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MANAGEMENT of ACUTE ALUMINUM PHOSPHIDE TOXICITY in RAT MODEL with a NOVEL INTERVENTION, a TRIAL of BORIC ACID *Ola Abdel Hady Sweilum, *Fatma Shaban Kandeel

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MANAGEMENT of ACUTE ALUMINUM PHOSPHIDE TOXICITY in RAT MODEL with a NOVEL INTERVENTION, a TRIAL of BORIC ACID *Ola Abdel Hady Sweilum, *Fatma Shaban Kandeel Sweilum et al. 57 MANAGEMENT OF ACUTE ALUMINUM PHOSPHIDE TOXICITY IN RAT MODEL WITH A NOVEL INTERVENTION, A TRIAL OF BORIC ACID *Ola Abdel Hady Sweilum, *Fatma Shaban Kandeel. **Dalia Abdel Razik Noya *Forensic Medicine and Clinical Toxicology Department, Faculty of Medicine - Menoufia University. **Histology Department, Faculty of Medicine - Menoufia University ABSTRACT Aluminum phosphide (AlP) or rice tablet is a cheap pesticide. When it comes in contact with acid (gastric acid) or moisture, it releases phosphine (PH3) gas. The heart, lungs, liver are the main targets in acute Aluminum phosphide (AlP) poisoning. Most deaths occur due to cardiovascular toxicity. Recently, boric acid has been proposed theoretically as an antidote for AlP poisoning. With the formula B (OH)3, boric acid can accept electrons as it is a Lewis acid with an empty p orbital. The aim of the present study was to investigate the possible protective role of boric acid in management of aluminum phosphide poisoning and its possible use as an effective antidote to reduce ALP poisoning mortality. Forty albino rats, all of which were adults and males were equally divided into four groups, group 1: control (given distilled water), group 2: given boric acid only (100mg/kg nontoxic dose), group 3: given aluminum phosphide (12.5 mg/kg), group 4: given aluminum phosphide followed by boric acid. Rats in group 1 and 2 was killed by cervical dislocation, while rats in group 3 and 4 were left to die, then all were dissected out. Fixation in situ was performed by perfusion technique then the chest and the abdomen were carefully incised to reveal the heart and the liver. The liver and the heart were washed with saline then quickly dissected, excised then blotted for histological preparation. Blood samples were collected from each rat (immediately after death) for measuring liver enzymes (SGOT, SGPT, ALK (alkaline phosphatase) and cardiac troponin. Rats treated with ALP followed by boric acid showed significant increase in their survival time, marked improvement in histopathological changes and also showed significant improvement in biochemical parameters including liver and cardiac markers (SGOT, SGPT, and cardiac troponin) compared with rats intoxicated with AlP. Conclusion: This study may increase the hope of using boric acid as a new effective antidote for ALP poisoning and opens the door of opportunity for larger experimental and then clinical trials to reevaluate the role of boric acid in acute ALP poisoning. Key words: Aluminum phosphide, Boric acid, Toxicity, Cardiac troponin, Antidote. INTRODUCTION (PH3); a colorless, inflammable, water Aluminum phosphide (AlP) is a solid insoluble, and highly toxic gas cheap fumigant pesticide (rice tablet) (Moghadamnia 2012; Bumbrah et al., which protects stored grains from 2012). While odorless in its pure form, insects and rodents with a fatal dose PH3 can smell of garlic or decaying fish between 0.15 and 0.5 grams (0.0053 due to the presence of impurities such and 0.0176 oz) (Wahab et al., 2008). as substituted phosphines and When it comes into contact with diphosphines (Wahab et al., 2009). moisture or acid, it releases phosphine Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 58 Phosphine gas (PH3) is a Lewis humans in nutritional amounts, and is base and a strong nucleophile. It is a thus found in animal and human tissues reducing agent with a lone-pair electron at low concentrations (Environmental which reduces cytochrome c oxidase Protection Agency 2004). It is and interferes with the electron transfer considered as a weak acid and it is non- from complex III to complex IV of the toxic in rats with a median lethal oral mitochondrial respiratory chain, dose of 5.14 g kg-1 (Perelygin and ultimately resulting in the inhibition of Chistyakov 2006). oxidative phosphorylation, adenosine In the study done by Soltani et al., triphosphate depletion and cell death (2016), at Sultan Qaboos University on (Soltani et al., 2013). chemical reaction between boric acid Aluminum phosphide is usually and phosphine, they concluded that taken in suicidal attempt. It was boric acid significantly reduced the rate reported that AlP has a mortality rate of phosphine gas evolution (P <0.01) more than 50% of intoxicated cases and that boric acid may be an efficient (Moghadamnia 2012). After ingestion and non-toxic antidote for PH3 of aluminum phosphide, phosphine gas poisoning. is released in the stomach and after absorption; symptoms and signs of AIM OF THE STUDY poisoning occur rapidly (Wahab et al., This study aimed to investigate the 2008) which include nausea, vomiting, possible antidotal role of boric acid in garlic breath smell, retrosternal and management of Aluminum phosphide epigastric