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The response of macroinvertebrate and protozoan colonization dynamics to industrial pollution was determined in a thermal organic effluent and in a metals effluent. The relationship of the time course data to the MacArthur-Wilson model is considered, along with Pinkham and Pearson's community similarity index and traditional community structural parameters. The response of both communities is similar with respect to equilibrium number of taxa at both sites; however, the fitted rate constant, G, was accelerated in the organic effluent for the protozoans. The macroinvertebrate G in the same effluent showed a dose response. There were no differences in rate between stations in the metals effluent for either the protozoans or the macroinvertebrates, although the equilibrium number of taxa was decreased downstream. Community similarity indices shows similar macroinvertebrate and protozoan communities at all three stations in the organic system at equilibrium, although some differences occur in the early stages of colonization. The communities in the metals effluent are most often significantly different at the downstream station, which was the most impacted. Community diversity indices indicate the same results for the equilibrium community as do the other methods of evaluation. While community colonization studies have been suggested as a method of water quality assessment, colonization dynamics and particularly the fitted rate constant, G, may be misleading unless considered in conjunction with equilibrium information. / Master of Science

Neste estudo foram isolados e caracterizados flagelados de 20 amostras de solo e água do Brasil classificados no gênero Euglena por parâmetros taxonômicos tradicionais, e distribuídos em 5 grupos de acordo com características morfológicas. Análises filogenéticas do gene SSUrDNA das 20 amostras resultou em 25 seqüências posicionadas nos grupos A2 e A3 do gênero Euglena definidos em estudos anteriores, separando as seqüências obtidas em 7 clados distintos, revelando uma maior diversidade que a obtida por parâmetros taxonômicos tradicionais. A fim de resolver os relacionamentos intra-específicos do grupo A3, realizamos uma análise filogenética restrita deste grupo, que apresentou árvores congruentes e subclados bem suportados. Representantes de cada espécie foram analisados por microscopia eletrônica de varredura, revelando dois padrões principais de estrias e poros da película. Os isolados caracterizados neste estudo são os primeiros do Brasil estabelecidos em cultura, validados por análises filogenéticas e morfologicamente caracterizados. / In this study we isolated and characterized flagellates from 20 soil and water samples of Brazil, classified in the genus Euglena based on traditional taxonomic parameters and distributed in 5 groups according to morphological characteristics. Phylogenetic analyses of the SSUrDNA of the 20 samples resulted in 25 sequences that where positioned in previously defined groups A2 and A3 of the genus Euglena, separating the obtained sequences in 7 distinct clades, revealing a larger diversity than defined by traditional taxonomic parameters. To better resolve the intra-specific relationships of group A3, we conducted a phylogenetic analyses restricted to this group, generating congruent trees and well supported sub-clades. A representative of each species was selected and analyzed by scanning electron microscopy, revealing two major patterns of strip and pellicle pores. The isolates characterized in this study are the first in Brazil to be established in culture, validated by phylogenetic analyses and morphologically characterized.

