Podcasts about Anaplasma

In This Episode: Nicole Bell, CEO of Galaxy Diagnostics, shares her journey from personal experience with Lyme disease in her family to leading innovation in diagnostics. The connection between tick-borne pathogens and chronic diseases like early-onset Alzheimer's. Why traditional Lyme disease tests often fail and the need for direct detection methods. The latest advancements in testing for Borrelia, Bartonella, Babesia, and other stealth pathogens. The impact of Nicole's "State of Lyme Disease Research" report on the Lyme community. What patients and healthcare providers need to know about cutting-edge diagnostic tools. Why This Matters: An increasing number of studies link flea- and tick-borne pathogens to chronic illnesses affecting the joints, heart, and central nervous system. These infections mimic conditions like lupus and fibromyalgia, making them hard to diagnose without advanced testing. Galaxy Diagnostics offers specialized tests to detect Lyme Borrelia, Bartonella, Babesia, Anaplasma, Ehrlichia, and Rickettsia species—providing a much-needed solution for patients seeking accurate diagnoses. Resources & Links:

Chronic Lyme disease, characterized by persistent symptoms that can linger for months or even years, is a growing concern for both patients and healthcare providers. Traditionally, Lyme disease has been seen as a straightforward infection—diagnosed early, treated with antibiotics, and resolved. However, for a significant number of people, Lyme disease can evolve into a chronic condition, marked by ongoing fatigue, pain, and cognitive issues. But what if much of this chronic suffering could be prevented? Recent insights suggest that the progression to chronic Lyme disease might be mitigated, or even prevented, by focusing on several key factors. This approach is similar to how we prevent other chronic diseases like type 2 diabetes or heart disease: through early detection and appropriate treatment. **The Importance of Early Diagnosis and Treatment** One of the most critical steps in preventing chronic Lyme disease is the early diagnosis and treatment of the infection. When a patient presents with a tick bite or early symptoms of Lyme disease, such as the characteristic erythema migrans rash, it's vital to begin treatment promptly. Early intervention with antibiotics like doxycycline can significantly reduce the risk of the infection progressing to a chronic stage. Moreover, it's essential to maintain a vigilant follow-up process. Even after initial treatment, patients should be asked to return for evaluation if symptoms persist or recur. Lyme disease can sometimes be insidious, with symptoms reappearing after what seems like successful treatment. Regular follow-ups allow healthcare providers to monitor the patient's progress and catch any signs of relapse early. **Look for Co-Infections** Co-infections are another critical factor in the progression of Lyme disease to a chronic condition. Ticks often carry more than just Borrelia burgdorferi, the bacteria responsible for Lyme disease—they can also transmit other pathogens like Babesia, Bartonella, and Anaplasma. These co-infections can complicate the clinical picture and may require different treatments. It's important to test for and consider co-infections, especially if a patient's symptoms do not resolve with standard Lyme treatment. Addressing all possible infections early on can prevent the persistence and worsening of symptoms. **Consider Retreatment** In cases where initial treatment does not fully resolve the symptoms, it may be necessary to consider retreatment. Clinical judgment plays a crucial role here. While some cases of Lyme disease respond well to the standard course of antibiotics, others may require a more prolonged or alternative treatment approach. Retreatment should be considered for patients who continue to experience significant symptoms, particularly if they show signs of ongoing infection or co-infections. **Follow-Up Care is Crucial** Consistent follow-up care is essential in managing Lyme disease and preventing its progression to a chronic state. Even after symptoms appear to have resolved, patients should be monitored for any signs of recurrence or new symptoms. Chronic Lyme disease can sometimes develop months or even years after the initial infection, making long-term follow-up a key component of care. **Environmental Awareness and Preventive Measures**  **Raising Awareness and Improving Access to Care** 

Introduction to Dr. Omar Morales Welcome to this episode where we delve into the world of Lyme disease with our esteemed guest, Dr. Omar Morales, founder of the Lyme Mexico Clinic, renowned researcher, and dedicated member of the Board of Directors of ILADS (International Lyme and Associated Diseases Society). Join us as Dr. Morales shares his journey from construction to medicine, his pivotal role in Lyme disease treatment, and his advocacy efforts with ILADS. Dr. Morales's Journey: From Construction to Medicine Unexpected Beginnings: Hear how a promise of a truck from his father led Dr. Morales to a career in medicine. Family Influence and Medical Career: Learn about the familial influences that shaped Dr. Morales's path and his early medical experiences. Establishing the Lyme Mexico Clinic Clinic Origins: Discover the story behind the founding of the Lyme Mexico Clinic and its evolution into a leading Lyme disease treatment center. Patient-Centered Care: Dr. Morales discusses his approach to individualized patient care and the clinic's commitment to understanding and treating Lyme disease. Lyme Disease: A Complex Challenge Defining Lyme: Delve into the complexities of defining Lyme disease, chronic vs. acute presentations, and the systemic nature of the illness. Insights from ILADS: As a member of the ILADS Board of Directors, Dr. Morales shares his perspectives on the latest Lyme disease research and treatment approaches. The Detective Work in Understanding Lyme Disease: Dr. Morales emphasizes the complexity of Lyme disease, noting the existence of five different subtypes of Lyme and about 300 different sub-strains of Borrelia. He also highlights the significance of co-infections like Bartonella, Babesia, and Anaplasma, as well as the importance of considering both the genetics of the pathogen and the genetics of the patient in treatment strategies. Pathogen-Induced Immune Responses: Dr. Morales has found that Bartonella can trigger an overactive immune system or autoimmunity in patients, whereas Borrelia (Lyme) more often triggers immune suppression, especially in those with genetic predispositions. This distinction is critical for understanding why some chronic Lyme patients have barely functioning immune systems while others have overactive immune systems. Innovative Approaches to Lyme Treatment Treatment Philosophy: Explore Dr. Morales's "less is more" approach to Lyme treatment, emphasizing the importance of timely and effective interventions. Advancements in Treatment: Learn about the innovative treatment strategies employed at the Lyme Mexico Clinic, including intravenous treatments and the clinic's stance on maintaining patient homeostasis. Four-Part Treatment Framework: Dr. Morales outlines his comprehensive treatment framework, focusing on treating infections, targeting persistent forms of bacteria like biofilm, supporting the immune system, and detoxing. He provides specific treatments for each of these areas during the interview. Neurological Symptoms of Lyme Disease: Research Insights In-Depth Discussion: We dive deep into the neurological symptoms associated with Lyme disease, an area where Dr. Morales has conducted extensive research. 2024 Fall ILADS Conference in Germany: Dr. Morales will be presenting his significant findings on the neurological implications of Lyme disease at the upcoming ILADS Conference, showcasing his contributions to understanding and treating this complex aspect of Lyme. Research and Collaboration in Lyme Disease The Role of Research: Understand the critical role of ongoing research in advancing Lyme disease treatment and the importance of collaboration within the medical community. Advocacy for Lyme Disease Awareness: Dr. Morales discusses his advocacy work with ILADS and the importance of raising awareness and support for Lyme disease research. Patient Advocacy and Care Strategies Commitment to Patients: Hear about Dr. Morales's dedication to patient care, including offering free initial consultations and personalized treatment plans. Understanding Lyme Disease: Gain insights into the detective work involved in diagnosing and treating Lyme disease and the importance of listening to and understanding patients' experiences. Conclusion: A Future Free from Lyme We wrap up our Tick Boot Camp Podcast conversation with Dr. Morales by discussing the future of Lyme disease treatment and the importance of community support, early detection, and comprehensive care. Dr. Morales's work with the Lyme Mexico Clinic and ILADS exemplifies the ongoing efforts to improve lives affected by Lyme disease.

