جستجوی مقالات مرتبط با کلیدواژه "مزوبوتوس اپئوس" در نشریات گروه "دامپزشکی"

Ten Scorpion samples Mesobuthus eupeus were collected from Baghmalek region in the Khuzestan province of Iran before were identified by Razi Vaccine and Serum Research Institute reference laboratory of Ahvaz. Then, DNA was extracted by phenol/chloroform method in the Laboratory of Molecular Biology in the Faculty of Veterinary Medicine of Shahid Chamran University of Ahvaz. The molecular phylogenetic analysis of Mesobuthus eupeus is carried out based on sequence data of 623 nucleotides fragment of cytochrome C oxidase subunit I. The gene fragments were amplified by PCR using the specific forward and reverse primers. PCR products were fractionated by agarose gel electrophoresis prior to purifying using gel extraction kit. The purified DNA was sequenced by an Applied Biosystems DNA sequencer via Gene Fanavaran Company. In order to confirm the sequencing data, each gene fragment was sequenced in both directions. In order to compare the sequence data with the similar sequences from other scorpions, the target sequence data from different scorpions were retrieved from the Genbank using nblast program via NCBI website. Multiple alignments of the deduced amino acid sequence of cytochrome C oxidase subunit I exhibited 92 and 91% identity to the homologous M. martensii and M. gibossos, respectively. The highest level of identity was scored with M. eupeus philipsi (93%). The results of phylogenetic analysis using cytochrome oxidase subunit 1 indicate that the sequence data of Khuzestan scorpion Mesobuthus eupeus is slightly different from M. eupeusphilipsi gene. As regards of this discrepancy, it concluded that these two Mesobuthus species with highly similar morphological features possibly belonging to two different subspecies.

Considering the lack of knowledge of the molecular biology of scorpion venom antibacterial peptides، as a first step we sequenced and identified a full-length sequence from the venom glands of scorpion (Mesobuthus eupeus) through RT-PCR. Scorpion toxin consists of different types of toxins and enzymes which are encoded by individual genes. Antimicrobial peptides are cytolytic peptides that can rapidly kill a broad range of microbes and have additional activities that impact on the quality and effectiveness of innate responses and inflammation. These peptides had been found from a wide variety of organisms in the last few years. Scorpions were provided and identified by Razi Institute in Ahvaz. The origin of the total RNA was the telson، the last segment of the tail containing the two venom glands. Using RNX plus solution (CinnaGen، Iran)، the total RNA was extracted according to the manufacturer’s instructions. Then، cDNA was synthesized with the extracted total RNA as template and oligo (dT) as primer. In order to amplify cDNA encoding neurotoxin، RT-PCR was performed. The amplified cDNA fragments were sequenced in both strands according to the dideoxy termination method. Sequence comparisons with GenBank database were done using the BLAST software. Based on the sequence data، the complete sequence contained 273 nucleotides called MeBTXA signal peptide sequence was identified، and the glycine at position 20 was assumed to represent the start of the mature protein. Remarkably، they share 96، 94 and 67% similarity with the long-chain potassium ion channel blocker from Mesobuthus martensii، Buthus occitanus Israelis and Tityus costatus. Domain analysis of the protein showed «Arthropod defensin» between amino acid resides 55 to 85. This domain which is rich in cysteine is belongs to the antibacterial peptide family and has effect on the gram positive bacteria. This high sequence similarity together with completely conserved in the positions of all 6 cysteine indicated that this gene is a new member of β-toxin from the Iranian M. eupeus which may originate from a common ancestor.

