Microbiology Commons^™

Evaluation Of Horizontal Gene Transfer Between Genetically Engineered Cyanobacteria And Gram-Negative Bacteria, Andriana Chrysovalanti Zourou

Chemistry & Biochemistry Theses & Dissertations

As the world population is increasing and societies become more technology driven, there is an imperative to develop ‘green energy’ sources to protect our planet. Cyanobacteria that have been genetically engineered to produce organic compounds that may be burnt as fuels show great potential, as they are an environmentally friendly and self-renewable, net carbon-neutral option. However, there are potential risks in the development and use of genetically modified organisms (GMOs). We need to understand in advance the risks that GMOs may pose to our environment and to animal and human health. This will enable experimental procedures, containment strategies and policies …

Directed Genome Evolution To Identify Genes For Macrophage Survival By Staphylococcus Agnetis, Sonali Lakshika Anne Lenaduwe Lokuge

Graduate Theses and Dissertations

Bacterial Chondronecrosis with Osteomyelitis (BCO) is a debilitating infection that negatively impacts animal welfare and costs the broiler industry billions of dollars annually. We have previously isolated Staphylococcus agnetis 908 from BCO samples obtained from broilers at the University of Arkansas research farm. This isolate can induce BCO lameness at greater than 50% in broilers exposed to the pathogen in drinking water. We found that S. agnetis 908 is capable of surviving and escaping macrophages compared to a closely related cattle isolate,1379. Through Directed Genome Evolution (DGE) we identified that this difference is at least partially associated with an alanine …

Bispecific Anti-Hiv Immunoadhesins That Bind Gp120 And Gp41 Have Broad And Potent Hiv-Neutralizing Activity, Seth H. Pincus, Ryan B. Craig, Lauren Weachter, Celia C. Labranche, Rafiq Nabi, Connie Watt, Mark Raymond, Tami Peters, Kejing Song, Grace A. Maresh, David C. Montefiori, Pamela A. Kozlowski

School of Graduate Studies Faculty Publications

We have constructed bispecific immunoglobulin-like immunoadhesins that bind to both the HIV-envelope glycoproteins: gp120 and gp41. These immunoadhesins have N terminal domains of human CD4 engrafted onto the N-terminus of the heavy chain of human anti-gp41 mAb 7B2. Binding of these constructs to recombinant Env and their antiviral activities were compared to that of the parental mAbs and CD4, as well as to control mAbs. The CD4/7B2 constructs bind to both gp41 and gp140, as well as to native Env expressed on the surface of infected cells. These constructs deliver cytotoxic immunoconjugates to HIV-infected cells, but not as well as …

Development Of Vaccines And Antivirals Against Zika Virus, Aryamav Pattnaik

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The re-emergence of Zika virus (ZIKV), an arbovirus, poses a major global human health concern because of its ability to cause congenital abnormalities and neurological diseases. While many candidate vaccines and antiviral drugs are in the developmental pipeline, none have been approved for use against ZIKV infection. This dissertation describes the characterization of one vaccine and two antiviral drug candidates against ZIKV infection. A bacterial ferritin-based nanoparticle vaccine, termed zDIII-F, is designed to display multiple copies of ZIKV E protein domain III on its surface. These stable nanoparticles are shown to induce robust antibody-mediated protection against lethal ZIKV infection in …

Protective Role Of Palmitoleate Against Zika Virus-Induced Apoptosis In Placental Trophoblasts, Philma Glora Muthuraj

College of Education and Human Sciences: Dissertations, Theses, and Student Research

Zika virus (ZIKV) infection in pregnant women causes congenital Zika syndrome which involves birth defects such as intrauterine growth restriction (IUGR), retinal defects and microcephaly in the fetus. ZIKV infection results in placental pathology which plays a crucial role in disease transmission from mother to fetus. Currently, there is no Food and Drug Administration (FDA) approved vaccine or therapeutic drug against ZIKV. In this thesis, we have elucidated the cell signaling pathways connected to ZIKV-induced apoptosis in trophoblasts and also the protective effect of palmitoleate against ZIKV-induced apoptosis. First, we demonstrated the molecular mechanism behind ZIKV-induced apoptosis using an in …

