Animal agriculture exposures among Minnesota residents with zoonotic enteric infections, 2012-2016.

Prospective, population-based surveillance to systematically ascertain exposures to food production animals or their environments among Minnesota residents with sporadic, domestically acquired, laboratory-confirmed enteric zoonotic pathogen infections was conducted from 2012 through 2016. Twenty-three percent (n = 1708) of the 7560 enteric disease cases in the study reported an animal agriculture exposure in their incubation period, including 60% (344/571) of Cryptosporidium parvum cases, 28% (934/3391) of Campylobacter cases, 22% (85/383) of Shiga toxin-producing Escherichia coli (STEC) O157 cases, 16% (83/521) of non-O157 STEC cases, 10% (253/2575) of non-typhoidal Salmonella enterica cases and 8% (9/119) of Yersinia enterocolitica cases. Living and/or working on a farm accounted for 61% of cases with an agricultural exposure, followed by visiting a private farm (29% of cases) and visiting a public animal agriculture venue (10% of cases). Cattle were the most common animal type in agricultural exposures, reported by 72% of cases. The estimated cumulative incidence of zoonotic enteric infections for people who live and/or work on farms with food production animals in Minnesota during 2012-2016 was 147 per 10 000 population, vs. 18.5 per 10 000 for other Minnesotans. The burden of enteric zoonoses among people with animal agriculture exposures appears to be far greater than previously appreciated.

Rapid identification of Vibrio parahaemolyticus isolated from shellfish, sea water and sediments of the Khnifiss lagoon, Morocco, by MALDI-TOF mass spectrometry.

We establish the presence of Vibrio parahaemolyticus and deepen the comparison of isolates using MALDI-TOF MS for the typing of isolates originating from the Khnifiss lagoon (Morocco). Amongst 48 samples from sea water, sediment and shellfish isolated from different sites of Khnifiss lagoon, Morocco, we obtained 22 isolates of V. parahaemolyticus identified by Vitek 2™ System (bioMérieux) and MALDI Biotyper™ (Bruker Daltonics). All isolates were highly resistant to ampicillin and ticarcillin, moderately resistant to cefalotin, but sensitive to 16 other antimicrobials tested. MALDI-TOF MS was used to discriminate between closely related environmental strains of V. parahaemolyticus. A clustering and distribution based on MALDI-TOF spectra were generated using the BioTyper 1.1™ software. Despite low diversity in regard to the biochemical characteristics and antimicrobial resistance, the isolates evoke a larger biodiversity when analysed through mass spectra of abundant proteins. Different evaluations of a cut-off value showed that, when placed at a 10% threshold of the whole diversity, isolates differed by at least three mass peaks.

[Comparative study of Vitek-2™ AIX versus Vitek-2™ PC: phenotypic resistance of enterobacteriaceae to ß-lactams]. / Comparaison du Vitek-2™ AIX versus Vitek-2™ PC par l'étude des phénotypes de résistance des entérobactéries aux ß-lactamines.

AIM OF THE STUDY: Vitek-2™ AIX versus Vitek-2™ PC have different rules for phenotypic interpretation. The aim of this study is to ensure that the raw results determined by these two versions of Vitek-2™ allow biologists to conclude to the same resistance phenotype, but also to evaluate their own phenotypic interpretation system (advanced expert system). MATERIAL AND METHODS: A total of 251 strains of Enterobacteriaceae of different groups and phenotypes was tested. Each strain was studied simultaneously on both types of Vitek-2™ from the same calibrated inoculum. We then compared their resistance phenotype to beta-lactams. RESULTS: For strains not producing ESBL or CHN, the biologist concluded in 99.3% of cases to the same resistance phenotype by interpreting the raw results of Vitek-2™ AIX versus PC. The phenotypic interpretation of biologist is different from the Vitek-2™ in respectively 40% versus 43% of cases for AIX and PC versions. For multi-resistant strains, the biologist concluded in 100% of cases to the same resistance phenotype by interpreting the raw results of Vitek-2™ AIX versus PC. In 51.5% of cases the biologist use the disk diffusion method (DD). The results of this technique put forward 29% discrepancy with the two types of Vitek-2™. Finally, when Vitek-2™ claims the presence of an ESBL alone, this result is routinely confirmed by DD. CONCLUSION: The switch from Vitek-2™ AIX to Vitek-2™ PC does not alter the results of the phenotypic interpretation of biologist.

