An Experimental Study of the Effects of Mesobuthus eupeus Scorpion Venom on Plasma Concentrations of Metabolic Hormones and Glucose in Rats

Document Type: Clinical Pathology

Authors

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz,Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Department of Venomous Animals and Toxins, Razi Vaccine and Serum Research Institute, Ahvaz, Iran

Abstract

BACKGROUND: Mesobuthus eupeus is an indigenous scorpion species in Southwest Iran which is respon- sible for the majority of scorpion sting cases in Khuzestan province.
OBJECTIVES: The effects of M. eupeus venom were investigated on the chief metabolic hormones in rats.
METHODS: A total of 45 Albino male rats were divided into 3 equal groups: group 1 (control): 0.5 ml normal saline was administered intraperitoneally (IP); group 2 and 3: M. eupeus venom was administered with a dose of 1 and 2 mg/kg IP, respectively. Sampling was performed at 8, 24, and 48 hours after venom/ saline injection.
RESULTS: The levels of thyroxine (T4) and triiodothyronine (T3) were significantly lower in both venom receiving groups (groups 2 and 3) than in the control group, dose-dependently, at all sampling times. There was a significant decrease in insulin level in both intoxicated groups compared to the control group at all sampling times. Glucagon, cortisol and subsequently glucose concentrations were significantly increased in both groups receiving the venom (groups 2 and 3) compared to the control group at 8 and 24 hours following envenomation.
CONCLUSIONS: The findings of this study indicate that M. eupeus venom can suppress the secretion of essential metabolic hormones including T3, T4, and insulin and stimulate the release of glucagon, and cor- tisol, leading to hyperglycemia.

Keywords


Article Title [Persian]

مطالعه تجربی اثرات سم عقرب مزوبوتوس اوپئوس بر غلظت پلاسمایی هورمون‌های متابولیک و گلوکز در موش صحرایی

Authors [Persian]

  • محمد راضی جلالی 1
  • سیده میثاق جلالی 2
  • هدیه جعفری 3
  • محمد باباخان 2
1 گروه علوم درمانگاهی، دانشکده دامپزشکی دانشگاه شهید چمران اهواز، اهواز، ایران
2 گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه شهید چمران اهواز، اهواز، ایران
3 موسسه تحقیقات واکسن و سرم سازی رازی، اهواز، ایران
Abstract [Persian]

 
 
زمینه مطالعه: مزوبوتوس اوپئوس یک گونه عقرب بومی در جنوب غربی ایران است که مسئول اکثر موارد عقرب گزیدگی در استان خوزستان می‌باشد.

هدف: اثر سم عقرب مزوبوتوس اوپئوس بر هورمون‌های متابولیک اصلی در موش صحرایی مورد بررسی قرار گرفت.

روش کار: 45 سر موش صحرایی نر آلبینو به 3 گروه مساوی تقسیم شدند: گروه 1 (کنترل): 5/0 میلی‌لیتر نرمال سالین به صورت داخل‌صفاقی دریافت کردند، گروه 2 و 3: سم عقرب مزوبوتوس اوپئوس به ترتیب با دوز 1 و 2 میلی‌گرم به ازای هر کیلوگرم به صورت داخل صفاقی دریافت نمودند. نمونه‌گیری در زمان 8، 24 و 48 ساعت پس از تزریق سم و یا سالین انجام گرفت.

نتایج: مقادیر تیروکسین (T4) و تری‌یدوتیرونین (T3) در هر دو گروه دریافت کننده سم (گروه 2 و 3) نسبت به گروه کنترل در همه زمان‌ها به صورت وابسته به دوز به طور معنی‌داری پایین‌تر بود. همچنین کاهش معنی‌داری در میزان انسولین در هر دو گروه دریافت کننده سم در مقایسه با گروه کنترل در همه زمان‌ها مشاهده گردید. غلظت گلوکاگن، کورتیزول و گلوکز در هر دو گروه دریافت کننده سم (گروه 2 و 3) در مقایسه با گروه کنترل پس از 8 و 24 ساعت به‌طور معنی‌داری افزایش یافته بود.

نتیجه‌گیری نهایی: یافته‌های این مطالعه نشان داد سم عقرب مزوبوتوس اوپئوس قادر به سرکوب ترشح هورمون‌های متابولیک اصلی شامل T3 و T4 و انسولین و تحریک ترشح گلوکاگن و کورتیزول می‌باشد که در مجموع منجر به هیپرگلایسمی می‌شوند.

