mononuclear cells (PBMC) have been applied successfully to investigate immunomodulatory effects of compounds and materials. For such studies, the interferon-gamma-induced metabolic pathways of tryptophan breakdown to kynurenine via indoleamine 2,3-dioxygenase (IDO-1), and neopterin formation via GTP-cyclohydrolase can be used as readout.

ZnO bulk material and nanoparticles were analysed in this in vitro setting in a concentration range from 2.3 to 37.5 µg/ml. Nanoparticles and bulk materials showed different toxicity, the nanoparticles reducing cell viability in a concentration-dependent manner up to 70% with the highest concentration, at which the first effects of the bulk material became obvious. The kynurenine to tryptophan ratio, a measure of IDO-1 activity, decreased with all materials dose-dependently in mitogen-stimulated cells. However, nanomaterial treatment of unstimulated cells increased IDO-1 activity, while this effect was less prominent with the bulk treatment. Neopterin concentrations showed the same trend.

Data indicate that within a certain concentration range, ZnO nanoparticles may have some activating effect on unstimulated PBMC, while in stimulated cells and with higher concentrations immunosuppressive effects prevail. A closer elucidation of these effects is clearly warranted.

Keywords: ZnO nanoparticles, in vitro setting, immunosuppressive effects

Combined Effects of BEA and ENB on Jurkat T-cells at the Transcriptomic Level

Lara Manyes , Laura Escrivá , María José Ruiz , Manuel Alonso

Laboratory of Food Chemistry and Toxicology, Faculty of Pharmacy, University of Valencia, Spain.


Beauvericin (BEA) and enniatins (ENs) are mycotoxins produced by various Fusarium species. BEA produces cytotoxicity, intracellular reactive oxygen species and lipid peroxidation and EN B is an ionophore antibiotic. These mycotoxins are normally concomitant in cereal-based products. Even if both able to reach bloodstream, luckily they show poor bioavailability. The study of transcriptional changes provoked by the individual treatment of BEA on Jurkat T-cells (1,5-3-5 µM during 24, 48 and72 h), showed a large number of differentially expressed genes mainly related to respiratory chain, apoptosis, and caspase cascade activation. When these cells were exposed to EN B at the same time and concentration, pathways affected were electron transport chain and oxidative phosphorylation. The aim of this work is to study the transcriptional changes in Jurkat T-cells after exposure to the combination of BEA and EN B. Firstly, a bioinformatics approach was conducted. The comparison between the differentially expressed genes (DEGs) from individual mycotoxin exposure (BEA= 5719 and EN B=5750) show coincidences in 2693 DEGs. Moreover, from 96 genes related to the respiratory chain, 32 are DEGs in both individual exposures. Other identical DEGs found were related to caspase activation, apoptosis and programmed cell death. Interestingly, only 14 DEGs were found for both treatments and for all conditions belonging 11 of them to mitochondrial genome. Secondly, RT-PCR confirmation was carried out. In conclusion, the results point to mitochondrial toxicity of BEA and EN B through electron transport chain inhibition and oxidative phosphorylation disruption.


Keywords: mitochondria, mycotoxin, oxidative phosphorylation, respiratory chain, PCR



Evaluation of Nephrotoxic Effects of Zearalenone in Human Kidney Cells


Ecem Fatma Karaman1, Ileyna Arıman1, Sibel Ozden1

1 Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Istanbul University, 34116-Beyazit, Istanbul, Turkey.


