Oral Azithromycin Versus Doxycycline in the Treatment of Children With Uncomplicated Scrub Typhus: A Randomized Controlled Trial.

OBJECTIVES: To compare the efficacy of azithromycin versus doxycycline in treatment of children with uncomplicated scrub typhus in terms of percentage of children who attained remission of fever after 72 hours of administration of first dose of the study drug, mean time taken to attain fever defervescence, normalization of laboratory parameters, resolution of hepatosplenomegaly and lymphadenopathy. DESIGN: Interventional, open-labeled randomized controlled trial. STUDY METHODS: Patients admitted with undifferentiated fever in the In-Patient Department (IPD), Department of Paediatrics, All India Institute of Medical Sciences, Bhubaneswar, India, as per the inclusion criteria were randomized and was treated with azithromycin at 10 mg/kg/d in one group and doxycycline at 4.4 mg/kg/d for 5 days in the other group and was assessed based on the primary and secondary objectives. RESULTS: There was no statistically significant difference between the percentage of children who attained remission of fever after 72 hours of administration of azithromycin (98.2%) and doxycycline (96.5%) (P value 0.47) and the average time taken for fever defervescence (azithromycin: 24.53 hours; doxycycline: 25.82 hours; P value 0.36). The odds of attaining fever remission in the doxycycline group as compared with the azithromycin group was 1.01 (95% confidence interval -0.60 to -1.71), which was also statistically not significant. There was less incidence of adverse drug events in the azithromycin group (1.78%) as compared with the doxycycline group (8.6%), which was statistically significant (P value 0.02). CONCLUSION: Azithromycin is equally efficacious in terms of fever defervescence, resolution of clinical signs and laboratory parameters as doxycycline, is safer and better tolerated in children.

Mapping the distribution and risk factors of Anaplasmataceae in wild and domestic canines in Chile and their association with Rhipicephalus sanguineus species complex lineages.

Anaplasma platys and Ehrlichia canis are members of the Anaplasmataceae family that cause disease in dogs and are mainly transmitted by Rhipicephalus sanguineus species group ticks. We performed a cross-sectional study on these pathogens across six bioclimatic regions of Chile, including 719 free-ranging rural dogs, 132 Andean foxes (Lycalopex culpaeus), and 82 South American gray foxes (Lycalopex griseus). Dog and fox blood samples were first screened for DNA of Anaplasmataceae followed by two Ehrlichia-specific protocols. Antibodies against Anaplasma sp. and E. canis were assessed by immunofluorescence in dogs. Ectoparasites were collected and identified, with the determination of the lineages of the Rhipicephalus sanguineus species group by molecular and phylogenetic analyses. Finally, potential risk factors for infection were investigated across the different bioclimatic regions and host species. All DNA amplicons obtained from the screening protocol corresponded to Anaplasma platys. The occurrence of both A. platys DNA and antibodies was confirmed in all six bioclimatic regions, except for regions at high altitude and/or without either R. sanguineus species group lineage present. Dogs infested with R. sanguineus ticks were significantly more prone to be infected and exposed to Anaplasma spp. Prevalence of DNA was significantly higher in juvenile (19%) than in adult dogs (9%), whereas the opposite was found for seroprevalence (19% versus 35%, respectively). Overall prevalence of A. platys DNA was higher in dogs (11%) than in foxes (4%), probably owing to markedly lower tick infestations in the foxes. Ehrlichia canis DNA was not detected in any sample, and antibodies against this pathogen were detected only in four dogs, in areas with both R. sanguineus lineages present. Free-ranging dogs in Chile could be favoring the maintenance of A. platys in all areas suitable for its tick vector. Although apparently infrequent, spillovers from dogs to foxes may be taking place and should be considered in management plans in Chile.

