Sida rhombifolia L belongs to the family Malvaceae, it is native to the American continent and can be found in all regions of Brazil. It is popularly known as prickly sida , and used in traditional medicine as anti-inflammatory, antidiabetic and lipid lowering. The aim of this study was to determine and evaluate the physical and chemical parameters of quality in the four seasons and the aerial parts and roots of the plant drug; determining the content of total polyphenols, flavonoids and tannins in the extracts of the aerial parts and roots of S. rhombifolia in four seasons; evaluating the antimicrobial and antioxidant activity in these same extracts; investigating the hypoglycemic and lipid lowering activity of extracts that have a higher content of flavonoids, as well as to analyze biochemical parameters of liver, kidney and pancreatic toxicity and cytotoxicity in cells NCTC clone 929 by incorporation of neutral red dye. The drug was collected at the four seasons in Santa Maria, RS, separated into aerial parts and roots and made hydroethanolic extracts 70% from each of these collections. The physico-chemical parameters varied according to the plant part analyzed and the season. The values found for strange matter, determination of water and ash insoluble in hydrochloric acid are consistent with those established for herbs. The winter collection showed higher content of total ash, both to shoots (21.20%) and roots (17.63%). The swelling rates varied significantly among the aerial parts and roots, and the highest rates obtained for each of the samples were collected in summer (35.47%) and autumn (18.40%), respectively. There were significant differences between the content of total polyphenols and flavonoids throughout the year and these have focused differently from the aerial parts and roots. The content of tannins and did not suffer seasonal variation nor the distribution between the parties analyzed the plant. The extracts of the aerial parts (172.50 g / mL) and roots (374.08 g / mL) of winter presented the best antioxidant activity. Extracts of S. rhombifolia showed good antibacterial activity while showed no activity against fungi. The extracts of the aerial parts and roots of the summer of the winter showed no hypoglycemic and lipid lowering activity in healthy animals, the markers of liver, kidney and pancreatic damage did not vary in relation to the control group, and the IC50 obtained at the cytotoxicity assays were 8.88 mg/mL e 12.57 mg/mL, respectively. / Sida rhombifolia L. pertence à família Malvaceae, é nativa do continente americano e pode ser encontrada em todas as regiões do Brasil. É conhecida popularmente por guanxuma, e utilizada na medicina tradicional como antiinflamatória, hipoglicemiante e hipolipidemiante. O objetivo deste trabalho foi determinar e avaliar os parâmetros físico-químicos de qualidade nas quatro estações do ano e nas partes aéreas e raízes da droga vegetal; realizar o doseamento de polifenóis totais, flavonoides e taninos condensados nos extratos das partes aéreas e raízes de S. rhombifolia nas quatro estações; avaliar a atividade antioxidante e antimicrobiana nestes mesmos extratos; investigar a atividade hipoglicemiante e hipolipidemiante dos extratos que apresentarem maior conteúdo de flavonoides, assim como analisar parâmetros bioquímicos de toxicidade hepática, renal e pancreática e citotoxicidade em células NCTC clone 929 por incorporação do corante vermelho neutro. A droga vegetal foi coletada nas quatro estações do ano no município de Santa Maria, RS, separada em partes aéreas e raízes e feitos extratos hidroetanólicos 70% de cada uma destas coletas. Os parâmetros físico-químicos variaram de acordo com a parte da planta analisada e a estação. Os valores encontrados para matéria estranha, determinação de água e cinzas insolúveis em ácido clorídrico estão de acordo com os estabelecidos para drogas vegetais. A coleta de inverno apresentou maior conteúdo de cinzas totais, tanto para as partes aéreas (21,20%) quanto para as raízes (17,63%). Os índices de intumescência variaram significativamente entre as partes aéreas e raízes, e os maiores índices obtidos para cada uma das coletas foram nas amostras do verão (35,47%) e outono (18,40%), respectivamente. Houve diferença significativa entre os conteúdos de polifenóis totais e flavonoides ao longo do ano e estes concentraram se diferentemente entre as partes aéreas e raízes. O conteúdo de taninos não sofreu variação entre as partes analisadas da planta e de sazonalidade. Os extratos das partes aéreas (172,50 μg/mL) e das raízes (374,08 μg/mL) do inverno foram os que apresentaram melhor atividade antioxidante. Os extratos de S. rhombifolia apresentaram boa atividade antibacteriana e não apresentaram atividade contra fungos. Os extratos das partes aéreas do verão e das raízes do inverno não demonstraram atividade hipoglicemiante e hipolipidemiante em animais sadios. Os marcadores de danos hepáticos, renais e pancreáticos não variaram em relação ao grupo controle, e as IC50 obtidas nos testes de citotoxicidade das partes aéreas e raízes foram 8,88 mg/mL e 12,57 mg/mL, respectivamente.

>Magister Scientiae - MSc / The water quality in many Cape Town Rivers and streams is a major challenge. Kuils River is subject to multiple land use impacts from upstream to downstream because of rapid urbanization in its catchment area. The main pollution sources are urban and industrial, organic matter from litter under the road-bridge, and golf course. However no systematic efforts have been made to evaluate and improve the health of the river in term of management. To assess impacts on water quality, this study was conducted from 4th September to 27th November 2012 in 5 selected sites in the upper reach of the Kuils river. The main aim was to compare the health of the river in 2012 with that found in 2005 using physical and chemical characteristics and the South Africa Scoring System (SASS). The statistical analysis showed a significant difference between and within sites. The water temperature, pH, dissolved oxygen concentration, total dissolved solids (TDS), and salinity were collected in situ by YSI 30 meter. To evaluate nutrient (nitrate and phosphorus) concentrations water samples were analyzed at UWC laboratory using spectrophotometer. In addition human activities, basic conditions (7.13 to 8.76), high total dissolved solids (416 to to 916.5 mg L¯¹) and salinity (0.31 to 0.71 mg L¯¹) concentrations were influenced by Malmesbury shales. Nitrate (0.1 to 3.1 mg L¯¹) and phosphorus (0.11 to 5.27 mg L¯¹) concentrations and the decrease in dissolved oxygen in November 2012 showed eutrophic conditions of the river. In the tributary site phosphorus (1.32 to 3.62 mg L¯¹) concentrations revealed hypertrophic condition compared to South Africa guideline. Macroinvertebrates sampled showed a total of 28 taxa grouped in 11 orders were sampled. Poor habitat diversity and water quality degradation were principal causes of low species diversity. The South Africa Score System version 5 (SASS5) and Average Score per Taxon (ASPT) indicated that the river is seriously impacted in 2012 compared to 2005 where water quality was in poor condition. The SASS and the ASPT scores were less than 50 and 4.2 at all sampling sites in most part of sampling period.