Preventative diagnostics is a critically important component of veterinary care. In the U.S., only 40-50% of pet owners take their pets to the vet once a year. While the costs of vet visits continue to increase, the cost of not taking your pet to the vet regularly can far outweigh the cost of a vet visit. With the cost of a simple vet visit now approaching $100 in some areas, and the costs of any tests and treatments further adding to the total cost of the bill, many pet parents take the approach of not bringing their pet to the vet unless they see overt signs that their pet is struggling. However, dogs and cats are very good at masking issues that they might be having and not outwardly showing any signs of pain, and without regular checkups, issues that could have been easily managed if they were caught early can quickly turn into much more catastrophic health events. On the latest episode of Petworking, I was joined by Dr. Ben Hantler, DVM, MBA, and Senior Product Manager at Heska, and my colleague, Jessica Stewart, Senior Manager at Maia Strategy Group. Prior to joining Maia, Jessica was a Diagnostic Support Consultant at IDEXX. In the course of our conversation, Ben noted that 20-30% of serious cases could have been mitigated or prevented with regular veterinary visits. While pet insurance is undoubtedly a key driver in getting pet parents to visit the vet more frequently, Ben, Jess, and I kept our conversation focused on diagnostics. We touched on the variety of tests available, from organ function assessments and blood counts, to infectious disease screenings like heartworm testing. The necessity of these getting these tests on an annual basis cannot be understated, when it comes to catching potential health issues early. When we adopted Peony, we were told she was heartworm-negative. However, a month after we got her, I brought her to the vet, because she seemed to be experiencing pain in her hind legs, and our trainer suspected she might have pulled a muscle. Wanting to eliminate tick-borne illnesses as a potential cause of her symptoms, our vet ran a SNAP test. While it was negative for Lyme, Ehrlichia, and Anaplasma, it did show that she was positive for heartworm. Had we not gotten Peony tested, it's very likely that we would have caught the infection too late to treat her. The American Heartworm Society notes that even the most sensitive heartworm test can fail to detect the presence of antigen when levels of circulating antigen are low due to the presence of immature worms. Our vet told us that this was likely how we were told Peony was heartworm-negative when we adopted her. Unfortunately, if left untreated, heartworm can cause severe organ damage, heart failure, and respiratory issues, and can have fatal consequences. Even catching it early, it was a several-month process to treat Peony, during which we had to restrict her activity to two 5-minute walks per day. In addition to catching potentially fatal diseases like heartworm, regular diagnostic testing can provide pet owners with a baseline that they can refer to as their pet ages. Having year-over-year data on biomarkers like blood levels, kidney function, etc. can alert your vet to changes that may indicate potential health issues in your pet, even in the absence of symptoms. As part of this discussion, Ben, Jess and I delved into the potential use of biometric data from pet trackers, and the need to expand preventative diagnostic testing to a direct-to-consumer model. There is significant potential to enhance the quality of care through the provision of consumer-collected data to veterinarians, but a limiting factor that needs to be addressed is ensuring consistent quality in at-home sample collection. Ultimately, this is a field at the cusp of significant evolution, promising a future where preventative care, supported by technological advancements, could drastically improve the lives of pets and pet owners alike.