A Yeast Expressed Rbd-Based Sars-Cov-2 Vaccine Formulated With 3m-052-Alum Adjuvant Promotes Protective Efficacy In Non-Human Primates, Maria Pino, Talha Abid, Susan Pereira Ribeiro, Venkata Viswanadh Edara, Katharine Floyd, Justin C. Smith, Muhammad Bilal Latif, Gabriela Pacheco-Sanchez, Debashis Dutta, Shelly Wang, Sanjeev Gumber, Shannon Kirejczyk, Joyce Cohen, Rachelle L. Stammen, Sherrie M. Jean, Jennifer S. Wood, Fawn Connor-Stroud, Jeroen Pollet, Wen Hsiang Chen, Junfei Wei, Bin Zhan, Jungsoon Lee, Zhuyun Liu, Ulrich Strych, Neeta Shenvi, Kirk Easley, Daniela Weiskopf, Alesandro Sette

School of Graduate Studies Faculty Publications

Ongoing SARS-CoV-2 vaccine development is focused on identifying stable, cost-effective, and accessible candidates for global use, specifically in low and middle-income countries. Here, we report the efficacy of a rapidly scalable, novel yeast expressed SARS-CoV-2 specific receptor-binding domain (RBD) based vaccine in rhesus macaques. We formulated the RBD immunogen in alum, a licensed and an emerging alum adsorbed TLR-7/8 targeted, 3M-052-alum adjuvants. The RBD+3M-052-alum adjuvanted vaccine promoted better RBD binding and effector antibodies, higher CoV-2 neutralizing antibodies, improved Th1 biased CD4+T cell reactions, and increased CD8+ T cell responses when compared to the alum-alone adjuvanted vaccine. RBD+3M-052-alum induced a significant …

The Persistence Of Human Norovirus Surrogate In Leafy Greens Production System, Wenjun Deng

Graduate Theses and Dissertations

Since mostly consumed raw, foodborne pathogen contamination of leafy greens has led to a large number of foodborne disease outbreaks and illnesses each year in the United States. Human noroviruses (hNoV) are the most common viral pathogen transmitted by leafy greens. In this dissertation, the persistence of the hNoV surrogate Tulane virus (TV) on pre-harvest lettuce and microgreens was investigated. Lettuces are the most studied leafy green model, while previous hNoV research has mainly focused on the post-harvest stage of production. Here, pre-harvest hydroponically grown lettuce were used to determine TV persistence on leafy greens. After inoculation on leaves at …

Functional And Phenotypic Evaluation Of Murine Cytomegalovirus Infection Of The Heart, Cassandra Marie Bonavita

LSU Doctoral Dissertations

Human Cytomegalovirus (HCMV) is a ubiquitous pathogen which infects 50-90% of the world population and results in lifelong latent infection with periodic transient reactivations. In immunocompetent populations acute infection is generally asymptomatic but seropositivity is associated with many chronic diseases, including cardiovascular disease. Despite 139 years of research into HCMV pathogenesis there is still no licensed vaccine and many gaps in the general knowledge of the virus. This includes the role of long term infection as it pertains to chronic diseases. In these studies, murine cytomegalovirus (MCMV) was used as a model of HCMV pathogenesis in the heart. MCMV shares …

Lead And Cadmium Detoxification By Halophilic Bacteria Isolated From Solar Salterns In Lebanon, Nadine G. Zmorrod N. G. Zmorrod, Nawal Al Hakawati, Ranya A. Amer, Hoda H. Yusef

BAU Journal - Science and Technology

Water contamination by heavy metals has gained considerable attention globally. These inorganic pollutants can enter the aquatic environments via different routes thereby threatening biodiversity and human health. Lead and cadmium are hazardous pollutants where their removal by traditional techniques is identified to be costly and ineffective. However, bioremediation by extremophilic microorganisms is considered to be a promising technique as they have considerable potentials to grow in harsh environmental conditions. The present study deals with the isolation of halophilic bacterial isolates from solar salterns in North Lebanon. The isolate H1S9 was identified as Halomonas venusta H9 after showing tolerance to 500 …

Transcriptomic And Functional Investigation Of Bacterial Biofilm Formation, Brooke R. Nemec

USF Tampa Graduate Theses and Dissertations

Staphylococcus aureus and Acinetobacter baumannii are two highly successful human pathogens, which have adopted very different, but effective survival strategies. The success of S. aureus is attributed to the tight regulation of an arsenal of virulence factors. Conversely, A. baumannii lacks what would be considered traditional virulence factors and, instead, has developed a high tolerance for environmental stress, which allows it to persist in unforgiving environments, including nosocomial settings and the human body. One common characteristic of these two organisms is their proclivity for biofilm formation. Herein, we discuss the diverse mechanisms governing biofilm formation for A. baumannii and S. …

