Showing papers by "Marilyn C. Roberts published in 2006"

Mutans Streptococci Dose Response to Xylitol Chewing Gum

Abstract: Xylitol is promoted in caries-preventive strategies, yet its effective dose range is unclear This study determined the dose-response of mutans streptococci in plaque and unstimulated saliva to xylitol gum Participants (n = 132) were randomized: controls (G1) (sorbitol/maltitol), or combinations giving xylitol 344 g/day (G2), 688 g/day (G3), or 1032 g/day (G4) Groups chewed 3 pellets/4 times/d Samples were taken at baseline, 5 wks, and 6 mos, and were cultured on modified Mitis Salivarius agar for mutans streptococci and on blood agar for total culturable flora At 5 wks, mutans streptococci levels in plaque were 10x lower than baseline in G3 and G4 (P = 0007/0003) There were no differences in saliva At 6 mos, mutans streptococci in plaque for G3 and G4 remained 10x lower than baseline (P = 0007/004) Saliva for G3 and G4 was lower than baseline by 8 to 9x (P = 0011/0038) Xylitol at 644 g/day and 1032 g/day reduces mutans streptococci in plaque at 5 wks, and in plaque and unstimulated sal

CTX-M-15 extended-spectrum β-lactamase from Nigerian Klebsiella pneumoniae

Abstract: Objectives In this study, extended-spectrum beta-lactamases (ESBLs) were characterized from 30 selected multidrug-resistant Klebsiella pneumoniae strains isolated from patients with community-acquired urinary tract infections from Southwest Nigeria. Methods The beta-lactamases were phenotypically characterized using isoelectric focusing, genotypically characterized using PCR assays and hybridization of the PCR products. Two of the bla(CTX-M) genes were completely sequenced. The location of the CTX-M-type genes was determined using transformation, DNA-DNA hybridization, PCR assays and hybridization of the PCR products from the Escherichia coli transformants. Results All 30 isolates produced at least one beta-lactamase. Seventeen of the isolates were resistant to cefotaxime, and had > or =100-fold reduction in susceptibility with cefotaxime plus clavulanic acid (4 mg/L), indicating the presence of an ESBL. The 17 isolates were shown to have bla(CTX-M) genes that were associated with large plasmids (> or =58 kb), which also carried a tetracycline resistance gene, tet(A), and various aminoglycoside resistance genes. Two CTX-M-type genes were sequenced and had amino acid sequences indistinguishable from previously sequenced CTX-M-15 beta-lactamases. The ISEcp1 element was located upstream of bla(CTX-M-15) in the same position as previously described. In addition, 23 of the isolates produced TEM beta-lactamases, 27 produced SHV beta-lactamases and four produced AmpC beta-lactamases. Conclusions Thirty K. pneumoniae produced multiple beta-lactamases, with 57% producing CTX-M enzymes. This is the first characterization of CTX-M-15-positive K. pneumoniae in Western Africa.

New antibiotic resistance genes associated with CTX-M plasmids from uropathogenic Nigerian Klebsiella pneumoniae.

Abstract: Objectives: To determine antibiotic resistance genes associated with 17 Nigerian CTX-M-positive Klebsiella pneumoniae plasmids from patients with community-acquired urinary tract infections. Methods: The size and restriction patterns of the plasmids were determined, and antibiotic resistance genes were identified using DNA-DNA hybridization, PCR assays, hybridization of PCR products with internal probes, and sequencing. Results: All CTX-M plasmids were large (58-320 kb) and carried the following genes: aac(6')-Ib (aminoglycoside resistance) which included aac(6')-Ib-cr (aminoglycoside-fluoroquinolone resistance), aadA2 (aminoglycoside resistance), erm(B) (macrolide-lincosamide-streptogramin B resistance), bla TEM-1 (ampicillin resistance), tet(A) (tetracycline resistance), sul1 (sulphonamide resistance), dfr (trimethoprim resistance) and intl1, an integrase associated with class 1 integrons. Eleven (65%) plasmids carried an mph(A) gene (macrolide resistance), seven (41%) plasmids carried a qnrB1 gene (low-level quinolone resistance) and four (24%) plasmids carried multiple cat genes (chloramphenicol resistance). catA2, catA3 and qnrB1 genes and a 6 kb PstI fragment, carrying the bla CTX-M gene, were sequenced. Conclusions: This is the first description of catA2 and catA3 genes in Klebsiella spp. and the first description of the erm(B) and floR genes associated with a CTX-M plasmid. This is also the first report of qnrB1 and aac(6')-Ib-cr in isolates from Africa and the first report of these two genes on the same plasmid.

