Microbiology

Assessing the Source of Fecal Contamination in Streams on Kaua'i Based on Concentration and Genotypes of FRNA Bacteriophages

Vithanage, Gayatri — Mon, 01 Aug 2005 00:00:00 GMT

Extensive data from O'ahu indicate that all streams on this island consistently exceed the USEPA standards (200 fecal coliform/100 ml, 33 enterococci/100 ml) for water quality. Soil was determined to be the source of the elevated counts of these bacteria. In tropical areas, as Hawai'i, these bacteria are able to survive and multiply in the soil. Thus, these bacteria can end up in nearby streams after heavy rains or due to erosion. As a result, the USEPA recommended indicator bacteria (fecal coliform, enterococci) cannot be used to reliably determine when waters in tropical areas are fecally contaminated. Several alternative indicators have been proposed for such areas such as C. perfringens and FRNA coliphages. Extensive monitoring data does not exist for the other islands of Hawai'i. Kaua'i differs from O'ahu in that it is older, wetter and contains an abundance of cesspools. The Nawiliwili Watershed, on the island of Kaua'i, was chosen for this study. Sampling was conducted over a period of one year, and all samples were assayed for the traditional USEPA indicators (fecal, coliform, enterococci) as well as two alternative indicators (C. perfringens, FRNA coliphages). Of the 14 sites sampled, 12 contained levels of fecal coliform and enterococci that exceeded the USEPA standards (200 fecal coliform/100 ml and 33 enterococci/100 ml. This is similar to what has been documented in O'ahu streams. Based on the concentrations of these indicator bacteria, the USEPA would deem these sites as sewage contaminated. However, monitoring of these same sites for C. perfringens indicated that there was no sewage contamination (geometric mean values fell below the proposed standard of 50 CFU/100 ml). FRNA coliphage data indicate that cesspools may be leaching into nearby streams. Two streams (Nawiliwili, Papakōlea) had geometric mean levels greater than the 50 PFU/100 ml (based on O'ahu streams). Other streams in the watershed may be sporadically contaminated by cesspool because elevated FRNA coliphage levels were detected on occasion. Genotyping these FRNA coliphage isolates furthered supported the theory that cesspools were contaminating these sites because 98% of the FRNA isolates were typed as human while only 2% were typed as of animal origin. Current USEPA standards (fecal coliform, enterococci) are not reliable indicators of sewage pollution in tropical areas, thus, alternative indicators such as C. perfringens and FRNA coliphages may prove to be better indicators in these areas.

Are Fecal Sterols a Possible Alternative Indicator of Human Waste Contamination in Hawaiian Recreational Waters?

Brostrom, Kathleen A — Mon, 01 Aug 2005 00:00:00 GMT

Many of Hawaii’s recreational streams and beaches contain high fecal indicator bacteria levels that are not indicative of sewage pollution. Instead, this pollution is due to environmental sources of fecal bacteria which reside and multiply in tropical soils. Current EPA fecal indicator bacteria are no longer representative of human fecal contamination in tropical waters. Fecal sterols have been used as chemical indicators of fecal pollution in many parts of the world. The primary sterol found in human feces is coprostanol. Detection and quantification of coprostanol and related sterols using GCMS analysis provides a fingerprint that can be used to characterize fecal contamination. The objective of this study was to assay for fecal sterols as an independent method to determine whether streams in Hawaii are contaminated with sewage. This method was applied to ambient streams, a stream recently contaminated by a sewage spill, and a stream suspected to be affected by a sewage line leak. The results of this study showed that some ambient streams in Hawaii contain high levels of fecal indicator bacteria, but low concentrations of coprostanol (<10 ng/L). A stream contaminated with sewage during a sewage spill event contained high concentrations of coprostanol (18,000 ng/L) in the first 24 hours after contamination, but this level dropped to ≤ 60 mg/L after 72 hours. A stream suspected to be contaminated with sewage contained significant levels of coprostanol (>1000 ng/L) when fecal indicators were also high, confirming a possible sewage line leak. This study demonstrated that coprostanol is a useful and independent measurement of sewage pollution. It is best used in conjunction with other fecal indicators and human fecal markers if confirmation of human fecal pollution is sought.

