In-Vitro and In-Vivo Antibacterial Effect of Croton Lobatus Linnaeus L. on Two Days Croton Lobatus L

Available online at www.banglajol.info Bangladesh J. Sci. Ind. Res. 54(2), 139-146, 2019

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro In-vitro and in-vivo antibacterial effect of Croton lobatus Linnaeus L. on two days Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des post surgical wounds in rats areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative method was used to estimate total phenol contents of the alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 1 1 1 2 3 1 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. M. A. Kilani *, A. Z. Hassan , S. T. Fadason , A. M. Obalowu , A. Aliyu and H. M. Badmus-Kilani (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family Examinations of Antimicrobial activity of the C. lobatus Sofowara A (1993), Medicinal plants and Traditional 1 parasitic infection, treatment for menstrual cycle disorders, Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Conclusion Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Euphorbiaceae In: The Useful Plants of West Department of Veterinary Surgery and Radiology, Ahmadu Bello University, P. M. B. 1096, Zaria, Kaduna State, Nigeria Preparation of crude extract Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, 2 antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Tropical Africa (2), Burkill HM, Royal Botanic University of Ilorin Veterinary Teaching Hospital, Ilorin, Kwara State, Nigeria Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. 3 paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Gardens, Kew (K), Department of Veterinary Surgery and Radiology, University of Ilorin, Kwara State, Nigeria Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): 4 temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem Ministry of Health, General Hospital Hunkuyi, Kaduna State, Nigeria (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 Abstract infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). In vitro antimicrobial assay therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences Van-Burden TP and Robinson T (1981), The biochemistry of Phytochemical constituents of Croton lobatus L. (C. lobatus) water extracts and quantitative analysis Croton lobatus L. has recently been reported by traditional retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: involve microbial infections. Further research is required 13(2): 001 alkaloids. 2nd Ed. Springer, Heidelberg, Newyork. were carried out following standard procedures. The antibacterial activity against Staphylococcus medicine practitioner in Nigeria as having a good therapeutic Streptococcus Spp; Pseudomonas aeruginosa; Proteus 203-208. DOI:org/10.4314/gjpas.v8i2.16033 Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) on this plant. aureus (ATCC 33591); Streptococcus Spp; Pseudomonas aeruginosa (ATCC 9028); Proteus index on chronic wounds by applying its fresh leaves or dried vulgaris; E. coli and Salmonella Spp at the zone of Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata was adopted. The prepared C. lobatus water extract was Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M vulgaris; Escherichia coli (ATCC 43895); and Salmonella Spp (ATCC 4932) was carried out at the powder on wound surface while the leaf-decoction is used to inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth Jiwan JLH, Llabrès G, Pauw-Gillet MC and (Euphorbiaceae), a new record for India, Rheedea Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Acknowledgment and Shuaib B (2012), Documentation of Wound concentration of 0.5g/mL, 0.05 g/mL and 0.00 5g/mL of water. In vivo antimicrobial assay was aid development of fetus. Hence, it is necessary to screen for of Proteus vulgaris at 34mm while C. lobatus 0.005 Quetin-Leclercq J (2004), Effects of Croton 22(2): 131-132. alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Healing Plants in Lagos-Nigeria: Inhibition of Lipid carried out by creating four wounds of 0.5 by 0.5 cm on dorsal surface of a male albino rat under the phytoconstituents of C. lobatus and evaluate its inhibited the growth of Staphylococcus aureus at 18mm rhamnifolioides Essential Oil on Aedes, glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the Peroxidation as In-vivoPrognostic Biomarkers of anesthesia. The wounds were left for 48 hrs, after which they were accessed and samples were antimicrobial activities against common microorganisms that (Fig. 2). CFU counted on nutrient agar reduced drastically Phytochemistry 65: 1165. Global Biodiversity Information Facility, GBIF (2016), (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Activity, Annals of Biological Research 3(4): collected for culture, identification and colony forming unit counts (CFU). Respective treatment after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 using dried C. lobatus, C. lobatus (water extract), Physiological saline solution and Cicatrin powder 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum was carried out and samples were collected at day one, three, five and seven after initiation of earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and treatments for CFU counts on nutrient and MacConkey agar. The phytochemical studies revealed Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of that C. lobatus contains carbohydrates, glycoside, saponins, steroids, triterpenes, flavonoids, cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus alkaloids and tannins. Croton lobatus L. showed a dose dependent activity against micro organisms with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the aureus and Escherichia coli, Journal of Health with C. lobatus 0.5 inhibited the growth of most bacteria at the zone of inhibition ≤ 21mm. This was retinoid-X receptor, Archives of Biochemistry and 0.005 g/mL extracts were filled into each of the wells. The carried out after 48 hrs of wound creation (Table I) and Ivory Coast, Travaux Ed Qocumentes de I’orstom Research 23(30): 153-156. also supported by in vivo antimicrobial assay. Secondary metabolite tannins, triterpenoids, Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. plates were incubated at 37°C for 24 hrs. The diameters of the samples were collected at day one, three, five and seven after Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. flavonoids, crotonic acids and saponin were responsible for its antimicrobial activity against the 2003.09.034 Received: 29 March 2018 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes tested microorganisms thereby supporting its usage by the traditional medicine practitioner in inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial Revised: 25 Novembe 2018 Nigeria to treat chronic wounds. 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), Accepted: 03 December 2018 In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. Keywords: Phytochemical screening; Antibacterial; Croton lobatus L.; Surgical wounds; Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 Astraea lobata The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of Introduction humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 0378-8741 (94)90041-8 major languages which are: Gaásàyaá (Hausa); (prepared from vital feed grower mash) and adequate water Aké AL (1975), Contribution aux études ethnobotaniques of Science and Technology 5(1): 1936- 1940. One thousand three hundred and thirty species of trees, Phytochemical Studies Chabert P, Attioua B, Weniger B and Brouillard R (2006), Òkwè-one a kind of bean (Igbo); Àjẹìofòlé or ẹÌru was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), shrubs and herbs belonging to Croton were distributed in Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton (Yoruba). The old name was Astraea lobata Klotzsch Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at the Northern and Southern Hemispheres (Block et al. permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 (Global Biodiversity Information Facility, 2016; constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton 2004). Majority of which were popularly used for various was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT Malpighiales of North America Update, 2011). Croton &p=Croton+macrostachyus#Synonyms. Accesed on purposes due to their medicinal qualities in different experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T lobatus L. is native of Caribbean and American but (R) 07.012.2015 at 22:00GMT. continents (Yibralign, 2007). Croton lobatus L. is Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), introduced in different part of African countries commonly known as Rusffoil; Croton; Lobed croton and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (Gaikwad et al., 2012; Malpighiales of North America (R) Salatino A, Salatino MLF and Negri G (2007), Traditional (Malpighiales of North America Update, 2011). In whole body of the animal was cleaned with diluted Purit . Its This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into ent-Beyer-15-en-18-oic acid, Pharmaceutical Update, 2011). dorsal region was surgically prepared for aseptic surgery and uses, Chemistry and Pharmacology of Croton species Nigeria, Burkill reported that the plant is known in three glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 *Corresponding author e-mail: [email protected] 140 In-vitro and in-vivo antibacterial effect of Croton lobatus Linnaeus L. 54(2) 2019