<p>Циљ истраживања је био да се утврди појава и преваленца паразита Marteilia<br />refringens у медитеранској дагњи (Mytilus galloprovincialis) гајеној у<br />Бококоторском заливу. Осим тога имали смо за циљ да утврдимо да ли<br />еколошки фактори (температура морске воде, салинитет, концентрација<br />кисеоника, pH вредност) и присуство бактерија (Escherichia coli и стрептококи<br />фекалног порекла) утичу на појаву овог паразита у медитеранској дагњи.<br />Вршили смо испитивања на шест различитих локација &ndash; узгајалишта<br />медитеранске дагње у Бококоторском заливу. Укупно је узорковано 960 јединки<br />медитеранске дагње. Marteilia refringens је утврђена на четири локације -<br />узгајалишта. Укупна преваленца паразита Marteilia refringens је износила<br />1,25%. Кретала се од 0% на две локације, 0,56% на једној, 1,67% на једној<br />локацији до максимално 3,33% на двe локације. Пратили смо евентуалну<br />појаву паразита током дванаестомесечног периода, а утврдили смо његово<br />присуство од септембра до децембра и у марту месецу. Од праћених<br />еколошких фактора утврдили смо да највећи утицај на појаву паразита има pH<br />вредност морске воде, с тим што се Marteilia refringens појављује при нижој pH<br />вредности од просечне. Утврдили смо и утицај концентрације кисеоника у<br />морској води, при чему се Marteilia refringens јављала при вишим<br />концентрацијама кисеоника. Нисмо утврдили утицај температуре на појаву<br />паразита, али смо забележили појаву паразита при минималној температури<br />од 11,2˚C. Нисмо статистички доказали утицај салинитета и појаве бактерија у<br />морској води на појаву Marteiliae refringens. Доказали смо негативан утицај<br />појаве паразита Marteilia refringens на индекс кондиције медитеранске дагње.<br />Све мартелије утврђене цитолошким и хистолошким испитивањима смо<br />потврдили и молекуларним испитивањима, при чему смо утвдили да наши<br />изолати паразита припадају М типу Marteiliae refringens. Сви н</p> / <p>Cilj istraživanja je bio da se utvrdi pojava i prevalenca parazita Marteilia<br />refringens u mediteranskoj dagnji (Mytilus galloprovincialis) gajenoj u<br />Bokokotorskom zalivu. Osim toga imali smo za cilj da utvrdimo da li<br />ekološki faktori (temperatura morske vode, salinitet, koncentracija<br />kiseonika, pH vrednost) i prisustvo bakterija (Escherichia coli i streptokoki<br />fekalnog porekla) utiču na pojavu ovog parazita u mediteranskoj dagnji.<br />Vršili smo ispitivanja na šest različitih lokacija &ndash; uzgajališta<br />mediteranske dagnje u Bokokotorskom zalivu. Ukupno je uzorkovano 960 jedinki<br />mediteranske dagnje. Marteilia refringens je utvrđena na četiri lokacije -<br />uzgajališta. Ukupna prevalenca parazita Marteilia refringens je iznosila<br />1,25%. Kretala se od 0% na dve lokacije, 0,56% na jednoj, 1,67% na jednoj<br />lokaciji do maksimalno 3,33% na dve lokacije. Pratili smo eventualnu<br />pojavu parazita tokom dvanaestomesečnog perioda, a utvrdili smo njegovo<br />prisustvo od septembra do decembra i u martu mesecu. Od praćenih<br />ekoloških faktora utvrdili smo da najveći uticaj na pojavu parazita ima pH<br />vrednost morske vode, s tim što se Marteilia refringens pojavljuje pri nižoj pH<br />vrednosti od prosečne. Utvrdili smo i uticaj koncentracije kiseonika u<br />morskoj vodi, pri čemu se Marteilia refringens javljala pri višim<br />koncentracijama kiseonika. Nismo utvrdili uticaj temperature na pojavu<br />parazita, ali smo zabeležili pojavu parazita pri minimalnoj temperaturi<br />od 11,2˚C. Nismo statistički dokazali uticaj saliniteta i pojave bakterija u<br />morskoj vodi na pojavu Marteiliae refringens. Dokazali smo negativan uticaj<br />pojave parazita Marteilia refringens na indeks kondicije mediteranske dagnje.<br />Sve martelije utvrđene citološkim i histološkim ispitivanjima smo<br />potvrdili i molekularnim ispitivanjima, pri čemu smo utvdili da naši<br />izolati parazita pripadaju M tipu Marteiliae refringens. Svi n</p> / <p>The aim of the research was to determine the presence and prevalence of the<br />parasite Marteilia refringens in Mediterranean mussels (Mytilus galloprovincialis)<br />cultivated in Boka kotorska Bay. In addition we aimed to determine wheather<br />environmental factors (temperature of seawater, salinity, oxygen concentration, pH<br />value) and the presence of bacteria (Escherichia coli and streptococci of faecal<br />origin) have influence on the occurrence of this parasite in the Mediterranean<br />mussels. We conducted tests at six different locations. In total 960 individuals of<br />Mediterranean mussels were sampled and included in the study. Marteilia refringens<br />was found at four locations. The overall prevalence of the parasite Marteilia<br />refringens was 1.25%. It ranged from 0% at two locations, 0,56% at one, 1.67% at<br />one to 3,33% at two locations. We traced the possible occurrence of the parasite<br />during a twelve month period, and we found its presence from September till<br />December and in March. We found that the pH value of seawater had the greatest<br />impact on the occurence of the parasite from all monitored environmental factors.<br />Marteilia refringens occurs at lower pH values than the average measured pH<br />values of seawater. We also found that Marteilia refringens occured at the higher<br />concentrations of oxygen in the seawater. We did not determine the effect of<br />temperature on the occurrence of the parasite, but we recorded the occurrence of<br />this parasite at a minimum temperature of 11,2˚C. We did not prove that salinity and<br />the presents of bacteria had an influence on the occurrence of Marteilia refringens.<br />We determined a negative impact of Marteilia refringens on the condition index of<br />Mediterranean mussel. All parasites, determined by cytologic and histologic<br />examination were confirmed by molecular methods. We determined that all our<br />isolates belonged to the M type of Marteilia refringens. All our isolates show genetic<br />uniformity, but also show genetic similarity with other isolates of Marteilia</p>