Leptospirosis y rickettsiosis, reto diagnóstico para el síndrome febril en zonas endémicas / Leptospirosis and rickettsiosis, a diagnostic challenge for febrile syndrome in endemic areas

Resumen. Se presenta el caso de un hombre de 50 años de edad proveniente de la región de Urabá, Colombia, con una infección mixta por Rickettsia rickettsii y Leptospira interrogans serovar Copenhageni ST78, y pruebas negativas para malaria y dengue. El paciente presentó un síndrome febril que no mejoró con el tratamiento antibiótico sistémico y, finalmente, falleció en la unidad de cuidados intensivos. El diagnóstico post mortem se hizo mediante tipificación molecular de los dos agentes etiológicos. En la inspección del domicilio del paciente, se encontró un ejemplar de Rattus rattus infectado con L. interrogans del mismo serovar detectado en él. No se encontraron garrapatas en los animales domésticos que habitaban con el paciente. Se reporta una infección mixta con síntomas clínicos progresivos y fatales en un paciente con antecedentes laborales de riesgo en una zona endémica para enfermedades tropicales, lo que obliga a tener presente la posibilidad de infecciones simultáneas en personas procedentes de áreas endémicas que consulten reiteradamente por síndrome febril sin resolución y tengan riesgo laboral relacionado con actividades agrícolas.

Abstract. This is the case of a 50-year-old male from the region of Urabá, Colombia, with a mixed infection by Rickettsia rickettsii and Leptospira interrogans serovar Copenhageni ST78 and negative test for malaria and dengue fever. The patient presented with febrile syndrome and was unresponsive to systemic antibiotic treatment, who finally died in the intensive care unit. We established the postmortem diagnosis through molecular typification of the two etiological agents. In the inspection at the patient's home, we found a Rattus rattus specimen infected with L. interrogans of the same serovar found in him. We found no ticks parasitizing the domestic animals cohabitating with the patient. This case of a mixed infection with progressive and fatal symptoms in a patient with occupational risk in a tropical disease endemic zone highlights the importance of considering the potential presentation of simultaneous etiologies in patients with multiple medical visits for unresolved febrile syndromes associated with risky exposure during agricultural activities.

Human seroepidemiology of Rickettsia and Orientia species in Chile - A cross-sectional study in five regions.

In recent years, the spectrum and epidemiology of human rickettsioses has become an emerging topic in Chile. This survey aimed to assess the seroprevalence of spotted fever group rickettsiae (SFGR), typhus group rickettsiae (TGR), and scrub typhus group orientiae (STGO) in northern, central, and southern Chile. We performed a cross-sectional study of healthy adults in rural and urban settings of five regions. Participants were chosen by double stratified random sampling in urban and by convenience in rural locations (n = 1302). Serum specimens were analyzed for group-specific IgG antibodies against SFGR, TGR, and STGO by enzyme-linked immunosorbent assays (ELISAs). Overall seroprevalences to SFGR, TGR, and STGO were 5.3 %, 1.2 %, and 0.4 %, respectively. Prevalences showed geographical differences. Statistical analyses revealed an association of older age with seropositivity to SFGR and to TGR and of rural setting and male gender with seropositivity to SFGR. The study indicates that SFGR, TGR, and STGO are endemic in Chile. The very low STGO seroprevalence might indicate an insufficient sensitivity of serological tests using Asian O. tsutsugamushi strains as ELISA antigens for the detection of antibodies against Chilean Orientia species.

Aspectos clínico-epidemiológicos de um cluster de febre maculosa brasileira, ocorrido no município de Americana, São Paulo, Brasil, 2018 / Clinical and epidemiological aspects of a cluster of brazilian spotted fever, which occurred in the city of Americana, São Paulo, Brazil, 2018

