Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
AENSI Journals
Advances in Environmental Biology
ISSN-1995-0756
EISSN-1998-1066
Journal home page: http://www.aensiweb.com/AEB/
In vitro, the Biological control of adult Monizia spp. by
brown algae (Fucus vesiculosus) on sheep in Taif
Governorate, K.S.A.
Nabila, S. Degheidy and Al-Otaibi, Wafa
Biology Department, Science Collage, Taif University, KSA.
Address For Correspondence:
Nabila, S. Degheidy, Biology Department, Science Collage, Taif University, KSA.,
E-mail: [email protected]
This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/
Received 12 August 2016; Accepted 17 December 2016; Available online 31 December 2016
ABSTRACT
The present study was designed for study of efficacy of watery extracts of brown algae (Fucus vesiculosus) on sheep as anti-parasitic on the
adult worms of Monizia spp. through investigate the parasite morphological changes by using Scanning Electron Microscopy (SEM). For
biological control of this parasite, different concentrations of Fucus vesiculosus (10 μg/ml; 20 μg/ml and 30 μg/ml ) were used. In vitro, the
results indicated that the Fucus vesiculosusis efficacious at all concentrations tested after 24 h specially (30μg/ml). Electron microscopic
examination showed that many structures of the treated worms were affected . The most affected sites were the scolex and the tegumental
surface and proglottides.
KEYWORDS: Biological control; monizia spp.; brown algae; Fucus vesiculosus; In vitro; SEM and sheep.
INTRODUCTION
Bladderwrack (formal name of Fucus Vesiculosis) is a brown seaweed which is a good source of iodine (the
mineral needed for proper thyroid function). And generally being anti-obesity, anti-inflammatory, anti-oxidant,
and anti-carcinogenic. There are also some implications of them being anti-viral and anti-diabetic [1].
Among livestock animals; sheep and goats are of high economic importance. Also, they are multi-purposes
animals not only for mutton and milk production but also for their medical, wool and skin. These animals arc
economic and have the ability to survive on a variety of vegetation and seeds. Furthermore, sheep and goats are
easily bred and become adapted to rural and urban environmental conditions [2] and [3]. At the same time,
helminths are the most destructive internal parasites responsible for the increased mortality and decreased
animal production as they could cause many pathological changes which may lead to severe illness or death of
the host [4]. Tape worms, which could be considered as one of the most important parasites of sheep and
monieziasis, is an important problem in sheep breeding [5;6;7]. Moreover, control of helminth parasites using
chemical products resulted environment pollution and produced resistance of parasites. Thus it is necessary to
search for other safe and potent against parasites [8;9]. Therefore, the use of biological materials instead of
chemicals in the control of parasites was documented in the last few years. It is proven that the naturally
occurring products in plant extracts have potency against different pathogens and are mostly safe in use and
non-expensive biological control [10;11;12;13;14;15;16]. Some Scientist [17] and [18] reported that Culex
pipiens mosquito larvae treated with brown algae were significantly affected and didn’t reach the pupa stage due
to the levels of protective polyphenolic compounds of brown algae’s which important chemical defense
significantly due to reducing palatability in response to direct mosquito attacks. Many species of marine algae’s
Copyright © 2016 by authors and Copyright, American-Eurasian Network for Scientific Information (AENSI Publication).
199
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
including Fucus vesiculosus have been reported to be used as bio-fertilizers due to the presence of trace
elements and in particular the growth hormone like substances such as cytokinins in them. Monica and
Schoenwaelder, 2008 [19] in USA stated that phenolics are crucial for many important aspects of plant life.
