Non-wild Type Research Articles

Molecular epidemiology and azole resistance mechanism study of Candida guilliermondii from a Chinese surveillance system

We studied the molecular epidemiology and mechanism of azole resistance of 164 C. guilliermondii isolates from a nationwide multi-center surveillance program. The isolates were identified by ITS gene sequencing, and the in vitro susceptibility to fluconazole and voriconazole was determined by broth microdilution method. The 14-α-demethylase gene ERG11 was amplified and sequenced, and microsatellite analysis was performed to study the genetic relatedness of the isolates. Amongst the 164 C. guilliermondii isolates, 15 (9.1%) and 17 (10.4%) isolates were assigned to be non-wild type (non-WT) to fluconazole and voriconazole, respectively. Sixteen sequence types (STs) were detected by comparing the amino acid sequence polymorphisms of the ERG11 gene. Fifteen isolates of STs 9, 10, 12, 13, 14, 15 and 16, were all assigned to be non-WT to fluconazole and voriconazole. By microsatellite analysis, 40 different genotypes were identified. Thirty-seven isolates from one hospital (Z1) shared the same ERG11 sequence type (ST 2), microsatellite genotype (PU40) and drug resistance pattern. In conclusion, this is the first molecular epidemiology study of C. guilliermondii in China. The rate of non-WT isolates to azoles was high and the accurate contribution of ERG11 gene mutations to azole resistance need be confirmed by further studies.

Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing

Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods.

Antimicrobial resistance surveillance in Escherichia coli by using normalized resistance interpretation

ObjectivesTo improve antimicrobial surveillance accuracy for results obtained by disk diffusion for porcine Escherichia coli, by comparing traditional clinical breakpoint interpretation with the Normalized Resistance Interpretation (NRI) method. MethodsThe susceptibilities of 921 E. coli isolates from clinically healthy pigs at slaughter age was determined for 15 antimicrobials by the Kirby Bauer disk diffusion technique. NRI with previously established optimal controlled parameters for E. coli ATCC25922 was used to reconstruct the fully susceptible population of the tested E. coli isolates. Based on a lower limit for susceptibility, set at 2.5 standard deviations below the mean of the reconstructed susceptible population, the non-wild type percentage isolates was compared with the clinical resistance percentage. ResultsThe NRI method was applicable for 11 out of the 15 antimicrobials tested.Antimicrobials for which no normal distribution of inhibition zones for the population of susceptible isolates was seen, could not be used to reconstruct the susceptible population. Clinical breakpoints much lower than the epidemiological cut-off values resulted into presumptively identifying isolates as clinically susceptible, but likely carrying acquired resistance determinants. Otherwise, clinical breakpoints did cut through the WT population for several antibiotics tested, categorizing isolates from the WT population as not susceptible. ConclusionsNRI was shown to be a valid method to define the WT population for disk diffusion outcomes, provided a normal distribution of the susceptible bacterial species population is present. Until international harmonization of breakpoints is achieved, it might give rise to a wide application in monitoring antimicrobial resistance in veterinary medicine.

Provisional epidemiological cutoff values for standard broth microdilution susceptibility testing of Flavobacterium columnare.

The gliding aquatic bacterium Flavobacterium columnare causes columnaris disease, a common problem for wild and farmed freshwater fish worldwide. Recently, a broth microdilution method was standardized to test the susceptibility of F. columnare against antimicrobials commonly used in aquaculture. We used this new method to measure the minimal inhibitory concentrations (MICs) of ten antimicrobials against 120 F. columnare isolates. The resulting MIC frequency distributions for each antimicrobial (1 MIC/isolate) were used to estimate epidemiological cut-off values (ECVs) which separate isolates with typical wild-type (WT) susceptibility from isolates with decreased non-wild-type (NWT) susceptibility. We identified 22 NWT isolates with elevated MICs relative to the ECV that covered 99.9% of the MIC distribution against one or more of the antimicrobials: ampicillin, enrofloxacin, erythromycin, florfenicol, flumequine, oxolinic acid or oxytetracycline. Ten of the NWT isolates had decreased susceptibility to a single antimicrobial class, six isolates to two antimicrobial classes and six isolates to three or more antimicrobial classes. The MIC frequency distributions and provisional cut-off values provide data needed to set epidemiological cut-off values to monitor for the development of antimicrobial resistance among F. columnare.

