Pyelonephritis in Japan caused by Salmonella enterica subspecies arizonae.

Salmonella enterica subsp. arizonae is a bacteria commonly found in the gut of reptiles. In humans, infections caused by this organism are rare. Most cases originate from southwestern United States, where rattlesnake products are often used in traditional medicine. In Asia, only a few cases have been described. This case report documents a case involving a 64-year-old woman with pyelonephritis caused by S. arizonae in Japan. She had no history of contact with reptiles or foreign travel. The likely route of transmission is unclear. She was treated with cephalosporins for 14 days and the pyelonephritis appeared to be resolved; however recurrence occurred twice -after two weeks and then after one month. Prolonged antibiotic therapy with amoxicillin resolved the infection. This case demonstrates that pyelonephritis associated with S. arizonae can be found outside of the typical geographic region and may not be associated with typical animal hosts.

Antimicrobial activity of leucine-substituted decoralin analogs with lower hemolytic activity.

Linear cationic α-helical antimicrobial peptides are promising chemotherapeutics. Most of them act by different mechanisms, making it difficult to microorganisms acquiring resistance. Decoralin is an example of antimicrobial peptide; it was described by Konno et al. and presented activity against microorganisms, but with pronounced hemolytic activity. We synthesized leucine-substituted decoralin analogs designed based on important physicochemical properties, which depend on the maintenance of the amphiphilic α-helical tendency of the native molecule. Peptides were synthesized, purified, and characterized, and the conformational studies were performed. The results indicated that the analogs presented both higher therapeutic indexes, but with antagonistic behavior. While [Leu]10 -Dec-NH2 analog showed similar activity against different microorganisms (c.a. 0.4-0.8 µmol L-1 ), helical structuration, and some hemolytic activity, [Leu]8 -Dec-NH2 analog did not tend to helical structure and presented antimicrobial activities two orders higher than the other two peptides analyzed. On the other hand, this analog showed to be the less hemolytic (MHC value = 50.0 µmol L-1 ). This approach provided insight for understanding the effects of the leucine substitution in the amphiphilic balance. They led to changes on the conformational tendency, which showed to be important for the mechanism of action and affecting antimicrobial and hemolytic activities. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Enantioselective syntheses of both enantiomers of 9'-dehydroxyimperanene and 7,8-dihydro-9'-dehydroxyimperanene and the comparison of biological activity between 9-norlignans and dihydroguaiaretic acids.

To estimate the effect of methyl group of dihydroguaiaretic acid, which shows many kinds of biological activities, on biological activity, both enantiomers of 9'-dehydroxyimperanene (5, 6) and 7,8-dihydro-9'-dehydroxyimperanene (7, 8) lacking one of the methyl groups of dihydroguaiaretic acid were synthesized. (S)-7,8-Dihydro-9'-dehydroxyimperanene (7) showed 4-6-fold higher cytotoxic activity than all stereoisomers of dihydroguaiaretic acid (2-4). The IC50 values of (S)-7,8-dihydro-9'-dehydroxyimperanene (7) against HL-60 and HeLa cells were 6.1µM and 5.6µM, respectively. Though only one of three stereoisomers of dihydroguaiaretic acid showed antibacterial activity against a gram negative bacterium, both enantiomers of 5-8 showed antibacterial activity against a gram negative bacterium. This is a Letter on biological activity of 9-norlignan, in which one of methyl groups of lignan is absent.

Lipase-catalyzed synthesis of palmitanilide: Kinetic model and antimicrobial activity study.

Enzymatic syntheses of fatty acid anilides are important owing to their wide range of industrial applications in detergents, shampoo, cosmetics, and surfactant formulations. The amidation reaction of Mucor miehei lipase Lipozyme IM20 was investigated for direct amidation of triacylglycerol in organic solvents. The process parameters (reaction temperature, substrate molar ratio, enzyme amount) were optimized to achieve the highest yield of anilide. The maximum yield of palmitanilide (88.9%) was achieved after 24 h of reaction at 40 °C at an enzyme concentration of 1.4% (70 mg). Kinetics of lipase-catalyzed amidation of aniline with tripalmitin has been investigated. The reaction rate could be described in terms of the Michaelis-Menten equation with a Ping-Pong Bi-Bi mechanism and competitive inhibition by both the substrates. The kinetic constants were estimated by using non-linear regression method using enzyme kinetic modules. The enzyme operational stability study showed that Lipozyme IM20 retained 38.1% of the initial activity for the synthesis of palmitanilide (even after repeated use for 48 h). Palmitanilide, a fatty acid amide, exhibited potent antimicrobial activity toward Bacillus cereus.

