Microbiology Commons^™

Microbial Nitrogen Limitation Increases Decomposition, Joseph M. Craine, Carl Morrow, Noah Fierer

Dartmouth Scholarship

With anthropogenic nutrient inputs to ecosystems increasing globally, there are long-standing, fundamental questions about the role of nutrients in the decomposition of organic matter. We tested the effects of exogenous nitrogen and phosphorus inputs on litter decomposition across a broad suite of litter and soil types. In one experiment, C mineralization was compared across a wide array of plants individually added to a single soil, while in the second, C mineralization from a single substrate was compared across 50 soils. Counter to basic stoichiometric decomposition theory, low N availability can increase litter decomposition as microbes use labile substrates to acquire …

Dissecting Biological “Dark Matter” With Single-Cell Genetic Analysis Of Rare And Uncultivated Tm7 Microbes From The Human Mouth, Yann Marcy, Cleber C. Ouverney, Elisabeth M. Bik, Tina Lösekann, Natalia Ivanova, Hector Garcia Martin, Ernest Szeto, Darren Platt, Philip Hugenholtz, David A. Relman, Stephen R. Quake

Faculty Publications, Biological Sciences

We have developed a microfluidic device that allows the isolation and genome amplification of individual microbial cells, thereby enabling organism-level genomic analysis of complex microbial ecosystems without the need for culture. This device was used to perform a directed survey of the human subgingival crevice and to isolate bacteria having rod-like morphology. Several isolated microbes had a 16S rRNA sequence that placed them in candidate phylum TM7, which has no cultivated or sequenced members. Genome amplification from individual TM7 cells allowed us to sequence and assemble >1,000 genes, providing insight into the physiology of members of this phylum. This approach …

The Distribution Of Sexually-Transmitted Human Papillomaviruses In Hiv Positive And Negative Patients In Zambia, Africa, Christopher Ngandwe, John Lowe, Paula J. Richards, Lara Hause, Charles Wood, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Background: Human Papillomaviruses (HPV) are double-stranded DNA viruses, considered to be the primary etiological agents in cervical intraepithelial neoplasias and cancers. Approximately 15–20 of the 40 mucosal HPVs confer a high-risk of progression of lesions to invasive cancer. In this study, we investigated the prevalence of sexually transmitted HPVs in Human Immunodeficiency Virus (HIV) positive and negative patients in Zambia, Africa. The rate of high-risk HPV genotypes worldwide varies within each country. Thus, we sought to investigate the rates of HPV infection in sub-Saharan Africa and the potential role of HIV in affecting the HPV genotype distribution.

Methods: This retrospective …

Characterization Of A Novel Nuclear Variant Of Bmp2 And Coordinate Regulation Of Col11a2 And Col27a1 By The Transcription Factor Lc-Maf, Jaime Lynn Mayo

Theses and Dissertations

ABSTRACT I CHARACTERIZATION OF A NOVEL NUCLEAR VARIANT OF BMP2

Bone morphogenetic protein 2 (Bmp2) is a signaling protein that was first detected by its ability to induce cartilage and bone formation. It has since been implicated in broad variety of developmental, patterning, and disease processes. To date, Bmp2 has only been known to function as an extracellular signaling molecule. However, we have obtained clear evidence for a nuclear form of Bmp2. This nuclear variant, nBmp2, contains a bipartite NLS that overlaps the site of proteolytic cleavage. The NLS remains intact and functional when translation of Bmp2 initiates from a …

Mammalian Cell Cytotoxicity Analysis Of Soybean Rust Fungicides, Steven L. Daniel, G. L. Hartman, E. D. Wagner, M. J. Plewa

