J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 204 http://jad.tums.ac.ir Published Online: May 27, 2017 Original Article Efficacy of Aloe Vera Cream in the Treatment of Paederus Dermatitis in Mice Ramin Khaghani 1, *Iraj Mirzaii-Dizgah 2, Mostafa Ghasemi 1 1Department of Medical Parasitology, School of Medicine, Aja University of Medical Sciences, Tehran, Iran 2Department of Physiology, School of Medicine, Aja University of Medical Sciences, Tehran, Iran (Received 2 Aug 2014; accepted 7 May 2016) Abstract Background: Dermatitis caused by Paederus beetle involves many people around the world, especially Iran. The symptoms include redness, itching and severe irritation. This study evaluated the effectiveness of the Aloe vera cream on the treatment of dermatitis caused by Paederus beetles. Methods: Forty male 6–8 weeks BALB/C mice were randomly divided into four groups of 10 mice. After removing the mice’s back hair, the backs of mice were marked by a circle with a diameter of 3 mm. The Paederus beetles were collected from Babol in Mazandaran Province, northern Iran and transferred to the animal lab of Aja University of Medical Sciences, Tehran, Iran. The end of abdominal segment Paederus was cut with scissors and hemolymph con- tent was pushed by forceps on the circle. Only hemolymph of one Paederus applied to the back of each mouse. Groups 1, 2, 3 and 4 were treated with the base (vehicle), dexamethasone 0.1%, Aloe vera 0.5% and Aloe vera 2% creams respectively. After 2 days, dermatitis appeared. Then the mentioned creams were applied on the mice once a day. The wound area was measured every day. Dermatitis surface area under curve (AUC) of each mouse was cal- culated for 17 days after induction of dermatitis. Statistical analysis of ANOVA was used. Results: Application of Aloe vera 0.5% and 2% significantly reduced the healing duration and dermatitis area in comparison with the vehicle and dexamethasone cream (P< 0.05). But dexamethasone had no significant effect on the healing of dermatitis as compared to vehicle. Conclusion: Aloe vera may clinically effective in the treatment of Paederus dermatitis. Keywords: Paederus, Dermatitis, Aloe vera Introduction The genus Paederus is related to family Staphyllinidae, order Coleoptae, class Insecta and consists of over 622 species distributed all around the world (Kerdel-Vegas and Goih- man-Yahr 1966, Zargari et al. 2003). Adult rove beetles are between 7–10mm long and 0.5mm wide. These beetles have a black head, a red thorax, upper abdomen, elytral and a lower abdomen (Frank and Kanamitsu 1987, George and Hart 1990). Many species of the rove beetle genus Paederus contain the he- molymph toxin named pederin (Frank and Kanamitsu 1987). This substance causes itch- ing and skin lesions in humans. With their hemolymph penetrated into the skin, these re - sults in an affliction called dermatitis line- aris. The severe cytotoxic effects of pederin are based on blocking of protein synthesis and inhibition of mitosis in eukaryotic cells (Brega et al. 1968). This dermatitis is more frequent on the uncovered parts of the body like face, neck and hands. The acute lesions become crusted within a few days and heal completely in about 2 to 3 weeks and may leave a hyper- pigmentation (Kerdel-Vegas and Goihman- Yahr 1966, Gelmetti and Grimalt 1993). Recently, the production of pederin is depended on the activities of an endosymbi- ont (Pseudomonas species) within Paederus. *Corresponding author: Dr Iraj Mirzaii-Dizgah, Email: emirzaii@razi.tums.ac.ir J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 205 http://jad.tums.ac.ir Published Online: May 27, 2017 The manufacture of pederin is mostly con- fined to adult females. Larva and males store pederin in very small quantities compared to females (Piel 2002). Aloe vera (family: Liliaceae) has been used in traditional medicine since ancient times. It is one of the most useful herbs in the world and the medicinal part is the leaves. Aloe vera leaves contain various es- sential nutrients for the body, such as amino acids, B family vitamins, etc. It also has pharmacological effects like antioxidant, wound healing, antifungal, antibacterial and immunomodulatory effects (Rodriguez-Bigas et al. 1988, West and Zhu 2003, Hu et al. 2005, Tai-Nin Chow et al. 2005, Olaleye