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of method was used to estimate total phenol contents of the feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family parasitic infection, treatment for menstrual cycle disorders, Examinations of Antimicrobial activity of the C. lobatus Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West Preparation of crude extract Conclusion Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical Table I. Concentration of C. lobatus water extracts and standard antibiotics used for both in vitro and in vivo treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), antimicrobial studies counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of Antibiotic disk cartridges (Treatment) cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus Code Disk Potency (treatment dose) with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health 1 C. lobatus CL – P (A) Dried Paste (Liberal) carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. 2 C. lobatus CL – 0.5 (B) 0.5 g/mL of water (2 drops) samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 3 C. lobatus CL – 0.05 0.05 g/mL of water growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes 4 C. lobatus CL – 0.005 0.00 5g/mL of water inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 5 Ceftriaxone CRO 30 30 μg 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. 6 Erythromycin E 15 15 μg Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 7 Sulphamethoxazole STX 25 25 μg The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia 8 Cloxacillin OB 5 5 μg bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. 9 Oxacillin OX 1 1 μg State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 10 Ceftazidime CAZ 10 10 μg 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water of Science and Technology 5(1): 1936- 1940. 11 Gentamycin CN 10 10 μg Phytochemical Studies Aké AL (1975), Contribution aux études ethnobotaniques Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), 12 Amoxycillin AML 25 25 μg Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ 15 Physiological saline solution (PSS) C 2 drops Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton 16 Cicatrin powder D Liberal was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on KEY: CL – P (C. lobatus Paste), CL – 0.5 (C. lobatus: 0.5g/mL of water), CL – 0.05 (C. lobatus: 0.05g/mL of water), CL – 0.005 (C. experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. lobatus: 0.005g/mL of water), CRO 30 (Ceftriaxone), E 15 (Erythromycin), STX 25 (Sulphamethoxazole), OB 5 (Cloxacillin), OX 1 Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), (Oxacillin), CAZ 10 (Ceftazidime), CN 10 (Gentamycin), AML 25 (Amoxycillin). and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into ent-Beyer-15-en-18-oic acid, Pharmaceutical dorsal region was surgically prepared for aseptic surgery and uses, Chemistry and Pharmacology of Croton species glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 Kilani, Hassan, Fadason, Obalowu, Aliyu and Badmus-Kilani 141