Staphylococcus efflux msr(A) gene characterized in Streptococcus, Enterococcus, Corynebacterium, and Pseudomonas isolates.

Abstract: The staphylococcal msr(A) gene, coding for a macrolide efflux protein, was identified in three new gram-positive genera and one gram-negative genus. These msr(A) genes shared 99 to 100% identity with each other and the staphylococcal gene. This study demonstrates that the msr(A) gene has a wider host range than previously reported.

Antibiotic resistance genes in multidrug-resistant Enterococcus spp. and Streptococcus spp. recovered from the indoor air of a large-scale swine-feeding operation.

Abstract: Aims: In this study, multidrug-resistant bacteria previously recovered from the indoor air of a large-scale swine-feeding operation were tested for the presence of five macrolide, lincosamide and streptogramin (MLS) resistance genes and five tetracycline (tet) resistance genes. Methods and Results: Enterococcus spp. (n ¼ 16) and Streptococcus spp. (n ¼ 16) were analysed using DNA‐DNA hybridization, polymerase chain reaction (PCR) and oligoprobing of PCR products. All isolates carried multiple MLS resistance genes, while 50% of the Enterococcus spp. and 44% of the Streptococcus spp. also carried multiple tet resistance genes. All Enterococcus spp. carried erm(A) and erm(B), 69% carried erm(F), 44% carried mef(A), 75% carried tet(M), 69% carried tet(L) and 19% carried tet(K). All Streptococcus spp. carried erm(B), 94% carried erm(F), 75% carried erm(A), 38% carried mef(A), 50% carried tet(M), 81% carried tet(L) and 13% carried tet(K). Conclusions: Multidrug resistance among airborne bacteria recovered from a swine operation is encoded by multiple MLS and tet resistance genes. These are the first data regarding resistance gene carriage among airborne bacteria from swine-feeding operations. Significance and Impact of the Study: The high prevalence of multiple resistance genes reported here suggests that airborne Gram-positive bacteria from swine operations may be important contributors to environmental reservoirs of resistance genes.

Linear response of mutans streptococci to increasing frequency of xylitol chewing gum use: a randomized controlled trial [ISRCTN43479664]

Abstract: Xylitol is a naturally occurring sugar substitute that has been shown to reduce the level of mutans streptococci in plaque and saliva and to reduce tooth decay. It has been suggested that the degree of reduction is dependent on both the amount and the frequency of xylitol consumption. For xylitol to be successfully and cost-effectively used in public health prevention strategies dosing and frequency guidelines should be established. This study determined the reduction in mutans streptococci levels in plaque and unstimulated saliva to increasing frequency of xylitol gum use at a fixed total daily dose of 10.32 g over five weeks. Participants (n = 132) were randomized to either active groups (10.32 g xylitol/day) or a placebo control (9.828 g sorbitol and 0.7 g maltitol/day). All groups chewed 12 pieces of gum per day. The control group chewed 4 times/day and active groups chewed xylitol gum at a frequency of 2 times/day, 3 times/day, or 4 times/day. The 12 gum pieces were evenly divided into the frequency assigned to each group. Plaque and unstimulated saliva samples were taken at baseline and five-weeks and were cultured on modified Mitis Salivarius agar for mutans streptococci enumeration. There were no significant differences in mutans streptococci level among the groups at baseline. At five-weeks, mutans streptococci levels in plaque and unstimulated saliva showed a linear reduction with increasing frequency of xylitol chewing gum use at the constant daily dose. Although the difference observed for the group that chewed xylitol 2 times/day was consistent with the linear model, the difference was not significant. There was a linear reduction in mutans streptococci levels in plaque and saliva with increasing frequency of xylitol gum use at a constant daily dose. Reduction at a consumption frequency of 2 times per day was small and consistent with the linear-response line but was not statistically significant.