They can play structural roles in different supporting or protective tissues, for example in cell walls, they can be
involved in defence strategies, and signalling properties particularly in the interactions between plants and their
environment. In brown algae, phenolic compounds are contained within membrane bound vesicles known as
physodes and their roles in algae are thought to be similar to those of higher plant phenolics. They can be
stained using various histochemical stains, however, none of these stains are phenolic specific, so care must be
taken during interpretation of such results. Jasmine Spavieri [20], in the continuation of a research on seaweeds,
crude extracts of 21 brown algae collected from the south coast of England and the west coast of Ireland were
screened for in vitro trypanocidal ; leishmanicidal and antimycobacterial activities. Mammalian stages of a small
set of parasitic protozoa; i.e Trypanosoma brucei and rhodesiense; T. cruzi and Leishmania donovani and the
tubercle bacillus Mycobacterium tuberculosis were used as test organisms. Only four extracts were moderately
active against T. cruzi, whereas all algal extracts showed significant activity against T. brucei rhodesiense, with
Halidrys siliquosa and Bifurcaria bifurcata (Sargassaceae). Aly and W.L. Abdou, 2010 [21] studied the effect
of native Spirulina platensis on the developmental biology of Spodoptera littoralis, it can be concluded that
application of 5% concentration of water solution of the green algae, Spirulina platensis, can protect the host
plants from being attacked by S. Littoralis with the other means of integrated pest management. Screening of
some seaweeds species from South Sinai, Red Sea as potential bioinsecticides against mosquito larvae; Culex
pipiens [22]. Elbanna1 and Hegazi [22] studied on searching for bioinsecticides virulent to mosquito larvae;
Culex pipiens, nine seaweeds species were screened. In this study, different dried seaweeds were applied into
the mosquito larvae rearing jars. The effect of these seaweeds on the survival larval development and growth
rate of the mosquito Culex pipiens larvae was measured. The percentage of dead larvae fed with seaweeds
species after 10 and 15 days were 50%, 90% respectively. Delayed pupation and body size reduction of the
mosquitoes larvae fed with seaweeds were observed. The study showed also that Caulerpa prolifera; Caulerpa
serrulata; Jania rubens; Nitophyllum punctatum; Cystoseira myrica and Padina pavonica have potential to be
used as larvicidal agents acting mainly to increase life cycle intervals and pupation delay.
Bashtar et al.2012 [23] examined 2.343 sheep through the wholey year, 271 were found to be infected with
tapeworms (11.5%). The highest infection rate recorded was Moniezia expansa (74%). For biological control of
this parasite, different concentrations of crude plant extract of Artemisia cina were used in vitro and in vivo. In
vitro, there results indicated that the plant extract is efficacious at all concentrations tested. Electron microscopic
examination showed that many structures of the treated worms were affected. The most affected sites were the
scolex and the microtriches of the outer tegumental surface. Maissa, M. Morsi, et al.2016 [24] stated that
Spirulina platensis is a blue- green micro alga or cyanobacterium found in warm water and alkaline volcanic
lakes.The blue- green alga has a soft cell wall made of complex sugars and proteins. Recent studies have
demonstrated that in the microalga Spirulina platensis a blue protein called phycocyanin, belonging to the
photosynthetic apparatus has antioxidant and radical scavenging properties both in vivo and in vitro models.
Benedetti, et al. 2006 [25] and [21] recorded that application of 5% concentration of water solution of the blue
green alga, Spirulina platensis, can protect the host plants from being attacked by Spodoptera littoralis with the
other means of integrated pest management. Maissa, M. Morsi, et al.2016 [24], stated that the unwise usage of
chemical drugs for treatment of animals may pollute the soil; air and irrigation canals kill the natural predators,
negatively affect man health and some may breakdown into toxic derivatives and causes phytotoxicity problems
in sensitive crops due to the residual property. Therefore, the search for new safer types of insectcides less
harmful and biodegradable to nontoxic products have recently attracted the attention of many scientists all over
the world. Brown algae Fucus vesiculosus commonly known as: Bladderwrack; Sea kelp or Rock wrack, is a
member of the Fucaceae family, a genus of marine brown algae found on rocky seashores of the temperate areas
of the Atlantic and Pacific oceans. Teas et al.2004 [17] and Elbanna & Hegazi [22] found that Culex pipiens
mosquito larvae treated with brown algae were significantly affected and didn’t reach the pupa stage due to the
levels of protective polyphenolic compounds of brown algae’s important chemical defense significantly
reducing palatability in response to direct mosquito attacks. Many species of marine algae including Fucus
vesiculosus have been reported to be used as bio-fertilizers due to the presence of trace elements and in
particular the growth hormone like substances such ascytokinins in them; they are used as foliar spray,
application to soil, for soaking of seeds before sowing, enhancing the germination of seeds, increasing uptake of
plant nutrients, giving resistance to frost and fungal .Therefore the present study aimed to evaluate in vitro ,
some studies on efficacy of watery extracts of brown alga( Fucus vesiculosus) as antiparazitic of different
species of Monizia spp. in sheep in Taif Governorate.
