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Effects of phosalone consumption via feeding with or without sodium bentonite on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep



Transfer of pesticides from environment to animal products is inevitable, so the purpose of the present work was to evaluate phosalone consumption via feeding with or without sodium bentonite (SB) on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep.


Twenty Baluchi ewes were divided into four treatments (P1 as control, P2, P3, and P4) of five animals in which phosalone, an organophosphate pesticide, was given via diet (only for P2 and P3) at a dose of 280 mg/sheep/day for 63 consecutive days. The SB (32 g/sheep/day; for P3 and P4) was also evaluated for its ability to reduce deleterious effects of phosalone in the sheep diets. The control group (P1) did not receive any phosalone and SB during the experiment. Sampling was conducted in two periods of time including weeks 5 and 9.


Phosalone residues were observed in the milk samples of P2 and P3 groups during two sampling periods. During period 1, the transfer rate of phosalone from feed to milk was 0.23 and 0.02%, respectively for the contaminated diets (P2 and P3), which is relatively similar to period 2 (0.22 and 0.02%). Only 0.34 (period 1) and 0.36% (period 2) of phosalone residue are excreted in the feces of P2 group following its daily consumption. Transfer of phosalone from feed to milk was affected (P < 0.05) by the dietary inclusion of a commercial SB, as it (SB) decreased excretion of phosalone via milk (P3). The phosalone and SB alone or together had no significant effect (P > 0.05) on the dry matter intake (DMI) and body weight (BW) gain, but feed efficiency, milk production, milk fat, dry matter (DM) and organic matter (OM) digestibility, acetylcholinesterase (AChE) inhibitory activity, hemoglobin (Hb), red blood cell (RBC), serum glutamic pyruvic transaminase (SGPT), serum glutamic oxaloacetic transaminase (SGOT), albumin and mean corpuscular hemoglobin concentration (MCHC) affected by the treatments in period 1 or 2 (P < 0.05). The Hb, RBC, and MCHC were significantly decreased (P < 0.05) by about 9.72, 20.77, and 9.71%, respectively in the group P2 as compared to those of the control group during period 1. The AChE inhibitory activity (period 1 and 2) significantly increased when phosalone administered via the diet (P < 0.05).


Although there were no adverse effects on the performance of sheep following the intake of phosalone alone (P2 vs. P1), but other research on the long and short times to the phosalone in high and low doses with more animals is suggested. Overall, compared to the control group, addition of SB in the diet of sheep improved nutrient digestibility, animal performance, and milk health.


Animal feed is sometimes contaminated with pesticide residues [69] and these residues may be passed into the body and animal products [68]. In some cases, residues of pesticides are detoxified by the biological system of the body and excreted in urine and feces [38, 52]. Pesticides are materials with the chemical or biological origin that used extensively by human society and can be extremely toxic and harmful to both people and animals. The hazard of cancer via pesticide exposure have examined by many researchers [33, 62, 89]. In order to control pests in Iran, about 5261.80 tons pesticides are used in 2013 [29]. Phosalone is an organophosphate pesticide (OP) that has been widely used by Iranian farmers. Phosalone residues in food and feed with different concentration have been reported by researchers in Iran and other countries [76, 88, 93]. Low levels of phosalone were found [80] in fat and liver (only at 100 ppm) of beef cattle and sheep when administrated in feed at concentrations of 10, 30, and 100 ppm. No residues of phosalone were observed in the milk of dairy cattle at levels of 25, 50 or 100 ppm [81]. Organophosphate pesticides (OPs) are considered as inhibitors of AChE activity and subsequently, accumulation of excessive acetylcholine occur in synaptic cleft available in any live organisms [19, 52, 73]. The residues of several pesticides such as phosalone (0.29 ng/g) in human breast milk were detected by Sharma et al. [85]. Toxic effects of phosalone administered in the feed of rats (0, 5, 50, or 1000 ppm) were investigated by Barker and Sortwell [9] for two consecutive years, but the concentration of 1000 ppm being reduced to 500 ppm at week 27. The Clinical effects such as abnormal posture, hypersensitivity, depression of AChE activity, and poor grooming were observed in groups exposed to 500 ppm phosalone. The enlargement and foamy change in the zona glomerulosa of adrenal gland were observed in rats exposed to 500 ppm. Intestinal epithelial proliferation was found in the normal rat intestinal cells treated with diazinon [35]. Reddy et al. [77] reported hypoventilation and a hypoxic condition in rats exposed to sublethal doses of phosalone. Many methods have been employed for the removal of pesticides and their destructive effects on the body of animals (e.g. use of activated carbon and phenobarbital feeding by Cook and Wilson, [20]; application of atropine as an antidote against OPs by Proskocil et al. [75]; α-lipoic acid supplementation by Al-Attar, [4]). Bentonite as volcanic clay is being used with different goals in industrial agriculture today. Supplementation of contaminated diets with SB decreased transition of aflatoxin M1 to goat’s Milk [70]. Also bentonite has been used as bleaching [48], remove pathogens from the gastrointestinal of poultry [74], toxin binder [56], stabilization of sewage sludge containing heavy metals [49], buffering agent in feedstuffs [24], manipulation of a rumen ecosystem [39, 67] and performance of broiler chickens [55]. The performance of fattening Zandi lambs improved with the dietary inclusion of SB compared to control group [54] and also, fat-tail percentage decreased. The SB participates by manipulation of volatile fatty acid (VFA) profiles, decreasing the dilution rate and subsequently slowing passage rate, changing of the ion exchange capacity of minerals and inactivation the harmful health effects of mycotoxins [47]. There was very little information about transferring of phosalone from contaminated diets to milk and its impacts on blood metabolites and performance of sheep. Although the effects of SB inclusion in the diet of livestock have been studied enormously, it is unknown whether the SB administration via feeding affect ability of animals for reduction of phosalone residues in Iranian Baluchi sheep’s milk or not, hence the present study was conducted for evaluating the effects of phosalone consumption via feeding with or without SB on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep.


Animals, diet and treatments

This experiment was conducted at the Research Station of Ferdowsi University of Mashhad located in Mashhad-Ghochan road. Twenty Iranian Baluchi ewes (45 ± 2.5 kg BW, first-parity and early lactation) were randomly allocated to four treatments (P1, P2, P3, and P4) of five ewes each, fed a total mixed ration. Phosalone, an OP, was daily sprayed on the feed and fed to the ewes according to the following method: Treatment P1 = control without phosalone and sodium bentonite (SB); Treatment P2 = phosalone administered in a dose of 280 mg/day/ewe; Treatment P3 = phosalone (280 mg/ewe/day) and SB (32 g/ewe/day) fed for 63 consecutive days; Treatment P4 = feeding SB with concentration of 32 g/ewe/day. In this experiment, the level of 280 mg/day/ewe set based on the existing information on the milk of animals at different levels of consumption. We selected the mentioned dose to ensure enough quantity of phosalone residues would reach the milk to evaluate neutralized effect of SB. The SB with a commercial name of Zarin Binder was supplied by the Vivan Company located in Mashhad, Iran. Phosalone with a pure analytical grade (99.3%) was dissolved in acetone and immediately mixed with a small part of the diet. After evaporation of acetone, contaminated diets offered to the animals. The feed container of animals was checked regularly to insure that no residues of contaminated feed remain. To ease the separation of urine and feces, the ewes kept in individual metabolic cages. Animals were housed in an environment protected from wind and rain. The ewes were fed at 07:30 a.m. and 17:30 p.m. with a total mixed ration (Table 1) containing forage and concentrate in a ratio of 65:35. The diet was formulated according to the recommendation of NRC [72]. Following the 14 day adaptation period, animals were randomly allocated to treatments. The ewes were given free access to fresh water and milked twice a day (06:30 a.m. and 16:30 p.m.).