AABP Executive Director Dr. Fred Gingrich is joined by past AABP Board of Directors member from District 3, Dr. Lee Jones. Jones is an associate professor at the University of Georgia in the Beef Production Medicine department. Jones was the first author on the paper titled “Seroprevalence and molecular detection of Anaplasma marginale-infected beef herds in Georgia, USA”, which was published in The Bovine Practitioner Volume 56 Number 2 (2022). We review the transmission, clinical signs, and diagnostic tests for bovine anaplasmosis. The objective of the study was to conduct and compare seroprevalence and molecular detection of Anaplasma marginale-infected beef herds in Georgia and to identify herd risk factors associated with A. marginale-positive herds. Blood samples were collected from 1,059 adult beef cattle (≥ 2 years) from 33 herds. Overall, 8.12% of cattle and 42% of herds were cELISA antibody-positive. Seventy-seven percent of plasma samples from a subset of corresponding seropositive samples (n = 73) were PCR-positive. Jones reviews the risk factors associated with bovine anaplasmosis and how veterinarians can become involved in prevention and control programs for their clients. Education is important because 27% of the survey respondents were not aware of the disease even with a high percentage of herds positive. We also discuss the importance of ensuring appropriate delivery of VFD medication for control of anaplasmosis as part of a control program. Relevant publication:Jones, A. L., Berghaus, R. D., Kalatari, A. A., Credille, B., Naikare, H. K., Heins, B., Saliki, J., & Wilkes, R. P. (2022). Seroprevalence and molecular detection of Anaplasma marginale infected beef herds in Georgia, USA. The Bovine Practitioner, 56(2), 70–78. https://doi.org/10.21423/bovine-vol56no2p70-78  

Join Yvonne Brandenburg, RVT, VTS SAIM and Jordan Porter RVT, LVT, VTS SAIM as we talk about: The big four!!! Tick borne bacterial diseases; anaplasmosis, ehrlichia, lyme, and rocky mountain spotted fever, OH MY! Ever wonder how ticks transmit these diseases? Well you're about to find out, prepare to get squiggy.    Question of the Week Have you had a personal experience with one of these tick borne illnesses, like human experience.  Leave a comment at https://imfpp.org/episode89   Resources We Mentioned in the Show  Canine Rocky Mountain Spotted Fever. Parasitology Compendium. July 2005 (Vol 27, No 7). Rochelle M. Low , DVM , Jennifer L. Holm , DVM , DACVECC  https://www.vetfolio.com/learn/article/canine-rocky-mountain-spotted-fever Rocky Mountain Spotted Fever in Dogs. (Rickettsia rickettsii Infection). Janet E. Foley , DVM, PhD, University of California, Davis. https://www.merckvetmanual.com/generalized-conditions/rickettsial-diseases/rocky-mountain-spotted-fever-in-dogs    Anaplasmosis. Becky Lundgren, DVM. Date Published: 04/08/2014. https://veterinarypartner.vin.com/default.aspx?pid=19239&id=6191808 Merrill, L. (2012). Small Animal Internal Medicine for Veterinary Technicians and Nurses. Ames: Wiley - Blackwell.   Thanks so much for tuning in. Join us again next week for another episode!  Want to earn some RACE approved CE credits for listening to the podcast? You can earn between 0.5-1.0  hour of RACE approved CE credit for each podcast episode you listen to.    Join the Internal Medicine For Vet Techs Membership to earn and keep track of your continuing education hours as you get your learn on!   Join now! http://internalmedicineforvettechsmembership.com/   Get Access to the Membership Site for your RACE approved CE certificates Sign up at https://internalmedicineforvettechsmembership.com  Get Access to the Technician Treasure Trove  Sign up at https://imfpp.org/treasuretrove    Thanks for listening!  – Yvonne and Jordan 

Why You Should Listen: In this episode, you will learn about healing from mold and Lyme disease. About My Guests: My guest for this episode is Dr. Diane Mueller. Diane Mueller, ND, DAOM, LAc is a survivor of IBS, Lyme disease, and mold illness. Dr. Diane's journey to heal herself led her to complete two doctorate degrees in holistic health care. She has a Doctorate degree in Naturopathic Medicine as well as a Doctorate degree in Acupuncture and Oriental Medicine. She is passionate about bringing research, understanding, and compassion to those with these conditions. She has co-authored the book "Use Your Mind to Heal Your Mold and Lyme: A Survivor's Guide". Her practice, the Medicine with Heart Clinic, treats people from around the country. She co-owns an online functional medicine school, the Medicine with Heart Institute, where she trains clinicians around the world in functional medicine. Her recent book “Use Your Mind to Heal Your Mold and Lyme: A Survivor's Guide” shares many of the strategies that she used to recover her own health and the health of many of her patients. Key Takeaways: - Should mold be treated before Lyme disease? - Is killing the bug important for recovering health? - How can pulsing antimicrobials be part of a protocol? - What are persisters, and how are they addressed? - Can mycotoxins lead to leaky gut? - Does fungal colonization occur after exposure to water-damaged buildings? - How does Bartonella impact the lymphatics? - How do Ehrlichia and Anaplasma negatively impact our mitochondria? - What is the role of viruses and retroviruses in chronic illness? - Can medicine mushrooms be used in those with mold illness or fungal overgrowth? - What is the role of bile transporters in detoxification? - How might manganese or hyaluronic acid be used as "feeders"? - How might pulsing be used to avoid sensitization to therapeutic interventions? - What properties does cistus have that make it a useful tool? - Why is it important to approach biofilm treatment with caution? - What is the role of autophagy in cleansing? - What can negative thoughts do to the physical body? - Where can patients find support? Connect With My Guest: http://MedicineWithHeart.com Related Resources: Book - Use Your Mind to Heal Your Mold and Lyme: A Survivor's Guide Body, Mind, Mold and Lyme Summit Interview Date: April 6, 2021 Transcript: To review a transcript of this show, visit http://BetterHealthGuy.com/Episode143 Additional Information: To learn more, visit http://BetterHealthGuy.com. Disclosure: BetterHealthGuy.com is an affiliate of Amazon.com and HeySummit.com Disclaimer: The content of this show is for informational purposes only and is not intended to diagnose, treat, or cure any illness or medical condition. Nothing in today's discussion is meant to serve as medical advice or as information to facilitate self-treatment. As always, please discuss any potential health-related decisions with your own personal medical authority.