Keywords [Persian]

  • مزوبوتوس اوپئوس
  • عقرب
  • سم
  • سیستم درون‌ریز
  • انسولین
  • گلوکاگن

Introduction

Scorpion sting is one of the major health challenges for humans and animals, espe- cially in tropical and developing countries. More than 1,500 scorpion species have been identified worldwide (Lourenço, 2014; Lu- cas and Meier, 2017) and about 32 species were found in Iran, 12 of which have been detected in Khuzestan (Jalali and Rahim 2014; Rahimi et al., 2015).

The Buthidae are the largest family of scorpions and are found all over the world especially in tropical and semi-tropical re- gions, particularly the Middle East and Cen- tral Asia (Lourenço, 2014; Lucas and Meier, 2017). Mesobuthus eupeus is the most abun- dant species in Iran which is found in Khuz- estan province and is responsible for 21.7% of cases of scorpion sting in Khuzestan (Ra- himi et al., 2015; Dehghani et al.,  2017).

The venom of scorpions, especially the Buthidae family, consists of a variety of toxins and active biological compounds (Khoobdel et al., 2013; Erdeş et al., 2014; Laraba-Djebari et al., 2015) which can af- fect various tissues (Murthy and Zare, 2001; Murthy and Zare, 2002; Razi Jalali et al., 2015). Scorpion envenomation can lead  to the failure of many organs and major met- abolic and endocrine disorders through the release of large amounts of catecholamines, and vascular disorders (Petricevich,  2010). In some studies, scorpion stings were found to be associated with alterations in insulin, glucagon, and glucocorticoid secretion level, and in blood glucose levels (Murthy, 2000). However, there are few studies about the ef- fect of scorpion venom on thyroid secretion (Murthy and Zare, 1998).

To the best of the authors' knowledge,  there has been no documented data on the effects  of  M.  eupeus  venom  on  the endo-


 

crine system, including thyroid, endocrine pancreas and adrenal secretions. Since it is a native Iranian species, research into various aspects of the pathogenesis of its venom are important to improve diagnostic and thera- peutic methods. Therefore, the present study was designed to investigate the effects of Mesobutupus eupeus envenomation on plas- ma thyroid hormones (T3 and T4), insulin, glucagon, cortisol, and glucose levels in rats.

Materials and Methods

Scorpion venom

The venom of M. eupeus scorpion venom used in this study was provided by Razi Vac- cine and Serum Research Institute, Ahvaz which was prepared by electric shock and kept in lyophilized form. The concentration of crude venom protein was 815 mg/gven- om. Solutions of 0.05% and 0.1% of venom were prepared in 0.9% sodium chloride solu- tion in suitable volumes proportional to the number of rats in each envenomated group, as described below.

Laboratory animals

A  total  of  45  Albino  male  rats  (Wis- tar strain)  weighing  250–300  g aged  4 to   6 months were housed in groups of six, in plastic cages, in an air-conditioned room maintained at a temperature of 24 ± 2 °C and a relative humidity of 55 ±5%, with a 12-h light/12-h dark illumination cycle.  They were fed a commercial laboratory pellet diet and tap water ad libitum. All procedures were done in accordance with ethical guide- lines for care and use of laboratory animals, discarding of dead  animals  and  protection of the researcher against animal bites and were approved by the Experimental Animals Committee of Shahid  Chamran  University of Ahvaz, Iran.

 

 

 

Experimental design

Animals were randomly divided into 3 equal groups and were treated as  follows:

Group 1: (control group) 0.5 ml normal sa- line intraperitoneally (IP).

Group 2: M. eupeus  venom  at a dose  of  1 mg/kg body weight IP (0.5 ml of 0.05% solution for each rat).

Group 3: M. eupeus venom at a dose of 2 mg/kg body weight IP (0.5 ml of 0.1% solu- tion for each rat).

The doses of venom were selected based on previous studies by authors (Razi Jalali et al., 2015; 2016).

Blood collection

Sampling was performed at 8, 24, and 48  h after venom/saline injection; five rats from each group were sampled each time. Blood samples were collected with heparin via cardiac puncture following anesthesia with chloroform (Merck, Germany). After centrif- ugation the plasma was separated and stored at -20 ºC for subsequent measurements.