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative method was used to estimate total phenol contents of the alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family parasitic infection, treatment for menstrual cycle disorders, Examinations of Antimicrobial activity of the C. lobatus Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West Preparation of crude extract Conclusion Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay Fig. 1. Surgical creation of four wounds on the dorsal retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd surface of the rat. Wound treated with C. lobatus paste (A), Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) C. lobatus water extract (B), normal saline (C) and Cicatrin vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, powder (D) inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water of Science and Technology 5(1): 1936- 1940. Phytochemical Studies Aké AL (1975), Contribution aux études ethnobotaniques Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its ent-Beyer-15-en-18-oic acid, Pharmaceutical This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into uses, Chemistry and Pharmacology of Croton species dorsal region was surgically prepared for aseptic surgery and glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 142 In-vitro and in-vivo antibacterial effect of Croton lobatus Linnaeus L. 54(2) 2019

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey Table II. Phytochemical constituents of C. lobatus to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it hemiargyreus, Phytochemistry 47: 1445. repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically S/N Chemical Qualitative Quantitative al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et Public Health and Clinical Nutrition, Faculty of Health from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local Constituents/ test test (%) activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Test reagent Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of 1 Carbohydrates feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis mouth, or a bark-decoction by enema is given as a purgative method was used to estimate total phenol contents of the alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, to cure respiratory diseases, African Journal of j.jep.2003.10.002 (Burkill, 1985). The plant is used generally as healing leaf Ahmadu Bello University, Zaria, with voucher specimen plant. Molisch’s test + Similar studies have revealed the presence of triterpenes (Euphorbiaceae), Ph.D. Dissertation, University of London, pp 542-578. WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. Fehling’s test + al., 1997; Salatino et al., 2007). medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family Examinations of Antimicrobial activity of the C. lobatus 2 Cardiac glycoside Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West parasitic infection, treatment for menstrual cycle disorders, Kella-killiani test + Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Conclusion Preparation of crude extract Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic antidotes (venomous stings and bites), treatment for Kedde’s test + Amaral and Barnes, 1998; Milanowski et al., 2002; Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), paralysis, epilepsy, convulsions, spasm and topically to treat Leave of C. lobatus was harvested and air dried at normal 3 Free Anthraquinones Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and of Current Research and Academic Review 3(6): International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. temperature. The dried powder of the plant was extracted Borntrager’s test - (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, Ecology 313: 07–308. laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); 477–489. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water with distilled for 24 h. The extract was filtered through 4 Combined Anthracene steroids, triterpenes, flavonoids and tannins, which are Proteus vulgaris (isolate identified from bacteria zoonosis Antibacterial activities in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Whatmann filter paper No 1 and it was thereafter, centrifuged Modified Borntrager’s test - responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); 5 Saponins 3.04 Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant at 5000G for 15 min. The supernatant were further filtered explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and Salmonella Spp (ATCC 4932). Frothing test + Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, Hemolysis test + micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay 6 Steroids and Triterpenes retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd Leiberman-Burchards test + Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) 7 Flavonoids 8.60 vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, Shinoda test + inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and Sodium hydroxide test + of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of 8 Tannins 3.80 Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Lead sub-acetate test + Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Ferric chloride test + Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the 9. Alkaloids 5.22 Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical Mayer’s test + treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Dragendorff’s test + Ongsakul M, Jundarat A and Rojanaworant C (2009), 10. Phenols 3.40 counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Key: +: Present, - : Absent Extracts of six medicinal plants against Staphylococcus with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The 40 Invitro antimicrobial activity of Croton lobatus. Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and 35 potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes inhibition was compared with standard antibiotics (Table I). MacConkey agar. 30 Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), Data analysis 25 In vivo antimicrobial assay Staphylococcus aureus Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. 20 Streptococcuss spp Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was Pseudomonas aeruginosa (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. 15 Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu Proteus vulgaris membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing 10 Escherichia coli lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, Zone of inhibitions (mm) DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion Salmonella DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 5 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water Aké AL (1975), Contribution aux études ethnobotaniques of Science and Technology 5(1): 1936- 1940. Phytochemical Studies 0 Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Antimicrobial agents Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Fig. 2. The clear zone of inhibitions (mm) of various concentration of C. lobatus plant extract compared with standard Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the antibiotic drugs (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. KEY : CL – P (C. lobatus Paste), CL – 0.5 (C. lobatus: 0.5g/mL of water), CL – 0.05 (C. lobatus: 0.05g/mL of water), CL – 0.005 (C. lobatus: Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its ent-Beyer-15-en-18-oic acid, Pharmaceutical This study revealed that C. lobatus contains carbohydrates, 0.005g/mL of water), CRO 30 (Ceftriaxone), E 15 (Erythromycin), STX 25 (Sulphamethoxazole), OB 5 (Cloxacillin), OX 1 (Oxacillin), biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into uses, Chemistry and Pharmacology of Croton species dorsal region was surgically prepared for aseptic surgery and glycoside, saponins, steroids, triterpenes, flavonoids and CAZ 10 (Ceftazidime), CN 10 (Gentamycin), AML 25 (Amoxycillin) Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 Kilani, Hassan, Fadason, Obalowu, Aliyu and Badmus-Kilani 143

