Tibial dyschondroplasia (TD) is a common skeletal disease occurred in growth plate of fast-growing broilers. Thiram is a sort of chemical used for pesticide and fungicide. The excessive use of thiram increased the threat to animal and human health. In this study, we aimed to investigate the therapeutic mechanism of chlorogenic acid (CGA) on thiram-induced tibial dyschondroplasia. Broiler chickens were divided into three different groups, e.g., control, TD, and CGA. CGA was administrated after the induction of TD from 4ᵗʰ day to 7ᵗʰ day. Biochemical analysis was performed to detect the content of calcium (Ca) and phosphorus (P). Histological changes and degradation of extracellular matrix were observed through hematoxylin-eosin (H & E) and Masson staining. To further determine the mechanism, TUNEL staining and western blot were also performed to detect the apoptosis changes in growth plate of all groups. The results showed the disproportionation of Ca and P content and upregulation of apoptosis during the development of TD. But, after the administration of CGA, the ratio of Ca:P was upregulated, and the apoptosis was also downregulated. The current study shows the toxic effect of thiram on chickens and suggests that CGA is associated with a mechanism that plays a significant role in apoptosis induced by thiram in poultry industry.

Tibial dyschondroplasia (TD) is most the common tibiotarsal bone disease in rapidly growing birds throughout the world. There is accumulating evidence that COX-2 abnormal expression in tibia plays an important role in TD progression. So, the regulation of COX-2 is an ever more appealing target for therapeutic intervention in TD. Astragaloside IV has an indispensable role in maintaining COX-2 expression in many diseases. So, we designed this study to use Astragaloside IV (AST-IV) against TD-affected chickens. A total of 180 Arbor Acres chickens were randomly divided in the control group, TD group, and Astr (AST-IV-treated chickens) group. During the experiment, mortality, feed conversion ratio, physiological changes, biochemical criterion, liver antioxidant enzymes, and gene expression of COX-2 were examined in all the chicken groups at various days. The results showed that AST-IV administration restored the growth performance and tibia lesions and decreased the mortality as compared with TD chickens. The biochemical criterion (ALP, AST, and ALT) of serum and liver antioxidant enzymes (SOD, GSH-Px, MDA, and T-AOC) improved after the administration of AST-IV. The COX-2 gene was upregulated significantly (P < 0.05) in TD chickens. Whereas, AST-IV treatment downregulated both gene and protein expression of COX-2 significantly (P < 0.05) in TD-affected chickens. AST-IV recovered tibial dyschondroplasia chickens by increasing the growth performance, ameliorating tibial cartilage damage, and decreasing COX-2 expression. In conclusion, AST-IV can be used to prevent thiram-induced TD in chickens.

Tetramethyl thiuram disulfide (thiram) is one of the important pesticides, which is extensively used in agriculture, but if it is combined with the cell membrane, then it causes membrane damage, bone morphogenic inactivation, and inhibited angiogenesis. Thiram has been considered a common cause of tibial dyschondrolplasia (TD) in various avian species, because it becomes the part of feed due to environmental contamination and its overuse in agriculture as pesticides or fungicide. However, there is no systematic study on the changes of the correlation indexes with toxic effect of the thiram in chickens. Therefore, we evaluated the toxic effects of thiram on growth performance of chickens, viscera organ index, pathological changes in tissue, and gene expression associated with osteoblast differentiation, vascularization, and tibial bone development. For this study, 1-day chickens (n = 300) were randomly distributed into two equal groups, control group (normal basal diet) and thiram group (adding thiram 40 mg/kg in basal diet). The result presented that thiram group chickens were looking unhealthy, lazy, and showing clinical symptoms like lameness. Thiram treatment significantly reduced the performance of chickens, liver index, and tibial length compared with control group. The toxic effect of thiram increased the visceral organ index (spleen and cardiac), tibia index, and TD severity considerably. It also increased serum Ca²⁺ and P³⁺ concentration and decreased tibial density compared to control chickens but the difference was not significant. Histopathology of tibia and liver showed that there were severe lesions due to toxic effect of thiram. Furthermore, HIF-1α and VEGF antibody localizations were increased and WNT4 localization was reduced significantly in immunohistochemical analysis. This systemic study of toxic effects of thiram in chicken concluded that thiram reduced the growth performance of chickens through decreasing liver index, whereas increasing kidney, cardiac, and spleen index, and induced TD by changing the expressions of VEGF, HIF-1α, and WNT4.