Androctonus Crassicauda Research Articles

Scorpions of the State of Kuwait

This is the first preliminary study on the scorpions of Kuwait. Three species of scorpions were identified, described and illustrated from the State of Kuwait. Identified species belong to family Buthidae: Apistobuthus susanae, Androctonus crassicauda and Compsobuthus pallidus. Measurements were given for Apistobuthus susanae. Morphometric measurements were given for the three species, along with comparison of Apistobuthus susanae and Apistobuthus pterygocercus.

Whole transcriptome sequencing reveals the activity of the PLA2 family members in Androctonus crassicauda (Scorpionida: Buthidae) venom gland.

Phospholipase A2 is the most abundant venom gland enzyme, whose activity leads to the activation of the inflammatory response by accumulating lipid mediators. This study aimed to identify, classify, and investigate the properties of venom PLA2 isoforms. Then, the present findings were confirmed by chemically measuring the activity of PLA2. The sequences representing PLA2 annotation were extracted from the Androctonus crassicauda transcriptome dataset using BLAS searches against the local PLA2 database. We found several cDNA sequences of PLA2 classified and named by conducting multiple searches as platelet-activating factor acetylhydrolases, calcium-dependent PLA2s, calcium-independent PLA2s, and secreted PLA2s. The largest and smallest isoforms of these proteins range between approximately 70.34 kDa (iPLA2) and 17.75 kDa (cPLA2). Among sPLA2 isoforms, sPLA2GXIIA and sPLA2G3 with ORF encoding 169 and 299 amino acids are the smallest and largest secreted PLA2, respectively. These results collectively suggested that A. crassicauda venom has PLA2 activity, and the members of this protein family may have important biological roles in lipid metabolism. This study also revealed the interaction between members of PLA2s in the PPI network. The results of this study would greatly help with the classification, evolutionary relationships, and interactions between PLA2 family proteins in the gene network.

Diet of Pharaoh Eagle-Owl, Bubo ascalaphus, from Ara’r region, northern Saudi Arabia

Abstract The diet of the Pharaoh Eagle-Owl, Bubo ascalaphus, was investigated based on 338 pellets collected from caves and underground caves in Ara’r region, northern Saudi Arabia. Small mammals constituted the highest number of consumed prey (75.75%), followed by arthropods (20%), birds (2.9%) and reptiles (1.26%). The Libyan Jird, Meriones libycus, was the most consumed rodent (26.46%) followed by Sundevall’s Jird, Meriones crassus (20.47%), while the least were Cheesman Gerbil, Gerbillus cheesmani, and Wagner’s Gerbil, Gerbillus dasyurus. At least three species of scorpions, Androctonus crassicauda, Compsbuthus sp. and Scorpio sp., and two species of reptiles (Ptyodactylus hasselquistii and Trapellus agnetae) were recovered. Study of owl pellet contents proved to be a valuable tool to study species composition in unexplored regions. Also, our findings substantiate the fact that the Pharaoh Eagle-Owl is an opportunistic species that adapts to available preys in its habitat.

Antihelimentic effect of Androctonus crassicauda scorpion venom against Trichuris arvicolae isolated from Psammomys obesus in Egypt

Trichuridae family has a genetic and morphological variability between species affecting rodents, but it is considerably hard to morphologically diagnose species within the genus of Trichuris and the individuals of these species are identified according to their host, as it is known that Trichuris spp. is strictly host-specific. However, some species lack host specificity. So, it is necessary to use molecular data in order to well identify the Trichuris spp. in Egyptian rodents. The host examined in the current research is Psammomys obesus and the molecularly identified species from its cecum is Trichuris arvicolae. In addition, Trichuris arvicolae was subjected to in vitro treatment with Androctonus crassicauda Crude Venom as a model of natural alternative treatment for gastrointestinal nematodes that increasingly develop anthelmintic drug resistance. The changes in Trichuris arvicolae were monitored using scanning electron microscopy, Androctonus crassicauda Crude Venom made a significant ultrastructural surface changes in Trichuris arvicolae, including marked cuticular sloughing, disintegrated bacillary glands, bursting of vulva and edema of anal region. This study was done for closer identification of Trichuris spp. infecting rodents in Egypt and evaluating the efficacy of Androctonus crassicauda Crude Venom in vitro.