<p>Sa&nbsp; jedne&nbsp; strane&nbsp; se&nbsp; eksploatacija&nbsp; &scaron;ljunka&nbsp; navodi kao&nbsp; značajan&nbsp; ugrožavajući&nbsp; faktor&nbsp; sa&nbsp; velikim negativnim&nbsp; uticajem&nbsp; na&nbsp; vodena&nbsp; stani&scaron;ta&nbsp; i biodiverzitet,&nbsp; dok&nbsp; same&nbsp; &scaron;ljunkare&nbsp; mogu predstavljati&nbsp; vredne&nbsp; refugijume&nbsp; akavtičnog&nbsp; biodiverziteta.&nbsp; Osnovni&nbsp; cilj&nbsp; disertacije&nbsp; je određivanje&nbsp; najznačajnijih&nbsp; i&nbsp; relevantnih hidromorfolo&scaron;kih&nbsp; parametara&nbsp; koji&nbsp; utiču&nbsp; na strukturiranje&nbsp; makrofitskih&nbsp; zajednica&nbsp; u &scaron;ljunkarama&nbsp; duž&nbsp; plavnog&nbsp; područja&nbsp; reke&nbsp; Drine&nbsp; i određivanje&nbsp; korelacije&nbsp; između&nbsp; izdvojenih parametara&nbsp; i&nbsp; kvantitativnih&nbsp; indeksa&nbsp; makrofita.Istraživanje&nbsp; je&nbsp; vr&scaron;eno&nbsp; u&nbsp; toku&nbsp; letnjih&nbsp; meseci 2015, 2016, 2017 i 2018. godine na 18 &scaron;ljunkara (60 istraživačkih vegetacijskih sektora) u okviru tri&nbsp;&nbsp; eksploataciona&nbsp; polja&nbsp; u&nbsp; Crnoj&nbsp; Bari, Badovincima&nbsp; i&nbsp; Lipničkom&nbsp; &Scaron;oru&nbsp; i&nbsp; na&nbsp; četiri prirodna&nbsp; fluvijalna&nbsp; jezera&nbsp; (13&nbsp; istraživačkih vegetacijskih&nbsp; sektora)&nbsp; u&nbsp; plavnom&nbsp; području&nbsp; reke Drine.&nbsp; Makrofitska&nbsp; vegetacija&nbsp; je&nbsp; konstatovana na svih 18 istraţivanih &scaron;ljunkara, prikupljenih na tri eksploataciona polja (Badovinci, Crna Bara&nbsp; i<br />Lipniĉki &Scaron;or). Zabeležena je 31 biljna vrsta. Kao najučestalije,&nbsp; sa&nbsp; najvećom&nbsp; apsolutnom pokrovno&scaron;ću izdvojile su se vrste:&nbsp;<em> Potamogeton nodosus</em>&nbsp; Poiret,&nbsp; <em>Ceratophyllum&nbsp; demersum&nbsp; L subsp.&nbsp; demersum,&nbsp; Myriophyllum&nbsp; spicatum&nbsp; L,Najas&nbsp; marina&nbsp; L&nbsp; i&nbsp;&nbsp; Chara&nbsp; globularis&nbsp; Thuill&nbsp;</em> Na četiri prirodna fluvijalna jezera zabeleženo je 13 vrsta.&nbsp; Vrste&nbsp; <em>Vallisneria&nbsp; spiralis&nbsp; L,&nbsp; Elodea canadensis&nbsp; Michx,&nbsp; Callitriche&nbsp; palustris&nbsp; L, Potamogeton&nbsp; natans&nbsp; L&nbsp; i&nbsp; Nuphar&nbsp; lutea&nbsp; (L)&nbsp;</em> Sm izdvojile&nbsp; su&nbsp; se&nbsp; kao&nbsp; konstantne&nbsp; i&nbsp; dominantne. Vrednosti svih kvantitativnih indeksa makrofita,<br />značajno&nbsp; su&nbsp; veće&nbsp; za&nbsp; &scaron;ljunkare&nbsp; u&nbsp; poređenju&nbsp; sa prirodnim&nbsp; fluvijalnim&nbsp; jezerima&nbsp; na&nbsp; nivouLEAFPACS&nbsp; sektora.&nbsp; Na&nbsp; istraživanim &scaron;ljunkarama,&nbsp; analizom&nbsp; klasterovanja&nbsp; je<br />izdvojeno&nbsp; 13&nbsp; vegetacijskih&nbsp; grupa&nbsp; (VG):&nbsp; VG1<em> Ceratophyllum&nbsp; demersum</em>,&nbsp; VG2&nbsp; <em>Ceratophyllum demersum&nbsp; -&nbsp; Valisneria&nbsp; spiralis</em>,&nbsp; VG3&nbsp; <em>Chara contraria,</em>&nbsp; VG4&nbsp; <em>Chara</em>&nbsp; <em>globularis,</em>&nbsp; VG5&nbsp; <em>Elodea canadensis,</em>&nbsp; VG6&nbsp; <em>Elodea&nbsp; nuttallii</em>,&nbsp; VG7&nbsp; <em>Najas marina</em>,&nbsp; VG8&nbsp; <em>Najas&nbsp; minor,</em>&nbsp; VG9&nbsp; <em>Nitellopsis obtusa</em>,&nbsp; VG10&nbsp;<em> Nuphar&nbsp; lutea</em>,&nbsp; VG11 <em>Potamogeton&nbsp; nodosus</em>,&nbsp; VG12&nbsp;<em> Potamogeton natans&nbsp;</em> i&nbsp;&nbsp; VG13&nbsp; <em>Potamogeton&nbsp; pectinatus</em>.&nbsp; Na prirodnim&nbsp; fluvijalnim&nbsp; jezerima&nbsp; konstatovane&nbsp; su četiri&nbsp; vegetacijske&nbsp; grupe:&nbsp; VG5&nbsp;<em> Elodea canadensis</em>,&nbsp; VG10&nbsp; <em>Nuphar&nbsp; lutea,</em>&nbsp; VG12<em> Potamogeton natans&nbsp;</em> i&nbsp; VG14&nbsp; <em>Typha latifolia</em>.&nbsp; Na osnovu izmerenih fizičko-hemijskih parametara,kvalitet vode u većini &scaron;ljunkara odgovara II klasi kvaliteta, na osnovu čega se mogu okarakterisati kao&nbsp; vodna&nbsp; tela&nbsp; sa&nbsp; dobrim&nbsp; i&nbsp; boljim&nbsp; ekolo&scaron;kim potencijalom. Sve &scaron;ljunkare i fluvijalna jezera se klasifikuju&nbsp; kao&nbsp; visoko&nbsp; alkalna.&nbsp; Kvalitet&nbsp; vode&nbsp; u prirodnim&nbsp; fluvijalnim&nbsp; jezerima&nbsp; odgovara&nbsp; III&nbsp; &ndash; IV&nbsp; klasi&nbsp; kvaliteta&nbsp; voda,&nbsp; pri&nbsp; čemu&nbsp; se&nbsp; mogu okarakterisati&nbsp; kao&nbsp; vodna&nbsp; tela&nbsp; sa&nbsp; slabim&nbsp; do umerenim&nbsp; ekolo&scaron;kim&nbsp; statusom.&nbsp; Značajno&nbsp; veće<br />vrednosti&nbsp; ukupnih&nbsp; suspendovanih&nbsp; materija, hemijske&nbsp; i&nbsp; biolo&scaron;ke&nbsp; potro&scaron;nje&nbsp; kiseonika, ukupnog&nbsp; organskog&nbsp; kiseonika&nbsp; i&nbsp; nitrata zabeležene su na prirodnim fluvijalnim jezerima u poređenju sa &scaron;ljunkarama. Izmerene vrednosti fizičko-hemijskih&nbsp; parametara&nbsp; ukazuju&nbsp; na mezotrofni&nbsp; karakter&nbsp; lokaliteta&nbsp; u&nbsp; Badovincima&nbsp; i mezo-eutrofni&nbsp; karakter&nbsp; lokaliteta&nbsp; u&nbsp; Lipničkom &Scaron;oru,&nbsp; dok&nbsp; se&nbsp; &scaron;ljunkare&nbsp; na&nbsp; teritoriji&nbsp; Crne&nbsp; Bare mogu&nbsp; okarakterisati&nbsp; kao&nbsp; eutrofna&nbsp; jezera.Vrednosti&nbsp; LHMS&nbsp; (modifikacionog)&nbsp; skora&nbsp; za &scaron;ljunkare kretale su se u rasponu od 9&nbsp; &ndash;&nbsp; 15, dok su vrednosti LHQA skora (stani&scaron;nog diverziteta) bile u rasponu izmeĊu 33&nbsp; &ndash;&nbsp; 44. Sliĉne vrednosti za&nbsp; LHQA&nbsp; skor&nbsp; su&nbsp; izraĉunate&nbsp; i&nbsp; za&nbsp; prirodna fluvijalna&nbsp; jezera&nbsp; (36&nbsp; &ndash;&nbsp; 49).&nbsp; MeĊutim,&nbsp; vrednosti<br />LHMS&nbsp; skora&nbsp; za&nbsp; prirodna&nbsp; fluvijalna&nbsp; jezera&nbsp; su znaĉajno&nbsp; veće&nbsp; u&nbsp; odnosu&nbsp; na&nbsp; vrednosti&nbsp; LHMS skora za &scaron;ljunkare. Ovi podaci ukazuju na manje prisustvo&nbsp; antropogenog&nbsp; pritiska&nbsp; na&nbsp; &scaron;ljunkarama u poređenju sa prirodnim fluvijalnim jezerima uistraţivanom&nbsp; podruĉju.&nbsp; Fizičko -hemijski&nbsp; i hidromorfolo&scaron;ki&nbsp; parametri&nbsp; zajedno&nbsp; su&nbsp; objasnili 57.07&nbsp; %&nbsp; od&nbsp; ukupne&nbsp; varijanse&nbsp; vegetacijskih podataka,&nbsp; sa&nbsp; 16.57&nbsp; %&nbsp; deljenog&nbsp; efekta.&nbsp; Fizičkohemijski&nbsp; parametri&nbsp; kvaliteta&nbsp; vode&nbsp; objasnili&nbsp; su 17.02&nbsp; %&nbsp; varijabilnosti&nbsp; u&nbsp; strukturi&nbsp; makrofitske vegetacije.&nbsp; Kao&nbsp; najsignifikantniji&nbsp; parametri<br />izdvojili&nbsp; su&nbsp; se:&nbsp; saturacija&nbsp; vode&nbsp; kiseonikom,ukupni&nbsp; organski&nbsp; ugljenik,&nbsp; povr&scaron;inski&nbsp; aktivne materije, temperatura, elektroprovodljivost, pH i ukpni&nbsp; alkalitet.&nbsp; Hidromorfolo&scaron;ki&nbsp; parametri&nbsp; su objasnili&nbsp; 23.48&nbsp; %&nbsp; varijabilnosti&nbsp; u&nbsp; strukturi makrofitske&nbsp; vegetacije.&nbsp; Kao&nbsp; najsignifikantnije varijable,&nbsp; izdvojile&nbsp; su&nbsp; se:&nbsp; struktura&nbsp; vegetacije&nbsp; u priobalnoj&nbsp; zoni,&nbsp; diverzitet&nbsp; prirodnih&nbsp; tipova stani&scaron;ta&nbsp; priobalne&nbsp; zone,&nbsp; prirodnost&nbsp; obale, diverzitet&nbsp; prirodnog&nbsp; supstrata&nbsp; litorala,masimalna dubina &scaron;ljunkara, povr&scaron;ina &scaron;ljunkara,indeks relativne dubine, udaljenost &scaron;ljunkara od glavnog&nbsp; reĉnog&nbsp; toka&nbsp; i&nbsp; starost&nbsp; &scaron;ljunkara. Hidrolo&scaron;ki&nbsp; parametri&nbsp; su&nbsp; objasnili&nbsp; 8.38&nbsp; % varijabilnosti u strukturi&nbsp; makrofitske vegetacije. Kao&nbsp; najsignifikantnije&nbsp; varijable,&nbsp; izdvojile&nbsp; su&nbsp; se broj plavnih talasa u vegetacionoj sezoni tokom godine u kojoj je vr&scaron;eno uzorkovanje vegetacije i broj plavnih talasa u prolećnom periodu za sve četiri&nbsp; godine.&nbsp; Ovi&nbsp; rezultati&nbsp; potvrđuju&nbsp; direktni destruktivni&nbsp; uticaj&nbsp; plavnih&nbsp; talasa&nbsp; na&nbsp; vodenu vegetaciju u vegetacionoj sezoni, kao i indirektni uticaj prolećnih poplava, usled uticaja na trofički status&nbsp; vode.&nbsp; Sumarno,&nbsp; &scaron;ljunkare&nbsp; u&nbsp; plavnom području&nbsp; reke&nbsp; Drine&nbsp; predstavljaju&nbsp; optimalno stani&scaron;te&nbsp; za&nbsp; razvoj&nbsp; retke&nbsp; i&nbsp; ugroţene&nbsp; makrofitske flore.&nbsp; Od&nbsp; ukupnog&nbsp; broja&nbsp; zabeleženih&nbsp; vrsta makrofita,&nbsp; 30&nbsp; %&nbsp; se&nbsp; kategori&scaron;e&nbsp; kao&nbsp; za&scaron;tićeno&nbsp; ili ugroženo na nacionalnom nivou.&nbsp; Značajno veće vrednosti&nbsp; konzervacionih&nbsp; indeksa&nbsp; ustanovljene su&nbsp; za&nbsp; &scaron;ljunkare&nbsp; u&nbsp; poređenju&nbsp; sa&nbsp; prirodnim fluvijalnim jezerima, &scaron;to ukazuje na njihov visok ekolo&scaron;ki&nbsp; potencijal.&nbsp; Ustanovljen&nbsp; je&nbsp; visok diverzitet&nbsp; prioritetnih&nbsp; tipova&nbsp; akvatiĉnih&nbsp; stani&scaron;ta prema&nbsp; Pravilniku&nbsp; o&nbsp; kriterijumima&nbsp; za&nbsp; izdvajanje tipova&nbsp; stani&scaron;ta,&nbsp; o&nbsp; tipovima&nbsp; stani&scaron;ta,&nbsp; osetljivim, ugroženim,&nbsp; retkim&nbsp; i&nbsp; za&nbsp; za&scaron;titu&nbsp; prioritetnim tipovima&nbsp; stani&scaron;ta&nbsp; i&nbsp; o&nbsp; merama&nbsp; za&scaron;tite&nbsp; za&nbsp; njihovo očuvanje, Aneksu I, Direktive Evropske unije oza&scaron;titi&nbsp; prirodnih&nbsp; stani&scaron;ta&nbsp; i&nbsp; divlje&nbsp; flore&nbsp; i&nbsp; faune (Natura&nbsp; 2000),&nbsp; Rezoluciji&nbsp; br.