Dr/ Sherein Saeid AbdElgayed, ph.d

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1 هللامسب

2 Dr/ Sherein Saeid AbdElgayed, ph.d Professor of Veterinary Pathology, Cairo University, Giza, Egypt. Chairman of the Editorial Board of Arab Journal of Science & Research Publishing (AJSRP)

3 Pathological and Immunohistochemical study of Potassium Bromate on the liver of Albino Rats Elmahdy M.M*, Moussa M.H.G**., Sherein, S. AbdElgaied*. And Alwazaan A.A***

4 INTRODUCTION Potassium bromate (KBrO3), is a white crystalline solid and a widely reactive food additive (WHO,1996),

5 INTRODUCTION it is often used in bakeries as flour improver yielding higher bread volume (Kurokawa et al., 1990) and used as a dough conditioner for flour

6 INTRODUCTION Several researches on safety evaluation of potassium bromate were carried out and also It was found to be a genotoxic and carcinogenic.

7 INTRODUCTION Studies have also shown that it possess the potential of inducing cancer, kidney failure, deafness, redness and pains of the eye and skin (Mack, 1988; De Angelo et al., 1998).

8 INTRODUCTION It is classified as a category 2B carcinogen (possibly carcinogenic to humans) based on sufficient evidence of kidney carcinogenicity in rats by the International Agency for Research on Cancer (IARC).

9 INTRODUCTION dose-response relationship in rat kidneys was observed in progressive severity from renal dysplastic foci, preneoplastic lesions, through renal adenomas, and finally renal carcinoma

10 Aim Of The Study The present study was conducted to investigate the toxic and carcinogenic effects of various levels of dietary potassium bromate on the liver of male and female albino rats.

11 Aim Of The Study The criteria for assessment includes; Clinical signs, Biochemical, Histopathological, immunohistochemical

13 Materials & Methods

14 Chemicals Potassium bromate (KBrO3) in the form of powder with a purity of greater than 99.5%. It was supplied by a private chemical company at Cairo, Egypt.

15 Animals A total number of 105 albino rats of both sexes, weighing 120 g were supplied by faculty of Veterinary medicine, Cairo University.

16 Animals They were kept under standard conditions and were housed in metabolic cages and had free access to water and standard diet. The animals were left for a week, as an adaptation period

Experimental Design 105 Albino Rats of both sexes,

female control rats 15 rats given 200 ppm KBrO3 In

KBrO3 In drinking water for 14 months 15 rats

months After 6, 9 &14, 5 animals from male and

17 Experimental Design 105 Albino Rats of both sexes, weighing 120 g 45 Male 45 Female 15 Male and female control rats 15 rats given 200 ppm KBrO3 In drinking water for 14 months 15 rats given 400 ppm KBrO3 In drinking water for 14 months 15 rats given 600 ppm KBrO3 In drinking water for 14 months After 6, 9 &14, 5 animals from male and female Gp And 5 animals From control Gp Were Euthanized For analysis

18 (1)-Biochemical Examination Serum samples were collected and analyzed to determine the level of Aspartate aminotransferase (AST)), and Alanine Aminotransferase (ALT) using the method of Reitman and Frankel (1957)

19 (2)- Histopathological examination Tissue samples of the liver were fixed in neutral buffered formalin 10%, dehydrated in ascending grade of alcohol, cleared and embedded in paraffin, sectioned at 5 μm thickness and stained by H & E and examined microscopically (Bancroft et al. (1996).

20 (3)- Immunohistochemical Examination Proliferating cell nuclear antigen (PCNA) was applied according to (Hall et al., 1990)

22 (1) Clinical Signs

23 (1)- Clinical Signs included; dullness, ataxia and loose their appetite, sometimes circling with paddling movements and hyper-excitability were also recorded in some animals.

