Complete genome sequences of strains ASE2 and ASE4 generated by PacBio sequencing.

strains ASE2 and ASE4 were isolated from the soil of a lettuce farm in Salinas, California, in 2020. Their complete genome sequences were generated by the PacBio Sequel II platform. They both consist of a single circular chromosome without plasmids.

Biocontrol of Escherichia coli O157:H7 by Enterobacter asburiae AEB30 on intact cantaloupe melons.

Escherichia coli O157:H7 causes >73,000 foodborne illnesses in the United States annually, many of which have been associated with fresh ready-to-eat produce including cantaloupe melons. In this study, we created a produce-associated bacterial (PAB) library containing >7500 isolates and screened them for the ability to inhibit the growth of E. coli O157:H7 using an in vitro fluorescence-based growth assay.

Complete genome sequence of ASE1 generated by PacBio sequencing.

We report the complete genome sequence of ASE1 generated by the PacBio Sequel II platform. This bacterium was isolated from the soil of a lettuce farm in Salinas, CA, USA, in 2020. The genome consists of a single circular chromosome of 5,021,820 bp with a 52.

Use of Pantoea agglomerans ASB05 as a biocontrol agent to inhibit the growth of Salmonella enterica on intact cantaloupe melons.

Aims: To identify biocontrol agents to prevent the growth of Salmonella serotype Enterica on cantaloupe melons during the pre- and postharvest periods.

Methods And Results: We created a produce-associated bacterial library containing 8736 isolates and screened it using an in-vitro fluorescence inhibition assay to identify bacteria that inhibit the growth of S. Enterica.

Sensitive and selective detection of peanut allergen Ara h 1 by ELISA and lateral flow immunoassay.

The Ara h1 protein is a peanut allergen and it provides a useful biomarker for the detection of peanut protein. In this manuscript, we describe the generation of monoclonal antibodies (MAbs) against the Ara h1 protein and their development into sensitive and selective immunoassays for peanut detection. Our enzyme-linked immunosorbent assay (sELISA) detects a peanut meal standard with a sensitivity of 10 ng/mL and 500 ng/mL by lateral flow immunoassay (LFIA).

Role of soil in the regulation of human and plant pathogens: soils' contributions to people.

Soil and soil biodiversity play critical roles in Nature's Contributions to People (NCP) # 10, defined as Nature's ability to regulate direct detrimental effects on humans, and on human-important plants and animals, through the control or regulation of particular organisms considered to be harmful. We provide an overview of pathogens in soil, focusing on human and crop pathogens, and discuss general strategies, and examples, of how soils' extraordinarily diverse microbial communities regulate soil-borne pathogens. We review the ecological principles underpinning the regulation of soil pathogens, as well as relationships between pathogen suppression and soil health.

Complete Genome Sequence of Enterobacter asburiae Strain AEB30, Determined Using Illumina and PacBio Sequencing.

The complete genome sequence of Enterobacter asburiae strain AEB30 is presented. The strain was isolated from store-bought ginger in Albany, CA, in 2016.

Complete Genome Sequence of Pantoea agglomerans ASB05 Using Illumina and PacBio Sequencing.

We present the complete genome sequence of Pantoea agglomerans ASB05 and three associated plasmids, generated using a combination of the Illumina and PacBio platforms. ASB05 was isolated from fresh cherries purchased in Albany, CA, in 2016.

A rapid and sensitive lateral flow immunoassay (LFIA) for the detection of gluten in foods.

The gluten protein found in a variety of cereal grains is a food allergen that can elicit a spectrum of immuno-inflammatory responses in people. Consumer awareness has prompted changes in food labeling requirements, expanded gluten-free food product availability and increased demand for effective gluten testing methodologies. To meet the challenges associated with gluten testing from diverse and complex foods we developed a lateral flow immunoassay (LFIA) using a pair of novel gliadin monoclonal antibodies (MAbs).

ALB65 Inhibits the Growth of on Cantaloupe Melons.

is a foodborne pathogen that causes high rates of hospitalization and mortality in people infected. Contamination of fresh, ready to eat produce by this pathogen is especially troubling because of the ability of this bacterium to grow on produce under refrigeration temperatures. In this study, we created a library of over 8,000 plant phyllosphere-associated bacteria and screened them for the ability to inhibit the growth of in an fluorescence-based assay.

Complete Genomic Sequences of Three Salmonella enterica subsp. Serovar Muenchen Strains from an Orchard in San Joaquin County, California.

We present here the complete genome sequences of three subsp. serovar Muenchen strains, LG24, LG25, and LG26. All three strains were isolated from almond drupes grown in an orchard in San Joaquin County, California, in 2016.

Use of Phyllosphere-Associated Lactic Acid Bacteria as Biocontrol Agents To Reduce Serovar Poona Growth on Cantaloupe Melons.

Foodborne illness associated with fresh, ready-to-eat produce continues to be a significant challenge to public health. In this study, we created a phyllosphere-associated lactic acid bacteria (PLAB) library and screened it via a high-throughput in vitro fluorescent assay to identify bacteria capable of inhibiting the growth of the pathogenic bacterium . One isolate, 14B4, inhibited the growth of by >45-fold in vitro; it was able to grow and persist on the surfaces of cantaloupe melons at both ambient (25°C) and refrigerator (5°C) temperatures.

Rapid Detection of Staphylococcal Enterotoxin-B by Lateral Flow Assay.

