Identification, classification, and characterization of alpha and beta subunits of LVP1 protein from the venom gland of four Iranian scorpion species.

Scorpion venoms contain bioactive peptides and proteins. Some, can be used for pharmaceutical purposes. So, identification of venom proteins matters because, in addition to determining the function of the toxins can also be an excellent guide to developing new drugs. Here, we got transcriptome of venom glands from four Iranian scorpion species, including Hemsicorpius lepturus, Mesobuthus eupeus, Andructunus crassicuada, and Hottentotta saulcyi using cDNA library synthesis and high-throughput transcriptomic analysis of the venom glands. In a comparative way, we identified the cDNA encoding isoforms of subunits (alpha and beta) of BotLVP1/BmLVP1-like protein in the venom gland of three species except for H. lepturus. Characterization and structure determination of the LVP1_like proteins combined with gene map analysis provided evidence of the existence of some isoforms of LVP1_like proteins, encoded by genes with two exons and one intron, which can be classified in CSαß superfamily in the venom gland of three Iranian scorpion species. According to the high similarity with BotLVP1 and BmLVP1, these proteins could also be potent to mediate cholesterol homeostasis. However, further research is needed to prove it, and this study just may lay the foundation lead to light up this way.

Intraspecific differences in Androctunus crassicauda venom and envenomation symptoms.

Envenomation by Androctunus crassicauda is very frequent in Iran, especially in the south-west. This scorpion is one of the six scorpions whose venom is used to prepare anti-venom. Using HPLC, we discovered venom components of A. crassicauda varies from one specimen to another depending on geographical location, and this result is confirmed by those first found in various symptoms of A. crassicauda sting in envenomed persons from two separate geographical places (north and south of Khuzestan province). There was a significant relationship between symptoms and location of envenomation by A. crassicauda. Muscle spasm was more dominant in envenomed people from Northern cities, and venom chromatogram analysis showed the presence of at least six main sharp peaks in Northern A. crassicauda rather than Southern A. crassicauda. It shows intraspecific differences in venom of A. crassicauda that must be considered in treatment of stung people from different geographical locations as well as in the preparation of anti-venom. See also Figure 1(Fig. 1).

Presence probability of Hemiscorpius lepturus Peters, 1861 using maximum entropy approach in the western areas of Zagros Mountains, Iran.

AIM: The purpose of this research was to use environmental variables for predicting the probability of Hemiscorpius lepturus existence in the provinces where situated in the west of the Zagros Mountains. MATERIALS AND METHODS: In this study, 64 occurrence records of the H. lepturus were extracted from the published documents available in electronic databases. MaxEnt model was used for predicting the ecological niches of this species. Normalized difference vegetation index (NDVI) and 19 climatic variables were used as the environmental variables affecting the distribution of this scorpion. The Jackknife test in the model was used to indicate the importance of variables to predict the probability of the presence of the studied species. The logistic threshold that was evaluated using a logistic regression algorithm showed the converting of the probability model into a binary model. The model was evaluated byarea under the curve (AUC). The probability presence map of this scorpion was then prepared in ArcGIS 10.5 Software. RESULTS: The results of the analysis showed that the most important environmental factor on the distribution of H. lepturus was the maximum temperature of the warmest month (Bio5) with a contribution rate of 43% and permutation importance of 8%. The Jackknife test revealed that NDVI did not gain any value when it used independently in the model. The logistic threshold was reported 0.255 for the maximum test sensitivity plus specificity. The AUC of the model was 0.7698, shows an acceptable value for model validity. Overall the hot spots for this toxic scorpion seem to be in Khuzestan, Lorestan, and Ilam Provinces of the studied area. CONCLUSION: Regarding our findings, MaxEnt algorithm, in combination with geographic information system contributed to revealing the effects of environmental variables on the probability of H. lepturus presence in the west of Zagros Mountains. These visualized maps as a warning alarm can be helpful to policymakers for managing, controlling, and monitoring the scorpionism in high-risk areas.

A Comparative Study of Nesting Sites and Burrowing Habits of Two Iranian Burrowing Scorpions.

BACKGROUND: The different features of scorpions can be successfully described by their nesting and burrowing behaviors. There is little information about burrowing activity of Iranian scorpions. METHODS: The current study was performed to compare the burrowing behavior between two burrowing Iranian scorpions, Scorpio maurus and Odonthubutus bidentatus by describing 30 nests of each species regarding collecting the scorpions. RESULTS: Scorpio maurus and O. bidentatus have a tendency to make nest with elliptical, round-like entrance and oval shape with arch at the top, respectively. There was not any significant difference between nest entrance properties of two scorpions. One-way ANOVA test showed that the height and diameter of two species nests were not significantly different. A Pearson correlation also showed a relative strong direct relationship between height and diameter of S. maurus nests than O. bidentatus. This correlation was not significant in the case of O. bidentatus. The results provided additional habitat information of scorpions. CONCLUSION: The nests morphology characteristics of two Iranian scorpions including shape, depth, length and diameter depend are different from each other based on the following factors: species, soil texture, soil moisture and region conditions.