Raul Mascarenhas explica o que é a Tristeza Parasitária bovina, como agem os micro-organismos envolvidos (Babesia e Anaplasma) e seus sinais clínicos (febre, anemia e mudanças no comportamento). #tristezaparasitaria #babesiose Seja um apoiador! Nosso PIX para caso você queira ser um apoiador é o e-mail casaagropecuaria@raulmascarenhas.com Acesse mais conteúdos gratuitos em: https://www.facebook.com/casa.agropecuaria.raulmascarenhas https://raulmascarenhas.com/ Twitter @CasaMascarenhas --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app --- Send in a voice message: https://anchor.fm/raul-mascarenhas/message Support this podcast: https://anchor.fm/raul-mascarenhas/support

Here we learn about a disease that is deadly to cattle called Anaplasma. And maybe more about Dr. Nathan's Game of Thrones obsession that you wanted.

 Don talks with Kindred Spirits Veterinary Clinic’s Dr. Mark Hanks about infectious diseases and parasites that affect our pets. We discuss Canine Lyme Disease and Anaplasma and other tick-borne diseases including diagnosis, treatment, and prevention. We also discuss parasites, and Dr. Hanks recommended annual fecal testing for pets. Tick-borne diseases have become a serious health issue for pets in people and Maine. If you want to learn more, you will not want to miss this show. You can hear The Woof Meow Show on Z62 Retro Radio, AM620, and WKIT HD3 at 9 AM on Saturday. If you are not near a radio, listen on your computer at http://bit.ly/AM620-WZON or your smartphone or tablet with the free WZON 620 AM app. A podcast of the show is typically posted immediately after the show. You can download this show and others at http://woofmeowshow.libsyn.com/, at Don’s blog http://bit.ly/Words-Woofs-Meows and the Apple iTunes store.

Learn about the study that demonstrated that Simparica blocked the transmission of Lyme and Anaplasmosis in treated dogs. Listeners will learn about a published, peer-reviewed study designed to test tick protection by Simparica. In this study, Simparica- and placebo-treated dogs were infested with Ixodes scapularis (aka deer) ticks on day 21 and day 28 after treatment. These ticks were infected with Borrelia burgdorferi and Anaplasma phagocytophilum which was subsequently transmitted to control dogs, as evidenced by positive laboratory tests. Simparica-treated dogs, on the other hand, tested negative at all time points and with every test methodology. This demonstrates that Simparica kills tick quickly, and blocked transmission of these pathogens in these dogs. This quick and easy download is designed to help you stay up to date on the latest advances in veterinary medicine. Listen to this podcast at your convenience — at home, on your commute, during your lunch break…anytime!

We are joined by Dr. Memo Arroyo, medicine clinician at the Ontario Veterinary College, who gives an update on the OAHN Equine Project: "Seroprevalence of Borrelia burgdorferi and Anaplasma phagocytophilum infection in Ontario horses." Dr. Arroyo also discusses the upcoming OVC Equine Research Update night on December 15th. 

Kleinsäuger sind essentiell für die Entwicklung und die Verbreitung von subadulten Schildzecken. Den Kleinsäugern kommt so eine wichtige Rolle als potentielle Reservoirwirte für Zecken-übertragene Pathogene zu. Die Ziele dieser Studie waren unterschiedliche Zecken-übertragene Pathogene in wildlebenden Kleinsäugern nachzuweisen und die Reservoirfunktion der jeweiligen Kleinsäugerarten, im Zusammenhang mit unterschiedlich strukturierten Habitaten, zu evaluieren. Zwischen 2012 und 2013 wurden Kleinsäuger an drei unterschiedlich strukturierten Standorten gefangen: (1) an einem Stadtpark in Regensburg, (2) an einem silvatischen Standort in Tussenhausen im Unterallgäu und (3) an einem renaturierten Standort, der in der Nähe von Leipzig in Sachsen liegt. Zusätzlich wurden Zecken im Jahr 2013 am Waldstandort geflaggt. DNA wurde aus Blut-, Milz- und Gonaden-Proben der Mäuse und aus Mäuseneonaten extrahiert. Auf den Mäusen befindliche Zecken wurden abgesammelt. Aus diesen und den wirtssuchenden Zecken wurde ebenfalls DNA extrahiert. Zusätzlich wurden bereits vorhandene DNA-Proben aus wirtssuchenden Zecken aus den Jahren 2009-2013 bzw. 2011-2012 vom urbanen bzw. vom silvatischen Standort untersucht. Die Proben wurden mittels konventioneller oder Real-Time PCR auf Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis (CNM) und Babesia microti untersucht. Insgesamt wurden 631 Kleinsäuger zehn verschiedener Arten gefangen (4 Apodemus agrarius, 7 Microtus arvalis, 1 M. agrestis, 396 Myodes glareolus, 2 Mustela nivalis, 5 Sorex coronatus, 1 Sorex araneus, 1 Talpa europaea, 36 Ap. sylvaticus, 178 Ap. flavicollis). Davon wurden insgesamt 36 Mäuse im Stadtpark, 243 am silvatischen und 352 am renaturierten Standort, wo die größte Artenvielfalt vorherrschte (n=8), gefangen. Insgesamt wurden 3.391 Zecken drei verschiedener Arten (8 Ixodes trianguliceps, 3.250 Ixodes ricinus, 133 Dermacentor reticulatus) abgesammelt. CNM wurde in insgesamt 28,6 % der Kleinsäuger nachgewiesen. Dabei waren 31,6 % My. glareolus, 28,1 % Ap. flavicollis, 57,1 % M. arvalis und 2,7% Ap. sylvaticus positiv. Die Prävalenzen unterschieden sich signifikant beim Vergleich der jeweiligen Standorte, wobei die Infektionsrate am renaturierten Standort am höchsten war (χ²: 13,4; p: 0,0004). Insgesamt waren 3,8 % der gesogenen und 2,2 % der wirtssuchenden Zecken positiv. In den untersuchten Kleinsäugerföten bzw. -Neonaten, die von positiven Muttertieren stammten, war die Prävalenz für CNM 31,8 %. Insgesamt 60,0 % der positiven Muttertiere hatten wenigstens einen positiven Foetus oder Neonaten. Anaplasma phagocytophilum wurde zu einem geringen Prozentsatz in Nagern festgestellt (0,0-5,6 %), wobei es keinen signifikanten Unterschied zwischen den Standorten, Jahren und Kleinsäugerarten gab. Jedoch waren gesogene Nymphen (I. ricinus) signifikant häufiger befallen als gesogene Larven (χ²: 25,1; p:

Anaplasma phagocytophilum (Ap) is a gram-negative, obligate intracellular bacterium that is able to infect different animal species and humans worldwide. Based on DNA sequencing, Ap has newly been reallocated from the genus Ehrlichia to the genus Anaplasma in the family Anaplasmataceae (DUMLER et al. 2001). In humans and animals, the clinical signs of Ap infection vary from mild symptoms to severe clinical outcomes, including death. However, the disease generally presents as undifferentiated fever accompanied by leucopenia, thrombocytopenia and increased serum transaminase activities (DUMLER et al. 2005; DUMLER et al. 2007; RIKIHISA 2011). Hard-bodied ticks of the genus Ixodes (family Ixodidae) are the main vectors for Ap dissemination. Compared to other pathogens such as Neorickettsia and Wolbachia spp., which can be transmitted from adult ticks to their offspring, Anaplasma and Ehrlichia spp. are the only Rickettsiales that are not transmitted transovarially (RIKIHISA 2011). Thus, ticks need to acquire Ap through blood feeding from infected hosts to complete the life cycle of Ap. During attachment of the tick, the bacterium is released by salivary secretion and is transmitted to the host. It is known that Ap multiplies within membrane-bound vacuoles (or called ‘morulae’) in the cytoplasm of peripheral granulocytes. The binding and infection of bacteria depends on the tetrasaccharide sialyl Lewisx (sLex or CD15s) of P-selectin glycoprotein ligand 1 (PSGL-1) on the surface of host cells, a factor expressed on peripheral granulocytes and HL-60 cells (GOODMAN et al. 1999; HERRON et al. 2000; RENEER et al. 2006; RENEER et al. 2008). Only little information is known about the transmission pathway of Ap after tick bite in the very early stage of infection. It is described that Ap is able to evade and replicate within microvascular endothelial cells in vitro (MUNDERLOH et al. 2004), while endothelial cells lining the inner lumen of blood vessels allow them to easily interact with any circulating blood cells. Since granulocytes do not return back to the blood stream after extravasation, it is reasonable to postulate that Ap evades and replicates within microvascular endothelial cells in the initial transmission, and subsequently transmits into peripheral granulocytes for ongoing dissemination. Therefore, the objective of the study was to establish a flow culture model that mimics the physiological environment in the blood vessel to study the possible transmission pathway of Ap between endothelial cells and polymorphonuclear leukocytes (PMNs). For this purpose, a novel ex vivo flow culture system was established. For experimental setup, human microvascular endothelial cell line (HMEC-1) and primary human dermal microvascular endothelial cells (HDMEC) were used. Under static conditions, Ap evades endothelial cells within 24 h, supporting the hypotheses that endothelial cells might be the first infection site of the pathogen in the host. Thereby a high level of interleukin-8, a chemokine that is known to recruit PMNs, secreted by Ap-infected endothelial cells was detected. Using the investigated flow culture model, it was shown for the first time, that Ap is able to translocate from endothelial cells to PMNs under dynamic flow conditions. Furthermore, under defined shear stress, an increased binding of PMNs to Ap-infected endothelial cells monolayer was observed, resulting from the elevated expression of adhesion molecules associated with PMNs recruitment on endothelial cells. The flow culture model investigated in this study can be used to study the interaction between Ap-infected endothelial cells and PMNs under physiological flow conditions, and is therefore helpful to study the infection mechanism in the early stage of Ap dissemination in the host.

The study aimed to characterize the occurrence of tick-borne diseases within a herd in an endemic region in Brazil, focusing on two different aspects: (a) to determine the occurrence of the three pathogens Anaplasma marginale, Babesia bovis and Babesia bigemina using direct and indirect detection methods and (b) to analyze the incidence of genetic diversity among A. marginale isolates based on a Major Surface Protein (MSP) as a marker of variation within the farm.

Sat, 9 Feb 2013 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/16638/ https://edoc.ub.uni-muenchen.de/16638/1/Barth_Charlotte.pdf Barth, Charlotte

Background: The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods: Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results: 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion: Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

Background: Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010) and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. Results: The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae) in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae) in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae) in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25), B. divergens (n = 1), B. divergens/capreoli (n = 1), B. gibsoni-like (n = 1), R. helvetica (n = 272), R. monacensis IrR/Munich (n = 12) and unspecified R. monacensis (n = 1). The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27), but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. Conclusions: I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green areas are likely to remain in the research focus on public health issues.

Background: Equine Granulocytic Anaplasmosis (EGA) is caused by Anaplasma phagocytophilum, a tick-transmitted, obligate intracellular bacterium. In Europe, it is transmitted by Ixodes ricinus. A large number of genetic variants of A. phagocytophilum circulate in nature and have been found in ticks and different animals. Attempts have been made to assign certain genetic variants to certain host species or pathologies, but have not been successful so far. The purpose of this study was to investigate the causing agent A. phagocytophilum of 14 cases of EGA in naturally infected horses with molecular methods on the basis of 4 partial genes (16S rRNA, groEL, msp2, and msp4). Results: All DNA extracts of EDTA-blood samples of the horses gave bands of the correct nucleotide size in all four genotyping PCRs. Sequence analysis revealed 4 different variants in the partial 16S rRNA, groEL gene and msp2 genes, and 3 in the msp4 gene. One 16S rRNA gene variant involved in 11 of the 14 cases was identical to the "prototype" variant causing disease in humans in the amplified part [GenBank: U02521]. Phylogenetic analysis revealed as expected for the groEL gene that sequences from horses clustered separately from roe deer. Sequences of the partial msp2 gene from this study formed a separate cluster from ruminant variants in Europe and from all US variants. Conclusions: The results show that more than one variant of A. phagocytophilum seems to be involved in EGA in Germany. The comparative genetic analysis of the variants involved points towards different natural cycles in the epidemiology of A. phagocytophilum, possibly involving different reservoir hosts or host adaptation, rather than a strict species separation.