Telomeric And Sub-Telomeric Structure And Implications In Fungal Opportunistic Pathogens, Raffaella Diotti, Michelle Esposito, Chang Hui Shen

Publications and Research

Telomeres are long non-coding regions found at the ends of eukaryotic linear chromosomes. Although they have traditionally been associated with the protection of linear DNA ends to avoid gene losses during each round of DNA replication, recent studies have demonstrated that the role of these sequences and their adjacent regions go beyond just protecting chromosomal ends. Regions nearby to telomeric sequences have now been identified as having increased variability in the form of duplications and rearrangements that result in new functional abilities and biodiversity. Furthermore, unique fungal telomeric and chromatin structures have now extended clinical capabilities and understanding of pathogenicity …

Exploring Prokaryotic And Viral Communities Within Florida’S Freshwater Springs, Kema Saydoon Malki

USF Tampa Graduate Theses and Dissertations

Florida’s freshwater springs serve as a crucial ecosystem for the state, having both ecological and economic importance. Springs occur where there is an opening where aquifers discharge groundwater. Florida’s springs are fed by the Floridan aquifer system (FAS), which underlies the entire state and extends into several neighboring states. The FAS provides potable water to over 10 million people. The constant flow of groundwater through the springs feeds major river systems and creates unique freshwater systems that support numerous plants and animals. The combination of natural beauty and charismatic fauna attract tourism and generate millions of dollars in revenue for …

A Bacterial Inflammation Sensor Regulates C-Di-Gmp Signaling, Adhesion, And Biofilm Formation, Arden Perkins, Dan A. Tudorica, Raphael D. Teixeira, Tilman Schirmer, Lindsay Zumwalt, O. Maduka Ogba, C. Keith Cassidy, Phillip J. Stansfeld, Karen Guillemin

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Bacteria that colonize animals must overcome, or coexist, with the reactive oxygen species products of inflammation, a front-line defense of innate immunity. Among these is the neutrophilic oxidant bleach, hypochlorous acid (HOCl), a potent antimicrobial that plays a primary role in killing bacteria through nonspecific oxidation of proteins, lipids, and DNA. Here, we report that in response to increasing HOCl levels, Escherichia coli regulates biofilm production via activation of the diguanylate cyclase DgcZ. We identify the mechanism of DgcZ sensing of HOCl to be direct oxidation of its regulatory chemoreceptor zinc-binding (CZB) domain. Dissection of CZB signal transduction reveals that …

Host Phylogeny And Diet Shape Gut Microbial Communities Within Bamboo-Feeding Insects, Kuanguan Huang, Jie Wang, Junhao Huang, Shouke Zhang, Alfried P. Vogler, Quanquan Liu, Yongchun Li, Maowei Yang, You Li, Xuguo Zhou

Entomology Faculty Publications

The gut microbiome plays an important role in a host’s development and adaption to its dietary niche. In this study, a group of bamboo-feeding insects are used to explore the potential role of the gut microbiota in the convergent adaptation to extreme diet specialization. Specifically, using a 16S rRNA marker and an Illumina sequencing platform, we profiled the microbial communities of 76 gut samples collected from nine bamboo-feeding insects, including both hemimetabolous (Orthoptera and Hemiptera) and holometabolous (Coleoptera and Lepidoptera) species, which are specialized in three distinct dietary niches: bamboo leaf, shoot, and sap. The gut microbiota of these insects …

Oral Vaccination Approaches For Anti-Shiv Immunity, Erandi Velarde De La Cruz, Lingyun Wang, Deepanwita Bose, Sailaja Gangadhara, Robert L. Wilson, Rama R. Amara, Pamela A. Kozlowski, Anna Aldovini

School of Graduate Studies Faculty Publications

We modified a Sabin Oral Poliovirus Vaccine (OPV) vector to permit secretion of the antigens of interest with the goal of improving anti-HIV Env humoral responses in a SHIV mucosal immunization composed of DNA and recombinant OPVs. We evaluated stimulation of systemic and mucosal cell-mediated and humoral immunity in Rhesus macaques by two regimens, both involving a prime with a SHIVBG505 DNA construct producing non-infectious particles formulated in lipid nanoparticles, administered in the oral cavity, and two different viral vector boostings, administered in the oral cavity and intestinally. Group 1 was boosted with rMVA-SHIVBG505, expressing SIV Gag/Pol and HIVBG505 Env. …