Faculty Research & Creative Activity

The identification of soybean rust, caused by Phakopsora

pachyrhizi H. Sydow & Sydow, in the southern United

States in November 2004 (Schneider et al. 2005 ), in the

Midwest in 2006 (Hartman et al. 2007 ), and elsewhere has

increased the concerns of the impact of the pathogen on US

soybean production (Miles et al. 2007 ). The rapid spread of

P. pachyrhizi and its potential to cause severe yield losses

makes this among the most destructive foliar diseases of

soybean. Yield losses of 20%–60% were reported in Asia,

with losses of 80% reported from experimental plots in

Taiwan (Hartman …

Mammalian Cell Cytotoxicity Analysis Of Soybean Rust Fungicides, Steven Daniel, G. Hartman, E. Wagner, M. Plewa

Faculty Research & Creative Activity

No abstract provided.

Mammalian Cell Cytotoxicity Analysis Of Soybean Rust Fungicides, Steven L. Daniel, G. L. Hartman, E. D. Wagner, M. J. Plewa

Faculty Research & Creative Activity

No abstract provided.

Comparative Functional Analysis Of Aquaporins/Glyceroporins In Mammals And Anurans, Carissa M. Krane, David L. Goldstein

Biology Faculty Publications

Maintenance of fluid homeostasis is critical to establishing and maintaining normal physiology. The landmark discovery of membrane water channels (aquaporins; AQPs) ushered in a new area in osmoregulatory biology that has drawn from and contributed to diverse branches of biology, from molecular biology and genomics to systems biology and evolution, and from microbial and plant biology to animal and translational physiology. As a result, the study of AQPs provides a unique and integrated backdrop for exploring the relationships between genes and genome systems, the regulation of gene expression, and the physiologic consequences of genetic variation. The wide species distribution of …

Chloroviruses Encode A Bifunctional Dcmp-Dctp Deaminase That Produces Two Key Intermediates In Dttp Formation, Yuanzheng Zhang, Frank Maley, Gladys F. Maley, Garry Duncan, David Dunigan, James L. Van Etten

Nebraska Center for Virology: Faculty Publications

The chlorovirus PBCV-1, like many large double-stranded DNA-containing viruses, contains several genes that encode putative proteins involved in nucleotide biosynthesis. This report describes the characterization of the PBCV-1 dCMP deaminase, which produces dUMP, a key intermediate in the synthesis of dTTP. As predicted, the recombinant protein has dCMP deaminase activity that is activated by dCTP and inhibited by dTTP. Unexpectedly, however, the viral enzyme also has dCTP deaminase activity, producing dUTP. Typically, these two reactions are catalyzed by proteins in separate enzyme classes; to our knowledge, this is the first example of a protein having both deaminase activities. Kinetic experiments …

Clear Cell Carcinoma Of Ovary With Associated Mucinous Cystadenoma And Endometriosis, Zeeshan Uddin, Nausheen Yaqoob, Naila Kayani, Sheema H. Hasan

Department of Pathology and Laboratory Medicine

A 45 year old woman presented with right sided ovarian mass with multiple omental deposits and liver metastases. The right ovary was enlarged and showed a partly cystic partly solid cut surface. Histological picture showed clear cell carcinoma with areas of mucinous cystadenoma and endometriosis. Clear cell carcinoma is known to be associated with endometriosis. To the best of author's knowledge, it's association with mucinous cystadenoma has been described only once in the literature, where clear cell carcinoma was shown to be associated with mucinous cystadenoma without any evidence of endometriosis.

Serotype Association And Regulation Of The Hyaluronate Lyase Gene Of Streptococcus Pyogenes, Martha Stokes

Theses and Dissertations in Biomedical Sciences

S. pyogenes expresses many virulence factors, controlled by a complex regulatory network. These include hyaluronate lyase, an enzyme that degrades hyaluronic acid, a major component of the human extracellular matrix. Paradoxically, hyaluronic acid is also the sole component of the bacterium's capsule, a primary defense against the host immune response. The hylA gene, which encodes the enzyme, has been shown to take three structural forms: a full-length gene, one containing a 3' deletion and a gene that produces a prematurely truncated protein. This work was intended to show that the structure of hylA is associated with serotype, and …