&nbsp; 4&nbsp; Konvencije&nbsp; o očuvanju&nbsp; evropske&nbsp; divlje&nbsp; flore&nbsp; i&nbsp; faune&nbsp; i prirodnih&nbsp; stani&scaron;ta (EMERALD)&nbsp; i&nbsp; Evropskoj crvenoj&nbsp; listi&nbsp; stani&scaron;ta.&nbsp; Vrednosti&nbsp; izdvojenih atributa&nbsp; &scaron;ljunkara&nbsp; mogli&nbsp; bi&nbsp; se&nbsp; iskoristiti&nbsp; u procesu&nbsp; ranog&nbsp; planiranja&nbsp; i&nbsp; projektovanja eksploatacionih&nbsp; polja&nbsp; u&nbsp; plavnom&nbsp; području&nbsp; reke Drine&nbsp; i&nbsp; na&nbsp; drugim,&nbsp; sličnim&nbsp; lokalitetima. Generalna preporuka je da se dva tipa &scaron;ljunkara kreiraju&nbsp; u&nbsp; okviru&nbsp; jednog&nbsp; eksploatacionog&nbsp; polja. Prvi&nbsp; tip,&nbsp; odnosno&nbsp; &scaron;ljunkare&nbsp; koje&nbsp; bi&nbsp; podržavale pionirsku&nbsp; vegetaciju&nbsp; pr&scaron;ljenčica&nbsp; trebale&nbsp; bi&nbsp; da budu locirane na razdaljini do 100 m od glavnog rečnog&nbsp; toka,&nbsp; povr&scaron;ine&nbsp; do&nbsp; 1000&nbsp; m 2 i&nbsp; da&nbsp; imaju vrednost indeksa relativne dubine &gt; 5 %. Drugi tip&nbsp; &scaron;ljunkara&nbsp; koje&nbsp; bi&nbsp; podržavale&nbsp; vegetaciju karakterističnu&nbsp; za&nbsp; nizijska&nbsp; fluvijalna&nbsp; jezera trebale bi da budu locirane na razdaljani od oko 300 m od glavnog rečnog toka, dubine 3&nbsp; &ndash;&nbsp; 4&nbsp; m (najmanje 2 m), povr&scaron;ine između 10000 i 20000 m ²(najmanje&nbsp; 4000&nbsp; m² ),&nbsp; različitih&nbsp; vrednosti indeksa relativne dubine, ali&nbsp; ne preko 5 %. Sva eksploataciona&nbsp; polja&nbsp; bi&nbsp; trebalo&nbsp; isplanirati&nbsp; i isprojektovati&nbsp; kako&nbsp; bi&nbsp; se&nbsp; minimizirao&nbsp; uticaj&nbsp; na priobalnu&nbsp; i&nbsp; obalnu&nbsp; zonu.&nbsp; Pridržavanjem&nbsp; datih smernica&nbsp; povećao&nbsp; bi&nbsp; se&nbsp; diverzitet&nbsp; i&nbsp; kvalitet stani&scaron;ta,&nbsp; kao&nbsp; i&nbsp; konzervacioni&nbsp; potencijal &scaron;ljunkara. Kreiranjem &scaron;ljunkara na naĉin kao &scaron;to je predloženo u ovoj disertaciji omogućila bi se spontana&nbsp; rekultivacija&nbsp; eksploatacionih polja,odnosno&nbsp; remedijacija&nbsp; u&nbsp; cilju&nbsp; pobolj&scaron;anja kvaliteta vode i renaturalizacija stani&scaron;ta, čime bi se&nbsp; znatno&nbsp; smanjili,&nbsp; ili&nbsp; u&nbsp; potpunosti&nbsp; eliminisali, tro&scaron;kovi tehničke rekultivacije terena.</p> / <p>Gravel pit lakes in the river floodplains represent a kind of ecological paradox. Gravel exploitation was recognised&nbsp; as&nbsp; important&nbsp; factor&nbsp; significantlyaffecting&nbsp; aquatic&nbsp; habitats&nbsp; and&nbsp; biodiversity.&nbsp; On the&nbsp; other&nbsp; hand,&nbsp; gravel&nbsp; pit&nbsp; lakes&nbsp; are&nbsp; valuable biodiversity&nbsp; refugiums,&nbsp; potentially&nbsp; supporting rarae&nbsp; species&nbsp; and&nbsp; habitats.&nbsp; The&nbsp; aim&nbsp; of&nbsp; this dissertation&nbsp; was&nbsp; to&nbsp; determine&nbsp; the&nbsp; most significant&nbsp; and&nbsp; relevant&nbsp; hydromorphological parameters&nbsp; in&nbsp; structuring&nbsp; macrophyte assemblages&nbsp; in gravel pit&nbsp; lakes along the Drina River floodplain and to determine the correlation between&nbsp; selected&nbsp; parameters&nbsp; and&nbsp; macrophyte quantitative&nbsp; indices.&nbsp; The&nbsp; research&nbsp; was&nbsp; carried out at the 18 gravel pit lakes (60 survey sectors) in&nbsp; Crna&nbsp; Bara,&nbsp; Badovinci&nbsp; and&nbsp; Lipnicki&nbsp; Sor&nbsp; and four natural fluvial lakes (13 survey sectors), in the&nbsp; Drina&nbsp; River&nbsp; floodplain&nbsp; during&nbsp; the&nbsp; summer months&nbsp; of&nbsp; 2015,&nbsp; 2016,&nbsp; 2017&nbsp; and&nbsp; 2018.Macrophyte&nbsp; vegetation&nbsp; was&nbsp; recorded&nbsp; in&nbsp; all&nbsp; 18 gravel pit lakes, in total supporting 31 taxa.&nbsp; The most&nbsp; abundant&nbsp; species,&nbsp; with&nbsp; highest&nbsp; tot al&nbsp; cover value&nbsp; were <em>Potamogeton</em>&nbsp; <em>nodosus,Ceratophyllum&nbsp;</em> demersum&nbsp; subsp.&nbsp; demersum,<em> Myriophyllum&nbsp; spicatum,&nbsp;</em> <em>Najas&nbsp; marina&nbsp;</em> and<em> Chara&nbsp; globularis</em>.&nbsp; Fluvial&nbsp; lakes&nbsp; supported&nbsp; 13 macrophyte&nbsp; taxa&nbsp; with&nbsp;<em> Vallisneria&nbsp; spiralis, Elodea&nbsp; canadensis,&nbsp; Callitriche&nbsp; palustris,Potamogeton&nbsp;</em> <em>natans&nbsp;</em> and&nbsp;<em> Nuphar&nbsp; lutea</em>&nbsp; as constant and dominant&nbsp; species.&nbsp; The values of all macrophyte&nbsp; quantitative&nbsp; indices&nbsp; found&nbsp; to&nbsp; be significantly&nbsp; higher&nbsp; in&nbsp; the&nbsp; gravel&nbsp; pit&nbsp; lakes compared&nbsp; to&nbsp; the&nbsp; fluvial&nbsp; ones.&nbsp; The&nbsp; cluster analysis&nbsp; revealed&nbsp; 14&nbsp; aquatic&nbsp; vegetation&nbsp; groups (VG).&nbsp; At&nbsp; 16&nbsp; out&nbsp; of&nbsp; 18&nbsp; gravel&nbsp; pit&nbsp; lakes&nbsp; 13 vegetation&nbsp; groups&nbsp; were&nbsp; revealed:&nbsp; VG1<em>Ceratophyllum&nbsp; demersum</em>,&nbsp; VG2&nbsp; <em>Ceratophyllum demersum- Valisneria&nbsp; spiralis</em>,&nbsp; VG3&nbsp;<em> Chara contraria</em>,&nbsp; VG4&nbsp; <em>Chara&nbsp; globularis</em>,&nbsp; VG5&nbsp;<em> Elodea&nbsp; canadensis,</em>&nbsp; VG6&nbsp;<em> Elodea&nbsp; nuttallii,</em>&nbsp; VG7&nbsp; <em>Najas marina,&nbsp;</em> VG8&nbsp;<em> Najas&nbsp; minor,</em>&nbsp; VG9&nbsp; <em>Nitellopsis obtusa</em>,&nbsp; VG10&nbsp; <em>Nuphar&nbsp; lutea,</em>&nbsp; VG11 <em>Potamogeton&nbsp; nodosus</em>,&nbsp; VG12&nbsp; <em>Potamogeton natans,</em> VG13&nbsp;<em> Potamogeton pectinatus</em>), Natural fluvial&nbsp; lakes&nbsp; supported&nbsp; 4&nbsp; vegetation&nbsp; groups: VG5&nbsp; <em>Elodea&nbsp; canadensis</em>,&nbsp; VG10&nbsp; <em>Nuphar&nbsp; lutea</em>, VG12&nbsp; <em>Potamogeton&nbsp; natans&nbsp;</em> and&nbsp; VG14&nbsp;<em> Typha latifolia</em>.&nbsp; All&nbsp; gravel&nbsp; pit&nbsp; lakes&nbsp; can&nbsp; be characterized&nbsp; as&nbsp; water&nbsp; bodies&nbsp; with&nbsp; good&nbsp; to maximal&nbsp; ecological&nbsp; potential,&nbsp; while&nbsp; all&nbsp; the fluvial ones can be characterized as water bidies with&nbsp; poor&nbsp; to&nbsp; moderate&nbsp; ecological&nbsp; status.&nbsp; The values&nbsp; of&nbsp; total&nbsp; suspended&nbsp; supstances,&nbsp; chemical and&nbsp; biological&nbsp; oxygen&nbsp; demand,&nbsp; total&nbsp; organic carbon&nbsp; and&nbsp; nitrates&nbsp; were&nbsp; significantly&nbsp; higher&nbsp; in the natural&nbsp; fluvial&nbsp; lakes compared to the gravel pit&nbsp; ones.&nbsp; Measured&nbsp; level&nbsp; of&nbsp; physico-chemical parameters&nbsp; indicating&nbsp; mesotrophic&nbsp; character&nbsp; of gravel&nbsp; pit&nbsp; lakes&nbsp; in&nbsp; Badovinci&nbsp; and&nbsp; mesoeutrophic&nbsp; in&nbsp; Lipnicki&nbsp; Sor,&nbsp; while&nbsp; all&nbsp; the&nbsp; gravel pits&nbsp; in&nbsp; Crna&nbsp; Bara&nbsp; could&nbsp; be&nbsp; characterized&nbsp; as eutrophic.&nbsp; Similar&nbsp; range&nbsp; values&nbsp; were&nbsp; calculated for&nbsp; LHQA&nbsp; for&nbsp; gravel&nbsp; pit&nbsp; and&nbsp; fluvial&nbsp; lakes&nbsp; (36 &ndash; 49). However, natural lakes&nbsp; showed significantly higher&nbsp; values&nbsp; for&nbsp; LHMS&nbsp; score.&nbsp; The&nbsp; above mentioned,&nbsp; indicates&nbsp; higher&nbsp; anthropogenic pressures&nbsp; on&nbsp; natural&nbsp; fluvial&nbsp; lakes&nbsp; compared&nbsp; to gravel&nbsp; pit&nbsp; ones.&nbsp; Physico-chemical&nbsp; and hydromorphological&nbsp; parameters&nbsp; together explained&nbsp; about&nbsp; 57&nbsp; %&nbsp; of&nbsp; the&nbsp; total&nbsp; variance&nbsp; of macrophyte&nbsp; assemblages&nbsp; with&nbsp; 16.57&nbsp; %&nbsp; of&nbsp; the shared effect. After accounting for the effects of physico-chemical&nbsp; parameters&nbsp; (17.02&nbsp; %), hydromorphological&nbsp; variables&nbsp; explained&nbsp; around 23 % of the total variance.