M. Jahangir Alam

Prevalence and antimicrobial resistance of Salmonella in retail foods in northern China

A total of 387 retail meat, seafood and milk powder samples were collected from nine cities in northern China in 2005 and screened for the presence of Salmonella. Salmonella strains isolated were subjected to serotyping and antimicrobial susceptibility testing. Salmonella was isolated from 81 (20.9%, 81/387) samples and classified into 23 serotypes. The isolates were frequently resistant to sulfamethoxazole (86.4%), sulfamethoxazole/trimethoprim (48.1%), nalidixic acid (30.9%), tetracycline (19.8%), carboxybenzylpenicillin (17.3%), amoxicillin (17.3%) and ampicillin (16.0%). The multiple resistance (resistance to ≥ 3 antibiotics) was found in 29.6% (n = 24) isolates. Additionally, 4 isolates from chicken displayed the ACSSuTNx profile, resistant to ampicillin, chloramphenicol, streptomycin, sulfonamide, tetracycline and nalidixic acid, in particular, strain HBS084 showing the resistance to as many as 20 antibiotics. Salmonella from chicken showed the higher frequency of antimicrobial resistance. Our findings indicate that in northern China food products of animal origin can be a source of exposure for consumers to multiresistant Salmonella strains.

Comparison of the T2Dx instrument with T2Candida assay and automated blood culture in the detection of Candida species using seeded blood samples.

As a delay in the diagnosis and treatment of candidemia is associated with increased mortality and healthcare costs, a more rapid method of detection is urgently needed. The T2Candida assay is a new rapid diagnostic test, which uses T2 magnetic resonance technology to identify Candida spp. directly from whole blood in approximately 3 hours. In this study, the performance of the BACTEC 9050 using Aerobic Plus/F blood culture bottles was compared to that of the T2Candida assay run on the T2Dx Instrument for detection of Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, and Candida krusei, in seeded blood samples at concentrations between 3.1 and 11 CFU/mL. The BACTEC 9050 detected Candida growth in 100% of bottles (n = 20 replicates) within 5 days for all species (63.23 ± 30.27 hours), with the exception of Candida glabrata (0%). The T2Candida assay had a 100% detection rate for each species (n = 13-20 replicates) within 3 hours including C. glabrata. The sensitivity and specificity of the T2Candida assay were 1 and 0.978, respectively.

Treatment of Candida famata bloodstream infections: case series and review of the literature

OBJECTIVES Candida famata (also known as Debaryomyces hansenii and Torulopsis candida) is a commensal yeast found in cheese, dairy products and the environment. C. famata accounts for 0.2%-2% of invasive candidiasis. The purpose of this study was to provide an overview of the treatment of C. famata bloodstream infections. METHODS The clinical course of two hospitalized patients who developed C. famata fungaemia within 2 weeks of each other was summarized along with available data regarding in vitro susceptibility patterns, genotyping and clinical outcomes of these cases compared with the published literature. RESULTS AND CONCLUSIONS C. famata appears to exhibit reduced susceptibility to echinocandins and azoles, particularly in the setting of prior antifungal exposure. The removal of indwelling central venous catheters and prompt initiation of therapy with liposomal amphotericin B is recommended for successful treatment of C. famata fungaemia, particularly in immunocompromised patients. These cases also help provide justification for routine antifungal susceptibility testing in patients with candidaemia to guide optimal antifungal therapy.

Rapid detection of food-borne Listeria monocytogenes by real-time quantitative loop-mediated isothermal amplification

The purpose of this study was to develop a real-time quantitative loop-mediated isothermal amplication (LAMP) method for the rapid, sensitive, and convenient detection of Listeria monocytogenes in food. The LAMP method could amplify the hlyA gene of L. monocytogenes successfully at 63°C with a loopamp real-time turbidimeter. The detection limits of the LAMP for hlyA gene were 6 colony forming units (CFU)/tube. A standard curve was generated for L. monocytogenes LAMP by plotting the graph based different log CFU values of L. monocytogenes and time of positivity through real-time monitoring of the amplication. Then, the LAMP method was employed to test 94 retail food samples effectively. Sensitivity in detection of L. monocytogenes by the LAMP was higher than that of PCR and none of the conventional methodpositive samples was missed by the LAMP method.

Impact on toxin production and cell morphology in Clostridium difficile by ridinilazole (SMT19969), a novel treatment for C. difficile infection.

