African J. Biol. Sci., 15 (1): 101-115 (2019) ISSN 1687-4870 e- ISSN 2314-5501 (online) www.aasd.byethost13.com e.mail: [email protected]

Control of the glassy clover snails cartusiana using Zingiber officinale extract as an ecofriendly molluscicide

Mahmoud Abd El-Atti1*, Elsheakh A. A.2 , Khalil A.M.1 and Wesam S. Elgohary2 1- Zoology Department, Faculty of Science, Zagazig University, Zagazig. Egypt 2- Plant Protection Research Institute, Agr. Res. Center. Doki, Giza, Egypt *Corresponding author email: [email protected]

ABSTRACT High population densities of Monacha cartusiana were observed to cause perdition of various economic crops at Sharkia Governorate. The present study was carried out control this harmful pest using ethanolic Ginger extract as a natural and environmentally safe molluscicide.Toxicological tests revealed that low concentration (20%) of ethanolic Ginger extract caused 10 % mortalityof cloversnails after one day of exposure and 66.7% mortalityafter28 days. The highest mortality percentage (90 %) was recorded after 28 days of treatment with40% Ginger extract. Biochemical investigations showed thatLC25 (7.04%) Ginger extract elevated ALT, AST, α & β esterases andphenoloxidase levels of treated snails compared to control.Histological inspections of the digestive gland and ovotestis of snails exposed to LC25Ginger extract revealed that the digestive tubules showedvarious tubularanomalies, inflammatory hemocytes infiltration, excessive luminal secretionsand appearanceof necrotic areas. Acini of ovotestis weredeformedand comprised misshapen eggs and distorted sperms.TEM investigations showed cellular malformations in the digestive gland of treated snails including; severely disrupted microvilli, ruptured cell membranes, fractured RER and mitochondrial pyknosis.

Key words: Ecofreindly- Antioxidant Enzymes- Ginger extract-Histopathology- Monacha cartusiana-TEM.

INTRODUCTION 2019). High population densities were Land snails are considered as serious discoveredinfesting various economic crops agricultural pests Egypt. They are at Sharkia Governorate, Egypt (Lokma, herbivorous that devour mellow 2013). M. cartusiana reported to act also as parts of vegetables, fruit trees and an intermediate host for sheep lungworm ornamental plants (Godan, 1983) and ruin and lancet liver fluke (Godan, 1983). mushy leaves, fruits, seeds and tubers (El- Chemical control applications still the most Okda, 1984). Undesirable smell and savor commonly used for controlling land caused by mucous secretions left on plants snailsalthough; they extensively prejudice during their movements, averts human and non-target organisms including man (Gabr other animals from feedingon these et al., 2006). The high costs and resistance contaminated plants (El-Deeb et al., 1999). exerted against synthetic pesticides The glassy clover snails (Monach encourage the usage of naturally derived cartusiana) were recorded extensively at pest control strategies (Massoud and Habib vast geographic areas at the Mediterranean 2003). Recently, several countries enhanced regions and Southeastern Europe the use of plant extracts in pest control due (Pieńkowska, 2018: Pieńkowska et al., to their low mammalian toxicity, low costs