Molecular Survey of Mitochondrial Genes in Different Populations of the Black Fat-Tailed Scorpion, Androctonus crassicauda.

Androctonus crassicauda is the most medically relevant scorpion and understanding its genetic forms is essential for improvement of anti-venom sera, and risk management of scorpionism. Present study was designed to identify the variations of mitochondrial genes in different populations of A. crassicauda. Adults of A. crassicauda were collected from Zanjan Province during 2016-2017. Genomic DNA of samples was extracted and fragments of mitochondrial 16S, COI and ND1 genes were amplified and some of the amplicons were sequenced. Haplotype of samples were identified by multiple alignment of sequences, then phylogenetic trees of haplotypes were constructed. Fragments of 352bp, 618bp and 680bp were amplified from 16S, COI and ND1 genes respectively. Nucleotide sequence in COI fragments was conserved, however, five haplotypes with some specific polymorphic sites were detected in 16S and ND1 fragments. Haplotype I was dominant and found in all areas. Other haplotypes were rare and limited to specific regions. Analysis of the phylogenetic trees inferred from 16S and COI genes, confirmed a strong positive correlation between geographic and genetic distance. Mitochondrial COI, 16S and ND1 genes were detected suitable for identifying the population structure. Five genotypes were found using 16S and ND1 genes. To prepare and improve the anti-venoms quality, additional studies are necessary to identify the toxin electrophoretic profile and geographical/ecological niche models of these genotypes in future.

Şanlıurfa İlinde Yayılış Gösteren Androctonus crassicauda:Buthidae Türünün Biyo-Ekolojisi Üzerine Araştırmalar

Androctonus crassicauda scorpion species lives in nature both as a prey and a predator. So it has venom that can be effective for hunting and protection. It can cause venoming and death by stinging people and animals. It is of great importance to know the ecological characteristics and density of this scorpion species, especially in regions where venoming cases are high. In the present study, Şanlıurfa province, where the scorpion species Androctonus crassicauda is prevalent, was chosen as the research area. 289 samples were collected after field studies in all districts of Şanlıurfa province. Owing to desolated and stony structure, Androctonus crassicauda was observed to be more intensive in Harran district. It was generally found from April to October. The most abundant period of the species is June, July, and August. Considering the seasonal expectation, test results of correlation for a series with non-normal distribution were listed in two options, Kendall'stau_b correlation coefficient and Spearman'srho correlation coefficient. There was a positive correlation of 0,168 (16%) at 1% significance between month and population for the former (Kendall'stau_b) coefficient. A positive correlation of 0,231 (23%) at 1% significance level between month and population for the latter (Spearman'srho) coefficient. In the view of regional expectation, results of correlation test for a non-normal distribution were presented in two options. A negative correlation of 0,099 (9%) was found at 5% significance level between region and population for the former coefficient. There was a negative correlation of 0,128 (12%) at 5% significance level between region and population for the second coefficient. The study revealed a variation between seasons and districts.

Discrimination of two species (Androctonus crassicauda and Leiurus abdullahbayrami; Buthidae Scorpions) by MALDI-TOF- MS-based PCA

Teh venoms of teh scorpions Androctonus crassicauda and Leuris abdullahbayrami, scorpion species each of teh two members of teh Buthidae family, were analyzed by MALDI–TOFMS in a mass range between 1 and 50 kDa. Interspecies differentiation was evaluated over peptide and protein molecules in dis mass range. Teh similarities and differences between two different scorpion species were revealed wif teh TEMPprincipal component analysis study, which was conducted wif spectral patterns including peptide and protein profiles. Teh similarity rate of teh LAB-123 and teh LAB460 scorpion venoms of teh same species was found as 66% while teh similarity rates of venoms of teh ACR species to teh LAB species ranged from zero to 37%. It was demonstrated dat scorpion venoms belonging to two different species from teh Buthadia family can be differentiated wif teh halp of dendrogram and gel profile, CCI color matrix, 3D or 2D-scattering profile, spectral mass loading data formed by peptide and protein spectral patterns of eleven scorpion venoms. It is anticipated dat dis approach, which was used for teh first time wif teh application of MALDI-TOFMS-based PCA analysis for teh differentiation of scorpion venoms, will be useful in differentiating venoms wif different spectral patterns.