Antibody prevalence of low-pathogenicity avian influenza and evaluation of management practices in Minnesota backyard poultry flocks.

Low-pathogenicity avian influenza (LPAI) viruses have caused illness in poultry and humans with poultry contact. To determine whether there is evidence of exposure to avian influenza viruses (AIV) among backyard poultry in Minnesota and their human caretakers, 150 flocks of backyard birds were sampled for antibodies to AIV from August 2007 through December 2008. One hundred flocks were tested through routine slaughter surveillance by the Minnesota Board of Animal Health and an additional 50 flocks were contacted and sampled by study investigators. Blood was collected from 10 to 13 birds from each flock and a survey of biosecurity and management practices was administered to the flock owner. Blood samples were tested by agar gel immunodiffusion (AGID) for influenza A antibodies. Tested flocks had a median flock size of 100 birds (range: 12-800 birds), and were most commonly owned for meat for personal use (81% of respondents), fun or hobby (58%) and eggs for personal use (56%). Although 7% of flock owners reported that their birds had shown respiratory signs in the previous 3 months, only 1 of 150 flocks tested positive for influenza by AGID. Antibodies to LPAI H6N1 were detected in the positive flock. The owner of the positive flock did not have antibodies to H6 or other common AIV. Based on the findings of this study, the risk of transmission of LPAI viruses from backyard poultry to owners in Minnesota appears to be low under current conditions and management practices.

Evaluation of a new chromogenic medium, chromID Vibrio, for the isolation and presumptive identification of Vibrio cholerae and Vibrio parahaemolyticus from human clinical specimens.

The aim of this study was to evaluate the performance of the chromID Vibrio medium for the detection of Vibrio cholerae and V. parahaemolyticus in stool and swab specimens in comparison with thiosulfate citrate bile salts sucrose (TCBS) medium. A total of 96 samples including 30 fresh stool, 32 stool, and 34 swab specimens originating from routine laboratories were tested. All samples were seeded on both media, the TCBS medium and the chromID Vibrio, directly and after an enrichment step on alkaline peptone water. Of the 96 samples studied, 34 were positive for V. cholerae and 30 were positive for V. parahaemolyticus. The sensitivity for the isolation of V. cholerae in fresh stool specimens was identical for both media, 78.5% and 100% before and after enrichment, respectively. However, positive test with chromID Vibrio concluded immediately to the presence of V. cholerae. In the case of artificial contaminations, the sensitivity of chromID Vibrio was more important than TCBS after enrichment for V. cholerae and for V. parahaemolyticus before and after enrichment. In fresh stool specimens, the specificity of chromID Vibrio for screening V. cholerae was significantly higher than TCBS (100% and 100% compared to 50% and 50% before and after enrichment, respectively) and was important for V. parahaemolyticus (100% chromID Vibrio; 93.33% TCBS).

Tularaemia in Minnesota: case report and brief epidemiology.

The 2008 case presented here of tularaemia in a cat and its owner occurred in an urban setting and was associated with animal contact, a relatively rare mode of transmission in Minnesota in recent years. Response to this case exemplified a 'One Health' approach involving pre-existing relationships, cooperation between multiple disciplines and laboratory infrastructure that facilitated information sharing.

Virulence factors produced by strains of Staphylococcus aureus isolated from urinary tract infections.