Molecular characterization of Leishmania parasites isolated from sandflies species of a zoonotic cutaneous leishmaniasis in Musiyan south west Iran.

Cutaneous leishmaniasis (CL) is vector borne parasitic disease, considered as public health problem especially in border of Iran and Iraq, Dehloran County (Musian district). The aim of this study was molecular identification of Leishmania parasites in sandfly as vectors of Leishmaniasis. Totally 280 female sandflies were trapped by sticky traps from 7 rural areas of Musiyan in September-November 2012. All sandflies were identified using morphological characters of the head and abdominal terminalia. DNA was extracted from female sandflies and Leishmania was identified using PCR and sequencing. All 280 trapped sandflies were identified as Phelobotumus Papatasi and Leishmania infections were detected in 3.2 % out of 280 female sandflies. All leishmania were identified as L. major and submitted in Gene bank as: LC014642.1, LC014641.1, LC014640.1 and LC014639.1. Frequency of Phlebotomus Papatasi and infection with L. major in studied regions showed that this vector is dominant in these areas.

Biocontrol of the Brown-Banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae), with Entomopathogenic Fungus, Metharhizium anisopliae.

BACKGROUND: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formulations of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longipalpa F. under laboratory and field conditions. METHODS: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 10(8) conidia ml(-1) followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs using different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer. RESULTS: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough pathogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in-water formulation was more effective than the bait formulation against the cockroach caused 39.4-97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sunflower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses. CONCLUSION: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising alternative for cockroach control.

Assessment of immunogenic characteristics of Hemiscorpius lepturus venom and its cross-reactivity with venoms from Androctonus crassicauda and Mesobuthus eupeus.

Hemiscorpius lepturus (H. lepturus), one of the most venomous scorpions in tropical and sub-tropical areas, belongs to the Hemiscorpiidae family. Studies of antibodies in sera against the protein component of the venom from this organism can be of great use for the development of engineered variants of proteins for eventual use in the diagnosis/treatment of, and prevention of reactions to, stings. In the present in vitro study, the proteins of H. lepturus venom, which could specifically activate the production of immunoglobulin G (IgG) in victims accidently exposed to the venom from this scorpion, were evaluated and their cross-reactivity with venoms from two other important scorpion species including Androctonus crassicauda and Mesobuthus eupeus assessed. H. lepturus venom was analyzed with respect to its protein composition and its antigenic properties against antibodies found in sera collected from victims exposed to the venom of this scorpion within a previous 2-month period. The cross-reactivity of the H. lepturus venom with those from A. crassicauda and M. eupeus was assessed using ELISA and immunoblotting. Electrophoretic analysis of the venom of H. lepturus revealed several protein bands with weights of 8-116 KDa. The most frequent IgG-reactive bands in the test sera had weights of 34, 50, and 116 kDa. A weak cross-reactivity H. lepturus of venom with venoms from A. crassicauda and M. eupeus was detected. The results of immunoblotting and ELISA experiments revealed that H. lepturus venom activated the host immune response, leading to the production of a high titer of antibodies. Clearly, a determination of the major immunogenic components of H. lepturus venom could be valuable for future studies and ultimately of great importance for the potential production of recombinant or hypo-venom variants of these proteins.

A climatological study of scorpion sting incidence from 2007 to 2011 in the Dezful area of southwestern Iran, using a time series model.

Scorpion stings are a public health problem in south and southwest Iran. There is little information regarding climatological effects on incidence of scorpion stings in Iran. Therefore, the present systemic survey of scorpion sting data was conducted from the point of view of entomo-meteorological relationships and analyzed statistically for the Dezful area in Khuzestan, southwest of Iran. The time series analysis was implemented using MINITAB version 16 statistical software packages. In total, 3,755 scorpion sting files from the Dezful health centers were monitored from April 2007 to September 2011 in a time series analysis. The results showed that temperature had significant effects on scorpion sting. From the data of this study, it is concluded that the scorpion activity in Dezful County is a climatological-dependent phenomenon.

Evaluation of Conidia-Dust Formulation of the Entomopathogenic Fungus, Metarhizium anisopliae to Biocontrol the Brown-Banded Cockroach, Supella longipalpa F.