metabolites such as carbohydrates, glycoside, saponins, temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); 250 Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water A 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis steroids, triterpenes, flavonoids and tannins, which are in Wound Healing Studies of Plants, Evidence-Based Antibacterial activities N A bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal ( 200 Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. while Bouquet and Debray, (1974) reported the whole plant t Farnsworth (1969), Croton lobatus Linn. Linn. family Thesis. Addis Ababa University, Addis Ababa. n explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against 150 B and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 u Euphorbiaceae In: The Useful Plants of West infusion to be used as a topical bath to treat skin diseases. identification of alkaloids from C. lobatus, through Millipore filter before storage at 4° C as described by micro organisms. Croton lobatus paste and C. lobatus 0.5 o Nigeria and various part of the world as alternative homeostatic plants in Edo and Delta states of Nigeria,

c Tropical Africa (2), Burkill HM, Royal Botanic

C (Prashanthi et al., 2012). In vitro antimicrobial assay 100 therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of retarded the growth of Staphylococcus aureus; y Croton lobatus L. has recently been reported by traditional Gardens, Kew, UK. nd n involve microbial infections. Further research is required 13(2): 001 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Streptococcus Spp; Pseudomonas aeruginosa; Proteus o D

l 50 Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) on this plant. vulgaris; E. coli and Salmonella Spp at the zone of o Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth C 0 (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Acknowledgment Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Day 2 Day 3 Day 5 Day 7 Day 9 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Number of days after the surgery Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical Fig. 3. Colony forming units (CFU) counted on nutrient agar (NA) inoculated with swaps collected from wounds treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were treated with different agents: A (C. lobatus paste); B (C. lobatus water extract); C (Physiological normal Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective saline) and D (Cicatrin powder) Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), unsaturated fatty acids as natural ligands for the Extracts of six medicinal plants against Staphylococcus Physiological saline solution (PSS), Cicatrin powder was with C. lobatus paste while persistent high number of CFU medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and 5 References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and Invivo antimicrobial activity MAC x 10 (Croton lobatus ) 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after ) Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial plates were incubated at 37°C for 24 hrs. The diameters of the 5 350 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and 0 potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes 1

MacConkey agar. Burkill HM (1985), Croton lobatus Linn, family RA (1997), Chemical composition and antimicrobial X 300 inhibition was compared with standard antibiotics (Table I). Zygotritonia crocea, Phytotherapy Research 13(6): Harborne JB (1973), Phytochemcial methods, London,

C Euphorbiaceae In: The Useful Plants of West Tropical activity of Croton urucurana Baillon (Euphorbiaceae), 250 494-7. Chapman and Hall Ltd., pp 49-188. In vivo antimicrobial assay Data analysis A Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6.

M Aboagye FA, Sam GH, Massiot GL and Julocrotine CA DOI:org/10.1016/S0378-8741(97)00039-1 ( 200 A Irobi ON, Moo-Young M, Anderson WA and Daramola SO The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was t (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia selected from an inbred colony obtained from Jos, Plateu bars representing standard error of means. n 150 B Y (1991), Effects of Sangre de Drago from Croton Prashanthi R, Mohan N and Siva GV (2012), Wound healing u membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of

State. It was housed in a steal cage under normal temperature, o lechleri Muell, Phytochemistry 30: 2033. property of aqueous extract of seed and outer layer of c DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ humidity and light. The rat was fed with commercial rat feed Results and discussion 100 C DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal y 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water n of Science and Technology 5(1): 1936- 1940. o 50 D Aké AL (1975), Contribution aux études ethnobotaniques

Phytochemical Studies l was provided. Ethical permission was granted by the Ahmadu Chabert P, Attioua B, Weniger B and Brouillard R (2006), o et ﬂoristiques au Togo, Collection “Médecine Croton lobatus Linn., an African Medicinal plant: Malpighiales of North America Update, database (2011),

C 0 PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Day 2 Day 3 Day 5 Day 7 Day 9 Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT Number of days after the surgery &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Fig. 4. Colony forming units (CFU) counted on MacConkey (MAC) agar inoculated with swaps collected from wounds Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. whole body of the animal was cleaned with diluted Purit(R). Its Salatino A, Salatino MLF and Negri G (2007), Traditional This study revealed that C. lobatus contains carbohydrates, treated with different agents: A (C. lobatus paste); B (C. lobatus water extract); C (Physiological normal saline) and D biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into ent-Beyer-15-en-18-oic acid, Pharmaceutical dorsal region was surgically prepared for aseptic surgery and uses, Chemistry and Pharmacology of Croton species glycoside, saponins, steroids, triterpenes, flavonoids and (Cicatrin powder) Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 144 In-vitro and in-vivo antibacterial effect of Croton lobatus Linnaeus L. 54(2) 2019