&nbsp; The most significant water&nbsp; quality variables&nbsp; were: oxygen saturation, total&nbsp; organic&nbsp; carbon,&nbsp; surfactants, ,electroconductivity,&nbsp; pH&nbsp; and total alkalinity. The The most significant hydromorphology&nbsp; variables for&nbsp; structuring&nbsp; macrophyte&nbsp; assemblages&nbsp; were: riparian&nbsp; vegetation&nbsp; structural&nbsp; complexity, diversity&nbsp; of&nbsp; natural&nbsp; landcover&nbsp; types&nbsp; in&nbsp; riparianzone, shore structural habitat&nbsp; diversity, diversity of&nbsp; natural littoral zone, maximal&nbsp; lake depth, lake surface&nbsp; area,&nbsp; relative&nbsp; depth&nbsp; ratio,&nbsp; lake&nbsp; distance from&nbsp; r iver&nbsp; main&nbsp; channel&nbsp; and&nbsp; lake&nbsp;&nbsp; age.Hydrologycal parameters were explained 8.38 % of&nbsp; variance&nbsp; in&nbsp; structuring&nbsp; macrophyte assemblages.&nbsp; The&nbsp; most&nbsp; significant&nbsp; hydrology variables&nbsp; were&nbsp; the&nbsp; number&nbsp; of&nbsp; floods&nbsp; in vegetation&nbsp; season&nbsp; in&nbsp; first&nbsp; year&nbsp; when&nbsp; vegetation was sampled, and the number of spring floods in all four research years. These results&nbsp;&nbsp; confirm the direct destructive influence of summer floods&nbsp; on aquatic vegetation, as&nbsp;&nbsp; well as the indirect impact of&nbsp; spring&nbsp; floods,&nbsp; due&nbsp; to&nbsp; the&nbsp; impact&nbsp; on&nbsp; trophic status&nbsp; of&nbsp; water.&nbsp; Gravel&nbsp; pit&nbsp; lakes&nbsp; in&nbsp; te&nbsp; Drina River floodplain&nbsp; represent an optimal habitat&nbsp;&nbsp; for rare&nbsp; and&nbsp; threatened&nbsp; macrophyte&nbsp; flora.&nbsp; Of&nbsp; the total&nbsp; macrophyte&nbsp; species&nbsp; recorded,&nbsp; 30&nbsp; %&nbsp; were categorized&nbsp; as&nbsp; protected&nbsp; or threatened.&nbsp; At&nbsp; least one&nbsp; strictly&nbsp; protected,&nbsp; protected&nbsp; or&nbsp; threatened species&nbsp; was&nbsp; recorded&nbsp; in&nbsp; each&nbsp; gravel&nbsp; pit&nbsp; lake. Significantly&nbsp; higher&nbsp; values&nbsp; of&nbsp; conservation indices&nbsp; (C&nbsp; and&nbsp; Csp&nbsp; score)&nbsp; found&nbsp; to&nbsp; be significantly&nbsp; higher&nbsp; in&nbsp; the&nbsp; gravel&nbsp; pit&nbsp; lakes compared&nbsp; to&nbsp; the&nbsp; fluvial&nbsp; ones.&nbsp; High&nbsp; habitat diversity&nbsp; and&nbsp; conservation&nbsp; value&nbsp; of&nbsp; the&nbsp; sites have&nbsp; been&nbsp; recorded&nbsp; according&nbsp; to&nbsp; the&nbsp; National Rulebook,&nbsp; Annex&nbsp; I&nbsp; of&nbsp; Habitats&nbsp; Directive (NATURA 2000), Resolution&nbsp; no. 4 of the Bern Convention (EMERALD) and the European Red List&nbsp; of&nbsp; Habitats.&nbsp; Values&nbsp; of&nbsp; selected&nbsp; lake attributes can be used for early-design phases of future&nbsp; gravel&nbsp; extraction&nbsp; in&nbsp; the&nbsp; Drina&nbsp; River floodplain&nbsp; area,&nbsp; and&nbsp; in&nbsp; other&nbsp; similar&nbsp; sites. Therefore,&nbsp; general&nbsp; recommendations&nbsp; are&nbsp; that two gravel pit types should be excavated within the&nbsp; single&nbsp; extraction&nbsp; area&nbsp; in&nbsp; order&nbsp; to&nbsp; support pioneering charophyte vegetation and vegetation of typical eutrophic lowland floodplain lakes as well.&nbsp; The&nbsp; first&nbsp; hydromorphological&nbsp; lake&nbsp; type, suitable&nbsp; for&nbsp; stonewort&nbsp; species,&nbsp; should&nbsp; be excavated up to 100 m from river main channel, saving&nbsp; a&nbsp; surface&nbsp; area&nbsp; up&nbsp; to&nbsp; 1000&nbsp; m 2 and&nbsp; a relative depth ratio &gt; 5 %. The second gravel pit type&nbsp; should&nbsp; be&nbsp; located&nbsp; about&nbsp; 300&nbsp; m&nbsp; from&nbsp; river main channel, with preferable maximal depth inrange&nbsp; 3&ndash;4&nbsp; m&nbsp; (at&nbsp; least&nbsp; 2&nbsp; m&nbsp; depth),&nbsp; and&nbsp; a&nbsp; lake surface area between 10000 m 2 and 20000 m 2 (at least&nbsp; 4000&nbsp; m 2 ).&nbsp; Relative&nbsp; depth&nbsp; ratio&nbsp; may&nbsp; vary, but should be less than 5 %. Generally, all sites should&nbsp; be&nbsp; designed&nbsp; with&nbsp; the&nbsp; minimal&nbsp; impact to the&nbsp; riparian&nbsp; and&nbsp; shore&nbsp; zones.&nbsp; These&nbsp; proposed measures&nbsp; would&nbsp; considerably&nbsp; increase&nbsp; lake habitat diversity and their conservation potential. Creating&nbsp; gravel&nbsp; pit&nbsp; lakes&nbsp; as&nbsp; proposed&nbsp; in&nbsp; this dissertation&nbsp;&nbsp;&nbsp; would&nbsp; allow&nbsp; spontaneous recultivation&nbsp; of&nbsp; exploitation&nbsp; fields,&nbsp; remediation in&nbsp; order&nbsp; to&nbsp; improve&nbsp; water&nbsp; quality&nbsp; and renaturalization&nbsp; of&nbsp; habitats,&nbsp; which&nbsp; will significantly reduce, or completely eliminate, the costs of terrain technical recultivation.</p>

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-18T19:21:27Z No. of bitstreams: 1 2010 - Luiza D'Oliveira Sant'Ana.pdf: 2146437 bytes, checksum: 5d8a20463f01fc73a88b5b0c2e56d28c (MD5) / Made available in DSpace on 2017-05-18T19:21:27Z (GMT). No. of bitstreams: 1 2010 - Luiza D'Oliveira Sant'Ana.pdf: 2146437 bytes, checksum: 5d8a20463f01fc73a88b5b0c2e56d28c (MD5) Previous issue date: 2010-07-05 / This work aimed at the evaluation of the total phenolic and flavonoid contents of sixty Apis mellifera honey samples, commercialized in different regions of the state of Rio de Janeiro (Metropolis, Southern, Central, Northwestern and Seaside). For each sample, the total phenolic and the total flavonoid contents were analyzed by the Folin-Denis and aluminum chloride (AlCl3) methods, respectively. Also, the antioxidant activity (%AA and EC50) for all the honey samples was evaluated, as well as for some phenolic acids and flavonoids, used as standards. The determination of the antioxidant capacity was carried out by the method of trapping the 2,2-diphenyl-1-picryl-hydrazyl organic radical (DPPH) by antioxidant compounds, resulting in an absorbance decrease at 520 nm. The analyzed honeys were classified as monofloral or heterofloral. For the monofloral samples, it was observed that Anadenanthera honeys showed the highest average value for total phenolics (112,60 mgEAG/100g) and also for the antioxidant capacity (59,93 (AA)mgEQC/500g or 18,92 mg/mL if expressed as EC50), while the Eucalyptus honeys showed the highest mean for total flavonoids (9,17 mgEQC/100g). With these results, a comparative study was carried out with diverse honey samples of different regions, in terms of total phenolic and flavonoid content, antioxidant capacity and some physico-chemistry parameters (HMF, acidity, pH, humidity, water activity and color intensity). Comparison of these results allowed to observe that darker honeys showed both higher antioxidant activity and the highest values for total phenolic and flavonoid contents, as well as for all the other physico-chemical parameters. / O objetivo deste trabalho foi avaliar os teores de fen?licos e flavon?ides totais de sessenta amostras de mel de Apis mellifera comercializadas em diferentes mesorregi?es do estado do Rio do Janeiro (Metropolitana, Sul, Centro, Noroeste Fluminense e Baixada Litor?nea). Para cada amostra foram analisados os totais de fen?licos e flavon?ides, atrav?s dos m?todos de Folin-Denis e cloreto de alum?nio (AlCl3), respectivamente. Foi determinado, tamb?m, o potencial da atividade antioxidante (%AA e CE50) para todas as amostras de mel, bem como para alguns padr?es de ?cidos fen?licos e flavon?ides. O m?todo utilizado para determinar a capacidade antioxidante foi o de captura do radical org?nico 2,2-difenil-1-picril-hidrazil (DPPH) por antioxidantes, produzindo um decr?scimo da absorb?ncia a 520 nm. Os m?is avaliados foram classificados de monoflorais e heteroflorais. Para os monoflorais foi observado que os m?is de Anadenanthera apresentaram a maior m?dia em totais de fen?licos (112,60 mgEAG/100g), e tamb?m a maior atividade antioxidante (59,93 (AA)mgEQC/500g ou 18,92 mg/mL em termos de CE50), enquanto os m?is de Eucalyptus apresentaram, em m?dia, o maior conte?do de flavon?ides totais (9,17 mgEEC/100g). A partir dos resultados obtidos foi feito um estudo comparativo entre as v?rias amostras de m?is de diferentes regi?es quanto ao teor de fen?licos e flavon?ides totais, a atividade antioxidante e algumas propriedades f?sico-qu?micas (HMF, acidez, pH, umidade, atividade de ?gua e intensidade de colora??o). Ao comparar esses resultados, foi poss?vel observar que os m?is mais escuros apresentaram uma maior atividade antioxidante e uma maior quantidade de fen?licos e flavon?ides totais, bem como valores mais altos para todos os outros par?metros f?sico-qu?micos comparados.