Assessing the Persistence and Multiplication of Fecal Indicator Bacteria in Hawai'i Soil Environment

Byappananhalli, Muruleedhara N — Fri, 01 Dec 2000 00:00:00 GMT

Traditional fecal indicator bacteria such as fecal coliform, E.coli and enterococci have been shown to be unreliable indicators of the hygienic quality of recreational waters under tropical conditions. One of the major reasons for considering these bacteria as ineffective indicators of water quality in warm, tropical regions is that they are consistently found in natural environments (plants, soil, water) in the absence of any significant contamination of these environments. Since preliminary studies conducted in Hawaii had indicated soil as the major environmental source of elevated concentrations of these bacteria in environmental waters, the aim of this study was to focus on the soil environment to specifically address two assumptions made by regulatory agencies in using fecal bacteria as indicators of water quality: first, there should not be an environmental source of these indicator bacteria unrelated to sewage or fecal matter contamination, and second, the indicator bacteria do not multiply in the environment. To determine the validity of these two assumptions under tropical conditions in Hawaii and possibly other tropical locations, various experiments were conducted. The major findings are as follows. 1) Analysis of soil samples collected from various locations representing major soil groups on the island of Oahu showed that fecal indicator bacteria are naturally found in most of the soil environments, indicating that the fecal bacteria have adapted to the soil conditions to become part of soil biota. 2) Evidence was obtained to show that the soil contains adequate nutrients to sustain the populations of these bacteria. 3) Growth and multiplication of fecal indicator bacteria in natural soil was dependent on available nutrients (particularly carbon), moisture and competing microorganisms. In conclusion, tropical soil conditions are suboptimal for the multiplication of fecal indicator bacteria. Consequently, these bacteria in natural soil conditions will probably grow and multiply sporadically when conditions are relatively optimal. Although concentrations of fecal indicator bacteria in soil represent only a small fraction of the microbiota, their counts are significant enough in numbers not only to impact the quality of recreational waters but also to nullify two of the assumptions used in the application of recreational water quality standards. Thus, there is a need for an alternate and more reliable indicator of water quality in Hawaii and other tropical locations.

Effect of bioaugmentation and diesel fuel type on soil bioremediation

Chua-Chiaco, Barrie Wu — Sat, 01 Aug 1998 00:00:00 GMT

The enhancement of bioremediation by bioaugmentation in soil contaminated with diesel fuel No. 2 and No. 6 (Bunker C) is uncertain. A clayey soil was contaminated with 6,000 mg of either diesel fuel per kg of soil and seeded (5 x 10-7 cells/g of soil) with a Hawaii soil bacterium (UH138) known to utilize several hydrocarbons. The soil was limed, fertilized, and incubated in jars at 30°C for several months. The concentrations of total petroleum hydrocarbons (TPH) and of polycyclic aromatic hydrocarbons (PAH) in soil were measured by gravimetry and immunoassay, respectively. Poisoned controls (0.6% HgCl2) were used to determine the extent of hydrocarbon degradation due to microbial activity. A rapid first order biodegradation of TPH (84% in 23 days) occurred in soil contaminated with diesel fuel No. 2, regardless of bacterial seeding. Biodegradation of PAH was linear and reached 84% by day 98 in both seeded and unseeded treatments. Bioaugmentation had no effect on bioremediation of diesel fuel No.2. The decrease in TPH and PAH was paralleled by an increase in populations of total bacteria, phenanthrene-degrading bacteria and microorganisms capable of utilizing hexadecane and diesel fuel No. 2 as well as by an enhancement in CO2 evolution by the soil. Indigenous Zygomycetes grew profusely in diesel fuel No. 2 contaminated soil. Cunninghamella echinulata var. echinulata was isolated from the soil and was shown to be able to utilize several hydrocarbons. Thus, Zygomycetes may have contributed to the rapid decrease in contaminant. In soil contaminated with diesel fuel No. 6, the measurements of TPH and PAH were more variable due to the uneven distribution of the product. No biodegradation of the contaminant occurred over a period of 138 days. The growth of Zygomycetes was scant. The counts of total bacteria remained unchanged after the addition of diesel fuel No. 6. However, counts of the indigenous phenanthrene-degrading bacteria increases dramatically ( 4 log units) during the first 54 days whereas the level of the seeded bacteria remained stable. The counts of mineral oil degraders decrease by 2 log units after day 2. Co2 evolution from the soil confirmed that diesel fuel No. 6 was not degraded by either the indigenous microflora or the seeded bacterium. Thus, diesel fuel No. 2 was highly degradable by the indigenous population, however, diesel fuel No. 6 was recalcitrant. Thesis (M.S.)--University of Hawaii at Manoa, 1998.; Includes bibliographical references (leaves 106-117).; Available also on microfiche.