Morphometric Indices and Venom Protein Profile in Different Populations of Androctonus crassicauda.

Androctonus crassicauda is the most medically relevant animal and understanding its morphological characteristics is essential in the production of antiscorpion sera. Adults of A. crassicauda were collected from different areas of Zanjan Province and the morphometric parameters and the cuticular fluorescence patterns of samples were studied. The crude venom of samples was extracted by electric stimulation, and their biochemical properties were analyzed by the SDS-PAGE method. Values of the morphometric parameters depended on sex and altitude of the area. Except for values of the pectinal organ, these parameters in females were higher than in males. No significant difference was in the number, shape, and intensity of cuticular fluorescence patterns. The body length of males in high and lowlands was 72.53±1.53 and 77.33±2.70mm, respectively. Females' body lengths in that area were 81.66±2.19 and 86.55±2.33mm, respectively. Analysis of toxin proteins showed two isotypes that the 12, 13, 15, 16, 18, and 19kDa proteins were in all areas. However, the 41 and 74kDa proteins, and 46 and 63kDa proteins were detected in low and highlands, respectively. Black fat-tailed scorpion has a considerable dominancy and developing preventive programs and providing treatment facilities in studied areas are necessary. Values of the morphological parameters and venom electrophoresis patterns depended on the geographical location. Therefore, pool crude toxin is suggested for the production of effective antivenoms. Moreover, additional field complementary works in the geographic information system based niche modeling and mass fingerprinting of scorpion venoms are suggested for screening effective isotypes.

Morphometric and meristic diversity of the species Androctonus crassicauda (Olivier, 1807) (Scorpiones: Buthidae) in Saudi Arabia.

Intraspecific molecular and morphological variations among geographically isolated populations are useful for understanding the evolutionary processes, which is considered early stage of allopatric speciation. Also, the knowledge of the regional variation of scorpion venom composition is needed to improve antivenom therapeutic management. Androctonus crassicauda (Olivier, 1807) is the most common and medically important species in Arabia and the Middle East. Therefore, this study aimed to investigate the geographic morphological variation among A.crassicaudapopulations, regarding its geographical distribution in unexplored arid regions in Saudi Arabia. Samples were collected and examined morphologically under a dissecting microscope from different four eco-geographical regions. The results of ANOVA and multivariate statistical analyses provide strong evidence of geographical variation. The two populations from OTU3 and OUT4 showed the greatest degree of morphological difference from populations of OUT1 and OUT2. Each OTU3 and OTU4 populations showed significant speciation without overlapping in the two groups, while the remaining overlapped groups comprised two other populations. Several body variables influenced male separation, including carapace posterior width, metasoma 3rd length, and metasoma 2nd length. For females, telson length, metasoma 1st width, and sternite 7th width were highly influential variables. Such variation may suggest the existence of cryptic taxa within A. crassicauda populations in Saudi Arabia. Moreover, metasoma ratios can be used as good indicators in intraspecific variation studies of Scorpions.

Preparation and radiolabeling of Iranian Androctonus crassicauda scorpion venom with technetium-99m as a new agent for cancer diagnostic and therapeutic purposes

Abstract One of the innovative methods in cancer treatment with fewer side effects is the use of active principles produced by animals and microorganisms to generate new drugs with diagnosis and treatment capability for cancer. Androctonus crassicauda scorpion venom has high potential as an anti-tumor agent and has the ability to interfere with human physiology but is still unknown. Therefore, labeling venom with radionuclides is very important to study the biological distribution of toxins for diagnostic and therapeutic purposes of various diseases, especially cancer. In this research, the toxic fraction of this venom was labeled with 99mTc. The radiochemical purity of the labeled toxic fraction was obtained by using chromatographic systems. Animal biodistribution studies were performed after injecting the labeled compound into normal rats and 4T1 breast cancer-bearing mice. The radiochemical purity of 90% was obtained for labeled compound. Biological distribution studies showed relatively rapid clearance of the labeled compound from the blood circulation system. The accumulation of the labeled compound in the liver and kidney was the highest among the organs, which could indicate the possibility of toxin excretion metabolism from liver and the kidneys. The accumulation rate of the labeled compound after 15 min was 7% in the breast cancer mass, and the ratio of the distribution of the labeled compound in the breast cancer mass to the blood was 155% at 15 min post injection. The study results indicated that scorpion venom labeling with 99mTc could be a useful tool for the biodistribution and kinetic studies of the venoms for clinical use. On the other hand, based on the results of the biological distribution of the labeled compound in tumor-bearing mice, the 99mTc-venom complex can be used as an imaging agent to diagnose breast cancer. Moreover at the same time, with some measures, it can be used as a therapeutic agent for breast cancer with more consideration. We hope that the results of this study will be a step in the future for the clinical diagnostic and therapeutic purposes of cancer using natural products.