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative method was used to estimate total phenol contents of the alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family parasitic infection, treatment for menstrual cycle disorders, Examinations of Antimicrobial activity of the C. lobatus Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West Preparation of crude extract Conclusion Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water of Science and Technology 5(1): 1936- 1940. Phytochemical Studies Aké AL (1975), Contribution aux études ethnobotaniques Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its ent-Beyer-15-en-18-oic acid, Pharmaceutical This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into uses, Chemistry and Pharmacology of Croton species dorsal region was surgically prepared for aseptic surgery and glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 Kilani, Hassan, Fadason, Obalowu, Aliyu and Badmus-Kilani 145

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of method was used to estimate total phenol contents of the feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family parasitic infection, treatment for menstrual cycle disorders, Examinations of Antimicrobial activity of the C. lobatus Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West Preparation of crude extract Conclusion Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water of Science and Technology 5(1): 1936- 1940. Phytochemical Studies Aké AL (1975), Contribution aux études ethnobotaniques Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into ent-Beyer-15-en-18-oic acid, Pharmaceutical dorsal region was surgically prepared for aseptic surgery and uses, Chemistry and Pharmacology of Croton species glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021 146 In-vitro and in-vivo antibacterial effect of Croton lobatus Linnaeus L. 54(2) 2019

In Africa, C. lobatus is commonly used by Indigenous people Methods and materials Quantitative analysis of the phytochemicals tannins as contained in other species of Croton (Attioua, count was recorded with physiological normal saline treated reveals that C. lobatus is very active against organisms that Fakhim et al., 2015), triterpenoids and flavonoids (Akpalo et Amaral ACF and Barnes RA (1998), A Dadson YP, Tikkanen-Kaukanen C and Miettola J (2012), Michalik L and Wahli W (2007), Peroxisome (Euphorbiaceae), Journal of Brazilian Chemical Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey to treat different diseases such as malaria, diarrhea and 2005; Burkill, 1985; Chabert et al., 2006; Farnsworth, 1969; wounds and C. lobatus water extract (Fig. 4). grow best in nutrient agar like Bacillus spp, Streptococcus al., 2015; Thakur et al., 2011) crotonic acids (Goldstein et tetrahydroprotoberberine alkaloid from Croton Antibacterial properties of Croton species. Institute of proliferator-activated receptors (PPARs) in skin health, Society 18(1): 11-33. DOI:org/10.1590/S0103- J, Gbenou J, Moudachirou M, Brun R, Anton R and problems due to pregnancy (Aké, 1975). The crude extracts Sample Collection Flavonoids, alkaloid saponin and tannin were determined as Willaman and Li, 1970). Croton lobatus L. contained 5.22 % spp, E. coli, Pseudomonas and Staphylococcus spp. Hence, it al.,, 2003; Michalik and Wahli, 2007) and saponin (Thakur et hemiargyreus, Phytochemistry 47: 1445. Public Health and Clinical Nutrition, Faculty of Health repair and disease, Biochim Biophys Acta (BBA) 50532007 000100002 Sanni A (2004), Evaluation of ethnobotanically from the aerial and roots of C. lobatus have anti-plasmodia describes by Bohm and Kocipai-Abyazan (1994), Harborne alkaloid that was classified into five different types by The in vitro antimicrobial assay of C. lobatus plant extracts is active against microbial wound infections thereby al., 2011) that is present in the leaves. This finding is DOI:org/10.1016/S0031-9422(97)00764-4 Sciences, University of Eastern Finland. Molecular and Cell Biology of Lipids 1771(8): selected Benin medicinal plants for their in vitro Croton lobatus L. was collected from Obokun Local activity (Weniger et al. 2004). Heated leaves are rubbed on to (1973), Obadoni and Ochuko (2001) and Van-Burden and Barthelemy et al. (2012). The study also revealed that C. showed a dose depended activity against the tested preventing delayed wound healing. The antimicrobial supported by several other scientists who reported the 991–998. DOI:org/10.1016/j.bbalip.2007.02.004 Shinwari ZKI, Khan S and Hussain NA (2009), Assessment antiplasmodial activity, Journal of Government Area of Osun state Nigeria and was confirmed in Attioua B (2005), Contribution à l’étude phytocimique des areas of costal and rheumatic pain, and a leaf-decoction by Robinson (1981) respectively. While the spectrophotometric lobatus contains steroids and triterpenes, flavonoids, microorganisms while the in vivo antimicrobial studies property can be attributed to tannins (Christian et al., 2016; antimicrobial activities of other species of croton (Abo et al., Evans WC and Trease GE (1996), Trease and Evan of antibacterial activity of three plants used in Pakistan Ethnopharmacology 90: 279-284. DOI:org/10.1016/ the Botanical Garden, Biological Science department of method was used to estimate total phenol contents of the feuilles et tiges de Croton lobatus Linn. th Milanowski DJ, Winter REK, Elvin-Lewis MPF and Lewis to cure respiratory diseases, African Journal of mouth, or a bark-decoction by enema is given as a purgative alkaloids, and saponin similar to other species of croton. 