BACKGROUND: The brown-banded cockroach Supella longipalpa (F.) as a mechanical vector of pathogens and source of allergens has recently become widespread in the city of Ahvaz, southwestern Iran. OBJECTIVES: This research was done to evaluate the efficacy of a dust-formulation of Metarhizium anisopliae isolate IRAN 437C, as a common entomopathogenous fungus, against S. longipalpa. MATERIALS AND METHODS: Conidia dust-formulations of M. anisopliae were prepared in proportions of 1%, 5%, 10%, 25%, 50% and 100% with bad wheat flour as the carrier. Cockroaches were exposed to surfaces treated with 1.5 mg/cm(2) of the formulations under laboratory and semi-field conditions. RESULTS: Cockroach mortality rates increased and survival times (ST50) decreased with an increased proportion of conidia from 1% to 100% but records taken for mortality and survival time from proportions of 25%, 50% and 100% were not significantly different. The mortality rates reached 100% and 90-100% in adults and nymphs, respectively on the seventh day. The lowest ST50 was related to the proportion of 100% (3 days). Probit analysis indicated LD50 and LD90 values of 1.7 × 10(6) and 1.7 × 10(7) conidia/cm(2) for adults and these values changed to 4.5 × 10(6) and 2.9 × 10(7) for third and fourth instar nymphs at three days post exposure. Proportion of 25% caused mortality rates of 87%, 81% and 73% in adult, adult & nymph and nymph populations, respectively at four days after exposure under room conditions. CONCLUSIONS: Conidia dust-formulation of M. anospliae isolate IRAN 437C could present a promising alternative to control the brown-banded cockroach.

Culiseta subochrea as a bioindicator of metal contamination in Shadegan International Wetland, Iran (Diptera: Culicidae).

The quantity of some trace metals of mosquito larvae in Shadegan International Wetland from Iran was evaluated. Water, waterbed sediment, and mosquito larvae samplings were carried out from an urban site in the east of the wetland, using standard methods in December 2011. The identified Culiseta subochrea (Edwards) and Aedes caspius s.l. (Pallas) larvae, water, and waterbed sediment samples were analyzed for As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Pb, and Zn trace metals using standard preparation and isolation procedure. Result showed that the waterbed sediment and Cu. subochrea larvae are polluted with all trace metals investigated except As and Hg. The trace metals bioaccumulated in the Cu. subochrea larvae range from 31.78 at the lowest level for Cr to 3822.7 at the highest level for Cd. In a conclusion, this is the first report confirmed that Cu. subochrea likely used as a bioindicator to trace metal pollution in marine ecosystems in the world, especially wetlands.

The First Report of Drug Resistant Bacteria Isolated from the Brown-Banded Cockroach, Supella longipalpa, in Ahvaz, South-western Iran.

BACKGROUND: The brown-banded cockroach, Supella longipalpa is known as a carrier of pathogenic bacteria in urban environments, but its role is not well documented regarding the carriage of antibiotic-resistant pathogenic bacteria in Iran. The aim of this study was to determine the resistance bacteria isolated from the brown-banded cockroach in Ahvaz, south west of Iran. METHODS: Totally 39 cockroaches were collected from kitchen area of houses and identified. All specimens were cultured to isolate the bacterial agents on blood agar and MacConky agar media. The microorganisms were identified using necessary differential and biochemical tests. Antimicrobial susceptibility tests were performed for isolated organisms by Kirby-Bauer's disk diffusion according to NCLI guideline, using 18 antibiotics. RESULTS: From the 39 collected S. langipalpa, 179 bacterial agents were isolated, 92 of alimentary ducts and 87 of external body surfaces. Isolated bacteria from cockroaches were identified as Enterobacter spp., Klebsiella spp., Citrobacter spp., Escherichia coli, Salmonella spp., Proteus spp., coagulase negative staphylococci, Serratia marcescens, Staphylococcus aureus, and Bacillus species. The pattern resistance rates were determined for gram negative bacilli and gram positive cocci regarding 18 antibiotics. CONCLUSION: The brown-banded cockroach can be involved in the spread of drug resistant bacteria and increases the possibility of contacting human environment to drug resistant bacteria. Therefore, the potential of removing this insect should be improved. This is the first original report of drug resistant bacteria isolated from the brown-banded cockroach of Iran.

The roles of some scorpions, Hemiscorpius lepturus and Androctonus crassicauda, in a scorpionism focus in Ramhormorz, southwestern Iran.