Plasma biochemical  analysis

The plasma levels of T4 and T3 were mea- sured using ELISA kits (Sigma-Aldrich). Cortisol, insulin and glucagon were also assessed  by  ELISA  (Monobind,  Germany)


according to the manufacturer's instructions. The amount of glucose was measured using colorimetric kits (Pars Azmun, Iran) via bio- chemical autoanalyzer (Biotecnica, Bt-1500, Italy).

Statistical analysis

Analysis of variance and Tukey post-hoc tests were employed to compare the data be- tween groups using SPSS software version  16 (SPSS Inc., Chicago, Illinois, USA). All values were expressed as mean ± standard error, and p < 0.05 was considered as statisti- cally significant.

Results

Clinical signs

The envenomated rats showed behavioral changes from aggression to depression. In addition, hemorrhage and hyperemia in the eyes and mucosal membranes were recorded in both intoxicated groups.

T3 measurement

The results of T3 evaluation showed that the levels of this hormone were significantly lower in group 2 than in the control group and in group 3 compared to the rest of the groups, at all sampling times (p <0.05) (Ta- ble 1). In addition, in group 3, the amount of

 

 

Table 1. Plasma triiodothyronine (T3) and thyroxine (T4) concentra- tion as mean ± SE in different groups and sampling times.

 

Group

T3 (µg/dl)

T4 (µg/dl)

h 8

h 24

h 48

h 8

h 24

h 48

 

Group 1   (control)

0.08 ± 2.48

 

*A

0.11 ± 2.35

 

A

0.09 ± 2.32

 

A

1.33 ± 23.33

 

*A

1.31 ± 24.75

 

A

1.93 ± 24.25

 

A

 

Group 2   (venom 1 mg/kg)

0.05 ± 1.34

 

B

0.05 ± 1.46

 

B

0.09 ± 1.91

 

B

1.53 ± 18.20

 

B

1.28 ± 21.40

 

B

1.39 ± 22.80

 

A

 

Group 3   (venom 2 mg/kg)

0.04 ± 0.97

 

**Ca

0.13 ± 1.05

 

Cb

0.12 ± 1.98

 

Bc

1.48 ± 17.02

 

B

1.35 ± 20.20

 

B

1.69 ± 22.40

 

A

* Different uppercase letters in each column represent significant difference between groups.

** Different lowercase letters in each row represent significant difference between sampling times.

 

 

 

 

 

 

 

Group


Table 1. Plasma triiodothyronine (T3) and thyroxine (T4) concentra- tion as mean ± SE in different groups and sampling times.

Insulin (µIU/dl)                                                      Glucagon (ng/ml)

 

                       

8 h                     24 h                    48 h                     8 h                     24 h                    48 h

 

 

Group 1 (control)

 

Group 2 (venom 1 mg/kg)

 

Group 3 (venom 2 mg/kg)


12.70 ± 0.92

A* 8.40 ± 1.08

B 8.13 ± 0.74

B


12.13 ± 1.81

A 9.36 ± 1.29

B 7.06 ± 1.29

B


11.40 ± 1.92

A 10.60 ± 1.20

B 8.84 ± 1.11

B


123.67 ± 2.60

A* 149.20 ± 2.70

Ba** 156.50 ± 2.46

Ca


122.25 ± 2.27

A 154.40 ± 0.67

Bb 168.80 ± 1.02

Cb


121.50 ± 1.19

A 139.80 ± 1.70

Ac 136.00 ± 1.81

Ac

 

 

 

* Different uppercase letters in each column represent significant difference between groups.

** Different lowercase letters in each row represent significant difference between sampling times.

 

 

T3 significantly increased over time, so that the highest hormone level was at 48 h post injection (p <0.05).

T4 measurement

Based on the results of T4 measurement (Table 1), the hormone levels decreased in both envenomated groups  (groups  2 and 3) in comparison with the control group, so that the difference was significant at 8 and 24 h (p <0.05). However, with the passage of time at 48 h after venom injection, this decrease was moderated and there was no significant difference between groups at any of the sam- pling times.

Insulin measurement

Based on the results (Table 2), there was a significant decrease in insulin level in the in- toxicated groups (groups 2 and 3) compared to the control group at all sampling times (p<0.05).

Glucagon measurement

The administration of venom signifi- cantly increased glucagon level in group 2 compared to the control group and in group  3 compared to the previous two groups at 8 and 24 h post injection (p <0.05) (Table 2). Additionally, in both intoxicated groups, the maximum amount of hormone was observed at 24 h post injection (p <0.05).