The Seasonal Flux And Fate Of Dissolved Organic Carbon Through Bacterioplankton In The Western North Atlantic, Nicholas Baetge, Michael J. Behrenfeld, James Fox, Kimberly H. Halsey, Kristina D.A. Mojica, Anai Novoa, Brandon M. Stephens, Craig A. Carlson

Faculty Publications

The oceans teem with heterotrophic bacterioplankton that play an appreciable role in the uptake of dissolved organic carbon (DOC) derived from phytoplankton net primary production (NPP). As such, bacterioplankton carbon demand (BCD), or gross heterotrophic production, represents a major carbon pathway that influences the seasonal accumulation of DOC in the surface ocean and, subsequently, the potential vertical or horizontal export of seasonally accumulated DOC. Here, we examine the contributions of bacterioplankton and DOM to ecological and biogeochemical carbon flow pathways, including those of the microbial loop and the biological carbon pump, in the Western North Atlantic Ocean (∼39–54°N along ∼40°W) …

Candida Cell‐Surface‐Specific Monoclonal Antibodies Protect Mice Against Candida Auris Invasive Infection, Jonothan Rosario‐Colon, Karen Eberle, Abby Adams, Evan Courville, Hong Xin

School of Graduate Studies Faculty Publications

Candida auris is a multidrug‐resistant fungal pathogen that can cause disseminated bloodstream infections with up to 60% mortality in susceptible populations. Of the three major classes of antifungal drugs, most C. auris isolates show high resistance to azoles and polyenes, with some clinical isolates showing resistance to all three drug classes. We reported in this study a novel approach to treating C. auris disseminated infections through passive transfer of monoclonal antibodies (mAbs) targeting cell surface antigens with high homology in medically important Candida spe-cies. Using an established A/J mouse model of disseminated infection that mimics human candidi-asis, we showed that …

The Rice Blast Fungus Morgs1 Functioning In Camp Signaling And Pathogenicity Is Regulated By Casein Kinase Mock2 Phosphorylation And Modulated By Membrane Protein Moemc2, Rui Yu, Xuetong Shen, Muxing Liu, Xinyu Liu, Ziyi Yin, Xiao Li, Wanzhen Feng, Jiexiong Hu, Haifeng Zhang, Xiaobo Zheng, Ping Wang, Zhengguang Zhang

School of Graduate Studies Faculty Publications

GTP-binding protein (G-protein) and regulator of G-protein signaling (RGS) mediated signal transduction are critical in the growth and virulence of the rice blast pathogen Magnaporthe oryzae. We have previously reported that there are eight RGS and RGS-like proteins named MoRgs1 to MoRgs8 playing distinct and shared regulatory functions in M. oryzae and that MoRgs1 has a more prominent role compared to others in the fungus. To further explore the unique regulatory mechanism of MoRgs1, we screened a M. oryzae cDNA library for genes encoding MoRgs1-interacting proteins and identified MoCkb2, one of the two regulatory subunits of the casein kinase (CK) …

Sars-Cov-2 Vaccines Elicit Durable Immune Responses In Infant Rhesus Macaques, Carolina Garrido, Alan D. Curtis, Maria Dennis, Sachi H. Pathak, Hongmei Gao, David Montefiori, Mark Tomai, Christopher B. Fox, Pamela A. Kozlowski, Trevor Scobey, Jennifer E. Munt, Michael L. Mallory, Pooja T. Saha, Michael G. Hudgens, Lisa C. Lindesmith, Ralph S. Baric, Olubukola M. Abiona, Barney S. Graham, Kizzmekia S. Corbett, Darin Edwards

School of Graduate Studies Faculty Publications

The inclusion of infants in the SARS-CoV-2 vaccine rollout is important to prevent severe complications of pediatric SARS-CoV-2 infections and to limit transmission and could possibly be implemented via the global pediatric vaccine schedule. However, age-dependent differences in immune function require careful evaluation of novel vaccines in the pediatric population. Toward this goal, we assessed the safety and immunogenicity of two SARSCoV- 2 vaccines. Two groups of eight infant rhesus macaques (RMs) were immunized intramuscularly at weeks 0 and 4 with stabilized prefusion SARS-CoV-2 S-2P spike (S) protein encoded by mRNA encapsulated in lipid nanoparticles (mRNA-LNP) or the purified S …

Deacylated Trna Accumulation Is A Trigger For Bacterial Antibiotic Persistence Independent Of The Stringent Response, Whitney N. Wood, Kyle Mohler, Jesse Rinehart, Michael Ibba