Staphylococcus aureus infections are widely prevalent in West Africa and are often associated with urinary tract infections (UTIs). Virulence factors from S. aureus have rarely been described for such infections. The purpose of the current study was to determine the prevalence of toxins and adhesion factors obtained from S. aureus isolated from presumed primary UTIs at the Cotonou University Hospital (CUH) in Benin as compared with the Strasbourg University Hospital (SUH) in France. Both ambulatory and hospitalised patients were included in the study. Sixty-five independent strains of S. aureus from CUH and 35 strains from SUH were obtained over a four-month period. Virulence factors were characterised by immunodetection or multiplex polymerase chain reaction, and meticillin susceptibility was recorded. Approximately 50% of all isolates produced at least one enterotoxin. No isolate from SUH produced Panton-Valentine leucocidin (PVL), whereas 21.5% of the S. aureus isolates from CUH produced PVL (P<0.01). Six of 14 (43%) PVL-positive isolates were meticillin-resistant. At SUH, the incidence of MRSA (57%) was significantly higher (P<0.01) than at CUH (14%). Genes encoding clumping factor B, and elastin and laminin binding proteins were detected in almost all isolates (80%), irrespective of the geographical origin. The results for elastin binding protein differed significantly from published data regarding isolates from other clinical origins. Staphylococcal toxins and adhesion factors may be important in the physiopathology of UTI.

Recurrent outbreaks of cryptosporidiosis associated with calves among students at an educational farm programme, Minnesota, 2003.

Enteric illness outbreaks among middle-/high-school students in consecutive semesters of an educational farm programme were investigated with retrospective cohort studies. During the first outbreak, 31/92 (34%) interviewed students were ill. Risk factors included participating in animal science class (RR 8.1, 95% CI 1.2-55.2) and contact with calves (RR 4.2, 95% CI 1.1-16.2). Stool samples from seven students and two calves yielded Cryptosporidium parvum. Students cared for animals in street clothes and practised poor hand washing. During the second outbreak, 37/81 (46%) interviewed animal science students were ill. Risk factors included having visible manure on hands, and wearing coveralls and boots. Stool samples from seven students and eight calves yielded C. parvum. Student hand washing was still inadequate. Coveralls/boots were cleaned infrequently and removed after hand washing. These outbreaks of cryptosporidiosis resulted from calf contact and inadequate hygiene practices. The failure to adequately implement recommended interventions contributed to the second outbreak.

External quality assessment for molecular detection of Bordetella pertussis in European laboratories.

Although the PCR for the detection of Bordetella pertussis is routinely performed in diagnostic laboratories, no quality assessment program has so far been described. We report on the results obtained with two external quality assessment proficiency panels sent to European laboratories. The first proficiency panel contained a series of dilutions of three previously characterized B. pertussis clinical isolates and two negative controls. No false-positive results were reported by six laboratories providing seven data sets. The reported limits of detection of the three B. pertussis strains varied between 4 and 4,000, 9 and 9,000, and 3 and 30,000 CFU/ml, respectively. The second proficiency panel, composed of a series of dilutions of reference strains of B. pertussis, B. holmesii, B. hinzii, and B. bronchiseptica, as well as negative controls, was sent to nine laboratories. One laboratory reported a negative result for a sample and reported a B. parapertussis-positive sample to be positive for B. pertussis. By using the B. pertussis-specific target gene pertactin, one laboratory detected B. pertussis with 100% specificity. All other laboratories, which used IS481-based assays, reported positive results for the samples containing B. holmesii and B. bronchiseptica, species that have occasionally been recovered from human respiratory samples. These data show that the choice of the target gene is particularly critical for the species specificity of B. pertussis PCR assays.

Predominant Staphylococcus aureus isolated from antibiotic-associated diarrhea is clinically relevant and produces enterotoxin A and the bicomponent toxin LukE-lukD.

Staphylococcus aureus was isolated as the predominant or only isolate from cultures of stools of 60 patients over 2 years in a university hospital, leading to the collection of 114 isolates. Diarrhea was observed in 90% of the patients. Ninety-eight percent of the patients had received antibiotics in the month before the diarrhea. Ninety-two percent of the S. aureus isolates were methicillin resistant. S. aureus was encountered with antibiotic-associated diarrhea among 47 quite elderly patients affected or not affected by a gastrointestinal disease. Among the antimicrobial treatments, cessation of the previous therapy when possible or rapid application of oral vancomycin therapy was the most appropriate. Analysis of total DNA by pulsed-field gel electrophoresis revealed 27 different SmaI pulsotypes distributed in 15 clusters. The pulsotypes never differed for related isolates from a single patient, even if they originated from patients with bacteremia. S. aureus was not isolated as the predominant isolate in cultures of stools of 57 patients who received an antimicrobial treatment for more than 5 days without diarrhea. Occurence of production of both enterotoxin A and the bicomponent leucotoxin LukE-LukD by the S. aureus isolates was significantly different from that by random isolates. The results strongly suggest that when predominant in stool samples, S. aureus should be considered a possible etiologic agent for some cases of antibiotic-associated diarrhea.

