Urinary metabolites of diazinon and chlorpyrifos in sprayer operators and farm workers of a potato farm.

In order to investigate the effect of diazinon and chlorpyrifos on agricultural workers exposed to pesticides, urinary metabolites 2-Isopropyl-6-methyl-4-pyrimidinol (IMPy) and 3,5,6-Trichloro-2-pyridinol (TPCy) in farm workers, sprayer operators, and non-exposed people as a control group were measured. The modified QuEChERS method was applied to extract samples and was measured using a gas chromatograph/nitrogen-phosphorus detector. The obtained results showed that the median concentrations of TCPy were 36.92-547.7 and 7.7-49.58 ng/mL for sprayer operators and farm workers, respectively. Moreover, the median concentrations of IMPy were 81.66-593.1 ng/mL for sprayer operators and 40.6-66.1 ng/mL for farm workers. The control group had no measurable metabolites. The IMPy level of 60% of sprayer operators was significantly higher (P ˂ 0.05) than the TCPy level. The analysis of variance highlighted the significant relationship (P ˂ 0.05) between the levels of each metabolite and the use of safety gloves, respiratory masks, safety goggles, working time per week, and type of insecticide exposure. Our findings revealed the need to measure the urinary metabolites of these insecticides in other exposed workers. Also, workers should be taught the impact of using personal protective equipment.

OdTx12/GNA, a chimeric variant of a ß excitatory toxin from Odontobuthus doriae, reveals oral toxicity towards Locusta migratoria and Tenebrio molitor.

OdTx12, a beta excitatory toxin from Odontobothus doriae had previously been identified and characterized. It had been shown that OdTx12 causes significant lethal effects on insects by injection but does not show any toxicity on mice. Due to the natural ineffectiveness of scorpion toxins to act as oral toxins, OdTx12 was fused to Galanthus nivalis agglutinin (GNA), a protein with the potential to cross the insect gut. The sequence of OdTx12/GNA gene was chemically synthesized, cloned in Escherichia coli, and expressed. The effect of the purified fusion protein (OdTx12/GNA) was assessed on the insect and mammalian cell lines, insect larvae and mice. Toxicity assay on insect cell culture (SF9 cell line) showed comparable toxicity between OdTx12 and OdTx12/GNA (LD50 of 0.0030 and 0.0048 µM, respectively). Also very similar mortality rates were observed by injecting OdTx12 and OdTx12/GNA to Locusta migratoria and Tenebrio molitor. Oral administration of OdTx12/GNA, after five days of feeding, resulted in 96.6% and 98.3% mortality of L. migratoria and T. molitor larvae with an LC50 of 0.69 and 0.43 nmol/g of insect food, respectively, while OdTx12 alone did not cause any toxic effects on the larvae orally, suggesting the role of GNA in delivering the toxin to the insect's haemolymph. No toxicity or mortality was observed after toxicity testing of OdTx12/GNA on a mammalian cell line (HEP-2) or any mortality in vivo, by testing the protein in the laboratory mouse. Herein, we demonstrated that the fusion protein OdTx12/GNA could be considered an effective toxin for the biological control of insects.

The Effect of Different Doses of Mesobuthus eupeus (Scorpionida: Buthidae) Scorpion Venom on the Production of Liver Necrosis in Nmri Mice.

BACKGROUND: Scorpion venom has a variety of different components considerably. Some of these compounds are proteins such as Phospholipase A2 which is one of the most important. Use of scorpion venom for the treatment of any disease requires an initial study to determine the therapeutic dose or safe dose. Therefore, due to the necessity of studying scorpion venom, it is of special importance to study the effects of its dose response in animal tissues. METHODS: To determine the inflammatory effects of scorpion's venom (Mesobuthus eupeus), 50 Nmri mice with an average weight of 24±7g were selected for investigation in two experiments. In first-round 25 of them were divided into 5 groups and were exposed to different doses of venom injection paralleling the control group. Single-injection of various doses on 25 mice was performed and results were compared. RESULTS: There was a significant differences between the test and control groups (in most groups). Liver necrosis was one of the important symptoms in this study, the severity of which was measured and statistically analyzed. CONCLUSION: It was determined that 0.05ppm is a safe dose and sub-lethal doses can use for the investigation of therapeutic effects of venom on cancer, diabetes, dermatitis, and so on.

Identification of Cellulolytic Bacteria from Guts of Microcerotermes diversus Silvestri (Isoptera: Termitidae) in Southern Iran.

Termites are a large and important group of insects in terrestrial ecosystems that decompose lignocelluloses. Among these, Microcerotermes diversus Silvestri (Termitidae) is a destructive invasive pest in many tropical and subtropical regions. In the present study, M. diversus specimens were collected from traps in Ahvaz, Bandar Abbas, Kish, and Khark Islands. Sample suspensions were prepared in 5-ml DH2O and cultured on nutrient agar (NA) medium. All 47 representative bacterial isolates were evaluated for cellulolytic activity by growing them on cellulose Congo-red agar medium. Based on some key phenotypic characteristics, the isolates were tentatively identified at the genus level. These were confirmed by 16S rRNA analysis using a universal primer pair (P1/P6). Sequence alignments revealed that most of the isolates are novel species so far found in the termite guts. Results showed that some of the isolates are common for all surveyed areas. However, there were significant differences in their numbers and degree of cellulolytic activity. The species reported here for the first time for termites of Iran are Bacillus wiedmanii, B. paramycoides, Elizabethkingia anophelis, Lysinibacillus pakestanensis, Pseudomonas hibiscicola, Actinetobacter pitti, A. venetianus, and Ochrobactrum anthropi.

