Made available in DSpace on 2016-04-28T20:15:25Z (GMT). No. of bitstreams: 1 2007-Fabiola do Nascimento Correa.pdf: 1170487 bytes, checksum: 12941bc2fa78c4d327c2059d03bf1c5c (MD5) Previous issue date: 2007-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Rearing buffaloes in Brazil has been increasing notoriously and leaving of being a simple activity to justify the use of poor fertility lands. Right now the rearing buffalo is considered a promissory lucrative activity, but the information about sanitary aspects to improve buffaloes health are not consistent. Borreliosis is a systemic infectious disease caused by many Borrelia species, which have a cosmopolitan distribution, and affects various species of domestic and wild animals, including human beings. Borrelia theileri is the specie more commonly reported in ruminants. However, these animals can also be infected by B. burgdorferi sensu lato and B. coriaceae, which cause Lyme borreliosis and abortion epizootic bovine, respectively. Meanwhile, there are no seroepidemiologic studies of borreliosis in buffaloes. The aim of this study was to know the frequency of anti- B. burgdorferi homologous antibodies in buffaloes serum samples proceeding from Castanhal, Santa Isabel, Nova Timboteua and Santar?m Novo, in the continental part of Par? state and Cachoeira do Arari in Maraj? Island were collected. Serums of 491 buffaloes were analyzed by indirect ELISA test. The serologic analysis of the samples showed that 412 serums (83.91%) were positive, without different statistical significance between positive animals from Maraj? Island 81.469% (2362/284) and from the continental part 86.96% (180/207). Was observed that buffaloes of these five municipalities studied have high antibodies frequency against spirochetes B. burgdorferi. The Correspondence Analysis test showed the formation of three different municipalities groups of according with seropositives animals number. The first group was formed by Cachoeira do Arari and Castanhal, the second by Nova Timboteua and Santar?m Novo. The third group was constituted by Santa Isabel only, which presented statisticment loss antibodies frequency than others municipalities. The high frequency of positives animals found can be explained by presence of tick Boophilus microplus and by the existence of report on Borrelia sp. infecting buffalo in the studied region. These facts suggest cross-reactivity between strain g39/40 of B. burgdorferi used as antigenic substratum and Borrelia species that infect buffaloes in Par? state. Despite of low specificity of indirect ELISA test used in this study, it is a good method to select e screen infected animals in studies about Borrelia sp. in buffaloes. / A cria??o de b?falos tem expandido no Brasil deixando de ser um simples elemento de ocupa??o de terras pouco f?rteis para tornar-se sin?nimo de produ??o pecu?ria rent?vel. No entanto, os pecuaristas e profissionais de sa?de carecem de informa??es consistentes sobre sanidade desses animais. A borreliose ? uma enfermidade sist?mica, infecciosa e cosmopolita, causada por microrganismos do g?nero Borrelia, que acometem diversas esp?cies de animais dom?sticos e silvestres, al?m do homem. A esp?cie de Borrelia mais comumente reportada em ruminantes ? B. theileri. Estes animais tamb?m podem ser infectados por B. burgdorferi sensu lato, agente da borreliose de Lyme e B. coriaceae que causa o aborto epizo?tico bovino. No entanto, n?o h? estudos soroepidemiol?gicos sobre borreliose em bubalinos. Com o objetivo de conhecer a freq??ncia de anticorpos hom?logos anti-B. burgdorferi de b?falos provenientes dos munic?pios Castanhal, Santa Isabel, Nova Timboteua e Santar?m Novo, na parte continental do estado do Par? e Cachoeira do Arari na Ilha de Maraj? foram coletados soros de 491 b?falos, os quais foram analisados por meio do teste ELISA indireto. A an?lise sorol?gica das amostras revelou que 412 soros (83,91%) foram positivos, n?o ocorrendo diferen?a estat?stica significativa entre os amimais positivos provenientes da Ilha de Maraj? 