Antimicrobial resistances and virulence markers in Methicillin-resistant Staphylococcus aureus from broiler and turkey: A molecular view from farm to fork

Little is known about the characteristics of MRSA occurring along the broiler and turkey production chains. The aim of this study was to characterise and compare MRSA of turkey and broiler origin sampled on different production levels using a DNA microarray and antimicrobial susceptibility testing. Main differences could be observed in the prevalence of the resistance genes erm(C), aacA-aphD and tet(K) and the number of non-wild type strains with minimum inhibitory concentration values (MICs) above the epidemiological cut-off values (ECOFFs) for gentamicin and kanamycin. Overall, MRSA with non-wild type phenotype for the macrolide-lincosamide-streptogramin group, tetracycline, and trimethoprim were found in more than 70% of poultry isolates. Non-wild type isolates carrying the qacC gene conferring resistance to quaternary ammonium compound disinfectants were found at all production stages in similar frequencies. Regarding the presence of enterotoxin genes in poultry-derived MRSA the enterotoxin gene cluster (egc) was only found in Non-CC (clonal complex) 398 strains. Three CC398 strains harboured the genes sed (from turkey at slaughter and broiler meat) and sea-N315 (from a chicken carcass). One Non-CC398 strain isolated from turkey meat was found positive for the seb gene and also enterotoxin production. Similarity analysis based on selected resistance and virulence markers revealed a high clonality among Non-CC398 isolates. Isolates of the same clonal complex clustered together according to their common virulence and resistance profiles. Strains of CC9 were grouped in two subclusters due to different resistance genes. Our results underline, that there are other poultry associated clones of MRSA (mainly CC9 and CC5) besides the predominant CC398 which are similar in both poultry species.

Host-induced aneuploidy and phenotypic diversification in the Sudden Oak Death pathogen Phytophthora ramorum.

BackgroundAneuploidy can result in significant phenotypic changes, which can sometimes be selectively advantageous. For example, aneuploidy confers resistance to antifungal drugs in human pathogenic fungi. Aneuploidy has also been observed in invasive fungal and oomycete plant pathogens in the field. Environments conducive to the generation of aneuploids, the underlying genetic mechanisms, and the contribution of aneuploidy to invasiveness are underexplored. We studied phenotypic diversification and associated genome changes in Phytophthora ramorum, a highly destructive oomycete pathogen with a wide host-range that causes Sudden Oak Death in western North America and Sudden Larch Death in the UK. Introduced populations of the pathogen are exclusively clonal. In California, oak (Quercus spp.) isolates obtained from trunk cankers frequently exhibit host-dependent, atypical phenotypes called non-wild type (nwt), apparently without any host-associated population differentiation. Based on a large survey of genotypes from different hosts, we previously hypothesized that the environment in oak cankers may be responsible for the observed phenotypic diversification in P. ramorum.ResultsWe show that both normal wild type (wt) and nwt phenotypes were obtained when wt P. ramorum isolates from the foliar host California bay (Umbellularia californica) were re-isolated from cankers of artificially-inoculated canyon live oak (Q. chrysolepis). We also found comparable nwt phenotypes in P. ramorum isolates from a bark canker of Lawson cypress (Chamaecyparis lawsoniana) in the UK; previously nwt was not known to occur in this pathogen population. High-throughput sequencing-based analyses identified major genomic alterations including partial aneuploidy and copy-neutral loss of heterozygosity predominantly in nwt isolates. Chromosomal breakpoints were located at or near transposons.ConclusionThis work demonstrates that major genome alterations of a pathogen can be induced by its host species. This is an undocumented type of plant-microbe interaction, and its contribution to pathogen evolution is yet to be investigated, but one of the potential collateral effects of nwt phenotypes may be host survival.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2717-z) contains supplementary material, which is available to authorized users.

