Bioremidation of Spirulina Plantesis Against Deltamethrin Mediated Toxicity and Its Chemical Residues in Chicken Meat

Deltamethrin (DM) is a synthetic pyrethroid insecticide commonly used in veterinary and agriculture. However, both animal and human exposure is associated with hepatorenal toxicity. Our experimental goal was to assess the protective effects of Spirulina plantesis (SP) against DM-induced hepato-renal injury, growth performance, meat chemical composition and its residues in meat, liver and skin of broiler chickens. Sixty one-day-old Cobb broiler chicks were assigned to 4 experimental groups, each was divided into 3 replicates with ve chicks each; 1 st group received basal diet alone (Control), 2 nd group supplemented with SP 20 g/kg diet, 3 rd group supplemented with DM 300 mg/kg diet, and 4 th group supplemented with DM and SP diet. All groups were received the treatment for 35 days. DM, decreased body weight, weight gain, and increased feed conversion rate. DM showed signicant increase in ALT, AST, urea, creatinine and MDA and signicant decrease in SOD, and GSH levels. Also, signicant decrease in DM levels in meat, skin, and liver by SP with a reduction % 63.01, 63.00 and 62.90 % in meat, skin and liver, respectively. The DM intoxicated group showed signicant decrease in protein and signicant increase in fat, cholesterol and triglycerides when compared to control group. Histopathological changes were also recorded. Dietary SP improved these parameters parameters. Dietary inclusion of SP can be recommended due to the protective effects on DM induced toxicity in broiler chickens.


Introduction
By 2050, the demand for animal protein will increase. Chicken meat has been identi ed as a major Therefore, the main aim of this study was to evaluate the bioremediation role of SP as a feed supplement for broiler chicken against DM effect on body weight gain, feed conversion rate, hepatorenal toxicity, meat chemical composition and its residues in meat, liver and skin.

Materials And Methods
Chemicals DM (Butox ® 50 mg/ml; Intervet Co., France) for veterinary use, while SP was purchased as a powder from HerbaForce Co., UK.

Experimental broiler chicks
Sixty, one-day-old Cobb broiler chicks (El-Wataniya Poultry Company, Egypt). Chicks were assigned to 4 experimental groups, each was divided into 3 replicates with ve chicks each; 1 st group received basal diet alone (Control), 2 nd group supplemented with SP 20 g/kg diet (Mirzaie et al. 2018

Growth parameters:
Daily feed intake and gain in body weight were calculated. FCR was calculated after the end of the experiment (total feed consumed divided by a total weight gain).

Blood samples
Blood samples were collected via wing vein from all chickens in each group after the end of the experiment (35 days), in clean, dry tubes and it was left in a slope position to clot at the room temperature. After blood centrifugation at 2000 g for 10 min, serum samples were obtained and it was kept frozen at-20°C until further use in biochemical analysis.

Detection of oxidative cascade indices:
The tissues (liver, kidney) was dissected and washed with a phosphate buffered saline solution (pH 7.4) containing heparin to remove any clots or red blood cells. One gram of each tissue was homogenized in buffer (5 ml), using homogenizer. Tissue homogenates was centrifuged at 4000 rpm for 20 min, then stored at -20 ºC. Oxidative status was done by determination of Glutathione (GSH) level, malondialdehyde (MDA) level and superoxide dismutase (SOD) were evaluated using commercial kits (Biodiagnostic Co., Egypt).
Evaluation of meat chemical composition: Protein and fat content of chicken's meat samples were estimated as described by Anderson (2007). Cholesterol and triglycerides were evaluated spectrophotometrically at wavelength 578nm (Rifal et al. 1999).

Histopathological examination:
Samples from liver, kidney and intestine were collected from slaughtered chickens and prepared according to Banchroft et al. (1996).

Statistical Analysis:
The results (mean±SE) were analyzed using one-way ANOVA, then to determine the differences between the averages; Duncan's test was applied. Statistical Package for Social Science software (SPSS (20) software (SPSS Inc., Chicago, USA) was used.

Results
Effect on on growth performance parameters: Broiler chickens exposed to DM, revealed signi cant reduction in BW, BWG, and an increase in FCR. SP treated group showed an increase in BW, BWG, and decrease in FCR. DM+SP treated group showed an improvement in growth performance compared to control group (Table 2).
Effect on liver markers (ALT, AST) and kidney markers (Urea, Creatnine): Serum of broiler chicken exposed to DM showed signi cant increase in ALT, AST, urea and creatinine. Non-signi cant changes were recorded in SP treated group in relation to control group. While serum of broiler chicken treated with DM + SP group showed signi cant decrease in ALT, AST, urea, and creatinine when compared to DM treated group (Table 3).
Effect on oxidative cascade indices in liver and kidney tissues: Homogenate of liver and kidney tissues of DM treated group showed a signi cant increase in MDA and signi cant decrease in SOD, and GSH levels. Also, SP treated group showed non-signi cant change when compared to control group. While, DM + SP treated group showed signi cant decrease in MDA and signi cant increase in SOD, and GSH levels when compared to DM treated (Table 4).
HPLC evaluation of DM residues in meat, liver and skin of broiler chickens: DM failed to be detected in non-intoxicated groups (control and those fed SP alone). There was a signi cant difference between groups intoxicated with DM and those treated with DM+SP. Results in table (5) and gure (1) revealed a signi cant decrease in DM levels in meat, skin, and liver by SP with a reduction percentage 63.01, 63.00 and 62.90 % in meat, skin and liver, respectively.
Effect on Meat chemical composition (protein, Fat, Cholesterol and Triglycerides): SP in poultry diet has a direct effect on protein, fat, cholesterol and triglycerides in meat. It also decreases DM toxic effect on meat as shown in Table (