Zearalenone (ZEA), produced by various Fusarium species, is a non-steroidal estrogenic mycotoxin. It contaminates cereals such as corn, wheat, oat and soybean resulting in human and animal consumption. Therefore, it has serious health hazards to humans and animals. ZEA causes severe reproductive toxicity in humans on account of its xenoestrogenic activity. However, toxicity mechanism of ZEA has not been elucidated yet. We investigated dose-dependent effects of ZEA (0, 1, 10 and 50 µM for 24 h) in generation of reactive oxygen species (ROS), cell cycle regulation, global DNA methylation and gene regulation related with oxidative damage in human kidney (HK-2) cells. IC50 value of ZEA was determined as 151.27 µM and 60.5 µM in HK2 cells for 24 h by MTT and NRU tests, respectively. ZEA increased ROS levels in dose-dependent manner. BrdU test showed that 50 µM of ZEA treatment altered  cell proliferation significantly. ZEA slightly increased levels global DNA methlyation and gene expression levels of related DNMT1 and MGMT enzymes. In addition, gene expression levels of IL6, IL8, TNFα, MAPK8, NF-κB1, HO1, α-GST, HSP70, Nrf2, L-FABP genes were investigated using real time PCR in response to ZEA. We observed dramatically increase on gene expression levels of α-GST and HSP70 confiming that protein expression levels of HSP70 also increased by western blot analysis. We suggested that HSP70 may be a key molecular biomarker in the nephrotoxicity of ZEA. Our study might provide a new perspective in toxicity mechanism of ZEA in kidney cells.

Keywords: zearalenone, DNA methylation, odidative damage, nephrotoxicity, HK-2 cells


Levels of Heavy Metals and Ochratoxin A in Medicinal Plants Commercialized in Turkey


Hakan Ozden1, Sibel Ozden2

1Division of Botany, Department of Biology, Faculty of Science, Istanbul University, 34134, Suleymaniye, Istanbul, Turkey, 2Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Istanbul University, 34116 Beyazit, Istanbul, Turkey.


The aim of this study as to determine the levels of lead, cadmium and ochratoxin A (OTA) in frequently used medicinal plants. Totally twenty-one samples of linden, chamomile and sage were obtained during the spring and summer period of the year 2016 from local markets and traditional bazaars in Istanbul, Turkey. Microwave-assisted digestion was applied for the preparation of the samples and inductively coupled plasma technique with optical emission spectrometry (ICP-OES) was used for the determination of lead and cadmium. Determination of OTA was carried out using high performance liquid chromatography with fluorescence detector (HPLC-FLD) after immunoaffinity column clean-up. OTA was detected in only one chamomile sample with a low concentration level of 0.034 µg/kg. According to the results of ICP-OES analysis, lead in the concentration range of 4.125-6.487 mg/kg, 3.123-5.769 mg/kg and 3.229-5.985 mg/kg and cadmium in the concentration range of 0.324-0.524 mg/kg, 0.365-0.51 mg/kg and 0.321-0.474 mg/kg was found in linden, chamomile and sage teas, respectively. We indicated that levels of Pb and OTA were found below the maximum permissible level whereas high levels of Cd were observed in medicinal plants which may not pose health risk for the consumers according to the exposure assessment. However, it is suggested that other mycotoxins and heavy metal content should be carefully considered in medicinal plants.


Keywords: lead, cadmium, ochratoxin A, linden, chamomile, sage



Manganese-induced Brain Mitochondrial Dysfunction is Associated with Impaired Motor Functions in Rats: Protection with Nutrient Metal Mixture Supplementation


1Chand Basha Davuljigari, 2Katari Sudheer, 2Umamaheswari Amineni, 1Sreenivasulu Reddy Motireddy, 1Rajarami Reddy Gottipolu.

1Department of Zoology, Sri Venkateswara University, Tirupati, India-517 502. 2Bioinformatics Centre, Department of Bioinformatics, Sri Venkateswara Institute of Medical Science University, Tirupati, India-517 507.