The role of epidemiological cutoff values (ECVs/ECOFFs) in antifungal susceptibility testing and interpretation for uncommon yeasts and moulds

The role of antimicrobial susceptibility testing is to aid in selecting the best agent for the treatment of bacterial and fungal diseases. This has been best achieved by the setting of breakpoints by Clinical Laboratory Standards Institute (CLSI) for prevalent Candida spp. versus anidulafungin, caspofungin, micafungin, fluconazole, and voriconazole. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) also has set breakpoints for prevalent and common Candida and Aspergillus species versus amphotericin B, itraconazole, and posaconazole. Recently, another interpretive category, the epidemiological cut off value, could aid in the early identification of strains with acquired resistance mechanisms. CLSI has postulated that epidemiological cut off values may, with due caution, aid physicians in managing mycosis by species where breakpoints are not available. This review provides (1) the criteria and statistical approach to establishing and estimating epidemiological cut off values (ECVs), (2) the role of the epidemiological cut off value in establishing breakpoints, (3) the potential role of epidemiological cut off values in clinical practice, (4) and the wide range of CLSI-based epidemiological cut off values reported in the literature as well as EUCAST and Sensititre Yeast One-ECVs. Additionally, we provide MIC/MEC (minimal inhibitory concentrations/minimum effective concentrations) ranges/modes of each pooled distribution used for epidemiological cut off value calculation. We focus on the epidemiological cut off value, the new interpretive endpoint that will identify the non-wild type strains (defined as potentially harboring resistance mechanisms). However, we emphasize that epidemiological cut off values will not categorize a fungal isolate as susceptible or resistant as breakpoints do, because the former do not account for the pharmacology of the antifungal agent or the findings from clinical outcome studies.

Variation in the prion protein sequence in Dutch goat breeds.

Scrapie is a neurodegenerative disease occurring in goats and sheep. Several haplotypes of the prion protein increase resistance to scrapie infection and may be used in selective breeding to help eradicate scrapie. In this study, frequencies of the allelic variants of the PrP gene are determined for six goat breeds in the Netherlands. Overall frequencies in Dutch goats were determined from 768 brain tissue samples in 2005, 766 in 2008 and 300 in 2012, derived from random sampling for the national scrapie surveillance without knowledge of the breed. Breed specific frequencies were determined in the winter 2013/2014 by sampling 300 breeding animals from the main breeders of the different breeds. Detailed analysis of the scrapie-resistant K222 haplotype was carried out in 2014 for 220 Dutch Toggenburger goats and in 2015 for 942 goats from the Saanen derived White Goat breed. Nine haplotypes were identified in the Dutch breeds. Frequencies for non-wild type haplotypes were generally low. Exception was the K222 haplotype in the Dutch Toggenburger (29%) and the S146 haplotype in the Nubian and Boer breeds (respectively 7 and 31%). The frequency of the K222 haplotype in the Toggenburger was higher than for any other breed reported in literature, while for the White Goat breed it was with 3.1% similar to frequencies of other Saanen or Saanen derived breeds. Further evidence was found for the existence of two M142 haplotypes, M142 /S240 and M142 /P240 . Breeds vary in haplotype frequencies but frequencies of resistant genotypes are generally low and consequently selective breeding for scrapie resistance can only be slow but will benefit from animals identified in this study. The unexpectedly high frequency of the K222 haplotype in the Dutch Toggenburger underlines the need for conservation of rare breeds in order to conserve genetic diversity rare or absent in other breeds.

Characterization of quinolone resistance mechanisms in Enterobacteriaceae recovered from diseased companion animals in Europe

