Tissue distribution of artemisinin in broiler chickens following single or multiple oral administration

Background: artemisinin is commonly used for the treatment of malaria, but recently has been considered as a potential substance to control poultry coccidiosis

Objectives: the aim of the present study was to determine the tissue distribution of artemisinin following single or multiple oral administration of different doses in broiler chickens

Methods: a total number of 390 one day old Ross broiler chicks were divided randomly into two main groups, in the first group 0, 1, 5, 25, 125, 250, 500 or 1000 mg/kg artemisinin as a single oral dose was administered on day 44, but the second group were treated with 0, 17, 34, 68 or 136 ppm artemisinin from day 8 to day 44. The HPLC system was used to determine the level of artemisinin in different tissue samples. Data were assessed using one way analysis of variance [ANOVA] followed by the Tukey's test [p<0.05]

Results: maximum concentrations of artemisinin were found in the liver of chickens in both groups in a dose dependent manner. While, the minimum level was determined in the brain and the kidney of chickens received multiple artemisinin administration; in the spleen of those chickens a single oral dose was administered. The concentration of artemisinin in the brain reached a plateau at 68 ppm in multiple administrations and 125mg/kg at single dose, no shift was found with dose increment

Conclusions: it can be concluded that tissue accumulation of artemisinin is time and dose dependent. Moreover, redistribution, saturation effect and tissue selectivity were also observed

[ level of differentiation and function of the enteric nervous system in the chicken embryo]

Any developmental disorder in enteric nervous system [ENS] may lead to congenital motility diseases of the gastrointestinal tract. The present study aimed to examine the structural differentiation and functional activity of the ENS in the Chick embryo. Ten Chick embryos were sacrificed at embryonic day 19 and then, their jejunum and colon specimens were collected. The isolated rings of the intestine were prepared and their motor activity was tested in an organ bath system. The contraction of the tissues was recorded in basic condition and following the stimulation by cholinergic and adrenergic agonists as well as the stimulation of the non-adrenergic-non-cholinergic system by electrical field stimulation [EFS]. The structure of the intestinal specimen was assessed immuno- 1histochemically [IHC] using glial fibrillary acidic protein [GFAP] biomarker. Spontaneous rhythmic contractions were seen in both jejunum and colon specimens. Cholinergic stimulations significantly increased the amplitude of contractions in jejunum [p<0.01] and colon [p<0.001] tissues. However, adrenergic stimulation decreased significantly the amplitude of contractions in isolated tissues prepared from the jejunum [p<0.05] and colon [p<0.001]. The EFS-induced decreases significantly the tension of isolated tissues pre-contracted with potassium chloride in both jejunum [p<0.001] and colon [p<0.001]. The results of IHC were showed a positive immunoreactivity of enteric nervous ganglia with GFAPbiomarker. It seems that the ENS in chick intestine is fully differentiated before birth and it can control the intestinal motility patterns in birds

Blood and tissue levels of diazinon in rabbit following a subacute dermal exposure to incremental doses

Uncontrolled application of diazinon [DZN] can cause environmental contamination and adverse health effects on humans or animals. This study aimed to investigate the toxic effects and the level of DZN in serum and tissues of rabbits following a sub acute dermal exposure to toxicant. Different doses of DZN were applied daily to New Zealand rabbits through the ear skin in incremental doses for 4 weeks. Blood samples were collected at the beginning and the end of each dose-week period. Tissue samples were collected from brain, muscle, kidney and liver on day 28, after euthanizing the rabbits. DZN contents of the blood and tissue samples were measured using a reversed phase HPLC system. Clinical observations indicated signs of toxicity in the animals exposed to DZN as shown by diarrhea and body weight loss from day twenty. The level of DZN in the blood elevated with enhancing exposure time and reached the highest level at the end of the fourth week [0.620 +/- 0.26ppm]. The highest level of DZN was found in the brain tissue [0341 +/- 0.015 ppm]. The results of this study revealed the tissue accumulation and subsequent toxic effects of DZN following the subacute dermal exposure to the toxicant. It suggests that the determination of the toxicant level in the serum or tissue can be a monitoring method for the detection of the contamination rate

[Comparison between anticoccidial effect of granulated Artemisia siberi extract and pure Artemisinin in affected broilers]

Increasing resistance against conventional anticoccidial drugs and the consequence of their residues has paid the attention toward more effective and safe compounds. Artemisia plant is a potential candidate that its anticoccidial effect has been previously discussed. This study aimed to produce a granule from the extract of Artemisia siberi and evaluate its anticcocidial effects compared to pure Atremisinin. Artemisinin was extracted from Artemisia by petroleum ether and then formulated into a wet granule. Experimental coccidiosis was induced in chicks [n=75] by oral administration of 250000 oocysts/chick. Chicks were divided into 5 groups of three replicates each [n=15] and one uninfected group [n=15]. The infected chicks were treated by oral administration [lmg/kg] of pure artemisinin and granule formulation with three different doses [1, 2.5 and 5mg/kg artemisinin] as feed additive. The treatment was conducted for 5 successive days towever. The fifth infected group and uninfected group did not receive any medication. At the end of treatment, fecal samples of each group were collected for 5 days and the OPG [oocyst per gram] was determined as anticoccidial index. The granule formulation of Artemisia and pure artemisinin significantly [p<0.001] decreased the OPG values in treated groups [30% in treated groups Vs8% in control]. However, there wasn't significant difference between granule formulation and pure artemisinin on OPG reduction [30.39% and 30.35%, respectively]. This study showed that the Artemisia siberi granulated extract can be considered as a new effective and safe anticoccidial drug

[Bioequivalence study of two closantel formulations in sheep]