102 Mahmoud Abd El-Atti et al. and fast biodegradability (Singh et al., 3. Experimental design 2000). Ginger extracts possess anti- Clover snails were divided randomly into inflammatory, analgesic, antioxidants and three main groups; the first group used for antimicrobial activities (Sharma et al., 2013; toxicological studies was subdivided into five Nikoli et al., 2014; Amri and Touil- sub groups and treated with different Boukoffa, 2016). Few studies have been concentrations of Ginger extract for 28 days. carried out on the molluscicidal activities of Four replicates were applied for each Ginger (Ahmad et al., 2013; and Edwin and subgroup.The second groupfed with baits Jacob 2017). Physiological and containing LC25 of ginger extract for 14 days histopathological investigations give andused for biochemical, histopathological and excellent indication about the hazards occurs ultrastructural investigations.The third group left within the organism because of exposure to as control group. toxicants. The present study aimed to control the glassy clover snail Monacha 4. Biochemical measurements: cartusiana using Ginger extract as a natural Digestive glands were dissected out from and alternative solution instead of using both control and treated snailsthen homogenized harmful chemical molluscicides. in distilled water using a Teflon homogenizer. The homogenates were centrifuged at 8000 MATERIALS AND METHODS r.p.m. for 15 minutes at 5ºC inrefrigeratedcentrifuge. Deposits were 1. Snails collection and acclimation discarded and supernatants were kept in Adult snails of M. cartusiana were a deep freezer untiluse. Activities of AST collected from clover fields at Malames and ALT enzymes were determined according to village, Meniet El-kamh district, Sharkia the procedure of Reitman and Frankle (1957). Governorate during spring 2018. Snailswere Alpha and Beta esterases assayed according to transferred to the laboratory and kept in Van Asperen, (1962). Phenoloxidase glass cages (50 x 30 x 30 cm) contained activitymeasured using the method of Ishaaya moist soil and fed daily with fresh Lettuce (1971).All biochemical measurements were for two weeks. carried out in Plant Protection Research Institute, Agriculture Research Centre, Giza, Egypt. 2. Ginger extracts preparation Rhizomes of Ginger were purchased 5. Histopathological studies from the InternationalCompany (Cairo- For light microscopy, digestive glands of Egypt). Rhizomeswere ground intoa fine both control as well as treated with LC25 Ginger powder using a pestle and mortar. The extract for 14 days were dissected out and fixed powder (30 g) was soaked in ethanol (600 in Bouin's fluid. Specimens dehydrated in a ml) in a Sechelt apparatus for twodays. graded series ofethanol, cleared in Xylene for 20 Ethanol was evaporated under minutes, impregnated in paraffin wax (three reducedpressure until brown extract was changes) at 60oC for 2 hours and embedded in obtained (Bakry et al., 2013). Poisonous paraffin wax. Sections (4-5 µm thick) were cut baits were formulated by mixing ethanolic by microtome, mounted on glass slides Ginger extract with five grams of sugarcane andstained with hematoxylin and eosin. syrup, completed with wheat bran to 100 grams and moisten with small amounts of 6. Ultrastructural studies water. Dissected digestive glands of both control and Ginger treated snails were fixed in 2.5 % glutaraldehyde in 0.05 M cocodylate

103 Control of the glassy clover snails Monacha cartusiana using Zingiber officinale extract as an ecofriendly molluscicide buffer containing 0.15 sucrose at pH 7 for RESULTS two hours. Specimens were fixed in 1% 1. Toxicity tests Osmium tetroxide at 4oC for an hour, Figure (1) showed that snail dehydrated in an ascending series of ethanol mortalities increased with increasing of both and embedded in Araldite f Epon. Blocks Ginger extract concentrations and were cut with diamond ultramicrotome exposuretime. At the first day of exposure, knifes. Semithin sections stained with 1% all treatments caused low mortalities. Toluidine blue. Ultra-thin sections stained Ethanolic extract of Ginger (20%) causes with aqueous Urinyl acetate and Lead citrate 10, 50, 56.6, 66 and 66.7% mortalitiesafter and examined under JEOL 100 CX TEM at 1, 7, 14, 21 and 28 days of treatment the Electron Microscopy Unit, Faculty of respectively.Exposure to 30 % Ginger Agriculture, El-Mansoura University, Egypt. extract caused 6.6, 36.7, 70, 76.6 and 83.3% for the same days of 7. Statistical analysis: exposurerespectively.Finally, 40% Ginger Values of toxicological and extract resulted in 10, 46.6, 76.6, 83.3 and biochemical studies were expressed as a 90% respectively, at the same time intervals. mean ± SE. The obtained data were analyzed The highest percentage of mortalities (90%) statistically for the significance of differences occurred after treatment with40% Ginger using student’s t-test (Goldstein, 1964). extract for 28 days (Fig. 1).

Fig.(1): Mortality percentages of M. cartusiana exposed to different concentrations of Ginger extract for 28 days.

