Evaluation the efficacy of anthrax vaccine against challenge with a highly virulent strain of bacillus anthracis isolated from soil in sheep, goats and guinea pigs in iran
Archives of Razi Institute, Vol. 64, No. 1, June (2009) 45-50
Razi Vaccine & Serum Research Institute
In vivo effect of albendazole and mebendazole on hydatid cyst of mice Hashemi Tabar 1, 2, G.R., Razmi1, G.R., maleki1, M. 1. Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad , Mashhad, Iran 2. The Research Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran
Received 12 Oct 2008; accepted 28 Apr 2009
ABSTRACT
In current study, a few sheep cystic livers and lungs were obtained from Mashhad slaughter house and protoscolex were separated aseptically. Thirty 6-week-old Swiss mice were divided into 5 groups and two thousands of live protoscolices were inoculated intrapritoaneally in each mouse. In group 1 and 2 (prophylactic groups), mice were given 150 mg/kg of oral albendazole and mebendazole for 10 days. In group 3 and 4 (treated groups), mice were treated orally with 300 mg/kg of albendazole and mebendazole for 22 days with an interval of 2 days after every 4 days of treatment 6 months after inoculation. The control group (group 5), was sham injected with normal saline. Mice were killed after 7 months and internal organs were observed for hydatid cyst. In group 1, 2 cysts were observed in the liver of mice. In this group, although albendazole did not prevent cyst formation, but the number and also the size of the cysts were lower and smaller than the control group. In group 2, 3 cysts were observed in internal organs. In group 3, there was no cyst in internal organs and so in this group it is concluded that albendazole prevented cyst formation. In group 4, 1 cyst was observed on the liver of mice. In control group a lot of cysts were observed in internal organs of mice and the average size of cysts were bigger than prophylactic and also treatment groups. Keywords: Hydatid cyst, Albendazole, Mebendazole, Mouse INTRODUCTION
methylcarbamates derivatives (BZD) were proved
to be effective against E. granulosus, and since
Cystic echinococcosis (CE) caused by the
then, many investigators have used albendazole
metacestode (larval) stage of Echinococcus
(ABZ) and mebendazole (MBZ) for the treatment
granulosus (E. granulosus ) is still an important
of human hydatidosis (Teggi et al 1993).
public health concern in many countries of the
Albendazole and mebendazole are considered to be
world, such as Mediterranean and South American
equally effective. When evaluated up to 12 months
countries The disease affects humans as well as
after initiation of benzimidazole treatment, 10%–
domestic livestock including cattle, sheep, camels,
30% of patients exhibit cyst disappearance, 50%–
pigs, horses and others (Torgerson & Budke 2003).
70% shows degeneration of cysts and 20%–30%
In the early 1970s, benzimidazole show no morphological changes in the cyst
appearance. Continuous or intermittent treatment
*Author for correspondence. E-mail: hashemit@um.ac.ir 46 Hashemi Tabar, et al / Archives of Razi Institute, Vol. 64, No. 1, June (2009) 45-50
with albendazole is recommended for a period of 6
collected aseptically from cysts and were
maintained in Hank’s Balanced Salt Solution
benzimidazole methylcarbamate compounds such
(HBSS) containing penicillin (500 IU/ml),
as albendazole and mebendazole have been used to
streptomycin (500 IU/ml). Prior to inoculation, the
treat CE in humans (El-On. 2003). Mebendazole
collected protoscolices were rinsed 2-3 times with
was the first compound widely used (Bekhti et al
HBSS containing antibiotics. Viability of the
1997). Although effective, the main disadvantage
protoscolices was confirmed by visual inspection
was the extremely high doses and prolonged
through inverted microscope. The protoscolices
administration period required to achieve a
satisfactory clinical efficacy. Albendazole was
protoscolices in 0.5 ml of HBSS were inoculated
therefore a major breakthrough because, while
intraperitoneally in each mouse. Male and female
absorption of the parent compound remained poor,
mice weighing 20–22 g were used in these studies.