Discussion
Since DM is widely used as an insecticide, its effect on the human health and the environment has become a signi cant concern (Sibiya et al. 2019). DM induced histological changes in various organs. DM was metabolized in the liver; toxic DM metabolites have been found in various tissues, including, liver, brain, fat and muscles (Abdelkhalek et al. 2015).
In the current experiment, DM supplementation to broiler chicken, elicted toxicological effects in different tissues; which was reported through biochemical, residual effect on chicken meat, liver and skin and its effect on chicken meat quality and the hisopathological investigations. SP supplementation provided a bioremediation role.
Regarding to the effect of DM on the body weight our result revealed that there are signi cant decreases in the body weight of DM treated group compared to control and SP treated groups. This study mentioned that an increase in growth performance in SP treated group. Previous studies have indicated that SP supplementation has a positive impact on poultry growth ( SP is a type of blue, green algae that contains all essential AAs as well as the pigments phycocyanin (strengthens immunity) and chlorophyll (helps with detoxi cation) (El-Tantawy 2015). SP is well-known for its high nutritional value and a strong hepatic and renal protective activity; it also has an important function in many toxicitiy studies (Mazokopakis et al. 2014). SP administration decreased serum hepatic and renal injury biomarkers and lowered lipid peroxidation in tissues. SP's antioxidant and protective properties were due to the presence of antioxidant active constituents such as minerals, vitamins, proteins, carbohydrates, lipids, b-carotene, and C-phycocyanins, (Upasani and Balaraman 2003).
Nowadays, Food safety becomes a major concern. Chemical contamination of meat with pesticide residues causes adverse impacts on human health. In this study, meat, skin, and liver samples were evaluated for DM residues. As, pyrethroids are lipophilic in nature and precipitate into fatty tissues, therefore, the main target organ is fat for surveillance purposes (Stefanelli et al. 2009). It was found that the higher DM residual levels were in skin followed by liver and meat. This may be due to that skin of poultry (rich in fat), liver also tend to form fat deposition  (Bhilwadikar et al. 2019). So, there is a crucial need for evaluate effective, sustainable, and environment-friendly pesticide removal practices (Azam et al. 2020). In this study, SP lowered DM residues with reduction percentages 63.01, 63.00 and 62.90 % in meat, skin and liver, respectively. This agrees with Abdel-Daim et al. (2016) who normalize toxic effects of DM in mice by SP. In recent decades, Poultry meat consumption has increased. As, it is an important source of protein of growing concern worldwide (Daghir et al. 2020).
Concerning the effect of DM and\or SP on meat chemical composition. It was found that the meat chemical composition in the group fed with SP displayed a large rise in protein while decreasing in fat, cholesterol, and triglycerides when compared to control group. While, meat samples in the DM-intoxicated group had a substantial decrease in protein but an increase in fat, cholesterol, and TG when compared to control group. Moreover, it was found that meat samples from a DM+SP treated group revealed a signi cant increase in protein and signi cant decrease in fat, cholesterol and TG. Results revealed that intoxication with DM caused an increase in fat, cholesterol and TG. A decrease in protein levels in intoxicated groups has been detected. The higher fat content in tissues, the greater amount of pyrethroids. There is a strong association between lipid content and pyrethroid levels in beef and sh, while chicken samples show a mean correlation, may be due to chickens are slaughtered at an earlier age The histological ndings (liver, kidneys, and intestine) in SP treated group were similar to those of the control group revealed that SP did not cause any adverse effects on the chicken tissues examined. DM in chickens showed disrupted hepatic, renal and intestinal architecture. Thus, changes in histopathology could be caused by the production of ROS, which causes damage to various tissues (Abdel-Daim et al.

Conclusion
The role of oxidative stress in DM toxicity in broiler chickens is critical. Antioxidants have been shown to be effective in preventing the toxicity caused by DM. DM toxicity resulted in varying degrees of lipid peroxidation, suppression of antioxidant enzymes′ and glutathione function, and changes in serum biochemical parameters in this study. In terms of serum and tissue biochemical changes, antioxidant activity, and oxidative stress, SP supplementation provided nearly complete protection. It causes a marked decrease in DM residues in meat, skin and liver. Moreover, enhances chemical composition of meat.

Declarations Ethics approval
This study was approved by the Ethical Committee of the Faculty of Veterinary Medicine, Benha University, (BUFVTM 06-03-21).

Consent to Participate
All authors interpreted the data and approved the nal version.

Consent for publication
The manuscript is not previously published in the same or very similar form in other