2. Biochemical measurements their levels slightly decreased with 2.1. Activities of ALT and AST increasing time of exposure, although levels Levels of AST were significantly were much higher than control (Fig. 2). elevated (P <0.05) after exposure to LC25 (7.04%) of Z. officinale extract and the 2.2. Activities of α and β-esterases highest level was recorded after two weeks LC25 of Z. officinale extract caused a of exposure compared to control. Activities highly significant increase (P <0.05) of α of ALT were significantly increased(P < and β-esterases activities after the first day 0.05 ) after the first day of treatment, while of exposure. Values of both enzymes were

104 Mahmoud Abd El-Atti et al. decreased insignificantly (P>0.05) after the following the first day of treatment. 7th and 14th day of treatment (Fig. 2). Phenoloxidase levels decreased insignificantly (P>0.05) after one week of 2.3. Activity of Phenoloxidase treatment. The lowest level of phenoloxidase Highly significant increase in the was achieved after two weeks of exposure to activity of Phenoloxidase was detected Ginger extract (Fig. 2).

Fig. (2): Enzymatic activities of M. cartusiana treated with LC25 Ginger extract for 14 days.

3. Microscopical Examination Excretory cells: Rounded cells incorporate 3.1. Histological studies on control large dark brown excretory granule and digestive and hermaphrodite glands small basally locatednuclei. The digestive gland of control M. Calcium cells: Occur singly or in groups, cartusianacomprises mainly manynumerous pyramidal-shape, stacked with rounded blind digestive tubules separated by calcium spherules (appeared as light bodies) intertubular connective tissue containing and have centrally located globular nuclei. hemolymphatic vessels and disparate On the other hand, the hermaphrodite hemocytes. Each tubule encircled with gland (ovotestis) is embedded in the circular muscle layer (Plate I, Fig. a). Three digestive gland and consists of many different cell types constitute the epithelial ovoidacinieach is bounded by interacinar lining of digestive tubules: digestive, connective tissue containing interstitial cells. excretory and calcium cells (Figs. b- d). Each acinus isborderedby germinal Digestive cells: Columnar cells, possessed epithelial layerand many Sertoli cells in many vacuolesaccommodating yellowish between. Their lumens embrace brown granules and basally located rounded developmental stages of spermatogenesis or oval nuclei. and oogenesis (Figs. e &f).

105 Control of the glassy clover snails Monacha cartusiana using Zingiber officinale extract as an ecofriendly molluscicide

Plate I. Photomicrographs showing sections of the digestive and hermaphrodite glands of control M. cartusiana. (a):T.S. of untreated digestive gland showing the normal histological architectureof digestive tubules (X100). (b): Semi thin section showing different types of cells forming digestive tubule and calcium spherules of calcium cells (X400). (c & d): Sections of the digestive tubules showing calcium cell at the corners, yellowish brown granules of digestive cells and large excretory dark granule of excretory cells (X400). (e & F): Cross sections showing ovotestis acini of control M. cartusiana containing different developmental stages of oocytes and sperms (X400). ACI: Acini of the ovotestis; CC: Calcium cell; DC: Digestive cell; DT: Digestive tubules; EC: Excretory cell; GE: Germinal epithelium; HE: Hemocytes; ICT: Intertubular connective tissue; IS: Interacinar space; ISC: Interstitial cell; L: Lumen; ML: Muscle layer; PO: Primary oocyte; SO: Secondary oocyte; Sd: Spermatids; SP: Sperms; SR: Sertoli cell; YBG: Yellowish brown granules.