The major aim of this thesis was to identify novel genes of T. uilenbergi through establishment and screening of a merozoite cDNA library with the eventual goal to develop diagnostic tools using identified genes for detection of Theileria infections. The experiments were initiated by infection of sheep using T. uilenbergi stock. When parasiteamia rose, blood was collected and the merozoites were purified. Messenger RNA was isolated from purified merozoites was then utilized to establish a cDNA library. The library was titrated to be 6 x 108 pfu/ml and the recombinant clones were estimated to be 70%. Random PCR identification of the library indicated all of the inserts were of parasite origin, indicating the usefulness of the library for the identification of new genes. Random PCR amplification of inserts of the cDNA library led to the discovery of 12 single clones, among which Clone 2, 9 and 26 exhibited a high degree of identity, especially at the 3' terminus and 3' untranslated region, indicating that they belong to the same gene family. Furthermore, PCR designed to target Clone 2 amplified again four variant genes from genomic DNA of T. uilenbergi and one from genomic DNA of T. luwenshuni, suggesting this gene family is likely isolate-specific since the DNA samples for PCR were not derived from the same parasite isolate used for library construction. Sequence analysis of another genomic fragment generated with primers targeting the 3' untranslated region of the Clone 26 sequence showed that both 5' and 3' termini were highly identical to the Clone 2 gene family and these homologous termini were separated by a 136 bp sequence fragment highly identical to the 3' untranslated region of the Clone 2 gene family, indicating Clone 2 gene family members are tandemly arranged. Bioinformatic analysis of cDNA sequences of the Clone 2 gene family indicated these genes contain signal peptides and encode potential immunogenic proteins. Analysis of recombinantly expressed Clone 2 revealed immunoreactivity with sera from Theileria-infected animals from China. No cross reaction with sera of T. lestoquardi-, Babesia motasi- or Anaplasma ovis- infected animals was observed, indicating a potential specificity of this gene family. The features of the Clone 2 gene family are similar to the Tpr gene family of T. parva, which is believed to play a role in concerted evolution. Based on the highly conserved region of the Clone 2 gene family, a set of six primers were designed for the development of a loop mediated isothermal amplification (LAMP). The established assay allowed the detection of T. uilenbergi and T. luwenshuni infections simultaneously and the reaction could be simply accomplished by incubation at 63ºC for 15 min. The specificity of the reaction was confirmed through EcoRI restriction enzyme digestion analysis and sequencing. The assay was sensitive as it detected 0.1 pg DNA of T. luwenshuni or T. uilenbergi. Moreover, the assay was evaluated by testing 86 field samples in comparison to the reverse line blot method, showing a calculated sensitivity and specificity of 66.0% and 97.4%, respectively. These results indicate that the LAMP assay has a potential usefulness for application in diagnostic and pidemiological studies on T. luwenshuni and T. uilenbergi infection of small ruminants. In addition, serological screening of the library led to discovery of a positive clone called TuIP, which has been deposited in Genbank under accession number FJ467922. Partially recombinantly cloned and expressed TuIP showed strong reactivity with serum from T. uilenbergi infected animals, indicating its potential usefulness for development of novel serological diagnostic tests or serving as a candidate for vaccine development in the future.

In recent years, Anaplasma phagocytophilum and Rickettsia spp. have been detected in Ixodes ricinus in Germany and a focal distribution has been suggested for A. phagocytophilum. In the present study the prevalence of A. phagocytophilum and spotted fever group (SFG) rickettsiae was investigated in I. ricinus. DNA-extracts were taken from 2,862 unfed I. ricinus ticks (adults and nymphs) from eight sites in Munich, sampled over a five-month period. Single samples from three comparative sites outside of Munich were also included. A real-time PCR targeting the msp2 gene of A. phagocytophilum was used for screening and a nested PCR targeting the 16S rRNA gene for sequencing of 30% of positives. Screening for Rickettsia spp. was performed with a PCR targeting the citrate synthase gene (gltA), followed by PCRs detecting the ompA gene for all gltA positives, and the ompB and 16S rRNA genes for clarifying results of some samples. The overall prevalence was 2.90% (95% CI 2.27 to 3.48%) for A. phagocytophilum and 5.28% (95% CI 4.31 to 6.17%) for SFG rickettsiae. Only 0.31% of the ticks investigated were coinfected. Statistical analysis revealed that prevalence of A. phagocytophilum in ticks from city parks in Munich was significantly higher than in ticks from natural forest, whereas the prevalence of Rickettsia spp. was the opposite. For both, the prevalence in adults was significantly higher than in nymphs. Although wide ranges of prevalence were observed monthly, the variations were not significant along the observational period. Sequence analysis of 16S rRNA PCR products (n=31) revealed 100% homology to Ehrlichia sp. “Frankonia 2”, only one differed in one nucleotide position. All differed in one nucleotide position from the HGA agent detected in human patients. All rickettsial PCR products were also sequenced. All gltA sequences of R. helvetica (n=138) were 100% identical to each other and differed in one nucleotide position from the prototype sequence. Two different R. monacensis strains (n=13) were detected, which differed in up to 4 nucleotide positions in gltA, ompA and ompB. Further rickettsial strains (n=3) most probably belonging to rickettsial endosymbionts were also found. These results show, by molecular methods, a wide distribution of A. phagocytophilum and SFG rickettsiae in I. ricinus ticks in Southern Germany. SFG rickettsiae which are thought to be involved in human disease (R. helvetica and R. monacensis) had a significantly higher prevalence in natural forest areas. Prevalence of A. phagocytophilum was significantly higher in city parks; the genetic strain has not yet been associated with human infection.

