Integrated analysis of the sialotranscriptome and sialoproteome of the rat flea Xenopsylla cheopis.

Over the last 20 years, advances in sequencing technologies paired with biochemical and structural studies have shed light on the unique pharmacological arsenal produced by the salivary glands of hematophagous arthropods that can target host hemostasis and immune response, favoring blood acquisition and, in several cases, enhancing pathogen transmission. Here we provide a deeper insight into Xenopsylla cheopis salivary gland contents pairing transcriptomic and proteomic approaches. Sequencing of 99 pairs of salivary glands from adult female X. cheopis yielded a total of 7432 coding sequences functionally classified into 25 classes, of which the secreted protein class was the largest. The translated transcripts also served as a reference database for the proteomic study, which identified peptides from 610 different proteins. Both approaches revealed that the acid phosphatase family is the most abundant salivary protein group from X. cheopis. Additionally, we report here novel sequences similar to the FS-H family, apyrases, odorant and hormone-binding proteins, antigen 5-like proteins, adenosine deaminases, peptidase inhibitors from different subfamilies, proteins rich in Glu, Gly, and Pro residues, and several potential secreted proteins with unknown function. SIGNIFICANCE: The rat flea X. cheopis is the main vector of Yersinia pestis, the etiological agent of the bubonic plague responsible for three major pandemics that marked human history and remains a burden to human health. In addition to Y. pestis fleas can also transmit other medically relevant pathogens including Rickettsia spp. and Bartonella spp. The studies of salivary proteins from other hematophagous vectors highlighted the importance of such molecules for blood acquisition and pathogen transmission. However, despite the historical and clinical importance of X. cheopis little is known regarding their salivary gland contents and potential activities. Here we provide a comprehensive analysis of X. cheopis salivary composition using next generation sequencing methods paired with LC-MS/MS analysis, revealing its unique composition compared to the sialomes of other blood-feeding arthropods, and highlighting the different pathways taken during the evolution of salivary gland concoctions. In the absence of the X. cheopis genome sequence, this work serves as an extended reference for the identification of potential pharmacological proteins and peptides present in flea saliva.

Molecular detection of Rickettsia in ectoparasites (Siphonaptera and Phthiraptera) of domestic and feral pigs from Argentina.

Rickettsioses are distributed among a variety of hematophagous arthropods, and represent an emergent threat. The presence of rickettsial bacteria in ectoparasites collected from pigs from Argentina is still unknown. This study investigated the presence and identity of Rickettsia spp. in fleas, Pulex irritans, and sucking lice, Haematopinus suis, of domestic and feral pigs, Sus scrofa, from Central-Northern Argentina, through the genes gltA and ompB. Rickettsial bacteria were detected in 50% of fleas and 24% of lice. The BLASTn analysis of the ompB gene fragments in P. irritans samples showed identities 99% and 100% with R. felis. Positive samples of H. suis were 99% similar with species from the spotted fever group, future amplifications of a more polymorphic fragment of the ompB gene will allow to corroborate the identity of the Rickettsia species present in these lice samples. The Rickettsia spp. reported in the present study are having eventually been associated with cases of human diseases, and the circulation of these agents in arthropods has already been reported in several countries. Therefore, the identification of circulating pathogenic agents, such as reported in this study, is crucial for development of preventive measures for the control of ectoparasite-borne rickettsiosis diseases. Further studies, using serology techniques, will be allow to explore the ability of pigs as a possible Rickettsia reservoir and its role as part of transmission cycle of Rickettsia spp. in the studied scenarios.

The genus Rickettsia in Mexico: Current knowledge and perspectives.

The genus Rickettsia encompasses 35 valid species of intracellular, coccobacilli bacteria that can infect several eukaryotic taxa, causing multiple emerging and re-emerging diseases worldwide. This work aimed to gather and summarise the current knowledge about the genus Rickettsia in Mexico, updating the taxonomy of the bacteria and their hosts by including all the records available until 2020, to elucidate host-parasite relationships and determine the geographical distribution of each Rickettsia species present in the country. Until now, 14 species of Rickettsia belonging to four groups have been recorded in Mexico. These species have been associated with 26 arthropod species (14 hard ticks, three soft ticks, two sucking lice, and seven fleas) and 17 mammal species distributed over 30 states in Mexico. This work highlights the high biological inventory of rickettsias for Mexico and reinforces the need to approach the study of this group from a One Health perspective.