Στα πλαίσια αυτής της διδακτορικής εργασίας μελετήθηκαν οι εμπορικά σημαντικότερες περιοχές καλλιέργειας και συγκομιδής οστρακοειδών του Ελλαδικού χώρου. Κατά τη διάρκεια περιόδου 18 μηνών πραγματοποιήθηκε μηνιαία συλλογή δειγμάτων στρειδιών (Οstrea edulis) και μυδιών (Mytilus galloprovincialis), τα οποία συλλέχθηκαν από έξι (6) διαφορετικά σημεία του Ελλαδικού χώρου και αναλύθηκαν για τους εντεροϊούς (EV), τους αδενοϊούς (Adv), τον ιό της ηπατίτιδας Α (HAV), τους ιούς Noro I και II (NLVI και NLVII), για το βακτήριο Ε. coli, καθώς και για σωματικούς κολιφάγους, τους F-sperific RNA βακτηριοφάγους και τους βακτηριοφάγους του Β. fragilis. Επιπλέον αναπτύχθηκαν μέθοδοι τόσο για την ανίχνευση παθογόνων ιών ανθρώπινης προέλευσης στα οστρακοειδή, όσο και για την ανίχνευση των "πιθανών δεικτών" αυτών των ιών. Οι μέθοδοι εξετάστηκαν προκειμένου να αξιολογηθεί η απόδοση καλής ποιότητας από όλα τα εργαστήρια μέσω διεργαστηριακών αναλύσεων. Η μέθοδος που εφαρμόστηκε σε αυτή τη μελέτη για την ανίχνευση των ιών στα οστρακοειδή βασίζεται στην εξαγωγή και την ομογενοποίηση του πεπτικού αδένα με χρήση διαλύματος γλυκίνης, pH 10, απομόνωση των νουκλεϊνικών οξέων και ενίσχυση του γονιδιώματος των ιών που αναλύονται. Για την ανίχνευση του βακτηρίου E. coli χρησιμοποιήθηκε η μέθοδος των πολλαπλών σωλήνων, ενώ για την ανίχνευση των βακτηριοφάγων χρησιμοποιήθηκε η μέθοδος καλλιέργειας διπλοστιβάδας. Για το βακτήριο E. coli, σε σύνολο 138 δειγμάτων, 110 δείγματα (ποσοστό 79,7%) βρέθηκαν να ανήκουν στην κατηγορία Α (MPN/100g σάρκας = <20 έως 220), δηλαδή χαρακτηρίζονται σαν δείγματα χαμηλής μόλυνσης, 25 δείγματα (ποσοστό 18,1%) βρέθηκαν να ανήκουν στην κατηγορία Β (MPN/100g σάρκας = 220 έως 3500), οπότε χαρακτηρίζονται σαν δείγματα μεσαίας μόλυνσης, ακατάλληλα προς κατανάλωση χωρίς να προηγηθεί διαδικασία εξυγίανσης, ενώ μόνο 3 δείγματα (ποσοστό 2,2%) βρέθηκαν να ανήκουν στη κατηγορία C (MPN/100g σάρκας =3500 έως >18000), δηλαδή είναι δείγματα υψηλής μόλυνσης. Οι ιοί που εμφανίζονται με μεγαλύτερη συχνότητα στα οστρακοειδή της Ανατολικής Μεσογείου είναι οι αδενοϊοί (34% των δειγμάτων βρέθηκαν θετικά για τους αδενοϊούς) και ακολουθούν οι εντεροϊοί (16,7% των δειγμάτων βρέθηκαν θετικά για τους εντεροϊούς). Αντίθετα, ο ιός της ηπατίτιδας Α (ποσοστό θετικών δειγμάτων = 4,34%), καθώς και οι ιοί Noro I (ποσοστό θετικών δειγμάτων = 2,1%) και Noro II (ποσοστό θετικών δειγμάτων = 1,47%%) εμφανίζονται σε μικρό ποσοστό δειγμάτων. Τέλος, 80 δείγματα (58%) βρέθηκαν θετικά (παρουσία πλακών βακτηριοφάγων) για τους σωματικούς κολιφάγους, με τον αριθμό των πλακών να κυμαίνεται από 71,4 έως 584800 pfp/100g, 52 δείγματα (37,7%) βρέθηκαν θετικά για τους F-specific RNA βακτηριοφάγους (αριθμός των πλακών από 76,2 έως 17051 p100g) και 33 δείγματα (24%) βρέθηκαν θετικά για τους βακτηριοφάγους του Bacteroides fragilis (αριθμός των πλακών από 194.5 έως 5266,25 pfp/100g). Τόσο για το βακτήριο E. coli όσο και για τους βακτηριοφάγους πραγματοποιήθηκαν διεργαστηριακές αναλύσεις προτύπων, οι οποίες οδήγησαν στο συμπέρασμα ότι οι αντίστοιχες μέθοδοι χαρακτηρίζονται ως αξιόπιστες. Η στατιστική ανάλυση έδειξε ότι το βακτήριο E. coli παρουσιάζει θετική συσχέτιση με τους σωματικούς κολιφάγους, αλλά δεν δείχνει στατιστικά σημαντική συσχέτιση ούτε με τους F-specific RNA βακτηριοφάγους, ούτε με κανέναν από τους ιούς εντερικής προέλευσης. Επίσης, θετική συσχέτιση παρουσίασαν οι αδενοϊοί με τους εντεροϊούς, καθώς και οι σωματικοί κολιφάγοι με τους βακτηριοφάγους του B. fragilis. Η μοναδική συσχέτιση μεταξύ ιών εντερικής προέλευσης και βακτηριοφάγων βρέθηκε για τους αδενοϊούς και τους βακτηριοφάγους του B. fragilis. Εάν αυτό επιβεβαιωθεί σε περαιτέρω μελέτες, τότε η συγκεκριμένη κατηγορία βακτηριοφάγων θα μπορούσε να αποτελέσει έναν καλό δείκτη πρόβλεψης της παρουσίας αδενοϊών σε δείγματα οστρακοειδών. Επιπλέον μελετήθηκε η σχέση που μπορεί να υπάρχει μεταξύ των φυσικοχημικών παραμέτρων και των μικροοργανισμών που εξετάστηκαν. Η επεξεργασία αυτή οδήγησε στο συμπέρασμα ότι το βακτήριο E. coli ανιχνεύεται σε μεγαλύτερα ποσά όταν το διαλυμένο οξυγόνο και η περιεκτικότητα σε άλας του ύδατος είναι αυξημένα. Αντίθετα, αύξηση της θερμοκρασίας οδηγεί σε μείωση της ανίχνευσης του βακτηρίου. Επίσης, η περιεκτικότητα σε άλας φαίνεται να επηρεάζει θετικά και τον ιό της ηπατίτιδας Α, αν και ο μικρός αριθμός θετικών δειγμάτων γι’αυτόν τον ιό δεν μπορεί να επιτρέψει την εξαγωγή ασφαλών συμπερασμάτων. Το pH και το διαλυμένο οξυγόνο των υδάτων οδηγεί σε αύξηση της ανίχνευσης των βακτηριοφάγων του B. fragilis, χωρίς όμως να μπορούμε να ισχυριστούμε ότι κάτι τέτοιο ισχύει, λόγω του μικρού αριθμού θετικών δειγμάτων γι’αυτούς τους βακτηριοφάγους. Τέλος, η αύξηση της θερμοκρασίας των υδάτων φαίνεται να οδηγεί και σε αύξηση της παρουσίας των F-specific RNA βακτηριοφάγων, και το ίδιο παρατηρήθηκε και με την αύξηση του διαλυμένου οξυγόνου στο νερό. Η παρούσα μελέτη αποτελεί την πρώτη διεξοδική έρευνα για την ιογενή κοπρανώδη μόλυνση τον οστρακοειδών στην Ελλάδα. Επιπλέον, αντιπροσωπεύει την πρώτη μελέτη σχετικά με τη αποτελεσματικότητα των οργανισμών - δεικτών ιϊκής μόλυνσης, καθώς και για τη συσχέτιση της μικροβιολογικής επιβάρυνσης των οστρακοειδών με τις φυσικοχημικές παραμέτρους του περιβάλλοντος ύδατος. Η μελέτη κατάλληλων δεικτών που σχετίζονται με την παρουσία εντερικών ιών στα οστρακοειδή οδήγησε σε χρήσιμα συμπεράσματα για τη χρήση της ανίχνευσης των βακτηριοφάγων ως δεικτών ιϊκής μόλυνσης. Εντούτοις, απαιτείται περαιτέρω μελέτη προκειμένου να προσδιοριστεί και η χρήση των βακτηριοφάγων ως δεικτών που θα μαρτυρούν την προέλευση (ανθρώπινη ή ζωική) των εντερικών ιών που ανιχνεύονται στα οστρακοειδή. / In this doctorate investigation, important shellfish growing areas of Greece have been defined and studied. Oysters (Ostrea edulis) and mussels (Mytilus galloprovincialis) were obtained on a monthly basis over an 18 month sampling period. These samples were collected by six (6) different points of Greece and were analyzed for enteroviruses (EV), adenoviruses (Adv), virus of hepatitis A (HAV), Noro viruses I and II (NLVI and NLVII ), bacterium E. coli, as well as for somatic coliphages, F-sperific RNA bacteriophages and bacteriophages of B. fragilis. Moreover, methods were developed for the detection of pathogenic viruses of human origin in the shellfish, as well as for the detection of potential "viral indicators". The methods were examined in order to validate the good quality performance from all the laboratories via interlaboratory analyses. The method that used in this study for the detection of human enteric viruses in the shellfish is based on the export and homogenisation of digestive gland with glycine buffer at pH 10, viral nucleic acid extraction and amplification of the genomes of the analysed human viruses. The procedure applied for detection of E. coli consists on a five tube, three dilution most probable number (MPN) method, while the method for the detection of bacteriophages was the double-agar-layer method. For E. coli analysis, in a total number of 138 samples, 110 samples (79,7%) were found to belong in category A (MPN / 100 g of flesh = < 20 until 220), that it means these samples are characterized as samples of low pollution, 25 samples (18,1%) were found to belong in category B (MPN / 100 g of flesh = 220 until 3500), therefore are characterized as samples of intermediate pollution, inadequate to consumption without precedes process of cleansing, while only 3 samples (2,2%) were found to belong in category C (MPN / 100 g of flesh = 3500 until > 18000), that it means they are samples of high pollution. The viruses that are presented with higher frequency in the shellfish of Eastern Mediterranean are the adenoviruses (34% of samples were found positive for adenoviruses) and follow enteroviruses (16,7% samples they were found positive for enteroviruses). On the contrary, virus of hepatitis A (percentage of positive samples = 4,34%), as well as the Noro I viruses (percentage of positive samples = 2,1%) and Noro II viruses (percentage of positive samples = 1,47%%) are presented in small number of samples. Finally, 80 samples (58%) were found positive (presence of plaques of bacteriophages) for somatic coliphages, with the number of plaques between 71,4 and 584800 pfp / 100 g, 52 samples (37,7%) were found positive for F - specific RNA bacteriophages (number of plaques from 76,2 to 17051 pfp/ 100 g) and 33 samples (24 %) were found positive for the bacteriophages of B. fragilis (number of plaques from 194,5 to 5266,25 pfp / 100 g). Interlaboratory studies involved the testing of reference materials of E. coli and bacteriophages were used as part of the good quality performance assessment program to be applied all over the study, and led to the conclusion that the corresponding methods are characterized by good reliability. According to the statistical analysis of the results, the presence of E. coli seems to be significantly related with the presence of somatic coliphages. However, E. coli