The agricultural sector uses fertilizers such as urea to add more nutrients to the soil needed for plant growth. Although it is cost-effective in crop production, indiscriminate use of nitrate-based fertilizer may result in behavioural, morphological, and physiological alterations on non-target organisms. This study determined the angiogenesis activity in the chorioallantoic membrane of urea-exposed duck embryos. It also investigated the weight, morphometries, and liver histopathology to gather more information on urea fertilizer's toxicity. It was observed that urea promoted angiogenesis in the CAM of duck embryos, especially at higher concentrations (P<0.05). Embryos treated with urea resulted in an alteration of the head-beak length (P<0.05). However, weight, crown-rump length, forelimb length, and hind limb length were not affected. The developing liver of urea-treated embryos showed distortion of the central vein shape and had larger sinusoidal spaces. The presence of Kupffer cells and lipid droplets were observed in the treated section. Congestion of blood cells, haemorrhage, and necrosis of hepatocytes were also observed in the tissue suggesting the extent of damage caused by the fertilizer. The findings of this study showed multiple developmental effects of urea on duck embryos. Further investigations are needed to shed more light on the toxicity of urea fertilizer on vertebrates.

Toll-like receptors (TLRs), type I transmembrane pattern recognition receptors (PRRs), are composed of the extracellular domain that is implicated in the recognition of microbial products and initiates the innate and adaptive immune response. Previous reports on TLRs in birds showed significant levels of inter- and intraspecific genetic variation. Little is known about the structure and function of the avian immune system, especially waterfowl species. This work aimed to identify and clone Anas platyrhynchos (mallard duck) TLR-3 (dTLR-3) and its expression level following challenge with velogenic Newcastle disease virus (NDV) as a model for waterfowl species. The mallard duck TLR-3 full-length cDNA sequence had been cloned, which consisted of 2457 nucleotides. The translated amino acid sequence showed identity degree as 97% with Muscovy duck, 95% with geese, 89% with helmeted guineafowls, 88% with the chickens TLR-3 gene, 82% with turkey TLR-3, and 79% with zebra finch, while it showed 54% with human one; the analysis data suggested that the new sequence is probably homologous to vertebrates’ TLR-3. The predicted protein encoded by the duck dTLR-3 mRNA sequence is composed of 819 amino acids. Analysis of the deduced amino acid sequence indicated that dTLR-3 has typical structural features and contains the main components of proteins in the TLR family. The dTLR-3 expressed in almost all examined tissues of mallard duck following quantitative real-time polymerase chain reaction (qPCR) analysis and using B-actin as a housekeeping gene. To check the functionality of the receptor and its role in viral infection, we evaluate the expression level in different tissues and its changes following NDV infection. The results showed significant (P < 0.05) upregulated in the brain at 24 h (1.84-fold), reached a peak at 48 h (4.82-fold), and recovered to normal levels at 72 h post-infection. These results indicate a complete and functional dTLR-3 that is orthologous to other vertebrate receptors with its potential role in early response against viral infection in mallard duck species.

Protection and restoration of different endangered bird species from pesticide exposure is crucial from the point of safety assessment of ecosystem. Toxicity predictions or risk assessment of pesticides by chemometric tools is one of the challenging fields in recent era. In the present study, classification-based quantitative structure toxicity relationship (QSTR) models were developed for a large dataset (516) of diverse pesticides on multiple avian species mallard duck, bobwhite quail, and zebra finch according to the Organization for Economic Co-operation and Development guidelines. The QSTR models were developed by linear discriminant analysis method with genetic algorithm for feature selection from 2D descriptors using QSAR-Co software. Different statistical metrics assured the reliability and robustness of the developed models. External compound prediction highlighted predictive nature of the models. The mechanistic interpretation suggested that presence of phosphate, halogens (Cl, Br), ether linkage, and NCOO influence the avian toxicity. Furthermore, model reliability was checked by the application of the standardization approach of the applicability domain (AD). Finally, the developed models provided a priori toxic and non-toxic classification for unknown pesticides (inside AD), with particular emphasis on organophosphate pesticides. The interspecies toxicity correlation and predictions encouraged for their further applicability for the fulfilment of data gaps in vital missing species.