Antimicrobial profile of Salmonella enterica serotype Choleraesuis from free-range swine in Kakamega fish market, western Kenya.

INTRODUCTION: Salmonella enterica subspecies enterica serovar Choleraesuis is a host-adapted, facultative, intracellular pathogen that causes swine paratyphoid. Its antimicrobial resistance presents a challenge to feed manufacturing industries. However, stopping antibiotics in animal feed would have economic implications for the industry. METHODOLOGY: Conventional microbial methods for isolation and identification of S. Choleraesuis were employed. The isolates were subjected to screening against 17 antimicrobial agents and genotyping of resistance markers by PCR. The data were then analyzed and presented in percentages. RESULTS: Phenotypically, 43 out of 95 isolates showed multidrug resistance. Among the 17 antibiotics tested, resistance was observed as follows: sulphonamides (45.2%), nalidixic acid (44.25%), tetracycline (42%), ampicillin (36.8%), erythromycin (34.7%), carbenicillin (31.5%), chrolamphenical (28.4%), gentamicin (27.3%), kanamycin (24.2%), spectinomycin (21%), sulfamethoxazole-trimethoprim (16.8%), streptomycin (12.6%), cephalothion (8.4%), ofloxacin (5.2%), ciprofloxacin (4.2%), and norfloxacin (4.2%). Fifty-two isolates were susceptible to the antimicrobial agents tested. A total of 3.1% of the isolates had the integron gene pattern combination of dfrA2-aadA2 (2100 bp), dfrA12 (2100 bp); 4.2% had dfrA12-aadA2-sulI (2100 bp); 2.1% had dfrA12-aadA2 (2100 bp); and 1% had dfrA2-aadA2-sulI (2100 bp), oxa1-aadA2 (1500 bp), dfrA12-aadA2-sulI, and blaPSE (2100 bp). CONCLUSIONS: The isolated S. Choleraesuis were resistant to more than 10% of the antimicrobial agents used in this study. Appropriate surveillance is warranted to gain more information about the epidemiology, as stopping antibiotics in animal feed would have economic implications for the industry.

Diversity of pulsed-field gel electrophoresis pulsotypes, serovars, and antibiotic resistance among Salmonella isolates from wild amphibians and reptiles in the California Central Coast.

A survey of cold-blooded vertebrates and associated surface waters in a produce-growing region on the Central California Coast was done between May and September 2011 to determine the diversity of Salmonella. Samples from 460 amphibians and reptiles and 119 water samples were collected and cultured for Salmonella. Animals sampled were frogs (n=331), lizards (n=59), newts (n=5), salamanders (n=6), snakes (n=39), and toads (n=20). Salmonella was isolated from 37 individual animals, including frogs, lizards, snakes, and toads. Snakes were the most likely to contain Salmonella, with 59% testing positive followed by 15.3% of lizards, 5% of toads, and 1.2% of frogs. Fifteen water samples (12.6%) were positive. Twenty-two different serovars were identified, and the majority of isolates were S. enterica subsp. IIIb, with subsp. I, II, and IIIa also found. The serovar isolated most frequently was S. enterica subsp. IIIb 16:z10:e,n,x,z15, from snakes and frogs in five different locations. S. enterica subsp. I serovar Typhimurium and the monophasic I 6,8:d:- were isolated from water, and subspecies I Duisburg and its variants were found in animals and water. Some samples contained more than one type of Salmonella. Analysis of pulsed-field gel electrophoresis pulsotypes indicated that some strains persisted in animals and water collected from the same location. Sixty-six isolates displayed antibiotic resistance, with 27 isolates resistant to more than one antibiotic, including a subspecies IIIb isolate from snake having resistance to five different antibiotics. Twenty-three isolates were resistant to more than one class of antibiotic, and six isolates were resistant to three classes. While these subspecies of IIIa and IIIb cause fewer instances of human illness, they may serve as reservoirs of antibiotic resistance, determinants in the environment, and be sources of contamination of leafy greens associated with product recalls.