Cortisol measurement

A significant increase in cortisol concen- tration was noted in both groups receiving the venom (groups 2 and 3) compared to the control group at 8 and 24 h following en- venomation (p<0.05) (Table 3). However, in both mentioned groups, the level of cortisol decreased significantly at 48 h in compari- son to the previous sampling times (p <0.05).

Glucose measurement

Glucose levels in both venom injected groups (groups 2 and 3) were significantly increased compared to the control group at  all sampling times (p <0.05) (Table 3). How- ever, there was no significant difference be- tween two envenomed groups in terms of glucose levels.

Discussion

In the present study, injection of M. eupeus venom resulted  in  a  significant  reduction  in the thyroid hormones (T3 and T4). The highest decrease was observed in the third group (high dose venom) at 8 h after injec- tion, considering the half-life of T3 which is only 1 day while the half-life of T4 is 5-7 days. These changes might be attributable to an autonomic storm through the release of catecholamines and angiotensin II  following

 

 

Table 1. Plasma triiodothyronine (T3) and thyroxine (T4) concentra- tion as mean ± SE in different groups and sampling times.

 

 

 

 

 

 

 

 

 

 

 

           

   
                                                                                                                                                                                                                                             
     

 

     

Group

     
     

Cortisol       (µg/dl)

     
     

Glucose       (mg/dl)

     
     

8 h

     
     

24 h

     
     

48 h

     
     

8 h

     
     

24 h

     
     

48 h

     
     

 

     

Group 1       (control)

     
     

2.36 ±       0.11

     

A*

     
     

3.11 ±       0.39

     

A

     
     

2.41 ±       0.55

     

A

     
     

86.67 ±       6.61

     

A*

     
     

84.25 ±       4.20

     

A

     
     

87.50 ±       5.52

     

A

     
     

 

     

Group 2       (venom 1 mg/kg)

     
     

9.34 ±       1.17

     

Ba**

     
     

8.74 ±       0.75

     

Ba

     
     

4.20 ±       0.27

     

Ab

     
     

129.00 ±       4.78

     

B

     
     

138.60 ±       3.45

     

B

     
     

126.40 ±       4.41

     

B

     
     

 

     

Group 3       (venom 2 mg/kg)

     
     

9.82 ±       1.29

     

Ba

     
     

8.63 ±       0.75

     

Ba

     
     

3.92 ±       0.26

     

Ab

     
     

138.00 ±       3.08

     

B

     
     

145.60 ±       6.08

     

B

     
     

134.40 ±       3.70

     

B

     
   

 

   
   

* Different uppercase letters in each column represent significant difference between groups.

 

** Different lowercase letters in each row represent significant difference between sampling times.

 

 

envenomation, which led to a decrease in in- sulin secretion and subsequent reduction of T3 and T4 concentration in the blood. These findings are consistent with other previous studies in this regard. In a study performed by Murthy and Zare (1998) a significant decrease in T4 and T3 levels was observed following the injection of Mesobuthus ta- mulus venom in dogs. They also obtained similar results regarding  T3  concentration in rabbits. In addition, while anti-venom ad- ministration resulted in an increase in serum T3 level in dogs, it could not prevent venom induced hormonal changes in rabbits. How- ever, the intracerebroventricular injection of Kaliotoxin (KTX) fraction of Androctonus australis venom, in another  study,  resulted in the stimulation of neuro-endocrine re- sponse with significant rise in serum T3, T4, and TSH levels at 24 h post-injection in rats which was associated with inflammatory cell infiltration and imbalanced redox status in both hypothalamus and thyroid tissue (Lad- jel-Mendil et al., 2016). This diversity in the results might be due to the difference in the type of studied scorpion species and venom as well as the route of venom administration. According to the results of this study, glu- cagon levels were significantly increased   in


the venom treated groups at 8 and 24 h after injection in a dose-response manner. How- ever, the levels of this hormone returned to normal in both groups at 48 h after enven- omation. This effect is probably mediated through the adrenergic stimulation of endo- crine pancreas as well as other tissues. Sim- ilar alterations in glucagon secretion were also reported by Johnson and Ensinck (1976) following pancreas perfusion with Leiurus quinquestriatus scorpion toxin in rats. This reduction in glucagon secretion was associat- ed with the stimulation of sympathetic nerve terminals in pancreas tissue and increased norepinephrine release which was success- fully prevented in the presence of alpha and beta-adrenergic blocking agents in rat. In ad- dition, subcutaneous administration of Me- sobuthus tamulus scorpion venom resulted in an increased blood glucagon in dogs. These hormonal changes seem to result in an im- balance in the carbohydrate metabolism and the inability of vital organs to use metabolic substrates (Murthy and Haghnazari,  1999).