Table 1 Ingredients and chemical composition of an experimental diet fed to Iranian Baluchi sheep

Animal sampling

Feed intake, ort and milk yield were recorded daily for each animal. Likewise, feed samples were taken weekly for determination of dry matter and chemical composition. Blood samples (weeks 5 and 9) were gathered just 3 h post the morning feeding in heparin tubes via jugular vein (5-10 mL), centrifuged (3000×g, 10 min), and the supernatant containing plasma fluid was drawn into sterile 1.5 mL micro-tubes and then conserved at −80 °C for the further analysis. Also samples of blood without centrifugation were quickly transferred to the laboratory for hematology analysis. Samples of urine and feces for phosalone residues were collected in weeks 5 and 9 (in 5 successive days) following the different feeding treatments. Ewes were also weighted before morning feeding in weeks 0, 5 and 9 for gain increment. During the collection periods (5 consecutive days in weeks 5 and 9), daily samples of feed and feces were regularly collected and then were mixed together (with a specified ratio) and one subsample considered for next analysis. The milk of 5 consecutive days was sampled from ewes in weeks 5 and 9, mixed basis milk yield and a subsample analyzed for chemical composition and phosalone residues.

Laboratory instructions

The dry matter (DM), crude protein (CP), ether extract (EE), Ca and P contents were assessed by the methods of AOAC [6]. The method of Van Soest et al. [96] was applied for determination of neutral detergent fiber (NDF) and acid detergent fiber (ADF). Milk composition, including protein, fat, lactose, and total solid were measured using a milkoscan analyzer (Foss Electric, Conveyor 4000, Hillerød, Denmark). Dry matter and organic matter digestibility were calculated by measuring their concentrations and the values of acid insoluble ash (AIA) as an internal marker in the samples of feed and faecal [95]. The QuEChERS method [7] followed by gas chromatography–mass spectrometry (GC/MS) with selected ion monitoring (SIM) was applied for the determination of phosalone residues in the faecal, urine, and milk samples. Analysis method for GC/MS included: oven temperature of 200 °C; injection temperature of 250 °C; column flow of 1 mL/min; ion source temperature of 200 °C; interface temperature of 300 °C; carrier gas of helium with purity >99.999%; and mode of splitless. Extracted samples of 1 μL were injected into GC/MS apparatus. The analytical capillary column was an Rxi-1 ms (length: 30 m, internal diameter: 0.25 mm, df: 0.25 μm; manufactured by Restek of USA). The blood urea nitrogen (BUN), glucose, cholesterol, albumin, serum glutamic pyruvic transaminase (SGPT), serum glutamic-oxaloacetic transaminase (SGOT), and total protein were determined using an auto-analyzer (Biosystems A15; 08030 Barcelona, Spain). The AChE activity of plasma was measured by the method of Elman et al. [26] and enzyme inhibitory activity of each sample as percentage was calculated from the slope (plot between absorbance and time) of the line with or without inhibitor [65]. Determination of hemoglobin (Hb), white blood cell (WBC), red blood cells (RBC), packed cell value (PCV), mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), and mean corpuscular hemoglobin concentration (MCHC) were performed using an automated hematology analyzer (CellTac α, MEK-6450, Nihon Kohden, Japan).

Statistical data analysis

All data for each period were separately analyzed with a completely randomized design using PROC GLM of SAS software (Version 9.1, SAS Institute). Each animal was considered as experimental unit. Statistical model was: Yij = μ + Ti + εij, where Yij is the dependent variable; μ is the overall mean; Ti is the treatment effects; εij is the residual error. Differences between treatment means were determined by Duncan’s multiple range test at level of P < 0.05.


Feed intake and body weight changes

DMI, Feed efficiency, OM or DM digestibility, and weight characteristics of ewes following treated ration consumption in two experimental periods are given in Table 2. During the two experimental periods, DMI was similar between treatments (P > 0.05), but treatment containing SB in period 2 (P4) had significantly higher (P < 0.05) feed efficiency, DM and OM digestibility compared the control group (P1). During period 1 and 2, the ewes fed SB had a numerically higher BW (P > 0.05). Feeding of phosalone to the ewes (P2) caused no significant effect on OM or DM digestibility compared to control group (P > 0.05).

Table 2 DM intake, Feed efficiency, OM and DM digestibility, and weight characteristics of ewes following treated ration consumption in two experimental periods

Milk composition and its yield

Milk composition and lactation yield of ewes fed the experimental treatments in two sampling periods are presented in Table 3. When SB was added to the diets, milk yield as kg/day or 6% FCM increased (P < 0.05) during period 2 and the highest value was observed for the ewes fed treatment P4. There was no significant effect on milk yield (kg/day or 6% FCM) between treatments P1 and P2 (P > 0.05) during period 2. Supplementation of the diet with SB increased milk fat (P < 0.05), but protein, lactose, and total solids not affected by the treatments (period 1 and 2).

Table 3 Milk composition and lactation yield of ewes fed the experimental treatments in two sampling periods

Hematology and plasma metabolites

Hematology and plasma metabolites of ewes fed with the experimental treatments in two sampling periods are shown in Table 4. Plasma AChE inhibitory activity was significantly higher (P < 0.05) in the ewes fed phosalone alone (P2) compared with control group, but glucose, BUN, cholesterol, and total protein were not affected by the treatments during period 1 and 2. Also during period 1, SGPT and SGOT were significantly decreased following phosalone administration in the diet compared to control group (P < 0.05). The highest albumin was observed in the treatment containing SB. The addition of SB to the diet had no significant effect on Hb, RBC, and MCHC than the control group; but these parameters were decreased following the dietary inclusion of phosalone (P2) during period 1 and 2. The hematological parameters such as WBC, PCV, MCH, and MCV were not influenced by the treatments in this experiment (P > 0.05).

Table 4 Hematology and plasma metabolites of ewes fed with the experimental treatments in two sampling periods

Phosalone residues

Monitoring the phosalone residues in ewes fed by the experimental treatments in two sampling periods are presented in Table 5. There was no phosalone residue in treatments P1 and P4, so they withdrew from the statistical analysis. Residues of phosalone were detected in the samples of milk, faecal and urine of both P2 and P3 treatments during period 1 and 2. The addition of SB in the diet decreased transfer rate of phosalone to milk (P < 0.05).

