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It is estimated that nearly 1.8 billion people worldwide use pesticides to minimize the crop damage in efforts to increase food production to feed the growing world population. Exposures to lower concentartion of various pesticides such as fipronil and phenylpyrazole can result in fatal poisonings as well as airway inflammation. Inhalation of endotoxins, which are frequently present in the agricultural buildings, also induces lung inflammation. Animals exposed to both fipronil and endotoxin may demonstrate enhanced lung inflammation. There, however, are limited data available on the molecular mechanisms underlying the toxicity induced by fipronil alone or in combination with endotoxin. Hence, we employed a comprehensive microarray approach in a mouse model to identify the molecular mechanism involved in the lung inflammation by exposing the mice to fipronil (1/20th of LD50/d orally for 90 d) followed by intranasal E. coli LPS (80μg/mouse) or normal saline (N=10 each). Fipronil with or without LPS significantly (p<0.05) increased the total histopathological score suggesting lung damage. Ingenuity Pathway Analysis (IPA) software analysis predicted eicosanoid signalling among the top 5 canonical pathways involved in airway inflammation. Eicosanoid signalling plays important role in pulmonary inflammation and immune response via cytokine production, cell proliferation, antibody formation and antigen presentation. Fipronil alone caused 3‐fold increase in Phospholipase A2G5 (PLA2G5), which hydrolyses the membrane phospholipids to generate various eicosanoids. PLA2G5 showed 1.63 and 4.34 fold increase following exposure to endotoxin alone or in combination with fipronil, respectively. RT‐qPCR and immunohistochemistry analysis validated the microarray data for the PLA2G5. The data taken together indicate up‐regulation of functionally relevant PLA2G5 during fipronil or/and endotoxin induced lung inflammation, which may also play a role in lung inflammation associated with fipronil exposure.Support or Funding InformationGADVASU, LudhianaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Introduction: Virulent footrot of sheep caused by Dichelobacter nodosus is associated with tremendous economic losses due to recurrent treatment costs and increased culling rates. This organism being a fastidious anaerobe is difficult to isolate on ordinary media that does not support its growth. The D. nodosus serogroup B isolate described in the present study has been used in the preparation of the whole-cell killed vaccine against footrot in India. D. nodosus serogroup B is the predominant serogroup involved in virulent footrot (lesion score 4) in India as well as in many sheep-rearing countries of the globe. Methods: Genomic DNA was extracted using wizard Genomic DNA purification kit. The whole genome of the D. nodosus strain B was sequenced using an Illumina HiSeq 2500 platform and annotated according to functional gene categories. Annotations were performed using in-house developed Perl scripts using Nr/Nt database, uniprot, Pfam, KEGG, Panther DB, and GO database. Result: The assembled genome size is 1.311,533 Mb and GC content is 44.38. A total of 1215 protein-coding genes, 44tRNA and 7 rRNA were identified. The genome shows 98.63% sequence homology with the reference genome. However, 21 new genes have been identified in this genome. The information will provide insights into the various genes and regulators necessary for D. nodosus growth and survival. Discussion: The genome information of this serogroup B of D. nodosus isolate involved in 85–90% cases of virulent footrot of sheep in India provides further insights for improvement of the killed vaccine (B serogroup) developed recently in India. For the development of an efficacious vaccine against virulent footrot, it is essential to know the serological diversity as well as the virulent status of the strains of the D. nodosus. This serogroup isolate is a potential vaccine candidate to mitigate ovine footrot in India as the majority of virulent footrot cases belong to serogroup B of D. nodosus.