The present study revealed that there was  a significant decrease in insulin levels in the envenomated rats compared to the control group, with the highest reduction at 8 and 24 h after  venominjection.  In other experimen-

 

 

 

tal studies, hypoinsulinemia or insulin resis- tance was reported following  the  injection of various scorpion spp. venom. During the study conducted by Murthy and Haghnazari (1999) and Zare et al. (1994), the same al- terations in insulin level was recorded fol- lowing Mesobuthus tamulus venom admin- istration in dogs. Furthermore, Androctonus australis hector scorpion toxin was associat- ed with severe inflammatory response, liver tissue damage and hyperglycemia accompa- nied by hyperinsulinemia, indicative of insu- lin resistance in a previous study in rats. It is also possible that the insulin measured after venom injection was not functionally active. Moreover, pretreatment with cytokine antag- onists, in the same study, significantly de- creased inflammatory biomarkers and plas- ma glucose levels as well as hepatic tissue damage and metabolic disorders which was suggestive of the crucial role of inflammato- ry cytokines (IL-6 and TNF-α) in the com- plications induced by the scorpion venom (Taibi- Djennah and Laraba-Djebari,  2015).

In addition, exogenous insulin admin- istration was demonstrated to reverse he- modynamic, cardiovascular, metabolic, electro-cardiographic (ECG) changes and pulmonary oedema in the experimental ani- mals and in the scorpion sting victims which represents the key role of insulin suppres- sion in the pathogenesis of the toxin and also suggests that the insulin receptor and the signaling pathways are not defective (Mur- thy, 2013, 2014a; Murthy et al., 1988, 1990, 1991; Yugandhar et al., 1999). It appears that scorpion envenomation, including M. eu- peus, results in a substantial release of cat- echolamines and Angiotensin II which can inhibit glucose-induced secretion of insulin from the beta cells of islets in the endocrine pancreas as well as antagonizing the  actions


of insulin by promoting glycogenolysis which leads to hyperglycemia. The suppres- sion of insulin secretion or insulin resistance along with a rise in catabolic counter-regu- latory hormones including glucagon, corti- sol and glucose level might be responsible for the pathogenesis of a variety of clinical symptoms. Consequently, M. eupeus venom, like many other scorpion species, can induce a syndrome of energy  scarcity  and inabili-  ty of vital organs to consume the metabolic substrates causing multi-organ system fail- ure (Murthy, 2014b).

In the current experiment, the injection of scorpion venom in both groups led to a sig- nificant increase in cortisol levels. The high- est hormone concentration was observed at 8 h after receiving the toxin and decreased over time, so that cortisol changes were not sig- nificant within 48 h. Other previous studies have found similar findings. Subcutaneous administration of M. tamulus resulted in an increased blood cortisol in dogs and rabbits (Murthy and Haghnazari, 1999; Zare et al., 1994). Excessive release of catecholamines following scorpion venom intoxication is, once again, likely responsible for increased endogenous glucocorticoids which,  in turn, is often associated with carbohydrate intol- erance (Murthy, 2014b).

The results of glucose measurements  in the present study showed a significant in- crease in the level of this analyte after in- jection of M. eupeus venom at all sampling times compared to control group. Consider- ing that there was no significant difference between the two groups of venom recipi- ents, it seems that changes in glucose do not correlate with the dose of injected venom. Hyperglycemia has already been observed following injection of various scorpion spp. toxins including Mesobuthus tumulus   (Mur-

 

 

 

thy and Haghnazari; 1999 Zare et al., 1994), Androctonus australis hector (Taibi-Djennah and Laraba-Djebari, 2015), Tityus serrula- tus (Andrade et al., 2004), and Indian black scorpion (More et al., 2004). These effects are most likely due to the increased circu- lating levels of catabolic counter-regulatory hormones, including glucagon, cortisol and epinephrine, which act synergistically  as well as simultaneous suppression in insulin secretion that induce sustained hepatic glu- cose production and inability of tissues to utilize it.

