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1.  Comparison of Multilocus Sequence Typing, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Typing for Characterization of Salmonella enterica Serotype Newport Isolates 
Journal of Clinical Microbiology  2006;44(7):2449-2457.
In the United States, multidrug-resistant phenotypes of Salmonella enterica serotype Newport (commonly referred to as MDR-AmpC) have emerged in animals and humans and have become a major public health problem. Although pulsed-field gel electrophoresis (PFGE) is the current “gold standard” typing method for Salmonella, multilocus sequence typing (MLST) may be more relevant to investigations exploring evolutionary and population biology relationships. In this study, 81 Salmonella enterica serotype Newport isolates from humans, food animals, and retail foods were examined for antimicrobial susceptibility and characterized using PFGE and MLST of seven genes, aroC, dnaN, hemD, hisD, purE, sucA, and thrA. Forty-nine percent of the isolates were resistant to nine or more of the tested antimicrobials. Salmonella isolates displayed resistance most often to sulfamethoxazole (57%), streptomycin (56%), tetracycline (56%), ampicillin (52%), and ceftiofur (49%) and, to a lesser extent, to kanamycin (19%), trimethoprim-sulfamethoxazole (17%), and gentamicin (11%). A total of 43 PFGE patterns were generated using XbaI, indicating a genetically diverse population. The largest PFGE cluster contained isolates from clinically ill swine, cattle, and humans. MLST resulted in 12 sequence types (STs), with one type encompassing 62% of the strains. Ten new sequence types and one novel allele type were identified. Furthermore, MLST typing showed that strains closely related by PFGE clustered in major STs, whereas more distantly related strains were separated into two clusters by PFGE. The results of this study demonstrated that the MLST scheme employed here clustered S. enterica serovar Newport isolates in distinct molecular populations, and strain discrimination was enhanced by combining PFGE, antimicrobial susceptibility, and MLST results.
PMCID: PMC1489510  PMID: 16825363
2.  Factors influencing prescribing habits. 
PMCID: PMC1371730  PMID: 1793655
4.  Study of out-of-hours visits to children 
Out-of-hours calls made to children by an urban practice were studied over a 12-month period. Out of a total of 1463 out-of-hours visits, 336 were to children, of whom 17 were admitted to hospital. The proportion of the out-of-hours calls which were to children is less than had been expected. This unexpectedly low level of out-of-hours visits to children and the type of conditions encountered should be taken into account when planning vocational training programmes.
PMCID: PMC1960259  PMID: 4057176
11.  Characterization of Salmonella enterica Serotype Newport Isolated from Humans and Food Animals 
Journal of Clinical Microbiology  2003;41(12):5366-5371.
Salmonella enterica serotype Newport isolates resistant to at least nine antimicrobials (including extended-spectrum cephalosporins), known as serotype Newport MDR-AmpC isolates, have been rapidly emerging as pathogens in both animals and humans throughout the United States. Resistance to extended-spectrum cephalosporins is associated with clinical failures, including death, in patients with systemic infections. In this study, 87 Salmonella serotype Newport strains were characterized by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing and examined for the presence of class 1 integrons and blaCMY genes. Thirty-five PFGE patterns were observed with XbaI, and three of these patterns were indistinguishable among isolates from humans and animals. Fifty-three (60%) Salmonella serotype Newport isolates were identified as serotype Newport MDR-AmpC, including 16 (53%) of 30 human isolates, 27 (93%) of 29 cattle isolates, 7 (70%) of 10 swine isolates, and 3 (30%) of 10 chicken isolates. However, 28 (32%) Salmonella serotype Newport isolates were susceptible to all 16 antimicrobials tested. The blaCMY gene was present in all serotype Newport MDR-AmpC isolates. Furthermore, the plasmid-mediated blaCMY gene was transferable via conjugation to an Escherichia coli strain. The transconjugant showed the MDR-AmpC resistance profile. Thirty-five (40%) of the isolates possessed class 1 integrons. Sequence analyses of the integrons showed that they contained aadA, which confers resistance to streptomycin, or aadA and dhfr, which confer resistance to trimethoprim-sulfamethoxazole. One integron from a swine isolate contained the sat-1 gene, which encodes resistance to streptothricin, an antimicrobial agent that has never been approved for use in the United States. In conclusion, Salmonella serotype Newport MDR-AmpC was commonly identified among Salmonella serotype Newport isolates recovered from humans and food animals. These findings support the possibility of transmission of this organism to humans through the food chain.