Yersinia pestis: the Natural History of Plague.

The Gram-negative bacterium Yersinia pestis is responsible for deadly plague, a zoonotic disease established in stable foci in the Americas, Africa, and Eurasia. Its persistence in the environment relies on the subtle balance between Y. pestis-contaminated soils, burrowing and nonburrowing mammals exhibiting variable degrees of plague susceptibility, and their associated fleas. Transmission from one host to another relies mainly on infected flea bites, inducing typical painful, enlarged lymph nodes referred to as buboes, followed by septicemic dissemination of the pathogen. In contrast, droplet inhalation after close contact with infected mammals induces primary pneumonic plague. Finally, the rarely reported consumption of contaminated raw meat causes pharyngeal and gastrointestinal plague. Point-of-care diagnosis, early antibiotic treatment, and confinement measures contribute to outbreak control despite residual mortality. Mandatory primary prevention relies on the active surveillance of established plague foci and ectoparasite control. Plague is acknowledged to have infected human populations for at least 5,000 years in Eurasia. Y. pestis genomes recovered from affected archaeological sites have suggested clonal evolution from a common ancestor shared with the closely related enteric pathogen Yersinia pseudotuberculosis and have indicated that ymt gene acquisition during the Bronze Age conferred Y. pestis with ectoparasite transmissibility while maintaining its enteric transmissibility. Three historic pandemics, starting in 541 AD and continuing until today, have been described. At present, the third pandemic has become largely quiescent, with hundreds of human cases being reported mainly in a few impoverished African countries, where zoonotic plague is mostly transmitted to people by rodent-associated flea bites.

Immunity of fleas (Order Siphonaptera).

The immune response of arthropod vectors plays a key role in the spread and transmission of vector-borne diseases. Although fleas transmit several human pathogens (e.g., Bartonella henselae, Rickettsia felis, R. typhi, and Yersinia pestis), few studies have examined how these vectors respond to infection. In hematophagous arthropods, imbibed pathogens must survive the hostile environment of blood meal digestion, which includes proteolytic digestive enzymes, protease inhibitors and expression of genes associated with protection of epithelial linings. Additionally, insect epithelial cells exhibit local immune defense against ingested pathogens by producing antimicrobial peptides and reactive oxygen species. This review details these and other aspects of insect immunity as it relates to fleas, with an emphasis on the gut immune response to two blood-borne pathogens, R. typhi and Y. pestis.

A Single Amino Acid Change in the Response Regulator PhoP, Acquired during Yersinia pestis Evolution, Affects PhoP Target Gene Transcription and Polymyxin B Susceptibility.

Yersinia pestis, the causative agent of plague, evolved from the closely related pathogen Yersinia pseudotuberculosis During its emergence, Y. pestis is believed to have acquired its unique pathogenic characteristics through numerous gene gains/losses, genomic rearrangements, and single nucleotide polymorphism (SNP) changes. One such SNP creates a single amino acid variation in the DNA binding domain of PhoP, the response regulator in the PhoP/PhoQ two-component system. Y. pseudotuberculosis and the basal human-avirulent strains of Y. pestis harbor glycines at position 215 of PhoP, whereas the modern human-virulent strains (e.g., KIM and CO92) harbor serines at this residue. Since PhoP plays multiple roles in the adaptation of Y. pestis to stressful host conditions, we tested whether this amino acid substitution affects PhoP activity or the ability of Y. pestis to survive in host environments. Compared to the parental KIM6+ strain carrying the modern allele of phoP (phoP-S215), a derivative carrying the basal allele (phoP-G215) exhibited slightly defective growth under a low-Mg2+ condition and decreased transcription of a PhoP target gene, ugd, as well as an ∼8-fold increase in the susceptibility to the antimicrobial peptide polymyxin B. The phoP-G215 strain showed no apparent defect in flea colonization, although a phoP-null mutant showed decreased flea infectivity in competition experiments. Our results suggest that the amino acid variation at position 215 of PhoP causes subtle changes in the PhoP activity and raise the possibility that the change in this residue have contributed to the evolution of increased virulence in Y. pestisIMPORTANCEY. pestis acquired a single nucleotide polymorphism (SNP) in phoP when the highly human-virulent strains diverged from less virulent basal strains, resulting in an amino acid substitution in the DNA binding domain of the PhoP response regulator. We show that Y. pestis carrying the modern phoP allele has an increased ability to induce the PhoP-regulated ugd gene and resist antimicrobial peptides compared to an isogenic strain carrying the basal allele. Given the important roles PhoP plays in host adaptation, the results raise an intriguing possibility that this amino acid substitution contributed to the evolution of increased virulence in Y. pestis Additionally, we present the first evidence that phoP confers a survival fitness advantage to Y. pestis inside the flea midgut.