[Evaluation of Hemofast MRSA for identification of Staphylococcus aureus and detection of methicillin-resistance staphylococci, directly in blood culture broths]. / Evaluation d'Hémofast MRSA pour l'identification de Staphylococcus aureus et la détection des staphylocoques résistants à la méticilline, directement à partir des bouillons d'hémocultures.

The newly developed Hemofast MRSA system for Staphylococcus aureus identification and methicillin-resistant staphylococci (MRS) detection in blood culture broths was evaluated in 106 Bactec broths containing grapelike clusters of Gram-positive cocci. All 26 S. aureus positive broths were correctly identified by Hemofast MRSA within two hours, and 81% were identified within one hour. Sensitivity and specificity were both 100%. Accuracy of MRS detection was 100%, i.e., no discrepancies versus the agar diffusion method were found. When the 28 broths containing coagulase negative staphylococci (CNS) were tested using an inoculum ten fold larger than for S. aureus testing, a number of discrepancy were recorded. For 49 other broths containing CNS, accuracy was 98.5% when the test was interpreted based on the growth rate of the strain, according to the manufacturer's instructions. One broth containing S. epidermidis strain susceptible yielded a false result with Hemofast MRSA, indicating that it probably contained a contaminant. The other discrepancies occurred with specimens containing mixed populations with CNS or a Micrococcus strain. Most (85%) results were available within 4 hrs 30 min, irrespective of the S. aureus or CNS strains. Avoiding the isolation step on agar, Hemofast MRSA saves 24 to 48 hours, thus allowing earlier antistaphylococcal treatment.

[Identification of Staphylococcus aureus within 2 hours in blood culture broths]. / Identification en 2 heures de Staphylococcus aureus dans les bouillons d'hémocultures.

Staphylococcus aureus identification is one of the priorities of the microbiological diagnosis of the staphylococcal infections. Current identification methods are carried out after a first step of colony isolation on agar media. We describe a fluorogenic method for S. aureus identification, which is directly applied to blood culture broths. This method uses a gel tube which allows an optimized microculture of the bacteria. 129 clinical samples of blood cultures (HEC) containing gram positive cocci in grapelike clusters (35 Vital bottles, 94 Bactec bottles), and 77 inoculated blood culture (HE) with collection strains of S. aureus are included in the study. Bacteria are concentrated and separated from other components of sample in the gel tube. Staphylococci are revealed during a microculture in the gel phase, by using a colorimetric substrate of their dehydrogenases. Then, staphylococci are recovered in an adapted culture medium containing human prothrombin and a fluorogenic substrate, which is specific for the staphylocoagulase. After 1 to 2 h incubation at 37 degrees C, a blue fluorescence shows the presence of S. aureus. Among the 40 HEC containing S. aureus the test is positive for 37 samples. For 3 cases, the test is not interpretable, due to non lysis red blood cells in the gel phase of the tube. No false positive result is observed for the HEC containing coagulase-negative staphylococci. Moreover, our method is positive with the 77 HE. 94.7% of tested samples (HEC and HE) show a fluorescence after only one hour and half. Sensibility and specificity are both 100%. We propose a rapid method for S. aureus identification directly applied to blood culture broths. This method saves 24 h, avoiding the isolation step on agar. Therefore, the treatment of staphylococcal infections and possible isolation measures could earlier set up.

Analysis with a combination of macrorestriction endonucleases reveals a high degree of polymorphism among Bordetella pertussis isolates in eastern France.

