Simultaneous Analysis of Nicarbazin, Diclazuril, Toltrazuril, and Its Two Metabolites in Chicken Muscle and Eggs by In-Syringe Dispersive Solid-Phase Filter Clean-Up Followed by Liquid Chromatography-Tandem Mass Spectrometry.

Nicarbazin (NICA) and triazine anticoccidial drugs (diclazuril (DIZ) and toltrazuril (TOZ)) are the primary strategy for preventing and treating coccidiosis. To prevent the development of drug resistance and mitigate the potential chronic toxicity to humans resulting from prolonged exposure, a liquid chromatography-tandem mass spectrometry method with high reliability and sensitivity was developed to determine NICA, DIZ, TOZ, and its two metabolites in chicken muscle and eggs. Upon establishing the extraction conditions involving 10 mL of acetonitrile and 10 min of sonication, in-syringe dispersive solid-phase extraction with silica was performed in combination with n-hexane clean-up. The selection of isotope peaks of precursor ions and low-mass range scanning allowed the two transitions for the quantification of all compounds. The limits of detection for DIZ and NICA were both 0.1 µg/kg, and for TOZ and metabolites, they were 0.3 µg/kg; the limits of quantitation were 0.3 and 1 µg/kg, respectively. The linear range was 0.25-50 ng/mL with a correlation coefficient r > 0.999. The average recoveries at three spiking levels in muscle and eggs were 90.1-105.2% and 94.0-103.7% with the relative standard deviations of 3.0-8.1% and 3.1-14.4%, respectively. The precision, accuracy, and stability were evaluated by three quality control samples.

Age-dependent development trends (models) of intestinal significant microbiota species and Eimeria oocysts in coccidia-challenged broiler chickens as affected by dietary encapsulated organic acids and anticoccidial drugs.

RESEARCH HIGHLIGHTS: Cubic regression is the best model for explaining intestinal microbiota development.Polynomial regression is the best model for intestinal Eimeria oocysts development.Age-development trends are affected by dietary encapsulated organic acids and anticoccidials.

Quality control procedure for Coccidial vaccines versus different routes of immunization.

Background and Aim: Coccidiosis is an enteric infection caused by a protozoon (Eimeria tenella). Coccidiosis is known to have a negative impact on the economy. Coccidiosis is controlled using anticoccidial drugs, antibiotics, and vaccines. Various coccidial vaccines differ in application technique, attenuation method, and the species used. Coccidial vaccines can be spray or gel-based (Form). This study aimed to compare the effect of application and approaches between spray and gel vaccines for coccidiosis. Materials and Methods: Specific pathogen-free chicks were vaccinated with different vaccines. Fecal samples were taken on 21 days post-vaccination for vaccine take, and then a challenge test was done on day 21. Results: Post-vaccination oocyst counts in gel vaccinated groups were more than the spray vaccinated ones as it recorded (1400 and 2200) oocyst/g, but the gel vaccines resulted in lower post vaccinal titer which was (10000 and 12500) oocyst/g. Results of quantitative real-time polymerase chain reaction test post-vaccination were (23.72, 20.29) cycle threshold (CT) for spray vaccines and (18.75, 17.62) CT for gel vaccinated group. By challenging all the experimental groups, the microscopic and macroscopic lesion of gel vaccines resulted in score 1, while spray vaccines groups recorded score 2 and the control non-vaccinated challenged chickens showed score 4. The non-vaccinated/non-challenged group recorded a score of zero. Conclusion: These results can help poultry producers to decide which delivery system will provide the best results for their production system. The gel vaccines showed a better protection rate and lower shedding, which means more protection of birds and public health.

Hepatoprotective and renoprotective effects of silymarin against salinomycin-induced toxicity in adult rabbits.

Background and Aim: Salinomycin sodium, a licensed coccidiostat in rabbits, is used for fattening at a dose of 20-25 mg/kg. Salinomycin toxicity may arise from many risk factors (e.g., overdosage or use in non-target animal species). Silymarin extracted from milk thistle has antioxidant, anti-inflammatory, and antiviral properties. This study aimed to investigate the adverse impacts of oral administration of salinomycin for 28 consecutive days and how to reduce its risks and side effects by administering silymarin. Materials and Methods: Eighty-four male New Zealand White bucks (1.750-2.000 kg) were randomly divided into seven groups (12 each). Group one was the control. Groups two and three were administered salinomycin orally (doses of 20 and 40 mg/kg ration). Group four was administered salinomycin (20 mg/kg ration) and silymarin (6.5 mg/kg body weight [BW]). Group five received salinomycin (40 mg/kg ration) and silymarin (13 mg/kg BW). Groups six and seven were administered silymarin at doses of 6.5 and 13 mg/kg BW. Rabbits were euthanized and slaughtered on day 29 using the Halal method. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, urea, total proteins, albumin, total cholesterol, and high- and low-density lipoprotein (HDL and LDL) were analyzed in serum. Glutathione (GSH), superoxide dismutase (SOD), catalase, and malondialdehyde (MDA) were estimated in the liver. A histopathological investigation was performed on the liver and kidney. Results: The MDA activity, AST, ALT, total protein, albumin, total cholesterol, triglyceride, LDL, urea, and creatinine values were significantly elevated in groups two and three. The GSH, catalase, SOD, and HDL were significantly lower in these groups than in the control group. There were moderate pathologic changes in the liver and kidney of the third group. However, the results of the fourth and fifth groups improved more than those of the second and third groups. The results of the sixth and seventh groups were nearly the same as those of the control group. Conclusion: Salinomycin toxicity was caused by oxidative damage because of reactive oxygen species formation. Silymarin (6.5 or 13 mg/kg BW) tends to prevent and treat accidental toxicity. However, the high dose of silymarin (13 mg/kg BW) had more renal and hepatoprotective capacities.

