Glutamate imaging (GluCEST) reveals lower brain GluCEST contrast in patients on the psychosis spectrum.

Psychosis commonly develops in adolescence or early adulthood. Youths at clinical high risk (CHR) for psychosis exhibit similar, subtle symptoms to those with schizophrenia (SZ). Malfunctioning neurotransmitter systems, such as glutamate, are implicated in the disease progression of psychosis. Yet, in vivo imaging techniques for measuring glutamate across the cortex are limited. Here, we use a novel 7 Tesla MRI glutamate imaging technique (GluCEST) to estimate changes in glutamate levels across cortical and subcortical regions in young healthy individuals and ones on the psychosis spectrum. Individuals on the psychosis spectrum (PS; n=19) and healthy young individuals (HC; n=17) underwent MRI imaging at 3 and 7 T. At 7 T, a single slice GluCEST technique was used to estimate in vivo glutamate. GluCEST contrast was compared within and across the subcortex, frontal, parietal and occipital lobes. Subcortical (χ2 (1)=4.65, P=0.031) and lobular (χ2 (1)=5.17, P=0.023) GluCEST contrast levels were lower in PS compared with HC. Abnormal GluCEST contrast levels were evident in both CHR (n=14) and SZ (n=5) subjects, and correlated differentially, across regions, with clinical symptoms. Our findings describe a pattern of abnormal brain neurochemistry early in the course of psychosis. Specifically, CHR and young SZ exhibit diffuse abnormalities in GluCEST contrast attributable to a major contribution from glutamate. We suggest that neurochemical profiles of GluCEST contrast across cortex and subcortex may be considered markers of early psychosis. GluCEST methodology thus shows promise to further elucidate the progression of the psychosis disease state.

An experimental platform for investigation of the Zeeman effect in strong magnetic fields.

An experimental platform is developed for the investigation of the Zeeman effect in strong magnetic fields. Mega-Gauss magnetic fields are generated by a 1 MA Zebra pulsed power machine using metal rod loads. A gas jet or CH oil on the load is the source of hydrogen. Excited hydrogen atoms are backlit by black body radiation from the rod load. Hydrogen absorption spectra are recorded with a grating spectrometer and intensified gated CCD camera. The experimental platform enables the observation of the quadratic Zeeman effect in hydrogen gas jets using the spectral shift of the central line in the Zeeman triplet. Other gases can be studied using the gas jet method.

Quadratic Zeeman effect in hydrogen at 2-3 MG magnetic fields.

The Zeeman effect is used for measurement of magnetic fields in astrophysical and laboratory plasmas. Magnetic fields in atmospheres of magnetic white dwarf stars are in the range 40 kG-1 GG. The quadratic Zeeman effect results in the additional split and shift of lines for magnetic fields >2 MG. Hydrogen Balmer lines were studied in magnetic fields delivered by a 1 MA pulse power generator. The magnetic field was generated by rod loads 0.8-1 mm in diameter. A droplet of CH oil on the load center was the source of hydrogen. A low ionized oil layer was backlit by blackbody emission from the rod with a temperature of 0.5-0.6 eV. Zeeman splitting of H-alpha and H-beta absorption lines were with a grating spectrometer. A spectral shift of the central component of the triplet indicated the quadratic Zeeman effect in hydrogen lines.

DNA identification and characterization of Campylobacter jejuni and Campylobacter coli isolated from caecal samples of chickens in Grenada.