Bifa, A Cyclic-Di-Gmp Phosphodiesterase, Inversely Regulates Biofilm Formation And Swarming Motility By Pseudomonas Aeruginosa Pa14, Sherry L. Kuchma, Kimberly M. Brothers, Judith H. Merritt, Nicole T. Liberati, Frederick M. Ausubel, George A. O'Toole

Dartmouth Scholarship

The intracellular signaling molecule, cyclic-di-GMP (c-di-GMP), has been shown to influence bacterial behaviors, including motility and biofilm formation. We report the identification and characterization of PA4367, a gene involved in regulating surface-associated behaviors in Pseudomonas aeruginosa. The PA4367 gene encodes a protein with an EAL domain, associated with c-di-GMP phosphodiesterase activity, as well as a GGDEF domain, which is associated with a c-di-GMP-synthesizing diguanylate cyclase activity. Deletion of the PA4367 gene results in a severe defect in swarming motility and a hyperbiofilm phenotype; thus, we designate this gene bifA, for biofilm formation. We show that BifA localizes to the inner …

Detection Of Panulirus Argus Virus 1 In Caribbean Spiny Lobsters, Megan M. Montgomery-Fullerton, Roland Cooper, Kathryn M. Kauffman, Jeffrey D. Shields, Robert E. Ratzlaff

Collected Faculty Scholarship

Panulirus argus Virus 1 (PaV1) is a pathogenic virus that infects Caribbean spiny lobsters P. argus in the Florida Keys. We have developed a PCR detection assay for PaV1 for the purpose of studying the natural history of the virus and for monitoring the prevalence of infection. The detection of the virus in hemolymph and other tissues is based on the PCR amplification of a 499 bp product using specific primers designed from a cloned fragment of the PaV1 genome. The sensitivity limit for the assay was 1.2 fg of purified viral DNA. The PaV1 primers did not react with …

Epstein-Barr Virus Inhibits Kaposi’S Sarcoma-Associated Herpesvirus Lytic Replication In Primary Effusion Lymphomas, Dongsheng Xu, Jun Zhang, Tricia Coleman, Ashley Fagot, Catherine Kotalik, Lingjun Zhao, Pankaj Trivedi, Clinton J. Jones, Luwen Zhang

Nebraska Center for Virology: Faculty Publications

The majority of AIDS-associated primary effusion lymphomas (PEL) are latently infected with both Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV). PELs harboring two viruses have higher oncogenic potential, suggesting functional interactions between EBV and KSHV. The KSHV replication and transcription activator (K-RTA) is necessary and sufficient for induction of KSHV lytic replication. EBV latent membrane protein 1 (LMP-1) is essential for EBV transformation and establishment of latency in vitro. We show EBV inhibits chemically induced KSHV lytic replication, in part because of a regulatory loop in which K-RTA induces EBV LMP-1 and LMP-1 in turn inhibits K-RTA expression and …

The Transcriptional Repressor K-Rbp Modulates Rta-Mediated Transactivation And Lytic Replication Of Kaposi’S Sarcoma-Associated Herpesvirus, Zhilong Yang, Charles Wood

Nebraska Center for Virology: Faculty Publications

The replication and transcription activator (RTA) protein of Kaposi’s sarcoma (KS)-associated herpesvirus (KSHV)/human herpesvirus 8 functions as the key regulator to induce KSHV lytic replication from latency through activation of the lytic cascade of KSHV. Elucidation of the host factors involved in RTA-mediated transcriptional activation is pivotal for understanding the transition between viral latency and lytic replication. KSHV-RTA binding protein (K-RBP) was previously isolated as a cellular RTA binding protein of unknown function. Sequence analysis showed that K-RBP contains a Kruppel-associated box (KRAB) at the N terminus and 12 adjacent zinc finger motifs. In similarity to other KRAB-containing zinc finger …

Apres Le Deluge: Microbial Landscape Of New Orleans After The Hurricanes, Fred C. Dobbs

OES Faculty Publications

No abstract provided.