New sesquiterpenes from Pluchea arabica.

Two new sesquiterpenes, godotol A (1) and godotol B (2), were isolated from Pluchea arabica. Their structures were determined by analysis of NMR data. The absolute configurations of 1 and 2 were established by Mosher ester methodology. The godotols displayed weak activity against bacteria and the brine shrimp larvae. They were also inactive in the DPPH antioxidant assay.

Ciprofloxacin-resistant Salmonella enterica Typhimurium and Choleraesuis from pigs to humans, Taiwan.

We evaluated the disk susceptibility data of 671 nontyphoid Salmonella isolates collected from different parts of Taiwan from March 2001 to August 2001 and 1,261 nontyphoid Salmonella isolates from the National Taiwan University Hospital from 1996 to 2001. Overall, ciprofloxacin resistance was found in 2.7% (18/671) of all nontyphoid Salmonella isolates, in 1.4% (5/347) of Salmonella enterica serotype Typhimurium and in 7.5% (8/107) in S. enterica serotype Choleraesuis nationwide. MICs of six newer fluoroquinolones were determined for the following isolates: 37 isolates of ciprofloxacin-resistant (human) S. Typhimurium (N = 26) and Choleraesuis (N = 11), 10 isolates of ciprofloxacin-susceptible (MIC <1 mg/mL) (human) isolates of these two serotypes, and 15 swine isolates from S. Choleraesuis (N = 13) and Typhmurium (N = 2) with reduced susceptibility to ciprofloxacin (MIC >0.12 microg/mL). Sequence analysis of the gryA, gyrB, parC, parE, and acrR genes, ciprofloxacin accumulation, and genotypes generated by pulsed-field gel electrophoresis with three restriction enzymes (SpeI, XbaI, and BlnI) were performed. All 26 S. Typhimurium isolates from humans and pigs belonged to genotype I. For S. Choleraesuis isolates, 91% (10/11) of human isolates and 54% (7/13) of swine isolates belonged to genotype B. These two genotypes isolates from humans all exhibited a high-level of resistance to ciprofloxacin (MIC 16-64 mg/mL). They had two-base substitutions in the gyrA gene at codons 83 (Ser83Phe) and 87 (Asp87Gly or Asp87Asn) and in the parC gene at codon 80 (Ser80Arg, Ser80Ile, or Ser84Lys). Our investigation documented that not only did these two S. enterica isolates have a high prevalence of ciprofloxacin resistance nationwide but also that some closely related ciprofloxacin-resistant strains are disseminated from pigs to humans.

Direct identification and susceptibility testing of enteric bacilli from positive blood cultures using VITEK (GNI+/GNS-GA).

OBJECTIVE: To study the possibility of reporting results of identification and susceptibility testing of Gram-negative bacilli the same day as bacteremia is detected by using direct inoculation from positive blood cultures (Bactec 9240) into VITEK GNI+ and GNS-GA cards. METHODS: All blood cultures with Gram-negative enteric bacillus-like morphology on microscopy found to be positive on workdays between 15 June 1999 and 29 February 2000 were included. Identification and susceptibility testing were done by three methods: the direct method using a suspension made by differential centrifugation of positive blood culture broth for inoculation of the VITEK cards; the standard method using an inoculum made from an overnight culture on a solid media; and the routine method (reference method) using conventional testing. RESULTS: Of 169 isolates, the direct method resulted in 75% correct identifications, 9% misidentifications and 17% non-identifications. All misidentified isolates were Escherichia coli, of which 80% were reported as Salmonella arizonae. Five biochemical tests yielded most of the aberrant results; correcting the citrate and malonate reactions in most cases led to correct identification by the VITEK database. Despite a negative H2S reaction, 11 E. coli isolates were reported as S. arizonae. Two-thirds (69%) of identifications were reported within 6 h, and 95% of these were correct. The direct susceptibility testing method was assessable for 140 isolates. Correct results were found in 99% of isolate-antimicrobial combinations, and 85% were reported within 6 h. CONCLUSION: The direct VITEK method could correctly report identifications and susceptibility patterns within 6 h, making same-day reporting possible for almost two-thirds (63%) of bacteremic episodes with Gram-negative bacilli. These results could probably be improved by modification of the identification algorithms of the VITEK software.