From 1990 to 1996, routine screening for whooping cough identified 399 patients with a calmodulin-dependent adenylate cyclase-positive test result and yielded 69 Bordetella pertussis isolates. None of the patients were fully vaccinated, and most were less than 6 months old. Analysis of total DNA by pulsed-field gel electrophoresis (PFGE) after XbaI, SpeI, or DraI macrorestriction yielded 19, 15, and 5 different patterns, respectively, whereas ribotyping failed to demonstrate any strain polymorphism. Discrimination among the isolates was improved by combining the PFGE profiles. Some patterns were more frequent, but the corresponding patients were not clearly epidemiologically related. The patterns for two strains obtained during a 3-month period from patients who were neighbors differed by the length of a single DNA fragment. These data strongly suggest that one type of isolate is widely spread throughout the world and is carried by individuals other than patients who develop a true illness.

A new gel tube method for the direct detection, identification and susceptibility testing of bacteria in clinical samples.

We recently developed a simple new method which is designed to separate and concentrate bacteria from a sample by centrifugation in a gel system. Bacterial enzyme activity is then detected inside the gel without further manipulation using a colorimetric or fluorogenic substrate. The method provides a rapid, direct means of detecting bacteria in clinical samples, dispensing with the 24-h period normally required to isolate colonies on agar. Various applications of the method are described below, e.g. screening of negative urine samples, identification of Escherichia coli in urine samples, identification of Staphylococcus aureus in blood culture broths and detection of oxacillin-resistant S. aureus in blood culture broths. The advantages of the gel system and other applications are discussed.

[Tricentric study of the sensitivity of Pseudomonas aeruginosa serotyping to beta-lactams and aminoglycosides]. / Etude tricentrique de la sensibilité des sérotypes de Pseudomonas aeruginosa aux beta-lactamines et aux aminosides.

The aim of this study was to evaluate the susceptibility of 300 Pseudomonas aeruginosa strains to 6 beta-lactams and to 2 aminoglycosides and to compare this susceptibility according to the serotype, the sample nature, the unit activity and the geographic situation of the hospital. The susceptibility level was ascertained using the disk method. Serotyping of the strains was also performed. Susceptibility level was 67% for ticarcillin, 82% for piperacillin-tazobactam combination, 85% for ceftazidim, 81% for cefepim, 74% for aztreonam, 83% for imipenem, 73% for tobramycin and 86% for amikacin. Non-serotypable strains represented 20.6%. The most common serotypes were O6 (19%) and O11 (13%). Discrepancies between cefepime and ceftazidime were found in 15 strains after MIC determination, 6 of them were O12 strains. It appears that strain susceptibility was function of the serotype distribution. Cefepime exhibited outstanding activity against P. aeruginosa, similar to ceftazidim activity.

[Role of hygiene and bacteriological laboratories in the management of an epidemic of Enterobacter aerogenes multiresistant to antibiotics]. / Rôles des laboratoires d'hygiène et de bactériologie dans la prise en charge d'une épidémie à Enterobacter aerogenes multirésistant aux antibiotiques.

We describe a multiresistant Enterobacter aerogenes outbreak in an intensive care-unit. An epidemiology study based on phenotypic characters (species diagnosis and antibiotype) was completed by a genotypic study (pulsed field electrophoresis) to confirm bacterial clonality. The hygiene laboratory proposed numerous preventive measures to limit bacterial dispersion. We describe the role of bacteriologists, hygienists and medical staff to stop the bacterial dispersion.

[In vitro antibacterial activity of a new fluoroquinolone, sparfloxacin, against nosocomial bacteria, and concordance curve]. / Activité anti-bactérienne in vitro d'une nouvelle fluoroquinolone, la sparfloxacine sur les bactéries hospitalières et courbe de concordance.

