Effective surface area on rough substrates for bacterial adhesion is examined by analyzing the solid fraction of surfaces, where medium in contact with surface.

To overcome problems associated with application of PCR to clinical samples, we have combined a short cultivation procedure Salmonella-specific PCR-hybridization assay specifically identify Salmonella serovars from samples various animal species. The technique was investigated by using fecal seeded known numbers organisms and cultivated for different lengths time in assorted selective nonselective enrichment media. ability amplify DNA product (457-bp sequence covering the invE invA genes)...

Presumptive identification of Escherichia coli O157:H7 is possible in an individual, nonmultiplexed PCR if the reaction targets enterohemorrhagic E. (EHEC) eaeA gene. In this report, we describe development and evaluation sensitivity specificity a PCR-based 5' nuclease assay for presumptively detecting DNA. The eaeA-based system was sufficient to correctly identify all strains evaluated, mirroring previously described primers. SZ-primed, eaeA-targeted detection capable rapid, semiautomated,...

Abstract Objective —To evaluate, under field conditions, the effects of a commercial porcine circovirus type 2 (PCV2) vaccine on mortality rate and growth performance in herd infected with PCV2 that had history disease. Design —Randomized controlled clinical trial. Animals —485 commercial, cross-bred, growing pigs. Procedures —Prior to weaning, pigs were randomly assigned within litter vaccination or unvaccinated control group. Pigs group given at weaning 3 weeks later. Mortality was...

The bacterial effector proteins SseK and NleB glycosylate host on arginine residues, leading to reduced NF-κB-dependent responses infection. Salmonella SseK1 SseK2 are E. coli NleB1 orthologs that behave as NleB1-like GTs, although they differ in protein substrate specificity. Here we report these enzymes retaining glycosyltransferases composed of a helix-loop-helix (HLH) domain, lid catalytic domain. A conserved HEN motif (His-Glu-Asn) the active site is important for enzyme catalysis...

ABSTRACT We have developed a rapid procedure for the detection of virulent Yersinia enterocolitica in ground pork by combining previously described PCR with fluorescent dye technologies. The method, known as fluorogenic 5′ nuclease assay (TaqMan), produces results measuring fluorescence produced during amplification, requiring no post-PCR processing. specificity chromosomal yst gene-based was tested 28 bacterial isolates that included 7 pathogenic and nonpathogenic serotypes Y. , other...

Many Gram-negative bacterial pathogens interact with mammalian cells by using type III secretion systems (T3SS) to inject virulence proteins into host cells. A subset of these injected protein 'effectors' are enzymes that inhibit the function catalyzing addition unusual post-translational modifications. The E. coli and Citrobacter rodentium NleB effectors, as well Salmonella enterica SseK effectors glycosyltransferases modify substrates N-acetyl glucosamine (GlcNAc) on arginine residues....

Abstract Liver abscesses in feedlot cattle are polymicrobial infections. Culture-based studies have identified Fusobacterium necrophorum as the primary causative agent, but a number of other bacterial species frequently isolated. The incidence liver is highly variable and affected by factors, including type. Holstein steers raised for beef production higher than crossbred cattle. Tylosin commonly used antimicrobial feed additive to reduce abscesses. objective this study was utilize 16S...

Outer membrane vesicles (OMVs) are promising vaccine components because they combine antigen and adjuvant in a single formulation. Detoxified Salmonella enterica strains that express penta-acylated lipid A retain OMV immunogenicity but with reduced reactogenicity. We have previously shown recombinant form of the enterotoxigenic Escherichia coli (ETEC) seventeen kilodalton protein (Skp) protects mice pulmonary challenge model, when fused to glutathione-S-transferase (GST) epitope combined...

Type III secretion system (T3SS) effector proteins are primarily recognized for binding host to subvert immune response during infection. Besides their known target proteins, several T3SS effectors also interact with endogenous bacterial proteins. Here we demonstrate that the Salmonella glycosyltransferase SseK1 glycosylates two-component regulator OmpR on two arginine residues, R15 and R122. Arg-glycosylation of results in reduced expression ompF, a major outer membrane porin gene....

Culture methods for detecting virulent Yersinia enterocolitica require selective enrichment and a series of confirmatory tests that are time-consuming, costly, laborious. The objective this study was to evaluate fluorogenic 5′-nuclease assay the enterotoxin yst gene Y. in pure cultures, inoculated ground pork samples, naturally contaminated food samples. These results were then compared with "gold standard" recommended by U.S. Food Drug Administration Bacteriological Analytical Manual...

Enterotoxigenic Escherichia coli (ETEC) causes diarrheal disease. Antigenic and structural heterogeneity among ETEC colonization factors has complicated vaccine development efforts. Identifying characterizing conserved antigens that induce protective immunity is therefore of interest. We previously characterized three proteins (MipA, Skp, ETEC_2479) protected mice in an intranasal challenge model after vaccination. However, these are not only multiple isolates, but also commensal bacteria....