Prevalence of multi-drug resistant Salmonella typhi among clinically diagnosed typhoid fever patients in Lagos, Nigeria.

A total of 635 clinically diagnosed typhoid fever patients were bled from three different health institutions in the metropolis of Lagos, Nigeria over a period of 15 months, May 1997 to July 1998. Out of the total blood cultured, 101 (15.9%) isolates of Salmonella species were isolated of which 68 (67.3%) were S. typhi, 17 (16.8%) and 16 (15.8%) were S. paratyphi A. and S. arizonae respectively. The overall isolation rate of S. typhi among patients is 10.7%, with most isolates 45.9% found among the severely-ill young adults, age group 16-30 years. All isolates were subjected to anti-microbial susceptibility testing using 12 different antibiotics: chloramphenicol, ampicillin, cotrimoxazole, gentamicin, colistin sulfate, nalidixic acid, nitrofurantoin, cefotaxime, tetracycline, streptomycin, ofloxacin and ciprofloxacin. All the S. typhi and S. paratyphi A isolates showed resistance to two or more of the 10 of 12 antibiotics tested particularly the 3-first-line antibiotics commonly used (chloramphenicol, ampicillin and cotrimoxazole) in the treatment of typhoid fever in Nigeria. No isolate showed resistance to ofloxacin and ciprofloxacin, however, nalidixic acid and gentamicin showed a moderate and appreciable inhibition to most of our isolates.

Salmonella arizonae in the United Kingdom from 1966 to 1990.

Salmonella arizonae are rarely isolated in the UK. Since 1966 there have been sixty-six isolates from humans of whom 35% gave a recent history of foreign travel. Terrapins and snakes are potential sources of infection.

Effects of kanamycin administration to poultry on the proliferation of drug-resistant Salmonella.

Four experiments were conducted to examine the relationship between antibiotic administration to poultry and the in vivo proliferation of Salmonellae. The frequency of isolation of drug-resistant transconjugant S. arizonae from the livers of chicks inoculated per os with multiply drug-resistant Escherichia coli and drug-sensitive S. arizonae was directly related to the concentration of kanamycin administered to the chicks in their drinking water. Kanamycin administration was also associated with a significant (P less than .05) increase in the frequency of isolation of drug-resistant transconjugant S. typhimurium from the intestines and livers of poults inoculated with drug-sensitive S. typhimurium and multiply drug-resistant E. coli. Kanamycin administration significantly reduced the spread of drug-sensitive S. typhimurium to the livers of poults inoculated only with that strain. These experiments demonstrate that antibiotic administration to poultry can enhance the proliferation of drug-resistant Salmonella.

In vivo transfer of antibiotic resistance to a strain of Salmonella arizonae.

Two trials were conducted to determine the transferability of antibiotic resistance in vivo between two strains of enteric bacteria. Newly hatched turkey poults were inoculated per os with a strain of Escherichia coli resistant to kanamycin, tetracycline, and ampicillin, a strain of Salmonella arizonae resistant to nalidixic acid and streptomycin, or both strains. Kanamycin was added to the drinking water of some poults. To test for S. arizonae cells which had received transferable resistance determinants in vivo from the E. coli strain, samples were collected by swabbing the recta of the poults and by removing segments of the intestines and livers after the birds were sacrificed. Nalidixic acid was added to the isolation media to prevent in vitro transfer from occurring after the samples were collected. Salmonella arizonae resistant to nalidixic acid, streptomycin, kanamycin, tetracycline, and ampicillin were isolated from 20% of the rectal samples taken from poults that had received both bacterial strains. S. arizonae cells which had received resistance determinants in vivo were also isolated from 73% of the intestinal samples and 8% of the liver samples taken from birds inoculated with both donor E. coli and recipient S. arizonae. Salmonella arizonae demonstrating resistance to all five antibiotics were recovered from all intestinal samples taken from birds given kanamycin in their drinking water immediately after the last S. arizonae inoculation but from only 43% of such samples taken from birds given no kanamycin.

Incidence and in vitro acquisition of transferable drug resistance in Salmonella arizonae.