Table 5 Monitoring the phosalone residues in ewes fed the experimental treatments in two sampling periods


Feed intake and body weight changes

The DMI and body weight of lactating crossbred goats were not affected due to feeding monocrotophos at a dose of 25 mg/kg (DM basis) [57]. In the present work treatment of diet with the phosalone had no effect on BW of ewes compared to the control group during two sampling periods. In studies of Sangha et al. [82], feed intake and water intake were affected by cypermethrin at a dose of 50 mg/kg BW in treated rats [82], also the body weight of rats decreased significantly compared to the control group (P < 0.05). In this study, supplementing the diet of ewes with SB increased OM and DM digestibility compared with the control group (P < 0.05) during period 2. SB is classified as expanded lattice clay in a montmorillonite group of minerals [10]. Having a high potential for ion exchanging in the bentonite can bind a different range of cations [31]. Wool growth improved by supplementation of SB in the sheep diets [17, 30, 31]. Also, SB reduced the concentration of ruminal ammonia and improved the passage of feed and microbial protein to the small intestine [42]. The feed intake and average daily gain of lambs were increased (P < 0.05) following SB supplementation [98]. In our study, the average DMI of periods 1 and 2 was 1.24, 1.33, 1.28 and 1.40 Kg/day for the P1, P2, P3, and P4 treatments, respectively, but the differences between treatments were not significant (Table 2). Similarly, feed intake expressed as kg/kg BW, as well as kg/kg MW was not significant (P > 0.05). In a study of Berthiaume et al. [11], although supplementation of direct-cut grass silage with bentonite increased the BW gain of steers (P < 0.05), it had no effect on DMI and nutrient (DM, OM, NDF, ADF, and nitrogen) digestibility of grass silage. It was found that ciliate protozoa could not consume bentonite interring the rumen; however bentonite interfered with movement efficiency of cilia and subsequently prevented the motility of protozoa, which diminished the power of predation for rumen bacteria [97]. Population of protozoal in ruminal fluid of faunated rams was lower for animals fed bentonite or monensin than to control group [40]. In this experiment the BW gain (kg/day) was comparable between treatments, ranging between 0.018, 0.024, 0.025 and 0.026 kg/day (average period 1 and 2) respectively in P1, P2, P3 and P4 treatments. An increasing trend for DMI and BW gain was observed in the treatments containing SB. The nitrogen utilization was improved due to the feeding of soybean meal with bentonite [16]. The growth performance of lambs was also improved when fed a diet containing 1% of SB [98], but supplementation of a diet with 2.5% decreased the performance of finishing steers [18]. The VFA, BW, DMI, faecal excretion, apparent digestibility of DM, OM, CP, NDF, and ADF were not affected by the 1% SB in the diet of growing lambs compared to the control group [3].

Milk composition and its production

Due to increase of DMI in the diet containing SB, there was a trend to increase undegradable intake protein (UIP) in the lambs [98]. The increased passage of microbial protein and feed to the small intestine occurred when SB was used [41]. In our experiment (period 2), an increase in milk production may be related to feed improvement and microbial protein supply to the small intestine following SB consumption. It is predicted that higher more bypass protein from the rumen to the small intestine will increase ruminant performance, especially for the meat and milk containing more protein composition. Although SB is suggested to change ruminal microbial populations, but there was observed only an increasing in total VFA (P < 0.05) concentrations [98]. In one study conducted with Colling et al. [18], they found an increase in acetate and butyrate concentrations and a decrease in propionate following SB supplementation [18]. In the current research supplementation of the diet with 280 mg of phosalone/day/ewe (P2) had no significant effect on milk yield and milk components (Table 3) compared to the control group (P1), but diet containing SB had a higher milk yield and milk fat during period 2. The application of bentonite in the sorghum grain-based diets did not change the milk composition or production in Holstein–Friesian dairy cows, but the rumen pH increased (P < 0.05), faecal starch and rumen ammonia decreased and animals tended to intake less grain sorghum [25]. Similarly, it has been observed that dietary supplements of bentonite diminished microbial degradability of feed protein and increased microbial protein synthesis in the rumen, the small intestine flow of amino acids, and growth of wool in sheep containing regular ruminal microbial population [41]. Although bentonite in P3 could greatly reduce the adverse effects of phosalone in the animals, part of toxicity effects on animal metabolism following excessive intake of phosalone still seems to remain. So in the present study, SB alone (P4) increased the productivity of ewes (i.e., milk yield and milk fat), but phosalone with SB (P3) did not affect those compared with control group during sampling periods.