81,69% (232/284) e da por??o continental do estado 86,96% (180/207). Quanto ? freq??ncia de soropositivos por munic?pio estudado, a an?lise de correspond?ncia, indicou a forma??o de tr?s grupos distintos, o primeiro formado pelos munic?pios de Cachoeira do Arari e Castanhal, o segundo formado pelos munic?pios de Nova Timboteua e Santar?m Novo e o terceiro formado apenas pelo munic?pio de Santa Isabel. Este ?ltimo apresentou estatisticamente menor freq??ncia de anticorpos em rela??o aos outros quatro munic?pios. No entanto, foi observado que b?falos dos cinco munic?pios estudados possuem alta freq??ncia de anticorpos hom?logos contra espiroquetas B. burgdorferi. A alta freq??ncia de animais soropositivos encontrada pode ser explicada pela presen?a do carrapato Boophilus microplus e pela exist?ncia de relato sobre Borrelia sp. infectando b?falo na regi?o estudada. Estes fatos sugerem rea??o cruzada entre a cepa americana g39/40 de B. burgdorferi utilizada como substrato antig?nico e a esp?cie de Borrelia que infecta os b?falos no estado do Par?. Apesar da baixa especificidade do ELISA indireto usado neste estudo, este teste constitui-se em um bom exame para triagem e monitoramento de indiv?duos infectados por microrganismos do g?nero Borrelia.

Lyme disease is one of the most common vector-borne diseases around the world, and the numbers of reported cases are quickly rising. Ixodes ticks are the principal vectors, while Borrelia burgdorferi sensu lato genospecies are the etiological agents of the disease. Climate change, namely global warming, and habitat modification, namely forest fragmentation, are hypothesized to play an active role in this rise in reported cases. An analysis of the primary literature, specifically of studies focused on North America and Europe, was conducted in order to investigate these hypotheses. These studies show that global warming has precipitated a growth in tick populations as well as a northward tick migration, thereby increasing the risk of Lyme disease in emergent and endemic areas alike, for Borrelia spirochetes quickly infect naïve tick populations. Furthermore, published studies support the idea that forest fragmentation near human population centers has also increased the risk of Lyme disease in North America, for edge habitats provide suitable conditions for ticks and provide edible vegetation for the animals on which ticks feed, animals which also serve as hosts for B. burgdorferi sensu lato. In contrast, a decrease in fragmentation was found to facilitate tick invasion and establishment in Europe. These studies demonstrate that anthropogenic habitat modifications of varying types can affect ticks and their host populations and increase the risk of Lyme disease near human population centers. However, more research needs to be done to truly understand the different factors that are precipitating the rising number of cases of Lyme disease since there are significant interactions between climate change, habitat modification, and other drivers not examined here. Furthermore, understanding how these drivers function in specific geographic locations can help scientists and public officials tailor local public health measures appropriately. Finally, researchers and pharmaceutical companies must develop a safe, long-lasting, and effective vaccine against the Lyme disease spirochete, for there is not one currently available. Although easily treatable if diagnosed early, Lyme disease can progress to debilitating disease. Unfortunately, the risk of contracting this illness is currently rising and will continue to rise unless effective preventative measures are employed.