24 (2) Biochemical results

25 (2)- Biochemical Results ALT Result

26 Biochemical Results AST

27 (3) Histopathological results

28 (3)- Histopathological Results Fig. (1) H &E, Stained sections of MALE (liver) administered with KBrO3 (H&E X400)

29 Figure (1) A) Male rat liver 6 months (200 ppm), showing congested and dilated blood vessel and necrosis of its surrounding hepatic cells with areas of hemorrhages

30 Figure (1) B) Male rat liver 6 months (400 & 600 ppm), showing increase number of binucleated hepatocytes with variation of nuclear size.

Figure (1) C) Male rat liver for 9 months, showing dilated and hyalinized blood vessel wall with hyperplasia of the bile ducts and formation of newly formed

31 Figure (1) C) Male rat liver for 9 months, showing dilated and hyalinized blood vessel wall with hyperplasia of the bile ducts and formation of newly formed bile ducts and ductules in the portal tract that extended between the hepatic cells which appeared atrophied and deeply stained. Note oval cell proliferation.

32 Figure (1) D) Male rat liver administered for 14 months (600 ppm), showing hyperplasia of the bile ducts with the formation of newly formed bile ducts which arranged in an adenoid pattern

33 Histopathological Results Fig. (2) H &E, Stained sections of FEMALE (liver) administered with KBrO3 (H&E X400)

34 Figure (2) A) Female rat liver administered for 9 months (600 ppm), showing large dysplastic foci of basophilic hepatic cells surrounded by eosinophilic cells.

35 Figure (2) B) Female rat liver administered for 14 months (200ppm), showing trabecular arrangement of small eosinophilic cells and vacuolated cells with hyperchromatic nuclei.

36 (Arrow head). Figure (2) C) Female rat liver administered for 14 months (400 ppm), showing dysplastic appearance of the hepatocytes with hyperchromatic nuclei and clear mitotic cells (arrow) and giant cells

37 Figure (2) D) Female rat liver administered for 14 months (600 ppm), showing dysplastic changes manifested by variations of nuclear size, binucleation and hyperchromatic nuclei with clear large nucleoli.

38 (4) Immunohistochemical results

39 PCNA immuno-reactivity A) Control rat liver after 9 months showing negative (PCNA).

40 PCNA immuno-reactivity B) Rat liver after 6 months showing moderate (200 ppm) intensity of immunostaining of the nuclei and nucleoli of dysplastic cells. PCNA.

41 PCNA immuno-reactivity C) Rat liver after 6 months (400 ppm) of showing strong intensity of immunostaining (PCNA) positive nuclei and nucleoli.

42 PCNA immuno-reactivity D) Rat liver after 9 months showing strong (400 ppm) intensity of immunostaining (PCNA). Note the deeply positive nuclei and light cytoplasmic

43 PCNA immuno-reactivity E) Rat liver after 9 months showing strong (600 ppm) intensity of immunostaining (PCNA) positive nuclei. Note the light cytoplasmic staining

44 PCNA immuno-reactivity F) Rat liver after 14 months showing strong intensity of immunostaining (PCNA) positive nuclei. Note the light cytoplasmic staining

46 Conclusion The present study indicated that a long-term exposure To KBrO3 caused alterations in the histology of the liver in rats.

47 Conclusion KBRO3 cause several degenerative, destructive, necrotic and proliferative changes

48 Conclusion In the present study carcinogenesis was recorded as basophilic, eosinophilic and vacuolated foci of hepatocytes, together with dysplastic changes of hepatocytes.

49 Conclusion The toxic and carcinogenic effects of KBrO3 was dose and time dependent.

51 Recommendation Avoid the hazard use of Potassium Bromate KBRO3

(ICOALS) May

(ICOALS) May هللامسب (ICOALS) 2018 07-09 May Dr/ Sherein Saeid AbdElgayed, ph.d Full Professor of Veterinary Pathology, Cairo University, Giza, Egypt. Chairman of the Editorial Board of Arab Journal of Science & Research