Hematology and plasma metabolites

The addition of chlorpyriphos in the diet of calves increased (P < 0.05) glucose, ALT, and AST and decreased (P < 0.05) the AChE activity [84]. The carp fish were exposed to low (0.15 mg/L), medium (0.3 mg/L), and high (0.6 mg/L) levels of phosalone for 14 consecutive days. The RBC, WBC, hematocrit, Hb, MCV, MCH, and MCHC were significantly affected by levels of phosalone [51]. Oral administration of diazinon at a level of 50 mg/kg BW for 21 days did not significantly accompanied with pathological and clinical symptoms following decreasing the AChE activity in the sheep [5]. In this study, no sign of toxicity was observed until the end of the experiment. The Activity of AChE in the erythrocytes, plasma or serum of sheep after dipping in the diazinon soluble was not affected by pesticide [37]. The AChE is an enzyme which breaks down the acetylcholine into acetate and choline; hence OP can block the activity of this enzyme [36]. The OP compounds inhibit AChE which hydrolyses acetylcholine. Binding of OP with AChE caused to phosphorylation of the enzyme and this reaction is not quickly reversible [34]. Also, it is reported that the toxic effect of organophosphates is caused by phosphorylation of AChE in erythrocytes, nerve tissue, serum, and liver [13]. Use of 1 mL of corn oil containing 100 mg malathion/kg BW of rat per day had lower AChE activity than the control group [79]. Rezg et al. [79] found a marked increase in Hb concentration which can be considered as an adaptive method in order to supply more oxygen in response to pulmonary hurt induced by subchronic exposure to malathion that is inconsistent with our reports. The RBC, Hb, MCV, and MCHC were similar in rats fed with 60 mg sumithion/kg BW/day compared to the control group without pesticide. Anemia was observed about 1 month after administration of terbufos in the cattle [14]. It is reported that malathion caused severe hepatic and renal damages which increased levels of liver enzymes [SGOT, SGPT, alkaline phosphatase (ALP) and acid phosphatase (ACP)] and made many changes in kidney output including significantly increased levels of creatinine, urea and uric acid, and reduced total protein and albumin in the plasma of rats exposed to malathion at a dose level of 20 mg/kg BW [4]. Increasing of SGPT and SGOT [4] is consistent with our experiment after phosalone administration. In this study, although the level of AChE inhibitory activity (17.99%, mean of period 1 and 2) was markedly increased some blood indices such as Hb (9.66%, mean of period 1 and 2), RBC (12.45%), and MCHC (10.2%) were significantly decreased in ewes exposed to phosalone compared to the control group. These results are in line with different previous studies which indicated that the exposure to malathion and other pesticides cause to impel acute physiological and biochemical disorders in experimental animals, buffalo calves [86], goats [50], mice [63], cockerels [87], poultry [43], rabbits [101], and rats [2, 42]. Some of the harmful impacts of oxidative stress leading to the production of free radicals are due to the activity of a number of pesticides known as OP having a tendency to produce of oxidative stress [4]. Nevertheless, several studies showed that malathion impelled lipid peroxidation and oxidative stress in experimental animals [22, 32]. Dimethoate pesticide at a dose of 75 mg/kg BW inhibited AChE activity in Albino rat males [8]. A dose of 0.1 mg malathion/mouse caused a significant decline in RBC, leukocyte, Hb, neutrophil, eosinophil and monocytes contents, and an increase in reticulocytes [12]. Similar changes in RBC and Hb [92] have been found using different pesticides and various experimental methods. In the present study among measured plasma metabolites, AChE inhibitory activity, SGPT, and SGOT is affected by the phosalone feeding. The Hb, RBC, and MCHC of hematology parameters were also decreased with the addition of phosalone to the diet. AChE activity in other studies of human [78], carp [90], mice [59] exposed to organophosphate pesticides was significantly lower than the control group. Levels of blood AChE were markedly depressed only in the animals fed on silage from corn treated at a level of 907 g/0.4 ha, and the common health of the animals and their milk production did not seem to be affected by pesticide [15]. In comparison with other adsorbents to binding toxins [94], it is concluded that bentonites have unique characteristics followed by kaolin-pectin in affinity of toxins (Kaopectate). There was no significant effect on glucose, beta globulin, cholesterol, and total protein, but alpha globulin, gamma globulin, albumin and urea were affected by the treatment of 2% bentonite rather than control group [54]. Despite an increase in albumin (present study), an increase in total protein and a decrease in BUN of plasma can be related to the fact that bentonite application can form a steady state in the rumen following the absorption of ammonia, which alternatively ammonia will be released after the decrease in the pool of the rumen, hence the animals will have a more chance in microbial protein synthesis in the rumen [54]. In the present work, all of the hematology parameters and plasma metabolites not affected by SB supplementation was compared to the control group (P < 0.05).

Phosalone residues

Phosalone residue (mg/kg/day) in the milk of P2 treatment was approximately 11.45 and 11.26 (period 1 and 2, respectively) times more than P3 group. During period 1 and 2, excretion of phosalone via urine and faecal (mg/kg DM) for P3 group was higher than P2. The phosalone excreted via the urine and faecal (mg/kg DM) in the P3 group was approximately 1.65 and 1.06 (period 1 and 2) times more than P2 group, respectively. Fenthion at low levels (0.006-0.014 m/kg) was found only in the milk of cows fed on contaminated corn silage. The residues were also found in the urine (0.004-0.160 ppm) and feces (0.003-0.156 mg/kg, wet basis) of all cows following consuming of fenthion in the diets [15]. In this study, only 0.34 (period 1) and 0.36% (period 2) of phosalone residue is found in the feces of the P2 group following daily consumption of it. The concentration of phosalone in the milk of ewes was (0.630 and 0.608 vs. 0.055 and 0.054 mg/kg for P2 vs. P3 in period 1 and 2, respectively) above the maximum residue limit (MRL) of 0.01 mg/kg suggested by the European Union [27]. Therefore, use of ewe’s milk by humans can have negative effects on their health in the future. The JMPR [44] established an ADI of 0-0.02 mg/kg BW for phosalone. So by considering ADI at 0.02 mg/kg BW, health condition and immune response in persons with 60 kg BW who received 1.2 mg of phosalone/day may not be concerned during a short term. Also the concentration of phosalone in the body of a 60 kg person following consumption 1 kg milk from P2 or P3 treatment will be 0.619 or 0.054 (mean period 1 and 2) mg/day, respectively that both values are less than 1.2 mg, so milk consumption of this study will not be worrying for a 60 kg person in the short term. In Brazil, 30 milk samples and all components of the animals’ diet were tested for pesticide residues. From 30 milk samples, six (20%) were polluted with organophosphates, five (16.7%) with carbamate, and one illustration with both organophosphate and carbamate. From 48 tested feed cases, 15 (31.25%) were polluted to organophosphates, six (12.50%) to carbamate, and one case was contaminated with both pesticides [28]. Phosalone residues and their oxygen analogue were measured in the milk and tissues of dairy cows after administration at doses of 100, 200 and 500 mg/kg diet for 28 days [21]. The maximum phosalone residues with their metabolites at the 100 mg/kg of diet were about 0.03 mg/kg in the fat, 0.3 mg/kg in the liver, 0.05 mg/kg in the kidneys, 0.007 mg/kg in the milk and <0.05 mg/kg in other tissues. Feeding a goat with 957 mg phosalone resulted in excretion of 0.86 mg/kg in the milk, 0.27 mg/kg in the muscle, 1.02 mg/kg in the fat, 12.6 mg/kg in the liver and 13.1 mg/kg in the liver [100]. Chlorpyrifos as an OP fed in dairy cattle ration for 2 consecutive weeks by McKellar et al. [61]. The chlorpyrifos residues and their (mainly oxidized and hydroxylated) metabolites were observed at low doses in milk, and these residues decreased quickly after cessation of application. A similar finding by Johnson et al. [45] was reported. There were no diazinon residues in the milk of dairy cows following its feeding with a protein supplement [58]. Low levels of diazinon (<0.025–1 mg/kg) were found in the milk of cattle and sheep after consumption [71, 91]. Studies about application of SB for adsorption of OPs after administration is scarce, but the adsorption efficiency of SB for other toxins material in different animal discussed extensively {study of improvement of some serum biochemical changes in the broiler chickens administered 2.5 mg aflatoxin/kg diet by Kececi et al. [53]; the use of bentonite in the diet contaminated with aflatoxin, mainly decreased the unfavorable impacts of the aflatoxin in the rat [1]; using the specific clays (such as bentonite) can seriously diminished some of the negative effects associated with administration of aflatoxin to weanling pigs [83]; application of SB at 1.2% of ration showed good potential as aflatoxin binder and contamination of milk decreased up to 61% in dairy cows [23]}. In the present study, the concentration of phosalone in the milk samples of animals decreased during feeding SB in the P3 group, which indicated that it could not pass readily from the feed or digestive tract to the milk. The calcium bentonites considered as adsorbent clay (due to their surface charge and surface area) with high quality made of silicates or aluminosilicates. Results indicated that most of the calcium bentonites will often adsorb up to 100% of their dry weight of water and 80% oil [46, 66]. Toxins can be conducted into the porous structure of clay via electric elementary charges. The rate of adsorption can be affected by variable parameters such as size and the electric charge of the toxin or structure of clay [46]. Furthermore, micro-elements in the diet can be adsorbed by aluminosilicates and have deleterious impacts on the bioavailability of them in the body of animals [64, 99].