AIMS: To speciate Campylobacter strains from the caeca of chickens in Grenada using PCR and to evaluate DNA-based typing methods for the characterization of these isolates. METHODS AND RESULTS: Isolates were speciated with two multiplex PCR assays and were typed with flaA-RFLP, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results confirmed that Campylobacter coli strains were more predominant than Campylobacter jejuni strains. From 56 isolates, 18 were misidentified using biochemical tests. PFGE typing gave the highest discriminatory power among the methods used (Simpson's index of diversity, D=0.9061). However, the combination of flaA-RFLP, PFGE and MLST results gave the highest discrimination for subtyping of these isolates (D=0.9857). A band position tolerance of 4% in BioNumerics was the most appropriate for the analysis of this database. MLST profiles were generally concordant with PFGE and/or flaA-RFLP types. Several isolates exhibited new MLST sequence types (STs), and 43 of the 49 Camp. coli strains belonged to the ST-828 clonal complex. CONCLUSIONS: Campylobacter coli was the most prevalent species isolated from broilers and layers in Grenada, and a combination of restriction and sequence methods was most appropriate for the typing of Camp. coli isolates. Campylobacter coli STs clustered with described poultry-associated Camp. coli STs by phylogenetic analysis. SIGNIFICANCE AND IMPACT OF THE STUDY: Further studies to understand the predominance of Camp. coli within Campylobacter spp. from chickens in Grenada may help elucidate the epidemiology of these pathogens in chickens.

Observations on production of hemolysin, heat-labile enterotoxin and antimicrobial drug resistance among enterotoxigenic Escherichia coli from pigs.

A total of 52 isolates of Escherichia coli belonging to enterotoxigenic serotypes from piglets with diarrhea were examined for hemolysis, production of cholera-like enterotoxin (LT) and susceptibility to 10 antimicrobial drugs. A strong association between production of LT and hemolysis was seen. Ninety percent of 29 hemolylic isolates were LT+ whereas 100% of 23 nonhemolytic isolates were LT- in a commercial latex agglutination assay. Antimicrobial susceptibility tests employing disc diffusion showed that resistance to trimethoprim-sulfamethoxazole (TMS), neomycin and tetracycline was significantly less among LT+ isolates compared to LT- ones. Enrofloxacin was the only antimicrobial drug to which all the 52 isolates were susceptible.

Characterization of Salmonella isolates from beef cattle, broiler chickens and human sources on Prince Edward Island.

Non-typhoid Salmonella serovars remain a potential threat to human health, and beef cattle and broiler chickens are possible sources of these organisms on Prince Edward Island (PEI). In this study, the ceca of beef cattle belonging to fasted and non-fasted groups, and broiler chickens were examined for Salmonella at the time of slaughter. The characteristics of the isolates, including antimicrobial resistance patterns and virulence genes, were studied along with the isolates obtained from cases of human salmonellosis on PEI during the study period (1996-97). The prevalence of Salmonella in beef cattle was 4.6% (11/240). The rate was significantly higher in fasted cattle (7.46%), than in non-fasted cattle (0.94%). The prevalence rate in chickens was 32.5% (39/120). In beef cattle, Salmonella typhimurium phage type (PT) or definitive type (DT) 104 which was resistant to ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline, was the most predominant type (64%). In chickens, S. heidelberg, with resistance to gentamicin, streptomycin and sulfisoxazole, predominated. Of 26 isolates from humans, the most common serovar was S. typhimurium, including a multidrug-resistant strain of DT104. Examination by PCR revealed presence of the virulence gene invA in all serovars, and the spvC gene in all S. typhimurium isolates, of both beef cattle and human origin. Among the other serovars the latter gene was found in 7 human isolates, but in none of the chicken or beef isolates. All but 3 of the spvC-positive isolates possessed a 90 kilobasepair (kbp) plasmid suggesting that the 3 isolates had the spvC gene on their chromosome. These findings were confirmed by plasmid DNA isolation using 3 different protocols and by sequence analysis of the spvC-PCR product.

Prevalence and characteristics of shiga toxin-producing Escherichia coli in beef cattle slaughtered on Prince Edward Island.