In this study, the venom injected rats showed some behavioral changes from ag- gression to depression.  This  might  be  due to the effects of venom on central nervous system. Although this issue is not directly relevant to the present study, there is consid- erable evidence showing the direct partici- pation of the central nervous system in the envenoming process provoked by scorpions (Sadeghian, 2003; Nencioni et al., 2018)  with numerous proposed mechanisms in- cluding: alterations in coagulation of blood leading to disseminated intravascular coagu- lation; high level of catecholamines induced, vasospasm causing hypoperfusion, and isch- emia in previously compromised areas of the brain; high blood pressure during autonomic storm resulting in rupture of vessels, caus- ing hemorrhagic stroke; and the presence of myocarditis, thromboembolic phenomenon, or shock leading to cerebral infarction (Sen- gupta et al., 2009; Thomas et al.,  2017).

In addition, hemorrhage  and  hyperemia  in the eyes and mucosal membranes were recorded in both intoxicated groups which might be attributed to a possible coagulopa- thy following scorpion intoxication (Rahma- ni and Jalali, 2012; Seyedian et al., 2014). However, in this study, laboratory tests  were

 

not carried out to confirm this.

In conclusion, the findings of this study indicate that M. eupeus venom  can  affect the endocrine system by suppressing the secretion of essential metabolic hormones including T3, T4, and insulin and excessive release of glucagon, and cortisol, leading to hyperglycemia and disorders in the overall metabolism of the body. These alterations might be responsible for the pathogenesis of a variety of clinical symptoms following en- venomation. However, the identification of other aspects and mechanisms involved  in the toxicity of this scorpion species venom requires more research.

Acknowledgments

The authors would like to thank Deputy of Research of Shahid Chamran University of Ahvaz for funding this study.

Conflict of Interest

The authors declare that there is no con- flict of interest.

Andrade, M.V., Caramez, M.P.R., Abreu, E.M.N.,

Dolnikoff, M., Omar, E.D., Velasco, I.T., Cunha-Melo, J.R. (2004) Lung compliance, plasma electrolyte levels and acid–base bal- ance are affected by scorpion envenomation in anesthetized rats under mechanical ventilation. Comp Biochem Physiol C Pharmacol  Toxi- col Endocrinol, 138(1), 97-104. https://doi. org/10.1016/j.cca.2004.05.009

Dehghani, R., Rafinejad, J., Fathi, B., Shahi, M.P., Jazayeri, M. and Hashemi, A. (2017). A retro- spective study on Scropionism in Iran (2002– 2011). J Arthropod Borne Dis, 11(2), 194-203.

PMID: 29062844

Erdeş, E., Doğan, T.S., Coşar, İ., Danışman, T., Kunt,  K.B.,  Şeker,  T.,  Yücel,  M.  and Özen,

C. (2014). Characterization of Leiurus abdul- lahbayrami   (Scorpiones:   Buthidae)  venom:

 

 

 

peptide profile, cytotoxicity and antimicrobial activity. J Venom Anim Toxins Incl Trop Dis, 20(1),     48.   https://doi.org/10.1186/1678-

Jalali, A. and Rahim, F. (2014). Epidemiological review of scorpion envenomation in Iran. Iran J Pharm Res, 13(3), 743–756. PMID: 25276176

Johnson, D.G., Ensinck, J.W. (1976). Stimulation of glucagon secretion by scorpion toxin in the perfused  rat  pancreas.  Diabetes,  25(8),  645-

Khoobdel, M., Zahraei-Salehi, T., Nayeri-Fasaei, B., Khosravi, M., Omidian, Z., Motedayen, M.H., Akbari, A. (2013). Purification of the Immunogenic  Fractions  and  Determination of Toxicity in Mesobuthus eupeus (Scorpioni- da: Buthidae) Venom. J Arthropod Borne Dis, 7(2),139-146.   PMID: 24409439

Ladjel‐Mendil, A., Martin‐Eauclaire, M.F., Lar- aba‐Djebari, F. (2016). Neuro‐Modulation of Immuno‐Endocrine Response Induced by Kali- otoxin of Androctonus Scorpion Venom. J Bio- chem Mol Toxicol, 30(12), 580-587. https:// doi.org/10.1002/jbt.21824