Using phosalone residues in milk may have unfavorable effects on the health of humans. When ewes fed on 280 mg phosalone per day for 63 consecutive days, pesticide residue found in milk, feces, and urine during both sampling periods. Supplementation of SB at 32 g/sheep/day resulted in significant reduction in phosalone content of milk, and increased milk yield and milk fat content. Also excretion of phosalone via urine and faecal significantly increased by application of SB in the ration. The present study indicates that feeding of phosalone to ewes caused a significant increase in AChE inhibitory activity and a significant reduction in RBC, Hb as well as MCHC. Although there was no significant change in DMI and BW gain following the feeding of both phosalone and SB together or alone in the animals, further studies are needed on a large number of animals.





Acid phosphatase


Acid detergent fiber


Acceptable daily intake


Acid insoluble ash


Alkaline phosphatase


Alanine transaminase


Aspartate Aminotransferase


Blood urea nitrogen


Body Weight




Crude protein


Film Thickness (μm)


Dry matter


Dry matter intake


For example


Ether extract




Gas chromatography–mass spectrometry






Mean corpuscular hemoglobin


Mean corpuscular hemoglobin concentration


Mean corpuscular volume










Maximum residue limit


Metabolic weight


Neutral detergent fiber




Organic matter


Organophosphate pesticide


Organophosphate pesticides




Packed cell volume


Part per milion


Red blood cell


Sodium bentonite


Serum glutamic oxaloacetic transaminase


Serum glutamic pyruvic transaminase


Selected ion monitoring


Undegradable intake protein


Volatile fatty acids




White blood cell


White blood cells




  1. Abdel-Wahhab MA, Nada SA, Farag IM, Abbas NF, Amra HA. Potential protective effect of HSCAS and bentonite against dietary aflatoxicosis in rat: with special reference to chromosomal aberrations. Nat Toxins. 1998;6:211–8.

    Article  CAS  PubMed  Google Scholar 

  2. Adeniran OY, Fafunso MA, Adeyemi O, Lawal AO, Ologundudu A, Omonkhua AA. Biochemical effects of pesticides on serum and urinological system of rats. J Appl Sci. 2006;6:668–72.

    Article  CAS  Google Scholar 

  3. Aguilera-Sot JI, Ramirez RG, Arechiga CF, Mendez-Llorente F, Lopez-Carlos MA, Silva-Ramos JM, Rincon-Delgado RM, Duran-Roldan FM. Effect of feed additives in growing lambs fed diets containing wet brewers grains. Asian-Aust J Anim Sci. 2008;21:1425–34.

    Article  Google Scholar 

  4. Al-Attar AM. Physiological and histopathological investigations on the effects of α-lipoic acid in rats exposed to malathion. J Biomed Biotechnol. 2010;2010:1–8.

    Article  Google Scholar 

  5. Al-Qarawi AA, Mahmoud OM, Haroun EM, Sobaih MA, Adam SE. Comparative effects of diazinon and melathion in Najdi sheep. Vet Hum Toxicol. 1999;41:287–9.

    CAS  PubMed  Google Scholar 

  6. AOAC. Official Methods of Analysis. 17th ed. Association of Official Analytical Chemist: Washington; 2000.

    Google Scholar 

  7. AOAC. Official Methods of 2007.01. 18th ed. Pesticide residues in foods by acetonitrile extraction and partitioning with magnesium sulfate gas chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. AOAC International: Gaithersburg, MD. 2007.

  8. Attia AA, Nasr HM. Dimethoate-induced changes in biochemical parameters of experimental rat serum and its neutralization by black seed (Nigella sativa L.) oil. Slovak J Anim Sci. 2009;42:87–94.

    Google Scholar 

  9. Barker MH, Sortwell RJ. Phosalone (11974 RP), Potential tumorigenic and toxic effects in prolonged dietary administration to rat. 1993. Accessed 24 Sep 2016.

    Google Scholar 

  10. Bates RL, Jackson JA. Glossary of geology. 2rd ed. Virginia: American Geological Institute; 1980.

    Google Scholar 

  11. Berthiaume R, Ivan M, Lafreniere C. Effects of sodium bentonite supplements on growth performance of feedlot steers fed direct-cut or wilted grass silage based diets. Can J Anim Sci. 2007;87:631–8.

    Article  CAS  Google Scholar 

  12. Bhatia A, Makkar M, Sohal N. Effect of subchronic and sublethal doses of malathion on some haematological and immunological parameters. Int J Environ Stud. 1996;51:59–66.

    Article  Google Scholar 

  13. Blood DC, Radostits OM, Gay CC. Veterinary Medicine. 8rd ed. London: Bailliere Tindall; 1994.

    Google Scholar 

  14. Boermans HJ, Black WD, Chesney J, Robb R, Shewfelt W. Effect of terbufos poisoning on the cholinesterase and hematological values in a dairy herd. Can Vet J. 1985;26:350–3.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Bowman MC, Leuck DB, Johnson JC, Knox FE. Residues of Fenthion in corn silage and effects of feeding dairy cows the treated silage. J Econom Entomol. 1970;63:1523–8.

    Article  CAS  Google Scholar 

  16. Britton RA, Colling DP, Klopfenstein TJ. Effect of complexing sodium bentonite with soybean meal or urea in vitro ruminal ammonia release and nitrogen utilization in ruminants. J Anim Sci. 1978;46:1738–47.

    Article  CAS  Google Scholar 

  17. Cobon DH, Stephenson RGA, Hopkins PS. The effect of oral administration of methionine, bentonite, methionine/bentonite and methionine/oil homogenates on wool production of grazing and penned sheep in a semi-arid tropical environment. Aust J Exp Agric. 1992;32:435–41.

    Article  CAS  Google Scholar 

  18. Colling DP, Britton RA, Farlin SD, Nielsen MK. Effects of adding sodium bentonite to high grain diets for ruminants. J Anim Sci. 1979;48:641–8.

    Article  CAS  Google Scholar 

  19. Colovic MB, Krstic DZ, Lazarevic-Pasti TD, Bondzic AM, Vasic VM. Acetylcholinesterase Inhibitors: Pharmacology and Toxicology. Curr Neuropharmacol. 2013;11:315–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Cook RM, Wilson KA. Removal of Pesticide Residues from Dairy Cattle. J Dairy Sci. 1971;54:712–8.

    Article  CAS  PubMed  Google Scholar 

  21. Craig L, Landknecht J, Adams L, Busemeyer F, Bache B. Residue determination of phosalone and its oxygen analog in the milk and tissues of dairy cattle by electron capture gas chromatography. 1980. Accessed 10 Sep 2016.

    Google Scholar 

  22. Da Silva AP, Farina M, Franco JL, Dafre AL, Kassa J, Kuca K. Temporal effects of newly developed oximes (K027, K048) on malathion-induced acetylcholinesterase inhibition and lipid peroxidation in mouse prefrontal cortex. Neurotoxicology. 2008;29:184–9.