The Cat Flea (Ctenocephalides felis) Immune Deficiency Signaling Pathway Regulates Rickettsia typhi Infection.

Rickettsia species are obligate intracellular bacteria with both conserved and lineage-specific strategies for invading and surviving within eukaryotic cells. One variable component of Rickettsia biology involves arthropod vectors: for instance, typhus group rickettsiae are principally vectored by insects (i.e., lice and fleas), whereas spotted fever group rickettsiae are exclusively vectored by ticks. For flea-borne Rickettsia typhi, the etiological agent of murine typhus, research on vertebrate host biology is facilitated using cell lines and animal models. However, due to the lack of any stable flea cell line or a published flea genome sequence, little is known regarding R. typhi biology in flea vectors that, importantly, do not suffer lethality due to R. typhi infection. To address if fleas combat rickettsial infection, we characterized the cat flea (Ctenocephalides felis) innate immune response to R. typhi Initially, we determined that R. typhi infects Drosophila cells and increases antimicrobial peptide (AMP) gene expression, indicating immune pathway activation. While bioinformatics analysis of the C. felis transcriptome identified homologs to all of the Drosophila immune deficiency (IMD) and Toll pathway components, an AMP gene expression profile in Drosophila cells indicated IMD pathway activation upon rickettsial infection. Accordingly, we assessed R. typhi-mediated flea IMD pathway activation in vivo using small interfering RNA (siRNA)-mediated knockdown. Knockdown of Relish and Imd increased R. typhi infection levels, implicating the IMD pathway as a critical regulator of R. typhi burden in C. felis These data suggest that targeting the IMD pathway could minimize the spread of R. typhi, and potentially other human pathogens, vectored by fleas.

The Distribution and Diversity of Bartonella Species in Rodents and Their Ectoparasites across Thailand.

Our study highlights the surveillance of Bartonella species among rodents and their associated ectoparasites (ticks, fleas, lice, and mites) in several regions across Thailand. A total of 619 rodents and 554 pooled ectoparasites (287 mite pools, 62 flea pools, 35 louse pools, and 170 tick pools) were collected from 8 provinces within 4 regions of Thailand. Bandicota indica (279), Rattus rattus (163), and R. exulans (96) were the most prevalent species of rats collected in this study. Real-time PCR assay targeting Bartonella-specific ssrA gene was used for screening and each positive sample was confirmed by PCR using nuoG gene. The prevalence of Bartonella DNA in rodent (around 17%) was recorded in all regions. The highest prevalence of Bartonella species was found in B. savilei and R. rattus with the rate of 35.7% (5/14) and 32.5% (53/163), respectively. High prevalence of Bartonella-positive rodent was also found in B. indica (15.1%, 42/279), and R. norvegicus (12.5%, 5/40). In contrast, the prevalence of Bartonella species in ectoparasites collected from the rats varied significantly according to types of ectoparasites. A high prevalence of Bartonella DNA was found in louse pools (Polyplax spp. and Hoplopleura spp., 57.1%) and flea pools (Xenopsylla cheopis, 25.8%), while a low prevalence was found in pools of mites (Leptotrombidium spp. and Ascoschoengastia spp., 1.7%) and ticks (Haemaphysalis spp., 3.5%). Prevalence of Bartonella DNA in ectoparasites collected from Bartonella-positive rodents (19.4%) was significantly higher comparing to ectoparasites from Bartonella-negative rodents (8.7%). The phylogenetic analysis of 41 gltA sequences of 16 Bartonella isolates from rodent blood and 25 Bartonella-positive ectoparasites revealed a wide range of diversity among Bartonella species with a majority of sequences (61.0%) belonging to Bartonella elizabethae complex (11 rodents, 1 mite pool, and 5 louse pools), while the remaining sequences were identical to B. phoceensis (17.1%, 1 mite pool, 5 louse pools, and 1 tick pool), B. coopersplainensis (19.5%, 5 rodents, 1 louse pool, and 2 tick pools), and one previously unidentified Bartonella species (2.4%, 1 louse pool).