Minimal inhibitory concentrations (MICs) of sparfloxacin (SFX) were determined by agar dilution for 3,164 bacterial strains isolated in 10 university hospitals; in addition, antibiograms by agar diffusion were performed with 5 micrograms disks. Activity of SFX against nalidixic acid (NAL) susceptible (S) Enterobacteriaceae was close to that of other fluoroquinolones (FQ) (MIC 50 and 90: 0.06-0.5 microgram/ml); like for other FQ, this activity was reduced against NAL intermediate and resistant (R) Enterobacteriaceae (2-16). MICs of SFX against P. aeruginosa were between 0.12 and 16 (1-32). SFX had also a good activity against NAL-S A. baumannii (CMI < or = 0.25) but this activity is reduced against NAL-R Acinetobacter (16). SFX was highly active against Haemophilus (0.016-0.06) gonococci (0.008), meningococci (0.008) and B. catarrhalis (0.008-0.03). SFX showed activity superior to the currently available FQ against methicillin susceptible staphylococci (0.06); the resistant strains [8] are usually methicillin resistant. SFX is more effective against enterococci (0.5), streptococci (0.25-0.5) and particularly pneumococci (0.25-0.5) including penicillin-resistant strains. The coefficient correlation of the regression curve is 0.876; for MIC breakpoints of 1 and 2 micrograms/ml, zone diameter breakpoints should be 20 and 16 mm.

[Mobile species of the genus Aeromonas: difficulties of identification and pathogenicity]. / Les espès mobiles du genre Aeromonas: difficultés de l'identification et pouvoir pathogène.

Sixty-two Aeromonas strains (39 of clinical and 23 of environmental origin) were identified. The suicide phenomenon and autoagglutination were studied. Identification is based on esculin hydrolysis; fermentation of arabinose salicin, sucrose and mannitol; gas production from glucose, indole and beta hemolysis; Voges-Proskauer and decarboxylation reactions; and finally resistance to cephalothin (30 micrograms) and colistin (4 micrograms/ml). Thirty-four per cent of A hydrophila, 33% A caviae, 28% A veronii subspecies sobria, 3% A jandaei, 2% A veronii subspecies veronii were accurately identified. Also, several new species were identified such as A trota, A enteropelogenes, A schubertii, A ichthiosmia, according to the more recently proposed taxa. This identification scheme could enhance our knowledge concerning virulence factors, pathogenicity and environmental distribution.

Decreased lipolytic activity in tissues during infectious and inflammatory stress.

The clearance rate of endogenous and exogenous circulating lipids during the septic or inflammatory state remains a controversial subject. Thus, we have developed rat models of gram-negative and gram-positive sepsis and of sterile inflammation to study this problem. In addition to the febrile response, these stresses induced some of the following metabolic changes in the blood: decreased total protein, albumin, and ketone body levels and increased lactate, pyruvate, alanine, cholesterol, and triacylglycerol levels. The activities of heart, diaphragm, and adipose tissue lipoprotein lipase and of hepatic lipase decreased to differing extents depending on whether the enzyme substrate was a long-chain or a medium- and long-chain triglyceride-based emulsion. However, the latter emulsion was always hydrolyzed faster than the former. This observation suggests that, during infection/inflammation, the medium- and long-chain triglyceride-based emulsion would be cleared more quickly, would induce less hypertriglyceridemia, and would thus deliver lipid energy more rapidly than a traditional long-chain triglyceride-based emulsion.

Gram-negative bacteria sepsis in the rat and tissue lipolytic activity on LCT and MCT/LCT-based commercial parenteral emulsions.

The aim of this study was to evaluate the effect of a gram-negative bacteria sepsis on the activity of the enzymes lipoprotein lipase (LPL) and hepatic lipase (HL), involved in the clearance of circulating triacylglycerol-rich fat particles. Fasting rats were intravenously injected with NaCl9 g.l-1, live or heat-killed Pseudomonas aeruginosa bacteria. After 18 h the animals were killed. When compared to controls, the 2 treated groups showed an increase in body temperature, cholesterolemia, triglyceridemia and a decrease in ketonemia, proteinemia, albuminemia and in the in vitro activity of diaphragm, heart and adipose tissue LPL and of HL. The decrease in the enzyme activities occurred independent of the type of emulsion used as in vitro substrate, whether it was based on long-chain triglycerides or on medium- and long-chain triglycerides, but in any case the activity was lower with the first than with the second type of fat emulsion.