Parasites Alter Community Structure, Chelsea L. Wood, James E. Byers, Kathryn L. Cottingham, Irit Altman

Dartmouth Scholarship

Parasites often play an important role in modifying the physiology and behavior of their hosts and may, consequently, mediate the influence hosts have on other components of an ecological community. Along the northern Atlantic coast of North America, the dominant herbivorous snail Littorina littorea structures rocky intertidal communities through strong grazing pressure and is frequently parasitized by the digenean trematode Cryptocotyle lingua. We hypothesized that the effects of parasitism on host physiology would induce behavioral changes in L. littorea, which in turn would modulate L. littorea's influence on intertidal community composition. Specifically, we hypothesized that C. lingua …

Crystal Structure Of The Vibrio Cholerae Quorum-Sensing Regulatory Protein Hapr, Rukman S. De Silva, Gabriela Kovacikova, Wei Lin, Ronald K. Taylor, Karen Skorupski, F. Jon Kull

Dartmouth Scholarship

Quorum sensing in Vibrio cholerae involves signaling between two-component sensor protein kinases and the response regulator LuxO to control the expression of the master regulator HapR. HapR, in turn, plays a central role in regulating a number of important processes, such as virulence gene expression and biofilm formation. We have determined the crystal structure of HapR to 2.2-Å resolution. Its structure reveals a dimeric, two-domain molecule with an all-helical structure that is strongly conserved with members of the TetR family of transcriptional regulators. The N-terminal DNA-binding domain contains a helix-turn-helix DNA-binding motif and alteration of certain residues in this domain …

Clinical Presentation And Genotype Of Hepatitis Delta In Karachi, Tariq Moatter, Zaigham Abbas, Sabhita Shabir, Wasim Jafri

Department of Pathology and Laboratory Medicine

Aim:
To assess the clinical presentation and genotypes of delta hepatitis in local population.
Methods:
In this prospective study, 39 consecutive patients who were positive for HBsAg and hepatitis D virus (HDV) antibody were included. The patients were divided in two groups on the basis of presence or absence of HDV RNA and a comparative study was done. Genotype of HDV was determined in PCR positive patients.
Results:
Overall there is male dominance, in which 34 patients out of 39 (87.2%) were male. Twenty (51%) patients were from the adjacent areas of three provinces; Sindh, Punjab and Balochistan indicating the …

The Microbiological Analysis Of Composting, Marion Shepherd

All Theses

The objectives of this study were to: 1) perform a survey of South Carolina poultry farms to determine if the methods implemented resulted in the destruction of foodborne pathogens, and 2) determine the survival of E. coli O157:H7 in a dairy manure-based compost performed in uncontrolled environmental conditions.
In the survey of poultry farms Ninety-one (91) of 141 compost samples analyzed were positive for coliform populations ranging from 1.00 to 6.00 log10 CFU/g. Among finished compost samples (n=21), ca. 62%, 33%, and 14% were positive for coliforms, presumptive Salmonella, and presumptive Listeria, respectively.
In the investigation of the survival of …

The Nasal Cavity Is A Route For Prion Infection In Hamsters, Anthony E. Kincaid, Jason C. Bartz

Nebraska Center for Virology: Faculty Publications

Animals that naturally acquire the prion diseases have a well-developed olfactory sense that they utilize for a variety of basic behaviors. To assess the potential for the nasal cavity to serve as a point of entry for prion diseases, a small amount of prion-infected brain homogenate was placed inferior to the nostrils of hamsters, where it was immediately sniffed into the nasal cavity. Hamsters extranasally inoculated with the HY strain of transmissible mink encephalopathy (TME) agent had an incubation period that was not significantly different from per os inoculation of the same dose of the HY TME agent. However, the …

Conservation Of The Sos Operon, Umudc, In Acinetobacter Species, Ching-Li Lin

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science & Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Ching-Li Lin on May 1, 2007.