Objectives Ridinilazole (SMT19969) is a narrow-spectrum, non-absorbable antimicrobial with activity against Clostridium difficile undergoing clinical trials. The purpose of this study was to assess the pharmacological activity of ridinilazole and assess the effects on cell morphology. Methods Antibiotic killing curves were performed using the epidemic C. difficile ribotype 027 strain, R20291, using supra-MIC (4× and 40×) and sub-MIC (0.125×, 0.25× and 0.5×) concentrations of ridinilazole. Following exposure, C. difficile cells were collected for cfu counts, toxin A and B production, and morphological changes using scanning electron and fluorescence microscopy. Human intestinal cells (Caco-2) were co-incubated with ridinilazole-treated C. difficile growth medium to determine the effects on host inflammatory response (IL-8). Results Treatment at supra-MIC concentrations (4× and 40× MIC) of ridinilazole resulted in a significant reduction in vegetative cells over 72 h (4 log difference, P < 0.01) compared with controls without inducing spore formation. These results correlated with a 75% decrease in toxin A production (P < 0.05) and a 96% decrease in toxin B production (P < 0.05). At sub-MIC levels (0.5× MIC), toxin A production was reduced by 91% (P < 0.01) and toxin B production was reduced by 100% (P < 0.001), which resulted in a 74% reduction in IL-8 release compared with controls (P < 0.05). Sub-MIC (0.5×)-treated cells formed filamentous structures ∼10-fold longer than control cells. Following fluorescence labelling, the cell septum was not forming in sub-MIC-treated cells, yet the DNA was dividing. Conclusions Ridinilazole had robust killing effects on C. difficile that significantly reduced toxin production and attenuated the inflammatory response. Ridinilazole also elicited significant cell division effects suggesting a potential mechanism of action.

Evaluation of Portability and Cost of a Fluorescent PCR Ribotyping Protocol for Clostridium difficile Epidemiology

ABSTRACT Clostridium difficile is the most commonly identified pathogen among health care-associated infections in the United States. There is a need for accurate and low-cost typing tools that produce comparable data across studies (i.e., portable data) to help characterize isolates during epidemiologic investigations of C. difficile outbreaks and sporadic cases of disease. The most popular C. difficile-typing technique is PCR ribotyping, and we previously developed methods using fluorescent PCR primers and amplicon sizing on a Sanger-style sequencer to generate fluorescent PCR ribotyping data. This technique has been used to characterize tens of thousands of C. difficile isolates from cases of disease. Here, we present validation of a protocol for the cost-effective generation of fluorescent PCR ribotyping data. A key component of this protocol is the ability to accurately identify PCR ribotypes against an online database (http://walklab.rcg.montana.edu) at no cost. We present results from a blinded multicenter study to address data portability across four different laboratories and three different sequencing centers. Our standardized protocol and centralized database for typing of C. difficile pathogens will increase comparability between studies so that important epidemiologic linkages between cases of disease and patterns of emergence can be rapidly identified.

In the Endemic Setting, Clostridium difficile Ribotype 027 Is Virulent But Not Hypervirulent

BACKGROUND Conflicting reports have been published on the association between Clostridium difficile ribotypes and severe disease outcomes in patients with C. difficile infection (CDI); several so-called hypervirulent ribotypes have been described. We performed a multicenter study to assess severe disease presentation and severe outcomes among CDI patients infected with different ribotypes. METHODS Stool samples that tested positive for C. difficile toxin were collected and cultured from patients who presented to any of 7 different hospitals in Houston, Texas (2011-2013). C. difficile was characterized using a fluorescent PCR ribotyping method. Medical records were reviewed to determine clinical characteristics and ribotype association with severe CDI presentation (ie, leukocytosis and/or hypoalbuminemia) and severe CDI outcomes (ie, ICU admission, ileus, toxic megacolon, colectomy, and/or in-hospital death). RESULTS Our study included 715 patients aged 61±18 years (female: 63%; median Charlson comorbidity index: 2.5±2.4; hospital-onset CDI: 45%; severe CDI: 36.7%; severe CDI outcomes: 12.3%). The most common ribotypes were 027, 014-020, FP311, 002, 078-126, and 001. Ribotype 027 was a significant independent predictor of severe disease (adjusted odds ratio [aOR], 2.24; 95% confidence interval [CI], 1.53-3.29; P<.001) and severe CDI outcomes (aOR, 1.71; 95% CI, 1.02-2.85; P=.041) compared with all other ribotypes in aggregate. However, in an analysis using all common ribotypes as individual variables, ribotype 027 was not associated with severe CDI outcomes more often than other ribotypes. CONCLUSION Ribotype 027 showed virulence equal to that of other ribotypes identified in this endemic setting. Clinical severity markers of CDI may be more predictive of severe CDI outcomes than a particular ribotype.