Evaluation of sulfadimidine, amprolium and triquen to treat coccidiosis in wild pigeons / Avaliação da sulfadimidina, amprólio e triquenho no tratamento da coccidiose em pombos selvagens de Lower Dir, Khyber Pakhtunkhwa, Paquistão

Coccidiosis remains one of the major problems in poultry all over the world. Very limited data on anticoccidial drugs in wild pigeons is available. The current study was aimed to understand the comparative efficacy of sulfadimidine, amprolium and triquen in wild pigeons of Dir district, Pakistan suffering from coccidiosis. The faecal matter of wild pigeons were purchased from the local market for coccidian infection. Results revealed that 88.8% (16/18) were found infected with Eimeria spp. Three positive groups were treated with sulfadimidine (0.2mg/L), amprolium (25mg/L) and triquen. Sulfadimidine was most effective (45%) followed by amprolium (44.6%) while triquen (24.0%) showed less effectiveness against coccidiosis in pigeons. Number of oocysts were 79, 81 and 80 before treatment and 60, 44 and 44 after treatment with sulfadimidine, amprolium and triquen respectively. This study showed that sulphadimidine, amprolium and triquen could not significantly reduce the coccidiosis in pigeons. Further studies are required to clear the mechanism of anti-coccidial drugs in wild pigeons.

A coccidiose continua sendo um dos maiores problemas em aves de todo o mundo. Dados muito limitados sobre drogas anticoccidiais em pombos selvagens estão disponíveis. O presente estudo teve como objetivo compreender a eficácia comparativa da sulfadimidina, amprólio e triquenho em pombos selvagens do distrito de Dir, no Paquistão, que sofrem de coccidiose. A matéria fecal dos pombos selvagens foi comprada no mercado local para infecção por coccídios. Os resultados revelaram que 88,8% (16/18) foram encontrados infectados com Eimeria spp. Três grupos positivos foram tratados com sulfadimidina (0,2mg / L), amprólio (25mg / L) e triquenho. A sulfadimidina foi mais eficaz (45%), seguida pelo amprólio (44,6%), enquanto o triquenho (24,0%) apresentou menor efetividade contra a coccidiose em pombos. O número de oocistos foram 79, 81 e 80 antes do tratamento e 60, 44 e 44 após o tratamento com sulfadimidina, amprólio e triquenho, respectivamente. Este estudo mostrou que sulphadimidine, amprolium e triquen não poderiam reduzir significativamente a coccidiose em pombos. Mais estudos são necessários para limpar o mecanismo de drogas anticoccidiais em pombos selvagens.

Evaluation of sulfadimidine, amprolium and triquen to treat coccidiosis in wild pigeons / Avaliação da sulfadimidina, amprólio e triquenho no tratamento da coccidiose em pombos selvagens de Lower Dir, Khyber Pakhtunkhwa, Paquistão

Coccidiosis remains one of the major problems in poultry all over the world. Very limited data on anticoccidial drugs in wild pigeons is available. The current study was aimed to understand the comparative efficacy of sulfadimidine, amprolium and triquen in wild pigeons of Dir district, Pakistan suffering from coccidiosis. The faecal matter of wild pigeons were purchased from the local market for coccidian infection. Results revealed that 88.8% (16/18) were found infected with Eimeria spp. Three positive groups were treated with sulfadimidine (0.2mg/L), amprolium (25mg/L) and triquen. Sulfadimidine was most effective (45%) followed by amprolium (44.6%) while triquen (24.0%) showed less effectiveness against coccidiosis in pigeons. Number of oocysts were 79, 81 and 80 before treatment and 60, 44 and 44 after treatment with sulfadimidine, amprolium and triquen respectively. This study showed that sulphadimidine, amprolium and triquen could not significantly reduce the coccidiosis in pigeons. Further studies are required to clear the mechanism of anti-coccidial drugs in wild pigeons.