Lyme disease is the most commonly reported vector-borne disease in the United States. The causative agent of Lyme disease, can alter gene expression to enable survival in a diverse set of conditions, including the tick midgut and the mammalian host. External environmental changes can trigger gene expression in B. burgdorferi, and the data demonstrate that B. burgdorferi can similarly alter gene expression as a stress-response when it is treated with the antibiotic doxycycine. After treatment with the minimum bactericidal concentration (MBC) of doxycycline, a subpopulation can alter its phenotype to survive antibiotic treatment, and to host adapt and successfully infect a mammalian host. Furthermore, our data demonstrate that if a population is treated with the MBC of doxycycline, a subpopulation may alter its phenotype to adopt a state of dormancy until the removal of the antibiotic, whereupon the subpopulation can regrow. We demonstrate that the chance of regrowth occurring increases as a population reaches stationary phase, and present a mathematical model for predicting the probability of a persister subpopulation within a larger population, and ascertain the quantity of a persister subpopulation. To determine which genes are expressed as stress-response genes, RNA Sequencing analysis, or RNASeq, was performed on treated, untreated, and treated and regrown B. burgdorferi samples. The results suggest several genes were significantly different in the treated group, compared to the untreated group, and in the untreated and regrown group compared to the untreated group, including a 50S ribosomal stress-response protein, coded from BB_0786. The appendices discuss the theory and methods that were used in RNA Sequencing (RNASeq) analysis, and provide an overview of the database that was created for the B. burgdorferi transcriptome. Additional studies may demonstrate further how persister subpopulations form, and which genes can trigger a persister state in B. burgdorferi. / acase@tulane.edu

<p>Ticks are vectors for a larger number of viruses and bacteria than all other arthropod taxa, including mosquitoes. In Europe is it foremost Borrelia spirochetes and the Flavivirus Tick-borne Encephalitis virus that cause disease in humans. In this study, the tick species <em>Ixodes uriae</em> has been studied. <em>I.</em><em> uriae</em> have a circumpolar distribution in both hemisphere and can be found both in Arctic and Antarctica. I collected ticks from Magellanic penguins in south Chile and analyzed them to see if they carry <em>Borrelia</em> spirochetes or Flavivirus. Totally were 218 ticks collected from 165 controlled penguins. All ticks were collected from adult penguins and the parasitizing ticks were all found in the auditory meatus which is a new phenomena compared to earlier studies. Both <em>Borrelia</em> spirochetes and Flavivirus were found in the collected ticks using PCR techniques. This is an interesting result since not much research has been performed in this geographical area before. Until date there is only one species of <em>Borrelia</em> (<em>Borrelia</em> <em>garinii</em>) found in <em>I.</em><em> uriae</em> on the southern hemisphere and new Flavivirus is regularly found around the world.  Unfortunately we have not been able to determine species of the <em>Borrelia </em>spirochetes or Flavivirus so far but this work will be continued. <strong></strong></p>

Ticks are vectors for a larger number of viruses and bacteria than all other arthropod taxa, including mosquitoes. In Europe is it foremost Borrelia spirochetes and the Flavivirus Tick-borne Encephalitis virus that cause disease in humans. In this study, the tick species Ixodes uriae has been studied. I. uriae have a circumpolar distribution in both hemisphere and can be found both in Arctic and Antarctica. I collected ticks from Magellanic penguins in south Chile and analyzed them to see if they carry Borrelia spirochetes or Flavivirus. Totally were 218 ticks collected from 165 controlled penguins. All ticks were collected from adult penguins and the parasitizing ticks were all found in the auditory meatus which is a new phenomena compared to earlier studies. Both Borrelia spirochetes and Flavivirus were found in the collected ticks using PCR techniques. This is an interesting result since not much research has been performed in this geographical area before. Until date there is only one species of Borrelia (Borrelia garinii) found in I. uriae on the southern hemisphere and new Flavivirus is regularly found around the world.  Unfortunately we have not been able to determine species of the Borrelia spirochetes or Flavivirus so far but this work will be continued.

Thesis (M.S.)--Ball State University, 2009. / Title from PDF t.p. (viewed on Feb. 08, 2010). Includes bibliographical references (p. 81-94).