ComPath is a European monitoring programme dedicated to the collection of bacterial pathogens from diseased dogs and cats to determine their antibiotic susceptibility. The objective was to characterize genetic determinants associated with quinolone resistance among 69 enrofloxacin non-wild type strains selected among 604 non-duplicate Enterobacteriaceae isolates collected in 10EU countries from 2008 to 2010: quinolone resistance determining region (QRDR) and plasmid-mediated quinolone resistance (PMQR). Among them, 17% (12/69) carried at least one PMQR (9/12 qnrB, qnrS or qnrD and 4/12 aac(6′)-Ib-cr) and 83% (57/69) no PMQR. All the Klebsiella pneumoniae isolates chromosomally carried oqxAB . No qepA genes were detected. Eight strains did not carry any mutations in QRDR (4 PMQR-positive and 4 PMQR-negative strains). From the 12 PMQR-positive strains, 4 showed enrofloxacin MICs≤2μg/mL, and 8 MICs≥8μg/mL (resistant). These latter strains carried 1–5 mutations in QRDR, including a ParE I529L mutation. qnrD was found in 2 Proteus mirabilis and the plasmids were similar to pDIJ09-518a previously described. For the 57 non-PMQR strains, 29 strains showed MICs≤2μg/mL (4 with no QRDR mutations, 21 with 1 mutation in GyrA, 4 with 2 mutations in GyrA) and 28 showed enrofloxacin MICs≥8μg/mL carrying at least 2 mutations in QRDR, including a ParE I529L mutation for 2 Escherichia coli strains with a total of 5 QRDR mutations. No GyrB mutations were found. qnr was the major PMQR and qnrD was only detected in Proteus spp. Twelve strains carried at least 4 mutations.

Antimicrobial resistance and population structure of Staphylococcus aureus recovered from pigs farms

Staphylococcus aureus is a burden in human and veterinary medicine. During the last decade, an increasing number of studies reported the presence of livestock-associated methicillin-resistant S. aureus (LA-MRSA) clonal complex (CC) 398 in pigs. During 2013, a survey was performed in pig farms (n=328) randomly selected over Belgium, to monitor the current epidemiological situation of LA-MRSA among asymptomatic pigs and compare with former data to determine possible evolutions. Per farm, nose swabs were taken from 20 animals and pooled. MRSA was detected in 215 farms. Most isolates belonged to CC398 (n=211), and the remaining were ST9/t337 (n=1), ST80/t044 (n=2) and ST239/t4150 (n=1). A large diversity (n=19) of spa-types was found in the CC398 isolates. More than 90% of the isolates were non-wild type (NWT) to tetracycline and trimethoprim. NWT isolates were also found for ciprofloxacin (61.1%), clindamycin (64.4%), erythromycin (57.8%), kanamycin (43.1%) and gentamicin (45.5%). Microarray analysis showed that most CC398 isolates carried genes encoding resistance to tetracycline [tet(M)], macrolide–lincosamide–streptogramin group [erm(B), erm(C), lnu(A), vga(A)], aminoglycosides (aacA-aphD,aa dD, aphA3, sat) and/or phenicols (fexA). One CC398 isolate carried the multi-resistance gene cfr. The non-CC398 isolates carried virulence genes, as the egc-like cluster. The ST80 strain carried the Panton–Valentine leukocidin gene and corresponded to the community-acquired (CA-)MRSA ST80-IV European clone. The MRSA prevalence among pigs in Belgium remains similar to previous studies but a larger diversity in spa-types has been detected in this study. The recovery of CA-MRSA from livestock indicates that one should remain vigilant to the evolution of LA-MRSA in pigs.

Study of wild type and non-wild type distribution pattern of clinical isolates of non-dermatophyte filamentous fungi in ocular specimens based on epidemiological cutoff values of voriconazole, itraconazole & amphotericin-B by disc diffusion method in tertiary care eye institute of Gujarat, India

Study of wild type and non-wild type distribution pattern of clinical isolates of non-dermatophyte filamentous fungi in ocular specimens based on epidemiological cutoff values of voriconazole, itraconazole & amphotericin-B by disc diffusion method in tertiary care eye institute of Gujarat, India