Impact Of Nutritional Supplements And Monosaccharides On Growth, Oxalate Accumulation, And Culture Ph By Sclerotinia Sclerotiorum, Steven L. Daniel, Bryan J. Culbertson, Norbert C. Furumo

Faculty Research & Creative Activity

Sclerotinia sclerotiorum D-E7 was studied to determine the impact of nutritional supplements and monosaccharides on growth, oxalate accumulation, and culture pH in broth media (initial pH c. 5). Cultures with 0.1% nutritional supplement (tryptone, yeast extract, or soytone) yielded minimal growth, 2–3 mM oxalate, and a final culture pH of 4.2–4.8. In contrast, cultures with 0.1% nutritional supplement and 25 mM glucose yielded significant growth, minimal oxalate (<1 mM), and a final culture pH of 2.8–3.7. Similar trends were observed when glucose in 0.1% soytone cultures was replaced with 25 mM d-mannose, l-arabinose, or d-xylose. With 1% soytone-25 mM glucose cultures, growth and oxalate accumulation (∼21 mM) occurred with little change in initial pH. This was not the case with 1% soytone-250 mM glucose cultures; increased glucose levels resulted in a decrease in oxalate accumulation (∼7 mM) and in final culture pH (3.4). Time-course studies with these cultures revealed that oxalate accumulation was suppressed during growth when the culture pH dropped to <4. Overall, these results indicate that (1) the decrease in external pH (i.e. acidification) was independent of oxalate accumulation and (2) acidification coupled to glucose-dependent growth regulated oxalate accumulation by Sclerotinia sclerotiorum.

Impact Of Nutritional Supplements And Monosaccharides On Growth, Oxalate Accumulation, And Culture Ph By Sclerotinia Sclerotiorum, Steven Daniel, Bryan Culbertson, Norbert Furumo

Faculty Research & Creative Activity

Sclerotinia sclerotiorum D-E7 was studied to determine the impact of nutritional supplements and monosaccharides on growth, oxalate accumulation, and culture pH in broth media (initial pH c. 5). Cultures with 0.1% nutritional supplement (tryptone, yeast extract, or soytone) yielded minimal growth, 2–3 mM oxalate, and a final culture pH of 4.2–4.8. In contrast, cultures with 0.1% nutritional supplement and 25 mM glucose yielded significant growth, minimal oxalate (<1 mM), and a final culture pH of 2.8–3.7. Similar trends were observed when glucose in 0.1% soytone cultures was replaced with 25 mM d-mannose, l-arabinose, or d-xylose. With 1% soytone-25 mM glucose cultures, growth and oxalate accumulation (∼21 mM) occurred with little change in initial pH. This was not the case with 1% soytone-250 mM glucose cultures; increased glucose levels resulted in a decrease in oxalate accumulation (∼7 mM) and in final culture pH (3.4). Time-course studies with these cultures revealed that oxalate accumulation was suppressed during growth when the culture pH dropped to <4. Overall, these results indicate that (1) the decrease in external pH (i.e. acidification) was independent of oxalate accumulation and (2) acidification coupled to glucose-dependent growth regulated oxalate accumulation by Sclerotinia sclerotiorum.