1999; Dadson et al., 2012; McChesney et al., 1991; Peres et Pharmacognosy, 14 Ed. WB, Sounders Co. Ltd, j.jep.2003.10.002 Ahmadu Bello University, Zaria, with voucher specimen plant. (Euphorbiaceae), Ph.D. Dissertation, University of WH (2002), Molecular Cloning and Characterization Biotechnology 8(24): 7082-7086. (Burkill, 1985). The plant is used generally as healing leaf Similar studies have revealed the presence of triterpenes al., 1997; Salatino et al., 2007). London, pp 542-578. medicine, pain-killer, laxative, cutaneous and subcutaneous number of 913. (Barbosa et al., 2003), steroids, flavonoids (Cai et al., 1991; Strasbourg, p 135. of CYP80G2, Journal Natural Product 65: 814. Willaman and Li (1970), C. lobatus Linn. family parasitic infection, treatment for menstrual cycle disorders, Examinations of Antimicrobial activity of the C. lobatus Salatino et al., 2007) alkaloids (Aboagye et al., 2000; Fakhim SA, Babaei H, Nia AK and Ashrafi J (2015), Wound Sofowara A (1993), Medicinal plants and Traditional Euphorbiaceae In: The Useful Plants of West Preparation of crude extract Conclusion Barbosa PR, Fascio M, Martins DS, Guedes ML and Roque F antidotes (venomous stings and bites), treatment for Amaral and Barnes, 1998; Milanowski et al., 2002; Healing Effect of Topical Grape Seed Extract (Vitis Nath R, Roy S, De B and Choudhury MD (2013), Anticancer medicine in Africa, Spectrum Books Ltd, Ibadan, Tropical Africa (2), Burkill HM, Royal Botanic Microbial isolate used were Staphylococcus aureus (ATCC (2003), Triterpenes of Croton betulaster paralysis, epilepsy, convulsions, spasm and topically to treat Yibralign, 2007) triterpenoids, either pentacyclic or steroidal This study has revealed that C. lobatus contains secondary Vinifera) on rat palatal mucosa, International Journal and Antioxidant activity of Croton: A Review, Nigeria, p 289. Gardens, Kew (K), Leave of C. lobatus was harvested and air dried at normal 33591); Streptococcus Spp (By Dr Raji bacteria zoonosis (Euphorbiaceae), Biochemical Systematics and International Journal of Pharmacy and ulcers, sores, and headache (Burkill, 1985). Odukoya et al. (Nath et al. 2013) and saponin (PROTA, 2011). metabolites such as carbohydrates, glycoside, saponins, of Current Research and Academic Review 3(6): temperature. The dried powder of the plant was extracted laboratory, A.B.U); Pseudomonas aeruginosa (ATCC 9028); Ecology 313: 07–308. Pharmaceutical Sciences 5(2): 63-70. Thakur R, Jain N, Pathak R and Sandhu SS (2011), Practices Yibralign Z (2007), Phytochemical Investigation on the stem (2012) reported that C. lobatus is usually boiled with water steroids, triterpenes, flavonoids and tannins, which are 477–489. with distilled for 24 h. The extract was filtered through Proteus vulgaris (isolate identified from bacteria zoonosis in Wound Healing Studies of Plants, Evidence-Based bark of Croton macrostachyuss (Bisana), Master’s and the decoction is taken to treat wounds that refuse to heal Antibacterial activities responsible for its antimicrobial property. Hence, this Barthelemy KA, Ramiarantsoa H, Jean BB, Vincent AA, Whatmann filter paper No 1 and it was thereafter, centrifuged laboratory VPH, A.B.U); Escherichia coli (ATCC 43895); Farnsworth (1969), Croton lobatus Linn. Linn. family Obadoni BO and Ochuko PO (2001), Phytochemical studies Complementary and Alternative Medicine, 438056. Thesis. Addis Ababa University, Addis Ababa. while Bouquet and Debray, (1974) reported the whole plant explains its usage by the local medicine practitioner in Félix ZT and Léon AD (2012), Isolation and at 5000G for 15 min. The supernatant were further filtered Salmonella Spp (ATCC 4932). Croton lobatus L. showed a dose dependent activity against Euphorbiaceae In: The Useful Plants of West and comparative efficacy of the crude extract of some DOI:org/10.1155/2011/438056 infusion to be used as a topical bath to treat skin diseases. through Millipore filter before storage at 4° C as described by Nigeria and various part of the world as alternative identification of alkaloids from C. lobatus, micro organisms. Croton lobatus paste and C. lobatus 0.5 Tropical Africa (2), Burkill HM, Royal Botanic homeostatic plants in Edo and Delta states of Nigeria, (Prashanthi et al., 2012). therapy in the management of diseases whose symptoms International Journal of Pharmaceutical Sciences In vitro antimicrobial assay retarded the growth of Staphylococcus aureus; Gardens, Kew, UK. Global journal of Pure and Applied Sciences 8: Van-Burden TP and Robinson T (1981), The biochemistry of Croton lobatus L. has recently been reported by traditional 13(2): 001 nd Streptococcus Spp; Pseudomonas aeruginosa; Proteus involve microbial infections. Further research is required 203-208. DOI:org/10.4314/gjpas.v8i2.16033 alkaloids. 