200
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
MATERIALS AND METHODS
The present study was designed for , in vitro, determinationthe antiparasitic efficacy of Fucus vesiculolus
on the adult of different species of Monizia through investigate the parasite morphological changes by using
Scanning Electron Microscopy (SEM).
Identification of Monizia spp:
The adult worms were randomly collected from small intestine of sheep, which had active infection, for
identification were done according to Abd Rahman Bashtar et al. [23] and Soulsby [26] by using a light
microscope.
Source of algae:
Fucus vesiculolus brown algae were purchased from a local market in Cairo. Dry Fucus vesiculolus tablets
was powdered by electrical grinder. Three concentration (0.10, 20 and 0.30 μg/ml) were used to study the effect
of brown alga on the adult of monizia spp. One worms were examined for each concentration [21].
Samples for scanning electron microscope:
Adult worms of Monizia spp. recovered from the small intestine of slaughtered sheep at Taif abattoir. Under
sterile conditions in a laminar flow cabinet, worms were washed in warm physiological saline (37.8˚C) and put
in sterile complete RPMI 1640 culture medium containing antibiotics (penicillin, 50 IU/ml; streptomycin, 50
μg/ml). The adult of different species of Monizia spp. were subsequently transferred to fresh culture medium
containing 50% (v/v) heat denatured rabbit serum, 2% (v/v) rabbit red blood cells; as recommended by Ibarra &
Jenkins [28] and of watery extracts of (Fucus vesiculosus )at 3 different concentrations 10; 20 and 30 μg/ml..
The whole worms incubated for 24 hrs at 37.8˚C in an atmosphere of 5% CO2. The control worms incubated for
24 hrs in RPMI 1640 culture medium containing 0.1% (v/v) DMSO. Normal control worms fixed immediately
following the initial washing and one fluke were examined for each concentration, (23). Examinations were
made according to method proposed by [29] and [28] using a computer image analysis system (ELICA QWin
500, Cambridge, England). The morphological studies included scolex as well as area of tegumental of the
wormes and sucker.
RESULTS AND DISCUSSION
The difficulties associated with chemotherapy of parasitic diseases as the ineffectiveness of drug,
incomplete elimination of certain disease or parasite, and it have serious disadvantages in some developing
countries, in addition to cost, risk of misuse leading to drug resistance, environmental pollution and food
residues. Thus it is not surprising to use some biological materials in controlling parasitic development. The
hope to produce anthelminthic compounds is a major focus in parasitology today. The efficacy of plant extracts
for their antiparasitic activity has been judged on the basis of the loss of spontaneous movement and/or complete
destruction of the worm in vitro [9;30;31;32;33]. Much plant for biological control of the parasite extracts were
used before as anthelminthic components and showed various degrees of efficacy [34;35;36;37;38;39].
