Al-Hazmi, N.A.*; El-Sayed Fahim** and Al-Hazmi, M.A.***

* King Abdul-Aziz Univ., Teachers collage** Cairo Univ.,Faculty of Science , Zoology Dept.

*** King Abdul-Aziz Univ., Faculty of Science, Biology Dept.

Neurophysiological and behavioral effects of mycotoxin (patulin)

in albino mice

INTRODUCTION Patulin (4-hydroxy-4H-furo[3,2-c]pyran-

2(6H)-one) is a toxic secondary metabolite produced by a wide range of fungal species of the Penicillium, Aspergillus, Byssochlamys, Eupenicillium and Paecilomycesgenera of which Penicillium expansum, a common contaminant of damaged fruits is the most important (Fuchs et al., 2008).

Penicillium expansum, commonly identified as the ‘‘blue mold rot”, causes the most common storage rot of apples and is known to invade not only fruit but also other foodstuff such as vegetable , berries, bread and meat products (Rychlik & Schieberle, 2001; Ritieni, 2003). For humans, the major potential dietary sources of patulin (PAT) are apples and apple juices made from affected fruit (Murillo-Arbizu et al., 2009).

MATERIALS AND METHODS Animals used in the study were the male albino MF1 mice brought from the center of King Fahad for research in King Abdul-Aziz university, their ages ranged from 10-12 weeks. To study the neurotransmitters, animals were divided into 2 group contained 30 male albino mice and 6 animals were decapitated each week. Group 1 : Control group animals were daily oral administered with saline (0.9 % NaCl) for 6 weeks. Group 2 : animals were daily oral administered with contaminated apple juice with patulin (152.5 µg/ 1ml) for 6 weeks.

To study the behaviors of the animals, the

animals were divided into 2 groups each group contain 10 animals. Group 1: To study Tube Restraint test for the aggressive behavior according to (Al-Hazmi and Brain 1991; Al-Hazmi, 2001) and the following parameters were calculated a- Proportion of animals biting the target . b- The latency period. And c- Number of bites. Group 2: To study locomotors activity test. This test aims to evaluate the difference in the behavioral activity of the animals according to the methods of Al-Hazmi and Brain,1991 ; Al-Hazmi et al., 1997, and the following parameters were calculated : A- Latency period. B- Time of locomotors. C- Time of immobility. D- Number of rear. E- Number of wall rear and F- Number of squares crossed.

Neurotransmitters measurement: Dopamine (DA), norepinephrine (NE) and serotonin (5-HT) were estimated and determined according to the method of Chang, 1964 modified by Ciarlone 1978, by spectroflorometric method (Tyrner quantech Tm, Digital filter, Branstead Thermoline, Corporation, Asuvsidlary of Sybaon, International Company, U.S.A.). Brain areas were dissected according to the method of Glowinski and Iversen, 1966, into 7 regions which are : Cerebellum, Pons plus medulla oblongata, Striatum, Cortex, Hypothalamus, Midbrain and Hippocampus.

Table (1): Effect of chronic oral administration of apple juice contaminated with patulin (152.5 µg/1ml) on dopamine (DA) content in the different brain areas of albino mice.

CerebellumPons & MedullaStriatumCortexHypothalamusMidbrainHippocampus

2 weeks

Control384 ± 1.5508 ± 3.5815 ± 4.7216 ± 1.3655 ± 3.7557 ± 402981 ± 7.1

Mean ± S.E341 ± 2.9630 ± 2.91119 ± 6.3296 ± 2.5961 ± 4.9575 ± 6.31107 ± 9.4

%change-11.20%24.02%37.30%37.04%46.72%3.23%12.84%

significant************n.s*

4 weeks

Control433 ± 1.9532 ± 3.1824 ± 3.1238 ± 2.0612 ± 4.0500 ± 3.1838 ± 6.3

Mean ± S.E412 ± 3.3660 ± 5.0924 ± 503267 ± 1.9834 ± 906526 ± 4.7987 ± 6.6

%change-4.95%24.06%12.1412.18%36.27%5.217.78%

significantn.s*******n.s*

6 weeks

Control314 ± 3.2565 ± 2.7634 ± 2.7187 ± 1.2608 ± 5.1403 ± 2.7667 ± 4.5

Mean ± S.E320 ± 5.6664 ± 3.2822 ± 3.5237 ± 1.5846 ± 7.0397 ± 2.1863 ± 5.6

%change+1.88%17.5229.6526.74%39.14%-1.49%29.39%significantn.s**********n.s***

n.s. non-significant change according to Student,s T-test. *significant change ( P< 0.05 ) according to Student ,s T-test.

**highly significant change ( p< 0.01 ) according to Student ,s T-test. ***more highly significant change ( p<0.001 ) according to Student ,s T-test.

Table (2): Effect of oral administration of apple juice contaminated with patulin (152.5 µg/ 1ml) on norepinephrine (NE) content in the different brain areas of albino mice.

