The first report of the qnrB19, qnrS1 and aac(6´)-Ib-cr genes in urinary isolates of ciprofloxacin-resistant Escherichia coli in Brazil

In this study, we investigated the presence of plasmid-mediated quinolone resistance (PMQR) genes among 101 ciprofloxacin-resistant urinary Escherichia coli isolates and searched for mutations in the quinolone-resistance-determining regions (QRDRs) of the DNA gyrase and topoisomerase IV genes in PMQR-carrying isolates. Eight isolates harboured the qnr and aac(6')-Ib-cr genes (3 qnrS1, 1 qnrB19 and 4 aac(6')-Ib-cr). A mutational analysis of the QRDRs in qnr and aac(6')-Ib-cr-positive isolates revealed mutations in gyrA, parC and parE that might be associated with high levels of resistance to quinolones. No mutation was detected in gyrB. Rare gyrA, parC and parE mutations were detected outside of the QRDRs. This is the first report of qnrB19, qnrS1 and aac(6')-Ib-cr -carrying E. coli isolates in Brazil.     

Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli: VIII. The structure of bacteriophage φ80d3ilv+su+7, including the mapping of the ribosomal RNA genes

The sequences present on the DNA of the transducing phage, φ80d₃ilv⁺su⁺7 have been mapped by electron microscope heteroduplex methods. In addition to some φ80 sequences, the phage DNA contains sequences from the extreme counterclockwise region and from the extreme clockwise region of the bacterial chromosomal part of F14. The former includes ilv, the latter a 16 S and a 23 S ribosomal RNA gene. These two regions are joined on the transducing phage DNA by the 2.8 to 8.5F sequence.By direct observation of the structure of the rRNA/DNA hybrids, the 16 S and 23 S genes have lengths of 1.38 ± 0.14 and 2.66 ± 0.17 kilobases. They are separated by a spacer of length 0.57 ± 0.13 kilobases.The rRNA genes (rrn) of φ80d₃ilv⁺su⁺7 are derived from and are identical with the rrnB gene set of F14. In heteroduplexes between the rrnB gene set of φ80d₃ilv⁺su⁺7, and the rrnA gene set of F14 we observe that there is a region of non-homology of length 0.25 ± 0.06 kilobases within the spacer sequence. This confirms observations in the preceding paper on the structure of out-of-register duplexes of the two rRNA gene sets of F14.A model for the integration and excision events involved in the formation of φ80d₃ilv⁺su⁺ 7 from φ80dmet(K) is proposed.     

Effect of increasing the dietary tryptophan to lysine ratio on plasma levels of tryptophan,kynurenine and urea and on production traits in weaner pigs experimentally infected with an enterotoxigenic strain of Escherichia coli‡

This experiment examined if immune system stimulation of weaner pigs, initiated by inoculation an enterotoxigenic strain of Escherichia coli (ETEC), increased the requirement for dietary tryptophan (Trp), modulated the inflammatory response, altered plasma levels of Trp and its metabolite kynurenine (Kyn) and effected post-weaning diarrhoea. Individually housed pigs (n = 72) weaned at 21 d of age were allocated to one of six treatments (n = 12) according to a two by three factorial arrangement of (1) with or without ETEC infection and (2) three dietary ratios of standardised ileal digestible (SID) Trp to lysine (Lys) (SID Trp:Lys) of 0.16, 0.20 or 0.24, in a completely randomised block design. Pigs had ad libitum access to diets (per kg 14.13 MJ ME, 12.4 g SID Lys, 195 g crude protein) for 3 weeks after weaning. Pigs were infected with ETEC (O149:K98:K88) at 72, 96 and 120 h after weaning and then bled on day 3, 11 and 19. An increased dietary Trp:Lys ratio increased plasma Trp and Kyn (p < 0.001) without effect of infection. On day 3, pigs fed 0.24 SID Trp:Lys had lower levels of plasma urea than at 0.20 Trp:Lys (p = 0.047) and on day 11, plasma urea was lower at 0.20 than at 0.16 SID Trp:Lys (p = 0.007). Infection increased (p = 0.039) the diarrhoea index and deteriorated faecal consistency from day 4–10 (p < 0.05). Treatments did not affect haptoglobin and acid soluble glycoprotein levels or daily gain and feed intake. However, 0.24 SID Trp:Lys improved (p = 0.021) feed efficiency without an effect of infection. In conclusion, in the absence of dietary antibiotic growth promotants, increasing the dietary SID Trp:Lys ratio to 0.24 improved feed conversion ratio after weaning and increased plasma levels of Trp and Kyn regardless of infection with E. coli.     

Study of the role played by NfsA,NfsB nitroreductase and NemA flavin reductase from <Emphasis Type="Italic">Escherichia coli</Emphasis> in the conversion of ethyl 2-(2′-nitrophenoxy)acetate to 4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one (D-DIBOA), a benzohydroxamic acid with interesting biological properties

Benzohydroxamic acids, such as 4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one (D-DIBOA), exhibit interesting herbicidal, fungicidal and bactericidal properties. Recently, the chemical synthesis of D-DIBOA has been simplified to only two steps. In a previous paper, we demonstrated that the second step could be replaced by a biotransformation using Escherichia coli to reduce the nitro group of the precursor, ethyl 2-(2′-nitrophenoxy)acetate and obtain D-DIBOA. The NfsA and NfsB nitroreductases and the NemA xenobiotic reductase of E. coli have the capacity to reduce one or two nitro groups from a wide variety of nitroaromatic compounds, which are similar to the precursor. By this reason, we hypothesised that these three enzymes could be involved in this biotransformation. We have analysed the biotransformation yield (BY) of mutant strains in which one, two or three of these genes were knocked out, showing that only in the double nfsA/nfsB and in the triple nfsA/nfsB/nemA mutants, the BY was 0%. These results suggested that NfsA and NfsB are responsible for the biotransformation in the tested conditions. To confirm this, the nfsA and nfsB open reading frames were cloned into the pBAD expression vector and transformed into the nfsA and nfsB single mutants, respectively. In both cases, the biotransformation capacity of the strains was recovered (6.09 ± 0.06% as in the wild-type strain) and incremented considerably when NfsA and NfsB were overexpressed (40.33% ± 9.42% and 59.68% ± 2.0% respectively).