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Bacterial antibiotic persistence occurs when bacteria are treated with an antibiotic and the majority of the population rapidly dies off, but a small subpopulation enters into a dormant, persistent state and evades death. Diverse pathways leading to nucleoside triphosphate (NTP) depletion and restricted translation have been implicated in persistence, suggesting alternative redundant routes may exist to initiate persister formation. To investigate the molecular mechanism of one such pathway, functional variants of an essential component of translation (phenylalanyltRNA synthetase [PheRS]) were used to study the effects of quality control on antibiotic persistence. Upon amino acid limitation, elevated PheRS quality control led …

Prophylactic Phage Therapy In Infant Rabbits Model Of Cholera, Sadeq Ag Kaabi Asst Prof Dr

Karbala International Journal of Modern Science

A number of 8 phages lytic of V. cholerae O1- biotype-EI-Tor, serotype Inaba were selected for the formulation of a phage cocktail. A phage cocktail composed of 8 phages was prepared and showed 100% inhibition of V. cholerae isolates comparing with a percentage ranged from 0-75% for every single phage in growth inhibition assay. The potential activity of phage cocktail of prophylactic therapy for infant rabbits model of cholera was evaluated through phage retention time and length of phage prophylaxis studies. Results have been showed that phage cocktail was potent in the prevention of development of cholera in infant rabbits …

Non-B Dna-Forming Motifs Promote Mfd-Dependent Stationary-Phase Mutagenesis In Bacillus Subtilis, Tatiana Ermi, Carmen Vallin, Ana Gabriela Regalado García, Moises Bravo, Ismaray Fernandez Cordero, Holly Anne Martin, Mario Pedraza-Reyes, Eduardo Robleto

Life Sciences Faculty Research

Transcription-induced mutagenic mechanisms limit genetic changes to times when expression happens and to coding DNA. It has been hypothesized that intrinsic sequences that have the potential to form alternate DNA structures, such as non-B DNA structures, influence these mechanisms. Non-B DNA structures are promoted by transcription and induce genome instability in eukaryotic cells, but their impact in bacterial genomes is less known. Here, we investigated if G4 DNA-and hairpin-forming motifs influence stationary-phase mutagenesis in Bacillus subtilis. We developed a system to measure the influence of non-B DNA on B. subtilis stationary-phase mutagenesis by deleting the wild-type argF at its chromosomal …

Microbial Screening Reveals Oral Site-Specific Locations Of The Periodontal Pathogen Selenomonas Noxia, Jaydene Mcdaniel, Steven Mcdaniel, Beanca Jhanine Samiano, Matthew Marrujo, Karl Kingsley, Katherine M. Howard

Dental Medicine Faculty Research

Introduction: Selenomonas noxia (SN) is an important periodontal pathogen, associated with gingivitis and periodontitis. Many studies have found associations between SN and indicators of poor health outcomes, such as smoking, low socioeconomic status and obesity. However, less is known about the prevalence of this organism and more specifically about other oral site-specific locations that may harbor this organism. Methods: Using an existing patient repository (n = 47) of DNA isolated from saliva and other oral sites (n = 235), including the dorsum of the tongue, lower lingual incisor, upper buccal molar and gingival crevicular fluid (GCF), molecular screening for SN …

Prevalence And Risk Factors Associated With Multi-Drug Resistant Organisms (Mdro) Carriage Among Pediatric Patients At The Time Of Admission In A Tertiary Care Hospital Of A Developing Country. A Cross-Sectional Study, Sonia Qureshi, Noshi Maria, Mohammad Zeeshan, Seema Irfan, Farah Naz Qamar

Department of Paediatrics and Child Health

Background: The rise of Multidrug-resistant organisms (MDROs) poses a considerable burden on the healthcare systems, particularly in low-middle income countries like Pakistan. There is a scarcity of data on the carriage of MDRO particularly in the pediatrics population therefore, we aimed to determine MDRO carriage in pediatric patients at the time of admission to a tertiary care hospital in Karachi, Pakistan, and to identify the risk factors associated with it.
Methods: A cross-sectional study conducted at the pediatric department of Aga Khan University Hospital (AKUH) from May to September 2019 on 347 children aged 1-18 years. For identification of MDRO …

Development And Validation Of An Isothermal Amplification Assay For Eastern Equine Encephalitis Virus, Mikayla D. Maddison