et al. 2005, Rosca-Casian et al. 2007, Habeeb et al. 2007, Maenthaisong et al. 2007). Aloe vera also has anti-inflammatory prop- erties (Reynolds and Dweck 1999, Vogler and Ernst 1999). Compounds existing in the inner gel including salicylates, magnesium lactate, bradykinin, thromboxane inhibitors, sterols and a beta linked acetyl mannan (acemannan) have demonstrated anti-inflammatory activity (Vázquez et al. 1996, Talmadge et al. 2004). The mucilaginous polysaccharides in the pulp of Aloe vera leaf are the major ingredient that induces regeneration. Emodin - as a derivative of anthraquinones produced by pericyclic cells on the surface of Aloe leaves- can also pro- mote the healing of rats’ excisional wounds by stimulating regeneration of cells (Eshun et al. 2004, Tang et al. 2007). These researches support the claim that the healing properties of Aloe are essentially result of the synergis- tic mode of action of many bioactive ingre- dients of Aloe leaves (Dagne et al. 2000). The purpose of the present study was to de- termine the efficacy of Aloe vera in the treat- ment of dermatitis caused by Paederus beetles. Materials and Methods Animals Forty male BALB/C mice (6–8 weeks old) were obtained from Pasteur Institute of Iran. Animals had free access to food and water and were kept at room temperature and with an artificial light cycle of 12-h light and 12-h dark. All animal care and procedures and han- dling were performed under supervision of Animal Care and Use Committee of the Aja University of Medical Sciences. Chemicals Dexamethasone acetate (CAS Number: 464-92-6) was purchased from Chemos GmbH (Regenstauf, Germany). All other chemicals used in this study were of analytical grade and obtained from Merck (Darmstadt, Germany). Aloe vera The pure spray-dried Aloe vera powder was purchased from Anamis Company (Al- borz, Iran). This product consists of the inner gel of the leaves of Aloe vera. Paederus Collection and Identification The Paederus beetles were collected from Babol in Mazandaran Province of Iran and transferred to the animal lab of Aja Univer- sity of Medical Sciences, Tehran, Iran. Iden- tification was made by specialists and by using the key and illustrations of Coiffait (1982). Paederus beetles were identified as Paederus fuscipes that together with Pa. kalalovae are the predominant species along the southern shores of the Caspian Sea (Nikbakhtzadeh and Tirgari 2008). Dermatitis induction and treatments Forty male 6–8 weeks BALB/C mice were randomly divided into four groups (10 mice in each group). Each mouse was shaved on the back (2cm× 2cm) and the skin was gen- tly wiped with distilled water. The backs of mice were marked by a circle with a diameter of 3 millimeters. The end of abdominal seg- ment of adult female Paederus beetles was cut with scissors and hemolymph content was J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 206 http://jad.tums.ac.ir Published Online: May 27, 2017 push by forceps on the circle. Only hemo- lymph of one Paederus applied to the back of each mouse. After 48 h the dermatitis appeared clearly (Fig. 1) and its surface area was measured and recorded as per square millimeter (mm2). Groups 1, 2, 3 and 4 were treated with the base cream (as vehicle group), dexamethasone 0.1 % cream, Aloe vera 0.5% cream and Al- oe vera 2% cream respectively. Drug doses had been shown in a previous study to be adequate to elicit a response (Babaee et al. 2012) Preparation of the creams Dexamethasone and Aloe vera powder were formulated in an aqueous cream containing polysorbate 80 (5w/v), Isopropyl myristate, stearyl alcohol (20v/v), liquid white paraffin (15v/v), purified water (100ml), methylpara- ben (0.18w/w) and propylparaben (0.02w/w). Preparation process carried out under condi- tions in which dexamethasone and Aloe vera powder were not heated. The creams con- tained Aloe vera powder, dexamethasone cream and base cream were prepared accord- ing to similar protocol, under the same con- ditions and with the same chemicals. Dermatitis surface area measurement To determine the surface areas of derma- titises, the mice were observed every day. A photograph was taken of each mouse’s der- matitis with a single lens, 14.1MP Digital Camera (SX30 IS, Canon, Japan). For cali- brating the magnification of