Distribution of Tetracycline Resistance Genes in Actinobacillus pleuropneumoniae Isolates from Spain

Abstract: Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia. Tetracycline is used for therapy of this disease, and A. pleuropneumoniae carrying the tet(B) gene, coding for an efflux protein that reduces the intercellular tetracycline level, has been described previously. Of the 46 tetracycline-resistant (Tc(r)) Spanish A. pleuropneumoniae isolates used in this study, 32 (70%) carried the tet(B) gene, and 30 of these genes were associated with plasmids. Eight (17%) isolates carried the tet(O) gene, two (4%) isolates carried either the tet(H) or the tet(L) gene, and all these genes were associated with plasmids. This is the first description of these tet genes in A. pleuropneumoniae. The last two Tc(r) isolates carried none of the tet genes examined. Except for tet(O)-containing plasmids, the other 34 Tc(r) plasmids were transformable into an Escherichia coli recipient. Two plasmids were completely sequenced. Plasmid p11745, carrying the tet(B) gene, was 5,486 bp and included a rep gene, encoding a replication-related protein, and two open reading frames (ORFs) with homology to mobilization genes of Neisseria gonorrhoeae plasmid pSJ7.4. Plasmid p9555, carrying the tet(L) gene, was 5,672 bp and, based on its G+C content, consisted of two regions, one of putative gram-positive origin containing the tet(L) gene and the other comprising four ORFs organized in an operon-like structure with homology to mobilization genes in other plasmids of gram-negative bacteria.

The presence of a conjugative Gram-positive Tn2009 in Gram-negative commensal bacteria

Abstract: Objectives To determine whether mef(A)-msr(D) and tet(M) genes are linked in representative Gram-negative isolates and/or transferred together during conjugation. To molecularly characterize the Acinetobacter junii element and compare the structure and sequence with the non-conjugative Streptococcus pneumoniae Tn2009 element. Methods PCR assays, DNA-DNA hybridization and sequencing of PCR products were used. Nucleotide sequences were determined at the integration site of the mef(A) element into Tn916 and upstream and downstream flanking regions of the element. Results A total of 10 mef(A)-msr(D)- and tet(M)-positive isolates carried conjugative element(s). The A. junii Tn2009 element was indistinguishable from S. pneumoniae Tn2009. The region upstream of the A. junii Tn2009 contained an orf that was 89-91% identical to an S. pneumoniae spr1206 gene found upstream of the streptococcal Tn2009. In the A. junii, the spr1206 gene was separated by 67 bp from the end of the Tn2009, while 29 bp were found separating spr1206 from the streptococcal Tn2009. The 1201 bp downstream A. junii sequences included 913 unique sequences. Conclusions A total of 10 different Gram-negative genera were found to carry the tet(M) genes, including the first description in three genera (Citrobacter, Proteus and Stenotrophomonas). All isolates were able to transfer the genes into > or =1 recipient with macrolide selection. Over 3000 bp were sequenced on each side of the insertion mef junction region in the A. junii and were indistinguishable from the streptococcal Tn2009. The A. junii Tn2009 element was flanked by an S. pneumoniae gene upstream and a unique sequence downstream, suggesting that the A. junii Tn2009 could be part of a larger element.

Tetracycline Resistant Plasmids from Uropathogenic Escherichia coli from Southwestern Nigeria

Abstract: (2006). Tetracycline Resistant Plasmids from Uropathogenic Escherichia coli from Southwestern Nigeria. Journal of Chemotherapy: Vol. 18, No. 1, pp. 112-114.