The tick Ixodes ricinus is one of the most important vectors of human pathogens like the spirochete Borrelia burgdorferi and TBE virus. The effect of repeated infestations with pathogen-free I. ricinus nymphs and the effect of vaccination with ticks saliva of the tick I. ricinus on the transmision Borrelia burgdorferi and on the production of the cytokines were studied. The draining lymph nodes of C3H/HeN mice were examined for IL-2, IFN-? and IL-10 production by antigen capture enzyme-linked immunosorbent assay. Spirochete load in skin, heart and bladder was examined by real-time polymerase chain reaction.

The thesis focuses on the investigation of the role of tick salivary components in the course of Lyme disease in mice. It includes studies on the saliva-facilitated transmission of Borrelia burgdorferi in vivo and the effect of tick cysteine protease inhibitors (cystatins) both on murine immune cells and the transmission of B. burgdorferi spirochetes in mice. The thesis also reveals practical applications of salivary cystatins for the development of anti-tick vaccine and the application of the pharmacological action of a tick salivary cystatin for the therapy of the disease symptoms in a mouse model of experimental asthma.

Die vorliegende epidemiologische Studie umfaßte 153 Hunde aus der Nähe von Athen, Griechenland. Um die Prävalenz arthropogener Infektionen abschätzen zu können, wurden Serumproben auf Antikörper gegen Leishmania infantum, Ehrlichia canis und Borrelia burgdorferi sowie auf Antigene von Dirofilaria immitis überprüft. Blutausstriche wurden auf das Vorkommen von Babesia canis und Hepatozoon canis untersucht. Außerdem wurden von den Hunden abgesammelte Zecken bestimmt. Bei 126 Hunden erfolgte eine klinische Allgemeinuntersuchung. Des weiteren wurden die serologischen Testverfahren ELISA und IFAT für den Nachweis von Antikörpern gegen Borrelia burgdorferi miteinander verglichen. Insgesamt waren 94 (61,4 %) der 153 untersuchten Hunde infiziert, 63 (41,2 %) Hunde wiesen Antikörper gegen Ehrlichia canis auf. Infektionen mit Borrelia burgdorferi wurden im ELISA insgesamt bei 43 (28,1 %), im IFAT bei 35 (22,9 %) Tieren nachgewiesen. Außerdem konnte bei 28 (18,3 %) Tieren eine Infektion mit Leishmania infantum und bei 20 (13,1 %) mit Dirofilaria immitis gefunden werden. Ein Nachweis von Babesia canis im Blutausstrich gelang bei vier (2,6 %) Hunden, von Hepatozoon canis nur bei einem (0,7 %) Hund. Für keinen der untersuchten Infektionserreger konnten signifikante Alters-, Geschlechts- oder Rasseprädispositionen festgestellt werden. Die Infektionsraten mit Leishmanien, Babesien, Dirofilarien und Borrelien waren bei den im Tierheim lebenden Hunden geringer als bei den Streunern. Vierundvierzig (28,8 %) der 153 getesteten Hunde waren gleichzeitig mit zwei, drei oder vier durch Arthropoden übertragene Erregerarten infiziert. Zweifachinfektionen kamen bei 29 (19,0 %), Dreifachinfektionen bei sieben (4,6 %) und Vierfachinfektionen bei acht (5,2 %) Tieren vor. In der Regel war das Risiko für Hunde, einen Erreger zu beherbergen, höher, wenn bereits eine Infektion mit einem anderen Erreger vorhanden war. Insgesamt waren 28 (18,3 %) Streuner und 15 (9,8 %) Tierheimhunde von Mehrfachinfektionen betroffen. Von den 94 mit mindestens einem der berücksichtigten Erreger infizierten Hunden wurden 75 (79,8 %) einer klinischen Untersuchung unterzogen. Dreiunddreißig (44,0 %) dieser Tiere zeigten Krankheitserscheinungen. Bei 21 (67,7 %) der 31 klinisch untersuchten und mit mehreren Erregern gleichzeitig infizierten Hunde konnten Krankheitssymptome diagnostiziert werden. Von 44 klinisch untersuchten Tieren, die nur mit einem Erreger infiziert waren, zeigten 12 (27,3 %) klinische Symptome. Die diagnostizierten Symptome waren bei allen untersuchten Erregern von einer starken Variabilität geprägt. Die Übereinstimmung der Testsysteme IFAT und ELISA für den Nachweis von Antikörpern gegen Borrelia burgdorferi lag bei 92,2 %. Die Testverfahren sind damit als nahezu gleichwertig anzusehen, wobei der ELISA etwas sensitiver zu sein scheint. Eine Infektionsgefahr besteht demnach in Griechenland vor allem für Ehrlichia canis, Borrelia burgdorferi und Leishmania infantum, aber auch für Infektionen mit Dirofilaria immitis besteht ein Risiko. Infektionen mit Babesia canis kommen in dieser Region seltener vor, es muss jedoch insbesondere bei geschwächten