photos, cam- era’s lens was held at a distance of 20cm from the dermatitis and a caliper was placed at the level of mice’s back. The wound area was measured per mm2 every day by analysis of images using Image J (Image Processing and Analysis in Java, US National Institutes of Health) and Adobe Photoshop CS5 soft- ware. Dermatitis surface area under curve (AUC) of each mouse was calculated for 17d after induction of dermatitis (dermatitis sur- face in mm2 × time between observations in day). Statistical analysis Values are expressed as Mean± SEM. One- way analysis of variance, ANOVA (post hoc Tukey test) was used to determine significant differences among groups and P< 0.05 was considered a significant difference. Statisti- cal analysis performed using SPSS software (version 15, SPSS Inc., Chicago, IL, USA). Results In the second day after dermatitis induction, there was no significant difference in the dermatitis surface area values among groups. The mean ( SEM) dermatitis surface areas were 19.610.43mm2, 19.890.43mm2, 19.34 0.46mm2 and 19.50.51mm2 for group 1 to 4, respectively. A one-way ANOVA indicated that the means AUC of the dermatitis surface was significantly different among groups (P< 0.05, Fig. 2). Post-hoc analysis showed that Aloe vera treatment dose dependently de- creased the AUC of the dermatitis surface as compared to vehicle and dexamethasone treat- ments (P< 0.05). There was no significant dif- ference between vehicle and dexamethasone treatments and also between two doses of Aloe vera creams (P> 0.05). Aloe vera creams (0.5% and 2%) signifi- cantly reduced the healing period of derma- titis in comparison with dexamethasone and base creams (groups 1 and 2) (P< 0.05, Fig. 3). But there was no significant difference between two Aloe vera creams. Dexame- thasone cream and vehicle had no significant difference in reduction of healing duration of dermatitis (Fig. 3). Healing time ranged from 14 to 17 days in vehicle group, 15 to 17 days in dexamethasone 1% treated group, 13 to 16 days in Aloe vera 0.5% treated group and 12 to 14 days in Aloe vera 2% treated group. J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 207 http://jad.tums.ac.ir Published Online: May 27, 2017 Fig. 1. Dermatitis induced by Paederus hemolymph on the back of mouse. *# *# 0 200 1 A U C (m m 2 .d ay ) Dexamethasone 0.1% Vehicle Aloe vera 0.5% Aloe vera 2% Fig. 2. Effect of treatment with base cream (vehicle), dexamethasone 1% or Aloe vera cream 0.5 and 2% on dermatitis surface changes for 17 day observations (as indicated by area under curve (AUC)) following exposure to Paederus’ hemolymph in BALB/C mice. Data are expressed as mean  SEM. *and # different from vehicle and dexamethasone 0.1% treated, re- spectively, P< 0.05. *# *# 0 3 6 9 12 15 18 1 H ea lin g pe rio d (d ay ) Dexamethasone 0.1% Vehicle Aloe vera 0.5% Aloe vera 2% Fig. 3. Effect of treatment with base cream (vehicle), dexamethasone 1% or Aloe vera cream 0.5 and 2% on the days passed for completing healing induced by Paederus’ hemolymph in BALB/C mice. Data are expressed as mean  SEM. *and # different from ve- hicle treated (vehicle) and dexamethasone 0.1 % treated, respectively, P< 0.05. Discussion Dermatitis caused by Paederus beetle in- volves many people around the world, espe- cially in Iran. The symptoms include redness, itching and severe irritation. In this study the efficacy of the Aloe vera cream on the treat- ment of dermatitis caused by Paederus bee- tles was investigated. Aloe vera creams (0.5 and 2 percent) decreased the healing period of dermatitis and AUC of the dermatitis sur- face. Thus it seems that Aloe vera creams (0.5 and 2 percent) have capability to promote dermatitis healing more effectively than dex- amethasone and base creams. Up to now, numerous studies have been performed on the effects of Aloe vera. Alt- hough there are many studies that have shown the efficacy of different Aloe vera prepara- tion on various dermatologic disorders such as fungal and bacterial infections, incision and burn wounds, skin inflammations and many others (Rodriguez-Bigas et al. 1988, Vázquez et al. 1996, Reynolds and Dweck 1999, Vogler and Ernst 1999, Yagi et al. 2002, Rosca-Ca- sian et al. 2007, Maenthaisong et al. 2007), but its effect on dermatitis linearis caused by pederin existing in