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Research Article z Available online at http://www.journalcra.com INTERNATIONAL JOURNAL OF CURRENT RESEARCH International Journal of Current Research Vol. 9, Issue, 07, pp.54892-54898, July, 2017 ISSN: 0975-833X RESEARCH ARTICLE EGGS VIABILITY AND HISTOLOGICAL CHANGES IN OVARY OF ARGAS (PERSICARGAS) PERSICUS (ACARI: ARGASIDAE) AFTER TREATMENT WITH ALLIUM SATIVUM EXTRACT Shimaa S. Ahmed, Ola H. Zyaan and *Mohamed A. Abdou Department of Entomology, Faculty of Science, Ain Shams University, Cairo, Egypt ARTICLE INFO ABSTRACT Article History: Agriculturists in developing countries are suffering from many diseases caused by tick infestations Received 22nd April, 2017 that reduce the productivity of their livestock. To diminish these losses, natural products (eco-friendly) Received in revised form for ectoparasite control with lower chance of improvement of resistance are required. This study 15th May, 2017 aimed to examine the effect of different concentrations (0.5, 1.5, 2.5, and 4%) of Allium sativum Accepted 30th June, 2017 (Garlic) extract on the viability of eggs laid by Argaspersicus females and their ovaries’ development. Published online 31st July, 2017 It was found that the number of eggs laid by treated females were significantly(P<0.05) decreased than those laid by normal females. There is a significant inverse relationship between the treatment Key words: concentration of garlic extract and the percentage of hatched eggs. Applying different concentrations (100, 200, 400, 500, and 600 ppm) of garlic extract on the eggs resulted in the percentage of Argas persicus, unhatched eggs increasing significantly as garlic concentrations also increased.The average diameters Allium sativum (Garlic) extract, of oocytes in treated females were decreased by 61% from the normal oocytes’ average diameter, and Acaricides, Ovary-histology, the ovary appeared studded with previtellogenic primary oocytes. Histological changes observed in Eggs viability, the treated ovaries include: the presence of vacuolization; alteration of oocyte morphology, which Vitellogenesis. changed from rounded to elongate; disorganization of the yolk granules; and deformation of the chorion. These results demonstrate that garlic extract affected A. persicus oogenesis by interfering with the formation of yolk granules and egg shells. Our results suggest that garlicextract causes partial blockage of the vitellogenesis and other aspects of oogenesis as indicated by disrupted synthesis of yolk protein in the newly formed oocytes. Copyright©2017, Shimaa S. Ahmed et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Citation: Shimaa S. Ahmed, Ola H. Zyaan and Mohamed A. Abdou, 2017. “Eggs viability and histological changes in ovary of Argas (persicargas) persicus (Acari: Argasidae) after treatment with Allium sativum extract”, International Journal of Current Research, 9, (07), 54892-54898. INTRODUCTION encouraging results (Mehlhorn et al., 2005; Mehlhorn, 2008; Kamaraj et al., 2010; Ghosh et al., 2011; Semmler Argas (Persicurgas) persicus (Oken, 1818) is an ectoparasite et al., 2011; Sing et al., 2013). Herbal preparation consisting of of domestic poultry and wild birds in parts of Asia, Europe, garlic extracts along with extracts of lemon and onion mixed Africa and Australia. It inhabits cracks and crevices in deserts, with turmeric and camphor powder in Pongamia(Karanj) oil forest and steppe zones near human habitations (Hoogstraal, was applied for 5days to eliminate Sarcoptesscabiei infestation 1973). It is a vector of the poultry spirochaetosisTreponema in piglets (Dwivedi and Sharma, 1986). Plant extracts like gallinarum (Burroughs, 1947) and Borrelia anserine (El- Allium plant extract have been proven to be potential kammah et al., 1982) as well as Quaranfil virus in Egypt alternatives to conventional insecticides;garlic (Allium (Kaiser, 1966; El-kammah and AdbelWahab, 1981). The great sativum) acts as an antiparasitic agent as a result of the destruction caused by these ticks to animal productivity has presence of its main component, allicim(Ankri and increased the need for their control. The rapid development of Mirelman,1999). It has larvicidal activities against filarial tick resistance against active substances in synthetic acaricides mosquito, Culexquinquefasciatus (Shrankhlaet al., 2011). It (Ghosh et al., 2006; George, 2000), together with related also has an antiprotozoal effect (Sharma et al., 2009;Yakoob environmental and health risks (Hodgson and Levi, 1996) et al., 2011). Essential oils of garlic produce high mortality(90- associated with their application, increase the need for natural 100%) on 10-day old Rhipicephalusmicroplus larva when used products as an