The present study demonstrated the effect of water Fucus vesiculolus on monizia spp. worms as
antiparazites. Scanning electron microscopic observations could be used to determine, for the first time, the
target of Fucus vesiculolus as morphological changes could be observed. These changes were concentration
dependent and consisted of destructive alterations and deformity in the cuticle of the tegumental architecture of
M. expansa. The cuticle of the parazites is metabolically active and morphologically specialized for selective
absorption of nutrients and osmoregulation. Thus, passive diffusion of anthelmintic through the cuticle [21]
would probably be responsible for destructive changes and deformation of the parazite body surface [17]. In
cestodes, the general body surface acts as an absorptive surface according to[40]. In the present investigation,
the body surface of these helminthes was observed to be affected and altered by Fucus vesiculolus. Similar to
the present observations, the surface cuticle or tegument was found to be a principal target site for different
synthetic drugs and natural anthelmintic products as proved by histomorphological and ultrastructural studies
[41;42;28; 43]. Results in this study revealed by scanning electron micrographs (SEM) of the normal fresh
worm Moniezia spp., the scolex, sucker and strobila appeared normal and not changed (Figure a and b). After
24 h incubation with 10 μg/ml Fucus vesiculolus, the changes in adult Moniezia spp., concerned the
proglottides other than the scolex which retained its normal morphology (Figure c). The proglottides lost their
normal aspect showing wrinkled tegumental surface throughout the strobila (Figure d). On increasing the
waterly concentration to 20 μg/ml Fucus vesiculolus, the changes concerned the whole-body surface.The scolex
appeared to be more swollen than normal with contractions to the sucker’s opening (Figure e). The proglottides
appeared deformed with circular areas of pronounced swelling occurred along their margins (Figure f). With
higher concentration to 30 μg/ml Fucus vesiculolus, the adult cestodes showed shrunken scolex with severely
201
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
distorted and folded tegument around the suckers so that a complete closure of their openings had occurred
(Figure g). SEM following 24 h of incubation with 30 μg/ml Fucus vesiculosus, the proglottides of Monizia
spp.showing tegumental irregular cracks and severely distorted in different magnification (Figure h and h). In all
of the experiments, no significant differences were observed between fresh and control worms incubated for 24
h in solvent, 0.1% (v/v) DMSO. The swelling and distortion of the body surface of M. expansa induced by
Fucus vesiculolus had been described for these helminthes by Nigella sativa oil [44]. In Fasciola hepatica, a
similar pattern of tegumental swelling had been described following incubation with a number of anthelmintic,
as summarized by (45;46). This was a common feature of drug treated parasites and considered to be an
indicator of a stress reaction by the parasite, in an attempt to replace damaged surface membrane. The scientists
[45;46] and [47] showed that the brown algae Padina pavonica was the most effective species affect
significantly the larval growth rate as all mosquito larvae died at day 10 with ρ>0.0001, none of larvae treated
with Padina pavonica reached fourth larval instars, significantly different from the control ρ>0.0005; larvae
were significantly affected and didn’t reach the pupa stage.This may be due to brown algae contain high
phenolic contents and type of tannin called phlorotannins.The larvicidal effect of brown algae may be due to
also present of phenolic, terpenoids or unsaturated fatty acids. The effect of algae’s on V. faba seeds could be
explained by several authors [48] who determined that a number of cyanobacteria produce, accumulate and
liberate 3indol acetic acid. Sergeeva et al.2002 [49], recorded the activity of auxin and cytokinin by three
cyanobacterial strains. Zaccaro et al. 2010 [50], reported that, cyanobacteria produces bioactive compounds
including plant growth regulators such as naphthalene acetic acid (NAA). All these compounds have the ability
to stimulate germination and growth of plants.
Further studies with,in vivo, effect of Fucus vesiculolus on gastrointestinal helminthes are clearly
warranted. Elbanna & Hegazi 2011 [18] found that, when 2nd instar Culex pipiens mosquito larvae fed on dried
marine algae, the larvae neither die nor develop normally; groups of larvae that received dried algae survived
but they didn’t go through the normal development in comparison with the control group, significantly different
from the control ρ>0.0005; the study also showed that the brown algae Padina pavonica was the most effective
species affect significantly the larvae growth rate as all mosquito larvae died at day 10 with ρ>0.0001, none of
larvae treated with Padina pavonica reached fourth larval instars, significantly different from the control
ρ>0.0005; larvae were significantly affected and didn’t reach the pupa stage this may be due to brown algae
contain high phenolic contents.