PMCID: PMC309039  PMID: 14662912
12.  BP Screening.. 
PMCID: PMC2383544  PMID: 21297833
13.  Antimicrobial Susceptibility Testing of Aquatic Bacteria: Quality Control Disk Diffusion Ranges for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28°C 
Journal of Clinical Microbiology  2003;41(9):4318-4323.
Quality control (QC) ranges for disk diffusion susceptibility testing of aquatic bacterial isolates were proposed as a result of a multilaboratory study conducted according to procedures established by the National Committee for Clinical Laboratory Standards (NCCLS). Ranges were proposed for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28°C for nine different antimicrobial agents (ampicillin, enrofloxacin, erythromycin, florfenicol, gentamicin, oxolinic acid, oxytetracycline, ormetoprim-sulfadimethoxine, and trimethoprim-sulfamethoxazole). All tests were conducted on standard Mueller-Hinton agar. With ≥95% of all data points fitting within the proposed QC ranges, the results from this study comply with NCCLS guidelines and have been accepted by the NCCLS Subcommittee for Veterinary Antimicrobial Susceptibility Testing. These QC guidelines will permit greater accuracy in interpreting results and, for the first time, the ability to reliably compare susceptibility test data between aquatic animal disease diagnostic laboratories.
PMCID: PMC193829  PMID: 12958263
16.  Characterization of Tn1546 in Vancomycin-Resistant Enterococcus faecium Isolated from Canine Urinary Tract Infections: Evidence of Gene Exchange between Human and Animal Enterococci 
Journal of Clinical Microbiology  2002;40(12):4659-4665.
Thirty-five enterococcal isolates were recovered from dogs diagnosed with urinary tract infections at the Michigan State University Veterinary Teaching Hospital over a 2-year period (1996 to 1998). Isolated species included Enterococcus faecium (n = 13), Enterococcus faecalis (n = 7), Enterococcus gallinarum (n = 11), and Enterococcus casseliflavus (n = 4). Antimicrobial susceptibility testing revealed several different resistance phenotypes, with the majority of the enterococcal isolates exhibiting resistance to three or more antibiotics. One E. faecium isolate, CVM1869, displayed high-level resistance to vancomycin (MIC > 32 μg/ml) and gentamicin (MIC > 2,048 μg/ml). Molecular analysis of this isolate revealed the presence of Tn1546 (vanA), responsible for high-level vancomycin resistance, and Tn5281 carrying aac6′-aph2", conferring high-level aminoglycoside resistance. Pulsed-field gel electrophoresis analysis revealed that CVM1869 was a canine E. faecium clone that had acquired Tn1546, perhaps from a human vancomycin-resistant E. faecium. Transposons Tn5281 and Tn1546 were located on two different conjugative plasmids. Sequence analysis revealed that in Tn1546, ORF1 had an 889-bp deletion and an IS1216V insertion at the 5′ end and an IS1251 insertion between vanS and vanH. To date, this particular form of Tn1546 has only been described in human clinical vancomycin-resistant enterococcus isolates unique to the United States. Additionally, this is the first report of a vancomycin-resistant E. faecium isolated from a companion animal in the United States.
PMCID: PMC154613  PMID: 12454168
17.  A Student's View 
British Medical Journal  1970;1(5697):692.
PMCID: PMC1700556
18.  Standardization of Broth Microdilution and Disk Diffusion Susceptibility Tests for Actinobacillus pleuropneumoniae and Haemophilus somnus: Quality Control Standards for Ceftiofur, Enrofloxacin, Florfenicol, Gentamicin, Penicillin, Tetracycline, Tilmicosin, and Trimethoprim-Sulfamethoxazole 
Journal of Clinical Microbiology  2001;39(12):4283-4287.
Quality control (QC) standards for the in vitro antimicrobial susceptibility testing of two fastidious veterinary pathogens, Actinobacillus pleuropneumoniae and Haemophilus somnus, were developed in a multilaboratory study according to procedures established by the National Committee for Clinical Laboratory Standards for broth microdilution and disk diffusion testing. The medium recommended for the broth microdilution testing is cation-adjusted Mueller-Hinton broth supplemented with 2% lysed horse blood, 2% yeast extract, and 2% supplement C. This medium has been designated veterinary fastidious medium. The medium recommended for the disk diffusion testing is chocolate Mueller-Hinton agar. The recommended QC organisms are A. pleuropneumoniae ATCC 27090 and H. somnus ATCC 700025. The QC MICs of ceftiofur, enrofloxacin, florfenicol, gentamicin, penicillin, tetracycline, tilmicosin, and trimethoprim-sulfamethoxazole were determined for each isolate, as were the zone size ranges. Of the results from the participating laboratories, 94.0% of the zone diameter results and 97.0% of the MIC results fell within the suggested QC ranges for all compounds. These QC guidelines should allow greater accuracy in interpreting results when testing these antimicrobial agents against fastidious pathogens.