HYBRID AND SELFED SEEDLING PROGENIES OF VITIS HYBRID 'ROGER'S RED' GRAPE SEGREGATE FOR TENDRIL DISTRIBUTION

Grapevines of most species and cultivars produce lateral meristems (tendrils and inflorescences) on at least two successive nodes in three. Lateral meristems complicate rootstock cutting production, since tendrils are removed from rootstock cuttings prior to propagation and since tendrils clinging to wires, canes, or other tendrils make collecting cuttings more time consuming and expensive. To develop improved grapevine rootstocks with reduced lateral meristems, an elite nematode resistant rootstock selection ((Vitis berlandieri × V. riparia) × V. biformis) was crossed with ?Roger?s Red? (Vitis hybrid), and ?Roger?s Red? was also self-pollinated. Seedlings from controlled pollinations were grown in a greenhouse and trained up a stake. Starting at the first node in alternate phyllotaxy (that is, a node with a lateral meristem) and continuing for twelve nodes, the presence or absence of lateral meristems was observed and the type of lateral meristem was reported. There were 141 cross-pollinated seedlings. There were 83 seedlings from ?Roger?s Red? self-pollination. The expected pattern of lateral meristem distribution was wild type: two successive lateral meristem bearing nodes, followed by a single lateral meristem-free node. However, 43 of 83 seedlings from ?Roger?s Red? self-pollination showed two or more successive lateral meristem-free nodes (flanked by lateral meristem-bearing nodes), and 12 of these 43 showed at least two sets of two successive lateral meristem-free nodes. One self-pollinated seedling showed three successive lateral meristem-free nodes in the alternate phyllotaxy. In contrast, in the hybrid population only 33 of 141 seedlings showed the non-wild type pattern of two successive lateral meristem-free nodes, and only 6 of these 33 showed at least two sets of two successive lateral meristem-free nodes. Tendril distribution is under genetic control and the production of extra tendril-free nodes was segregating in these populations. Dominant alleles at two genes are required for the mutant phenotype. A genetic model and the gene names Successive Tendril free Node-1 and Successive Tendril free Node-2 are proposed.

Candida en la cavidad oral de pacientes con VIH en Cali, Colombia: determinación de especies y sensibilidad al fluconazol

Introduction: Candida albicans is the most frequently isolated yeast from the oral cavity of HIV-infected individuals. The use of fluconazole has increased thenumber of resistant or less-sensitive Candida species different from C. albicans, to this antifungal agent. Objective: To establish the Candida species present in the oral cavity of HIV-infected individuals at a hospital in Cali (Colombia), their population densities, and the susceptibility to fluconazole of species different from C. albicans. Materials and methods: Samples were cultured in CHROMagar Candida and the number of colony forming units (CFU) was counted. Yeast identification was done with API 20C Aux, and the susceptibilitytests to fluconazole, by Etest. Results: 230 patients were studied, and 202 isolates were obtained: 106 single and 96 mixed. C. albicans predominated, followed by C. dubliniensis and C.glabrata. Candida species other than C. albicans predominated in counts lower than 400 CFU/mL. Susceptibility study to fluconazole of species different fromC. albicans showed that 14 (40 %) of the isolates were susceptible dose-dependent and 7 (20 %), resistant. Conclusion: In the studied population, the oral cavity was colonized by non-wild type isolates that represent a risk for the development of oropharyngeal candidiasis resistant to fluconazole treatment.

Characterization of methicillin-resistant Staphylococcus sciuri isolates from industrially raised pigs, cattle and broiler chickens.

This study aimed at assessing the epidemiology and genetic diversity of methicillin-resistant Staphylococcus sciuri (MRSS) from different farm animal species. Nasal swabs were collected from 200 pigs, 100 dairy cows, 100 beef cows, 150 veal calves and 200 broilers. Colonies were isolated on selective media containing cefoxitin and the mecA gene was detected by PCR. Antimicrobial resistance was determined by broth microdilution. The genetic diversity was assessed by PFGE and resistance and virulence genes were detected by microarray analysis. The total MRSS prevalence at the animal level was estimated at 9.5%, varying from ∼10% in veal (13.3%), broilers (12.5%) and dairy cows (10.0%) to 6.5% in pigs and 3.0% in beef cows. mecA was detected in all isolates. SCCmec elements of type III and non-typeable ones were seen most frequently. More than 90% of isolates were non-wild-type (NWT) for gentamicin, penicillin, tiamulin, clindamycin and quinupristin/dalfopristin. The frequency of NWT isolates for fusidic acid and trimethoprim ranged between 78% and 87%. PFGE analysis allowed distinction between two major clusters. Most isolates tested by microarray carried erm and tet genes. Virulence genes were also detected, including an isa gene encoding an immune-evasion factor and the hsdS2 gene encoding a site-specific deoxyribonuclease. This study shows that multiresistant MRSS is carried by different farm animal species. Although some animals shared the same strain, PFGE showed different patterns, indicating high diversity among the MRSS isolates recovered. The absence of clusters associated with a certain animal species suggests low host specificity.