Um gene que codifica uma piruvato/indolpiruvato descarboxilase (PDC/IPDC) está presente no tripanossomatídeo de plantas Phytomonas serpens. A PDC atua na fermentação alcoólica, enquanto que a IPDC atua na biossíntese do fitormônio ácido indol-3-acético (AIA). Análises filogenéticas indicam que a PDC/IPDC de P. serpens é monofilética com IPDCs de gama-proteobactérias, sugerindo um evento de transferência horizontal gênica. A análise de meios de cultura de P. serpens confirma a produção de etanol e AIA. A funcionalidade do fitormônio foi confirmada em ensaios de alongamento de hipocótilos de tomateiros. Tomates inoculados com P. serpens mostraram aumento no teor de AIA-amida e -éster conjugados. A atividade PDC foi mostrada em extratos de P. serpens. Concluímos que a PDC/IPDC seria uma 2-cetoácido descaboxilase com atividade catalítica variável para diferentes substratos. A atividade PDC parece ser predominante em P. serpens, representando um mecanismo para oxidar parte do NADH formado na glicólise, principal responsável pela produção de ATP neste organismo. / A gene codifying a pyruvate/indolepyruvate decarboxylase (PDC/IPDC) is present in the plant trypanosomatid Phytomonas serpens. PDC acts in the alcoholic fermentation, whyle IPDC acts in the biosynthesis of the phytohormone indole-3-acetic acid (IAA). Phylogenetic analysis indicate that P. serpens PDC/IPDC is monophyletic with gamma-proteobacteria IPDCs, suggesting a horizontal gene transfer event. Analysis of P. serpens culture media confirms production of ethanol and IAA. The functionality of the phytohormone was confirmed by tomato hypocotyl elongation tests. Tomatoes inoculated with P. serpens showed an increase in the concentration of IAA amide and ester conjugated. PDC activity was shown in P. serpens extracts. We conclude that the PDC/IPDC would be a 2-keto acid decaboxylase with variable catalytic activity for different substrates. The PDC activity appears to be prevalent in P. serpens representing a mechanism to oxidize part of NADH formed in glycolysis, responsible for ATP production in this organism.

Neste estudo foram isolados e caracterizados flagelados de 20 amostras de solo e água do Brasil classificados no gênero Euglena por parâmetros taxonômicos tradicionais, e distribuídos em 5 grupos de acordo com características morfológicas. Análises filogenéticas do gene SSUrDNA das 20 amostras resultou em 25 seqüências posicionadas nos grupos A2 e A3 do gênero Euglena definidos em estudos anteriores, separando as seqüências obtidas em 7 clados distintos, revelando uma maior diversidade que a obtida por parâmetros taxonômicos tradicionais. A fim de resolver os relacionamentos intra-específicos do grupo A3, realizamos uma análise filogenética restrita deste grupo, que apresentou árvores congruentes e subclados bem suportados. Representantes de cada espécie foram analisados por microscopia eletrônica de varredura, revelando dois padrões principais de estrias e poros da película. Os isolados caracterizados neste estudo são os primeiros do Brasil estabelecidos em cultura, validados por análises filogenéticas e morfologicamente caracterizados. / In this study we isolated and characterized flagellates from 20 soil and water samples of Brazil, classified in the genus Euglena based on traditional taxonomic parameters and distributed in 5 groups according to morphological characteristics. Phylogenetic analyses of the SSUrDNA of the 20 samples resulted in 25 sequences that where positioned in previously defined groups A2 and A3 of the genus Euglena, separating the obtained sequences in 7 distinct clades, revealing a larger diversity than defined by traditional taxonomic parameters. To better resolve the intra-specific relationships of group A3, we conducted a phylogenetic analyses restricted to this group, generating congruent trees and well supported sub-clades. A representative of each species was selected and analyzed by scanning electron microscopy, revealing two major patterns of strip and pellicle pores. The isolates characterized in this study are the first in Brazil to be established in culture, validated by phylogenetic analyses and morphologically characterized.