Laraba-Djebari, F., Adi-Bessalem, S. and Ham- moudi-Triki, D. (2015). Scorpion venoms: pathogenesis and biotherapies. In: Gopal- akrishnakone, P., Possani, L.D., Schwartz, E.F. and Rodríguez de la Vega, R.C. (eds), Scorpi- on  Venoms,  Toxinology  book  series, volume

4. Springer, Dordrecht. pp 63-85. https://doi. org/10.1007/978-94-007-6404-0_2

Lourenço, W.R.  (2014).  A  historical  approach to scorpion studies with special reference to  the 20 th and 21st centuries. J Venom Anim Toxins Incl Trop Dis, 20(1), 8. https://doi. org/10.1186/1678-9199-20-8

Lucas, S.M. and Meier, J. (2017). Biology and dis- tribution of scorpions of medical importance. In: Handbook of Clinical Toxicology of Animal Venoms and Poisons. White, J, and Meier, J. (eds), 1st ed. CRC Press. pp. 205-219. https:// doi.org/10.1201/9780203719442

More,  S.S.,  Kiran,  K.M.,  Gadag,  J.R.    (2004).

Dose-dependent serum biochemical alter- ations  in  Wistar  albino  rats  after Palamneus


gravimanus (Indian black scorpion) envenom- ation. J Basic Clin Physiol Pharmacol, 15(3- 4), 263-276. https://doi.org/10.1515/JB- CPP.2004.15.3-4.263

Murthy, K.R.K., Zare, M.A. (2002). Scorpion antivenom reverses metabolic, electrocardio- graphic, and  hormonal  disturbances  caused  by the Indian red scorpion Mesobuthus tamu- lus concanesis, Pocock envenomation. J Ven- om Anim Toxins, 8(1), 30-48. http://dx.doi. org/10.1590/S0104-79302002000100004

Murthy, K.R.K. (2000). The scorpion envenoming syndrome: a different perspective. The physi- ological basis of the role of insulin in scorpi- on envenoming. J Venom Anim Toxins, 6(1), 04-51.      http://dx.doi.org/10.1590/S0104-

Murthy, K.R.K., Zare, M.A. (1998). Effect of In- dian red scorpion (Mesobuthus tumulus conca- nesis, Pocock) venom on thyroxine and triiodo- thyronine in experimental acute myocarditis and its reversal by species specific  antiven- om. Indian J Exp Biol, 36(1),16-21. PMID: 9536646

Murthy, K.R.K, Zare, M.A. (2001). The use of an- tivenom reverses hematological and osmotic fragility changes of erythrocytes caused by indi- an red scorpion Mesobuthus tumulus concane- sis Pocock in experimental envenoming. J Ven- om Anim Toxins, 7(1), 113-138. http://dx.doi. org/10.1590/S0104-79302001000100008

Murthy, K.R.K. (2013). Treatment of Scorpion Envenoming Syndrome—Need for Scientific Magnanimity. J Indian Med Assoc, 111, 254- 259.   PMID:24475558

Murthy, K.R.K. (2014a). Cardiac Sarcolemmal Defects in acute myocarditis due to scorpion envenoming syndrome. World J Cardiovas Dis, 4(09), 432-454. http://dx.doi.org/10.4236/ wjcd.2014.49054

Murthy, K.R.K. (2014b). Enzymes and toxins in scorpions of Buthidae family. insulin-glucose administration reverses metabolic, cardiovas- cular, ECG changes and pulmonary edema in scorpion envenoming syndrome. Int J Med Biosciences,  3, 9-25.

 

 

 

Murthy, K.R.K., Haghnazari, L. (1999). The blood levels of glucagon, cortisol and insulin follow- ing the injection of venom by the scorpion (Me- sobuthus tumulus concanesis, Pocock) in dogs. J Venom Anim Toxins, 5(1), 47-55. http://doi. org/10.1590/S0104-79301999000100004

Murthy, K.R.K., Shenoi, R., Vaidyanathan, P., Kelkar, K., Sharma, N., Neeta, B., Rao, S. Me- hta, M.N. (1991) Insulin reverses haemody- namic changes and pulmonary oedema in chil- dren stung by Indian red scorpion Mesobuthus tumulus concanesis, Pocock. Ann Trop Med Parasitol, 85, 651-657. https://doi.org/10.108 0/00034983.1991.11812621

Murthy, K.R.K., Vakil, A.E. and Yeolekar, M.E. (1990) Insulin administration reverses the met- abolic and electrocardiographic changes in- duced by Indian red scorpion (Buthus tamulus) Venom in the Experimental Dogs. Indian Heart J, 48, 35-42. PMID:  2190913