Differential venom gland gene expression analysis of juvenile and adult scorpions Androctonus crassicauda

BackgroundThe Androctonus crassicauda, belonging to the genus Androctonus of the family Buthidae, is the most venomous scorpion in Middle East countries. However, the venom gland transcriptome profile of A. crassicauda scorpion has not yet been studied. In this study, we elucidated and compared the venom gland gene expression profiles of adult and juvenile male scorpion A. crassicauda using high-throughput transcriptome sequencing. This is the first report of transcriptional analysis of the venom glands of scorpions in different growth stages, with insights into the identification of the key genes during venom gland development.ResultsA total of 209,951 mRNA transcripts were identified from total RNA-seq data, of which 963 transcripts were differentially expressed (DE) in adult and juvenile scorpions (p < 0.01). Overall, we identified 558 up-regulated and 405 down-regulated transcripts in the adult compared to the juvenile scorpions, of which 397 and 269 unique unigenes were annotated, respectively. GO and KEGG enrichment analyses indicated that the metabolic, thermogenesis, cytoskeleton, estrogen signaling, GnRH signaling, growth hormone signaling, and melanogenesis pathways were affected by two different growth conditions and the results suggested that the DE genes related to those pathways are important genes associated with scorpion venom gland development, in which they may be important in future studies, including Chs, Elovl, MYH, RDX, ACTN, VCL, PIP5K, PP1C, FGFR, GNAS, EGFR, CREB, CoA, PLCB, CALM, CACNA, PKA and CAMK genes.ConclusionsThese findings broadened our knowledge of the differences between adult and juvenile scorpion venom and opened new perspectives on the application of comparative transcriptome analysis to identify the special key genes.

Identification, characterization, and molecular phylogeny of scorpion enolase (Androctonus crassicauda and Hemiscorpius lepturus)

Scorpions are the oldest terrestrial fauna with an ancient evolutionary history. To study the origins of scorpions, we conducted a molecular phylogenetic study of the arthropod using slowly evolving enolase sequences. In this study, we used deep RNA-seq technology to identify the enolase sequences of Androctonus crassicauda and Hemiscorpius lepturus scorpions. The scorpion’s enolase sequences were in silico analyzed to predict the protein structures, physicochemical characterization, and exon-intron organization. The enolase of the scorpion was found to be a protein of 433 amino acids with a molecular mass of ∼47 kDa and an acidic isoelectric point. Here, we reported the existence of 8 exons and an inner GC/AG intron for scorpion enolase. Using homology modeling, the enolase of the scorpions was found to be very conservative during the evolution process. Furthermore, our phylogenetic analysis indicated the independent evolution of the Buthidae family from other scorpions and also the sister group relationship of Hemiscorpionidae with the Caraboctonidae families. The scorpions’ enolase sequences demonstrated a close fit to those of spiders, followed by mite counterparts. This strongly supports our phylogeny analysis, which indicates a sister-group relationship between scorpions and spiders and their common ancestors with mites. This study provided support for the pairing of scorpions with spiders, mites, etc. using enolase as a nuclear gene. Our molecular clock analyzes of the arthropod show that the most recent common ancestor of arachnids and insects were diversified in the Cryogenian period of Neoproterozoic era, earlier than dates reported from fossil records.