its metabolite, albendazole-sulphoxide (ABZSO)
Thirty mice were inoculated with protoscolices
was shown to be an active anthelmintic agent
of E. granulosus and divided into 5 groups. Two
(Horton. 2003). Both ABZ and ABZSO have been
prophylactic groups, just after challenge with
shown to be active against protoscolices of E.
protoscolices, received 150 mg/kg BW /day
granulosus in in vitro culture systems. However,
albendazole (group 1) and/or mebendazole (group
the in vitro protoscolicidal action of ABZ and
2) orally for 10 days. Two other groups, as
ABZSO is very slow and requires a longer
treatment groups, 6 months after challenge with
incubation period compared to that observed for
protoscolices were treated orally with 300 mg/kg of
MBZ (Pérez-Serrano et al., 1994). In vitro and in
albendazole (group 3) and mebendazole (group 4)
vivo differences in the protoscolicidal activity of
every 4 days with an interval of 2 days for totally of
BZD anthelmintics have been observed. The
24 treatment days (six times). The fifth group
significance of the effects of albendazole,
animals were assigned as control and mice were
sham injected with normal saline. All mice were
researchers (Pérez-Serrano et al 2001, Rodríguez-
sacrificed after 7 months and internal organs were
Caabeiro et alet al 2004). The aim
observed for hydatid cyst. For histopathological
of the present work was to determine in vivo
study, tissue samples were taken from liver of
protoscolicidal effect of albendazole and
control and test mice. Samples were fixed in buffer
mebendazole against E. granulosus in mouse.
formalin 10% and then histopathologic section was
prepared by routine method. Samples were stained
by hematoxylin and eosin and finally were
MATERIALS AND METHODS E. granulosus hydatid cysts containing
conditions from infected sheep with hydatid cysts
presented for routine slaughter in abattoirs in
All the mice were killed 7 months after the
Mashhad. This study has been conducted in 2008.
inoculation and internal organs were inspected for
Briefly, the hydatid cysts (2–5 cm in diameter)
hydatid cyst(s). In group 1, which received
were cut open and vesicle fluid (containing
albendazole for 10 days, 2 cysts were observed on
protoscolices) was separated from the metacestode
liver of mice. The size of cysts of this group was
tissue and host adventitia. Protoscolices were
smaller than the size of the cysts in internal organs
Hashemi Tabar, et al / Archives of Razi Institute, Vol. 64, No. 1, December (2009) 45-50 47
of the control group. In group 2, which received
the internal organs of all mice and the average size
mebendazole for 10 days, 3 cysts were observed in
of cysts were about 6.5mm and bigger than
internal organs and the size of cysts was bigger than
prophylactic and treated animals with both drugs.
the group 1, but smaller than the control group. In
Histopathological study of the cyst showed that
group 3, which treated with albendazole for 22
typical double layered wall and in some cases with
days, no cyst was found in the internal organs
clear fluid inside (Figure 2). Some samples
revealed a precystic structure, which consisted of
connective tissue and scattered inflammatory cells.
DISCUSSION
In the present study, the efficacy of albendazole
and mebendazole in the prevention of hydatid cyst
in mice was conducted. In a study by (Keshmiri et al., 2001) the effect of albendazole on hydatid cysts
of human has been evaluated. Twenty-nine patients
with 240 cysts received albendazole (400 mg twice
a day, in 3 cycles of 6 weeks long with 2 weeks off
between cycles). Some patients with liver cysts
after treatment showed increasing heterogeneity
and density suggestive of inactive cysts. Patients
with larger cysts and those with pulmonary
involvement were better responders. The observed
results are encouraging, showing high efficacy of
albendazole for the treatment of hydatidosis and
should be offered to patients as an alternative
before surgical treatment is considered. In
comparison with the results of (Keshmiri et al
2001), our results showed that when mice were
treated with albendazole, no cysts were found in
internal organs and this result indicated that
2001, & 2004, et al 2006). It
Figure 1. Treated group with albendazole. None of mice had
has been reported that continuous long term
viable hydatid cysts in internal organs (Above). Hydatid cysts in internal organs of mice in control group (Below).
