Since pEO5 and pHly152 differ in their origin, size and conjugati

Since pEO5 and pHly152 differ in their origin, size and conjugative transfer, we investigated if STA-9090 cost plasmid α-hly operons have a common origin and evolved independently of chromosomal α-hlyCABD genes in E. coli. In order to explore the genetic relationship between plasmid α-hly genes we investigated five α-hly plasmids originating from canine ETEC strains and four plasmids of porcine ETEC and STEC strains (Table 1). α-hemolysin plasmids were detected by DNA-hybridization of Southern blotted plasmid DNA as described in Material and Methods

(Fig. 1). The size of α-hly plasmids from dogs, pigs, mouse, cattle and human origin varied between 48 kb to 157 kb and other than pEO13, pEO14 and pEO860 all other plasmids were found transferable by conjugation (Table 1). Plasmid profile analysis has shown that the α-hly-plasmids are frequently found together with other large this website plasmids (Fig. 1). Table 1 Relevant properties of strains carrying plasmid and chromosomally encoded α-hly determinants           PCR products with primers pairsa strain Serotype b Origin, reference d hly -plasmid BAY 80-6946 in vivo (kb) Plasmid group 1f/r (678 bp) 32f/r (671 bp) 44f/r (685 bp) 99f/r (650 bp) 72f/r (695 bp) 81f/r (773 bp) C4115 O26:[H11] human, EPEC [21] pEO5 (157) 1 + + + + – - TPE422 Or:H48 E. coli K12 (pEO5) [21] pEO5 (157) 1 +

+ + + – - CB9866 O26:[H11] cattle, EPEC [21] pEO5 (157) 1 + + + + – - CB1027 O26:[H11] human, EPEC [21] pEO5 (157) 1 + + + + – - CB1030 O26:[H11] human, EPEC [21] pEO5 (157) 1 + + + +

– - IP187 O26:[H11] human, EPEC [21] pEO5 (157) 1 + + + + – - 84/2195 Ont:H10 dog [10] pEO9 (146) 1 + + + + – - 84-R O121:H46 dog [10] pEO13 (97) 1 + + + + – - 374 Nintedanib (BIBF 1120) Or:H48 mouse [24] pHly152 (48) 2 + e) + + – - 84-3208 O42:H37 dog, ETEC[10] pEO11 (48) 2 + e) + + – - 84-2573 O70:NM dog, ETEC [10] pEO12 (48) 2 + e) + + – - CB853 O138:H14 pig, STEC [29] pEO853 (145) 3 + f) g) + – - CB855 O138:NM pig, STEC [29] pEO855 (140) 3 + f) g) + – - CB857 O157:NM pig, ETEC [42] pEO857 (97) 3 + f) g) + – - CB860 O149:H10 pig, ETEC [42] pEO860 (48) single + + g) + – - 84-2S O75:H2 dog [10] pEO14 (97) single – - – - – - 536h O6:K15:H31 human UPEC [20] – n.a – - – - + + 536-14 O6:K15:H31 PAI I deletion mutant of 536 [20] – n.a – - – - + – 695/83 O126:H27 human [19] – n.a – - – - – i) J96h O4:K6 human UPEC [46] – n.a – - – - + j) KK6-16 E. cloacae human [26] – n.a k) – - – - – a) primer pairs and size of the PCR products obtained with strains TPE422 (pEO5) (primers 1f/r, 32f/r and 44f/r) and 536 (primers 81f/r and 72f/r) (see Table 2). + = a PCR product of the same size as obtained with strains TPE422 (pEO5) or 536, respectively. – = no PCR product obtained PCR products with other sizes than obtained with the reference strains are indicated for their length in bp.

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