The venoms of Buthidae scorpions are known to contain basic, single-chain protein -toxins consisting of 60-70 amino acid residues that are tightly cross-linked by four disulfide bridges. Total RNA was extracted from the venom glands of scorpion Mesobuthus eupeus collected from the Khuzestan province of Iran and then cDNA was synthesized with the modified oligo (dT) primer and extracted total RNA as template. By applying the Semi-nested RT-PCR using homologous primers, the fragments of 273 bp was amplified and sequenced. The coding region encodes a polypeptide of 85 amino acid residues with a calculated molecular weight of 9. 337 kDa and theoretical isoelectric point of 6. 52. It was designated as MeNaTx-4. This peptide with homology to the «Scorpion long chain toxin-like domain» is belongs to the Toxin-3 superfamily. Multiple alignment of the putative amino acid sequence of MeNaTx-4 exhibited 95% sequence identity with lesser Asian scorpion M. eupeus venom sodium channel -toxin-3 but interestingly displayed only 63% identity with other -toxins from this species. The sequence was also identical to the counterpart sequences from M. martensii (92%) and Buthus occitanus israelis (80%). This high sequence similarity together with completely conserved in the positions of all 8 Cys indicated that MeNaTx-4 is a new member of -toxin from the Iranian M. eupeus which may originate from a common ancestor.

In tropical and subtropical countries, envenomation by scorpions (so-called scorpionism) represents a serious public health problem. In the present study, the toxic effects of mice LD50 injections of Mesobuthus eupeus (Me) venom on the kidney and liver of anesthetized rabbits were investigated. Six rabbits were selected and ALT, AST, BUN and creatinine were measured at 0, 1 and 3 hours after envenomation and histopathological studies were carried out postmortem. All the animals showed signs and symptoms of envenomation within 30-40 minutes and died 3 to 3.5 hours after venom injection. Histopathological examinations revealed glumerolar congestion, dilated vessels of interstitium and focal interstitial congestion in the kidney and focal hemorrhage, central vein congestion, congested vessels in portal areas and dilated sinusoids in the liver at 3 to 3.5 hrs following venom injection. In addition, biochemical analyses indicated significant rise in the levels of ALT and creatinine following Mesobuthus eupeus envenomation in animals at 3 hrs. However no significant changes were observed at 1 hr. In conclusion, scorpion (Mesobuthus eupeus) venom leads to damage in vital organs such as liver and kidney.

Rheumatoid arthritis is an autoimmune disease that causes chronic inflammation of the joints as well as other organs in the body. Adjuvant-induced arthritis models in inbred rats serve as relevant models for RA, having many clinical similarities to this disease. Using honey bee venom as a treatment for Rheumatoid arthritis is an ancient therapy in various parts of the world. However scorpion venom neurotoxins are responsible for toxicity and pharmacological effects. Twenty-five Wistar male rats weighing 110-130 g were divided in 5 groups and Arthritis was induced in them, using Freund’s adjuvant, except in group 1. In group 2 after the induction of arthritis no treatment was given. Group 3 received Betamethasone as an anti-inflammatory medicine. Venom (5µg/rat) was used in group 4 as a treatment and Group 5 received crude venom (10µg/rat) as treatment, after R.A induction, all the animals received treatment near the site of tibio-tarsal joint subcutaneously. The clinical features of the adjuvant induced arthritis like difficulty in movement and edema in joint appeared 3 days after inoculation of adjuvant. The onset of inflammation was explosive occurring 13-15 days post inoculation with a peak onset at day15. After the treatment of rats, there was a significant reduction in score of arthritis index in all treated animals. The changes in size of tibio-tarsal joint region in groups 4 and 5 which received crude scorpion venom and group 3 with Betamethasone treatment after arthritis development decreased. At the end of experiment, blood collection for WBCs count was carried out. In 2 (untreated rats) and Betamethasone treated rats, there was a significant rise in WBCs count. However in venom treated rats the rise in WBCs was not significant as compared to group 1 rats. The present study demonstrated that the scorpion (Mesobuthus eupeus) venom could be effective as anti-arthritis agent in animal model of acute inflammation. More studies are needed to be carried out to find the mechanism of the venom and exact therapeutic doses of the venom for acting as anti-arthritis agent.