Paederus’ hemolymph was unknown. “Aloe vera mouthwash was effective in healing of the wound and reducing the in- flammation of the mucous membrane of the mouth” (Mansour et al. 2013). The topical ap- plication of Aloe vera improves the total qual- ity of life score in patients with oral lichen planus (Salazar-Sánchez et al. 2010). Further- more, its gel has been effective in healing of the first and second degree burning wounds without any side effects (Maenthaisong et al. 2007) and also in cesarean wound healing (Tarameshloo et al. 2012, Molazem et al. 2014). Aloe vera 2% oral gel is effective in decreas- ing the recurrent aphthous stomatitis patients' pain score and wound size and decreases the aphthous wound healing period too (Babaee J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 208 http://jad.tums.ac.ir Published Online: May 27, 2017 et al. 2012). Aloe vera reduces inflammatory cytokines (such as IL-18), INF-α neutrophil chemoattractant, and malondialdehyde as in- dicator of oxidative stress (Werawatganon et al. 2014). Many of the biological activities of Aloe vera such as immunostimulation, anti-inflam- matory effects, wound healing, promotion of radiation damage repair, anti-bacterial, anti- viral, anti-fungal, anti-diabetic and anti-neo- plastic activities, and stimulation of hemato- poiesis and anti-oxidant effects have been attributed to the polysaccharides contained in the gel of the leaves. These biological activi- ties should be assigned to a synergistic ac- tion of the compounds contained therein ra- ther than a single chemical substance (Ham- man 2008). Results of our study showed that there was no significant difference between dexa- methasone cream and vehicle in reducing the dermatitis healing duration. This is in agree- ment with Qadir et al. report (2006) that com- bination of topical steroids and oral antibiot- ics have better effect than topical steroids alone on treatment of Paederus dermatitis in Sierra Leone patients. Cortisone or antihis- tamine preparations have no beneficial effect (Deneys and Zumpt 1963). As dexamethasone is 25 times more potent than cortisol in its glucocorticoid effect, it seems that due to anti-inflammatory and immunosuppressant ef- fects of glucocorticoids, without antibiotic medication, these medications can be even harmful by favoring secondary infections. Conclusion Aloe vera seems to be clinically effective in the treatment of Paederus dermatitis. It may be considered as an alternative or sup- plementary medicine for patients with this disease. Further investigation on human and prolonged follow-up period is proposed in order to confirm the efficacy of Aloe vera in the treatment of Paederus dermatitis in human. Acknowledgments We gratefully acknowledge grant of this study by Aja University of Medical Sciences. We also thank Drs Banafsheh Dormanesh and Mehdi Mirqazanfari for their supports. There was no conflict of interest for this study. References Agarry OO, Olaleye MT, Bello-Michael CO (2005) Comparative antimicrobial ac- tivities of Aloe vera gel and leaf. Afr J Biotechnol. 4(12): 1413–1414. Babaee N, Zabihi E, Mohseni S, Moghadam- nia AA (2012) Evaluation of the ther- apeutic effects of Aloe vera gel on mi- nor recurrent aphthous stomatitis. Dent Res J (Isfahan). 9(4): 381–385. Brega A, Falaschi A, Carli L De, Mario P (1968) Studies on the mechanism of action of pederine. J Cell Biol. 36(3): 485–496. Coiffait H (1982) Coleopteres Staphylinidae de la region Palearctique Occidentale. Sous famille Paederinae, Tribu Paederini (Paederi, Lathrobii). Nouv Rev Ento- mol. 7: 1–440. Dagne E, Bisrat D, Viljoen A, Van Wyk BE (2000) Chemistry of Aloe species. Curr Org Chem. 4(10): 1055–1078. Eshun K, He Q (2004) Aloe vera: a valuable ingredient for the food, pharmaceutical and cosmetic industries-a review. Crit Rev Food Sci Nutr. 44(2): 91–96. Frank J H and Kanamitsu K (1987) Paede- rus. sensu lato (Coleoptera: Staphylin- idae): natural history and medical im- portance. J Med Entomol. 24(2): 155– 191. Qadir SN, Raza N, Rahman SB (2006) Pae- derus dermatitis in Sierra Leone. Der- matol Online J. 12(7): 9. Gelmetti C, Grimalt R (1993) Paederus der- matitis: An easy diagnosable but mis- J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 209 http://jad.tums.ac.ir Published Online: May 27, 2017 diagnosed eruption. Eur J Pediatr. 