الفهرس 
Flusilazole metabolismOnly 14 pages are availabe for public view
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Abstract
The present study was designed to investigate the toxic effect of
the triazole antifungal agent, flusilazole, by sub-chronic toxicity study at
doses 86.748 and 173.496 mg/kg b.w. which represented 1/20 &1/10 of
flusilazole LD50 respectively for consecutive five days per week for 90
days. As well as examine the possible protective role of aqueous
extracts of the edible portion of date fruits (Phoneix dactylifera L.) at
doses of 6 and 12 g/kg b.w. against the haematotoxicity and
hepatotoxicity induced by flusilazole, beside the oxidative stress, lipid
profile, thyroid hormones levels and residual analysis of tested fungicide
in plasma and liver tissues.
Acute oral toxicity study
An acute oral toxicity test was conducted with adult male albino
rats to evaluate the potential for flusilazole formulation to produce
toxicity from a single dose via the oral route. The median lethal dose
(LD50) of flusilazole fungicide formulation (40% EC) after a single oral
administration was found to be 1734.96 mg/kg body weight.
Sub-chronic toxicity study
The present investigation was conducted on healthy adult male
albino rats weighed about160±10g. Dose selection for Sub-chronic
toxicity study was based on the median lethal dose (LD50) of flusilazole.
Rats were divided to equal nine groups (10 rats each). The 1st group as
Control, the 2nd and 3rd groups orally treated with 86.748 and 173.496
mg/kg b.w. which represented 1/20 &1/10 of flusilazole LD50
respectively. The 4th and 5th groups orally received 6 and 12 g/kg b.w. of
aqueous date extract respectively. The 6th and 7th groups supplemented
orally with low dose of aqueous date extract before treated with 1/20
and 1/10 of flusilazole for 2 hours. The 8th and 9th groups supplemented
orally with high dose of aqueous date extract before administered with
1/20 and 1/10 of flusilazole for 2 hours as a prophylactic agent for
successive five days per week for 90 days.The blood samples were collected from all survival animals at the
end of the experiment for determination of various haematological and
different blood biochemical parameters. In addition, the assessment of
thyroid function was performed. After the last day of treatment, all the
survival rats were sacrificed and the livers were storage at -50 0C and
then homogenate for estimation of malondialdehyde (MDA), reduced
glutathione (GSH) levels, Gamma-glutamyl transferase (GGT) and
antioxidant enzymes activities included superoxide dismutase (SOD),
catalase (CAT), glutathione peroxidase (GPx) and glutathione Stransferase
(GST). However, the residual analysis was performed in the
plasma and liver tissue at the end of the experimental period. The
present results could be summarized as follow:
Haematological measurements
The haematological parameters including total erythrocyte count
(RBCs), haemoglobin concentration (Hb), packed cell volume (PCV),
Wintrobe erythrocyte indices i.e. mean corpuscular volume (MCV),
mean corpuscular haemoglobin (MCH) and mean corpuscular
haemoglobin concentration (MCHC) as well as reticulocytes count,
Hienz bodies detection and osmotic fragility of erythrocytes (OF)
However, the total leukocyte count (WBCs) and the absolute differential
leukocyte count were determined to give us vital information regarding
the status of immunity in experimental animals. The results in the
current study, revealed that treatment of rats with flusilazole at both
selected doses induced a significant increase in erythrocytic mass
accompanied by increase in haemoglobin concentration and PCV at low
dose only as well as produced a significant increase in MCH and MCHC
at high dose and a significant decrease in MCV at both selected doses.
Rats administered with the aqueous extract of date fruits alone at both
doses exhibited a significant erythrocytosis accompanied by an increase
in haemoglobin concentration while a significant decrease in MCV was
observed.
Serum hepato-specific enzymes markers (ALAT & ASAT)
The core biochemical tests recommended for liver function
ALAT and ASAT. Serum total protein and protein electrophoresis were measured to evaluate the effect of the tested fungicide on protein
synthesis. Rats administered with flusilazole at high dose induced a
significant increase in ALAT activity but showed a significant reduction
in the activity of ASAT, as well as a significant reduction in ALAT and
ASAT activities were observed at administration of low dose of
flusilazole. Oral administration of date extract at both doses either alone
or concomitant with flusilazole at both selected doses exhibited a
significant reduction in ALAT whereas, ASAT was significantly
increased.