 

CerebellumPons & Medulla

StriatumCortexHypothalamusMidbrainHippocampus

2 weeks

Control466 ± 3.1545 ± 2.7852 ± 3.9239 ± 1.6853 ± 5.7586 ± 3.71004 ± 6.6

Mean ± S.E

446 ± 2.9523 ± 1.91047 ± 5.1295 ± 1.7968 ± 7.1519 ± 2.91045 ± 7.2

%change-4.29%-4.04%22.89%23.43%13.48%-11.43%4.08%

significantn.sn.s******n.s

4 weeks

Control436 ± 2.1567 ± 3.9592 ± 3.6219 ± 1.5623 ± 4.1512 ± 2.1905 ± 4.6

Mean ± S.E

463 ± 2.7566 ± 5.6867 ± 4.1260 ± 1.9773 ± 4.2525 ± 3.4937 ± 5.2

%change6.19%-0.18%46.45%18.72%24.08%2.54%3.20%

significantn.sn.s******n.sn.s

6 weeks

Control317 ± 2.8575 ± 3.1669 ± 4.7180 ± 0.9580 ± 2.9423 ± 2.3741 ± 3.4

Mean ± S.E

324 ± 3.5597 ± 4.2862 ± 5.3203 ± 1.3723 ± 3.7438 ± 3.0820 ± 5.0

%change2.21%3.83%28.85%12.78%24.66%3.55%10.66%

significantn.sn.s******n.sn.s

n.s. non-significant change according to Student ,s T-test. *significant change ( P< 0.05 ) according to Student ,s T-test.

**highly significant change ( p< 0.01 ) according to Student ,s T-test. ***more highly significant change ( p<0.001 ) according to Student ,s T-test .

.

Table (3): Effect of oral administration of apple juice contaminated with le patulin (152.5 µg/ 1ml) on serotonin (5-HT) content in the different brain areas of albino mice.

CerebellumPons & Medulla

StriatumCortexHypothalamusMidbrainHippocampus

2 weeks

Control421±3.0678 ± 3.7738 ± 4.9210 ± 1.0707 ± 3.0560 ± 3.6965 ± 5.0

Mean ± S.E 428±1.6±692 ± 4.0895 ± 3.7252 ± 1.7918 ± 5.4574 ± 3.11009 ± 7.1

%change1.66%2.06%21.27%20.0%29.84%2.5%4.56%

significantn.sn.s******n.sn.s

4 weeks

Control425±3.2554 ± 3.1790 ± 3.1231± 1.9619 ± 3.3466 ± 2.9806 ± 6.7

Mean ± S.E410 ± 2.9630 ± 4.2853 ± 4.9292 ± 0.7737 ± 4.3530 ± 1.7971 ± 5.1

%change-3.53%13.72%7.97%+26.41%19.06%13.73%20.47%

significantn.s*n.s******

6 weeks

Control345 ± 2.6592 ± 9.3656 ± 3.7188 ± 1.3622 ± 5.1418 ± 2.1696 ± 3.9

Mean ± S.E360 ± 3.1673 ±3.6797 ± 4.0281 ± 1.5768 ± 4.9521 ± 3.0789 ± 4.1

%change4.35%13.68%21.49%49.47%23.47%24.64%13.36%

significantn.s***********

n.s. non-significant change according to Student,s T-test. *significant change ( P< 0.05 ) according to Student ,s T-test.

**highly significant change ( p< 0.01 ) according to Student ,s T-test. ***more highly significant change ( p<0.001 ) according to Student ,s T-test .

Table (4): Explain (Range ± Median) on the effect of apple juice contaminated with patulin (152.5 µg/1ml) on the locmotors behavior of albino mice.

BehaviorDuration of (in sec.) number of

Drµgs doses

)152.5 µg/1ml(

Latency to

explore first hole

+

Locomotion

+++

Immobility+++

Rears+++

Wall rears+++

Squares crossed

+++

0.0Control

3)2-3(

551.5)546-557(

45.50)40-51(

56.5)43-74(

152)134-171(

751)734-762(

Patulin3

)2-3(542.5*

)529-567(55*

)30-67(19.5***

)5-41(76.5***

)49-112(659***

)642-753(

+significant ( p< 0.05 ) according to Kruskall Wallis test +++more highly significant ( p< 0.001 ) according to Kruskall Wallis test.

*significant change ( p< 0.05 ) according to Mann-Whitney U test. ***more highly significant change ( p<0.001) according to Mann-Whitney U test .

Doses)152.5 µg/1ml(

bite Latency (second) to First)Median with Range(

+++

Number of bites)Median with Range(

+++0.0

control15

)10-25(192.50

)177-233(

patulin25**)15-40(

107***)55-198(

Table (5): Explain (Range ± Median) on the effect of apple juice contaminated with patulin (152.5 µg/1ml) on the aggressive behavior of albino mice.

+++more highly significant ( p< 0.001 ) according to Kruskall Wallis test. **highly significant change ( p< 0.01 ) according to Mann-Whitney U test.

***more highly significant change ( p<0.001) according to Mann-Whitney U test

Thank You