Characterization of genes involved in heterocyst differentiation and pattern formation in the cyanobacterium Anabaena sp. strain PCC 7120

Borthakur, Pritty B — Tue, 01 Jan 2008 00:00:00 GMT

Thesis (Ph.D.)--University of Hawaii at Manoa, 2008.; The goal of this research was to understand regulation of heterocyst differentiation in Anabaena sp. strain PCC 7120 by characterizing regulatory genes for heterocyst formation and their mutants. Anabaena is a filamentous cyanobacterium that forms specialized cells for nitrogen fixation, called heterocysts, which differentiate from vegetative cells at intervals of 10--12 cells. Two genes, patS and hetN, are known to suppress the differentiation of vegetative cells into heterocysts for establishing a de novo pattern and maintaining a pattern of heterocysts along the filament. A mutant, UHM100, was created to study the function of both genes by deleting patS and making expression of hetN conditional. This study has established that PatS and HetN are members of two separate heterocyst suppression pathways. In absence of nitrogen, inactivation of both patS and hetN increases heterocyst differentiation to nearly 100%, giving rise to a phenotype called 'multiple contiguous heterocysts' (Mch). UHM100 has an Mch phenotype even in the presence of combined nitrogen, which usually suppresses heterocyst differentiation. In absence of both patS and hetN, the expression of hetR, a master regulator of heterocyst differentiation, was observed in ∼55% cells and was asynchronous. The distribution of heterocysts next to a vegetative cell in UHM 100 was found to be nonrandom. These results suggest that besides PatS and HetN, there are other factors that influence pattern formation in Anabaena PCC 7120. A heterocyst-deficient (Hef) spontaneous mutant, NSM6, was isolated from UHM 100. A novel gene, alr9018, from the Anabaena Epsilon plasmid complemented NSM6 and restored the Mch phenotype of this mutant. Transconjugants of Anabaena PCC 7120 containing the cloned alr9018 gene fixed 50% more N2 than PCC 7120, suggesting that multiple copies of alr9018 enhance heterocyst development. This is the first report showing that the Epsilon plasmid of Anabaena PCC 7120 contains genes involved in heterocyst differentiation. Expression of alr9018 was observed in both vegetative cells and heterocysts. Similar to alr9018, hetR could also restore the Mch phenotype in NSM6, suggesting functional similarity between a1r9018 and hetR. The Alr9018 protein contains an NTPase domain, which is a characteristic of proteins involved in signal transduction.; Includes bibliographical references (leaves 89-97).; Also available by subscription via World Wide Web; 97 leaves, bound 29 cm

Bacterial profiles in healthy and Montipora white syndrome affected Montipora capitata mucus and the identification of potential etiologic agents

Smith, Ashley — Tue, 01 Jan 2008 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2008.; Includes bibliographical references (leaves 59-69).; ix, 87 leaves, bound ill. 29 cm

Research on the diversity and the biomedical potential of marine fungi in Hawaii

He, Mingxiao — Mon, 01 Jan 2007 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2007.; Includes bibliographical references (leaves 50-54).; x, 54 leaves, bound ill. 29 cm

Genes involved in nitrogen fixation and heterocyst differentiation in Anabaena sp. PCC 7120

Yamaura, Hiroshi — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 45-57).; vi, 57 leaves, bound ill. 29 cm

Genes involved in diazotrophic growth of Anabaena sp. PCC 7120

Nayar, Asha Sivasankaran — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 47-55).; vi, 55 leaves, bound ill. 29 cm

Kauai's potable groundwater sources : assessing its vulnerability to fecal contamination for the pending groundwater rule

Sato, Dayna — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 105-109).; ix, 109 leaves, bound col. ill., col. maps 29 cm

Impact of tropical plants on microbial activity and diversity in soil contaminated with petroleum hydrocarbons

Shibata, Alexandra Ku — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 138-163).; xii, 163 leaves, bound ill. 29 cm

Characterization of secretogranin III in mast cells

Prasad, Prerna — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 84-91).; ix, 91 leaves, bound ill. 29 cm