Zubia et al. 2008[51] studied phenolic compounds and biological activities of two range-extending brown
algae, they reported that brown algae produce a specific type of tannin called phlorotannins, phenolic,
terpenoids or unsaturated fatty acids which responsibale for larvicidal effect. Maissa, M. Morsi et al.2016 [24]
reported that, the effect of three bio-components (S. platensis; F. vesiculosus and Radiant SC12%) on the
mortality of adult B. incarnates. Statistical analysis revealed highly significant differences between three
components data investigated that the percentage mortality of F. vesiculolus reached 100% after 14 days.
Our results are in agreement with those of previously reported by [24;23;16;37;9;38;39; 41 and 38].
Fig.(a,b): SEM of the anterior end of adult Moniezia expansa of normal fresh worm showing a globular scolex
with four oval suckers and an elongated ribbon-like structure called strobila.
Fig. (c, d): SEM following 24 h of incubation with 10 μg/ml Fucus vesiculosus, showing wrinkled tegumental
surface throughout the strobila.
202
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
Fig.(e, f): SEM following 24 h of incubation with 20 μg/ml Fucus vesiculosus. The scolex becomes more
swollen than normal with narrowing of sucker’s opening. The proglottides appear deformed with
circular areas of pronounced swelling occurred along their margins.
g
Fig. (g): SEM following 24 h of incubation with 30 μg/ml Fucus vesiculosus, showing shrunken and corrugated
scolex with severely distorted and folded tegument.
h
203
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
h
Fig. (h and h): SEM following 24 h of incubation with 30 μg/ml Fucus vesiculosus, showing Monizia
spp.showing tegumental irregular cracks (arrows)
In conclusion, concerning the anthelmintic effect of waterly extraction of brown algae (Fucus vesiculolus)in
the treatment of Monizia expansa, it had antiparazitic activity against tape worms among sheep., less expensive
,easy used and had no side effect. Further studies are required to validate our findings and improve our
knowledge on the potential of the Fucus vesiculolus extract as antiparazitic drugs.
REFERENCES
[1] Abeer, A. Abu Zaid, Doaa M. Hammad and Eman M. Sharaf, 2015. Antitoxidant and Anticancer of
Spirulina platensis water Extracts.Asian . Network for Scientific Information.
[2] Arvinder, K., 1995. Epizootiology and biology of common anoplocephaline cestodes in sheep along with
histopathology and haematology of the definitive host. J Vet Parasitol, 9: 153-154.
[3] Repossi, P.F., M.P. Barcellos, L.O. Trivilin, I.V. Martins, P.C. da Silva, 2006. Prevalence and control of
gastrointestinal parasitosis in calves from dairy farms in the municipality of Alegre Espirito Santo. Rev
Bras Parasitol., 15(4): 147-150.
[4] Soulsby, E.J.L., 1986. Text book of: helminths, arthropods, and protozoa of domesticated animals. 7th edn.
The English Language Book Society and Bailliere Tindall, Loncon.
[5] Becker, B., H. Mehlhorn, P. Andrews and H. Thomas, 1981. Ultrastructural investigations on the effect of
Praziquantel on the tegument of five species of cestodes. Z Parasitenkd, 64(3): 257-269.
[6] Pale, P., V. Tandon, 1998. Anthelmintic efficacy of Flemingia vestita (Fabaceae): genistein-induced
alterations in the esterase activityin the cestode, Raillietina echinobothrida. J Biosci, 23: 25-31.
[7] Maziad, SA. and H.I. El-Nemr, 2002. The endoparasites of sheep and goats and shepherd in North Sinai
Governorate, Egypt. J Egypt SocParasitol., 32(1): 119-126.