Borrelia burgdorferi is an elongated and helically shaped bacterium that is the causal agent of the tick-borne illness Lyme disease. The disease manifests with initial flu-like symptoms and, in many cases, the appearance of a skin rash called erythema migrans at the site of the tick bite. If left untreated the disease might cause impairment of various organs such as the skin, heart, joints and the nervous system. The bacteria have a parasitic lifestyle and are always present within a host. Hosts are usually ticks or different animals and birds that serve as reservoirs for infection. B. burgdorferi are unable to synthesize building blocks for many vital cellular processes and are therefore highly dependent on their surroundings to obtain nutrients. Because of this, porins situated in the outer membrane, involved in nutrient uptake, are believed to be very important for B. burgdorferi. Except for a role in nutrient acquisition, porins can also have a function in binding extracellular matrix proteins, such as integrins, and have also been implicated in bacterial adaptation to new environments with variations in osmotic pressure. P13 and P66 are two integral outer membrane proteins in B. burgdorferi previously shown to have porin activities. In addition to its porin function, P66 also has integrin binding activity. In this thesis, oligomeric structures formed by the P13 and P66 protein complexes were studied using the Black lipid bilayer technique in combination with nonelectrolytes. Initial attempts were also made to study the structure of P13 in Nanodiscs, whereby membrane proteins can insert into artificial lipid bilayers in their native state and the structure can be analyzed by electron microscopy. In addition, the role of P13 and P66 in B. burgdorferi osmotic stress adaptation was examined and also the importance and role of the integrin-binding activity of P66 in B. burgdorferi infections in mice. Using Black lipid bilayer studies, the pore forming activity of P13 was shown to be much smaller than previously thought, exhibiting activity at 0.6 nS. The complex formed by P13 was approximately 300 kDa and solely composed of P13 monomers. The channel size was calculated to be roughly 1.4 nm. Initial Nanodisc experiments showed a pore size of 1.3 nm, confirming the pore size determined by Black lipid bilayer experiments. P66 form pores with a single channel conductance of 11 nS and a channel size of 1.9 nm. The porin assembles in the outer membrane into a large protein complex of 420 kDa, containing exclusively P66 monomers. The integrin-binding function of P66 was found to be important for efficient bacterial dissemination in the murine host but was not essential for B. burgdorferi infectivity. Neither P13 nor P66 had an active role in osmotic stress adaptation. Instead, two p13 paralogs were up-regulated at the transcript level in B. burgdorferi cultured under glycerol-induced osmotic stress. / Borrelia burgdorferi är en bakterie med många unika egenskaper som orsakar sjukdomen Lyme borrelios. Borrelia kan idag lätt behandlas med antibiotika om sjukdomen upptäcks i ett tidigt stadium. Det är först om sjukdomen tillåts fortgå som symptom som nervsmärta och ansiktsförlamning kan uppstå och dessutom vara svåra att koppla till en Borrelia-infektion. Multiresistenta bakterier har blivit en stor del av vår vardag och även om Borrelia-bakterierna idag inte är resistenta mot flertalet antibiotika är det kanske speciellt viktigt, innan det är för sent, med forskning som kan leda till upptäckter av unika angreppsställen för nya läkemedel. Målet med denna avhandling var att studera hur två Borrelia proteiner, P13 och P66, ser ut, är uppbyggda och även vilken funktion de har. Dessa proteiner är tänkbara vaccinkandidater eftersom de sitter i yttre membranet hos bakterierna och sticker ut på ytan mot våra värdceller, vilket gör att vi reagerar mot dem vid en infektion. P13 och P66 är också viktiga kanaler för bakterierna vid upptag av näringsämnen och byggstenar från omgivningen. Ämnen som bakterierna inte kan producera själva. Pga. denna funktion är P13 och P66 tänkbara proteiner för blockering med ett läkemedel som skulle förhindra bakterien från att föröka sig i och med att de förlorar möjligheten att tillgodogöra sig näring. Detta i sin tur skulle leda till att vårt eget immunförsvar hinner rensa undan bakterierna innan infektionen blivit för stor och vi blivit sjuka. P66 har förutom porin funktionen även en adhesions funktion när proteinet kan binda integriner som sitter på olika typer av celler i vår kropp, bl. a. immunceller