106 Mahmoud Abd El-Atti et al.

3.2. Ultrastructural studies of control electron- lighted calcium spherules, electron digestive gland dense granules and oval mitochondria which Transmission electron microscopic are more concentrated at the apex (Fig. d). studies revealed that the hepatic ducts of Excretory cells: Rounded cells contain M.cartusiana digestive glandarelined with a massive microvilli in their free border. The ciliated columnar epithelium resting on a nucleus is rounded, basally located and thin basement membrane andsupported by surrounded bydense layer of rough connective tissue layer and circular muscle endoplasmic reticulum and numerous oval bundles. These cells have oval nucleiwith mitochondria. The cytoplasm filled with prominent nucleolus and surrounded with dark osmiophilic excretory granules. A thin layer of rough endoplasmic reticulum. smaller excretory cell (regenerative) was Their cytoplasm is homogenous and noted beside the mature one that has a large contains many pinocytotic vesicles, globular nucleus containing an eccentric whichwere noted also in the underlying nucleolus. Its cytoplasm is condensed, connective tissue layer. Mitochondria are containing few mitochondria and devoid of oval and more concentrated at the apex of excretory granules (Figs. e& f). these cells (plate II, Figs. a&b).Electron micrographs confirmed the presence of three 3.3.3. Treated digestive gland main cell types constituting the epithelial Light microscopic examinations of lining of the tubules which are differentiated the digestive gland treated with into; digestive, excretory and calcium cells. LC25ofGinger extract revealed various Digestive cells: Their outer borders tubular deteriorations, inflammatory provided with well-developed microvilli hemocytic infiltrations, tissue exudates and projecting towards the lumenand excessive luminal secretions. The epithelial havebasally ovalnuclei. Mitochondria and lining undergoes vacuolization, extensive rough endoplasmic reticulum are scattered cell lysis and necrotic areas. The basement throughout cytoplasm. The cytoplasm membranes and muscular layers surrounding contains alsomany electron- dense tubules were lacerated. The digestive cells lysosomalgranules showing different states have accumulations of large numbers of of activities. Adjacent digestive cells are darkly stained granules and pyknotic nuclei. interconnected by inter inter-cellular Excretory cells showed decreased amount of junctions (Fig.c). dark brown granules. Calcium cells were Calcium cells: Pyramidal cells have disrupted and have few number of calcium numerous apical microvilliand rounded spherules (Plate III, Figs. a-d). nuclei. Their cytoplasm possesses many

107 Control of the glassy clover snails Monacha cartusiana using Zingiber officinale extract as an ecofriendly molluscicide

Plate II. Electron micrographs showing normal cellular structures of the digestive gland of M. cartusiana.(a): Ciliated columnar cells lining the hepatic duct (X13600). (b): Connective tissue layer and circular muscle bumbles surrounding digestive tubules (X17000). (c): Cellular lining of digestive tubule showing group of digestive cells (X 6800). (d): Calcium cells containing characteristic calcium spherules (X6800). (e): The basal region of excretory cell and secretory granules (X17000). (f): Small regenerative excretory cell (X17000). BM: Basement membrane; C: Cytoplasm; CC: Calcium cells; CH: Chromatin material ; CS: Calcium spherules; CI: Ciliated columnar epithelium; CMB: Circular muscle bundles; CT: connective tissue; DC: Digestive cell ; DV: Digestive vesicles; EC: Excretory cells; EG: Excretory granules; EEC: Regenerative (Embryonic) excretory cell; HD: Hepatic duct; ICJ: Inter cellular junctions; ICT: Intertubular connective tissue; L: Lumen; LV: Lysosomal vesicles; M: Mitochondria; MV: Microvilli; N: Nucleus; NU: Nucleolus; PV: Pinocytotic vesicles; RER: Rough endoplasmic reticulum.

108 Mahmoud Abd El-Atti et al.

Plate III. (a- c): Photomicrograph showing cross sections through the digestive gland of M. cartusiana treated with LC25 of ethanolic Ginger extract for 2 weeks(X 400). (d): T. S. of treated ovotestis showing deformed oocyte and sperms(X 400). CL: Cell lysis; CS: Calcium spherules; DBM: Degenerated basement membrane; DCC: Deformed calcium cells; DEC: Deformed excretory cells; DDT: Deformed digestive tubules; DG: Dark granules; DSP: Deformed spermatozoa; EC: Excretory cell; ES: Excessive secretions; ITF: Intertubular filtrate; HA: Hermaphroditic acinus; HI: Hemocytic infiltration; N: Nucleus; NA: Necrotic area; PN: Pyknotic nuclei; S: Secretions; YG: Yellowish brown granules.

