Browsing by Subject "virulenssi"
Now showing items 1-5 of 5
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(University of HelsinkiHelsingin yliopistoHelsingfors universitet, 2007)Koagulaasi-negatiiviset stafylokokit (KNS) ovat gram-positiivisia bakteereita. Lehmän utaretulehduksista eritettyjä KNS-lajeja on noin kaksikymmentä. Yleisimpiä lajeja mastiitin aiheuttajina ovat S. simulans, S. chromogenes, S. hyicus ja S. xylosus. Koagulaasinegatiivisia stafylokokkeja on pidetty vähämerkityksellisinä utaretulehduksen aiheuttajina. Useat tutkimukset osoittavat kuitenkin, että KNS:t ovat monessa maassa yleisin utaretulehduksen aiheuttaja. KNS:t eivät ole homogeeninen ryhmä, vaan koostuvat useista eri lajeista ja kannoista. Niiden aiheuttamat kliiniset oireet vaihtelevat. KNS- mastiitin patogeneesistä ja siihen vaikuttavista tekijöistä tiedetään hyvin vähän. Tämän työn tarkoituksena oli kirjallisuuskatsauksen muodossa kartoittaa mitä KNS:ien virulenssitekijöitä on tutkittu ja mitä menetelmiä käytetty. Jatkossa on suunnitteilla tutkimus, jossa selvitetään mitä virulenssitekijöitä yleisemmin utaretulehduksissa esiintyvät KNS:t kantavat genomissaan ja onko eri lajien välillä eroja. Tutkittavat geenit voidaan jakaa kahteen ryhmään: bakteerin tarttumiseen eli adheesioon ja kolonisaatioon liittyviä proteiineja koodaavat geenit (esim. biofilmin muodostus (ica ja bap), fibrinogeenia, kollageenia ja fibronektiinia sitovat proteiinit), ns. klassiset virulenssitekijät, kuten hemolysiinit, leukosidiinit, enterotoksiinit A-J, TSST-1, sekä mikrobilääkeresistenssiä koodaavat geenit mecA ja blaZ. Toistaiseksi ei löytynyt näyttöä siitä, että joku KNS -laji olisi virulentimpi kuin muut. Yksittäisissä tutkimuksissa on epäilty jonkun tutkimuksessa mukana olleen KNS:n olevan virulentimpi kuin muut yhden virulenssitekijän löytymisen perusteella. Useimmissa tutkimuksissa oli tutkittu yhtä tai kahta virulenssitekijää ja tutkimuksissa on ollut mukana vain muutama KNS -laji ja -kanta. Kirjallisuuskatsauksen yhteenvetona todettakoon, että KNS:ien virulenssitekijöitä ei ole tutkittu kovinkaan paljoa maailmalla, ja tehdyt tutkimukset ovat olleet suppeita. Stafylokokeista S. aureuksen virulenssitekijöitä sen sijaan on tutkittu huomattavasti enemmän. Virulenssitekijöiden tutkimiseen on enimmäkseen käytetty molekyylibiologisia menetelmiä.
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(2008)Pectobacterium atrosepticum is a bacterium that is a causative agent of blackleg and soft rot of potatoes at temperate climate zone. The genome of P. atrosepticum was sequenced at 2004. Sequence revealed six vgrG genes, and five of them are predicted to encode for proteins having uncharacterized function. VgrG proteins are found to be secreted by a novel secretion system called type VI secretion. They are also suggested to be a part of the type VI secretion translocon. VgrG proteins are presumed to be cytotoxic to the hosts of pathogenic bacteria. A mutation in a vgrG gene attenuates virulence of a few human and animal pathogenic bacteria. The aim of this study was to find out the affect of vgrG genes to the virulence of P. atrosepticum. In this study, transcription of vgrG genes of P. atrosepticum was measured by a real-time reverse transcriptase PCR. P. atrosepticum was grown in a virulence gene inducing conditions. Most active vgrG genes were deleted by Red recombinase using method. Each mutant strain carried deletion in one of the active vgrG genes. Virulence of the mutant strains was assessed on potato tubers, according to their ability to rot tissue. Three vgrG genes were induced in bacteria when they grew in the host tissue. vgrG genes were most active in samples, which were taken from stems 96 hours after inoculation. One mutant strain was slightly more virulent (P = 0,0077). Two strains were slightly less virulent, but results were not statistically significant (P > 0,05). According to the profile of transcription, vgrG genes might be novel virulence genes. However, virulence essays did not support that possibility. In fact, one of the mutations increased the virulence of P. atrosepticum. It is possible that other vgrG genes cover the function of the deleted gene. There is no clear evidence that vgrG genes or the type VI secretion are novel virulence determinants of P. atrosepticum. Perhaps they have some other function in the active stage of the infection. Example, they might protect P. atrosepticum from competitive microbes in the rotten potato tissue.