Bioequivalence study is a scientific and practical method used to compare the quality of a generic drug with a reference product. This study aimed to examine the bioequivalence of two closantel formulations produced with different sources of raw material by a domestic pharmaceutical company. Due to long half-life of closantel, the study carried out by a parallel method. Thirty sheep were divided into 2 groups of 15 each. In the first group [test group] each sheep received 500 mg bolus of closantel and in the second group [reference group], each sheep received a 500 mg bolus of closantel produced with a raw material from a Belgium Company. Blood samples were collected at 0, 4, 8, 12, 16, 20, 24, 32, 48 and 72 hours after drug administration. An HPLC system was used to determine the amount of closantel in plasma. Pharmacokinetic parameters including area under curve [AUC], C[max], T[max], Kel and t[1/2] of closantel were determined in each sheep. A t-student test was used to analyze and compare the results. The mean +/- SD of C[max] in reference and test groups were 56.38 +/- 14.28 micro g/ml, 51.44 +/- 10.55, respectively. T[max] in reference and test groups were 22.93 +/- 2.81, 23.72 +/- 1.83 h, respectively. AUC [0-72] in reference and test drugs were 2049.1 +/- 421.2 and, 1795.1 +/- 421.2, respectively. AUC [0- infinity] in reference and test were 2557.3 +/- 621.5 and 2171.1 +/- 80.K[el] in reference group was 0.0321 +/- 0.0144 while in test it was 0.0348 +/- 0.0128. t[1/2] in reference and test groups were 24.85 +/- 8.34 h and, 22.29 +/- 8.03 h, respectively. In conclusion, the results of this study showed that there was not significant difference between reference and test group, suggesting that two products were bioequivalent

[Polyclonal antibody production against bovine serum albumin conjugated artemisinin in rabbit]

The aim of the present study was to produce a polyclonal antibody against bovine serum albumin [BSA] conjugated with artemisinin. To gain an immunogenic character of artemisinin, a carboxyl group was added to it using mixed anhydride method. Then, the reactive compound of artemisinin was conjugated with BSA. The BSA+artemisinin were injected to white female New Zealand rabbits for two times. In the first injection, the Fraud's complete adjuvant was used, and two weeks later, a booster injection was carried out with a Fraud's incomplete adjuvant. Two weeks later, blood samples were collected; their serum were separated and frozen until assessment. The production of antibody against BSA+artemisinin was assayed by Immunoblot technique. Antibody was separated and concentrated by saturated ammonium sulfate. The assay confirmed the successful production of an antibody against BSA+artemisinin as a fundamental step in investigation of pharmacodynamics and pharmacokinetics of artemisinin

Biochemical, hematological and pathological alterations associated with ascites in broilers and the role of oxygen-derived free radicals

Three hundred and fifteen day -old chickens were used to test the relationship between oxygen- derived free radicals and the biochemical, hematological and pathological alterations[associtd with ascites]. They were randomly divided into three experimental groups and ascites were developed in two groups of animals by exposing them to low temperature or administration of triiodothyronine [T[3],], and the third group was used as control. Different hematological, biochemical and pathological tests were used to determine the incidence of ascites in birds. These include total red blood cell [RBC], hematochrit [PCV], activities of alanine transaminase [ALT] and aspartate transaminase [AST] and the ratio of right ventricular weight to total ventricular weight [RV/TV]. Two hydroxylated salicylic acid [SA] metabolites, 2, 3- and 2, 5-dihydroxy benzoic acids [2, 3- and 2, 5-DHBA], were measured. by HPLC system to detect the generation of hydroxyl [OH] radicals. An analysis of variance [ANOVA] was used to determine the differences between different experimental groups. Ascites syndrome was observed in T[3] and low temperature treated groups as shown by necropsy changes and significant increases [p < 0.05] in the amount of RBC, PCV, ALT, AST and the ratio of RV/TV. While the significant increase was shown in the amounts of 2,3- and 2,5-DHBAfrom day 11, the alteration in the values of enzymes and hematoloic parameters and ratio of RV/TV occurred from days 18, 25 and 32 respectively. It can be concluded that OH radicals may be involved in the initiation of ascites syndrome, but the biochemical, hematological and pathological changes induced by these agents, can cause ascites and other alterations

[Simultaneous determination of salicylic acid and its hydroxylated metabolites in poultry serum by HPLC]

To introduce an HPLC method for indirect measuring of hydroxyl radicals in poultry serum. Chromatographic study. Poultry serum. By acidification of serum with 1.0 M HCl, salicylic acid [SA] and its hydroxylated metabolites were extracted into diethylether, dried and redissolved in mobile phase buffer and HCl. It was filtered and 20 micro L of the filtrate injected into the HPLC system. The chromatographic condition was as follows: acetate-citrate buffer [94%]: methanol [6%] as mobile phase run at 1.2 ml/min through a C18 column using UV and EC detectors concurrently. Descriptive statistics good. Separations among these compounds obtained without any interference with poultry serum elements. Calibration curves for SA and Dihydroxy benzoic acids [DHBAs] were linear at 0.28-88 micro g/ml [R[2] =0.996] and 0.3 -12.3 micro g/ml [R[2] = 0.998], respectively. Detection limit for SA and DHBAs were 0.56 micro g/ml and 0.3 micro g/ml, respectively. Precision of the method [CV%] as intra- and inter-day variations were defined for SA [2.5 -6.5%, 4.4-6.8%], 2,3-DHBA [4.2-8.5%, 5.8-9.0%] and 2, 5-DHBA [4.1-6.8%, 5.9- 7.4%]. Recovery of the method for SA and DHBA were 87 +/- 5% and 92 +/- 4%, respectively. This method is useful for simultaneous determination of SA and its hydroxylated metabolites in poultry serum as an indirect measurement of hydroxyl radicals