[8] Bogh, H.O., J. Andreassen, J. Lemmich, 1996. Anthelmintic usage ofextracts of Embelia schimperi from
Tanzania. J Ethnopharmacol., 50: 35-42.
[9] Haridy, F.M., H.A. Dawood, T.A. Morsy, 2004. Efficacy of Commiphora molmol (Mirazid) against sheep
naturally infected with Monieziasis expansa in Al-Santa Center, Gharbia Governorate. Egypt J Egypt Soc
Parasitol., 34(3): 775-782.
[10] Nassar, M.I., O.M. Kandil, M. Hassanain, 1997. The potentiality of Sorghum bicolar and Nerium oleander
extracts in the control of the grey flesh fly Parasarcophaga angyrostoma (Diptera:Sarcophagidae). J Egypt
Ger Soc Zool., 22D: 235-251
[11] Abdel-Ghaffar, F., M. Semmler, 2007. Efficacy of neem seed extract shampoo on head lice of naturally
infected human in Egypt. Parasitol Res., 100: 329-332.
[12] Mehlhorn, H., 2008. Encyclopedia of Parasitology, 3rd edn. Springer Verlag, Heidelber
[13] Cenci, F.B., H. Louvandini, C.M. McManus, A. Dell’Porto, D.M. Costa, S.C. Araujo, A.P. Minho, A.L.
Abdalla, 2007.Effects of condensed tannin from Acacia mearnsii on sheep infected naturally with
gastrointestinal helminthes. Vet Parasitol., 144(1–2): 132-137.
[14] Abdel-Ghaffar, F., M. Semmler, K. Al-Rasheid, H. Mehlhorn, 2009. In vitro efficacy of byemite and mite
stop on developmental stages of the red chicken mite dermanyssus gallinae. Parasitol Res., 105(5): 14691471.
[15] Abdel-Ghaffar, F., M. Semmler, K. AL-Rasheid, S. Klimpel, H. Mehlhorn, 2010a. Comparative in vitro
tests on the efficacy and safety of 13 anti-head-lice products. Parasitol Res., 106: 423-429.
204
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
[16] Abdel-Ghaffar, F., M. Semmler, K. Al-Rasheid, K. Klimpel, H. Mehlhorn, 2010b. Efficacy of a grapefruit
extract on head lice: a clinical trial. Parasitol Res., 106: 445-449.
[17] Teas, J., S. Pino, A. Critchley and L.E. Braverman, 2004. Variability of iodine content in common
commercially available edible seaweeds. Thyroid., 10: 836-841.
[18] Elbanna, S.M. and M.M. Hegazi, 2011. Screening of some seaweeds species from South Sinai, Red Sea as
potential bioinsecticides against mosquito larvae; Culex pipiens. Egypt. Acad. J. biolog. Sci., 2: 21-30.
[19] 1Monica, E.A. Schoenwaelder, 2008. The Biology of Phenolic Containing Vesicles. Algae. 23(3): 163175.
[20] Jasmine, Spavieri, 2015. Antimycobacterial, Antiprotozoal And Cytotoxic Potential Of Twenty-One
Brown Algae (Phaeophyceae) From British And Irish Waters. British Journal Of Clinical Pharmacology
The British Pharmacological Society Special Issue: Adverse Drug Reactions, 80(4): 611-926.
[21] Aly1, M.S. and W.L. Abdou, 2010. The effect of Native Spirulina platensis on the Developmental Biology
of Spodoptera littoralis (Boisd. Journal of Genetic Engineering and Biotechnology, 8(1): 65-70.
[22] Shereen, M. Elbanna1 and Muhammad M. Hegazi, 2011. Screening of some seaweeds species from South
Sinai, Red Sea as potential bioinsecticides against mosquito larvae; Culex pipiens. Egypt. Acad. J. biolog.
Sci., 4(2): 21-30.