Objetivos: descrever os principais aspectos clínicos, laboratoriais e epidemiológicos relacionados à ocorrência de um cluster de febre maculosa brasileira (FMB) na cidade de Americana/SP, em 2018. Métodos: realizou-se uma análise descritiva baseada na revisão das informações constantes nos registros do Sistema de Informação de Agravos de Notificação ­ SINAN e nos relatórios de investigações epidemiológicas obtidos junto ao banco de dados da Unidade de Vigilância em Saúde Municipal. Resultados: esse cluster ocorreu em um fragmento de mata ciliar situado na confluência dos rios Atibaia e Jaguari, entre os dias 21/4/2018 e 31/5/2018. Durante esse período, ocorreram 15 casos positivos de FMB, com registro de 11 óbitos, atingindo uma taxa de letalidade de 73%. O sexo masculino representou 93% dos pacientes, e a faixa etária mais afetada foi a de indivíduos acima dos 30 anos (67%), seguindo-se o percentual de 13% para as faixas de 1 a 4 e 5 a 9 anos, e 7% para pessoas de 20 a 29 anos. Os principais sinais clínicos observados foram febre (100%); cefaléia (80%) e mialgia (73,33%). Conclusão: foi possível delinear o perfil epidemiológico dos indivíduos a serem alcançados nas campanhas de educação em saúde, com enfoque para medidas preventivas visando à redução de frequência nas áreas de risco para parasitismo humano por carrapatos vetores da FMB.

Objective: describe the main clinical, laboratory and epidemiological aspects related to the occurrence of a cluster of brazilian spotted fever (FMB) in the city of Americana/SP, in 2018. Methods: A descriptive analysis was performed based on the review of the information contained in the records of the Notification Disease Information System (SINAN) and in the reports of epidemiological investigation obtained from the Municipal Health Surveillance Unit database. Results: This cluster occurred in a riparian forest fragment located at the confluence of the Atibaia and Jaguari rivers, between 04/21/2018 and 05/31/2018. During this period there were 15 positive cases of BSF, with a record of 11 deaths, reaching a lethality rate of 73%. Males represented 93% of the patients and the most affected age group was that of individuals over 30 years old (67%), followed by 13% for the age groups 01 to 04 and 05 to 09 years old, and 7 % for people from 20 to 29 years old. The main clinical signs observed were fever (100,00%); headache (80,00%) and myalgia (73,33%). Conclusion: It was possible to outline an epidemiological profile of individuals to be reached in health education campaigns, with a focus on preventive measures aimed at reducing frequency in areas at risk for human parasitism by BSF vector ticks.

A review on the occurrence of companion vector-borne diseases in pet animals in Latin America.

Companion vector-borne diseases (CVBDs) are an important threat for pet life, but may also have an impact on human health, due to their often zoonotic character. The importance and awareness of CVBDs continuously increased during the last years. However, information on their occurrence is often limited in several parts of the world, which are often especially affected. Latin America (LATAM), a region with large biodiversity, is one of these regions, where information on CVBDs for pet owners, veterinarians, medical doctors and health workers is often obsolete, limited or non-existent. In the present review, a comprehensive literature search for CVBDs in companion animals (dogs and cats) was performed for several countries in Central America (Belize, Caribbean Islands, Costa Rica, Cuba, Dominican Republic, El Salvador, Guatemala, Honduras, Mexico, Nicaragua, Panama, Puerto Rico) as well as in South America (Argentina, Bolivia, Brazil, Chile, Colombia, Ecuador, French Guiana, Guyana (British Guyana), Paraguay, Peru, Suriname, Uruguay, Venezuela) regarding the occurrence of the following parasitic and bacterial diseases: babesiosis, heartworm disease, subcutaneous dirofilariosis, hepatozoonosis, leishmaniosis, trypanosomosis, anaplasmosis, bartonellosis, borreliosis, ehrlichiosis, mycoplasmosis and rickettsiosis. An overview on the specific diseases, followed by a short summary on their occurrence per country is given. Additionally, a tabular listing on positive or non-reported occurrence is presented. None of the countries is completely free from CVBDs. The data presented in the review confirm a wide distribution of the CVBDs in focus in LATAM. This wide occurrence and the fact that most of the CVBDs can have a quite severe clinical outcome and their diagnostic as well as therapeutic options in the region are often difficult to access and to afford, demands a strong call for the prevention of pathogen transmission by the use of ectoparasiticidal and anti-feeding products as well as by performing behavioural changes.