In der vorliegenden Arbeit wurden Real-time PCR-basierte Nachweisverfahren für E. canis und A. phagocytophilum entwickelt, validiert und im Anschluss für die Untersuchung von Patientenproben eingesetzt. Für E. canis wurden für zwei Tests Primer und Sonden des Typs „Molecular Beacon“ konstruiert, die auf unterschiedliche Zielgene gerichtet waren, die Reaktionsbedingungen optimiert und die Leistungsfähigkeit beider Tests verglichen. Die PCR EC-16S hatte hierbei die 16S rDNA als Zielgen, während die PCR ECP-p30 auf das p30-10-Gen von E. canis gerichtet war. Bei der Ermittlung der analytischen Sensitivität und analytischen Spezifität ergab sich, dass beide Tests in ihren Leistungen sehr ähnlich waren. Beide PCRs waren spezifisch und lieferten nur für DNA von E. canis ein positives Ergebnis, während die übrigen getesteten Erreger A. platys, N. risticii, A. phagocytophilum, Babesia canis, B. gibsoni und H. canis in der PCR negativ reagierten. Da die PCR ECP-p30 bei der Sensitivitätsprüfung geringfügig besser beurteilt wurde, wurde entschieden, die weiteren Untersuchungen mit diesem PCR-Protokoll durchzuführen. Zur Bestimmung der diagnostischen Sensitivität und Spezifität dieses Tests wurde eine extern kontrollierte Validierung mit geblindeten Proben durchgeführt. Hierbei ergab sich eine diagnostische Spezifität von 100 %. Die diagnostische Sensitivität der Real-time PCR betrug 82 %. Der prädiktive Wert des positiven Testergebnisses lag für die PCR ECP-p30 bei 100 %, während der prädiktive Wert des negativen Testergebnisses 87,5 % erreichte Als Nachweisgrenze wurden 14,5 Moleküle des Zielgens pro 50 µl Ansatz ermittelt. Im Anschluss wurde die Eignung des Tests an Blutproben von Hunden aus einem für E. canis endemischen Gebiet untersucht. Dabei wurden 244 Blutproben einbezogen und die Ergebnisse der PCR mit denen eines IFATs verglichen. Die Blutproben stammten aus Kampanien, Italien und wurden dort durch Tierärzte von Hunden gewonnen, die in einer Tierarztpraxis mit angeschlossenem Tierheim vorgestellt wurden. Die Tiere waren drei Gruppen zuzuordnen: Ein Teil der Hunde, und zwar 19 Tiere, wurde von privaten Besitzern in der Praxis vorgestellt, 52 Tiere waren unmittelbar zuvor von der Strasse aufgelesen worden und die dritte Gruppe, die 173 Hunde umfasste, hielt sich zum Zeitpunkt der Probennahme schon längere Zeit im Tierheim auf. Innerhalb des Tierheims wird ein hoher diagnostischer und medikamenteller Aufwand zur Erkennung und Bekämpfung von E. canis mittels antibiotischer Therapie und Zecken¬prophylaxe betrieben. In die Untersuchung einbezogen wurden jedoch nur Hunde, die mindestens drei Monate lang nicht mehr mit einem gegen E. canis wirksamen Medikament behandelt worden waren. Bei der serologischen Untersuchung der Hunde mittels IFAT ergab sich ein Anteil seropositiver Tiere von insgesamt 41,8 %, der auch bei Betrachtung der drei verschiedenen Gruppen nur wenig variierte. So betrug der Anteil seropositiver Tiere innerhalb der Gruppe der Hunde aus dem Tierheim 43,4 %, während 40,4 % der Straßenhunde und 31,6 % der Tiere in privatem Besitz seropositiv waren. Diese Unterschiede waren nicht signifikant. Ein direkter Erregernachweis mittels Real-time PCR erfolgte bei 13,9 % der untersuchten Tiere. Beim Vergleich der Untersuchungsergebnisse von PCR und IFAT wurde eine Überein¬stimmung bei 61,9 % der untersuchten Proben ermittelt. Bei Betrachtung der einzelnen Hundegruppen lag der Anteil der in der PCR positiven Tiere bei den Straßenhunden mit 23,1 % ungefähr doppelt so hoch wie bei den Tieren in Privatbesitz (10,5 %) oder den Tierheim¬hunden im Tierheim (11,6 %). Der Unterschied zwischen den Straßenhunden und den Tierheimhunden war somit signifikant. Diese Ergebnisse weisen auf ein häufiges Vorkommen von E. canis im Untersuchungsgebiet hin und stützen die Auffassung, dass eine Erreger¬elimination mittels Antibiotikatherapie nur schwer zu erreichen ist. Für A. phagocytophilum wurde in der vorliegenden Studie ebenfalls eine Real-time PCR entwickelt und das Testverfahren unter Einbeziehung zweier bereits publizierter Real-time PCR-Protokolle und zwar von Pusterla et al. (1999a) und von Courtney et al. (2004), validiert. Bei der Entwicklung der Real-time PCR für A. phagocytophilum wurde als Zielgen die 16S rDNA herangezogen, da nur hierfür vergleichbare Sequenzen nahe verwandter Ehrlichienspezies verfügbar waren. Alle drei vorliegenden Testverfahren wurden auf ihre analytische Spezifität und ihre analytische Sensitivität überprüft und zusätzlich im Rahmen einer extern kontrollierten Validierung mittels geblindeter Proben verglichen. Hierbei zeigte sich, dass nur die PCR nach Courtney et al (2004), hier als PCR AP-MSP2 bezeichnet, eine sehr gute Spezifität für A. phagocytophilum besaß. Die anderen Tests lieferten auch für N. risticii und A. platys positive Ergebnisse. Die analytische Sensitivität war bei diesem Test ebenfalls um mindestens eine Zehnerpotenz höher als bei den anderen beiden PCRs. Im Rahmen der Validierung wurde für die PCR AP-MSP2 eine diagnostische Spezifität von 96 % ermittelt, während die im Rahmen dieser Studie entwickelte PCR AP-16S eine Spezifität von 64 % und die PCR nach Pusterla et al. (1999a) einen Wert von 36 % erreichten. Der prädiktive Wert des positiven Testergebnisses betrug für die drei PCRs somit 96 %, 74 % bzw. 61 %. Für die Untersuchung von Patientenproben auf Befall mit A. phagocytophilum wurde deshalb die PCR AP-MSP2 ausgewählt. In die Studie wurden Hunde aus Deutschland einbezogen, und zwar sowohl 72 Blutproben, die eigens für diese Studie auf Anforderung von Tierärzten eingesandt worden waren, als auch 133 Proben, die aus verschiedensten Gründen in das Routinelabor des Institutes eingesandt worden waren. Die 72 eigens für die Studie gewonnenen Proben wurden mittels Buffy-coat-Ausstrich, PCR und IFAT auf A. phagocytophilum untersucht. Lichtmikroskopisch konnten in keinem Fall Einschluss¬körperchen des Erregers in den Granulozyten nachgewiesen werden. Mittels PCR wurde jedoch bei einem Hund (1,4 %) der Nachweis von A. phagocytophilum erbracht. Im IFAT konnten bei 16,7 % der 72 untersuchten Hundeseren spezifische Antikörper gegen den Erreger nachgewiesen werden. Eine Übereinstimmung der Ergebnisse von PCR und Buffy-coat-Ausstrichen lag bei 98,6 % der Proben vor. Beim Vergleich der Ergebnisse der Buffy-coat-Ausstriche mit den Ergebnissen des IFAT wurde eine prozentuale Übereinstimmung von 65,3 % errechnet. Identische Ergebnisse bei PCR und IFAT wurden bei 66,7 % der untersuchten Hunde erzielt. Die 133 Proben, die zufällig aus allen Einsendungen in das Routinelabor des Instituts für vergleichende Tropenmedizin und Parasitologie ausgewählt worden waren, wurden mittels PCR und IFAT untersucht, wobei zwei Tiere (1,5 %) in der PCR und 34,6 % im IFAT ein positives Ergebnis lieferten. Die Identität des PCR-Produktes eines der positiven Tiere wurde durch Klonierung und anschließende Sequenzierung bestätigt. Eine Übereinstimmung der Testergebnisse von IFAT und PCR bestand bei 52,6 % der untersuchten Proben. Diese Ergebnisse stützen die Auffassung, dass Hunde in Deutschland häufig mit A. phagocytophilum in Kontakt kommen, dass es sich dabei aber meist um eine selbstlimitierende, klinisch inapparente Infektion handelt.