3.4. Ultrastructural abnormalities cellular debris. RER of excretory and Electron micrographs of the calcium cells were ruptured in the form of digestive gland of M. cartusiana treated disorganized tubules. .Mitochondria showed with LC25 Ginger illustrated many condensation of their matrix (mitochondrial ultrastructural abnormalities as severely pyknosis) andsprinkled cristaeand the nuclei ruptured microvilli of digestive and calcium became pyknotic. Intercellular junctions of cells.Excessive cloudy luminal secretions adjacent cells were deformed (Plate IV, were noted. Cell membranes of various cells Figs. a- d). were lacerated. The cytoplasm was highly vacuolated inclosing denselyosmiophilic granules and lysosomal vesiclescomprising

African J. Biol. Sci., 15 (1): 101-115 (2019) ISSN 1687-4870 e- ISSN 2314-5501 (online) www.aasd.byethost13.com e.mail: [email protected]

Plate IV. (a) Electron micrographs showing ultrastructural alterations in the digestive gland of M. cartusiana treated with LC25 Ginger ethanolic extract. (a):Apical portion of digestive cellshowing disrupted microvilli(X 10200). (b): Basal region of excretory cellshowing nuclear pyknosis and vacuolization (X 13600). (c): Digestive cell with distorted brush boarder and pyknotic mitochondria (X = 10200). (d): Calcium cell with lytic cytoplasm, ruptured RER and microvilli (X = 17000). CD" Cellular debris; Ch: Chromatin materials; DMV: Disrupted microvilli; DOG: Darkly osmiophilic granules; FOG: Faint osmiophilic granules; LC: Lytic cytoplasm; LS: Luminal secretions; LV: Lysosomal vesicles; M: Mitochondria; PN: Pyknotic nucleus; RCM: Ruptured cell membrane; RCJ: Ruptured intercellular junctions; RER: Deformed rough endoplasmic reticulum; V: Vacuoles.

DISCUSSION 2017). The present study showed that low Several toxicological studies have concentrations of ethanolic Ginger extract been carried out to evaluate the caused low mortality percentages after the molluscicidal activities of chemical first day of exposure while the highest pesticides, plant extracts and biocides mortalities of M. cartusiana snails occurred against harmful land snails (Asran, 2001; at high concentrations of Ginger at the third Gabr et al., 2006; El-Sherbini et al., 2009; and fourth weeks.The presence of alkaloids, Abdel-Haleem, 2013; EL-Sayed et al., 2013 flavonoids, tannins, saponins, terpenoids and Mwonga et al., 2015; Prabhakaran et al., phenolic derivativescaused the molluscicidal

110 Mahmoud Abd El-Atti et al. potency of Ginger (Sharma et al., 2016). enzymeand plays an important role in the This finding was in harmony with Singh et immune system of most invertebrates (Smith al. (2010) who revealed that ceniole, citral, 6 and Soderhall, 1991; Soderhall and - gingerol and oleoresin extracted from Z. Cerenius, 1998). The activity of Phenol officinale rhizomes had potent molluscicidal oxidases were stimulated after the entrance properties and these active ingredients of exotoxins inside organisms (Lee et al., resulted in an inhibition in the reproductive 1998). Ginger ethanolic extract caused capacity of Lymnaea acuminate snails. significant increase in the PO activity for a Bakry et al. (2013) showed a rapid decline week then decreased at the second week. in survival rate and egg production of This result may attributed to the Biomphalaria alexandrina snails disturbances occurred in some defensive exposed to Ginger. The obtained results mechanisms of the snail as a result to the showed that Ginger extract caused high stress induced by Ginger extract. These disturbancesin the enzymatic activities of results are in conformity with Farrag et al. treated clover snails. Highly significant (2015) who stated that Phenoloxidase levels elevationsof AST and ALT levelsrecorded were significantly increased in treated 5th after the first day of exposure may be nymphal instar haemolymph of desert locust attributed to the activation of defensive after two dayswhile at the fourth days of mechanisms inside the snail body to treatment with Metarhizium anisopliae, their overcome the effect of toxicants. The levels were decreased significantly gradual decrease happened in both enzymes compared to control. later may be duo to cell injuries in digestive The digestive gland (liver) of gland of treated snail and toxical hepatitis molluscs is involved in digestion, absorption (Farkas et al., 2004). Results demonstrated beside storage of lipids, glycogen and that Ginger extract caused a significant minerals (Beeby and Richmond, 1988), and increase in α- esterase while it caused initial plays a major role in detoxication (Henry et increase in β-esterase after one day of al., 1991).The digestive gland of gastropod exposure then gradually decrease till the end molluscs is the key organof metabolism of experiment. Increments in the activities of serving also as the main site of accumulation detoxification enzymes (αand β-esterases) and biotransformation of xenobiotics may result from the stress occurred on (Desouky, 2006). Histological and enzyme expression system for synthesizing ultrastructural investigations on the digestive new and higher amounts of detoxification gland of M. cartusiana confirmed the enzymes (Wheeler and Isman, 2000). presence of three main cell types; digestive, Thisresultwas in agreement with the calcium and excretory cells forming the findings of El-Gendy (2015) who reported epithelial wall of digestive tubules. This an elevated levelof α- esterase and decreased finding was in harmony with the finding of β-esterase levels of aphids treated with Heibaet al. (2002); Ismail et al. (2013) and sublethal concentrations of Origanum Sharaf et al. (2015) who reported only three vulgare extract. These results disagreed with cell types. However, Hamed et al. (2007) the findings of Abd-El-Aziz (2014) who reported four different cell types. proved that the activity of α- esterase The present histopathological enzyme was inhibited while β-esterase examinations of the digestive gland treated activity was stimulated after treatment with with LC25 Ginger extract showed tubular Indoxacarb, Emamectin benzoate and deteriorations, tissue exudates, hemocytes Pyridalyl. Phenoloxidase is an antioxidant infiltration, excessive luminal secretions,