Investigating the regulation of fatty acid degradation in Pseudomonas aeruginosa

Nguyen, David Tran — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 143-160).; xx, 160 leaves, bound ill. 29 cm

Characterization of the Acyl-CoA binding domain containing 3 protein

Li, Zengqiu — Sun, 01 Jan 2006 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2006.; Includes bibliographical references (leaves 53-55).; ii, 55 leaves, bound ill. (some col.) 29 cm

Cytotoxicity and antiproliferative effects of extracts from coffee cherry fruit on cell lines from normal breast tissue and from non-invasive and invasive breast cancers

Meujo, Damaris Agathe — Sat, 01 Jan 2005 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2005.; Includes bibliographical references (leaves 106-109).; xxi, 109 leaves, bound ill. (some col.) 29 cm

The effects of acute infection and inflammation on phase II detoxifying enzymes

Welch, Darcy L — Mon, 01 Jan 2007 00:00:00 GMT

Thesis (M.S.)--University of Hawaii at Manoa, 2007.; Includes bibliographical references (leaves 71-82).; xiii, 82 leaves, bound 29 cm

Pseudomonas aeruginosa pathogenesis during cystic fibrosis lung infection and metabolism of lung surfactant component phosphatidylcholine

Son, Mike Seong — Tue, 01 Jan 2008 00:00:00 GMT

Includes supplementary digital materials.; Thesis (Ph.D.)--University of Hawaii at Manoa, 2008.; Pseudomonas aeruginosa, an opportunistic Gram-negative bacterium, is the principal cause of hospital-acquired pneumonia, and is responsible for the high morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa also causes a wide range of human diseases including otitis media, keratitis, endocarditis, osteochondritis, pyelonephritis, cellulites and septicemia. Despite the considerable effort expended toward studying P. aeruginosa infections and virulence expression, the pathogenesis of P. aeruginosa-associated diseases and the ability of P. aeruginosa to reach high cell density (HCD) during chronic lung infections in CF patients remains enigmatic. We hypothesized that in the lung environment, P. aeruginosa degrades the naturally occurring lung surfactant component phosphatidylcholine (PC), as a nutrient source to afford HCD replication and maintenance.; To test this central hypothesis, I have conducted microarray and real-time RT-PCR experiments directly on sputum samples from two chronically infected CF patients to obtain a snap shot of the metabolic profile of P. aeruginosa populations. The microarray and real-time RT-PCR data provide strong evidence substantiating our initial hypothesis that PC may serve as a nutrient source to afford HCD replication and maintenance.; Using bioinformatics and the microarray data, I was able to identify the PC degradative genes expressed during P. aeruginosa lung infection, including the fatty acid degradative (fad), choline metabolism (bet), and glycerol metabolism (glp) genes. The bet and glp pathways have been previously characterized, however, the fad pathway remains a mystery and can only be predicted based on the established E. coli fad pathway. Using this pathway as a model, I initiated investigations into deciphering this pathway in P. aeruginosa. I present here the characterization of the fadD genes (acyl-CoA synthetases), preliminary data on the characterization of the fadBA operons (acyl-CoA thiolases and acyl-CoA dehydrogenases), and data that link the association of the fad, bet, and glp pathways in regard to PC and PC component degradation.; Includes bibliographical references (leaves xxx-xxx).; Also available by subscription via World Wide Web; 293 leaves, bound 29 cm

Base composition of Mycoplasma DNA and the formation of specific DNA-DNA hybrids

Fox, Richard Howard — Sun, 01 Jan 1967 00:00:00 GMT

Typescript.; Thesis (Ph. D.)--University of Hawaii, 1967.; Bibliography: leaves 77-86.; ix, 86 l graphs, tables

Structural and genetic studies of chicken 7S immunoglobulin allotypes

Wakeland, Edward K — Thu, 01 Jan 1976 00:00:00 GMT

Typescript.; Thesis (Ph. D.)--University of Hawaii at Manoa, 1976.; Bibliography: leaves 141-150.; Microfiche.; xv, 150 leaves ill

Factors affecting 7S and 17S antibody concentrations and affinities in chickens

Yamaga, Karen — Tue, 01 Jan 1974 00:00:00 GMT

Typescript.; Thesis (Ph. D.)--University of Hawaii at Manoa, 1974.; Bibliography: leaves 131-141.; xiii, 141 leaves ill