Fifty-eight cultures of Salmonella arizonae isolated in 1983 from poultry sources and 21 cultures isolated in 1972 to 1974 were screened for resistance to nalidixic acid, chloramphenicol, triple sulfa, ampicillin, tetracycline, streptomycin, and kanamycin by the standardized disc susceptibility method. Sensitivity to all seven drugs was observed in 51% of the cultures tested (76% of the older cultures and 41% of the recent isolates). Resistance to two or more drugs was observed in 36% of the cultures tested (14% of the older cultures and 43% of the newer ones). The highest incidence of resistance was to streptomycin. Salmonella arizonae cultures sensitive to all seven drugs were tested for the ability conjugatively to acquire kanamycin, streptomycin, tetracycline, and ampicillin resistance determinants from a multiply resistant strain of Escherichia coli K12NA. Complete or intermediate resistance to all antibiotics was transferred to 93% of the sensitive cultures at 41 C and 85% at 28 C. Kanamycin resistance was transferred most frequently, and ampicillin resistance least frequently.

Effective control of a gentamicin-resistant Salmonella arizonae infection in turkey poults.

A gentamicin-resistant Salmonella arizonae isolate was identified as the cause of an unusually high early mortality rate in several flocks of poults produced by a primary turkey breeder. The company routinely dipped its hatching eggs in 500 ppm gentamicin before incubation and injected each poult at 1 day of age with 1 mg gentamicin. Mortality was reduced to normal, but S. arizonae was not eliminated by injecting the day-old poults with higher doses of gentamicin. S. arizonae was not isolated from sample normal-sized poults in treated groups when tetracyclines were used for antibiotic inoculation of day-old poults. Tetracyclines seemed to be completely effective only when a 5-mg subcutaneous injection per day-old poult was combined with an approximately equal dose in drinking water daily for 4 days, and therapy was accompanied by the culling of runts and other debilitated poults.

R plasmid-mediated gentamicin resistance in salmonellae isolated from turkeys and their environment.

Gentamicin-resistant salmonellae were isolated from turkey poults, hatching eggs, and litter on three unrelated farms and from a scrub jay (Aphelocoma coerulescens) on a fourth farm unrelated to the other three. The isolates were Salmonella arizonae from three of the farms (poults, eggs, scrub jay) and S. thompson (litter) from the fourth farm. The genes responsible for gentamicin resistance were located on the same conjugal plasmid. This plasmid also encoded resistance to kanamycin, streptomycin, ampicillin, and sulfadiazine.

Development of high heat resistance of Arizona neotype by preincubation at 35 degrees C in media containing NaCl.

When cells of Arizona neotype were preincubated at 35 degrees C in a medium containing NaCl (1 to 10%), they were found to be much more heat resistant upon being tested at 57 degrees C in a Trypticase (BBL Microbiology Systems) soy-yeast extract broth medium containing 10% NaCl than cells that had not been preincubated. Although no growth takes place during preincubation in the presence of 10% NaCl, some metabolic activity is necessary, since the presence of 2,4-dinitrophenol during incubation reduced the amount of gain in heat resistance. Incubation in the presence of N2, instead of air, abolished the effect. Chloramphenicol and rifampin, however, had no effect. Preincubated cells were just as heat resistant when separated by centrifugation and suspended in a fresh medium as when suspended in the incubated supernatant liquid. Conversely, the incubated supernatant liquid did not confer increased heat resistance upon unincubated cells. The increase in heat resistance, therefore, is probably a result of the salt osmotically plasmolyzing the cells by removing intracellular water, thus rendering the cellular contents more stable to heat. It is not known, however, why metabolic activity is necessary.

Plasmid-specified sucrose fermentation in Salmonella arizonae.

Thirty cultures of Salmonella arizonae 47:r:253 (Ar 23:24-25) were isolated over 7 months from the faeces of a captive reptile. All were unusual in their inability to produce a positive o-nitrophenyl-beta-D-galactosidase reaction, and in their ability to ferment sucrose. These S. arizonae carried a plasmid having a molecular mass of 72 megadaltons which specified tetracycline resistance and a plasmid of 5 megadaltons which coded for the ability to ferment sucrose. The small size of this sucrose plasmid clearly distinguishes it from others which have been reported.