    Article  CAS  PubMed  Google Scholar 

  23. Diaz DE, Hagler WM, Blackwelder JT, Eve JA, Hopkins BA, Andersen KL, Jones FT, Whitlow LW. Aflatoxin binders II: reduction of aflatoxin M1 in milk by sequestering agents of cows consuming aflatoxin in feed. Mycopathologia. 2004;157:233–41.

    Article  CAS  PubMed  Google Scholar 

  24. Dunn BH, Emerick RJ, Embry LB. Sodium bentonite and sodium bicarbonate in high-concentrate diets for lambs and steers. J Anim Sci. 1979;48:764–9.

    Article  CAS  Google Scholar 

  25. Ehrlich WK, Davison TM. Adding bentonite to sorghum grain-based supplements has no effect on cow milk production. Aust J Exp Agric. 1997;37:505–8.

    Article  Google Scholar 

  26. Elman GL, Courney KD, Andres V, Featherstone RMA. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88–95.

    Article  Google Scholar 

  27. EU Pesticides database. Maximum Residue Levels, Reg. (EU) No 899/2012. 2013. Accessed 13 Sep 2016.

  28. Fagnani R, Beloti V, Battaglini APP, Dunga KS, Tamanini R. Organophosphorus and carbamates residues in milk and feedstuff supplied to dairy cattle. Pesq Vet Bras. 2011;31:598–602.

    Article  Google Scholar 

  29. FAOSTAT. Food and Agriculture Organization of the United Nations Statistics Division. 2013. Accessed 16 Sep 2016.

    Google Scholar 

  30. Fenn PD, Leng RA. Wool growth and sulfur amino acid entry rate in sheep fed roughage based diets supplemented with bentonite and sulfur amino acids. Aust J Agric Res. 1989;40:889–96.

    Article  CAS  Google Scholar 

  31. Fenn PD, Leng RA. The effect of bentonite supplementation on ruminal protozoa density and wool growth in sheep either fed roughage based diets or grazing. Aust J Agric Res. 1990;41:167–74.

    Article  Google Scholar 

  32. Fortunato JJ, Agostinho FR, Reus GZ, Petronilho FC, Dal-Pizzol F, Quevedo J. Lipid peroxidative damage on malathion exposure in rats. Neurotox Res. 2006;9:23–8.

    Article  CAS  PubMed  Google Scholar 

  33. Gilden RC, Huffling K, Sattler B. Pesticides and health risks. J Obstet Gynecol Neonatal Nurs. 2010;39:103–10.

    Article  PubMed  Google Scholar 

  34. Goel A, Aggarwal P. Pesticide poisoning. Natl Med J India. 2007;20:182–91.

    PubMed  Google Scholar 

  35. Greenman SB, Rutten MJ, Fowler WM, Scheffer L, Shortridge LA, Brown B, Sheppard BC, Devoney KE, Devoney CW, Trunkey DD. Herbicide/pesticide effects on intestinal epithelial growth. Environ Res. 1997;75:85–93.

    Article  CAS  PubMed  Google Scholar 

  36. Harlin KS, Dellinger JA. Retina, brain and blood cholinesterase levels in cats treated with oral dichlorvos. Vet Hum Toxicol. 1993;35:201–3.

    CAS  PubMed  Google Scholar 

  37. Hatjian BA, Mutch E, Williams FM, Blain PG, Edwards JW. Cytogenetic response without changes in peripheral cholinesterase enzymes following exposure to a sheep dip containing diazinon in vivo and in vitro. Mutat Res. 2000;472:85–92.

    Article  CAS  PubMed  Google Scholar 

  38. Hirosawa N, Ueyama J, Kondo T, Kamijima M, Takagi K, Fujinaka S, Hirate A, Hasegawa T, Wakusawa S. Effect of DDVP on urinary excretion levels of pyrethroid metabolite 3-phenoxybenzoic acid in rats. Toxicol Lett. 2011;203:28–32.

    Article  CAS  PubMed  Google Scholar 

  39. Hristova AN, Ivan M, Neill L, McAllister TA. Evaluation of several potential bioactive agents for reducing protozoal activity in vitro. Anim Feed Sci Technol. 2003;105:163–84.

    Article  Google Scholar 

  40. Ivan M, Dayrell MS, Hidiroglou M. Effects of bentonite and monensin on selected elements in stomach and liver of fauna-free and faunated sheep. J Dairy Sci. 1992;75:201–8.

    Article  CAS  PubMed  Google Scholar 

  41. Ivan M, Dayrell MS, Mahadevan S, Hidiroglou M. Effects of bentonite on wool growth and nitrogen metabolism in fauna-free and faunated sheep. J Anim Sci. 1992;70:3194–202.

    Article  CAS  PubMed  Google Scholar 

  42. Jabbar A, Khawaja SA, Iqbal A, Malik SA. Effect of malathion and methyl-parathion on rat liver enzymes. J Pak Med Assoc. 1990;40:266–70.

    CAS  PubMed  Google Scholar 

  43. Jayasree U, Gopala Reddy A, Reddy KS, Kalakumar B. Study on the mechanism of toxicity of deltamethrin in poultry. Ind J Toxicol. 2003;10:111–4.

    CAS  Google Scholar 

  44. JMPR. Pesticide residues in food. Report of the JMPR. FAO Plant Production and Protection. 2001. Accessed 28 Sep 2016.

    Google Scholar 

  45. Johnson JC, Jones RL, Leuck DB, Bowman MC, Knox FE. Persistence of chlorpyrifos-methyl in corn silage and effects of feeding dairy cows the treated silage. J Dairy Sci. 1974;57:1467–73.

    Article  CAS  PubMed  Google Scholar 

  46. Jouany JP. Methods for preventing, decontaminating and minimizing the toxicity of mycotoxins in feeds. Anim Feed Sci Technol. 2007;137:342–62.

    Article  CAS  Google Scholar 

  47. Kabak B, Dobson AD, Var I. Strategies to prevent mycotoxin contamination of food and animal feed: a review. Crit Rev Food Sci Nutr. 2006;46:593–619.

    Article  CAS  PubMed  Google Scholar 

  48. Kashani Motlagh MM, Amiri Rigi Z, Yuzbashi AA. To evaluate an acid activated bentonite from Khorasan (Iran) for use as bleaching clay. Int J Eng Sci. 2008;19:83–7.

    Google Scholar 

  49. Katsiotia M, Katsiotisb N, Rounia G, Bakirtzisc D, Loizidoua M. The effect of bentonite/cement mortar for the stabilization/solidification of sewage sludge containing heavy metals. Cem Concr Compos. 2008;30(10):1013–9.

    Article  Google Scholar 

  50. Kaur H, Srivastava AK, Garg SK, Prakash D. Subacute oral toxicity of chlorpyriphos in goats with particular reference to blood biochemical and pathomorpholigical alteration. Ind J Toxicol. 2000;7:83–90.