USF Tampa Graduate Theses and Dissertations

Eastern Equine Encephalitis virus (EEEV) is a neurotrophic alphavirus for which there is no effective treatment or vaccine for humans. Periodic outbreaks in the Eastern United States represent an ongoing public health problem; Florida serves as the reservoir for EEEV for the rest of the country. Reverse transcription-polymerase chain reaction (RT-PCR) is the current gold standard for molecular diagnostic testing of the presence of EEEV in vectors. However, RT-PCR is technically complex and can be difficult for mosquito control districts to utilize. In order to provide a simple, cost-effective alternative for mosquito surveillance and control, a novel one step reverse …

Draft Genome Sequences Of 13 Vibrio Cholerae Strains From The Rio Grande Delta, Jeffrey W. Turner, Jorge Duran-Gonzalez, David A. Laughlin, Daniel Unterweger, David Silva, Boris Ermolinsky, Stefan Pukatzki, Daniele Provenzano

School of Integrative Biological & Chemical Sciences (Formerly Dept. of Biology)

Vibrio cholerae is the etiologic agent of cholera, an acute and often fatal diarrheal disease that affects millions globally. We report the draft genome sequences of 13 non-O1/O139 V. cholerae strains isolated from the Rio Grande Delta in Texas. These genomes will aid future analyses of environmental serovars.

Draft Genome Sequences Of 13 Vibrio Cholerae Strains From The Rio Grande Delta, Jeffrey W. Turner, Jorge Duran-Gonzalez, David A. Laughlin, Daniel Unterweger, David Silva, Boris Ermolinsky, Stefan Pukatzki, Daniele Provenzano

Publications and Research

Vibrio cholerae is the etiologic agent of cholera, an acute and often fatal diarrheal disease that affects millions globally. We report the draft genome sequences of 13 non-O1/O139 V. cholerae strains isolated from the Rio Grande Delta in Texas. These genomes will aid future analyses of environmental serovars.

Global Analysis Of High Cited Papers On “Impact Of Covid-19 On Mental Health” During 2020-21, Ghouse Modin Nabeesab Mamdapur Mr., Brij Mohan Gupta Dr., Sandeep Grover Dr.

Library Philosophy and Practice (e-journal)

A bibliometric analysis based on 160 highly cited papers extracted from the Scopus international database was carried out to provide insights into literature characteristics and publication performances of various participating actors on “Impact of COVID-19 on Mental Health”. Quantitative and qualitative Indicators were applied to measure the productivity and citation impact of most productive participating countries, organizations, authors, journals and significant keywords and to visualise and measure collaborative interaction among them using VOSviewer software. Results obtained from this study can provide valuable information for researchers and policy-makers to identify present and future hotspots in research on “Impact of COVID-19 on …

Development Of A Novel, High Resolution Melting Analysis Based Genotyping Method For Cryptosporidium Parvum, Jennifer K. O'Leary, Liam Blake, Gerard D. Corcoran, Roy D. Sleator, Brigid Lucey

Department of Biological Sciences Publications

This study employed the post-real-time PCR application, high resolution melting (HRM) analysis, in order to differentiate between characterised clinical and reference Cryptosporidium parvum samples obtained from Cork University Hospital (Cork, Ireland) and the Cryptosporidium Reference Unit (Swansea, Wales). A sample set composed of 18 distinct C. parvum gp60-subtypes of the IIa gp60-subtype family (an allele family accounting for over 80% of all cryptosporidiosis cases in Ireland) was employed. HRM analysis-based interrogation of the gp60, MM5 and MS9-Mallon tandem repeat loci was found to completely differentiate between 10 of the 18 studied gp60-subtypes. The remaining eight gp60-subtypes were differentiated into three …

A Novel Genotyping Method For Cryptosporidium Hominis, Jennifer K. O'Leary, Liam Blake, Gerard D. Corcoran, Roy D. Sleator, Brigid Lucey

Department of Biological Sciences Publications

Cryptosporidiosis remains the leading protozoan induced cause of diarrhoea-associated mortality worldwide. Cryptosporidium hominis, the anthroponotically transmitted species within the Cryptosporidium genus, contributes significantly to the global burden of infection, accounting for the majority of clinical cases in many countries. This study applied high resolution melting analysis, a post-real-time PCR application, to the differentiation of six globally prevalent C. hominis gp60-subtypes. This novel method targeted three microsatellite, tandem repeat containing genetic markers, gp60, the genetic marker upon which current Cryptosporidium subtype nomenclature is based, MSB, and MSE, by which to differentiate between C. hominis isolates. This multi-locus approach successfully differentiated between …