oder in Deutschland geborenen und somit hochempfänglichen Tieren mit Erkrankungen gerechnet werden. Hepatozoon canis ist nach den vorliegenden Ergebnissen in der Umgebung von Athen von geringer Verbreitung. Ein Rückschluß vom klinischen Bild auf das Vorliegen einer Infektion mit einem bestimmten Erreger kann aufgrund der Variabilität der Symptome in Verbindung mit häufig auftretenden Mehrfachinfektionen nicht gezogen werden. Die Möglichkeit von gleichzeitig vorliegenden Infektionen sollte in der Diagnostik und Therapie unbedingt berücksichtigt werden. Gute Haltungsbedingungen und eine tierärztliche Überwachung und Prophylaxe reduzieren offensichtlich die Inzidenz von Infektionen mit arthropodenübertragenen Erregern. Die überwiegende Anzahl (449 von 457 Exemplaren) der bestimmten Zecken gehörte der Art Rhipicephalus sanguineus an. Es wurden sechs Nymphen und 443 adulte Tiere gefunden. Die adulten Tiere verteilten sich auf 243 männliche und 200 weibliche Zecken. Bei acht Zecken handelte es sich um weibliche Ixodes ricinus. Bei aus Griechenland stammenden Hunden wie auch bei Tieren, die sich reisebegleitend dort aufgehalten haben, muss mit einer starken Infestation mit Rhipicephalus sanguineus gerechnet werden. Da Rhipicephalus sanguineus der Vektor für Ehrlichia canis, Babesia canis vogeli und Hepatozoon canis ist, sollten Prophylaxemaßnahmen eine geeignete Zeckenbekämpfung einbeziehen. / 153 dogs from the environs of Athens, Greece, were surveyed for tick infestation and arthropod borne infections. Serology was performed for Leishmania infantum, Dirofilaria immitis, Ehrlichia canis and Borrelia burgdorferi and bloodfilms were microscopically examined for Babesia canis and Hepatozoon canis. Ticks collected from the dogs were differentiated. 126 dogs underwent clinical examination. Suitability of an indirect immunofluorescent antibody assay (IFAT) and an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of antibodies to Borrelia burgdorferi was compared. Altogether 94 (61.4 %) dogs were infected with an arthropod borne pathogen, 63 (41.2 %) produced antibodies to Ehrlicha canis. ELISA detected Borrelia burgdorferi infection in 43 (28.1 %) dogs while IFAT was positive in 35 (22.9 %). 28 (18.3 %) dogs were infected with Leishmania infantum and 20 (13.1 %) with Dirofilaria immitis. Babesia canis was found in blood smears of four (2.6 %) dogs, Hepatozoon canis was detected only in one case (0,7 %). No association was found between the breed, age or sex of the dogs and any of the tested pathogens. The rate of infection with Leishmania, Babesia, Dirofilaria or Borrelia was lower in dogs living in the animal shelter than in those living as strays. 44 (28.8 %) of the 153 dogs examined were infected concurrently with two (n = 29, 19,0 %), three (n = 7, 4,6 %) or four (n = 8, 5,2 %) arthropod borne pathogens. In general the risk of infection was higher in dogs that were already infected with another pathogen. Altogether 28 (18.3 %) of the strays and 15 (9.8 %) of the dogs living in the animal shelter had a multiple infection. 75 of the 94 dogs infected with at least one of the tested pathogens were examined clinically. 33 (44.0 %) showed clinical symptoms. In 21 (67,7 %) of the 31 clinically examined dogs with multiple infection symptoms of disease were diagnosed. Twelve (27.3 %) of the 44 clinically examined dogs that were only infected with one pathogen had symptoms of some kind. Clinical symptoms varied considerably irrespective of the causative agent. The concordance of the IFAT and the ELISA for the diagnosis of Borrelia burgdorferi was 92.2 %. The serologic diagnosis of Borrelia burgdorferi with IFAT or ELISA can be regarded similarly suitable although ELISA seems to be somewhat more sensitive. In conclusion the risk of infection in Greece is particularly high for Ehrlichia canis, Borrelia burgdorferi and Leishmania infantum and Dirofilaria immitis as well. Babesia canis is obviously transmitted less. However dogs in poor condition or born in non endemic areas may acquire babesiosis in Greece. According to the results of this study Hepatozoon canis seems to be of negligible relevance in Greece. Due to the variability of the clinical symptoms and the frequent occurrence of multiple infections diagnosis based on the clinical picture only is not possible. Multiple infections should also be considered with respect to