Fecal swabs obtained from a random sample of 1,000 beef slaughter steers and heifers from 123 Prince Edward Island (P.E.I.) farms were examined for the presence of Shiga toxin-producing Escherichia coli (STEC) using a Vero cell assay (VCA). Multiple isolates from each positive sample were tested similarly. VCA-positive isolates were confirmed as E. coli biochemically, tested for drug resistance, serotyped, and tested by polymerase chain reaction (PCR). Animals were classified as positive when an isolate was positive on VCA and the presence of the gene responsible for toxin production was confirmed by PCR. The prevalence of STEC in beef slaughter steers and heifers on P.E.I. was 4% (40 of 1,000). The total number of isolates was 43, and these comprised 26 serotypes, including 13 isolates belonging to 6 serotypes known to be associated with human illness. The most frequently isolated STEC serotype was E. coli O157 (5 isolates out of 43). Of the five E. coli O157 isolates, four were E. coli O157:H7, a serious human pathogen. The majority of STEC isolates, including all O157:H7, isolates, were susceptible to 16 commonly used antimicrobial drugs. According to PCR, 65% of the STEC isolates had the gene for Stx1. Four of these isolates, including two O157:H7, had genes for Shiga toxin (Stx)1 and Stx2.

An investigation of bacterial causes of arthritis in slaughter hogs.

Joints from 153 arthritic and 80 normal slaughter hogs were examined by culture for presence of bacteria. Although none of the normal joints yielded bacteria, 37% of the disease joints were positive for bacterial growth. Of 67 bacterial isolates obtained, 45% were Erysipelothrix rhusiopathiae. Occurrence of other bacteria in order of their frequency was Streptococcus suis (16%), Actinomyces pyogenes (10%), Mycoplasma spp. including 3 M. hyorhinis isolates (7%), staphylococci (7%), Streptococcus spp. (6%), and organisms of uncertain significance (7%).

Comparison of two pre-surgical skin preparation techniques.

A one-step iodophor skin preparation solution was compared to chlorhexidine gluconate application as a pre-operative skin preparation method in 100 animals undergoing elective ovariohysterectomy. Pre-operative and intra-operative skin cultures demonstrated no difference in antiseptic efficacy. No animal in the study demonstrated signs of systemic infection, and no adverse local effects from either antiseptic were seen. The iodophor solution evaluated (DuraPrep) is a safe and effective pre-operative skin preparation agent in small animal patients undergoing clean surgical procedures.

A cohort study of coagulase negative staphylococcal mastitis in selected dairy herds in Prince Edward Island.

The epidemiology and importance of coagulase negative staphylococcal (CNS) mastitis in Prince Edward Island had not been documented. To investigate this, a cohort of 84 cows at seven farms were quarter sampled eight times over a lactation, commencing with samples taken prior to drying off in the previous lactation. Thirteen species of CNS were isolated. The quarter prevalence of CNS mastitis varied from 4.8% to 6.4% in the first five months of lactation and increased to 14.2 to 16.6% in the last four months of lactation. The geometric mean somatic cell counts (SCC) for quarters infected with CNS and uninfected quarters were 90 x 10(3) and 64 x 10(3) respectively (difference significant at p > 0.005). The two month new infection risk of CNS was 9.0% while the two month elimination risk was 74.4%. Infection with CNS did not alter the risk of subsequent infection with Staphylococcus aureus. The results from this project support the classification of CNS as a minor pathogen in mastitis control programs.

Development of a specific biotinylated DNA probe for the detection of Renibacterium salmoninarum.

A specific DNA probe for the identification of Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD), was developed from one of 3 clones pRS47, pRS49, and pRS26 of 5.1 kb, 5.3 kb, and 11.3 kb, respectively. The biotinylated pRS47/BamHI insert probe was tested on 3 dilutions of DNA extracted from 3 strains of R. salmoninarum and from 1 strain each of Arthrobacter protophormiae, Aeromonas salmonicida, Corynebacterium aquaticum, Carnobacterium piscicola, Listonella anguillarum, Micrococcus luteus, Pseudomonas fluorescens, Vibrio ordalii, and Yersinia ruckeri. In a dot blot assay, this probe hybridized only with the DNA from the R. salmoninarum strains. When used on kidney samples from fish challenged with R. salmoninarum, the dot blot hybridization assay with the probe was found to be as sensitive as culture. In a fluorescent antibody test, samples that were negative in culture and dot blot hybridization showed no more than one fluorescing cell in 50 microscopic fields examined. This DNA probe, therefore, has the potential for use in the diagnosis of BKD of fish.