Rickettsia typhi IN RODENTS AND R. felis IN FLEAS IN YUCATÁN AS A POSSIBLE CAUSAL AGENT OF UNDEFINED FEBRILE CASES / Rickettsia typhi y R. felis en roedores y sus pulgas en Yucatán como posible agente causal de casos febriles indefinidos

Rickettsia typhi is the causal agent of murine typhus; a worldwide zoonotic and vector-borne infectious disease, commonly associated with the presence of domestic and wild rodents. Human cases of murine typhus in the state of Yucatán are frequent. However, there is no evidence of the presence of Rickettsia typhi in mammals or vectors in Yucatán. The presence of Rickettsia in rodents and their ectoparasites was evaluated in a small municipality of Yucatán using the conventional polymerase chain reaction technique and sequencing. The study only identified the presence of Rickettsia typhi in blood samples obtained from Rattus rattus and it reported, for the first time, the presence of R. felis in the flea Polygenis odiosus collected from Ototylomys phyllotis rodent. Additionally, Rickettsia felis was detected in the ectoparasite Ctenocephalides felis fleas parasitizing the wild rodent Peromyscus yucatanicus. This study’s results contributed to a better knowledge of Rickettsia epidemiology in Yucatán.

Rickettsia typhi es el agente causal del tifo murino; una enfermedad zoonótica transmitida por vector mundialmente distribuida, comúnmente asociada con la presencia de roedores domésticos y silvestres. Los casos humanos de tifo murino en el Estado de Yucatán son frecuentes. Sin embargo, no existe evidencia de la presencia de Rickettsia typhi en mamíferos o vectores en Yucatán. En la búsqueda de vectores y reservorios de Rickettsia typhi, evaluamos la presencia de bacterias del género Rickettsia en roedores y sus ectoparásitos de un pequeño municipio del estado de Yucatán por medio de técnicas de PCR convencional y secuenciación de ADN. Se identificó la presencia de Rickettsia typhi en muestras de sangre obtenidas de Rattus rattus y reportamos por primera vez la presencia de Rickettsia felis en la pulga Polygenis odiosus colectado de Ototylomys phyllotis. Complementariamente, Rickettsia felis fue detectado en la pulga Ctenocephalides felis parasitando al roedor Peromyscus yucatanicus. No se identificó especie de Rickettsia en las muestras de sangre de O. phyllotis y P. yucatanicus analizados. Nuestros resultados contribuyen también en el conocimiento de ciclo de vida biológico del género Rickettsia.

Dermal neutrophil, macrophage and dendritic cell responses to Yersinia pestis transmitted by fleas.

Yersinia pestis, the causative agent of plague, is typically transmitted by the bite of an infected flea. Many aspects of mammalian innate immune response early after Y. pestis infection remain poorly understood. A previous study by our lab showed that neutrophils are the most prominent cell type recruited to the injection site after intradermal needle inoculation of Y. pestis, suggesting that neutrophil interactions with Y. pestis may be important in bubonic plague pathogenesis. In the present study, we developed new tools allowing for intravital microscopy of Y. pestis in the dermis of an infected mouse after transmission by its natural route of infection, the bite of an infected flea. We found that uninfected flea bites typically induced minimal neutrophil recruitment. The magnitude of neutrophil response to flea-transmitted Y. pestis varied considerably and appeared to correspond to the number of bacteria deposited at the bite site. Macrophages migrated towards flea bite sites and interacted with small numbers of flea-transmitted bacteria. Consistent with a previous study, we observed minimal interaction between Y. pestis and dendritic cells; however, dendritic cells did consistently migrate towards flea bite sites containing Y. pestis. Interestingly, we often recovered viable Y. pestis from the draining lymph node (dLN) 1 h after flea feeding, indicating that the migration of bacteria from the dermis to the dLN may be more rapid than previously reported. Overall, the innate cellular host responses to flea-transmitted Y. pestis differed from and were more variable than responses to needle-inoculated bacteria. This work highlights the importance of studying the interactions between fleas, Y. pestis and the mammalian host to gain a better understanding of the early events in plague pathogenesis.

Managing iron supply during the infection cycle of a flea borne pathogen, Bartonella henselae.