A coccidiose continua sendo um dos maiores problemas em aves de todo o mundo. Dados muito limitados sobre drogas anticoccidiais em pombos selvagens estão disponíveis. O presente estudo teve como objetivo compreender a eficácia comparativa da sulfadimidina, amprólio e triquenho em pombos selvagens do distrito de Dir, no Paquistão, que sofrem de coccidiose. A matéria fecal dos pombos selvagens foi comprada no mercado local para infecção por coccídios. Os resultados revelaram que 88,8% (16/18) foram encontrados infectados com Eimeria spp. Três grupos positivos foram tratados com sulfadimidina (0,2mg / L), amprólio (25mg / L) e triquenho. A sulfadimidina foi mais eficaz (45%), seguida pelo amprólio (44,6%), enquanto o triquenho (24,0%) apresentou menor efetividade contra a coccidiose em pombos. O número de oocistos foram 79, 81 e 80 antes do tratamento e 60, 44 e 44 após o tratamento com sulfadimidina, amprólio e triquenho, respectivamente. Este estudo mostrou que sulphadimidine, amprolium e triquen não poderiam reduzir significativamente a coccidiose em pombos. Mais estudos são necessários para limpar o mecanismo de drogas anticoccidiais em pombos selvagens.

Evaluation of sulfadimidine, amprolium and triquen to treat coccidiosis in wild pigeons

Abstract Coccidiosis remains one of the major problems in poultry all over the world. Very limited data on anticoccidial drugs in wild pigeons is available. The current study was aimed to understand the comparative efficacy of sulfadimidine, amprolium and triquen in wild pigeons of Dir district, Pakistan suffering from coccidiosis. The faecal matter of wild pigeons were purchased from the local market for coccidian infection. Results revealed that 88.8% (16/18) were found infected with Eimeria spp. Three positive groups were treated with sulfadimidine (0.2mg/L), amprolium (25mg/L) and triquen. Sulfadimidine was most effective (45%) followed by amprolium (44.6%) while triquen (24.0%) showed less effectiveness against coccidiosis in pigeons. Number of oocysts were 79, 81 and 80 before treatment and 60, 44 and 44 after treatment with sulfadimidine, amprolium and triquen respectively. This study showed that sulphadimidine, amprolium and triquen could not significantly reduce the coccidiosis in pigeons. Further studies are required to clear the mechanism of anti-coccidial drugs in wild pigeons.

Resumo A coccidiose continua sendo um dos maiores problemas em aves de todo o mundo. Dados muito limitados sobre drogas anticoccidiais em pombos selvagens estão disponíveis. O presente estudo teve como objetivo compreender a eficácia comparativa da sulfadimidina, amprólio e triquenho em pombos selvagens do distrito de Dir, no Paquistão, que sofrem de coccidiose. A matéria fecal dos pombos selvagens foi comprada no mercado local para infecção por coccídios. Os resultados revelaram que 88,8% (16/18) foram encontrados infectados com Eimeria spp. Três grupos positivos foram tratados com sulfadimidina (0,2mg / L), amprólio (25mg / L) e triquenho. A sulfadimidina foi mais eficaz (45%), seguida pelo amprólio (44,6%), enquanto o triquenho (24,0%) apresentou menor efetividade contra a coccidiose em pombos. O número de oocistos foram 79, 81 e 80 antes do tratamento e 60, 44 e 44 após o tratamento com sulfadimidina, amprólio e triquenho, respectivamente. Este estudo mostrou que sulphadimidine, amprolium e triquen não poderiam reduzir significativamente a coccidiose em pombos. Mais estudos são necessários para limpar o mecanismo de drogas anticoccidiais em pombos selvagens.

Molecular characterization and functional analysis of Eimeria tenella citrate synthase.

Chicken coccidiosis, caused by an obligate intracellular protozoan parasite of the genus Eimeria, is a major parasitic disease in the intensively reared poultry industry. Due to the widespread use of anticoccidial drugs, resistance has become an inevitable problem. In our previous study, Eimeria tenella citrate synthase (EtCS) was found to be up-expressed in two drug-resistant strains (diclazuril-resistant and maduramycin-resistant strains) compared to drug-sensitive strain by RNA sequence. In this study, we cloned and expressed EtCS and obtain its polyclonal antibodies. Quantitative real-time polymerase chain (qPCR) reactions and Western blots were used to analyze the transcription and translation levels of EtCS in sensitive and three drug-resistant strains. Compared with the sensitive strain, the transcription of EtCS was both significantly upregulated in diclazuril-resistant and maduramycin-resistant strains, but was not significantly different in salinomycin-resistant strain. No significant difference was seen in translation level in the three drug-resistant strains. Indirect immunofluorescence indicated that EtCS was mainly located in the cytoplasm of sporozoites except for posterior refractile bodies and in the cytoplasm and surface of merozoites. Anti-rEtCS antibody has inhibitory effects on E. tenella sporozoite invasion of DF-1 cells and the inhibition rate is more than 83%. Binding of the protein to chicken macrophage (HD11) cells was confirmed by immunofluorescence assays. When macrophages were treated with rEtCS, secretion of nitric oxide and cell proliferation of the macrophages were substantially reduced. These results showed that EtCS may be related to host cell invasion of E. tenella and involve in the development of E.tenella resistance to some drugs.