Isolation, serotypes, and virulence-associated properties of Yersinia enterocolitica from the tonsils of slaughter hogs.

The objectives of this study were to determine the carriage rate of Yersinia enterocolitica in the tonsils of slaughter hogs, and to characterize them with regard to phenotypic and virulence-associated properties. Of 202 pigs examined from an abattoir in Prince Edward Island, 85 were culture positive for Y. enterocolitica. Sixty-seven percent of isolates belonged to serotype O:3, and 20% were serotype O:5. All isolates produced urease and 95% of O:3 isolates showed virulence-associated characters of autoagglutination at 37 degrees C and lack of fermentation of esculin and salicin. All isolates were tested for crystal violet binding, calcium dependency, and virulence plasmids. Eight isolates (5 belonging to serotype O:3, 2 belonging to O:5,27, and 1 belonging to O:7,8) were tested in addition for the production of heat-stable enterotoxin (ST), and iron-chelating siderophores. Of the 57 O:3 isolates, 93% were positive for crystal violet binding and calcium dependency and 98% possessed a 40-45 MDa plasmid. Four of the 5 O:3 isolates tested for ST related to Escherichia coli STa in a commercial enzyme immunoassay were positive. Six of the 8 isolates belonging to 3 different serotypes produced large orange halos around the colonies on a chrome-azurol-s agar assay medium, for siderophores. Antimicrobial susceptibility tests of all 85 isolates against 16 drugs showed 100% susceptibility against 12 drugs, including trimethoprim-sulfamethoxazole and tetracycline.

A microbiological assay for determining sarafloxacin and oxolinic acid concentrations in Atlantic salmon plasma.

A microbiological assay was developed to quantify the concentrations of the quinolone antibiotics sarafloxacin and oxolinic acid in Atlantic salmon (Salmo salar) plasma. The assay was a modification of the Association of Official Analytical Chemists (AOAC) method for antibiotic concentrations in feeds using disk diffusion and a lawn of Vibrio anguillarum ATCC 19264 as the test organism. With these modifications, sensitivities of 0.04 microgram/mL for sarafloxacin and 0.125 micrograms/mL for oxolinic acid were obtained.

Toxigenicity of culture filtrates of Salmonella enteritidis isolates on three mammalian cell lines.

Culture filtrates of 28 Salmonella enteritidis isolates were tested for toxicity on Vero-, CHO-, and human foreskin fibroblast (HFF) cells. Cytopathic effects on HFF cells were extensive, and were observed even with some filtrates diluted 1:256. Vero cells showed effects with filtrates diluted up to 1:16, and CHO cells gave weak or no reaction. All isolates produced iron-binding siderophores as determined by reactions on chrome-azurol-S medium.

Detection of Campylobacter jejuni/C.coli infection in patients with Guillain-Barré syndrome by serology and culture.

Serum samples from 43 cases of Guillain-Barré Syndrome (GBS), 32 non-GBS neurology patients, and 35 healthy persons from a medical institute in Kerala, India, were tested for antibody levels against a Campylobacter jejuni strain Penner serotype 0:19 by agglutination and ELISA. Twenty-six percent of samples from GBS cases showed high antibody levels in all assays. Of 8 stool samples of new GBS cases examined by culture, 38% were positive for C. jejuni/C. col1. The results suggest that at least a quarter of GBS cases studied were associated with Campylobacter infection.

Prevalence, serovars and antimicrobial susceptibility of Salmonella spp. from wild and domestic green iguanas (Iguana iguana) in Grenada, West Indies.