do not present significant statistical relation neither with F - specific RNA bacteriophages, nor with all of the viruses of intestinal origin. Also, adenoviruses were significantly related with enteroviruses, as well as somatic coliphages with the bacteriophages of B. fragilis. The unique significant relation between viruses of intestinal origin and bacteriophages was found for the adenoviruses and bacteriophages of B. fragilis. If this is confirmed in further studies, then this category of bacteriophages could constitute a good indicator of forecast of presence adenoviruses in samples of shellfish. Moreover, we studied the relation that can exist between the physic-chemical parameters and the micro-organisms that were examined. This analysis led to the conclusion that E. coli is detected in higher levels when the dissolved oxygen and the salinity of water are increased. On the contrary, increase of temperature leads to reduction of detection of E. coli. Also, the salinity appears to influence positively also virus of hepatitis A, even if the small number of positive samples of this virus cannot allow the export of sure conclusions. The pH and the dissolved oxygen of waters lead to increase of detection of bacteriophages of B. fragilis, but the small number of positive samples for these bacteriophages can’t give safe conclusions. Finally, the increase of temperature of waters appears to lead also to increase of presence of F - specific RNA of bacteriophages, and the same was also observed with the increase of dissolved oxygen in water. This study constitutes the first extensive research for the fecal viral pollution of shellfish in Greece. Moreover, it represents the first study with regard to the effectiveness viral indicators, as well as for the correlation of microbiological parameters of shellfish with the physical-chemical parameters of water. The study of suitable indicators that are related with the presence of enteric viruses in the shellfish led to useful conclusions on the use of detection of bacteriophages as indicators of viral pollution. Nevertheless, further study is required in order to determine also the use of bacteriophages as indicators that will testify the origin (human or animal) of the enteric viruses that are detected in the shellfish.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-03-13T17:29:42Z No. of bitstreams: 1 andressasilvinoferreiraassis.pdf: 3364177 bytes, checksum: ff9c20080e11e36f28daf805d56ecc7e (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-03-13T19:33:58Z (GMT) No. of bitstreams: 1 andressasilvinoferreiraassis.pdf: 3364177 bytes, checksum: ff9c20080e11e36f28daf805d56ecc7e (MD5) / Made available in DSpace on 2017-03-13T19:33:58Z (GMT). No. of bitstreams: 1 andressasilvinoferreiraassis.pdf: 3364177 bytes, checksum: ff9c20080e11e36f28daf805d56ecc7e (MD5) Previous issue date: 2016-12-19 / O tratamento de esgoto pode ser insuficiente para a completa eliminação de vírus entéricos, tais como adenovírus humanos (HAdV) e rotavírus do grupo A (RVA). Portanto, o retorno do lodo e do efluente tratado ao ambiente pode representar riscos à saúde pública. Este estudo foi conduzido para otimizar um protocolo de floculação orgânica para recuperação viral a partir de lodo de esgoto e efluente tratado, bem como realizar um monitoramento de HAdV e RVA na estação de tratamento de esgoto (ETE) de Juiz de Fora, MG. Nos estudos de otimização, foram propostas adaptações no protocolo de floculação com leite desnatado para lodo e efluente tratado, com modificações no tempo de agitação da amostra, na concentração final de leite desnatado e/ou na etapa de centrifugação. No estudo de monitoramento, esgoto bruto (P1), esgoto primário (P2), lodo (P3) e efluente tratado (P4) foram coletados bimensalmente em 2014 (durante as épocas seca e chuvosa), totalizando 96 amostras (simples e compostas). As cargas virais foram determinadas por PCR quantitativo e o bacteriófago PP7 foi usado como controle interno. Amostras de HAdV e RVA foram submetidas ao sequenciamento e a viabilidade das partículas de HAdV foi avaliada em amostras de P4. Os parâmetros físico-químicos e a contagem de coliformes termotolerantes (CT) foram determinados em cada ponto. Nos estudos de otimização, foram selecionadas duas condições que apresentaram as maiores taxas de recuperação viral no lodo (menor tempo de agitação e maior concentração de leite desnatado) e no efluente tratado (sem primeira etapa de centrifugação e com maior concentração de leite desnatado). Ambas provaram ser ferramentas úteis para pesquisa viral em amostras de campo, inclusive para a pesquisa de vírus gigantes. No monitoramento, o HAdV foi detectado em 85,4% (82/96) dos concentrados, com cargas virais variando de 3,27 x 102 a 2,42 x 106 cópias do genoma por mililitro (cg/mL), ao longo do ano. A presença de RVA foi observada em 52,1% (50/96) dos concentrados (1,38 x 103 a 3,65 x 105 cg/mL), com maior detecção na época seca. A carga viral não foi influenciada pelo tipo de amostra, sendo detectada tanto em amostras simples, quanto em amostras compostas. Todas as amostras de HAdV sequenciadas pertenciam à espécie F tipo 41 e as amostras de RVA pertenciam ao genótipo I1. O tratamento de esgoto reduziu a quantidade de matéria orgânica e sólidos, bem como a contagem de CT e as cargas virais. No entanto, a presença de HAdV e RVA foi observada mesmo após o tratamento, inclusive em amostras de efluente tratado consideradas adequadas pelas legislações atuais, com detecção de partículas infecciosas de HAdV. Foram observadas correlações positivas entre a carga viral e a demanda bioquímica de oxigênio, os sólidos sedimentáveis e sólidos suspensos totais. Os dois protocolos otimizados neste estudo podem ser facilmente adequados para uso em laboratório de rotina, podendo impulsionar o monitoramento viral nos subprodutos gerados na ETE. A carga viral detectada na ETE salienta a disseminação ambiental de RVA e HAdV e aponta o potencial do HAdV como um marcador viral de contaminação em ambientes aquáticos. / Sewage treatment may be insufficient for the complete elimination of enteric viruses such as human adenoviruses (HAdV) and group A rotaviruses (RVA). Thus, the return of sewage sludge and treated effluent to the environment poses concerns potential for public health. This study was conducted to optimize an organic flocculation procedure for viral recovery from sludge and treated effluent, and carry out a surveillance of HAdV and RVA in a wastewater treatment plant (WWTP) at Juiz de Fora, MG. In optimization studies, conditions were proposed for sludge and treated effluent with changes in the stirring time, in the final concentration of skimmed-milk and/or in the centrifugation step. In surveillance study, raw sewage (P1), primary sewage (P2), sludge (P3) and treated effluent (P4) were collected bimonthly in 2014 (during the dry and the rainy season), totaling 96 samples (simple and composite). Quantitative PCR determined viral loads and PP7 bacteriophage was used as internal control. HAdV and RVA strains were selected for sequencing, and the HAdV viability was evaluated in P4 samples. Physicochemical parameters and thermotolerant coliforms (TC) counting were determined at each point. After the optimization studies, two conditions were selected: the ones that showed the highest viral recovery rates in sludge (lower stirring time and higher concentration of skimmed-milk) and treated effluent (without the first centrifugation step and with a higher concentration of skimmed-milk). These conditions proved to be a useful tool for viral search in the field samples, including for the research of giant virus. In surveillance study, HAdV was detected in 85.4% (82/96) of the concentrated, with viral loads ranging from 3.27 x 102 to 2.42 x 106 genome copies per milliliter (gc/mL), throughout the year. RVA presence was observed in 52.1% (50/96) of the samples (1.38 x 103 to 3.65 x 105 gc/mL) with detection greater in the dry season. Viral load was not influenced by the type of sample being detected both in single samples, as in composite samples. All the sequenced HAdV strains belonged to species F type 41, and RVA strains belonged to genotype I1. Sewage treatment reduced the content of organic matter and solids, as well as the TC counts and the viral loads. However, the presence of HAdV and RVA was observed after treatment, even in samples considered adequate by current laws with detection of infectious HAdV particles. Positive correlations were observed between viral load and biochemical oxygen demand, sedimented solids and total suspended solids. Two optimized protocols in this study are easily suitable for routine laboratory use and can boost viral monitoring in by-products generated in the WWTP. Viral load detected in WWTP stress the environmental dissemination of HAdV and RVA and addressed the potential of HAdV as a virological marker of contamination in aquatic environments.