Murthy, K.R.K., Vakil, A.E., Yeolekar, M.E., Va- kil, Y.E. (1988). Reversal of metabolic and electrocardiographic changes induced by Indi- an red scorpion (Buthus tamulus) venom by ad- ministration of insulin, alpha blocker and Sodi- um bicarbonate. Indian J Med Res, 88,450-457. PMID:2904413

Nencioni, A.L.A., Neto, E.B., de Freitas, L.A., Dorce, V.A.C. (2018). Effects of Brazilian scor- pion venoms on the central nervous system. J Venom Anim Toxins Incl Trop Dis, 24(1), 3. http://doi.org/10.1186/s40409-018-0139-x

Petricevich,   V.L.   (2010).   Scorpion   venom  and  the   inflammatory   response.   Media- tors    Inflamm.    2010:    903295.   http://doi.

Rahmani, A.H., Forouzandeh, H., Kalantar, M.,Asad-Masjedi, N., Alavian, Z. and Ka- varizadeh, K. (2015). Epidemiological and clin- ical characteristics of scorpion stings in ahwaz, southwest iran (2006-2010). Int  J  Med Toxi- col Forensic Med, 5(4), 201-216. http://doi. org/10.22037/ijmtfm.v5i4(Autumn).9442

Rahmani, A.H., Jalali, A. (2012). Symptom pat- terns in adult patients stung by scorpions with emphasis on coagulopathy and hemoglubin- uria.  J  Venom  Anim  Toxins  Incl  Trop   Dis,


18(4), 427-431. http://dx.doi.org/10.1590/ S1678-91992012000400011

Razi Jalali, M., Jalali, M.T., Mapar, Z. (2015). Evaluation of plasma cytokine levels in Me- sobuthus eupeus (Scorpionida: Buthidae) scor- pion envenomation in rats treated with poly- valent antivenom. Jundishapur J Health Sci, 7(1),1-5.   http://doi.org/10.5812/jjhs.27159

Razi jalali, M., Jalali, S.M., Najafzadeh Varzi, H., Shokraeian, A.A. (2016). A study on protective effects of polyvalent antivenom and quercetin on hemogram and erythrocyte osmotic fragility changes following Mesobuthus eupeus scorpi- on envenomation in rat. Iran Vet J, 12(1), 30-42 (In Persian with English abstract). http://doi. org/10.22055/ivj.2016.14703

Sadeghian, H. (2003). Transient ophthalmople- gia following envenomation by the scorpion Mesobuthus  eupeus.  Neurology,  60(2),  346-

Sengupta, S., Dhanapal, P., Ravindran, R.D.,  Devi, N. (2009). Cerebral blindness after scorpion sting. J Neuroophthalmol, 29(2), 154-155. https://doi.org/10.1097/WNO.0b 013e31818e40ec

Seyedian, R., Hoseiny, S.M., Kamyab, M., Mans- ury, R., Seyedian , N., Gharibi, S., Zare Mirak- abadi, A. (2014). An in vitro comparative study upon the hemolytic, thrombogenic, coagula- tion parameters and stability properties of the Hemiscorpius lepturus venom.  Arch  Razi Inst, 69(1), 69-76. https://doi.org/10.7508/ ari.2014.01.010

Taibi-Djennah, Z., Laraba-Djebari, F. (2015). Ef- fect of cytokine antibodies in the immunomod- ulation of inflammatory response and metabol- ic disorders induced by scorpion venom. Int Immunopharmacol, 27(1), 122-129. https:// doi.org/10.1016/j.intimp.2015.05.002

Thomas, V.V., George, T., Mishra, A.K., Man- nam, P., Ramya, I. (2017). Lateral medullary syndrome after a scorpion sting. J  Family  Med Prim Care, 6(1), 155-157. https://doi. org/10.4103/2249-4863.214988

Yugandhar, B., Murthy, K.R.K., Sattar, S.A. (1999).

 

 

Insulin administration in severe scorpion en- venoming. J Venom Anim Toxins Incl Trop Dis, 5, 200-219. http://dx.doi.org/10.1590/ S0104-79301999000200007

Zare, M.A., Murthy, K.R.K., Haghnazari, L. (1994). Scorpion venom poisoning in experi- mental animals. Arch Razi Inst,  44(45),67-72.