Bartonella are hemotropic bacteria responsible for emerging zoonoses. Most Bartonella species appear to share a natural cycle that involves an arthropod transmission, followed by exploitation of a mammalian host in which they cause long-lasting intra-erythrocytic bacteremia. Persistence in erythrocytes is considered an adaptation to transmission by bloodsucking arthropod vectors and a strategy to obtain heme required for Bartonella growth. Bartonella genomes do not encode for siderophore biosynthesis or a complete iron Fe(3+) transport system. Only genes, sharing strong homology with all components of a Fe(2+) transport system, are present in Bartonella genomes. Also, Bartonella genomes encode for a complete heme transport system. Bartonella must face various environments in their hosts and vectors. In mammals, free heme and iron are rare and oxygen concentration is low. In arthropod vectors, toxic heme levels are found in the gut where oxygen concentration is high. Bartonella genomes encode for 3-5 heme-binding proteins. In Bartonella henselae heme-binding proteins were shown to be involved in heme uptake process, oxidative stress response, and survival inside endothelial cells and in the flea. In this report, we discuss the use of the heme uptake and storage system of B. henselae during its infection cycle. Also, we establish a comparison with the iron and heme uptake systems of Yersinia pestis used during its infection cycle.

[Role of endoparasites of fleas of wild rodents in plague enzootia].

Laboratory studies of fleas for gregarines have established that it is the latter inhabiting the intestine and stomach of the fleas of wild rodents which are of much interest as protozoa, in whose organism, parasitic species of bacteria can survive. Penetration of plague bacteria into the endoplasm ofgregarines and their possible survival in the cysts may create an additional component in the chain of an epizootic process, which ensures its function, without involving the rodents at the nesting biocenotic level following the pattern: flea imagoes - nesting litter infected with gregarine spores, cysts - flea larvae - flea imagoes infected with cysts, with the plague pathogen emerging into a rodent population through the imago of blocked fleas.

Plague: history and contemporary analysis.

Plague has caused ravaging outbreaks, including the Justinian plague and the "black death" in the Middle Ages. The causative agents of these outbreaks have been confirmed using modern molecular tests. The vector of plague during pandemics remains the subject of controversy. Nowadays, plague must be suspected in all areas where plague is endemic in rodents when patients present with adenitis or with pneumonia with a bloody expectorate. Diagnosis is more difficult in the situation of the reemergence of plague, as in Algeria for example, told by the first physician involved in that outbreak (NM). When in doubt, it is preferable to prescribe treatment with doxycycline while waiting for the test results because of the risk of fatality in individuals with plague. The typical bubo is a type of adenitis that is painful, red and nonfluctuating. The diagnosis is simple when microbiological analysis is conducted. Plague is a likely diagnosis when one sees gram-negative bacilli in lymph node aspirate or biopsy samples. Yersinia pestis grows very easily in blood cultures and is easy to identify by biochemical tests and MALDI-TOF mass spectrometry. Pneumonic plague and septicemic plague without adenitis are difficult to diagnose, and these diagnoses are often made by chance or retrospectively when cases are not part of an epidemic or related to another specific epidemiologic context. The treatment of plague must be based on gentamicin or doxycycline. Treatment with one of these antibiotics must be started as soon as plague is suspected. Analysis of past plague epidemics by using modern laboratory tools illustrated the value of epidemic buboes for the clinical diagnosis of plague; and brought new concepts regarding its transmission by human ectoparasites.

Evidence of antibodies to spotted fever group rickettsiae in small mammals and quail from Mississippi.

Rickettsia parkeri is a recently recognized human pathogen primarily associated with the Gulf Coast tick Amblyomma maculatum, with immature stages of this tick reported from wild vertebrates. To better understand the role of vertebrates in the natural history of this bacterium, we evaluated small mammals and ground-dwelling birds for evidence of infection with R. parkeri or exposure to the organism. We sampled small mammals (n=39) and passerines (n=47) in both north-central and southeast Mississippi, while northern bobwhite (Colinus virginianus) samples (n=31) were obtained from farms in central Mississippi. Blood from all sampled animals was tested using polymerase chain reaction (PCR) for spotted fever group rickettsiae (SFGR), and for antibodies to SFGR using R. parkeri antigen. Ectoparasite samples were removed from animals and included mites, lice, fleas, and immature ticks. Of 39 small mammal samples collected, 7 were positive for antibodies to SFGR; none tested positive by PCR for DNA of SFGR. Of 47 passerine blood samples collected, none were positive for DNA of SFGR by PCR, nor did any show serological evidence of exposure. Finally, none of 31 northern bobwhite samples tested were positive for SFGR DNA, while 7 were seropositive for rickettsial antibodies. Detection of seropositive rodents and quail suggests a role for these host species in the natural history of SFGR, possibly including R. parkeri, but the extent of their role has not yet been elucidated.