Cloning and expression of OdTx12, a ß excitatory toxin from Odontobuthus doriae, in Escherichia coli and evaluation of its bioactivity in Locusta migratoria.

The venom of Odontobuthus doriae contains several peptide toxins that interfere with the sodium channel function of cell membranes, some of which specifically act on the insect's sodium channel without affecting mammalian cells. In this study sodium channel toxins of Odontobuthus doriae were aligned to other closely related toxins by BLAST and ClustalW servers. Among these toxins, NaTx12 (OdTx12) showed more than 90% similarity to the most known beta excitatory toxin, AaHIT1; furthermore, our modeling studies confirmed high tertiary structure similarity of these proteins. OdTx12 was cloned and expressed in E.coli, using pET26-b and pET28-a expression vectors. Tris-tricine SDS-PAGE and Western blot analysis showed OdTx12 expression by pET28-a, only. After purification, bioactivity of the purified protein was analyzed by injection and oral administration to Locusta migratoria larvae, and toxicity to mammals was tested on mice. Injection of OdTx12 resulted in the killing of larvae with LD50 of 0.4 and 0.2 after 48 and 72 h respectively, but oral administration of OdTx12 had no significant effect on Locusta migratoria, nor did the injection to mice show any signs of toxicity. These results showed that OdTx12, as a novel ß excitatory toxin can be considered as a candidate for insect control purposes.

Sublethal Effects of Some Insecticides on Functional Response of Habrobracon hebetor (Hymneoptera: Braconidae) When Reared on Two Lepidopteran Hosts.

The effects of four commonly used pesticides, diazinon (EC 60%), phosalone (EC 35%), fipronil (Granular formulation 0.2%), and pyriproxifen (EC 10%), on functional response of Habrobracon hebetor (Say) (Hymenoptera: Braconidae) were investigated using two lepidopteran hosts, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) and Heliothis viriplaca (Hufnagel) (Lepidoptera: Noctuidae). Laboratory bioassays determined diazinon and fipronil as highly toxic insecticides for all developmental stages of the parasitoid, while the acute toxicity of phosalone and pyriproxyfen was moderate to very low depending on the life-stage studied. Larval, pupal, and adult stages of the parasitoid were exposed to sublethal concentrations (LC30) of insecticides, and the newly mated females were used to study functional response of the parasitoid to different host densities. With a single exception, a type II functional response was found for control and all insecticide treatments on all life stages and both host species. All insecticides tended to decrease the asymptote of the functional response curve and the maximum parasitism rate, probably by negatively affecting the searching efficiency (a') of the parasitoid. These results indicate that the control efficiency of parasitoids may be negatively affected by sublethal doses of pesticides, even though the type of functional response remains unchanged. As a potential solution, appropriate timing of pesticide application is required to avoid the antagonistic interactions with natural enemies under integrated management programs.

Effect of Iranian Honey bee (Apis mellifera) Venom on Blood Glucose and Insulin in Diabetic Rats.

BACKGROUND: Diabetes is an important disease. This disease is a metabolic disorder characterized by hyperglycemia resulting from perturbation in insulin secretion, insulin action or both. Honey bee venom contains a wide range of polypeptide agents. The principle components of bee venom are mellitin and phospholipase A(2). These components increase insulin secretion from the ß-cells of pancreas. This study was conducted to show the hypoglycemic effect of honey bee venom on alloxan induced diabetic male rats. METHODS: Eighteen adult male rats weighting 200±20 g were placed into 3 randomly groups: control, alloxan monohydrate-induced diabetic rat and treated group that received honey bee venom daily before their nutrition for four months. Forty eight hours after the last injection, blood was collected from their heart, serum was dissented and blood glucose, insulin, triglyceride and total cholesterol were determined. RESULTS: Glucose serum, triglyceride and total cholesterol level in treated group in comparison with diabetic group was significantly decreased (P< 0.01). On the other hand, using bee venom causes increase in insulin serum in comparison with diabetic group (P< 0.05). CONCLUSION: Honeybee venom (apitoxin) can be used as therapeutic option to lower blood glucose and lipids in diabetic rats.

Effect of honey bee venom on lewis rats with experimental allergic encephalomyelitis, a model for multiple sclerosis.

Multiple sclerosis (MS) is a progressive and autoimmune neurodegenerative disease of the central nervous system (CNS). This disease is recognized through symptoms like inflammation, demyelination and the destruction of neurological actions. Experimental allergic encephalomyelitis (EAE) is a widely accepted animal model for MS. EAE is created in animals by injecting the tissue of myelin basic protein (MBP), CNS, or myelin oligodendrocyte glycoprotein (MOG) along with the adjuvant. EAE and MS are similar diseases. Honey Bee venom (Apis mellifera) contains a variety of low and high molecular weight peptides and proteins, including melittin, apamin, adolapin, mast cell degranulating peptide and phospholipase A2. Bee venom (BV) could exert anti-inflammatory and antinociceptive effects on the inflammatory reactions. The guinea pig spinal cord homogenate (GPSCH) is with the Complete Freund's Adjuvant (CFA), consisting of 1 mg/mL Mycobacterium tuberculosis. It was used for inducting EAE in Lewis rats for creating the MS model. The hematoxylin and eosin and luxol fast blue methods were used respectively in analyses of inflammation and detection of demyelination in the central nervous system. Furthermore, the ELISA and the high performance liquid chromatography (HPLC) were used for the assessment of tumor necrosis factor alpha (TNF-α) and nitrate in rats serum. In this study, we indicated that the treatment of EAE with Bee venom decreased the symptoms of clinical disorder, pathological changes, inflammatory cell infiltration, demyelination in the central nervous system, level of serum TNF-α, and the serum nitrates in rat EAE induced through GPSCH.