treatment. Infections with arthropod borne pathogens seem to occur less often in prophylactically treated dogs living under good conditions. Most of the examined ticks were identified as Rhipicephalus sanguineus (449 out of 457 specimens). Six nymphs and 419 adults were found. Of the adult Rhipicephalus sanguineus ticks 243 were males and 200 were females. Eight of the ticks were female Ixodes ricinus. In dogs originating from Greece as well as in dogs having accompanied tourists into Greece infestation with Rhipicephalus sanguineus is probable. As Rhipicephalus sanguineus is known to transmit Ehrlicha canis, Babesia canis vogeli and Hepatozoon canis prophylactic measures should include a suitable tick-control.

No description available.

No description available.

Over the last 20 years, Lyme disease has grown to become the most common vector-borne disease affecting Americans. Spread in the eastern U.S. primarily by the bite of Ixodes scapularis, the black-legged tick, the disease affects an estimated 329,000 Americans per year. Originally confined to New England, it has since spread across much of the east coast and has become endemic in Virginia. Since 2010 the state has averaged 1200 cases per year, with 200 annually in the New River Health District (NRHD), the location of our study. Efforts to geographically model Lyme disease primarily focus on landscape and climatic variables. The disease depends highly on the survival of the tick vector, and white-footed mouse, the primary reservoir. Both depend on the existence of forest-herbaceous edge-habitats, as well as warm summer temperatures, mild winter lows, and summer wetness. While many studies have investigated the effect of forest fragmentation on Lyme, none have made use of high-resolution land cover data to do so at the peridomestic level. To fill this knowledge gap, we made use of the Virginia Geographic Information Network’s 1-meter land cover dataset and identified forest-herbaceous edge-habitats for the NRHD. We then calculated the density of these edge-habitats at 100, 200 and 300-meter radii, representing the peridomestic environment. We also calculated the density of <2-hectare forest patches at the same distance thresholds. To avoid confounding from climatic variation, we also calculated mean summer temperatures, total summer rainfall, and number of consecutive days below freezing of the prior winters. Adding to these data, elevation, terrain shape index, slope, and aspect, and including lags on each of our climatic variables, we created environmental niche models of Lyme in the NRHD. We did so using both Boosted Regression Trees (BRT) and Maximum Entropy (MaxEnt) modeling, the two most common niche modeling algorithms in the field today. We found that Lyme is strongly associated with higher density of developed-herbaceous edges within 100-meters from the home. Forest patch density was also significant at both 100-meter and 300-meter levels. This supports the notion that the fine scale peridomestic environment is significant to Lyme outcomes, and must be considered even if one were to account for fragmentation at a wider scale, as well as variations in climate and terrain. / M.S. / Lyme disease is the most common vector-borne disease in the United States today. Infecting about 330,000 Americans per year, the disease continues to spread geographically. Originally found only in New England, the disease is now common in Virginia. The New River Health District, where we did our study, sees over 200 cases per year. Lyme disease is mostly spread by the bite of the black-legged tick. As such we can predict where Lyme cases might be found if we understand the environmental needs of these ticks. The ticks themselves depend on warm summer temperatures, mild winter lows, and summer wetness. But they are also affected by forest fragmentation which drives up the population of white-footed mice, the tick’s primary host. The mice are particularly fond of the interface between forests and open fields. These edge habitats provide food and cover for the mice, and in turn support a large population of ticks. Many existing studies have demonstrated this link, but all have done so across broad scales such as counties or census tracts. To our knowledge, no such studies have investigated forest fragmentation near the home of known Lyme cases. To fill this gap in our knowledge, we made use of