Respecto a un caso de rickettsia rickettsii en Costa Rica / Regarding a rickettsia rickettsii case in Costa Rica

Resumen Las rickettsiosis representan una zoonosis de etiología bacteriana intracelular, las cuales son transmitidas por vectores como artrópodos a la piel y luego a las células endoteliales microvasculares. La Fiebre de las Montañas Rocosas (FMMR) causada por Rickettsiarickettsiies la más grave, y es una enfermedad reportada en Costa Rica desde 1977, que causa daño endotelial directo asociado a liberación de citocinasproinflamatorias y estrés oxidativo, lo que lleva a un proceso de vasculitis de pequeño y mediano vaso. La enfermedad se manifiesta clínicamente por fiebre acompañada de cefalea y mialgias, síntomas gastrointestinales y un brote cutáneo. El diagnóstico es basado en la clínica, la exposición a vectores y el respaldo de ensayos de laboratorio; y las tetraciclinas son su tratamiento de elección. El objetivo del presente trabajo es documentar un caso clínico confirmado de Fiebre Manchada de las Montañas Rocosas en Costa Rica, para evidenciar la importancia de considerar esta entidad como un diagnóstico diferencial en un síndrome febril agudo.

Abstract Rickettsiosis represents a zoonosis of intracellular bacterial etiology, which is transmitted by vectors such as arthropods to the skin and then to microvascular endothelial cells. Rocky Mountain Fever caused by Rickettsia rickettsii is the most serious, and it is a disease reported in Costa Rica since 1977. It causes direct endothelial damage associated with the release of proinflammatory cytokines and oxidative stress, which leads to a vasculitis of the small and medium vessels. The disease is manifested clinically by fever headache, myalgias, gastrointestinal symptoms and a cutaneous rash. The diagnosis is based on clinical manifestations, exposure to vectors and the support of laboratory tests; and tetracyclines are the treatment of choice. The objective of this work is to document a case of a Rocky Mountain Spotted Fever in Costa Rica, to demonstrate the importance of considerate this entity as a differential diagnosis in an acute febrile syndrome.

Serological Evidence of Rickettsia, Orientia, and Coxiella in Domestic Animals from Bhutan: Preliminary Findings.

There is no information on rickettsial diseases in domestic animals in Bhutan. This study provides preliminary serological data on exposure of domestic animals to Rickettsia, Orientia, and Coxiella. Animal sera were collected opportunistically from Bhutan and tested in the Australian Rickettsial Reference Laboratory for IgG antibodies against spotted fever group (SFG) and typhus group (TG) Rickettsia, scrub typhus group (STG), and Q fever (QF). Of the 294 animals tested, 136 (46%) showed serological evidence of past exposure to one or more rickettsiae: 106 (36%), 62 (21%), 45 (15%), and 11 (4%) being positive against SFG Rickettsia, Orientia, TG Rickettsia, and Coxiella, respectively. Dogs appeared to exhibit the highest seropositivity against SFG (55%) and TG Rickettsia (45%), horses against STG (91%), while goats were mostly positive for Coxiella (9%). Dogs also appeared to have high risk of being exposed to SFG Rickettsia (odd ratios [OR] 5.71, 95% confidence interval [CI] 3.02-10.80, p < 0.001), TG Rickettsia (OR 48.74, 95% CI 11.29-210.32, p < 0.001), and STG (OR 6.80, 95% CI 3.32-13.95, p < 0.001), but not against QF (OR 1.95, 95% CI 0.42-8.95, p = 0.390). Differences in seropositivity rates between animal species may have been significant for SFG, TG, and STG, but not for QF. The differences in the seropositivity rates of the four infections between districts appeared to be significant for TG and STG, but not for SFG and QF. The seropositivity rates of domestic animals to the four rickettsial infections were consistent with similar studies on the human population in the same areas and appear to demonstrate a high prevalence of exposure to rickettsiae in Bhutan. These preliminary findings constitute baseline data for Bhutan. The findings of this study call for an increased human-livestock sector collaboration in rickettsial diseases research aimed at developing diagnostic and therapeutic guidelines and formulating preventive and control measures through a One Health approach.