Hepatic oxidative stress measurements
Lipid peroxidation (LPO) considered as a major contributor to the
cell function under oxidative stress conditions and it is usually indicated
by MDA level. A markedly elevation in hepatic lipid peroxidation was
observed in rats administered with low dose of the fungicide this
occurred along with a significant reduction in hepatic reduced
glutathione content. While, no changes in both parameters were
observed in rats treated with high dose of flusilazole. Rats received low
dose of date extract induced a significant reduction in MDA level
associated with a significant elevation in GSH content. Concomitant
treatment of date extracts with the tested fungicide at both doses induced
a significant elevation in MDA level accompanied by a significant rise
in hepatic reduced glutathione content.
Rats intoxicated with flusilazole at both doses induced a
significant increase in hepatic GGT activity. Also, the same trend was
noticed in rats treated with low dose of date extract either alone or
concomitant with flusilazole at two selected doses while, a significant
reduction was observed in rats treated with high dose of extract either
alone or concomitant with the fungicide at both doses.
The present results revealed that a significant decrease in the
activity of SOD was recorded whereas, GST and CAT activities
remaining unchanged in rats intoxicated with low dose of flusilazole.
Concomitant treatment of this dose with low dose of date fruit extract
ameliorated the activities of SOD, CAT and GST. On the other hand, the
group received high dose of flusilazole caused drastic alterations in antioxidant defence system causing increase in the SOD, GST and CAT
activities. Meanwhile, GPx activity was reduced significantly at both
selected doses. The results revealed that rats treated with low dose of
date extract concomitant with low dose of flusilazole exhibited
prophylactic effect of the liver from oxidative damage. Rats
administered with high dose of date extract exhibited a significant
decrease in the activities of SOD and GPx, while the activities of CAT
and GST were significantly elevated. Administration rats with high dose
of date extract concomitant with both doses of flusilazole induced a
significant reduction in the activities of SOD, CAT, GPx and GST.
Glucose level and lipid profile measurements
The glucose and lipid profile, including triacylglycerol (TAG),
total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and
high density lipoprotein-cholesterol (HDL-C) were determined to
evaluate the effect of flusilazole on glucose and lipid metabolism. A
significant increase was observed in the glucose level in all treatments
except rats administered with high dose of date extract throughout the
experimental period.
Animals were treated with the two doses of flusilazole exerted
various effects on the serum triacylglycerol, total cholesterol, LDL-C
and HDL-C levels. After the high dose of treatment TAG, TC and LDLC
were significantly increased accompanied by reduction in the HDL-C
level, while treatment with the low dose of flusilazole induced a
significant reduction in TC and HDL-C levels. In the present study,
although animals those receiving high dose date concomitant with low
or high dose of flusilazole showed a significant elevation in TAG
levels but exhibited a significant reduction in TC and LDL-C and HDLC.
Meanwhile, rats received low dose of date extract either alone or
prior to the administration with flusilazole at both selected doses caused
a significant reduction in TAG levels.Serum total protein and protein electrophoresis
High dose of flusilazole induced a significant elevation in
globulin concentration. Significant reduction in rats treated with date
extract at both doses prior to flusilazole-intoxication at low dose in total
protein and albumin concentration. While, rats administrated both doses
of extracts prior to flusilazol intoxication at high dose induced a
significant elevation in total protein, albumin, globulin, alpha, beta and
gamma globulin. Rats administrated with low dose of date extract
induced a significant elevation in globulin, beta and gamma globulin.
Thyroid hormone levels
Measurement of thyroid hormones level is performed to estimate
thyroid gland function. A hyperthyroidism (T4&T3) was observed at low
dose of flusilazole-treated rats while, no significant alterations were
noticed in high dose-treated animals. Date extracts concomitant with
flusilazole at both doses exhibited a significant reduction in
triiodothyronine levels while the level of throxine was significantly
elevated except in rats treated with a low dose of extract prior to
flusilazole intoxicated with high dose.
Residual analysis of flusilazole in plasma and liver tissue
Results showed that the residues level of flusilazole in plasma and
liver tissues were below the limit of detection (0.01μg g-1).
In conclusion, the daily oral consumption of an aqueous extract of
date fruit was able to ameliorate the effect of flusilazole hepatotoxicity
which may be due to its antioxidant effect and this is evidenced by the
improved hepatic antioxidant enzymes, reduced glutathione and lipid
profile. Further studies are needed to better understand the toxicity
mechanisms of fungicides. In addition focus the ameliorate effects of
date against the deleterious effects of pesticides used.