111 Control of the glassy clover snails Monacha cartusiana using Zingiber officinale extract as an ecofriendly molluscicide cell lysis and some necrotic areas Ginger extract was showed highly toxical appearinside tubules. Acini of ovotestis were degrees against the clover snails M. cartusiana and distorted and contained deformed egg and resulted in various biochemical, histopathological sperms. These findings were in agreement and ultrastructural disturbances. Ginger extract with Abdel-Haleem, (2014) who reported a caused severe deformations of both eggs and marked inhibition of the various stages of spermsand this may inhibit the flourishing of gametogenesis of terrestrial slugs treated clover snail's populations at Sharkia with Origanum syriacum. Governorate.Therefore, this natural product may Electronmicroscopic examinations be incorporated into apply the control programs of showed severely ruptured microvilli, land snailsinstead of using harmful synthetic lacerated cell membranes, disorganized RER pesticides. Poisonous baits control technique used and condensation of the matrix of in this study is safe for non-target organisms and mitochondria. These damaged cellular considered as asubstitutional method in spite of the alterations may be due to the direct toxic classical spray control methods. effect of Ginger extract. Mitochondrial shrinking might be due to toxicant-induced REFERENCES inhibition of Na+/H+ transporter and Abdel-Haleem, A.A. (2013). Molluscicidal impairment of the overall osmoregulatory impacts of some Egyptian plant mechanism of the cell (Vilella et al., extracts on protein and DNA- 1991).In this concern, Hamed et al. contents of two snail-vectors of (2007)showed disruption and reduction of schistosomiasis, using microvilli, increased number of calcium electrophoresis. J. Bas. & Appl. spherules and presence of large numbers of Zool., 66: 34-40. excretory granules in the excretory cells in Abdel-Haleem, A. A. (2014). Cytotoxicity the digestive gland of E. of Egyptian plant extract Origanum vermiculataexposed to methomyl. syriacum on gametogenesis of two Moreover, Sharaf El-Din et al. (2012) Egyptian terrestrial slugs, using indicated severe ultrastructural alterations in TEM. Int. J. Acad. Sci. Res., 2 (1): the cerebral ganglia of treated with 01-08. Biomphalaria alexandrina exposed to Abd-El-Aziz, H. S. (2014). Effect of some ethanolic extract of Anagalis arvensis in the insecticides on certain enzymes of form of, pyknotic nuclei, atrophy of the Spodoptera littoralis (Bosid.). Egypt. perikarya of some neurons, fragmentation or J. Agric. Res., 92 (2):501-512. dilation of rough endoplasmic reticulum, Ahmad, F.; Sagheer, M.; Hammad, A.; damage of mitochondria, and vacuolation of Rahman, S.M. and Mansoor-Ul- cytoplasm. Furthermore, Ustina et al. (2018) Hasan (2013). Insecticidal activity of investigated ultrastructural alternations in some plant extracts against the digestive gland of the Egyptian slug, Trogoderma granarium (E.). The Limax maximus exposed to thymol pesticide agriculturists, 11(1): 103-111. and they reported cytoplasmic vacuolation, Amri, A. and Touil-Boukoffa, C. (2016). In degeneration of some nuclei, rupture of vitro anti-hydatic and immuno- microvilli and increasing of calcium modulatory effects of ginger and [6]- spherules inside secretory cells. gingerol.Asia. Pac. J. Trop. Med., 9(8): 749–756. Conclusion