Prevalence and Persistence of Salmonella in Cohorts of Feedlot Cattle

Our objectives were to determine factors associated with fecal prevalence of Salmonella at feedlot entry and within 24 h of harvest (preharvest), and to assess potential persistence of Salmonella strains within cattle populations. This repeated cross-sectional study followed 5559 beef cattle within 30 feedlot cohorts. Samples (n = 30) of fresh feces were collected from the pen floor of each cohort at feedlot entry and preharvest. Samples were subjected to a selective Salmonella isolation protocol and serotypes were determined for Salmonella isolates. Genetic similarity of a subset of isolates was determined using pulsed-field gel electrophoresis (PFGE). Cattle health and performance data were recorded electronically by feedlot personnel. Cohort-level generalized linear mixed models were used to assess bivariable associations. Fecal prevalence of Salmonella within a cohort at feedlot entry (mean = 64.7%) was not associated with preharvest prevalence (mean = 72.6%). Prevalence at feedlot entry was negatively associated with mean entry weight (p = 0.02). Preharvest prevalence was positively associated with the number of days in the feedlot (p = 0.02), cumulative morbidity (p = 0.01), and cumulative mortality (p = 0.03). We recovered Salmonella isolates with identical PFGE profiles both at feedlot entry and preharvest from 14 cohorts of cattle. Fecal prevalence of Salmonella immediately before harvest may be higher in subsets of the feedlot population, but does not appear to be affected by prevalence at feedlot entry. However, PFGE subtypes of Salmonella appear to persist within and among feedlot cohorts throughout the feeding period.

Acquisition of Clostridium difficile Colonization and Infection After Transfer From a Veterans Affairs Hospital to an Affiliated Long-Term Care Facility.

BACKGROUND Clostridium difficile infection (CDI) and asymptomatic carriage of toxigenic C. difficile are common in long-term care facilities (LTCFs). However, whether C. difficile is frequently acquired in the LTCF versus during acute-care admissions remains unknown. OBJECTIVE To test the hypothesis that LTCF residents often acquire C. difficile colonization and infection in the LTCF DESIGN This 5-month cohort study was conducted to determine the incidence of acquisition of C. difficile colonization and infection in asymptomatic patients transferred from a Veterans Affairs hospital to an affiliated LTCF. METHODS Rectal swabs were cultured for toxigenic C. difficile at the time of transfer to the LTCF and weekly for up to 6 weeks. We calculated the proportion of LTCF-onset CDI cases within 1 month of transfer that occurred in residents colonized on admission versus those with new acquisition in the LTCF. RESULTS Of 110 patients transferred to the LTCF, 12 (11%) were asymptomatically colonized with toxigenic C. difficile upon admission, and 4 of these 12 patients (33%) developed CDI within 1 month, including 3 recurrent and 1 initial CDI episode. Of 82 patients with negative cultures on transfer and at least 1 follow-up culture, 22 (27%) acquired toxigenic C. difficile colonization, and 4 developed CDI within 1 month, including 1 recurrent and 3 initial CDI episodes. CONCLUSION LTCF residents frequently acquired colonization with toxigenic C. difficile after transfer from the hospital, and 3 of 4 initial CDI cases with onset within 1 month of transfer occurred in residents who acquired colonization in the LTCF. Infect Control Hosp Epidemiol 2017;38:1070-1076.

Antibiotic Resistance and Growth of the Emergent Pathogen Escherichia albertii on Raw Ground Beef Stored under Refrigeration, Abuse, and Physiological Temperature

Escherichia albertii is an emerging gram-negative facultative rod that has been implicated in multiple cases of human diarrheal disease, particularly in young children. When biochemical and other typing methods have been used, this organism has often been misidentified due to similarities with other members of the family Enterobacteriaceae. Isolates have been reported to be capable of producing attachment and effacement lesions via the synthesis of intimin, cytolethal distending toxin, and a variant form of Shiga toxin. The purposes of this study were to characterize the antibiotic resistance characteristics and the growth of individual strains of E. albertii on raw ground beef at different storage temperatures. Nalidixic acid-resistant strains of E. albertii were inoculated onto raw ground beef to a target of 4.0 log CFU/g, and samples were then aerobically incubated at 5, 22, or 35°C for various time periods prior to microbiological enumeration of the pathogen on lactose-free MacConkey agar containing 50 mg of nalidixic acid per liter and 0.5% L-rhamnose. Antibiotic resistance was determined using a broth microdilution assay. E. albertii did not grow at 5°C, with populations declining slowly over 14 days of refrigerated storage. Strains of the organism grew well under abusive storage, increasing by 2.5 to 3.1 log CFU/g and 4.1 to 4.3 log CFU/g after 24 h at 22 and 35°C, respectively. All strains were resistant to tetracycline but were sensitive to tested cephalosporins and chloramphenicol. Resistance to penicillin was observed, but susceptibility to other members of the b -lactam group, including ampicillin, amoxicillin, and clavulanic acid, was recorded. E. albertii represents an emerging pathogen with a probable foodborne transmission route. Future research should focus on verifying food process measures able to inactivate the pathogen.