PMCID: PMC88537  PMID: 11724833
19.  Pharmacokinetic evaluation of ceftiofur in serum, tissue chamber fluid and bronchial secretions from healthy beef-bred calves. 
Ceftiofur is a new broad spectrum cephalosporin marketed for the treatment of acute bovine respiratory disease. In this investigation ceftiofur was administered by intramuscular injection, at 24 h intervals, to healthy beef-bred calves for four days at dosages of 2.2 and 4.4 mg/kg of body weight, with 4 wk intervals between dosing regimens. Serum, tissue chamber fluid (TCF), and bronchial secretion (BS) concentrations of ceftiofur were measured by microbiological assay after the first and fourth dose of each dosing regimen. Peak serum concentrations (Cmax) of 8.8 micrograms/mL and 17.3 micrograms/mL were obtained approximately 2 h (Tmax), the time of mean peak concentration) after single injections of 2.2 mg/kg and 4.4 mg/kg, respectively. The Cmax was increased approximately twofold following multiple doses of 2.2 mg/kg (Cmax = 13.1 micrograms/mL) and 4.4 mg/kg (Cmax = 24.1 micrograms/mL). Ceftiofur accumulated slowly into TCF and peak concentrations were found to be approximately 14% of those observed in serum after the first dose and approximately 24% after multiple dosing. Concentrations of ceftiofur in BS were obtained rapidly with peak concentrations reaching 45% of the serum Cmax after the first dose. After multiple dosing the Cmax for BS was approximately 25% of the serum Cmax. This study found that both the 2.2 mg/kg and 4.4 mg/kg dosing regimens resulted in continuous serum, TCF and BS concentrations of ceftiofur that exceeded the minimal concentration required to inhibit the bacteria most frequently isolated from calves with acute bovine respiratory disease.
PMCID: PMC1263555  PMID: 1477795
20.  Knowledge of symptoms suggesting malignant disease amongst general practice patients 
A questionnaire completed by 119 general practice patients showed that patients often do not appreciate the importance of certain cancer symptoms. Patients' failure to respond to certain symptoms may be an important reason why they delay in seeking help.
PMCID: PMC1972265  PMID: 7086748
21.  Substance dependent American Indian veterans: a national evaluation. 
Public Health Reports  1994;109(2):235-242.
Demographic, clinical, and treatment episode characteristics of 3,087 American Indian veterans discharged from Department of Veterans Affairs hospitals in fiscal year 1991 were examined. Substance use disorders were diagnosed in 46.3 percent of discharged American Indians, compared with 23.4 percent of discharged veterans overall. More than 97 percent of American Indian substance use diagnoses were for alcohol dependence, while rates of other drug use disorders were low. Substance dependent American Indians were younger, and more likely to be male and unmarried, than nondependent American Indians. Psychiatric disorders, particularly personality disorders, depression, and posttraumatic stress disorders, were more prevalent among American Indians diagnosed with substance use disorders, than among nondependent American Indians. American Indians with substance use disorders were similar demographically to the general population of substance dependent veterans. Rates of diagnosed psychiatric disorders and drug dependencies other than alcohol were lower among American Indians receiving substance (alcohol or drug) use diagnoses than among the general population of substance dependent veterans. Rates of rehospitalization following discharge were higher in substance-abusing American Indian veterans than among their counterparts. Potential explanations for these findings are discussed.
PMCID: PMC1403480  PMID: 8153275
22.  Medical practice guidelines. 
Western Journal of Medicine  1994;161(1):39-44.
Prescriptive standards of clinical conduct--practice guidelines--have proliferated throughout medicine over the past decade. Practicing physicians are confronted with a plethora of guidelines developed for different purposes by a diverse body of public and private organizations. We review factors contributing to the growth of guidelines, their desirable features, and consequences, legal and otherwise, of implementing guidelines. Few studies have examined whether, and under what conditions, guidelines are effective in changing physicians' practices and patients' health. Nonetheless, expectations for guidelines remain high because they are one of the only instruments of health care reform that promises to improve the quality of care while reducing overall health care costs. Efforts to develop guidelines are likely to continue unabated for the foreseeable future. Additional research comparing different methods of developing and disseminating guidelines is needed.