2 Ed. Springer, Heidelberg, Newyork. medicine practitioner in Nigeria as having a good therapeutic Preliminary phytochemical test A method by Irobi et al. (1994) and Shinwari et al. (2009) vulgaris; E. coli and Salmonella Spp at the zone of on this plant. Gaikwad SP, Gore RD and Garad KU (2012), Astraea lobata index on chronic wounds by applying its fresh leaves or dried was adopted. The prepared C. lobatus water extract was Block S, Baccelli C, Tinant B, Meervelt LC, Rozenberg R, inhibition ≤ 21mm. The C. lobatus 0.05 inhibited the growth (Euphorbiaceae), a new record for India, Rheedea Odukoya OA, Sofidiya MO, Samuel AT, Ajose I, Onalo M powder on wound surface while the leaf-decoction is used to Screening for the phyto-constituents such as saponins, reconstituted with sterile distilled water similar to what was Jiwan JLH, Llabrès G, Pauw-Gillet MC and of Proteus vulgaris at 34mm while C. lobatus 0.005 Acknowledgment 22(2): 131-132. and Shuaib B (2012), Documentation of Wound aid development of fetus. Hence, it is necessary to screen for alkaloids, flavonoids, steroids, tannins, cardiac glycosides, done by Ongsakul et al. (2009) to obtain a concentration of Quetin-Leclercq J (2004), Effects of Croton inhibited the growth of Staphylococcus aureus at 18mm Healing Plants in Lagos-Nigeria: Inhibition of Lipid the phytoconstituents of C. lobatus and evaluate its glycosides, and proteins was carried out as described by 0.5g/mL. Lower concentrations of 0.05 g/mL and 0.00 5g/mL indelible marker was used to make a sterile square shape (0.5 Authors acknowledge the effort of the department of the rhamnifolioides Essential Oil on Aedes, (Fig. 2). CFU counted on nutrient agar reduced drastically Global Biodiversity Information Facility, GBIF (2016), Peroxidation as In-vivoPrognostic Biomarkers of antimicrobial activities against common microorganisms that (Evans and Trease, 1996; Harborne, 1973; Sofowara, 1993). of water were then prepared from the concentration of by 0.5 centimeter) in four different places (Fig. 1). Veterinary Public Health and Preventive Medicine; and Phytochemistry 65: 1165. after initiation of treatment (day 2) in all the treated wounds Backbone Taxonomy. Available at https://www. Activity, Annals of Biological Research 3(4): infect wounds. 0.5g/mL (Table I). Broth cultures with turbidity equivalent of Full-thickness skin incision was carried out to create the Department of Pharmacognosis, Faculty of Pharmacy, (A, B and D) except with physiological normal saline Bohm BA and Kocipai-Abyazan R (1994), Flavonoids and gbif.org. 20: 17. 1683-1789 0.5 McFarland standards of known bacteria strains as listed wounds. The wounds were left for 48 hrs to mimic the clinical treated wounds (Fig. 3). On the MacConkey agar, CFU Ahmadu Bello University Zaria. Dr Raji (bacteria zoonosis condensed tannins from the leaves of Vaccinum earlier were prepared. Using a sterile cotton swab stick, the condition, after which they were accessed and samples were Ongsakul M, Jundarat A and Rojanaworant C (2009), counted significantly reduced with days on cicatrin treated laboratory, A.B.U) is also acknowledged for kind donation of raticulation and Vaccinum calcyimium, Pacific Science Goldstein JT, Dobrzyn A, Clagett-Dame M, Pike JW and Mueller Hinton Agar (MHA) Plates were inoculated. Sterile collected for culture, identification and CFU. Respective Antibacterial Effect of Crude Alcoholic and Aqueous wounds, slightly reduces with days on the wounds treated Streptococcus sp. of bacteria used. 48: 458-463. DeLuca HF (2003), Isolation and characterization of cork borer was used to create wells of 5 mm diameter in the treatment using C. lobatus (paste), C. lobatus (water extract), Extracts of six medicinal plants against Staphylococcus with C. lobatus paste while persistent high number of CFU unsaturated fatty acids as natural ligands for the medium and 100 μl of C. lobatus 0.5g/mL, 0.05 g/mL and Physiological saline solution (PSS), Cicatrin powder was References Bouquet A and Debray M (1974), Medicinal plants of the retinoid-X receptor, Archives of Biochemistry and aureus and Escherichia coli, Journal of Health carried out after 48 hrs of wound creation (Table I) and 0.005 g/mL extracts were filled into each of the wells. The Ivory Coast, Travaux Ed Qocumentes de I’orstom Biophysics 420(1): 185-193. DOI:org/10.1016/j.abb. Research 23(30): 153-156. samples were collected at day one, three, five and seven after plates were incubated at 37°C for 24 hrs. The diameters of the Abo KA, Ogunleye VO and Ohidi JS (1999), Antimicrobial 32: 232. 