Um gene que codifica uma piruvato/indolpiruvato descarboxilase (PDC/IPDC) está presente no tripanossomatídeo de plantas Phytomonas serpens. A PDC atua na fermentação alcoólica, enquanto que a IPDC atua na biossíntese do fitormônio ácido indol-3-acético (AIA). Análises filogenéticas indicam que a PDC/IPDC de P. serpens é monofilética com IPDCs de gama-proteobactérias, sugerindo um evento de transferência horizontal gênica. A análise de meios de cultura de P. serpens confirma a produção de etanol e AIA. A funcionalidade do fitormônio foi confirmada em ensaios de alongamento de hipocótilos de tomateiros. Tomates inoculados com P. serpens mostraram aumento no teor de AIA-amida e -éster conjugados. A atividade PDC foi mostrada em extratos de P. serpens. Concluímos que a PDC/IPDC seria uma 2-cetoácido descaboxilase com atividade catalítica variável para diferentes substratos. A atividade PDC parece ser predominante em P. serpens, representando um mecanismo para oxidar parte do NADH formado na glicólise, principal responsável pela produção de ATP neste organismo. / A gene codifying a pyruvate/indolepyruvate decarboxylase (PDC/IPDC) is present in the plant trypanosomatid Phytomonas serpens. PDC acts in the alcoholic fermentation, whyle IPDC acts in the biosynthesis of the phytohormone indole-3-acetic acid (IAA). Phylogenetic analysis indicate that P. serpens PDC/IPDC is monophyletic with gamma-proteobacteria IPDCs, suggesting a horizontal gene transfer event. Analysis of P. serpens culture media confirms production of ethanol and IAA. The functionality of the phytohormone was confirmed by tomato hypocotyl elongation tests. Tomatoes inoculated with P. serpens showed an increase in the concentration of IAA amide and ester conjugated. PDC activity was shown in P. serpens extracts. We conclude that the PDC/IPDC would be a 2-keto acid decaboxylase with variable catalytic activity for different substrates. The PDC activity appears to be prevalent in P. serpens representing a mechanism to oxidize part of NADH formed in glycolysis, responsible for ATP production in this organism.

LD2668 .T4 1941 P41 / Master of Science

The aim of this thesis was to provide a picture of the trypanosomosis and drug resistance prevalence in Eastern Province of Zambia, to understand the underlying factors of drug resistance (drug use habits), to improve the diagnosis of trypanosomosis in livestock and finally, to improve the diagnosis of isometamidium resistance in T.congolense. After an introductory part where available trypanosomosis and trypanocide resistance diagnostic methods are described and discussed, the body of the thesis is divided in two main sections. In the first section are presented the results of a cross-sectional and a longitudinal epidemiological survey describing the geographical distribution of trypanosomosis cases, of resistant isolates and of cattle treated with isometamidium chloride. The results of the monitoring of unsupervised treatments of cattle with isometamidium by farmers and veterinary assistants with the Isometamidium-ELISA technique are also presented. The second section describes the development of two new diagnostic methods, the first one allowing the diagnosis of trypanosome infections with high sensitivity and specificity through semi-nested polymerase chain reaction and restriction fragment length polymorphism. This is the first report of a pan-trypanosome PCR test (a single PCR test for the diagnosis of all important pathogenic trypanosomes of cattle). The second new method that was developed allows the diagnosis of isometamidium resistant T.congolense strains by PCR-RFLP. This is the first report of a PCR based diagnostic test of trypanocide resistance in T. congolense.