Multispacer typing of Rickettsia isolates from humans and ticks in Tunisia revealing new genotypes.

BACKGROUND: Rickettsioses are important remerging vector born infections. In Tunisia, many species have been described in humans and vectors. Genotyping is important for tracking pathogen movement between hosts and vectors. In this study, we characterized Rickettsia species detected in patients and vectors using multispacer typing (MST), proposed by Founier et al. and based on three intergenic spacers (dksA-xerC, rmpE- tRNA(fMet), mppA-pruC) sequencing. METHODS: Our study included 25 patients hospitalized during 2009. Ticks and fleas were collected in the vicinity of confirmed cases. Serology was performed on serum samples by microimmunofluorescence using Rickettsia conorii and Rickettsia typhi antigens. To detect and identify Rickettsia species, PCR targeting ompA, ompB and gltA genes followed by sequencing was performed on 18 obtained skin biopsies and on all collected vectors. Rickettsia positive samples were further characterized using primers targeting three intergenic spacers (dksA-xerC, rmpE- tRNA(fMet) and mppA-purC). RESULTS: A rickettsial infection was confirmed in 15 cases (60%). Serology was positive in 13 cases (52%). PCR detected Rickettsia DNA in four biopsies (16%) allowing the identification of R. conorii subsp israelensis in three cases and R. conorii subsp conorii in one case. Among 380 collected ticks, nine presented positive PCR (2.4%) allowing the identification of six R. conorii subsp israelensis, two R. massiliae and one R. conorii subsp conorii. Among 322 collected fleas, only one was positive for R. felis. R. conorii subsp israelensis strains detected in humans and vectors clustered together and showed a new MST genotype. Similarly, R. conorii subsp conorii strains detected in a skin biopsy and a tick were genetically related and presented a new MST genotype. CONCLUSIONS: New Rickettsia spotted fever strain genotypes were found in Tunisia. Isolates detected in humans and vectors were genetically homogenous despite location differences in their original isolation suggesting epidemiologic circulation of these strains.

Role of a new intimin/invasin-like protein in Yersinia pestis virulence.

A comprehensive TnphoA mutant library was constructed in Yersinia pestis KIM6 to identify surface proteins involved in Y. pestis host cell invasion and bacterial virulence. Insertion site analysis of the library repeatedly identified a 9,042-bp chromosomal gene (YPO3944), intimin/invasin-like protein (Ilp), similar to the Gram-negative intimin/invasin family of surface proteins. Deletion mutants of ilp were generated in Y. pestis strains KIM5(pCD1(+)) Pgm(-) (pigmentation negative)/, KIM6(pCD1(-)) Pgm(+), and CO92. Comparative analyses were done with the deletions and the parental wild type for bacterial adhesion to and internalization by HEp-2 cells in vitro, infectivity and maintenance in the flea vector, and lethality in murine models of systemic and pneumonic plague. Deletion of ilp had no effect on bacterial blockage of flea blood feeding or colonization. The Y. pestis KIM5 Δilp strain had reduced adhesion to and internalization by HEp-2 cells compared to the parental wild-type strain (P < 0.05). Following intravenous challenge with Y. pestis KIM5 Δilp, mice had a delayed time to death and reduced dissemination to the lungs, livers, and kidneys as monitored by in vivo imaging using a lux reporter system (in vivo imaging system [IVIS]) and bacterial counts. Intranasal challenge in mice with Y. pestis CO92 Δilp had a 55-fold increase in the 50% lethal dose ([LD(50)] 1.64 × 10(4) CFU) compared to the parental wild-type strain LD(50) (2.98 × 10(2) CFU). These findings identified Ilp as a novel virulence factor of Y. pestis.