[23] Abdel-Rahman Bashtar and Mohey Hassanein and Fathy Abdel-Ghaffar and Khaled Al-Rasheid and Soad
Hassan and Heinz Mehlhorn and Magda AL-Mahdi and Kareem Morsy and Ali Al-Ghamdi, 2011. Studies
on monieziasis of sheep I. Prevalence and antihelminthic effects of some plant extracts, a light and electron
microscopic study. Parasitol Res., 108: 177-186.
[24] Maissa, M. Morsi, Hammad, Doaa M. and Rashwan, S. Rania, 2016. Efficiency of Three Bio-components
against broad bean beetle (Bruchidius incarnates) and Their Effect on Germination, Seedling Growth and
Cytogenetic Changes of Vicia faba L. plants. Research Journal of Pharmaceutical, Biological and
Chemical Sciences, 7(1): 1837.
[25] Benedetti, S., S. Rinalducci, F. Benvenuti Purification, 2006. and characterization of phycocyanin from the
bluegreenalga Aphanizomenonflos-aquae. Journal of Chromatography B: Analytical Technologies in the
Biomedical and Life Sciences, 1: 12-18.
[26] Soulsby, E.J.L., 1982. Text book of: helminths, arthropods, and protozoa of domesticated animals. 6th edn.
The English Language Book Society and Bailliere Tindall, Loncon
[27] Shalaby, H.A., N.M.T. Abu El Ezz, T.K. Farag and H.A.A. Abou-Zeina, 2012. In vitro efficacy of a
combination of ivermectin and Nigella sativa oil against helminth parasites. Global Veterinaria, 9: 465473.
[28] Ibarra, O.F. and D.C. Jenkins, 1984. An in vitro screen for new fasciolicidal agents.Zeitschrift
Parasitenkunde. 70: 655-661.
[29] Valero, M.A., M.D. Marcos and S. Mas-coma, 1996. Amathematical model for the ontogeny of Fasciola
hepatica in the definitive host. Research and Reviews in Parasitology, 56: 13-20.
[30] Goto, C., S. Kasuya, K. Koga, H. Ohtomo and N. Kagel, 1990. Lethal efficacy of extract from Zingiber
officinal (traditional Chinese medicine) or [6] Shogaol and [6] gingerol in Anisakis larvae invitro. Parasitol
Res., 76: 653-606.
[31] Robinson, R.D., L.A.D. Wiliams, J.F. Linda, S.I. Terzy and A. Mansingh, 1990. Inactivation of
Strongyloides stercoralis larvae in vitro by six. Parasitol Res, 108: 177: 186-185.
[32] Togo, J., M.T. Santamarina, D. Peris, F.M. Uberia, S.L. Leiro and M.L. Sanmartin, 1992. Jamaicon plants
extracts and three commercial anthelmintics. West Indian Med J. 39: 213-217.
[33] Nabila, S. Degheidy, Sharaf, Eman M. and Fathi, M. Samia, 2013. Field Evaluation of Anthelmentic
Efficacy of Calligonum comosum against Fasciolosis in Sheep at Taif, KSA. Global Veterinaria., 11(4):
377-384.
[34] Akhtar, M.S., I. Ahmad, 1992. Comparative efficacy of Mallotus philippinensis fruit (Kamala) or Nilzan
R. drug against gastrointestinal cestodes in Beetal goats. Small Ruminant Res., 8: 121-128.
[35] Saha, A., N.K. Ghosh and S.P.S. Babu, 1999. Cestocidal activity of Gladiolus gandavensis. J Parasitic Dis.,
23: 135-136.
[36] Akhtar, M.S., I. Javed, 1991. Efficacy of Nigella sativa Linn. Seeds against Moniezia infection in sheep.
Indian Vet J., 68: 726-729.
[37] Nabila, S. Degheidy, and S. Al-Malki, Jamila, 2012. Epidemiological Studies of Fasciolosis in Human
and Animals at Taif, Saudi Arabia. World Applied Sciences Journal., 19(8): 1099-1104.