Antifungal susceptibilities of Candida isolates causing bloodstream infections at a medical center in Taiwan, 2009-2010.

We used the Sensititre YeastOne (SYO) method (Trek Diagnostic Systems) to determine the MICs of nine antifungal agents against 474 nonduplicate blood Candida isolates. The MIC results were interpreted according to updated clinical breakpoints (CBPs) recommended by the Clinical and Laboratory Standards Institute (CLSI; document M27-S4) or epidemiology cutoff values (ECVs). The rates of fluconazole susceptibility were 99.2% (234/236) in Candida albicans, 86.7% (85/98) in C. tropicalis, and 97.7% (42/43) in C. parapsilosis. Among the 77 isolates of C. glabrata, 90.9% showed dose-dependent susceptibility (S-DD) to fluconazole. Nearly all isolates of C. albicans, C. parapsilosis, and C. krusei were susceptible to voriconazole; however, rates of voriconazole susceptibility were 78.6% in C. tropicalis. Few isolates of C. albicans (n = 5; 2.1%) and C. glabrata (n = 3; 3.9%), no isolates of C. parapsilosis, C. krusei, and C. guilliermondii, but 62.2% (n = 51) of C. tropicalis isolates were non-wild type for posaconazole susceptibility. For itraconazole susceptibility, 98.3% of C. albicans isolates were wild type, and 3.9% (n = 3) of C. glabrata isolates were non-wild type. Almost all of the isolates tested (>97% for all species) were susceptible to both micafungin and anidulafungin. All isolates tested were found to be wild type for amphotericin B susceptibility, with MICs of <1 μg/ml. Further evaluation is needed to establish CBPs of antifungal agents by the 24-h SYO method for the management of patients with candidemia or other invasive candida infections.

Relationship between UDP-glucuronosyltransferase 1A1 gene polymorphisms and toxicity in patients with cancer treated with irinotecan-based chemotherapy

Objective To investigate the incidence and severity of adverse events in pateints with solid tumor treated with irinotecan-based chemotherapy,and their relationship with UDP-glucuronosyltransferase 1A1 (UGT1A1) gene polymorphisms.Methods Sixty-seven tumor patients treated with irinotecan-based chemotherapy were enrolled.Genomic DNA was extracted from peripheral blood,and the UGT1 A1 * 28 and UGT1 A1 * 6 genotypes were determined by direct sequencing.The adverse events during therapy were observed.The relationship between UGT1A 1 gene polymorphisms and toxicity of irinotecan was analyzed.Results The distribution of the genotypes in 67 cancer patients was as follows:UGT1 A1 * 28 wild type genotype TA6/6 (51,76.1％),heterozygous genotype TA6/7 (12,17.8％),and homozygous genotype TA7/7 (4,6.0％); UGT1A1 * 6 wild type genotype G/G (43,64.2％),heterozygous genotype G/A (18,26.9％),and homozygous genotype A/A (6,9.0％).The incidence of grade 3 and 4 diarrhea and thrombocytopenia in the patients with UGT1A1 * 28 (TA6/7 and TA7/7) was 56.3％ and 31.3％ respectively,significantly higher than 25.5％ and 13.7％ in the patient with wild type UGT1A1 * 28 (TA6/6).The incidence of grade 3-4 diarrhea in the patients with UGT1A1 * 6 (G/A + A/A) was 50.0％,significantly higher than 1 1.6％ in the patient with wild type UGT1 A1 * 6 (G/G).Toxicity rate in patients with the non-wild type UGT1 A1 gene polymorphisms was significantly higher than that in the patients with wild type UGT1A1 gene (P＜0.05).Conclusion The UGT1A1 * 28 (TA6/7 and TA7/7) genotypes increase the risk of grade 3-4 diarrhea or thrombocytopenia,and the UGT1A1 * 6 (G/A and A/A) genotypes increase the risk of grade 3-4 diarrhea in tumor patients treated with irinotecan-based chemotherapy. Key words: UDP-glucuronosyltransferase 1A1 ; Gene polymorphisms; Irinotecan