152 (1): 6–8. George AO, Hart PD (1990) Outbreak of Paederus Dermatitis in Southern Nige- ria. Int J Dermatol. 29(7): 500–501. Habeeb F, Shakir E, Bradbury F, Cameron P, Taravati MR, Drummond AJ, Gray AI, Ferro VA (2007) Screening meth- ods used to determine the anti-micro- bial properties of Aloe vera inner gel. Methods. 42(4): 315–320. Hamman JH (2008) Composition and appli- cations of Aloe vera leaf gel. Mole- cules. 13(8): 1599–1616. Hu Q, Hu Y, Xu J (2005) Free radical-scav- enging activity of Aloe vera (Aloe bar- badensis Miller) extracts by supercriti- cal carbon dioxide extraction. Food Chem. 91(1): 85–90. Deneys JB, Zumpt F (1963) Rove beetel dermatitis in south West Africa. S Afr Med J. 37: 1284–1285. Kerdel-Vegas F, Goihman-Yahr M (1966) Paederus dermatitis. Arch Dermatol. 94(2): 175–185. Maenthaisong R, Chaiyakunapruk N, Ni- runtraporn S, Kongkaew C (2007) The efficacy of Aloe vera used for burn wound healing: A systematic review. Burns. 33(6): 713–718. Mansour G, Ouda S, Shaker A, Abdallah HM (2014) Clinical efficacy of new Aloe vera- and myrrh-based oral mu- coadhesive gels in the management of minor recurrent aphthous stomatitis: a randomized, double-blind, vehicle-con- trolled study. J Oral Pathol Med. 43(6): 405–409. Molazem Z, Mohseni F, Younesi M, Keshavar- zi S (2014) Aloe vera gel and cesarean wound healing, a randomized controlled clinical trial. Glob J Health Sci. 7(1): 203–209. Nikbakhtzadeh M, Tirgari S (2008) Medi- cally important beetles (insecta: cole- optera) of Iran. J Venom Anim Toxins. 14(4): 597–618. Piel J (2002) A polyketide synthase-peptide synthetase gene cluster from an un- cultured bacterial symbiont of Paede- rus beetles. Proc Natl Acad Sci USA. 99(22): 14002–14007. Reynolds T, Dweck AC (1999) Aloe vera leaf gel: a review update. J Ethnophar- macol. 68(1–3): 3–37. Rodriguez-Bigas M, Cruz NI, Suarez A (1988) Comparative evaluation of Aloe vera in the management of burn wounds in guinea pigs. Plast Reconstr Surg. 81 (3): 386–389. Rosca-Casian O, Parvu M, Vlase L, Tamas M (2007) Antifungal activity of Aloe vera leaves. Fitoterapia. 78(3): 219–222. Salazar-Sánchez N, López-Jornet P, Camacho- Alonso F, Sánchez-Siles M (2010) Ef- ficacy of topical Aloe vera in patients with oral lichen planus: a randomized double-blind study. Oral Pathol Med. 39(10): 735–740. Tai-Nin Chow J, Williamson DA, Yates KM, Goux WJ (2005) Chemical character- ization of the immunomodulating pol- ysaccharide of Aloe vera L. Carbohydr Res. 340(6): 1131–1142. Talmadge J, Chavez J, Jacobs L, Munger C, Chinnah T, Chow JT, Williamson D, Yates K (2004) Fractionation of Aloe vera L. inner gel, purification and mo- lecular profiling of activity. Int Im- munopharmacol. 4(14): 1757–1773. Tang T, Yin L, Yang J, Shan G (2007) Emodin, an anthraquinone derivative from Rheum officinale Baill, enhances cutaneous wound healing in rats. Eur J Pharmacol. 567(3): 177–185. Tarameshloo M, Norouzian M, Zarein-Dolab S, Dadpay M, Mohsenifar J, Gazor R (2012) Aloe vera gel and thyroid hor- mone cream may improve wound heal- ing in Wistar rats. Anat Cell Biol. 45 (3): 170–177. Vázquez B, Avila G, Segura D, Escalante B J Arthropod-Borne Dis, June 2017, 11(2): 204–210 R Khaghani et al.: Efficacy of Aloe Vera … 210 http://jad.tums.ac.ir Published Online: May 27, 2017 (1996) Antiinflammatory activity of ex- tracts from Aloe vera gel. J Ethnophar- macol. 55(1): 69–75. Vogler BK, Ernst E (1999) Aloe vera: a sys- tematic review of its clinical effective- ness. Br J Gen Pract. 49(447): 823–828. Werawatganon D, Rakananurak N, Sallapant S, Prueksapanich P, Somanawat K, Klaikeaw N, Rerknimitr R (2014) Aloe vera attenuated gastric injury on in- domethacin-induced gastropathy in rats. World J Gastroenterol. 20(48): 18330– 18337. West DP, Zhu YF (2003) Evaluation of Aloe vera gel gloves in the treatment of dry skin associated with occupational ex- posure. Am J Infect Control. 31(1): 40–42. Yagi A, Kabash A, Okamura N, Haraguchi H, Moustafa SM, Khalifa TI (2002) Antioxidant free radical scavenging and anti-inflammatory effects of al- oesin derivatives in Aloe vera. Planta Med. 68(11): 957–960. Zargari O, Kimyai-Asadi A, Fathalikhani F, Panahi M (2003) Paederus dermatitis in northern Iran: a report of 156 cases. Int J Dermatol. 42(8): 608–612.