[38] Jamila S. Al-Malki, 2013. In vitro Efficacy of a Combination of Stromectol and Nigellsativa Oil Against
Cestodesin Sheep, Taif, Saudi Arabia. World Applied Sciences Journal., 27(3): 413-417.
[39] Nabila, S. Degheidy, Sharaf, Eman M. and Fathi, M. Samia, 2013. Field Evaluation of Anthelmentic
Efficacy of Calligonum comosum against Fasciolosis in Sheep at Taif, KSA. Global Veterinaria., 11(4):
377-384.
205
Nabila, S. Degheidy and Al-Otaibi, Wafa 2016
Advances in Environmental Biology, 10(12) December 2016, Pages: 198-205
[40] Haiba, A.A.A., N.R. Abd El-Hamid, E.A.A. Abd El-Hady and A.M.F. Al-Ansary, 2011. Cytogenetic
effect of Insecticide Telliton and Fungicide Dithane M-45 on Meiotic Cells and Seed Storage Proteins of
Vicia faba. .Journal of American Science, 1: 152-161.
[41] Abdel-Zaher, T.A.Z., 2005. Advanced studies to use plant extracts against some insect pests. Ph.D. diss.,
Faculty of Agriculture, Zagazig University, Moshtohor, Egypt.
[42] Bindhu, K.B., 2013. Effect of Azolla Extract on Growth Performance of Pisum Sativum. Int. Res. J.
Biological Sci., 10: 88-90
[43] Marcano, L., I. Carruyo, A. Del Campo and X. Montiel, 2004. Cytotoxicity and mode of action of maleic
hydrazide in root tips of Allium cepa L. Environ. Res., 94: 221-226.
[44] Shalaby, H.A. and Farag, 2014. Body Surface Changes in Gastrointestinal Helminthes Following in
vitroTreatment with Allium sativum Oil. J Veterinar Sci Technolo, 5: 1.
[45] Adam, Z.M., E. Mikhael, Z.M. El-Ashry and N.O.R.T. Ehsan, Ali., 2014. Comparative Cytogenetic and
Ultra-Structural Effects of Storing Dusted Seeds of Vicia faba with the Insecticide “Malathion 1%” and
Two Insecticidal Active Plant Products. World Applied Sciences Journal, 7: 1423-1436.
[46] Zubia, M., C. Payri and E. Deslandesm, 2008. Alginate, mannitol, phenolic compounds and biological
activities of two range-extending brown algae, Sargassum mangarevense and Turbinaria ornata
(Phaeophyta: Fucales), from Tahiti (French Polynesia). Journal of Applied Phycology, 6: 1033-1043.
[47] Schnitzler, I., G. Pohnert, H. Hay and W. Boland, 2011. Chemical Defense of Brown Algae (Dictyopteris
spp.) against the Herbivorous Amphipod Ampithoelongimana. Oecologia. 4: 515-521.
[48] Schnitzler, I., G. Pohnert, H. Hay and W. Boland, 2001. Chemical Defense of Brown Algae (Dictyopteris
spp.) against the Herbivorous Amphipod Ampithoelongimana. 4: 515-521.
[49] Sergeeva, E., A. Liaaimer and B. Bergman, 2002. Evidence for production of the phytohormone indole
3acetic acid by cyanobacteria. Planta. Pp: 229-238.
[50] Zaccaro, M.C., A. Kato, G. Zulpa, M.M. Storni, N. Steyerthal, K. Lobasso and A.M. Stella, 2010.
Bioactivity of Scytonema hofmanni (Cyanobacteria) in Lilium alexandrae in vitro propagation. Electronic
Journal of Biotechnology, 3: 210-214.
[51] Zubia, M., C. Payri and E. Deslandesm, 2008. Alginate, mannitol, phenolic compounds and biological
activities of two range-extending brown algae, Sargassum mangarevense and Turbinaria ornata
(Phaeophyta: Fucales), from Tahiti (French Polynesia). Journal of Applied Phycology, 6: 1033-1043.