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(2020)Constantly increasing level of bacteria becoming resistant to clinically relevant antibiotics challenges the modern medical achievements made over the past century. In global scale, one of the most significant information gaps concerning the occurrence of resistant bacteria is located in West African countries. Klebsiella pneumoniae and Escherichia coli strains resistant to 3rd generation cephalosporins and carbapenems are a major risk to public health through infections with limited or no available treatment options. The resistance to these antibiotics among Enterobacteriaceae is mainly mediated by hydrolyzing enzymes such as extended-spectrum beta-lactamases (ESBL). The focus of this thesis is to study the genes encoding these enzymes and other resistance factors found in K. pneumoniae and E. coli isolated from human stool and waste water samples in Burkina Faso and Mali. Tree Enterobacteriaceae isolates were selected for whole genome sequence (WGS) analysis based on their phenotypic resistance profiles defined by disk diffusion method. Reads were assembled to draft genomes and the genomes were studied for their antibiotic resistance genes, virulence genes and their associations to mobile genetic elements found in these isolates’ genomes. Additionally a pan-genome was created to investigate species specific features of K. pneumoniae and their role in heavy load of antibiotic resistance genes among these isolates. Pan-genome consisted of two genomes sequenced in this study and 12 genomes from the publically available database. 16-month old Burkinabe child was a carrier of one ESBL-producing K. pneumoniae (isolate Burkina_1) and one ESBL-positive E. coli along with the resistance to multiple other antibiotics. With genome wide analysis the K. pneumoniae strain could be described as sequence type (ST) 45 representing, multidrug resistant and ESBL-gene CTX-M-15 carrying strain with highly similar virulence gene profile to strains previously described as pathogenic K. pneumoniae causing neonatal sepsis. K. pneumoniae isolated from the stool sample of an adult living in Burkina Faso was found to be multidrug resistant, though non-ESBL-producer strain (isolate Burkina_2). The isolate showed no similarity to any previously described sequence type. CTX-M-15 encoding E. coli of ST38 (isolate Mali_1) carried by Malian child showed resistance to five different classes of antibiotics in addition to the 3rd generation cephalosporins. At the same time the isolate showed hybrid virulence gene profile with virulence genes associated to many different E. coli pathotypes including neonatal meningitis causing E. coli (NMEC). The exceptional plasticity of K. pneumoniae genome could be recognized as one of the putative explanations for the high number of resistance genes found among the isolates studied in this work. Antibiotic resistance genes were found to be associated to mobile genetic elements (MGE) and as the genetic plasticity is caused by the acquisition of external genetic material via MGEs such as plasmids, this can lead to indirect accumulation of resistance genes in these genomes. The results in this thesis work show alarming examples of pathogens that potentially cause severe infections, have extremely narrow or no treatment options and are carried by infants. These findings are in line with the few data about the level of faecal carriage of ESBL-producing strains by people in Burkina Faso and Mali reported previously.