Exposure to excessive levels of manganese (Mn) results in a movement disorder which resembles Parkinson’s disease. The pathogenic mechanisms underlying its action are not elucidated. To determine the role of mitochondrial energy metabolism and oxidative stress in Mn-induced neurobehavioral dysfunctions, male pups were lactationally exposed to Mn (6 mg/Kg body weight) through intraperitoneal injection for a period of two weeks (5 days/week) from PND 15 to PND28. To study the protective effect of nutrient metal mixture, pups were administered calcium (Ca), iron (Fe) and zinc (Zn) in combination as 0.02% by a single gavage together with Mn injection. The results showed mitochondrial succinate dehydrogenase (SDH), lactate dehydrogenase (LDH), and isocitrate dehydrogenase (ICDH) activities decreased in cortex and cerebellum at PND 28, PND 60 and 3 months age group rats following exposure to Mn. Most notably, Mn exposure decreased the activities of thyoredoxin reductase (TrxR), aconitase (Acon), superoxide dismutase (SOD), and catalase (CAT) while the MDA levels increased in the cortex, and cerebellum of selected age groups of rats. In silico findings revealed that aconitate hydratase in complex with modified control cluster (S4F3Mn) influences the Acon activity in the presence of the substrate. Mn-exposed rats exhibited deficits in total locomotor activity and grip strength in rats. However, supplementation of the nutrient metal mixture containing Ca, Fe and Zn reversed the effects of Mn on energy metabolism, oxidative damage of mitochondria and motor behaviour of the rats. In conclusion, our findings demonstrate that exposure to Mn during the development of brain greatly increased the mitochondrial dysfunction, subsequently, associated with motor coordination deficits in rats. Furthermore our results suggest that application of nutrient metal mixture may potentially be beneficial in treating Mn- neurotoxicity.

Supported by Science and Engineering Research Board (DST, Govt. of India), File No. YSS/2015/000289.

Keywords: oxidative damage, in silico studies, aconitase, manganese, motor functions



OECD Country’s Research Productivity on Nanotoxicity Research: a Bibliometric Analysis


Mahmoud Abudayyak

Department of Pharmaceutical Toxicology, Faculty of Pharmacy, karadeniz technical University, Trabzon, Turkey


The superior properties of nanomaterials have dramatically risen the usage of these materials in various applications that include industrial and medical sectors, leading to higher exposure, which in turn increases the concern about their safety. As the toxicity of nanomaterials is still controversial, more high quality  research is needed in order to clarify this matter. The aim of this study was to evaluate the OECD country’s research productivity on nanotoxicity research. For this, SciVerse Scopus was used to search and analyse the documents with specific key words related to nanotoxicology. Our analyses show that 44659 research articles were published worldwide, more than half in the last five years. About 60% of these papers originate from the OECD countries and among these 26590 articles produced by OECD countries about 72% were original articles and 14% were reviews. About 2% of the research articles were published by the Biomaterials Journal (Impact factor 4.2). The most productive country was USA (n =10812 article) with the highest quality of nanotoxicity research (h-index= 82). The significant relation between the countries’ collaboration and quantity – quality of the nanotoxicity researches was observed. The current study emphasizes the need for collaboration in this field between the OECD countries and other countries, especially China and India, which could affect not only the number of the articles but also the quality of the research.

Keywords: bibliometric, OECD countries, nanotoxicity, research productivity

The Distinct Properties of Natural and GM Cry Insecticidal Proteins

Latham1 J.R.; Love2 M.; & Hilbeck3 A.

1The Bioscience Resource Project, Ithaca, NY, USA: email; 2Independent scholar; Melbourne, Victoria, Australia; 3Swiss Federal Institute of Technology (ETH), Zurich, Switzerland.