Scorpion stings are a common and important health problem in Iran, particularly in south and southwestern Iran, including the province of Khuzestan. In the area of Khuzestan near the city of Ramhormoz, Hemiscorpius lepturus (Scorpionida: Hemiscorpioiidae) and Androctonus crassicauda (Buthidae) are present. Ramhormoz is in southwestern Iran and is one of the most important foci of the scorpion sting problem. The current study was carried out to gain both epidemiological and medical information about scorpion stings in and around the city of Ramhormoz. In total, 179 people who were admitted to the Emergency Department of Ramhormoz Imam Khomeini Hospital during 2008 and 2009 after being stung by scorpions were monitored. Epidemiological and medical parameters including sex of the victim; the part of the body stung; the month when stung; the biochemical parameters comprising blood sugar (BS), blood urea nitrogen (BUN), and creatinine (CR); hematological parameters including white blood cells (WBC), count blood cells (CBC), red blood cells (RBC), hemoglobin (Hb), hematocrit (HCT), platelet (PLT); and urine analysis including hemoglobinuria were recorded. The current study showed that most of the victims were stung by H. lepturus, while very few were stung by A. crassicaud, but in over half of the cases the species was not known. Stings were most common from May to Aguust. 73% of the victims were female. The limbs were the part of the body most likely to be stung. Hemogobinuria was very common in H. lepturus victims.

Study on the heavy metal bioconcentrations of the Shadegan international wetland mosquitofish, Gambusia affinis, by inductively coupled plasma technique.

The purpose of this study was to evaluate the levels of heavy metal bioconcentration of the mosquitofish (Gambusia affinis) in Shadegan international wetland. Sampling including the water, waterbed sediment and mosquitofish was carried out from the selected sampling sites during October and November 2011, and analyzed by the ICP-OES. Results show that the water has poor qualitative condition, according to EPA and WHO water quality standards. The level of the water Cr in the selected sites in both months and the levels of Fe, Mn and Zn during October in the SW1 site were higher than the instrumental detection limits indicating that the water was contaminated with these metals in the mentioned sites and months. The levels of the waterbed sediment As, Co, Cr, Cu, Fe, Mn, Pb and Zn, and mosquitofish Cr, Cu, Fe, Mn, Zn, Co and Cd were much higher than the instrumental detection limits, indicating that the waterbed sediment and mosquitofish were contaminated with them during October and November in the selected sites. Statistical assessments reveal that there is a significant difference between the mentioned contaminated water, waterbed sediment and mosquitofish heavy metals (all P-values < 0.05). In overall, it is considered that the contaminated heavy metals can be accumulated in the waterbed sediment and bioconcentrated in the wildlife tissues, then finally can be entered in the marine food chains and biomagnified there after long periods. In conclusion, this paper confirmed that the G. affinis can be used as a bioindicator of heavy metal pollution in marine ecosystems such as wetlands.

An in vitro comparative study upon the toxic properties of the venoms from Hemiscorpius lepturus, Androctonus crassicauda and Mesobuthus eupeus scorpions.

The aim of the present study was to compare the toxic effects of the venoms from Hemiscorpius lepturus (H. lepturus), Androctonus crassicauda (A. crassicauda) and Mesobuthus eupeus (M. eupeus). For this purpose, three in vitro models were employed to compare the toxic effects of various concentrations of the venoms from these three scorpions, namely: hemolytic potential using human RBCs, phospholipase activity using Saubouraud's dextrose agar (SDA) supplemented with 2% egg yolk and lactate dehydrogenase (LDH) enzyme releasing effect using K562 leukemia cell line. In addition, the neutralizing effectiveness of the antivenom against these toxic properties was assessed. The results showed that, unlike the venoms from A. crassicauda and M. eupeus, the venom from H. lepturus produced dose-dependent lysis of human RBCs and showed phospholipase activity. However, all the tested venoms showed variable degrees of LDH releasing properties. The venom from H. lepturus had highest and the venom from M. eupeus had the lowest LDH releasing effect. The antivenom effectively inhibited all the tested toxicities. In conclusion, these results suggest that the venoms from the studied scorpions have variable toxic properties, which may explain the underlying reason for the differences in their clinical manifestations. In addition, the antivenom was effective in neutralizing all the tested toxic effects.

Orbital myiasis.

PURPOSE: To present a case of massive orbital myiasis. CASE REPORT: An 87-year-old debilitated woman suffering from left ocular pain of four days' duration presented with a severely necrotized left orbit and several attached live larvae. The upper and lower eyelids and the eyeball were completely destroyed. She had history of eyelid surgery in the same eye due to a skin lesion, apparently some type of skin cancer, 15 years before. The larvae were identified as Chrysomya bezziana (Diptera: Calliphoridae) or old world screwworm fly. CONCLUSION: Infestation of ocular and orbital tissues by fly larvae (ophthalmomyiasis) progresses rapidly and can completely destroy orbital tissues within days, especially in patients with poor general health. Treatment consists of removal of the larvae and surgical debridement.