pain, anxiety, agitation, and toxicity and its protective role on Alp- dyspnea (Mostafazadeh 2012 & Popp induced biochemical and histo- et al 2002). pathological changes in liver and heart The heart, lungs and liver are the of adult male albino rats. main target organs in acute AlP poisoning, although multi-organ system MATERIALS & METHODS failure may also occur. Most deaths This experiment was carried out in occur within the first 12-24 hours, the animal laboratory in Faculty of usually because of cardiovascular Medicine, Menoufia University. toxicity (Perry 2007). Animals: Forty adult male albino Patients are usually managed rats weighed 150-240 grams were used conservatively, and their outcome is in this experiment. Animals were very bad as there is no efficient antidote purchased from a small animal breeding has been discovered for AlP poisoning center in Shebin El-kom, Menoufia, till now (Moghadamnia 2012). Egypt. All ethical protocols for animal Recently, boric acid has been treatment were followed. This proposed theoretically as an antidote for experimental protocol was accepted by AlP poisoning. With the formula B the Ethical Committee of Menoufia (OH)3, boric acid is a Lewis acid which faculty of medicine. can accept electrons as it has an empty Materials: Aluminum phosphide p orbital (Soltani et al., 2013). Boric used in experiment was in the form of 3 acid is a colorless, odorless, transparent gm tablets (57% concentration, Excel material used in industrial sector for crop care Ltd, India), and contains two preparation of disinfectants and drugs. compounds: aluminium phosphide and However, it is primarily beneficial for Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 59 aluminium carbonate in a ratio of the abdomen were carefully incised to 57:43. reveal the heart and the liver. The heart Aluminum phosphide Purchased and the liver were washed with saline from local pesticide shop in Shebin El- then quickly dissected, excised then Kom. blotted. Boric acid was obtained from I- Histological study: Tissue toxicology laboratory in toxicology samples were fixed in 10% formol department (boric acid powder saline for 2-5 days then washed in tap dissolved in hot distilled water and water, dehydrated in ascending grades given in a dose of 100 mg/kg dose), of alcohol (50%, 70%, 90% over night Menoufia Faculty of medicine. and 100% for 3 hours), cleared in Substances of experiment were given xylene and embedded in paraffin wax. by gavage (oral tube). Paraffin sections of 5-6 micrometer Experimental design: thickness were cut and stained with Rats were housed in polypropylene hematoxylin & eosin (H &E). cages. Food and water were provided II- Immunohistochemical study: ad libitum (on desired). Rats were given Caspase-3: sections were stained with two days’ time to get acclimatized with the primary rabbit polyclonal anti the laboratory conditions prior to caspase-3 antibody (Thermo Scientific, experiment. All rats were fasted for 24 Lab Vision, USA) (Sanii et al., 2012). hours just before starting the Immunostaining by caspas3 show if experiment (in order to empty gastro there is apoptosis or not, as we want to intestinal tract and abolishing know if ALP produce effect at the level interactions of food components with of the nucleus or not. the tested substance) (vermeulen et al., Biochemical analysis: 1997). Blood samples were collected from Rats were divided into four groups, each rat (immediately after death) for each containing ten rats (n=10). measuring liver enzymes (SGOT, Group 1: Control (given distilled SGPT, ALK (alkaline phosphatase) and water) cardiac troponin. Blood was centrifuged Group 2: Boric acid only (+ve at 3000 rpm to obtain the plasma, then control) in a dose, (100 mg/kg nontoxic SGOT, SGPT activity were determined dose), (Ince et al., 2010). (Ritman and Frankel 1957). Activity Group 3: Given aluminum of alkaline phosphatase was determined phosphide (12.5 mg/kg) (Baeeri et al., (Babson and Read 1959), and the 2013). levels of cardiac troponin were also Group 4: Given aluminum measured by enzyme linked phosphide (12.5 mg/kg) followed by immunoassay (ELISA) technique using boric acid (100mg/kg). a standard kit (Glory Science Co., Ltd, Rats in group 1 and 2 was killed by USA). cervical dislocation, while rats in group Statistical analysis: 3 and 4 were left to die, then all were The collected data were organized, dissected out. tabulated and analyzed using SPSS Histological specimen version 16 software (Spss Inc, Chicago, preparation: ILL Company). The t-test was used to Fixation in situ was performed by compare between mean ± SD of two perfusion technique then the chest and groups. Also, normally quantitative Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 60 data expressed in mean ± SD and was Sections of the liver of rats compared using F test (ANOVA), and intoxicated with Aluminium phosphide the Sig. between groups was done using (group 3) showed destructive changes Post Hoc Test (Tukey). The abnormally in the form of hydropic degeneration of distributed quantitative data was hepatocytes, presence of many expressed in median (Min.
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