The Effect Of Radiation And Repeated Sub-Culturing On Tgf-Β1 Signaling In Frtl-5 Cells, Cheryl G. Burrell

Loma Linda University Electronic Theses, Dissertations & Projects

From our ongoing in vitro studies using the Fisher Rat Thyroid cell line-5 (FRTL-5) we recorded accelerated growth, reduced follicularization and reduction in thyroxin release that occurred as the cells were repeatedly sub-cultured. We also recorded that these changes occurred earlier and more rapidly following radiation exposure. We determined that TGF-β1 production increased under both conditions. We hypothesized that alteration in the TGF-β1 signaling pathway contributed to the changes observed in the cellular properties of FRTL-5 cells. Our objective was to examine some of the players in the TGF-β1 signaling pathway to determine whether radiation and/or repeated subculturing promoted changes …

Radiation-Induced Increases In Pkc Modulate Integrin Expression And Contribute To Fibrotic Changes, Pinal Rushikesh Pandya

Loma Linda University Electronic Theses, Dissertations & Projects

The sequences of events in fibrosis are similar to those in wound healing; however, the normal termination and resolution stages do not take place. The initial cellular response following ionizing radiation involves accumulation of the ECM (extracellular matrix) including collagen, fibronectin and the interaction of many growth factors (cytokines) with their receptors. There are a number of unanswered questions regarding many aspects of radiation-induced fibrosis, including the initial triggers and physical changes that initiate the process. We have confirmed that elevated pKC and other cellular changes following radiation are similar to those found at sites of inflammation. This information leads …

Downy Mildew In Vineyards, Diana Fisher, Andrew Taylor, Colin Gordon, Peter Magarey

Bulletins 4000 -

Downy mildew, caused by Plasmopara viticola, is a major fungal disease of grapevines that originates from North America. This disease was first detected in a commercial Western Australian vineyard in October 1998 and has since been found in most grape growing areas within the State. It is found in all other grape growing areas of Australia.

Plasmopara viticola is specific to grapevines (e.g. Vitis vinifera), although not all Vitis spp. are susceptible. The American rootstock species and hybrids are less susceptible or are resistant (refer to ‘Varietal susceptibility’). Other species of downy mildew, such as those found on …

Soil Microbial Community Response To Hexavalent Chromium In Planted And Unplanted Soil, Ioannis Ipsilantis, Mark S. Coyne

Plant and Soil Sciences Faculty Publications

Theories suggest that rapid microbial growth rates lead to quicker development of metal resistance. We tested these theories by adding hexavalent chromium [Cr(VI)] to soil, sowing Indian mustard (Brassica juncea), and comparing rhizosphere and bulk soil microbial community responses. Four weeks after the initial Cr(VI) application we measured Cr concentration, microbial biomass by fumigation extraction and soil extract ATP, tolerance to Cr and growth rates with tritiated thymidine incorporation, and performed community substrate use analysis with BIOLOG GN plates. Exchangeable Cr(VI) levels were very low, and therefore we assumed the Cr(VI) impact was transient. Microbial biomass was reduced …

Epidermoid Cyst Of The Ovary, Romana Idress, Zubair Ahmad, Khurrum Minhas, Naila Kayani

Department of Pathology and Laboratory Medicine

Epidermoid cysts of the ovary are rare lesions and are uniformly incidental findings in hysterectomy specimens. They must be differentiated from mature cystic teratomas of the ovary, and some authors believe them to be monodermal teratomas. The histogenesis is still uncertain. We report an epidermoid cyst of the ovary which was an incidental finding.

An Infection Model For Examining The Effects Of Gender And Diabetic State On Proinflammatory Cytokine Secretion By Phagocytic Cells In Response To Infection With Burkholderia Pseudomallei, Laura L. Dickey

Theses and Dissertations

Burkholderia pseudomallei is an opportunistic soil pathogen that causes melioidosis, a life-threatening human disease prevalent in Southeast Asia, northern Australia, the Middle East, Africa, and South America. The organism also causes disease in plants and animals. Persons with severe melioidosis usually die of septicemia. Relatively little is known regarding the virulence mechanisms of B. pseudomallei; however, several putative virulence determinants have been identified. The organism is able to invade and replicate within phagocytic cells and is particularly pathogenic in males with diabetes mellitus. B. thailandensis is closely related to B. pseudomallei, but is not pathogenic. This study examines various in …