Intraspecific molecular variation among Androctonus crassicauda (Olivier, 1807) populations collected from different regions in saudi arabia

Androctonus crassicauda is one of the medically important scorpion species occurring in the Middle East. In this study, molecular variation in mtDNA cytochrome oxidase subunit I (COI) gene within seven populations of A. crassicauda from four main ecological regions of Saudi Arabia was investigated. Scorpion specimens were collected from different eco-geographical regions. DNA was extracted, subsequently 700 bp fragments of COI gene were amplified using specific scorpion primers and sequenced. The obtained sequences were analyzed, and phylogenetic trees were constructed. The phylogenetic analysis showed high levels of genetic variation among A. crassicauda populations with the existence of three distinct lineages. Also, it supports the existence of two distinct populations of A. crassicauda in Saudi Arabia, which perhaps suggestive of a putative distinct species. Further morphological studies with additional specimens from the Arabian Peninsula may reveal possible undiscovered and cryptic species in the region.

Genetic diversity and population structure of Androctonus crassicauda (Scorpiones: Buthidae) in different ecogeographical regions of Saudi Arabia and Iran

The genetic structure of four populations of Androctonus crassicauda (Olivier, 1807) from different ecogeographical regions of Saudi Arabia was studied. To detect genetic differences among populations, the sequences of the cytochrome oxidase subunit I gene of 21 individuals of A. crassicauda from Saudi Arabia, and 11 sequences from Iran were utilised. The nucleotide and haplotype diversity were constructed. The results revealed a high level of haplotype and nucleotide diversity within all populations. A total of eighteen haplotypes were identified, 17 of which were unique. Molecular variance analysis (AMOVA) indicated a high genetic diversity reflecting differences between the populations. Fixation indices revealed significant genetic structure among all populations from Saudi Arabia and Iran. Also, the phylogenetic tree and the haplotype network analyses show the existence of three distinct clusters of A. crassicauda in Saudi Arabia and Iran.

Scorpion crude venom induced apoptosis and structural changes of Echinococcus granulosus protoscolices

Echinococcus granulosus parasitic infection in humans causes hydatidosis (also known as echinococcosis), which is a severe zoonotic illness that puts public health at risk.This research was designed to assess the status of morphological, ultrastructure, and cell apoptosis of E. granulosus protoscolices. For this purpose, the protoscolices were isolated and collected from the hydatid cysts in the lungs and livers of camels, then incubated for 4 h in Androctonus crassicauda crude venom (ACCV) (at the concentrations of 50 and 100 µg/ml). The changes of E. granulosus protoscolices were monitored using light, transmission, and scanning electron microscopy, and the caspase-3 expression was detected by immunohistochemistry. The current study results have demonstrated the caspase-3 expression was substantially higher in the protoscolices incubated at 100 µg/ml than 50 µg/ml. Also, ACCV caused distinct changes in the morphology and ultrastructure of protoscolices detected by light, TEM, and SEM microscopy. In conclusion, Androctonus crassicauda crude venom (ACCV) may provide an alternative and nonsurgical treatment for hydatidosis. Hence, further studies are recommended to explore the impact of ACCV on inducing apoptosis and structural changes for this parasitic disease.

Recombinant expression and antigenicity of two peptide families of neurotoxins from Androctonus sp.

Scorpion neurotoxins such as those that modify the mammalian voltage-gated sodium ion channels (Nav) are the main responsible for scorpion envenomation. Their neutralization is crucial in the production of antivenoms against scorpion stings. In the present study, two in silico designed genes - one that codes for a native neurotoxin from the venom of the Anatolian scorpion Androctonus crassicauda, named Acra 4 - and another non-native toxin - named consensus scorpion toxin (SccTx) obtained from the alignment of the primary structures of the most toxic neurotoxins from the Middle Eastern and North African scorpions - were recombinantly expressed in E. coli Origami. Following bacterial expression, the two expressed neurotoxins, hereafter named HisrAcra4 and HisrSccTx, were obtained from inclusion bodies. Both recombinant neurotoxins were obtained in multiple Cys-Cys isoforms. After refolding, the active protein fractions were identified with molecular masses of 8,947.6 and 9,989.1 Da for HisrAcra4 and HisrSccTx, respectively, which agreed with their expected theoretical masses. HisrAcra4 and HisrSccTx were used as antigens to immunize two groups of rabbits, to produce either anti-HisrAcra4 or anti-HisrSccTx serum antibodies, which in turn could recognize and neutralize neurotoxins from venoms of scorpion species from the Middle East and North Africa. The antibodies obtained from rabbits neutralized the 3LD50 of Androctonus australis, Leiurus quinquestriatus hebraeus and Buthus occitanus venoms, but they did not neutralize A. crassicauda and A. mauritanicus venoms. In addition, the anti-HisrAcra4 antibodies did not neutralize any of the five scorpion venoms tested. However, an antibody blend of anti-HisrAcra4 and anti-HisrSccTx was able to neutralize A. crassicauda and A. mauritanicus venoms. Two recombinant Nav neurotoxins, from different peptide families, were used as antigens to generate IgGs for neutralizing scorpion venoms of species from the Middle East and North Africa.

