2019-2020 Dean\u27s Showcase No. 3

https://spiral.lynn.edu/conservatory_deansshowcase/1073/thumbnail.jp

Elution independent collection device (EICD) for rapid collection of Anaplasma marginale DNA from blood samples

Currently utilized molecular detection methods are based mainly on nucleic acid extraction, amplification, and detection procedures that may require costly equipment, numerous reagents, and highly trained personnel. These requirements make diagnostic tests expensive, time-consuming, and not suitable for point-of-care applications. There is an increasing demand for simple, low-cost portable technologies. To overcome these challenges, a paper-based elution independent collection device (EICD) was designed to collect microorganisms and recover nucleic acids for molecular biology applications with minimal steps. In this study, we demonstrate a simpler Anaplasma marginale detection that uses an EICD for nucleic acid collection combined with recombinase polymerase amplification (RPA), and a lateral flow dipstick for detection of the specified target. A pre-lysis blood treatment was optimized that uses Triton X-100 lysis buffer and bovine serum album in wash buffer. Blood samples were incubated for 5 min at room temperature and run through the EICD. Four 1-mm diameter discs excised from EICD were used as template in basic RPA and lateral flow (nfo) (endonuclease IV) RPA assays. Each disc of soluble central membrane (SCM) carried circa 0.249 pg/ul of Anaplasma DNA. The percentage of nucleic acid recoverable from the SCM ranged between 60% - 70%. Blood samples infected with A. marginale were treated with Triton X-100 pre-lysis protocol. All samples tested positive by PCR and RPA methods. EICD-driven collection of blood samples is a practical method successfully adapted to detect Anaplasma spp. or blood-borne pathogen DNA and has potential for point-of-care detection in resource-limited settings.Peer reviewedEntomology and Plant PathologyBiosecurity and Microbial Forensic

Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health

Anaplasma marginale, A. ovis, and A. phagocytophilum are the causative agents of bovine anaplasmosis, ovine anaplasmosis, and granulocytic anaplasmosis, respectively. The gold standard for diagnosis of post-acute and long-term persistent infections is the serological cELISA, which does not discriminate between Anaplasma species and requires highly equipped laboratories and trained personnel. This study addresses the development of a rapid, isothermal, sensitive, species-specific RPA assays to detect three Anaplasma species in blood and cELISA A. marginale-positive serum samples. Three RPA primer and probe sets were designed targeting msp4 genes of each Anaplasma species and the internal control (GAPDH gene) for each assay. The limit of detection of gel-based or RPA-basic assays is 8.99 x 104 copies/ul = A. marginale, 5.04 x� 106 copies/ul = A. ovis, and 4.58 x� 103 copies/ul = A. phagocytophilum, and for each multiplex lateral flow or RPA-nfo assays is 8.99 x� 103 copies/ul of A. marginale, 5.04 x� 103 copies/ul of A. ovis, 4.58 x� 103 copies/ul of A. phagocytophilum, and 5.51 x� 103 copies/ul of internal control (GAPDH). Although none of the 80 blood samples collected from Oklahoma cattle were positive, the RPA-nfo assays detected all A. marginale cattle blood samples with varying prevalence rates of infection, 83% of the 24 cELISA A. marginale-positive serum samples, and all A. phagocytophilum cell culture samples. Overall, although early detection of three Anaplasma species was not specifically addressed, the described RPA technique represents an improvement for detection of three Anaplasma in regions where access to laboratory equipment is limited.Peer reviewedEntomology and Plant PathologyBiosecurity and Microbial Forensic

Bycatch species composition over time by tuna purse-seine fishery in the eastern tropical Atlantic Ocean.

Post-print

Convergencias transculturales en el Caribe y Centroamérica

Editores: Mauricio Chaves, Werner Mackenbach, Héctor Pérez BrignoliLos ensayos recopilados en este libro se basan en trabajos presentados en el Simposio Internacional: Convergencias transculturales en el Caribe. Literatura, arte, cultura, historia, comunicación realizada en noviembre de 2015 en la Universidad de Costa Rica.4 Exploran algunas dimensiones de los “entrecruzamientos”, “branchementes” y “entanglements” entre Centroamérica y el Caribe).UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Sociales::Centro de Investigaciones Históricas de América Central (CIHAC