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(2020)Constantly increasing level of bacteria becoming resistant to clinically relevant antibiotics challenges the modern medical achievements made over the past century. In global scale, one of the most significant information gaps concerning the occurrence of resistant bacteria is located in West African countries. Klebsiella pneumoniae and Escherichia coli strains resistant to 3rd generation cephalosporins and carbapenems are a major risk to public health through infections with limited or no available treatment options. The resistance to these antibiotics among Enterobacteriaceae is mainly mediated by hydrolyzing enzymes such as extended-spectrum beta-lactamases (ESBL). The focus of this thesis is to study the genes encoding these enzymes and other resistance factors found in K. pneumoniae and E. coli isolated from human stool and waste water samples in Burkina Faso and Mali. Tree Enterobacteriaceae isolates were selected for whole genome sequence (WGS) analysis based on their phenotypic resistance profiles defined by disk diffusion method. Reads were assembled to draft genomes and the genomes were studied for their antibiotic resistance genes, virulence genes and their associations to mobile genetic elements found in these isolates’ genomes. Additionally a pan-genome was created to investigate species specific features of K. pneumoniae and their role in heavy load of antibiotic resistance genes among these isolates. Pan-genome consisted of two genomes sequenced in this study and 12 genomes from the publically available database. 16-month old Burkinabe child was a carrier of one ESBL-producing K. pneumoniae (isolate Burkina_1) and one ESBL-positive E. coli along with the resistance to multiple other antibiotics. With genome wide analysis the K. pneumoniae strain could be described as sequence type (ST) 45 representing, multidrug resistant and ESBL-gene CTX-M-15 carrying strain with highly similar virulence gene profile to strains previously described as pathogenic K. pneumoniae causing neonatal sepsis. K. pneumoniae isolated from the stool sample of an adult living in Burkina Faso was found to be multidrug resistant, though non-ESBL-producer strain (isolate Burkina_2). The isolate showed no similarity to any previously described sequence type. CTX-M-15 encoding E. coli of ST38 (isolate Mali_1) carried by Malian child showed resistance to five different classes of antibiotics in addition to the 3rd generation cephalosporins. At the same time the isolate showed hybrid virulence gene profile with virulence genes associated to many different E. coli pathotypes including neonatal meningitis causing E. coli (NMEC). The exceptional plasticity of K. pneumoniae genome could be recognized as one of the putative explanations for the high number of resistance genes found among the isolates studied in this work. Antibiotic resistance genes were found to be associated to mobile genetic elements (MGE) and as the genetic plasticity is caused by the acquisition of external genetic material via MGEs such as plasmids, this can lead to indirect accumulation of resistance genes in these genomes. The results in this thesis work show alarming examples of pathogens that potentially cause severe infections, have extremely narrow or no treatment options and are carried by infants. These findings are in line with the few data about the level of faecal carriage of ESBL-producing strains by people in Burkina Faso and Mali reported previously.
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(University of HelsinkiHelsingin yliopistoHelsingfors universitet, 2012)Streptococcus uberis on gram-positiivinen kokki, jonka merkitys lehmien utaretulehdusten aiheuttajana on kasvanut viime aikoina. Se on opportunistinen bakteeri, jonka aiheuttama taudinkuva voi olla hyvinkin vakava. Toisaalta se aiheuttaa myös paljon subkliinisiä infektioita. S. uberis –bakteerin virulenssitekijöitä ei ole kovin paljoa tutkittu. Kirjallisuuden perusteella mahdollisia virulenssitekijöitä ovat hyaluronihappokapseli, neutrofiili-toksiini, M- ja R-proteiinien kaltaiset proteiinit, plasminogeenin aktivointi, laktoferriiniä sitovat proteiinit (SUAM), metal transporter uberis A (mtuA), hyaluronidaasi ja niin kutsuttu uberis-tekijä. Mikrobilääkeresistenssi on maailmanlaajuisesti kasvava ongelma. Suomessa tilanne on vielä melko hyvä, mutta meilläkin resistenssi on viime aikoina lisääntynyt. S. uberis –bakteerilla on Suomessa todettu resistenssiä esimerkiksi erytromysiiniä ja oksitetrasykliiniä vastaan, ulkomailla resistenssi on yleistä myös muita mikrobilääkkeitä vastaan. Tutkimuksessani määritin ja vertasin kapseligeenien hasA ja hasC sekä laktoferriiniä sitovaa proteiinia (SUAM) koodaavan sua-geenin esiintymistä subkliinistä ja kliinistä utaretulehdusta aiheuttaneiden S. uberis –kantojen välillä. SUAM:in esiintyvyydessä ei havaittu merkittävää eroa (p > 0,05). Sen sijaan kapselia koodaavia geenejä esiintyi useammin kliinistä kuin subkliinistä utaretulehdusta aiheuttaneilla kannoilla (p < 0,01), mikä viittaisi kapselin mahdollisesti olevan merkittävä tekijä taudinaiheutuksen kannalta. Lisäksi tutkin antibioottiresistenteiltä kannoilta erytromysiiniresistenssiä koodaavan mefA-geenin sekä tetrasykliiniresistenssiä koodaavien tetK-, tetL-, tetM-, tetO- ja tetS-geenien esiintymistä. Erytromysiiniresistenssin aiheuttaja jäi tuntemattomaksi, sillä mefA-geeniä ei löytynyt yhdeltäkään tutkituista kannoista. Tetrasykliiniresistenssigeeneistä 70,4:ltä %:lta tutkituista kannoista löytyi tetL-geeni, lisäksi 14,8:lta %:lta löytyi tetM-geeni. TetM esiintyi yhdessä tetL-geenin kanssa. 29,6:lla %:lla kannoista resistenssigeeni jäi tuntemattomaksi.
Now showing items 1-5 of 5