This master thesis focuses on an analysis of surface water chemistry, long-term trends and the impact of drought on changes in concentrations of selected parameters in the upper Svatava, Rolava and Načetínský potok basins located in the Ore Mountains. Research on water quality in mountain spring areas is important, the consequences of various changes can be easily observed there. This work analyses the changes in surface water chemistry and discusses their possible causes, especially the impact of peat bogs and dry episodes. The parameters of water temperature, conductivity, pH, BOD5, COD, TOC, concentration of nitrate nitrogen, total phosphorus, phosphates, sodium, potassium, calcium and iron were investigated for the period 1993-2018. The available data are analysed using evaluation methods according to ČSN 75 7221 as well as box plots, Pearson correlation coefficient, PCA analysis and Mann-Kendall test. The greatest anthropogenic influence exhibited in Svatava, almost all concentrations reached their highest values there. In the Rolava and Načetínský potok basins, the influence of peatlands manifested itself by increasing the concentrations of iron, TOC and COD. The results of trends showed an increase both in pH and in surface water temperature related to an increase in air temperature, on the...

Inadequately treated wastewater effluent is harmful to the receiving aquatic environment. Water-borne chemicals and microbial pathogens pose a health risk to anyone living downstream from sewage treatment facilities. This study assessed the effluent from a package plant with a design capacity of 48kℓ/24 hours, servicing 12 household units and a restaurant in Mogale City. Over a 12 month period, fortnightly water samples were collected from ten selected sites including two boreholes, a river and two dams. Standard parameters including physical (pH, EC, temperature, DO and SS), chemical (nutrient concentration) and biological (bacterial counts) were analysed using handheld meters, standard membrane filter techniques and colorimetric methods. One borehole was affected by pathogen and nitrate runoff from an adjacent poultry farm. If regularly monitored, the package plant effectively removed microbes (most samples contained 0 cfu/100mℓ) but above limit COD, ammonia and phosphate was released in the effluent (with maximum values of 322 mg/ℓ, 42.52 mg/ℓ and 7.18 mg/ℓ, respectively). Generally, river and dam water at the site was of good quality. / Environmental Science / M. Sc. (Environmental Science)

A tuberculose (TB) é uma doença causada pelo Mycobacterium tuberculosis. De acordo com estimativas da Organização Mundial da Saúde, a tuberculose é responsável pela morte de ~2 a 3 milhões de pessoas/ano no mundo e nos próximos 15 anos cerca de 1 bilhão de pessoas deverão ser infectadas, e destas, aproximadamente 35 milhões deverão morrer. Apesar de existirem vários medicamentos sendo utilizados no tratamento da doença, constatasse o crescimento no número de casos devido, principalmente, às variedades resistentes do M. tuberculosis. Considerando-se o aparecimento de cepas resistentes em TB, recomenda-se que novos medicamentos e/ou alvos biológicos alternativos devam ser intensivamente pesquisados. A ribonucleotídeo redutase (RNR), por exemplo, é uma proteína de interesse, pois catalisa uma etapa importante e única na síntese de novo dos dNTPs, reduzindo o ribonucleosídeo 5\' -difosfato ao seu correspondente desoxirribonucleosídeo 5\' -difosfato. A RNR é importante na síntese do DNA, e portanto, na divisão das células. Esta enzima importante, que possuí 16% de homologia com a RNR de mamíferos, é um alvo potencial para o desenvolvimento de novos fármacos, com provável aplicação no tratamento do câncer, da malária e do tripanossoma. Sabe-se que diferentes classes de compostos, através de diferentes mecanismos de ação, inibem a RNR, incluindo as &#945;-(N)-heterocíclicas carboxaldeído tiossemicarbazonas, um dos inibidores mais potentes da RNR. Sabe-se que alguns derivados da tiossemicarbazona, inibidoras da RNR de células tumorais, apresentam atividade frente o M. tuberculosis atuando provavelmente através do mesmo mecanismo, envolvendo a inibição da correspondente RNR. Neste contexto, nesta tese de doutorado, foram aplicadas diferentes abordagens de QSAR/QSAR-3D no estudo de 40 derivados da 2-piridino-carboxaldeído tiossemicarbazona, inibidores da RNR de células H.Ep.-2, retirados de literatura selecionada (French & Blanz-Jr. 1974). Estes compostos foram divididos em cinco séries, a saber: séries A, B, C, D, e E contendo, respectivamente, 40, 39, 30, 23 e 22 compostos, na tentativa de tornar estas séries estruturalmente mais homogêneas. Para cada série, foram criados três grupos de treinamento e os respectivos grupos de teste (I, II e III), visando-se avaliar o poder de predição dos modelos gerados através das análises de QSAR/QSAR-3D. Para as análises de QSAR clássico, foram utilizados como variáveis independentes, os descritores mais relevantes gerados através do programa DRAGON e, pré-selecionados por PLS. Considerando-se a ausência de informações sobre a estrutura cristalográfica da enzima RNR do M. tuberculosis, os estudos de QSAR-3D foram iniciados empregando-se metodologias propostas em CoMFA e, em CoMSIA, implementadas no programa SYBYL. Além destas, foi realizada a modelagem por homologia da RNR do M. tuberculosis, utilizando-se o programa WHATIF. Para as abordagens CoMFA e CoMSIA as geometrias otimizadas através do método semi-empírico AM1 foram alinhadas átomo-a-átomo e, através da similaridade dos respectivos campos estéricos e eletrostáticos, utilizando-se o programa SEAL. Nos dois procedimentos a geometria do composto não substituído, um dos mais ativos na série, foi utilizada como molde considerando-se a ausência de informações sobre a conformação bioativa. A modelagem da RNR por homologia foi realizada utilizando-se como molde as estruturas cristalográficas, respectivamente, do C. ammoniagenes (código PDB 1KGN) e da S. typhimurium (código PDB 1R2F), sambas apresentando valores de identidade superior a 65%. Mais recentemente foram publicados os dados cristalográficos para a cadeia beta (subunidade menor) da RNR do M. tuberculosis (código PDB 1UZR). Os modelos CoMFA e CoMSIA gerados apresentaram valores aceitáveis para os coeficientes de correlação de predição, com altos valores para os coeficientes de correlação ajustados e baixos valores para os erros padrões. Os melhores modelos CoMFA e CoMSIA foram obtidos considerando o grupo com substituintes apenas na posição 5 do anel piridínico. Razoáveis coeficientes de correlação de predição para os modelos CoMSIA com altos coeficientes de correlação de ajuste e baixos valores para os erros padrões forma obtidos. Os mapas de contorno gerados em CoMFA e CoMSIA sugerem que grupos aceptores de ligações de hidrogênio próximos ao nitrogênio do anel piridínico deverá aumentar o valor da atividade inibitória. Esta observação está em boa concordância com os dados da literatura, na qual a formação de um complexo entre a TSC e o íon Ferro foi sugerido para a inibição da RNR. Estes estudos deverão permitir um melhor entendimento sobre as características estruturais desta classe de TSC inibidoras da RNR, como agentes antitumorais, em termos dos campos estéricos, eletrostáticos, hidrofóbico, doador e aceptor de ligações de hidrogênio, bem como a contribuição para o desenvolvimento racional de novos inibidores para esta importante enzima. Adicionalmente, dois compostos preparados em nosso laboratório, demonstraram atividade frente o M. tuberculosis, em testes realizados in vivo. / Tuberculosis is an illness caused by Mycobacterium tuberculosis. Data from World Health Organization (WHO) estimates, that about 2-3 millions of human population died by Mycobacterium tuberculosis infection and that during the next 15 years about 1 billion will be infected and 35 million will certainly die. Although, in the clinic it was found several antiTBdrugs, these numbers will increase due several reasons including M. tuberculosis resistant strains. It has been stressed the importance of novel medicines and/or alternative biological targets research projects. It is known that Ribonucleotide reductase (RNR), is an enzyme that catalyses the rate limiting step in the de novo synthesis of dNTPs, reducing the ribonucleoside 5\'-diphosphates to the corresponding deoxyribonuc1eoside 5\' -diphosphates. RNR has a critical role in the DNA synthesis and, hence, cell division. This key enzyme, that shows 16% homology when compared with mammals RNR, is a potential target for drug design of cell growth inhibitors, with potential application in cancer therapy, antimalaria and trypanosome chemotherapy. It is known that different types of compounds or species by means of different mechanism pathways can show RNR inhibition, including &#945;-(N)-heterocyclic carboxaldehydes thiosemicarbazones that are one of the most potent classes of RNR inhibitors. More than that, some of them, that shows activity against M. tuberculosis seems to follow the same mechanism pathways proposed to the thiosemicarbazones tumor cells activity that means, that they probably are RNR inhibitors. In this study, a series of 40 &#945;-(N)-2-formyl-pyridine thiosemicarbazone derivatives tested against RNR of H.ep.-2-cells (human epidermoid carcinoma), taken from selected literature (French & Blanz-Jr. 1974), has quantitatively analyzed by means of several QSAR/3D-QSAR approaches. These compounds were divided into 5 individual subsets, namely A, B, C, D, and E, having 40, 39, 30, 23 e 22 compounds, respectively. This procedure has been done in order to achieve more structurally homogeneous subsets. For each set, three individual training and test sets (I,II and III) have been created in order to evaluate the predictivity power of the generated QSAR/3D-QSAR models. QSAR analysis have been done using descriptors generated by DRAGON program that have been further pre-selected by PLS procedures. Considering that crystallographic data of RNR M. tuberculosis are not available in the literature, 3D-QSAR studies have been done these applying, initially, CoMFA and CoMSIA approaches, implemented in SYBYL. Homology model studies have been performed with WHATIF program CoMFA e CoMSIA approaches used optimized geometry obtained by semi-empirical AM1 methods that have been aligned by two different methods. Rigid alignment, in which the compounds were fitted atom-by-atom onto a template, based on the root mean square fit. The N(l) and C(2) atoms of the pyridine moiety and the heavy atoms of thiosemicarbazone backbone of TSC were used as template structure. (2) Field based, in which the steric and electrostatic fields, generated by the SEAL program were considered in the alignment. In both procedures the unsubstituted 2-formylpyridine thiosemicarbazone in its syn conformation, has been taken as template. Homology RNR models were done using as template crystallographic data of ammoniagenes (1KGN) and S. typhimurium (1R2F) as template, respectively, with identity larger than 65%. More recent1y new crystallographic data have been published for the beta chain (smaller subunity) of RNR do M. tuberculosis (1UZR). CoMFA and CoMSIA generated models showed acceptable predictive correlation coefficients with high fitted correlation coefficients and low standard errors. Betler CoMFA and CoMSIA models have been derived considering a homogeneous subset of TSC substituted only at 5-position in pyridine ring. Reasonable predictive correlation coefficients for CoMSIA models with high fitted correlation coefficients and very low standard errors were obtained. The derived CoMFA and CoMSIA countour maps suggested that a hydrogen bond acceptor near the nitrogen pyridine ring could enhance inhibitory activity value. This observation is in good agreement with literature, in which a complex formation between TSC and iron ion has been suggested, to RNR inhibition. These studies are expected to enhance the understanding of the structural features of this class of TSC-RNR inhibitors as antitumor agents in terms of steric, electrostatic, hydrophobic and hydrogen donor and acceptor fields as well as to contribute to rational design of inhibitors of this key enzyme. Additionally, two compounds that have been prepared by us showed activity against M. tuberculosis using in vivo test system.

