Brachyspira hyodysenteriae and B. pilosicoli Proteins Recognized by Sera of Challenged Pigs

Abstract

The spirochetes Brachyspira hyodysenteriae and B. pilosicoli are pig intestinal pathogens that are the causative agents of swine dysentery (SD) and porcine intestinal spirochaetosis (PIS), respectively. Although some inactivated bacterin and recombinant vaccines have been explored as prophylactic treatments against these species, no effective vaccine is yet available. Immunoproteomics approaches hold the potential for the identification of new, suitable candidates for subunit vaccines against SD and PIS. These strategies take into account the gene products actually expressed and present in the cells, and thus susceptible of being targets of immune recognition. In this context, we have analyzed the immunogenic pattern of two B. pilosicoli porcine isolates (the Spanish farm isolate OLA9 and the commercial P43/6/78 strain) and one B. hyodysenteriae isolate (the Spanish farm V1). The proteins from the Brachyspira lysates were fractionated by preparative isoelectric focusing, and the fractions were analyzed by Western blot with hyperimmune sera from challenged pigs. Of the 28 challenge-specific immunoreactive bands detected, 21 were identified as single proteins by MS, while the other 7 were shown to contain several major proteins. None of these proteins were detected in the control immunoreactive bands. The proteins identified included 11 from B. hyodysenteriae and 28 from the two B. pilosicoli strains. Eight proteins were common to the B. pilosicoli strains (i.e., elongation factor G, aspartyl-tRNA synthase, biotin lipoyl, TmpB outer membrane protein, flagellar protein FlaA, enolase, PEPCK, and VspD), and enolase and PEPCK were common to both species. Many of the identified proteins were flagellar proteins or predicted to be located on the cell surface and some of them had been previously described as antigenic or as bacterial virulence factors. Here we report on the identification and semiquantitative data of these immunoreactive proteins which constitute a unique antigen collection from these bacteria.

A PCR protocol to establish standards for routine mycoplasma testing that by design detects over ninety percent of all known mycoplasma species

“…These non-mycoplasma matches are species of the genera Brachyspira , Micrococcoides , Nitriliruptor , Oligella , and Peptococcus . Because the species Brachyspira hyodysenteriae , Peptococcus niger and Peptococcus simiae are anaerobic, 32 , 33 , 34 they are absent from standard tissue cultures. The remaining cross-reactive aerobic species are the Gram-positive Micrococcoides hystricis and Nitriliruptor alkaliphilus and the Gram-negative Oligella ureolytica , 35 , 36 , 37 all of which have not been reported as cell culture contaminants.…”

Evidence of homologous recombination as a driver of diversity in Brachyspira pilosicoli

“…Furthermore, our study generated an indexed, clustered database of 207 putative virulence factors based on proteins identified in previous studies [60,61]. On average, the B. pilosicoli isolates harboured 196 matches or putative allele assignments.…”

“…A list of 231 putative virulence factors was created by combining and filtering replicate UniProt accession numbers from (i) a list of proteins with pathogenic and virulence potential [ 60 ] and (ii) a list of 26 mostly surface-exposed proteins identified as cross-reacting bands using immune pig sera [ 61 ]. The UniProt accessions were converted into coding sequence (CDS) nucleotide accessions from UniProt ( https://www.uniprot.org/ ) and then downloaded using the batch entrez service ( https://www.ncbi.nlm.nih.gov/sites/batchentrez ).…”

Surface Immunoproteomics Reveals Potential Biomarkers in Alicyclobacillus acidoterrestris

“…This protein is mainly involved in chain elongation during polypeptide synthesis which could bind and transport of aminoacyl-tRNA to the ribosome and interact with GTP in the catalysis of GTP into GDP (Caldas et al, 1998). It is widely distributed in bacteria and has been identified as immunogen in many microorganisms (Fowsantear et al, 2014; Casas et al, 2017).…”