    Google Scholar 

  51. Kaya H, Celik ES, Yilmaz S, Tulgar A, Akbulut M, Demir N. Hematological, serum biochemical, and immunological responses in common carp (Cyprinus carpio) exposed to phosalone. Comp Clin Pathol. 2015;24:497–507.

    Article  CAS  Google Scholar 

  52. Kazemi M, Tahmasbi AM, Valizadeh R, Naserian AA, Soni A. Organophosphate pesticides: A general review. Agric Sci Res J. 2012;2:512–22.

    Google Scholar 

  53. Kececi T, Oguz H, Kurtoglu V, Demet O. Effects of polyvinylpolypyrrolidone, synthetic zeolite and bentonite on serum biochemical and haematological characters of broiler chickens during aflatoxicosis. Br Poult Sci. 1998;39:452–8.

    Article  CAS  PubMed  Google Scholar 

  54. Khadem AA, Soofizadeh M, Afzalzadeh A. Productivity, blood metabolites and carcass characteristics of fattening Zandi lambs fed sodium bentonite supplemented total mixed rations. Pak J Biol Sci. 2007;10:3613–9.

    Article  CAS  PubMed  Google Scholar 

  55. Khanedar F, Vakili R, Zakizadeh S. Effects of two kinds of bentonite on performance, blood biochemical parameters, carcass characteristics and tibia ash of broiler chickens. Iran J Appl Anim Sci. 2013;3:577–81.

    CAS  Google Scholar 

  56. Kolosova A, Stroka J. Substances for reduction of the contamination of feed by mycotoxins: A review. World Mycotoxin J. 2011;4:225–56.

    Article  CAS  Google Scholar 

  57. Kumar V, Puniya M, Roy D. Effect of lignosulfonate supplementation on carryover of monocrotophos to milk in lactating crossbred goats. Ind J Dairy Sci. 2015;68:239–46.

    Google Scholar 

  58. Lloyd JE, Matthysse JG. Residues of dichlorvos, diazinon, and dimetilan in milk of cows fed PVC insecticide feed additives. J Econ Entomol. 1971;4:821–2.

    Article  Google Scholar 

  59. Long SM, Dawson A, Shore RF. A comparison of the effects of single and repeated exposure to an organophosphate insecticide on acetylcholinesterase activity in mammals. Environ Toxicol Chem. 2006;25:1857–63.

    Article  CAS  PubMed  Google Scholar 

  60. Mavrogenis AP, Papachristoforou C. Estimation of the energy value of milk and prediction of fat-corrected milk yield in sheep and goats. Small Rumin Res. 1988;1:229–36.

    Article  Google Scholar 

  61. McKellar RL, Dishburger HJ, Rice JR, Craig LF, Pennington J. Residues of chlorpyrifos, its oxygen analogue, and 3,5,6-trichloro-2-pyridinol in milk and cream from cows fed chlorpyrifos. J Agric Food Chem. 1976;24:283–6.

    Article  CAS  PubMed  Google Scholar 

  62. Merhi M, Raynal H, Cahuzac E, Vinson F, Cravedi JP, Gamet-Payrastre L. Occupational exposure to pesticides and risk of hematopoietic cancers: meta-analysis of case–control studies. Cancer Causes Control. 2007;18:1209–26.

    Article  CAS  PubMed  Google Scholar 

  63. Mohssen M. Biochemical and histopathological changes in serum creatinine and kidney induced by inhalation of Thimet (Phorate) in male Swiss albino mouse, Mus musculus. Environ Res. 2001;87:31–6.

    Article  CAS  PubMed  Google Scholar 

  64. Moshtaghian J, Parsons CM, Leeper RW, Harrison PC, Koelkeberck KW. Effect of sodium aluminosilicate on phosphorus utilization by chicks and laying hens. Poult Sci. 1991;70:955–62.

    Article  CAS  PubMed  Google Scholar 

  65. Mukheijee PK, Kumar V, Houghton PJ. Screening of Indian Medicinal Plants for Acetylcholinesterase Inhibitory Activity. Phytother Res. 2007;21:1142–5.

    Article  Google Scholar 

  66. Murray HH. Applied clay mineralogy, occurrences, processing and application of kaolins, bentonites, palygorskite-sepiolite, and common clays, Bentonite applications. 1st ed. Netherland: Elsevier; 2006. p. 111–30.

    Book  Google Scholar 

  67. Murray PJ, Rowe JB, Aitchison EM. The effect of bentonite on wool growth, live weight change and rumen fermentation in sheep. Aust J Exp Agric. 1990;30:39–42.

    Article  CAS  Google Scholar 

  68. Nag SK, Mahanta SK, Raikwar MK, Bhadoria BK. Residues in milk and production performance of goats following the intake of a pesticide (endosulfan). Small Rumin Res. 2007;67:235–42.

    Article  Google Scholar 

  69. Nag SK, Raikwar MK. Persistent organochlorine pesticide residues in animal feed. Environ Monit Assess. 2011;174:327–35.

    Article  CAS  PubMed  Google Scholar 

  70. Nageswara Rao SB, Chopra RC. Influence of sodium bentonite and activated charcoal on aflatoxin M1 excretion in milk of goats. Small Rumin Res. 2001;41:203–13.

    Article  Google Scholar 

  71. Naidenov NK, Savov AA, Petkova MG. Retention and excretion of the organophosphoric preparation of diazinon by sheep. Biologia et Immunologia Reproductionis. 1984;9:81–6.

    CAS  Google Scholar 

  72. NRC. Nutrient Requirements of Small Ruminants: Sheep, Goats, Cervids, and New World Camelids. 6rd ed. Washington: National Academy Press; 2007.

    Google Scholar 

  73. Peter JV, Sudarsan TI, Moran JL. Clinical features of organophosphate poisoning: A review of different classification systems and approaches. Ind J Crit Care Med. 2014;18:735–45.

    Google Scholar 

  74. Prasai TP, Walsh KB, Bhattarai SP, Midmore DJ, Van TTH, Moore RJ, Stanley D. Biochar, bentonite and zeolite supplemented feeding of layer chickens alters intestinal microbiota and reduces campylobacter load. PLoS One. 2016;11:e0154061.

    Article  PubMed  PubMed Central  Google Scholar 

  75. Proskocil BJ, Bruun DA, Thompson CM, Fryer A, Lein P. Organophosphorus pesticides decrease M2 muscarinic receptor function in guinea pig airway nerves via indirect mechanisms. PLoS One. 2010;5:e10562.

    Article  PubMed  PubMed Central  Google Scholar 

  76. Rajwa U, Sandhu KS. Effect of handling and processing on pesticide residues in food- a review. J Food Sci Technol. 2014;51:201–20.

    Article  Google Scholar 

  77. Reddy SJ, Reddy BV, Ramamurthi R. Effect of chronic insecticide, phosalone, toxicity on haem synthesis and blood gas composition in the rat. Biochem Int. 1992;26:551–8.