Sociocultural descriptions of febrile syndromes in rural areas of Urabá, Colombia: An exploration of "tick fever" / Descrição sociocultural de síndromes febris em áreas rurais de Antioquia Urabá: um olhar sobre a "febre do carrapato"

Abstract Introduction: In 2006 and 2008 there were two lethal outbreaks of rickettsioses in the rural areas of Urabá, characterized by the lack of immediate diagnosis and antibiotic treatment. Objective: Describe sociocultural aspects about knowledge, attitudes and practices in relation to febrile syndromes and "tick fever" in rural areas of Urabá. Materials and methods: We conducted an exploratory study using knowledge, attitudes, and practices questionnaires and semi-structured interviews about febrile syndromes and "tick fever". We surveyed 246 heads of households and interviewed nine individuals. Results: We observed that people tended to identify febrile syndromes with signs and clinical symptoms of dengue, malaria, leptospirosis and rickettsioses. A considerable proportion of individuals (32.93%) knew very little about "tick fever", thinking that is was transmitted by mosquitos. They mentioned intestinal parasitoids, malaria, dengue, and "evil eye" among the causes of febrile syndromes. "Tick fever" is linked by its name to the bite of the tick. Furthermore, the treatments for febrile syndromes mentioned by interviewees are associated to those commonly used in western medicine and medicinal plants. Conclusions: There is a need for educational programs in rural areas, to raise awareness about these potential lethal conditions that can be effectively treated.

Resumo Introdução: Nos anos de 2006 e 2008, dois surtos letais de riquetsiose foram relatados em áreas rurais de Urabá, caracterizados pela falta de atenção imediata para o diagnóstico e tratamento com antibióticos. Objetivo: Descrever aspectos socioculturais do conhecimento, atitudes e práticas de síndromes febris e "febre do carrapato" em áreas rurais de Antioquia, Urabá. Materiais e métodos: Realizou-se estudo exploratório, através de inquéritos sobre CAP (conhecimentos, atitudes e práticas) e entrevistas semiestruturadas sobre síndromes febris e "febre do carrapato", um nome que se refere às doenças das riquetsioses na área. Foram aplicados 246 inquéritos sobre a CAP foram realizadas aos chefes de agregados familiares e nove pessoas foram entrevistadas. Resultados: Observou-se que as pessoas tendem a identificar os sinais e sintomas clínicos característicos da dengue, malária, leptospirose e até riquetsiose. Um grande número de pessoas demonstra baixo conhecimento sobre "febre do carrapato" e indicou que ela é transmitida por mosquitos (32,93%). Os entrevistados indicaram que as causas das diferentes síndromes febris se devem a parasitas intestinais, malária, dengue ou, como crença cultural, ao "mau olhado". A "febre do carrapato" o associa, pelo nome, com a mordida do carrapato. Além disso, o tratamento das síndromes febris referidas pelos entrevistados está intimamente relacionado ao uso comum da medicina ocidental e ao uso de "plantas medicinais". Conclusões. É necessário ter programas de educação nessas áreas rurais para que essas entidades potencialmente letais tenham um tratamento efetivo e acessível.

Garrapatas y enfermedades transmitidas por garrapatas en la reserva ecológica costanera sur, ciudad autónoma de Buenos Aires / Ticks and diseases transmitted by ticks in the southern coastal ecological reserve, autonomous city of Buenos Aires