Effect of herbs on preventing diarrhea caused by irinotecan and its correlation with gene polymorphism of UGT1A1*28

Objective: This study aimed to determine the function of herbs in preventing diarrhea after irinotecan chemotherapy and analyze the efficacy of the herbs based on UGT1A1*28 gene polymorphism. Methods: A total of 200 patients admitted to the Department of Synergistic Chinese and Western Medicine, Tianjin Medical University Cancer Institute and Hospital between October 2011 and May 2013 were randomly divided into the control (chemotherapy alone) and herb (chemotherapy combined with herbs) groups. All patients consented to UGT1A1*28 gene polymorphism detection prior to chemotherapy. Herbs were administered from 2 d prior to chemotherapy to 5 d post chemotherapy, with or without the regimen of fluorouracil, folinic acid, and irinotecan. Adverse reactions were recorded, and short-term effect was evaluated regularly. Results: A total of 144 patients had TA6/6 wild genotype, and another 56 patients had non-wild genotype (12 of the 56 cases were TA7/7 homozygous, and the other 44 cases were TA6/7 hybrid). A total of 58 patients experienced grades 2 to 4 diarrhea. A 14% decrease in the incidence of diarrhea was observed in the herb group compared with that of the control group (22% vs. 36%, P=0.029). In addition to diarrhea, grades 2 to 4 vomiting was significantly lower in the herb group than in the control group (15% vs. 27%, P=0.037). The overall response rate was 37.5%. No significant difference was found between the two groups (40% vs. 35%, P=0.465). The incidences of grades 2 to 4 diarrhea (22.9% vs. 44.6%, P=0.002) and grades 2 to 4 vomiting (23.2% vs. 16.7%, P=0.016) were lower in patients with the UGT1A1*28 wild genotype than in those with the non-wild genotype. However, in the herb group, the incidences of grades 2 to 4 diarrhea (22.2% vs. 21.9%, P=0.974) and vomiting (18.5% vs. 13.7%, P=0.777) were not significant between the non-wildand wild-type groups. Conclusion: Herbs can effectively pre作者单位:1天津医科大学肿瘤医院中西医结合科,国家肿瘤临床医学研究中心,天津市肿瘤防治重点实验室(天津市300060);2消化肿瘤内科; 3结直肠肿瘤科;4生化及分子生物学研究室 􀆽本文课题受天津市中医药管理局中医中西医结合科研项目(编号:11107)资助 通信作者:谢广茹 xieguangru@126.com 1441 中国肿瘤临床 2013年第40卷第23期 Chin J Clin Oncol 2013, Vol. 40, No. 23 www.cjco.cn vent the late diarrhea caused by irinotecan, which is also applicable in UGT1A1*28 non-wild genotype patients. Incidence of diarrhea was obviously higher in the cases with UGT1A1*28 non-wild type than in those with wild genotype. Hence, the UGT1A1*28 gene type should be detected prior to chemotherapy with irinotecan.

Genetic lineages of Salmonella enterica serovar Kentucky spreading in pet reptiles