alternate to synthetic acaricides. Compounds of in certain concentrations (Martinez-Velazquez et al.,2011). plant origin have been used recently against all stages (adults, Application of 10% garlic treated mite infestation in hens larva, and nymph) of economically important tick species with (Birrenkott et al., 2000). Treatment of Rhipicephalusmicroplus females with 0.25% Melia azedarach (plant extract) resulted in *Corresponding author: Mohamed A. Abdou, reduction in ovary weight. In addition, histological techniques Department of Entomology, Faculty of Science, Ain Shams University, Cairo, revealed morphological changes in the ovary such as Egypt 54893 Shimaa S. Ahmed et al. Eggs viability and histological changes in ovary of Argas (Persicargas) Persicus (Acari: Argasidae) after treatment with Allium sativum extract deformation of the chorion; and the yolk granules were solution. Eggs used as control were treated with appropriate disorganized (Sousa et al., 2013). Therefore, this study was amount of distilled water. They were incubated at 27°C and conducted to investigate the potency of garlic extract as an 75% RH in a laboratory until the eggs hatched to larvae after 7 acaricide on the reproductive potential and the histological - 14 days. Eggs which failed to hatch after 14 days were morphology of the female ovary of Argaspersicus. counted as unhatched eggs. MATERIALS AND METHODS Histological study Ticks Ovaries of 6-day old females from normal and treated groups were immersed in 3% glutaraldehyde then transferred to initial The soft tick, Argas (Persicargas) persicus (Oken) was fixative. They were fixed overnight and given 3 brief rinses in collected from a domestic chicken house at Beni Suef a sucrose-cacodylate buffer (pH 7.2); post fixed 1hr in 1% Governorate, Egypt. To establish a laboratory colony, ticks osmium tetroxide in 0.1M cacodylate buffer, rinsed 3 times in distilled water, and dehydrated through a graded series of were maintained at 27C 1, 75% RH and 16 hrs day light. ethanol. Then specimens were infiltrated with Spurr's epoxy The ticks were held in transparent polyethylene tubes (25 resin in a graded series of absolute alcohol-Spurr's resin 100 or 13 100 mm) which were sealed at one end by a plaster mixtures, then embedded in freshly prepared Spurr's resin at of pairs plug (rearing tube) and covered at the other end with 70°C for 72hrs. Semithin sections were stained with toluidine muslin cloth securely held by rubber bands (Kaiser, 1966). blue. Specimens were examined and photographed by Domestic pigeons (Columba livia), from a commercial breeder equipped Leica light microscope with a camera using objective in Cairo, were used for feeding the ticks. Both colony and host lens 40X and 100X with 20, 50 and 100 µm scales. Oocytes were kept in an insectary provided by the Department of were measured with the image analysis software, ImageJ Entomology, Ain Shams University. The pigeon host was tied (Rasband, 1997). to a wooden board with one wing stretched laterally. The inner wing feathers were plucked and ticks (adults and nymphs) Statistical Analysis were placed to feed on the wing for 15- 20 min. Following engorgement, ticks were transferred to a Petri dish containing a filter paper disk and observed until coxal fluid was emitted. The obtained data was subjected to analysis of variance (ANOVA) followed by post-hoc analysis (Tukey’s HSD test) with the help of SPSS version19 for Windows, in which the Allium sativumextract equation of the standard deviation, standard errors, and probabilities (p) were used. The level of significance was Garlic (Allium sativum) extract was prepared according to expressed as significant (P<0.05) and non-significant (p > Aboelhadid et al. (2013) by subjecting the fresh plant bulbs to 0.05).Probit analysis was used for the calculation of the LC50 steam distillation to obtain the extract. The obtained extract value. was purified chromatographically by HPLC according to Rembold et al. (1984) RESULTS Female treatment Effect of different concentrations of Allium sativum (Garlic) extract on viability of eggs laid by Argas persicus Different concentrations of garlic extract were topically females treated on 3rd day after feeding applied on the ventral side of the posterior half of female body. Ten microliters of the tested compound were applied to the rd rd Females on 3 day after feeding were treated with different mated female ticks on 3 day after feeding. Ten pairs of concentrations of garlic extract (0.5, 1.5, 2.5 and 4%) and kept treated females and normal males were placed separately in at 27°C and 75% RH. The eggs laid by treated female ticks glass vials covered with muslin cloth and were
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