The Cry toxins are crystal-forming proteins produced by the bacterium Bacillus thuringiensis. There are many distinct Cry toxins which, in their natural forms, vary widely in toxicity towards a very wide range of organisms, from gastropods and nematodes to insects, including Lepidoptera. Cry toxins, such as Cry1, Cry2, and Cry9 are used worldwide in GMO crops to protect against insect pests. It is widely assumed that Cry toxins naturally active against coleopterans will remain active against coleopterans when produced in GMO plants. Some published data contradicts this assumption, showing that Cry toxins produced in GMO plants can have an altered and broader spectrum of activity than naturally-occurring Cry proteins. Such anomalies caused us to catalogue the differences between natural Cry toxins and commercial GMO Cry toxins in a search for an explanation. Our results, which were obtained primarily from documents submitted to support GMO approvals, show that there are potentially important chemical differences between natural and GMO Cry toxins. Natural Cry proteins are crystals, for example, whereas GMO Cry proteins are soluble. GMO Cry proteins are often truncated or mutated by developers or altered by the plant itself. Such differences generally predict a broadened toxicological activity spectrum for the Cry proteins in GM crops because they represent a progression towards the processed and activated form of the protein. This enhancement of toxicity is supported by toxicological studies carried out by applicants, suggesting again that in many cases GMO Cry toxins have greater and broader toxicological activity than natural ones.

Keywords: Cry toxins, Bacillus thuringiensis toxins, risk assessment, GMO, plant


Investigation of the Cytotoxicity of Medical Devices on Cell Cultures of Laboratory Animals

Maryna Anisovich, Nastassia Ahamava, Svetlana Petrova

Republican unitary enterprise «Scientific practical centre of hygiene»



A study of the cytotoxic properties of medical devices was made on cell cultures of various genesis (experiments were carried out on primary cultures of cells of those tissues with which medical devices will interact when used).

The study was conducted in accordance with the recommendations of ISO 10993-5:2011. Extracts of medical devices were studied. As a positive control a solution of ethyl alcohol (1%) was used.

A study of the cytotoxicity of heart implants was conducted on a primary culture of rat cardiomyocyte cells. Suture surgical material was studied on mouse primary dermal fibroblasts. The absence of the cytotoxic effect of the medical devices in vitro was shown.

A scheme for analyzing the cytotoxic effect of medical devices on cell cultures was developed. At the first stage, the cytotoxicity of medical devices was evaluated in rapid tests – in the MTT test (the analysis of metabolic activity) and in the LDH test (the analysis of integrity of cell membranes). At the second stage, if more detailed studies are needed, within 14 days the analysis of the cell population by different methods was carried out. Estimation of growth dynamics of cells, the analysis of cell morphology is conducted microscopically  on 1,3,5,7,11 and 14 day of the experiment. As well the cell population was analyzed by cytofluorimetric methods: determination of the number of apoptotic cells, micronuclei, cell ratio at different stages of the cell cycle.

Keywords: cytotoxic properties, primary cell cultures, MTT test, LDH test, cytofluorimetric methods


Impact of Pesticide Residues in Feed on Animal Health


Anne Schmitt, Katrin Franke and Lars Niemann

German Federal Institute of Risk Assessment (BfR), Berlin


Current legislation on authorization of plant protection and biocidal products requires risk assessment with regard to animal health. At present, no harmonized procedure exists at EU level.

A case study was performed. In cooperation with EFSA (European Food Safety Authority), Germany considered the impact of glyphosate residues in feed on animal health. Relevant information was taken from the Renewal procedure for glyphosate (RAR, 2015). Furthermore, results of a cooperation project (between the BfR and the University of Veterinary Medicine, Hannover) were taken into account. This research project examined the effects of a glyphosate-based herbicidal product on bacterial communities in an in vitro system by means of the ”Rumen Simulation Technique (RUSITEC)”. The relevant toxicological data were compared to the expected exposure of farm animals. For porcine, equine and avian species, no health concerns were found. No final conclusion could be drawn for the assessment of the impact of glyphosate on bacterial communities in the ruminant gastrointestinal tract.

As shown for glyphosate, the database regarding effects on animal health is usually scarce. All possible sources of information have to be considered. Further research is needed, in order to elucidate whether the reference doses established for humans as a general rule can be applied to farm animals or whether different uncertainty factors or critical toxicological effects need to be considered when extrapolating from toxicological studies performed in laboratory animals.