2003.09.034 growth inhibition formed in mm were recorded. The zone of initiation of treatments for CFU counts in nutrient agar and potential of Spondias mombin, Croton zambesicus and Peres MT, Monache DF, Cruz AB, Pizzolatti MG and Yunes inhibition was compared with standard antibiotics (Table I). MacConkey agar. Zygotritonia crocea, Phytotherapy Research 13(6): Burkill HM (1985), Croton lobatus Linn, family Harborne JB (1973), Phytochemcial methods, London, RA (1997), Chemical composition and antimicrobial 494-7. Euphorbiaceae In: The Useful Plants of West Tropical Chapman and Hall Ltd., pp 49-188. activity of Croton urucurana Baillon (Euphorbiaceae), In vivo antimicrobial assay Data analysis Africa (2), Royal Botanic Gardens, Kew (K), Journal of Ethnopharmacology 56(3): 223-6. Aboagye FA, Sam GH, Massiot GL and Julocrotine CA Irobi ON, Moo-Young M, Anderson WA and Daramola SO DOI:org/10.1016/S0378-8741(97)00039-1 The results were presented inform of tables and charts, with Cai Y, Evans F, Roberts M, Phillipson JD, Zenk M and Gleba A male albino rat of 22 weeks old, weighing about 200g was (2000), Glutarimide alkaloid from Croton (1994), Antimicrobial Activity of the Bark of Bridelia bars representing standard error of means. Y (1991), Effects of Sangre de Drago from Croton selected from an inbred colony obtained from Jos, Plateu membranaceus Fitoterapia 71: 461-462. fermginea (Euphorbiaceae), Interational Journal of Prashanthi R, Mohan N and Siva GV (2012), Wound healing lechleri Muell, Phytochemistry 30: 2033. State. It was housed in a steal cage under normal temperature, DOI:org/10.1016/S0367-326X (00)00141-6 Pharmacology 34: 87-90. DOI:org/10.1016/ property of aqueous extract of seed and outer layer of humidity and light. The rat was fed with commercial rat feed Results and discussion DOI:org/10.1016/S0378- 8741(97)00087-1 Momordica charantia L. on albino rats, Indian Journal 0378-8741 (94)90041-8 (prepared from vital feed grower mash) and adequate water of Science and Technology 5(1): 1936- 1940. Phytochemical Studies Aké AL (1975), Contribution aux études ethnobotaniques Chabert P, Attioua B, Weniger B and Brouillard R (2006), was provided. Ethical permission was granted by the Ahmadu et floristiques au Togo, Collection “Médecine Malpighiales of North America Update, database (2011), Croton lobatus Linn., an African Medicinal plant: PROTA (Plant Resources of Tropical Africa) (2011), Croton Bello University Committee on Animal Use and Care. The The qualitative studies on the leaves of C. lobatus water traditionnelle et pharmacopée, Agence de Retrieved from http://www.itis.gov/servlet/ Spectroscopic and chemical elucidation of its many macrostachyus Hochst. ex Delile. Available at permission number was ABUCAUC/2016/027. The animal extracts revealed that it contains carbohydrates, glycoside, Coopération Culturelle et Technique 153. SingleRpt/SingleRpt? On August 10, 2016 at 22:30 constituents, Bio Factors 27: 69-78. http://www.prota4u.org/protav8.asp?h=M4&t=Croton was allowed to acclimatize for two weeks before the saponins, steroids, triterpenes, flavonoids, alkaloids and GMT &p=Croton+macrostachyus#Synonyms. Accesed on experiment. It was anesthetized by intra muscular injection of tannins (Table II). The percentage composition of the Akpalo E, Tete-Benissan A, Awaga K and Akpagana K Christian A, Bekoe EO, Boakye YD, Dapaah SO, Appiah T (R) 07.012.2015 at 22:00GMT. Rompun (xylazine hydrochloride) at the dosage of 5 mg/kg alkaloids, flavonoids, saponins, phenols and tannins in the (2015), Review of twelve West Africa Medicinal and Bekoe SO (2016), Medicinal Plants and Natural McChesney JD, Clark AM and Silveira ER (1991), and ketamine hydrochloride at the dosage of 40 mg/kg. The leaves of C. lobatus water extract is presented in the Table II. Plants: Active phytochemical combinations in direct Products with Demonstrated Wound Healing Antimicrobial diterpenes of Croton sonderianus. II. (R) Salatino A, Salatino MLF and Negri G (2007), Traditional whole body of the animal was cleaned with diluted Purit . Its This study revealed that C. lobatus contains carbohydrates, biochemically wound healing process, Journal of Properties In: Wound Healing - New insights into ent-Beyer-15-en-18-oic acid, Pharmaceutical dorsal region was surgically prepared for aseptic surgery and uses, Chemistry and Pharmacology of Croton species glycoside, saponins, steroids, triterpenes, flavonoids and Medicinal Plant Research 9(34): 908-917. Ancient Challenges, INTECH 483-531. Research 8(10): 1243-7. DOI: 10.1021/np50078a021