high-resolution forest cover data to identify forest-field edge habitats and small isolated forest patches. We then calculated the total density of both within 100, 200 and 300 meters of the homes of known Lyme cases, and compared these to values from non-cases using statistical modeling. We also included winter and summer temperatures, rainfall, elevation, slope, aspect, and terrain shape. We found that a large amount of forest-field edges within 100 meters of a home increases the risk of Lyme disease to residents of that home. The same can be said for isolated forest patches. Even after accounting for all other variables, this effect was still significant. This information can be used by health departments to predict which neighborhoods may be most at risk for Lyme. They can then increase surveillance in those areas, warn local doctors, or send out educational materials.

Les réservoirs de Borrelia burgdorferi jouent un rôle clé pour assurer sa transmission dans l’environnement. Récemment, de nouveaux acaricides, comme le fluralaner, présentant une efficacité élevée contre les tiques et une longue durée d’action, ont été commercialisés sur le marché des médicaments vétérinaires. Cibler les réservoirs de B. burgdorferi, comme les souris du genre Peromyscus, avec le fluralaner permettrait : d’abord, d’en altérer le cycle endémique; ensuite, de réduire la densité de tiques infectées dans l’environnement ; et, ultimement, de réduire le risque de transmission de la maladie de Lyme (ML). En premier lieu, l’efficacité du fluralaner à tuer des larves de l’espèce Ixodes scapularis infestant des souris du genre Peromyscus a été démontrée en laboratoire. Lors d’infestations expérimentales, des doses de 12,5 et 50 mg/kg ont tué > 90 % des larves d’I. scapularis dans les 4 jours suivant l’administration du traitement. Cependant, cette efficacité a été réduite à moins de 5 % dans les 30 premiers jours. En second lieu, dans le cadre d’une étude controllée non randomisée, des appâts de fluralaner ont été administrés répétitivement à une population de micromammifères sauvages à des densités de 2,1 ou 4,4 appâts/1000 m2 pendant 3 à 4 ans. L’efficacité du traitement à réduire l’infestation des micromammifères a, au préalable, été évaluée. Les densités de 2,1 et 4,4 ont respectivement réduit de 68 % et 86 % le nombre de larves infestant les souris du genre Peromyscus. Seule la densité de 4,4 a réduit significativement le nombre de nymphes de 72 %. Après quoi, l’effet des deux densités d’appâts sur le cycle endémique de B. burgdorferi a été évalué par le biais de 3 paramètres : 1- la prévalence de Peromyscus spp. infectées (PIM), 2- la densité de nymphes en quête (DON), 3- la prévalence de nymphes infectées par B. burgdorferi (NIP). La prévalence d’infection des souris par B. burgdorferi a chuté de 60 à 37 % après deux années de déploiements du traitement. Le traitement a provoqué une réduction de la DON de 45 à 63 % dans les zones traitées. Aucun effet n’a été observé sur le dernier paramètre. En troisième lieu, la pharmacologie du fluralaner a été caractérisée chez des souris du genre Peromycus dans une étude réalisée en laboratoire. Les données recueillies ont été utilisées pour simuler et interpréter différents scénarios de traitement de souris Peromyscus spp. pour une durée équivalente aux pics d’activités des larves et des nymphes d’I. scapularis. Les simulations ont permis d’identifier un jeu de scénarios qui conféreraient une protection contre les tiques pendant toute la saison d’activité des stades immatures d’I. scapularis. L’étude de la pharmacologie des acaricides chez les espèces ciblées et la modélisation de la pharmacocinétique représentent des étapes cruciales pour développer ce type d’intervention. Cette thèse a permis d’entreprendre le développement d’une approche ciblant les réservoirs de B. burgdorferi en utilisant une molécule de la famille des isoxazolines. L’approche décrite a montré le potentiel d’altérer le cycle endémique de B. burgdorferi et de réduire la densité de nymphes infectées dans l’environnement. Plus généralement, en intégrant des études sur la pharmacologie du fluralaner en laboratoire et des études terrains, elle permet d’initier la réflexion quant à l’importance d’avoir un cadre de développement commun pour structurer la recherche autour de ce type d’approches. / Reservoirs of Borrelia burgdorferi play a