A cohort of monoclonal antibodies (mAbs) were generated against Staphylococcal enterotoxin-B (SEB) and selected by double sandwich enzyme-linked immunosorbent assay (ELISA) for solution capture of the toxin. Clonal hybridoma cell lines were established and a pair of anti-SEB mAbs selected for the development of a sandwich ELISA. Immobilized 3D6 mAb (IgG1, kappa) when paired with 4C9 mAb (IgG1, kappa) conjugated to horseradish peroxidase generates a typical dose-response curve with an EC of 24.

Dosage-dependent effects of monensin on the rumen microbiota of lactating dairy cattle.

We examined the dose-dependent effects of feeding lactating dairy cows a standard diet supplemented with monensin at 175, 368, or 518 mg cow day on the rumen microbiota. For each dosage, 3 animals were randomly assigned into groups and fed the same basal total mixed ration diet supplemented with monensin, at the respective dose. After 20 days, rumen samples were taken and the effect on the microbiota was examined by 16S rRNA gene sequence analysis and qPCR.

Complete Genome Sequence of Lactococcus lactis subsp Strain 14B4, Which Inhibits the Growth of Salmonella enterica Serotype Poona .

We present here the complete genome sequence of Lactococcus lactis strain 14B4, isolated from almond drupes in northern California. This strain was observed to inhibit the growth of Salmonella enterica serotype Poona strain RM3363 .

Complete Genome Sequences of Three Bacillus amyloliquefaciens Strains That Inhibit the Growth of Listeria monocytogenes .

Here, we report the complete genome sequences of three strains isolated from alfalfa, almond drupes, and grapes that inhibited the growth of strain 2011L-2857 We also report multiple gene clusters encoding secondary metabolites that may be responsible for the growth inhibition of .

Identification and verification of hybridoma-derived monoclonal antibody variable region sequences using recombinant DNA technology and mass spectrometry.

Antibody engineering requires the identification of antigen binding domains or variable regions (VR) unique to each antibody. It is the VR that define the unique antigen binding properties and proper sequence identification is essential for functional evaluation and performance of recombinant antibodies (rAb). This determination can be achieved by sequence analysis of immunoglobulin (Ig) transcripts obtained from a monoclonal antibody (MAb) producing hybridoma and subsequent expression of a rAb.

Affinity Purification of Antibodies.

Antibodies are provided in a variety of formats that include antiserum, hybridoma culture supernatant, or ascites. They can all be used successfully in crude form for the detection of target antigens by immunoassay. However, it is advantageous to use purified antibody in defined quantity to facilitate assay reproducibility, economy, and reduced interference of nonspecific components as well as improved storage, stability, and bio-conjugation.

Detection of shiga toxins by lateral flow assay.

Shiga toxin-producing Escherichia coli (STEC) produce shiga toxins (Stxs) that can cause human disease and death. The contamination of food products with STEC represents a food safety problem that necessitates rapid and effective detection strategies to mitigate risk. In this manuscript, we report the development of a colorimetric lateral flow assay (LFA) for the rapid detection of Stxs in <10 min using a pair of monoclonal antibodies that bind epitopes common to Stx1 and six Stx2 variants.

Rapid and selective detection of botulinum neurotoxin serotype-A and -B with a single immunochromatographic test strip.

Botulinum neurotoxins (BoNT) are the most potent toxins known. Produced by Clostridium botulinum, BoNTs are classified into seven, antigenically distinct serotypes, designated A-G. The toxin acts to inhibit acetylcholine release, resulting in paralysis and death.

A rapid method to improve protein detection by indirect ELISA.

The enzyme-linked immunosorbant assay (ELISA) is a rapid, high-throughput, quantitative immunoassay for the selective detection of target antigens. The general principle behind an ELISA is antibody mediated capture and detection of an antigen with a measurable substrate. Numerous incarnations of the ELISA have resulted in its commercialization for sensitive diagnostic applications using a variety of detection platforms.

Greenhouse gas, animal performance, and bacterial population structure responses to dietary monensin fed to dairy cows.

The present study investigated the effects of a feed additive and rumen microbial modifier, monensin sodium (monensin), on selected variables in lactating dairy cows. Monensin fed cows (MON, 600 mg d(-1)) were compared with untreated control cows (CON, 0 mg d(-1)) with respect to the effects of monensin on the production of three greenhouse gases (GHG), methane (CH(4)), nitrous oxide (N(2)O), and carbon dioxide (CO(2)), along with animal performance (dry matter intake; DMI), milk production, milk components, plasma urea nitrogen (PUN), milk urea nitrogen (MUN), and the microbial population structure of fresh feces. Measurements of GHG were collected at Days 14 and 60 in an environmental chamber simulating commercial dairy freestall housing conditions.

Effect of dietary monensin on the bacterial population structure of dairy cattle colonic contents.

To determine the effect of monensin, a carboxylic polyether ionophore antibiotic, on the bacterial population structure of dairy cattle colonic contents, we fed six lactating Holstein cows a diet containing monensin (600 mg day(-1)) or an identical diet without monensin. Fresh waste samples were taken directly from the animals once a month for 3 months and assayed for their bacterial population structure via 16S rRNA gene sequence analysis. In total 6,912 16S rRNA genes were examined, comprising 345 and 315 operational taxonomic units (OTUs) from the monensin fed and control animals, respectively.

Identification and functional characterization of the iron-dependent regulator (IdeR) of Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease in cattle and sheep, has unique iron requirements in that it is mycobactin-dependent for cultivation in vitro. The iron-dependent regulator (IdeR) is a well-characterized global regulator responsible for maintaining iron homeostasis in Mycobacterium tuberculosis (MTB). We identified an orthologous segment in the MAP genome, MAP2827, with >93 % amino acid identity to MTB IdeR.