och epitelceller i våra blodkärl och vävnader. Den integrin bindande funktionen är viktig för bakterierna vid en infektion eftersom det gör det möjligt för bakterierna att binda till våra celler. Ett steg som är viktigt för att de senare ska kunna ta sig ut från blodkärlen till våra vävnader. P13 och P66 visade sig kunna bilda stora proteinkomplex i ytter membranet hos bakterierna med en storlek på 300 kDa respektive 420 kDa. De är inga specifika poriner som bara kan transportera en viss typ av molekyl med t.ex. en viss laddning, utan kan ombesörja upptaget av många olika typer av ämnen. Eliminering av p66 orsakade att ett annat adhesionsprotein, uppreglerades. En omplacering av ett normalt cytoplasmatiskt lokaliserat chaperon-protein till ytter-membranet hos bakterierna kunde också ses i frånvaro av P66. Chaperonet GroEL har i andra bakterier, bl. a. Helicobacter pylori, bakterien som orsakar magsår, beskrivits som ett protein som kan förflytta sig till ytan av bakterierna och där ha en liknande funktion som P66, dvs. att binda extracellulära matrisprotein. Förändringen i uttryck av adhesionsproteinet och förflyttningen av chaperonet till membranet var en följd av p66-eliminering och mest troligt ett sätt för bakterierna att komplettera den förlorade integrinbindande funktionen av P66. Det har tidigare visats att poriner är involverade i skyddet mot osmotisk stress i andra bakterier. Denna funktion hos P13 och P66 i Borrelia kunde inte ses när bakterier utsattes för osmotisk stress med glycerol, som orsakar en form av membranstress. Däremot kunde vi med hjälp av transkriptomanalys se att Borrelia-bakterier uppreglerade transkriptionen av två paraloger till P13 vid hyper-osmotisk stress. Borrelia bakteriens användning av dessa paraloga proteiner har tidigare trotts ske enbart i frånvaro av ett funktionellt P13 protein. Nu visade det sig att P13-paraloger har en egen funktion även i närvaro av P13, nämligen att vara involverade i regleringen av hyperosmotisk stress och därmed skydda bakterierna i denna stressituation. Andra gener som påverkades av osmotisk stress med glycerol var gener för stressfaktorer och pumpar i inre membranet hos bakterien.

The two main causes of child mortality and morbidity in Africa are malaria and invasive bacterial diseases. In addition, co-infections in sub-Saharan Africa are the rule rather than the exception. However, not much is known about the host-pathogen interaction during a concomitant infection or how it affects the outcome of disease. In order to study the immunological responses during malaria and bacterial co-infections, we established a co-infection mouse model. In these studies we used two pathogenic bacteria found in malaria co-infected patients: Streptococcus pneumoniae and Relapsing fever Borrelia duttonii. Hosts co-infected with malaria and Borrelia showed greatly increased spirochetal growth but low parasite densities. In addition, the co-infected hosts presented symptoms of experimental-cerebral malaria, in an otherwise unsusceptible mouse model. This was found to be a consequence of a dysregulated immune response due to loss of timing and control over regulatory mechanisms in antigen presenting cells thus locking the host in an inflammatory response. This results in inflammation, severe anemia, internal organ damage and pathology of experimental cerebral malaria. On the other hand, in the malaria - S. pneumoniae co-infection model we found that co-infected hosts cleared the bacterium much more efficiently than the single infected counterpart. This efficiency of clearance showed to be neutrophil dependent. Furthermore, in vitro studies revealed that neutrophils isolated from malaria-infected hosts present an altered migratory effect together with a significantly increased capacity to kill S. pneumoniae. This suggests that a malaria infection primes neutrophils to kill S. pneumoniae more efficiently. Furthermore, a study was carried out on plasma samples from Rwandan children under the age of five, on which a full metabolomics profile was performed. We showed that these children could be divided in different disease categories based on their metabolomics profile and independent of clinical information. Additionally, the mild malaria group could further be divided in two sub-groups, in which one had a metabolomic profile resembling that of severe malaria infected patients. Based on this, metabolite profiling could be used as a diagnostic tool to determine the distinct phase, or severity of a malaria infection, identify risk patients and provide helpful and correct therapy.