Group 4, animals which were treated with
parasiticidal against larval stage of E. multilocularis
mebendazole for 24 days, just one cyst was
especially in the early stages of infection (Liu et al
observed on the liver of mice. In control group
1998). The effectiveness of albendazole (ABZ) and
(Figure 1) a high number of cysts were observed in
albendazole sulphoxide (ABZ.SO) and ABZ +
48 Hashemi Tabar, et al / Archives of Razi Institute, Vol. 64, No. 1, June (2009) 45-50
drugs (mebendazole, albendazole & ricobendazole)
Echinococcus granulosus protoscolices has been
were compared by analyzing their in vivo activity
described. The results indicated that ABZ and ABZ
against Echinococcus granulosus cysts in a mouse
+ ABZ.SO had an important effect upon larval
growth in E. granulosus (Perez-Serrano et al.,
mebendazole-treated mice, at doses of 25-50
1997). The efficacy of Ivermectin (IVM) alone,
albendazole (ABZ) alone and a combination of
IVM plus ABZ against Echinococcus granulosus
benzoimidazole used, has some beneficial effects
protoscolices was studied by means of an in vitro
on the disease in selected patients, it has also been
incubation. The maximum protoscolicidal effect
associated with treatment failure in some cases,
was detected when combination of IVM+ABZ
perhaps because of its poor absorption. It has
were used (Casado et al 2002). It has been reported
shown by et al 1998) that hydatid cysts
that protoscolices of E. granulosus were incubated
were severely damaged in mice treated with
in vitro with praziquantel (PZ), albendazole (ABZ),
mebendazole and new vesicles did not develop
or a combination of both (PZ + ABZ). PZ and ABZ
displayed slower protoscolicidal activity when
applied separately than when used in combination.
The PZ + ABZ treatment was effective only against
small cysts, which had collapsed at 10 days
postinoculation (Urrea-Paris et al 2000).
It has been reported by (Dvoroznakova et al
2004), that the reduction of cyst growth after
treatment with ABZ and liposome.ABZ was similar
up to week 4 after last dose, but the parasitostatic
effect of liposome. ABZ lasted 4 weeks longer than
the effect of free drug. At the present time,
albendazole is considered the treatment of choice
Figure 2. Microscopic cyst in liver of mice. The arrows indicate microscopic cyst and tissue reactions around cyst.
for echinococcosis in human (Shuhua et al 2002).
Feng et al (1995) have shown that when infected
Also, hydatid cysts reappeared after treatment with
mebendazole, for 14 days, the glutathione S-
transferase (GST) activity of both collapsed and full
indicated that the postoperative prophylactic course
cyst walls were inhibited by 30.1% and 26.8%,
of mebendazole is reliable, safe and with minor side
respectively. Albendazole (300 mg/kg/day) for 14
effects and the recurrence rate of the disease was
days had no apparent effect on GST of E. granulosus cyst wall and it has been suggested that
the inhibition of GST activity in the cyst wall
induced by mebendazole might damage the defense
Acknowledgment
In another experiment by (Daniel-Mwuambete et
The authors thanks to Dr. Fallah-Rad, Mr. Azari
al 2003), two different preparations, solution and
and Mr. Mohammadnejad for their time and help.
suspension, of three benzimidazole carbamate
This research was supported by the Ferdowsi
Hashemi Tabar, et al / Archives of Razi Institute, Vol. 64, No. 1, December (2009) 45-50 49
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Echinococcosis--an international public health
Glycopeptide MICs are Higher in MSSA compared with MRSA from Blood in the UK and Ireland 2001-2009 rosy.reynolds@nbt.nhs.uk R. Reynolds1, R. Hope2 and The BSAC Extended Working Party on Resistance Surveillance1 1British Society for Antimicrobial Chemotherapy, Birmingham, B1 3NJ 2Health Protection Agency, London, NW9 5HT BACKGROUND MSSA infections, best treated with a -lactam, may