A rapid and sensitive cultivation PCR-hybridization procedure for the detection identification of Salmonella typhimurium was evaluated over a 42-day period with eight experimentally infected beagles. Rectal swabs were taken at several times postinfection, inoculated into selenite-cystine broth, plated onto Hektoen-Enteric Enteric agar immediately after incubation 4 24 h. PCRs hybridizations also conducted each sample, results compared those standard culture techniques to evaluate efficiency...

An assay to identify tissue culture cells infected with bovine respiratory syncytial virus (BRSV) that utilizes reverse transcription (RT), the polymerase chain reaction (PCR), and a synthetic oligonucleotide hybridization probe has been developed. The RT-PCR uses BRSV-specific negative-sense primer synthesize cDNA from BRSV fusion protein mRNA template another (positive sense) upstream for PCR amplification. In presence of templates isolates originating locations throughout United States,...

Bats have been implicated as potential carriers of Leptospira a result surveys, mostly in Australia and South America. We measured the prevalence pathogenic leptospires kidneys bats from Kansas Nebraska. From 7 August 2012 to 21 2012, we extracted DNA 98 big brown (Eptesicus fuscus) submitted found negative for rabies. The was processed two-step, seminested PCR assay with dual-labeled Taqman probe specific leptospires. As control, used saprophytic leptospire (Leptospira biflexa Patoc) and,...

OBJECTIVE To use variable-number tandem-repeat (VNTR) analysis to determine the infecting serovar and strain for leptospiral DNA isolated from canine urine samples confirmed through PCR testing contain pathogenic leptospires evaluate sensitivity specificity of microscopic agglutination (MAT) identifying serogroup. DESIGN Diagnostic survey test evaluation. SAMPLE Leptospiral 98 dogs have in their urine. PROCEDURES VNTR isolates was performed identify by primer pairs loci 4, 7, 10, Lb5....

The Salmonella enterica SseK1 protein is a type three secretion system effector that glycosylates host proteins during infection on specific arginine residues with N-acetyl glucosamine (GlcNAc). also Arg-glycosylates endogenous bacterial and we thus hypothesized activities might be integrated regulating the intrabacterial abundance of UPD-GlcNAc, sugar-nucleotide donor used by this effector. After searching for new substrates, found in dual repressor-activator NagC, leading to increased...

We have developed a rapid and sensitive assay for the detection of Salmonella serovars in veterinary clinical specimens. This method utilizes short cultivation period followed by PCR. For amplified product, an enzyme-linked immunosorbent (ELISA)-based oligonucleotide ligation (OLA) was used. In this study, PCR-OLA technique compared with conventional culture membrane hybridization bacteria. evaluating non-Salmonella strains bacteria, A490 readings 51 strains, representing 28 serovars, were...

Eperythrozoon suis is an extracellular red blood cell parasite that causes icteroanemia in acutely ill pigs and a variety of syndromes chronically infected pigs. Current techniques to detect E. infection are limited by variability parasitemias antibody responses animals. The polymerase chain reaction (PCR) was investigated determine its potential as means detecting With DNA samples extracted from either purified organisms or suis-infected pig blood, PCR produced amplification product 492...

Objectives: To compare the effects of porcine circovirus type 2 (PCV2) vaccination on growth rate, backfat depth, and loin depth pigs in a high-health herd which contained different genetic lines. Materials methods: A total 454 (20.6 ± 1.98 days age; 6.1 1.27 kg body weight) were used 130-day randomized controlled field trial. Genetic designations A×A (Duroc line), B×B (synthetic White Pietrain A×B, B×A. Pigs randomly assigned to treatments (Vaccinated or nonvaccinated Control) within litter...

In reverse transcription-polymerase chain reactions (RT-PCR) and DNA hybridizations using primers an oligonucleotide probe to the fusion (F) protein mRNA of bovine respiratory syncytial virus (BRSV), all BRSV isolates a goat isolate could be distinguished from prototype human viruses (HRSV) ovine (sheep bighorn sheep) (RSV). However, RT-PCR amplifications with sequences HRSV F resulted in amplified products ≍ 243 bp if templates subgroup A strains were present slightly larger B strains. No...

Currently, methods for recovering and identifying Escherichia coli O157:H7 from cattle feces are inconsistent hindered by their inability to specifically rapidly detect small numbers of organisms this complex highly variable matrix. A standard approach isolating characterizing E. was compared with a polymerase chain reaction (PCR)-based 5′ nuclease assay specific that included secondary enrichment step. The PCR-based method proved better indicator the presence organism than culture...

Achieving cross-protective efficacy against multiple bacterial strains or serotypes is an important goal of vaccine design. Enterotoxigenic Escherichia coli (ETEC) cause diarrheal disease in underdeveloped nations. We have been interested identifying and characterizing ETEC antigens that generate protective immune responses independent colonization factor (CF) expression. Our previous studies used proteomics to identify the MipA, Skp, ETEC_2479 proteins as effective protecting mice from...