    CAS  PubMed  Google Scholar 

  78. Rendon von Osten J, Epomex C, Tinoco-Ojanguren R, Soares AM, Guilhermino L. Effect of pesticide exposure on acetylcholinesterase activity in subsistence farmers from Campeche, Mexico. Arch Environ Health. 2004;59:418–25.

    Article  PubMed  Google Scholar 

  79. Rezg R, Mornagui B, Kamoun A, El-Fazaa S, Gharbi N. Effect of subchronic exposure to malathion on metabolic parameters in the rat. C R Biol. 2007;330:143–7.

    Article  CAS  PubMed  Google Scholar 

  80. Rhodia, Inc. Results for phosalone residue in tissues of beef cattle and sheep concerning blood plasma. 1967. Accessed 10 Oct 2016.

    Google Scholar 

  81. Rhodia, Inc. Results for phosalone in milk of dairy cattle-Woodard Research Centre Project. 1967. Accessed 12 Oct 2016.

    Google Scholar 

  82. Sangha GK, Kaur K, Khera KS, Singh B. Toxicological effects of cypermethrin on female albino rats. Toxicol Int. 2011;18:5–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Schell TC, Lindemann MD, Kornegay ET, Blodgett DJ, Doem JA. Effectiveness of different types of clay for reducing the detrimental effects of aflatoxin-contaminated diets on performance and serum profiles of weanling pigs. J Anim Sci. 1993;71:1226–31.

    CAS  PubMed  Google Scholar 

  84. Senthil Kumar N, Chopra RC, Chhabra A, Wadhawa BK. Effect of activated charcoal against chlorpyrifhos on blood biochemical parameters in calves. Ind J Anim Nutr. 2007;24:190–2.

    Google Scholar 

  85. Sharma A, Gill JP, Bedi JS, Pooni PA. Monitoring of pesticide residues in human breast milk from Punjab, India and its correlation with health associated parameters. Bull Environ Contam Toxicol. 2014;93:465–71.

    Article  CAS  PubMed  Google Scholar 

  86. Singh G, Sharma LD, Ahmad AH, Singh SP. Fenvalerate induced dermal toxicity in buffalo calves. J Appl Anim Res. 1999;16:205–10.

    Article  CAS  Google Scholar 

  87. Singh G, Sharma LD, Singh SP, Ahmad AH. Haematobiochemical profiles in cockerels following prolong feeding of fenvalerate. Ind J Agron. 2003;8:141–5.

    Google Scholar 

  88. Stepan R, Ticha J, Hajslova J, Kovalczuk T, Kocourek V. Baby food production chain: pesticide residues in fresh apples and products. Food Addit Contam. 2005;22:1231–42.

    Article  CAS  PubMed  Google Scholar 

  89. Sugeng AJ, Beamer PI, Lutz EA, Rosales CB. Hazard-ranking of agricultural pesticides for chronic health effects in Yuma County, Arizona. Sci Total Environ. 2013;463–464:35–41.

    Article  PubMed  Google Scholar 

  90. Szabo A, Nemcsok J, Asztalos B, Rakonczay Z, Kasa P, Hieu LH. The effect of pesticides on carp (Cyprinus carpio L). Acetylcholinesterase and its biochemical characterization. Ecotoxicol Environ Saf. 1992;23:39–45.

    Article  CAS  PubMed  Google Scholar 

  91. Szerletics TM, Soos K, Vegh E. Determination of residues of pyrethroid and organophosphorus ectoparasiticides in foods of animal origin. Acta Vet Hung. 2000;48:139–49.

    Google Scholar 

  92. Szubartowska E, Gromgsz-Kalkowska K. Blood morphology in quails after poisoning with fenitrothion. Comp Biochem Physiol C Toxicol Pharmacol. 1992;101:263–7.

    Article  CAS  Google Scholar 

  93. Talebi K. Dissipation of phosalone and diazinon in fresh and dried alfalfa. J Environ Sci Health B. 2006;1:595–603.

    Article  Google Scholar 

  94. Trckova M, Matlova L, Dvorska L, Pavlik I. Kaolin, bentonite, and zeolites as feed supplements for animals: health advantages and risks. Vet Med. 2004;49:389–99.

    CAS  Google Scholar 

  95. Van Kuelen J, Young BA. Evaluation of acid-insoluble ash as a natural marker in ruminant digestibility studies. J Anim Sci. 1977;44:282–7.

    Article  Google Scholar 

  96. Van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber and non starch polysaccharides in ration to animal nutrition. J Dairy Sci. 1991;74:3583–97.

    Article  CAS  PubMed  Google Scholar 

  97. Wallace RJ, Newbold CJ. Effects of bentonite on fermentation in the rumen simulation technique (rusitec) and on rumen ciliate protozoa. J Agric Sci (Camb). 1991;116:163–8.

    Article  CAS  Google Scholar 

  98. Walz LS, White TW, Fernandez JM, Gentry LR, Blouin DC, Froetschel MA, Brown TF, Lupton CJ, Chapa AM. Effects of fish meal and sodium bentonite on daily gain, wool growth, carcass characteristics, and ruminal and blood characteristics of lambs fed concentrate diets. J Anim Sci. 1998;76:2025–31.

    Article  CAS  PubMed  Google Scholar 

  99. Ward TL, Watkins KL, Southern LL, Hoyt PG, French DD. Interactive effects of sodium zeolite-A and copper in growing swine: growth, and bone and tissue mineral concentrations. J Anim Sci. 1991;69:726–33.

    Article  CAS  PubMed  Google Scholar 

  100. Witkonton S, Wilkes L, Wargo J. Determination of phosalone and its oxygen analog in the milk and tissues of dairy cattle by electron capture gas chromatography. 1979. Accessed 11 Nov 2016.

    Google Scholar 

  101. Yousef MI, El-Demerdash FM, Kamel KI, Al-Salhen KS. Changes in some hematological and biochemical indices of rabbits induced by isoflavones and cypermethrin. Toxicology. 2003;189:223–34.

    Article  CAS  PubMed  Google Scholar 

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We thank to University of Torbat-e Jam, Ferdowsi University of Mashhad, Vivan Company and Mr. Morteza Kazemi.


Financial support of this project was conducted by University of Torbat-e Jam, Ferdowsi University of Mashhad, and Vivan Company.

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The authors were also involved in carrying out of this project. MK is the main designer of the project. Other authors were as consultant in this project. All authors read and approved the final manuscript.

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The authors declare that they have no competing interests.

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All experimental procedures in this project were approved by the Animal Use and Care Administrative Advisory Committee at Ferdowsi University of Mashhad. The protocol of this project was also reviewed and approved by Ferdowsi University of Mashhad. Animals were kept in Research Station of Faculty of Agriculture, Ferdowsi University of Mashhad.

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Kazemi, M., Torbaghan, A.E., Tahmasbi, A.M. et al. Effects of phosalone consumption via feeding with or without sodium bentonite on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep. J Anim Sci Technol 59, 10 (2017).

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