The spirochete Borrelia hermsii avoids the immune response of its mammalian host through multiphasic antigenic variation. Serotype specificity is determined by Variable major proteins (Vmp), in the outer membrane. Through a non reciprocal recombination between linear plasmids, a formerly silent vmp gene replaces another vmp gene at a telomeric expression locus downstream from a common expression site. B. hermsii before and after the switch from serotype 7 to serotype 21, was examined in detail. The nucleotide sequence of the vmp7 and vmp21 genes and flanking regions was determined. The vmp7 and vmp21 are 77% identical in their coding sequence, and the deduced translation products are 63% identical. No antigenic cross reactivity is observed between Vmp7 and Vmp21. This suggests a folding of the proteins in which the similar regions are buried, and not exposed when it is presented at the bacterial surface. Vmp7 and Vmp21 have consensus sequences of prokaryotic lipoproteins and are processed as such when expressed in E. coli. The 5' regions of silent and expressed vmp7 and vmp21 were compared. Silent and active vmp7 and vmp21 genes shared a block of homologous sequence at their 5' ends. Sequences upstream of silent vmp7 and vmp21 genes lacked a promoter and differed substantially from each other. In this antigenic switch a vmp gene was activated by a recombination event which placed it downstream of a promoter. The vmp gene promoter is preceded by a poly(dT dA) ran and three imperfectlyrepeated elements of 2 kb. Each of the 2 kb repeats contains inverted repeats of approximately 0.2 kb at their termini. There is no evidence of the presence of similar elements elsewhere in the genome of B. hermsii. One or more of these elements may stimulate vmp gene switch or expression. The African relapsing fever species Borrelia crocidurae and the American species B. hermsii display many similarities. In both species the vmp genes are localised to linear plasmids, and the vmp genes are activated on the transcriptional level. The nucleotide sequence of their expression sites, however, are not related. Still, the possibility that the switch is mechanistically similar in B. crocidurae and B. hermsii, cannot be ruled out. The binding of B. crocidurae causes aggregation of erythrocytes around the spirochete. The aggregation is reminiscent of the erythrocyte rosetting seen in malarial infections. The erythrocytes at the B. crocidurae surface may protect them from clearance by the host. Thus, the rosetting may constitute an additional mechanism in B. crocidurae for the evasion of the immune reaction. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1994, härtill 5 uppsatser.</p> / digitalisering@umu

Uninfected larvae of Ixodes dammini, lxodes cookei, and Amblyomma americanum were fed on hamsters that had been injected intraperitoneally with a 0.5.ml sample of Borrelia burgdorferl (2.5 X 107 spirochetes per ml) 21 days earlier. A total of 108 of these larvae comprised of 36 1. dammini, 36 i. cookei, and 36 A. americanum were aseptically dissected and examined by darkfield and immunofluorescent microscopy for the presence of B. burgdorferl within 48 hours of feeding on the B. burgdorferi infected hamsters. The removal and examination of the midgut diverticula revealed that 32/36 (88.9%) of the l. dammini larvae contained B. burgdorferl. Only 5/36 (13.9%) of the l. cookei larvae and 7/36 of the A. americanum larvae harbored spirochetes in their midgut diverticula.A portion of the nymphs that molted from the above larvae were also dissected and examined by darkfleld and indirect immunofluorescent techniques. Borrelia burgdorferi were observed in the midgut diverticula of 94/107 (87.8%) of the l. dammini nymphs. None of the 30 (0%) l. cookei nymphs examined were found positive for spirochetes and only 1/60 (1.7%) of the A. americanum nymphs was found positive for B. burgdorrerl.A total of 83 lL dammini, 53 A. americanum, and 161. cookei nymphs reared from larvae that fed to repletion on hamsters infected with B. burgdorrerl were allowed to feed on uninfected hamsters to assess transmission of B. burgdorrerl. Transmission was demonstrated only by the l. dammlnl nymphs. The findings of this study suggest that it is extremely unlikely that l. cookei can serve as a vector for B. burgdorrerl, but do not rule out completely the possibility that A. americanum may be able to maintain B, burgdorrerl infections transstadially and, under certain conditions, transmit the organisms to vertebrate hosts. / Department of Physiology and Health Science