key role in ensuring its transmission in the environment. Recently, new acaricides, such as fluralaner, with high efficacy against ticks and a long duration of action have been commercialized in the veterinary drug market. Targeting B. burgdorferi reservoirs, such as Peromyscus spp. mice, with fluralaner would yield the following benefits: first, alter its transmission in the environment; second, reduce the density of B. burgdorferi-infected ticks in the environment; and, ultimately, reduce the risk of Lyme disease (LD). Firstly, the efficacy of fluralaner for killing Ixodes scapularis larvae infesting Peromyscus mice was demonstrated in a laboratory setting. Doses of 12.5 and 50 mg/kg showed > 90% efficacy at killing I. scapularis larvae infesting Peromyscus spp. mice within 4 days of treatment during experimental infestations. However, efficacy was reduced to less than 5% beyond 30 days. Secondly, in a non-randomized controlled study, fluralaner baits were repeatedly administered to a wild population of small mammals at densities of 2.1 or 4.4 baits/1000m2 over a period of 3 to 4 years. First, the treatment's efficacy at reducing small mammal infestation was evaluated. Densities of 2.1 and 4.4 baits/1000m2 reduced the number of larvae infesting Peromyscus mice by 68% and 86% respectively. Only the density of 4.4 baits/1000m2 significantly reduced the number of nymphs by 72%. Next, the effect of the both bait densities on the B. burgdorferi endemic cycle was evaluated through 3 parameters: i- the prevalence of B. burgdorferi-infected Peromyscus spp. mice (PIM), ii- the density of questing nymphs (DON), and iii- the prevalence of B. burgdorferi in questing nymphs (NIP). Mouse infection by B. burgdorferi decreased from 60 to 37% following two years of treatment deployment. The treatment reduced the DON by 45-63% in the treated areas but no effect was observed on B. burgdorferi prevalence in the NIP. Thirdly, fluralaner pharmacology was characterized in Peromycus mice in a laboratory setting. The data collected were used to simulate and interpret different treatment scenarios for Peromyscus mice over a period corresponding to the peak activities of I. scapularis larvae and nymphs. The simulations identified a set of treatment scenarios that confer protection against ticks for an entire season of I. scapularis activity. The results indicate that investigation of acaricide pharmacology in target species and pharmacokinetic modeling are critical steps in the development of this type of intervention. This thesis initiated the development of an approach targeting B. burgdorferi reservoirs using a molecule of the isoxazoline family. This approach showed potential for disrupting the endemic cycle of B. burgdorferi and reducing the density of B. burgdorferi-infected nymphs in the environment. More broadly, by integrating studies on fluralaner pharmacology in laboratory and field settings, this work has initiated a reflection on the importance of establishing a general development framework to structure research around this type of approach.

Indiana University-Purdue University Indianapolis (IUPUI) / The Lyme disease agent, Borrelia burgdorferi, has a complex system that allows it to thrive in the harsh and distinct environments of its tick vector and mammalian host. Although it has been known for some time that the Borrelia oxidative stress regulator protein (BosR) plays a necessary role in mammalian infectivity and functions as a transcriptional regulator of alternative sigma factor RpoS, very little is known about its mechanism of action, other than the suggestion that BosR activates rpoS transcription by binding to certain upstream regions of the gene. In our studies, we performed protein degradation assays and luciferase reporter assays for further understanding of BosR function. Our preliminary findings suggest that BosR is post-transcriptionally regulated by an unknown protease and may not need to bind to any rpoS upstream regions in order to activate transcription. We also describe the construction of luciferase reporter systems that will shed light on BosR’s mechanism of action. We postulate the provocative possibility that unlike its homologs Fur and PerR in other bacterial systems, BosR may not utilize a DNA-binding mechanism in order to fulfill its role as a transcriptional regulator to modulate virulence gene expression.