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115 Control of the glassy clover snails Monacha cartusiana using Zingiber officinale extract as an ecofriendly molluscicide

هكبفحه قواقع البرسين السجبجي هونبكب كبرتوزيبنب بإستخذام هستخلص السنجبيل كوبيذ رخوي صذيق للبيئه

هحوود عبذ العبطي1 ، علي الشيخ2، عبذ الونعن خليل1، وسبم الجوهري2 1- قسٌ ػيٌ اىحي٘اُ- ميئ اىؼيً٘- جبؼٍٔ اىضقبصيق ؼٍ - 2ٖذ ثح٘ث ٗقبئ اىْجبتبت– ٍشمض اىجح٘ث اىضساػئ. اىذقي. اىجيضٓ. ٍصش

الوستخلص ى٘حظت مثبفبت ػبىئ ٍِ ق٘قغ اىجشسيٌ اىضجبجي ٍّ٘بمب مبست٘صيبّب ٗاىتي تسججت في ٕالك اىؼذيذ ٍِ اىَحبصيو اإلقتصبدئ ثَحبفظٔ اىششقئ. إٔتَت ٕزٓ اىذساسٔ ثَنبفحٔ ٕزٓ األفٔ اىعبسٓ ثإستخذاً اىَستخيص اإلثييي ىْجبت اىضّججيو مَجيذ سخ٘ي غجيؼي أٍِ ػيي اىجيئٔ. ٗظحت دساسبت اىسَئ أُ اىتشميض األقو ىَستخيص اىضّججيو )%20(سجت ٗفيبت ّح٘ 10% فقػ ٍِ ق٘اقغ اىجشسيٌ ثؼذ اىيً٘ االٗه ٍِ اىؼَبٍئ ٗٗصيت ّسجٔ اى٘فيبت اىي 66.7% ثؼذ اىيً٘ اىثبٍِ ٗاىؼششيِ ٍِ اىؼَبىجٔ . سجيت أعىي ّسجٔ ٗفيبت في ق٘اقغ اىجشسيٌ ) 90%( ٗرىل ثؼذ تؼشظٖب ىيطؼً٘ اىسبٍٔ ىَستخيص اىضّججيو ثتشميض 40% ىَذٓ 28يً٘.

أظٖشت اىفح٘صبت اىجي٘ميَيبئئ أُ تؼشض اىق٘اقغ ىتشميض تحت ٍَيت ٍِ ٍستخيص اىضّججيو LC25 (%7.04) قذ أحذث إستفبػب ؼٍْ٘يب في ٍست٘ي إّضيَبت اسجشتيت اٍيْ٘ تشاّسفيشيض ٗ أالّيِ اٍيْ٘ تشاّسفيشيض ٗاىفب ٗثيتب إستيشيض ٗ اىفيْ٘ه أٗمسيذيض في اىق٘اقغ اىؼَبىجٔ ٍقبسّٔ ثبىق٘اقغ غيش اىؼَبىجٔ. مَب أٗظحت اىفح٘صبت

اىْسيجئ ىنو ٍِ اىغذٓ اىٖبظَٔ ٗاىغذٓ اىخْث٘ئ ثؼذ تؼشظٖب ىتشميض تحت ٍَيت LC25 ٍِ اىَستخيص اىؼذيذ ٍِ اإلختالالت اىتشميجئ في األّيجيجبت اىٖبظَٔ ٗإستشبحبت إىتٖبثئ ىنشيبت اىذً ٗظٖ٘س إفشاصات مثيفٔ ثذاخو تجبٗيت األّيجيجبت اىٖبظَٔ مَب ظٔست ثؼط اىَسبحبت اىْسيجئ اىَتحيئ. تشٕ٘ت اىجي٘ة اىخْث٘ئ ٗ إشتَيت ػيي خاليب ثيعئ غيش غجيؼئ ٗحي٘اّبت ٍْ٘ئ ٍشٕ٘ٔ.