PMCID: PMC1011365  PMID: 7941505
23.  Pulmonary immunity in calves following stimulation of the gut-associated lymphatic tissue by bacterial exotoxin. 
Antibodies in serum and pulmonary lavage fluids were measured in calves following stimulation of the gut-associated lymphatic tissue (GALT) by inoculation of crude leukotoxin of Pasteurella haemolytica into the duodenum through a surgically placed catheter. Nine calves free of P. haemolytica were divided into two groups. Group 1 received an intraduodenal (ID) inoculation of leukotoxin and group 2 received an ID inoculation of phosphate buffered saline. Serum and pulmonary lavage fluids were collected weekly and assayed for antibodies specific to P. haemolytica including immunoglobulin (Ig)G, leukotoxin neutralizing antibodies (LNA), and IgA (lavage fluids only). The multiplicative increase (over baseline) in each class of antibody titer following ID inoculation of leukotoxin, the composite geometric mean increase of all antibodies together, and the composite number of the five antibody titers which increased at least fourfold were computed. Results showed that the geometric mean of each antibody titer and the two composite indices was higher in the GALT-primed groups than in the sham-primed group. The differences were statistically significant (p less than 0.05) for serum IgG and for the two composite indices. This experiment demonstrates for the first time that GALT stimulation by bacterial exotoxins results in increased pulmonary antibody levels in calves.
PMCID: PMC1263522  PMID: 1591657
24.  Characterization of Gardnerella vaginalis and G. vaginalis-like organisms from the reproductive tract of the mare. 
Journal of Clinical Microbiology  1991;29(6):1157-1161.
Gardnerella vaginalis has been isolated from women with bacterial vaginosis, from the genital tracts of asymptomatic women, and from several other infected body sites in humans. However, until recently, it has not been isolated from any other animal species. Between June 1988 and October 1989, 31 isolates identified as G. vaginalis and 70 isolates identified as G. vaginalis-like organisms have been recovered from the genital tracts of 93 mares from Michigan and Ohio. Identification was based on biochemical reactions, hemolysis on media containing blood from various animal sources, and susceptibility to select antimicrobial agents. This report details the characterization of G. vaginalis and G. vaginalis-like organism isolates obtained from the reproductive tracts of these mares and compares the equine isolates with human isolates.
PMCID: PMC269962  PMID: 1864934
25.  Hematological changes in calves exposed to a mixture of lipopolysaccharide and crude leukotoxin of Pasteurella haemolytica. 
The purpose of this investigation was to determine if culture supernatants of Pasteurella haemolytica containing crude leukotoxin and lipopolysaccharide (CLCL) causes disseminated intravascular coagulopathy (DIC) when injected into calves. The effect of intraduodenal (ID) exposure followed by a subsequent subcutaneous (SC) inoculation of either heat-treated or untreated CLCL was evaluated. The relative contribution of the crude leukotoxin and lipopolysaccharide (LPS) to the virulence of P. haemolytica was evaluated. One group of calves received an ID inoculation of CLCL followed two weeks later by a SC inoculation of CLCL; one group received an ID inoculation of tissue culture medium followed two weeks later by a SC inoculation of CLCL; and a third group received an ID inoculation of CLCL followed two weeks later by a SC inoculation of heat-treated CLCL. Hematological parameters used to evaluate DIC included white cell count, platelet count, neutrophil number, fibrinogen, fibrin degradation products, one stage prothrombin time (OSPT), activated partial thromboplastin time, body temperature and clinical signs. Each parameter was measured in calves at 0, 2, 4, 6, 12 and 24 h following the SC inoculation of CLCL. Each group had significant changes over time in all parameters except body temperature. Calves that received a SC inoculation of heat-treated CLCL had smaller changes in all parameters except OSPT compared to the other groups. Results suggest that the LPS and leukotoxin of P. haemolytica exert additive effects on the coagulation cascade and number of peripheral leukocytes, and that the ID inoculation of CLCL does not affect the response of calves to a SC inoculation of toxin.
PMCID: PMC1255686  PMID: 2249175

Results 1-25 (29)