Parasitic infections caused by Eimeria species are responsible for most economic losses in poultry production. Prevalence studies can adequately assist the design of prophylaxis strategies for disease control. Therefore, stool samples from 251 flocks of broilers from 28 to 48 days old were collected in 21 municipalities in the state of Santa Catarina, Brazil, to detect and examine the prevalence of Eimeria acervulina, Eimeria maxima, Eimeria tenella, Eimeria mitis, Eimeria praecox, Eimeria necatrix, and Eimeria brunetti. The oocysts were recovered and quantified, and the species were identified by a multiplex PCR technique. Amplicons of seven Eimeria species originating from the PCR-positive samples were cloned. Microscopy studies demonstrated that 96% of the farms were positive for the Eimeria. Seven species were identified, as follows: E. maxima (63.7%) and E. acervulina (63.3%) were the most prevalent species, followed by E. tenella (54.6%), E. mitis (38.6%), E. praecox (25.1%), E. necatrix (24.3%), and E. brunetti (13.1%). The average number of species detected per farm was 2.96, and the most common were E. acervulina, E. maxima, and E. tenella (9.16%). The sequencing of the clones confirmed the specificity and effectiveness of multiplex PCR for the identification of seven species of Eimeria, so this tool can be useful in studying circulating species in poultry farms, thereby assisting prophylactic measures against coccidiosis.

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Phage Display is a powerful tool for identification of protein ligands against a myriad of targets. In our study, we selected a small protein member from the Kunitz family to serve as a scaffold in the MIII 13 phage coat protein. The protein, originally a scorpion toxin that inhibits a potassium‐dependent voltage channel, has 33 residues composed of two beta sheets and an alpha helix stabilized by disulfide bonds. After cloning the toxin gene into an M13 phagemid, we used Kunkel's mutagenesis, an additional uracil‐DNA glycosylase treatment, and rolling circle amplification (RCA) procedures to produce a synthetic Kunitz‐type phage display library. We drove the mutations in specific residues within two specific loops as well as within the alpha helix of the scaffold protein, keeping the cysteines intact. We were able to produce a library of over a million different proteins. To validate the ability of this library to bind specific targets, trypsin was used as target. Since other studies have proved Kunitz's natural proteins as protease inhibitors, we expect our displayed Phage protein library to generate new potent and specific protease inhibitors for medical as well as biotechnological purposes.Support or Funding InformationThis work was supported by “State University of Santa Catarina” (UDESC) and Fundação de Amparo a Pesquisa e Inovação do Estado de Santa Catarina (FAPESC).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.