The purpose of the study was to define genetic diversity of reptilian Salmonella enterica serovar (S.) Kentucky isolates and their epidemiological relations to the ones from poultry, food, and environmental origin in Poland. Between 2010 and 2012 twenty-four S. Kentucky isolates derived from snakes (N=8), geckos (N=7), chameleons (N=4), agamas (N=1), lizard (N=1), and environmental swabs taken from reptile exhibition (N=3) were identified. They were characterized with antimicrobial minimal inhibitory concentration testing, XbaI–PFGE and MLST typing. The profiles compared to S. Kentucky available in BioNumerics local laboratory database (N=40) showed 67.3% of relatedness among reptile isolates. Three genetic lineages were defined. The first lineage gathered 20 reptile isolates with 83.4% of similarity and wild-type MICs for all antimicrobials tested but streptomycin in single case. The remaining three reptilian and one post-exhibition environment S. Kentucky isolates were clustered (87.2%) with isolates originating from poultry, mainly turkey, food, and environment and presented variable non-wild type MICs to numerous antimicrobials. The third S. Kentucky lineage was composed of two isolates from feed (96.3%). The results suggest diverse sources and independent routes of infection. Most of the isolates belonged to reptile-associated clones spread both horizontally and vertically. Simultaneously, PFGE profiles and MLST type indistinguishable from the ones observed in poultry point out carnivore reptiles as possible vector of infection with multidrug and high-level ciprofloxacin resistant (MIC≥8mg/L) S. Kentucky. Public awareness and education are required to prevent potential reptile-associated S. Kentucky infections in humans.

Cross-Sectional Study on Prevalence and Molecular Characteristics of Plasmid Mediated ESBL/AmpC-Producing Escherichia coli Isolated from Veal Calves at Slaughter

ObjectivesThe presence of ESBL/AmpC-producing E. coli in cattle has been reported previously, however information on veal calves is limited. This study describes the prevalence and molecular characteristics of E. coli with non-wild type susceptibility to cefotaxime in veal calves at slaughter.MethodsFaecal samples from 100 herds, 10 individual animals per herd, were screened for E. coli with non-wild type susceptibility for cefotaxime. Molecular characterization of ESBL/AmpC genes and plasmids was performed on one isolate per herd by microarray, PCR and sequence analysis.Results66% of the herds were positive for E. coli with non-wild type susceptibility for cefotaxime. Within-herd prevalence varied from zero to 90%. 83% of E. coli producing ESBL/AmpC carried bla CTX-M genes, of which bla CTX-M-1, bla CTX-M-14 and bla CTX-M-15 were most prevalent. The dominant plasmids were IncI1 and IncF-type plasmids.ConclusionsA relatively high prevalence of various bla CTX-M producing E. coli was found in veal calves at slaughter. The genes were mainly located on IncI1 and IncF plasmids.

Vibrio ordalii antimicrobial susceptibility testing—Modified culture conditions required and laboratory-specific epidemiological cut-off values

The present study aimed to determine oxytetracycline (OTC), florfenicol (FLO) and oxolinic acid (OXO) MICs and zone diameters for 24 Chilean Vibrio ordalii isolates using the methods for broth dilution susceptibility testing of bacteria isolated from aquatic animals and the methods for antimicrobial disk susceptibility testing of bacteria isolated from aquatic animals guidelines published by the Clinical Laboratory Standards Institute (CLSI). The results were then used in a normalized resistance interpretation (NRI) analysis to establish tentative laboratory-specific epidemiological cut-off (ECOFF) values. MIC results were similar at the two tested temperatures (22 °C and 18 °C). At 18 °C, the NRI analysis of OTC, FLO and OXO MIC data calculated laboratory-specific ECOFF values and non-wild-type (NWT) rates to be ≤4 mg/l (24%), ≤16 mg/l (4%) and ≤8 mg/l (25%), respectively. Tests performed with all V. ordalii isolates following the officially recommended incubation temperature (22 °C) revealed difficulties in measuring inhibition zone diameters. When disk diffusion tests were performed using Mueller-Hinton agar with 1% NaCl (MHA-1) at 18 °C the inhibition zone diameter distributions showed the formation of WT populations which could be defined using NRI analysis. For OTC the laboratory-specific ECOFF value was ≥38 mm with NWT rate of 16.7%. For FLO and OXO, the laboratory-specific ECOFF values were ≥38 and ≥40, respectively, generating NWT rates of 25 and 46%, respectively. Although the CLSI suggests testing Vibrio spp. on MHA-1 at 22 °C, we found measurements of the 24 isolates were better defined and normally distributed at 18 °C. This is the first study determining the MIC and disk diffusion test of V. ordalii isolated from diseased salmonids, where laboratory-specific ECOFF values could be established. Also resistance to OTC, FLO and OXO among some Chilean isolates was demonstrated.