625 adult unfed I. ricinus ticks from three recreational areas located near Munich and Passau, 275 adult engorged I. ricinus ticks from dogs of Bavaria and Baden-Württemberg and 25 adult engorged I. ricinus ticks from cattle of an area in Switzerland, which is endemic for bovine granulocytic ehrlichiosis, were collected over a specific period (3/2003-3/2004). The ticks were examined for an infection with B. burgdorferi sensu lato spp., A. phagocytophilum and piroplasms by the use of Real-Time PCR and nested PCR. In addition RFLP analysis and sequencing were chosen for the differentiation of the species and OspA types of B. burgdorferi sensu lato. The examination of unfed ticks resulted in a prevalence of 35,4% for B. burgdorferi sensu lato, 4,5% for A. phagocytophilum and 1,3% for piroplasms. There was no significant difference for the infection rates of B. burgdorferi sensu lato between the different sampling areas, whereas A. phagocytophilum showed a significantly higher prevalence in one sampling side in Munich and a significantly lower prevalence in Passau. Apart from infections with only one pathogen, coinfections with B. burgdorferi sensu lato and A. phagocytophilum could be detected in 1,1% of the unfed ticks, with a local cluster in one area in Munich and 0,3% of the unfed ticks showed a coinfection with B. burgdorferi sensu lato and piroplasms. Prevalence rates of 8,4%, 4,7% and 5,1% were identified for B. burgdorferi sensu lato, A. phagocytophilum and piroplasms respectively in engorged ticks from dogs, 0,7% of these ticks were coinfected with B. burgdorferi sensu lato and A. phagocytophilum. The examination of engorged ticks from cattle revealed in a prevalence rate of 8,0% for B. burgdorferi sensu lato and 60,0% for A. phagocytophilum. The high infection rate of A. phagocytophilum probably resulted from an infection of the cattle with this pathogen. The difference in the prevalence rate of B. burgdorferi sensu lato in unfed and engorged ticks might be caused by their distinct geographical origin, the degree of blood uptake and different factors in the blood of the different host species which are able to protect the host from being infected with Borrelia. The differentiation of B. burgdorferi sensu lato into the species and OspA types showed that the clinically relevant species B. afzelii, B. burgdorferi sensu stricto and B. garinii, in which B. garinii was represented by the OspA types 3, 4, 5, 6 and 8 could be detected. Additionally, B. valaisiana, a species which is suspected of being pathogenic to humans and the recently described new Borrelia genospecies, B. spielmanii (previously A14S), were detected. Alltogether a broad heterogeneity for Borrelia species and subspecies (classified by OspA types) could be observed in unfed ticks, above all in one sampling side in Munich. Ticks engorgd from dogs and cattle showed a less heterogeneous pattern of distribution.