INTRODUCCIÓN Las enfermedades zoonóticas transmitidas por garrapatas (tales como rickettsiosis y borreliosis) son consideradas enfermedades emergentes y reemergentes en numerosos lugares del mundo, incluso en Argentina. La Reserva Ecológica Costanera Sur es un área urbana protegida ubicada en la Ciudad Autónoma de Buenos Aires que se caracteriza por una gran cantidad de ambientes de origen artificial, como bañados, lagunas, pastizales, matorrales y bosques, además de las playas del Río de la Plata. OBJETIVOS Estudiar los aspectos eco-epidemiológicos de las garrapatas y de los microorganismos patógenos transmitidos por garrapatas en la Reserva Ecológica Costanera Sur de la Ciudad Autónoma de Buenos Aires en el período 2011-2015. MÉTODOS Se realizaron muestreos para recolectar garrapatas de vegetación y de hospedadores como perros y distintas especies de roedores. Mediante técnicas moleculares se buscó la presencia de microorganismos de los géneros Rickettsia (fragmento del ARNr 23S-5S) y Borrelia (fragmento del ARNr 16S). RESULTADOS En total se recolectaron 1090 garrapatas de la vegetación (454 de la especie Amblyomma aureolatum, 653 Ixodes auritulus y 1 Amblyomma triste) y 67 de los perros (64 A. aureolatum, 2 Rhipicephalus sanguineus sensu lato y 1 A. triste). No se detectaron garrapatas en los 201 roedores revisados. Mediante PCR se estudiaron 899 garrapatas (375 A. aureolatum, 523 I. auritulus y 1 A. triste), y se detectó un 8,3% de I. auritulus positivos al género Borrelia. Todas las garrapatas resultaron negativas al género Rickettsia. El producto amplificado positivo a Borrelia pudo ser secuenciado, con un 97,9% de identidad respecto a distintas secuencias del complejo Borrelia burgdorferi sensu lato halladas en garrapatas del género Ixodes en el hemisferio norte. DISCUSIÓN La presencia abundante de las garrapatas A. aureolatum e I. auritulus, así como la detección del complejo B. burgdorferi s. l. en la región resultan de suma importancia para la salud pública.

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping.

The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein (wsp) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (ftsZ-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima, which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima. Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

Vector species-specific association between natural Wolbachia infections and avian malaria in black fly populations.

Artificial infection of mosquitoes with the endosymbiont bacteria Wolbachia can interfere with malaria parasite development. Therefore, the release of Wolbachia-infected mosquitoes has been proposed as a malaria control strategy. However, Wolbachia effects on vector competence are only partly understood, as indicated by inconsistent effects on malaria infection reported under laboratory conditions. Studies of naturally-occurring Wolbachia infections in wild vector populations could be useful to identify the ecological and evolutionary conditions under which these endosymbionts can block malaria transmission. Here we demonstrate the occurrence of natural Wolbachia infections in three species of black fly (genus Simulium), which is a main vector of the avian malaria parasite Leucocytozoon. Prevalence of Leucocytozoon was high (25%), but the nature and magnitude of its association with Wolbachia differed between black fly species. Wolbachia infection was positively associated with avian malaria infection in S. cryophilum, negatively associated in S. aureum, and unrelated in S. vernum. These differences suggest that Wolbachia interacts with the parasite in a vector host species-specific manner. This provides a useful model system for further study of how Wolbachia influences vector competence. Such knowledge, including the possibility of undesirable positive association, is required to guide endosymbiont based control methods.

Conflict in the Intracellular Lives of Endosymbionts and Viruses: A Mechanistic Look at Wolbachia-Mediated Pathogen-blocking.

At the forefront of vector control efforts are strategies that leverage host-microbe associations to reduce vectorial capacity. The most promising of these efforts employs Wolbachia, a maternally transmitted endosymbiotic bacterium naturally found in 40% of insects. Wolbachia can spread through a population of insects while simultaneously inhibiting the replication of viruses within its host. Despite successes in using Wolbachia-transfected mosquitoes to limit dengue, Zika, and chikungunya transmission, the mechanisms behind pathogen-blocking have not been fully characterized. Firstly, we discuss how Wolbachia and viruses both require specific host-derived structures, compounds, and processes to initiate and maintain infection. There is significant overlap in these requirements, and infection with either microbe often manifests as cellular stress, which may be a key component of Wolbachia's anti-viral effect. Secondly, we discuss the current understanding of pathogen-blocking through this lens of cellular stress and develop a comprehensive view of how the lives of Wolbachia and viruses are fundamentally in conflict with each other. A thorough understanding of the genetic and cellular determinants of pathogen-blocking will significantly enhance the ability of vector control programs to deploy and maintain effective Wolbachia-mediated control measures.

Tick- and fly-borne bacteria in ungulates: the prevalence of Anaplasma phagocytophilum, haemoplasmas and rickettsiae in water buffalo and deer species in Central Europe, Hungary.