A tuberculose (TB) é uma doença causada pelo Mycobacterium tuberculosis. De acordo com estimativas da Organização Mundial da Saúde, a tuberculose é responsável pela morte de ~2 a 3 milhões de pessoas/ano no mundo e nos próximos 15 anos cerca de 1 bilhão de pessoas deverão ser infectadas, e destas, aproximadamente 35 milhões deverão morrer. Apesar de existirem vários medicamentos sendo utilizados no tratamento da doença, constatasse o crescimento no número de casos devido, principalmente, às variedades resistentes do M. tuberculosis. Considerando-se o aparecimento de cepas resistentes em TB, recomenda-se que novos medicamentos e/ou alvos biológicos alternativos devam ser intensivamente pesquisados. A ribonucleotídeo redutase (RNR), por exemplo, é uma proteína de interesse, pois catalisa uma etapa importante e única na síntese de novo dos dNTPs, reduzindo o ribonucleosídeo 5\' -difosfato ao seu correspondente desoxirribonucleosídeo 5\' -difosfato. A RNR é importante na síntese do DNA, e portanto, na divisão das células. Esta enzima importante, que possuí 16% de homologia com a RNR de mamíferos, é um alvo potencial para o desenvolvimento de novos fármacos, com provável aplicação no tratamento do câncer, da malária e do tripanossoma. Sabe-se que diferentes classes de compostos, através de diferentes mecanismos de ação, inibem a RNR, incluindo as &#945;-(N)-heterocíclicas carboxaldeído tiossemicarbazonas, um dos inibidores mais potentes da RNR. Sabe-se que alguns derivados da tiossemicarbazona, inibidoras da RNR de células tumorais, apresentam atividade frente o M. tuberculosis atuando provavelmente através do mesmo mecanismo, envolvendo a inibição da correspondente RNR. Neste contexto, nesta tese de doutorado, foram aplicadas diferentes abordagens de QSAR/QSAR-3D no estudo de 40 derivados da 2-piridino-carboxaldeído tiossemicarbazona, inibidores da RNR de células H.Ep.-2, retirados de literatura selecionada (French & Blanz-Jr. 1974). Estes compostos foram divididos em cinco séries, a saber: séries A, B, C, D, e E contendo, respectivamente, 40, 39, 30, 23 e 22 compostos, na tentativa de tornar estas séries estruturalmente mais homogêneas. Para cada série, foram criados três grupos de treinamento e os respectivos grupos de teste (I, II e III), visando-se avaliar o poder de predição dos modelos gerados através das análises de QSAR/QSAR-3D. Para as análises de QSAR clássico, foram utilizados como variáveis independentes, os descritores mais relevantes gerados através do programa DRAGON e, pré-selecionados por PLS. Considerando-se a ausência de informações sobre a estrutura cristalográfica da enzima RNR do M. tuberculosis, os estudos de QSAR-3D foram iniciados empregando-se metodologias propostas em CoMFA e, em CoMSIA, implementadas no programa SYBYL. Além destas, foi realizada a modelagem por homologia da RNR do M. tuberculosis, utilizando-se o programa WHATIF. Para as abordagens CoMFA e CoMSIA as geometrias otimizadas através do método semi-empírico AM1 foram alinhadas átomo-a-átomo e, através da similaridade dos respectivos campos estéricos e eletrostáticos, utilizando-se o programa SEAL. Nos dois procedimentos a geometria do composto não substituído, um dos mais ativos na série, foi utilizada como molde considerando-se a ausência de informações sobre a conformação bioativa. A modelagem da RNR por homologia foi realizada utilizando-se como molde as estruturas cristalográficas, respectivamente, do C. ammoniagenes (código PDB 1KGN) e da S. typhimurium (código PDB 1R2F), sambas apresentando valores de identidade superior a 65%. Mais recentemente foram publicados os dados cristalográficos para a cadeia beta (subunidade menor) da RNR do M. tuberculosis (código PDB 1UZR). Os modelos CoMFA e CoMSIA gerados apresentaram valores aceitáveis para os coeficientes de correlação de predição, com altos valores para os coeficientes de correlação ajustados e baixos valores para os erros padrões. Os melhores modelos CoMFA e CoMSIA foram obtidos considerando o grupo com substituintes apenas na posição 5 do anel piridínico. Razoáveis coeficientes de correlação de predição para os modelos CoMSIA com altos coeficientes de correlação de ajuste e baixos valores para os erros padrões forma obtidos. Os mapas de contorno gerados em CoMFA e CoMSIA sugerem que grupos aceptores de ligações de hidrogênio próximos ao nitrogênio do anel piridínico deverá aumentar o valor da atividade inibitória. Esta observação está em boa concordância com os dados da literatura, na qual a formação de um complexo entre a TSC e o íon Ferro foi sugerido para a inibição da RNR. Estes estudos deverão permitir um melhor entendimento sobre as características estruturais desta classe de TSC inibidoras da RNR, como agentes antitumorais, em termos dos campos estéricos, eletrostáticos, hidrofóbico, doador e aceptor de ligações de hidrogênio, bem como a contribuição para o desenvolvimento racional de novos inibidores para esta importante enzima. Adicionalmente, dois compostos preparados em nosso laboratório, demonstraram atividade frente o M. tuberculosis, em testes realizados in vivo. / Tuberculosis is an illness caused by Mycobacterium tuberculosis. Data from World Health Organization (WHO) estimates, that about 2-3 millions of human population died by Mycobacterium tuberculosis infection and that during the next 15 years about 1 billion will be infected and 35 million will certainly die. Although, in the clinic it was found several antiTBdrugs, these numbers will increase due several reasons including M. tuberculosis resistant strains. It has been stressed the importance of novel medicines and/or alternative biological targets research projects. It is known that Ribonucleotide reductase (RNR), is an enzyme that catalyses the rate limiting step in the de novo synthesis of dNTPs, reducing the ribonucleoside 5\'-diphosphates to the corresponding deoxyribonuc1eoside 5\' -diphosphates. RNR has a critical role in the DNA synthesis and, hence, cell division. This key enzyme, that shows 16% homology when compared with mammals RNR, is a potential target for drug design of cell growth inhibitors, with potential application in cancer therapy, antimalaria and trypanosome chemotherapy. It is known that different types of compounds or species by means of different mechanism pathways can show RNR inhibition, including &#945;-(N)-heterocyclic carboxaldehydes thiosemicarbazones that are one of the most potent classes of RNR inhibitors. More than that, some of them, that shows activity against M. tuberculosis seems to follow the same mechanism pathways proposed to the thiosemicarbazones tumor cells activity that means, that they probably are RNR inhibitors. In this study, a series of 40 &#945;-(N)-2-formyl-pyridine thiosemicarbazone derivatives tested against RNR of H.ep.-2-cells (human epidermoid carcinoma), taken from selected literature (French & Blanz-Jr. 1974), has quantitatively analyzed by means of several QSAR/3D-QSAR approaches. These compounds were divided into 5 individual subsets, namely A, B, C, D, and E, having 40, 39, 30, 23 e 22 compounds, respectively. This procedure has been done in order to achieve more structurally homogeneous subsets. For each set, three individual training and test sets (I,II and III) have been created in order to evaluate the predictivity power of the generated QSAR/3D-QSAR models. QSAR analysis have been done using descriptors generated by DRAGON program that have been further pre-selected by PLS procedures. Considering that crystallographic data of RNR M. tuberculosis are not available in the literature, 3D-QSAR studies have been done these applying, initially, CoMFA and CoMSIA approaches, implemented in SYBYL. Homology model studies have been performed with WHATIF program CoMFA e CoMSIA approaches used optimized geometry obtained by semi-empirical AM1 methods that have been aligned by two different methods. Rigid alignment, in which the compounds were fitted atom-by-atom onto a template, based on the root mean square fit. The N(l) and C(2) atoms of the pyridine moiety and the heavy atoms of thiosemicarbazone backbone of TSC were used as template structure. (2) Field based, in which the steric and electrostatic fields, generated by the SEAL program were considered in the alignment. In both procedures the unsubstituted 2-formylpyridine thiosemicarbazone in its syn conformation, has been taken as template. Homology RNR models were done using as template crystallographic data of ammoniagenes (1KGN) and S. typhimurium (1R2F) as template, respectively, with identity larger than 65%. More recent1y new crystallographic data have been published for the beta chain (smaller subunity) of RNR do M. tuberculosis (1UZR). CoMFA and CoMSIA generated models showed acceptable predictive correlation coefficients with high fitted correlation coefficients and low standard errors. Betler CoMFA and CoMSIA models have been derived considering a homogeneous subset of TSC substituted only at 5-position in pyridine ring. Reasonable predictive correlation coefficients for CoMSIA models with high fitted correlation coefficients and very low standard errors were obtained. The derived CoMFA and CoMSIA countour maps suggested that a hydrogen bond acceptor near the nitrogen pyridine ring could enhance inhibitory activity value. This observation is in good agreement with literature, in which a complex formation between TSC and iron ion has been suggested, to RNR inhibition. These studies are expected to enhance the understanding of the structural features of this class of TSC-RNR inhibitors as antitumor agents in terms of steric, electrostatic, hydrophobic and hydrogen donor and acceptor fields as well as to contribute to rational design of inhibitors of this key enzyme. Additionally, two compounds that have been prepared by us showed activity against M. tuberculosis using in vivo test system.