In Vitro Efficacy of the Venome of Black Scorpion (Androctonus crassicauda) on Leishmania tropica Promastigotes

Scorpion venom is a substance that shows strong neurotoxic effects with its complex protein content and thus plays an important role for the scorpion in catching and digesting the hunt. Human body stung by a scorpion can show life-threatening systemic effects in a short time, ranging from erythema, pain, edema and local fever to abdominal pain, nausea and vomiting, diplopia and even coma. Scorpion venome is known to possess antimicrobial activity, and some of its compounds have potent antibacterial and antifungal activities. Leishmaniasis is a common vector-borne parasitic infection caused by Leishmania sp. protozoa and can lead skin, mucosa and fatal internal organs involvement in patients . There is a need for new drugs in the treatment of leishmaniasis, because it has been documented lately that there is a growing resistance against antimonial compounds which have been used in its treatment for decades. Leishmania species are known to be susceptible to antimicrobial peptides that act as ion-channel inhibitors, which are known to be present in scorpion venome. In this study, it was aimed to investigate the anti-leishmanial activity of scorpion venome extract obtained from Androctonus crassicauda species in our country. In this context, Leishmania tropica promastigotes which were thawed from liquid nitrogen in our laboratory and first grown in NNN and then RPMI-1640 media, were exposed to different dilutions of the extract containing A.crassicauda venom and meglumine antimonate used in the standard treatment of leishmaniasis and the efficacies on the promastigotes were compared and measured in vitro. This was followed by XTT cell viability test, which assessed whether anti-leishmanial dose of the extract was lethal for human cells or not. Trials showed that the half maximal inhibitory concentration (IC50) values of the venome extract and meglumine antimoniate were 18.12 µg/ml (17.33-18.94) and 8.411 µg/ml (7.922- 8.927), respectively. This preliminary study showed that scorpion venome can be lethal on L.tropica promastigotes in vitro, on relatively higher doses compared to meglumine antimonate. Next step will be to determine the anti-leishmanial proteins in the extract and thus to identify new drug candidates with more specific studies.

Expression and purification of recombinant alpha-toxin AnCra1 from the scorpion Androctonus crassicauda and its functional characterization on mammalian sodium channels.

Alpha-scorpion toxins with long-chain peptide and four disulfide bonds represent diverse pharmacological profiles for various subtypes of voltage-gated sodium channels. Obtaining the natural toxins are difficult and time-consuming process, which represents the major difficulty to interpreting analysis of their structural and functional properties. This study describes the toxin peptide and plasmid construct containing the gene coding for mammalian toxin AnCra1 from the scorpion Androctonus crassicauda venom. We have established genetic construction of fusion protein in pET32a + vector containing thioredoxin (Trx-tag), enterokinase cleavage site and 6xhistidine-tag for efficient expression in Escherichia coli strain RG2 (DE3). The soluble expressed peptide, then purified by Ni-NTA resin affinity chromatography and its purity was confirmed by reverse-phase HPLC and mass spectrometry (7433.54Da.). The electrophysiological data showed that recombinant AnCra1 selectively inhibits the fast inactivation of hNav1.7 channel (EC50 = 136.7 ± 6.6nM). Our findings demonstrate that the AnCra1 is structurally and functionally analogous to alpha excitatory toxins; furthermore, expression and purification of bioactive scorpion toxins in bacterial cells can be a practicable and efficient way to obtain a novel source of toxin peptides as tools to study the function and physiological responses of ion channels.