Keywords: risk assessment, farm animals, plant protection products



Acute Deliberate Organophosphate (Coumaphos) Poisoning with Intermediate Syndrome in a One-Year-Old Child


Kiat, WK,

Toxicology Unit, St. Luke’s Medical Center, Quezon City, Philippines


Objective: To report a case of acute organophosphate poisoning in a one-year-old child and development of intermediate syndrome in just eight hours after exposure. Case Report: A case of a previously well one year old child who came in the emergency room because of sudden onset of difficulty of breathing, cyanosis, excessive oral secretions, one episode of diarrhea and weakness noted three hours earlier after ingesting allegedly contaminated powdered milk formula. Vital signs showed a blood pressure of 90/60, cardiac rate of 112 per minute, respiratory rate of 12 cycles per minute and axillary temperature of 35.4 degrees Celsius. ABG analysis showed respiratory acidosis. Endotracheal intubation was immediately done and was hooked on mechanical ventilation. The toxidrome of the patient is compatible with acute cholinergic excess; hence a trial dose of atropine was given with some improvements. Eight hours later, neurological examination showed absence of deep tendon reflexes, no spontaneous respiration, no response to pain, flaccid muscle tone, no neck rigidity and lateralizing signs but with spontaneous eye opening. RBC cholinesterase determination showed a result of 0.057 delta pH/hr, which is significantly depressed. Atropine was given at 0.02 mg/kg intravenously until full atropinization was achieved. Packed RBC transfusion was given. Twenty-four hours later, patient was noted to have response to painful stimuli and spontaneous respiration. A repeat RBC cholinesterase determination showed a result of 0.25 delta pH/hr. Atropine was continued as needed. Test was done on the allegedly contaminated milk using GC-MS, it is positive for Coumaphos. The patient was discharge after seven days.

Conclusion: Intermediate syndrome usually develops within 48-96 hours after acute cholinergic crisis due to prolonged inhibition of cholinesterases.

Keywords: organophosphate, intermediate syndrome, very young child

Pesticide Mixtures Induced Hepato-renal Dysfunction and Oxidative Stress in Pregnant Mice and Their Pups: The Role of Antioxidants


Sameeh Mansour and Marwa Gad

Environmental Toxicology Research Unit (ETRU), Pesticide Chemistry Department, National Research Centre, Dokki, Giza, Cairo, Egypt


In real life, humans are exposed to a cocktail of pesticide residues in food. Women may expose to these pesticide residues during pregnancy. Highly lipophilic compounds, such as organic pesticides, have the ability to cross the placenta and reach the fetus. This study evaluates toxicity of a mixture of three pesticides (atrazine, chlorpyrifos and endosulfan; ACE) added to the rodent diet allowing the mice to ingest the equivalent of the Acceptable Daily Intake (ADI) of each pesticide, in addition to oral administration of vitamin E (α-tocopherol; 100 ul/ mouse). During gestation (21d), the mouse dams were received one of the following treatments: (a) diet free of ACE; (b) diet enriched with ACE; (c) diet free of ACE + oral vitamin E; and (d) diet enriched with ACE + oral vitamin E. During lactation, the dams didn’t receive any chemical treatments. After weaning (42d), selected organs and blood samples were collected for analyses. Compared with the control results either in dams or their pups, the ACE mixture induced high elevation in AST, ALT, ALP, urea and MDA and high decline in BuChE, SOD and CAT. The pups were more affected than the dams with respect to alterations in MDA and BuChE activities, while the opposite was achieved with respect to SOD activities. Supplementation of vitamin E in conjunction with the pesticide mixture revealed the powerful effect of this vitamin; based on the estimated “Amelioration Indices; AI”. The findings revealed the ameliorative effect of vitamin E against toxicity of the tested mixture.

Keywords: pesticide mixture, oxidative stress, vitamin E, amelioration, pregnant mice