Submitted by Cleide Dantas (cleidedantas@ufpa.br) on 2014-07-28T14:14:26Z No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_FrequenciaAnticorposAntiBorrelia.pdf: 513114 bytes, checksum: c4c606deec6650eaae49134f3922e92a (MD5) / Approved for entry into archive by Ana Rosa Silva (arosa@ufpa.br) on 2014-09-11T15:33:58Z (GMT) No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_FrequenciaAnticorposAntiBorrelia.pdf: 513114 bytes, checksum: c4c606deec6650eaae49134f3922e92a (MD5) / Made available in DSpace on 2014-09-11T15:33:58Z (GMT). No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_FrequenciaAnticorposAntiBorrelia.pdf: 513114 bytes, checksum: c4c606deec6650eaae49134f3922e92a (MD5) Previous issue date: 2006 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A Borreliose de Lyme é uma doença multissistêmica causada pela espiroqueta Borrelia burgdorferi latu sensu e transmitida por carrapatos ixodideos, acometendo seres humanos e animais domésticos, tendo como reservatórios naturais os animais silvestres, sendo ainda considerada uma zoonose de ampla distribuição geográfica. Foram coletadas 300 amostras sanguíneas de eqüinos aparentemente sadios, procedentes dos municípios de Ananideua, Belém, Benevides, Castanhal, Marituba e Santa Izabel do Pará da mesorregião metropolitana de Belém – Pará. O sangue foi coletado pela veia jugular e os soros foram analisados através do ensaio de imunoadsorção enzimática (ELISA) indireto no Laboratório de Doenças Parasitárias na Universidade Federal Rural do Rio de Janeiro. O objetivo do trabalho foi verificar a soroepidemiologia da Borrelia burgdorferi na mesorregião metropolitana de Belém. A freqüência de soropositividade foi de 26,67% (n=80), sendo 72 (24%) com título de 1:800, seis (2%) a 1:1600 e dois (0,6%) a 1:3200. A soropositividade entre os sexos foram valores aproximados sendo 13,67% nas fêmeas e 13% nos machos. A freqüência entre os animais de raças e mestiços foram 9% e 18%, respectivamente. Não houve diferença significativa de animais soropositivos quanto aos municípios, sexo, raça e faixa etária. A freqüência encontrada corrobora a hipótese da ocorrência de Borrelia sp. na região estudada. / Lyme borreliosis is a multisistemic disease caused by the spirochete Borrelia burgdorferi latu sensu and transmited by ixodide ticks, affecting both humans beings and domesticated animals and having wild animals as Its natural reservoirs, besides of being a zoonosis of wide geographic distribution. We picked 300 blood samples of apparently health horses from the cities of Ananideua, Belém, Benevides, Castanhal, Marituba and Santa Izabel of the Pará, all included in the metropolitan mesoregion of Belém – Pará. The samples were picked up by the jugular vein and the serum analised through indirect Enzyme-Linked Immunosorbent Assay (ELISA) in the Laboratory of Parasitic Diseases of the Federal Rural University of Rio de Janeiro. The aim of this work was to verify the serum epidemiology of Borrelia burgdorferi in the metropolitan mesorregion of Belém. The frequency of serum positivity was 26,67% (n=80), being 72 (24%) with title of 1:800, six (2%) with 1:1600 and two (0,6%) with 1:3200. The serum frequency by gender was approximately 13,67% of females and 13% of males. The thoroughbred and the half-breed animals had 9% and 18% of frequency, respectively. There was no significative difference in the serum positivity with regard to cities, gender, race and age. The frequency found corroborates the hypothesis of occurrence of Borrelia sp. in the studied region.

No description available.