BACKGROUND: Hunting constitutes an important industry in Europe. However, data on the prevalence of vector-borne bacteria in large game animal species are lacking from several countries. Blood or spleen samples (239 and 270, respectively) were taken from red, fallow and roe deer, as well as from water buffaloes, mouflons and wild boars in Hungary, followed by DNA extraction and molecular analyses for Anaplasma phagocytophilum, haemoplasmas and rickettsiae. RESULTS: Based on blood samples, the prevalence rate of A. phagocytophilum infection was significantly higher in red deer (97.9%) than in fallow deer (72.7%) and roe deer (60%), and in all these compared to mouflons (6.3%). In addition, 39.2% of the spleen samples from wild boars were PCR positive for A. phagocytophilum, but none of the buffalos. Based on blood samples, the prevalence rates of both Mycoplasma wenyonii (Mw) and 'Candidatus M. haemobos' (CMh) infections were significantly higher in buffaloes (Mw: 91.2%; CMh: 73.3%) than in red deer (Mw: 64.6%; CMh: 45.8%), and in both of them compared to fallow deer (Mw: 30.3%; CMh: 9.1%) and roe deer (Mw: 20%; CMh: 1.5%). The prevalence of Mw and CMh infection significantly correlated with the body sizes of these hosts. Furthermore, Mw was significantly more prevalent than CMh in buffaloes, red and roe deer. Mycoplasma ovis was detected in mouflons, M. suis in wild boars, R. helvetica in one fallow deer and one mouflon, and an unidentified Rickettsia sp. in a fallow deer. CONCLUSIONS: Forest-dwelling game animal species were found to be important carriers of A. phagocytophilum. In contrast, animals grazing grassland (i.e. buffaloes) were less likely to get infected with this Ixodes ricinus-borne pathogen. Water buffaloes, deer species, mouflons and wild boars harbored haemoplasmas that may affect domestic ungulates. Evaluated animals with larger body size had significantly higher prevalence of infection with haemoplasmas compared to smaller deer species. The above host species rarely carried rickettsiae.

Utilidad del diagnóstico molecular precoz de fiebre Q y rickettsiosis en pacientes con fiebre de duración intermedia / Usefulness of the early molecular diagnosis of Q fever and rickettsial diseases in patients with fever of intermediate duration

La mayor parte de los casos de fiebre de duración intermedia (FDI) en España corresponden a enfermedades infecciosas (principalmente fiebre Q y rickettsiosis). En la práctica clínica el diagnóstico causal de estas entidades se basa en el inmunodiagnóstico, con una escasa utilidad en fases precoces. Por ello, el objetivo de este trabajo fue la evaluación de la utilidad de técnicas moleculares en el diagnóstico precoz de fiebre Q y rickettsiosis en pacientes con FDI. Se estudió mediante PCR la presencia de material genético de Coxiella burnetii y Rickettsia spp. en muestras sanguíneas de 271 pacientes con FDI. La especificidad de ambas técnicas es elevada, permitiendo el diagnóstico en casos no diagnosticados mediante detección de anticuerpos específicos. Estos datos sugieren que el empleo de técnicas moleculares, con una adecuada selección de la muestra de estudio y el empleo de cebadores adecuados, es un elemento útil en el diagnóstico precoz de las principales causas de FDI, principalmente si la serología es negativa o no es concluyente (AU)

Most cases of fever of intermediate duration (FDI) in Spain are associated with infectious diseases (mainly Q fever and rickettsia infections). In clinical practice, the causal diagnosis of these entities is based on immunodiagnostic techniques, which are of little help in the early stages. Therefore, the aim of this study was to evaluate the usefulness of molecular techniques for the early diagnosis of Q fever and rickettsia diseases in patients with FDI. A PCR method was used to detect the presence of genetic material of Coxiella burnetii and Rickettsia spp. in blood specimens from 271 patients with FDI. The specificity of both techniques is high, allowing diagnosis in cases undiagnosed by specific antibodies detection. These data suggest that the use of molecular techniques, with proper selection of the study specimen, and using appropriate primers is a useful tool in the early diagnosis of the main causes of FDI, especially if serology is negative or inconclusive (AU)

Molecular detection of Anaplasma platys, Anaplasma phagocytophilum and Wolbachia sp. but not Ehrlichia canis in Croatian dogs.

The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.