Toxicity and protein composition of venoms of Hottentotta saulcyi, Hottentotta schach and Androctonus crassicauda, three scorpion species collected in Iran.

BackgroundScorpion stings comprise a serious problem throughout the globe, especially in regions where they are more frequent. Despite a recent upsurge of interest in scorpion venoms by various research groups, there remain many challenges.ObjectiveTherefore, in this study, we aimed to study the toxicity and protein composition of venoms of Hottentotta saulcyi, Hottentotta schach and Androctonus crassicauda, three scorpion species collected in Iran.Materials and MethodsScorpion species were collected from Esfahan farm scorpion company and maintained in the laboratory in containers that mimic their natural habitat. Venom was extracted from A. crassicauda, H. schach and H. saulcyi by electrical stimulation of 8 and 10 V. The toxicity of each venom was established by using four groups of male Swiss albino mice aged 2 months (weighting 18–20 g) for testing each dose of venom. One group was used as a control. Venom was injected into mice by subcutaneous route. Then, animals were monitored for 24 h and LD50 was estimated by the graphic method of Miller and Tainter. Thus, high‐performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method was used to determine amino acids in the venom, and protein concentrations were determined by the Biuret method.ResultsLD50 of scorpion venoms by subcutaneous route was found to be 1.70 mg/kg b.w (A. crassicauda), 1.47 mg/kg b.w (H. saulcyi) and 0.85 mg/kg b.w (H. schach). A. crassicauda, H. saulcyi and H. schach contain 26, 30, and 31 amino acids, respectively. A. crassicauda contains low concentrations of alpha‐aminoadipic acid, beta‐aminoisobutyric acid, beta‐alanine and citrulline. H. saulcyi contains a concentration of hydroxylysine, whereas H. schach has no such concentration. A. crassicauda also had the highest levels of tyrosine and threonine. Only A. crassicauda venom contains a low proportion of proteins (14.80%) compared with those of H. schach (16.26%) and H. saulcyi (16.20%). Albumin content in the venoms was 11.7% (H. saulcyi), 5.4% (H. schach) and 4.4% (A. crassicauda).ConclusionScorpions venoms have a variable toxicity and an interesting composition in amino acids and proteins. Work on the development of anti‐venom is fundamental.

The effects of antivenom administrations on the brain tissue of experimentally envenomed pregnant rats and their pups with Androctonus crassicauda scorpion venom during organogenesis period

This study aims to show the changing effects of Androctonus crassicauda venom and A. crasicauda specific antivenom during pregnancy in brain tissue of dams and their pups. Totally, 12 pregnant-Wistar Albino rats were randomly divided into two groups as venom-antivenom administration (n = 6) and control groups (n = 6). In venom-antivenom administration group (VAV), the sublethal dose of A. crassicauda venom dissolved in 1 mL physiological saline solution was subcutaneously (s.c.) injected into pregnant rats during organogenesis period (between 7 and 13 days of pregnancy). Four hours after each venom injection, 1 mL/s.c. dose of the specific anti-venom was administered to rats of VAV group. The rats in control group were given sterile saline solution 1 mL/s.c. In both groups, the fetuses were surgically delivered on the 21st day of pregnancy; dams and pups were sacrificed on postnatal 21 days, and their brain tissues were removed. The brain tissue of dams and their pups were evaluated histopathologically and immunohistochemically. To show the neuronal damages, 8-hydroxy-2-deoxyguanosine (8-OHDG) and amyloid beta precursor protein (ABPP) immunoexpressions were scored in cerebrum, cerebellum, pons and medulla oblongata of brain. To show the neuroprotection, reelin and beta-arrestin immunoexpressions were scored again in the same way. In this context, 8-OHDG immunoexpressions were increased in neocortex, hippocampus and nucleus accumbens when compared with that of control group. Amyloid beta precursor protein was negative in both groups. Reelin and beta-arrestin partly increased in fore and mid brain of VAV group as a reaction against neuronal damages when compared with that of control pups. The authors believe that prompt intervention using anti-venom to scorpion envenomation can partly stop neuronal damages. This neuroprotection may be increased to high and serial doses of anti-venom to save neonatal lives.