Cloacal swabs from 62 green iguanas (Iguana iguana), including 47 wild and 15 domestic ones from five parishes of Grenada, were sampled during a 4-month period of January to April 2013 and examined by enrichment and selective culture for the presence of Salmonella spp. Fifty-five per cent of the animals were positive, and eight serovars of Salmonella were isolated. The most common serovar was Rubislaw (58.8%), a serovar found recently in many cane toads in Grenada, followed by Oranienburg (14.7%), a serovar that has been causing serious human disease outbreaks in Japan. Serovar IV:48:g,z51 :- (formerly, S. Marina) highly invasive and known for serious infections in children in the United States, constituted 11.8% of the isolates, all of them being from domestic green iguanas. Salmonella Newport, a serovar recently found in a blue land crab in Grenada, comprised 11.8% of the isolates from the green iguanas. The remaining four less frequent serovars included S. Javiana and S. Glostrup. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin-clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, nalidixic acid, streptomycin, tetracycline and trimethoprim-sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, mainly to streptomycin, tetracycline and cefotaxime. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars from wild and domestic green iguanas in Grenada, West Indies.

Prevalence of Salmonella spp. in cane toads (Bufo marinus) from Grenada, West Indies, and their antimicrobial susceptibility.

Cloacal swabs and caecal contents sampled from 58 cane toads (Bufo marinus) in St George's parish, Grenada, during a 7-month period in 2011 were examined by an enrichment and selective culture method for presence of Salmonella spp. Twenty-four (41%) toads were positive for Salmonella spp. of which eight were Salmonella enterica serovar Javiana, and eight were S. enterica serovar Rubislaw. The other serovars were as follows: Montevideo, 6; Arechavaleta, 1; and serovar: IV:43:-:-, 1. The high frequency of isolation of serovar Javiana, an emerging human pathogen associated with several outbreaks in the recent years in the eastern United States, suggests a possible role for cane toads in transmission of this serovar. Although S. Rubislaw has been isolated from lizards, bats and cases of some human infections, there is no report of its carriage by cane toads, and in such high frequency. The rate of carriage of S. Montevideo, a cause for human foodborne outbreaks around the world was also over 10% in the 58 toads sampled in this study. The antimicrobial drug susceptibility tests against amoxicillin-clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim-sulfamethoxazole showed that drug resistance is minimal and is of little concern. Antimicrobial resistance was limited to ampicillin and amoxicillin-clavulanic acid in one isolate of S. Javiana and one isolate of S. Rubislaw. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars not identified previously in cane toads in Grenada, West Indies.

Serologic detection of antibodies to Brucella spp. using a commercial ELISA in cattle in Grenada, West Indies.

Bovine brucellosis, caused mainly by Brucella abortus, a zoonotic bacterium, has been reported from many areas of the world, including Central and South America, and the Caribbean island state of Trinidad and Tobago. Although brucellosis has been eradicated from domestic cattle in Canada it still exists in one or two herds in the United States. Serological tests are important in estimating prevalence of Brucella exposure in order to target eradication programmes. In this study, serum samples from 150 cattle were tested using a commercial competitive enzyme linked immunosorbent assay (SVANOVIR®Brucella-Ab C-ELISA) which detects antibodies to both B. abortus and Brucella melitensis. All cattle tested were greater than 6 months old and were unvaccinated. Sampled cattle were from 35 herds representing animals from all 6 parishes of Grenada. Nine of the 150 animals (6%) were positive for antibodies to B. abortus and/or melitensis by the C-ELISA. Of the 35 herds, 7 (20%) had C-ELISA- positive animals. Three of the 6 parishes contained positive herds. Based on the high sensitivity (98%) and specificity (99.7%) of the C-ELISA, these results strongly indicate the presence of cattle exposed to B. abortus and/or melitensis in Grenada.