Detección de Rickettsia spp. en ectoparásitos de animales domésticos y silvestres de la Reserva Natural Privada Cerro Chucantí y comunidades aledañas, Panamá, 2007-2010 / Detection of Rickettsia in ectoparasites of wild and domestic mammals from the Cerro Chucanti private reserve and from neighboring towns, Panamá, 2007-2010

Introducción. Los ectoparásitos son los principales vectores de rickettsiosis. En Panamá se tienen escasos datos sobre los artrópodos que pudieran considerarse vectores o reservorios. Objetivos. Presentar datos sobre la presencia de Rickettsia spp. en ectoparásitos de fauna silvestre y animales domésticos en la Reserva Natural Privada Cerro Chucantí y poblados vecinos. Materiales y métodos. Se revisaron 9 personas, 95 mamíferos domésticos y 48 silvestres. Los animales domésticos se examinaron con anuencia del propietario, mientras que la fauna silvestre se capturó con trampas Sherman y Tomahawk. Se extrajeron 21 especies de ectoparásitos: pulgas, piojos, garrapatas y otros ácaros, los cuales se preservaron en etanol al 95 %. Se extrajo material genético de garrapatas y pulgas para ser analizado por técnicas moleculares en la detección de Rickettsia spp. Resultados. Se practicaron 425 reacciones de PCR, de las cuales, 270 resultaron negativas y 155 positivas. De las positivas, 86 amplificaron para el gen gltA (55 % de las positivas); de estos también amplificaron 41 (26 %) para ompA. Se encontró material genético de Rickettsia amblyommii, en garrapatas de caballos (Amblyomma cajennense, Dermacentor nitens), de perros (Rhipicephalus sanguineus) y ninfas de Amblyomma recolectadas en el bosque. Además, se detectó ADN de R. felis en pulgas Ctenocephalides felis de perros. Conclusiones. Se pudo detectar la presencia de R. amblyommii y R. felis en garrapatas y pulgas de animales domésticos de los poblados cercanos a Cerro Chucantí, aun cuando no se pudo encontrar material genético de Rickettsia en ectoparásitos de la fauna silvestre.

Introduction. Ectoparasites are the main vectors of rickettsiosis. In Panama, however, limited data are available concerning the arthropod species that serve as vectors or reservoirs. Objectives. Data are presented concerning the presence of Rickettsia in ectoparasites of wildlife and domestic animals in the Cerro Chucantí private nature reserve and in neighboring villages. Materials and methods. Nine humans, 95 domestic mammals and 48 wild mammals were examined. Twenty-one species of ectoparasites were obtained, including fleas, lice, ticks and mites. These were preserved in 95% ethanol. Later, the DNA was extracted from the ticks and fleas and analyzed by molecular techniques to detect presence of Rickettsia. Results. Of a total of 425 PCR reactions, 270 were positive for Rickettsia and 155 negative. Among the positive samples, 86 PCR amplified for the gltA gene (55% of positives) and 41 of these also amplified the ompA gene. DNA of Rickettsiaamblyommii was found in horses ticks (Amblyomma cajennense, Dermacentor nitens), dogs ticks (Rhipicephalus sanguineus) and free living nymphs in the forest. Additionally, DNA of R. felis was found in fleas from dogs Ctenocephalides felis. Conclusions. The presence of R. amblyommii and R. felis was detected in ticks and fleas of domestic animals in villages near Cerro Chucanti; however no Rickettsia DNA was found in ectoparasites of non-domestic wildlife.

Infecção por Yersinia pestis, no Estado da Bahia: controle efetivo ou silêncio epidemiológico? / Yersinia pestis infection in the State of Bahia: effective control or epidemiological silence?

INTRODUÇÃO: A peste, doença infectocontagiosa milenar, continua sendo considerada da maior importância do ponto de vista epidemiológico devido ao alto potencial epidêmico, estando inclusive sujeita ao Regulamento Sanitário Internacional. Apesar da ausência de casos humanos da doença no Brasil, seu agente etiológico, a bactéria Yersinia pestis, permanece firmemente arraigado em seus focos naturais. A ocorrência de sorologia positiva em carnívoros domésticos de regiões pestígenas da Bahia, nos últimos anos, objetivou a realização deste estudo, que se propõe a verificar a existência de circulação do agente no estado, tendo em vista que fatores condicionantes para a doença são mantidos, oferecendo riscos à população. MÉTODOS: Trata-se de um estudo para verificação da presença de infecção por Y. pestis através do inquérito de soroprevalência em humanos, cães e roedores; e pesquisa da bactéria em roedores e pulgas. Utilizou-se de questionário estruturado para avaliação da associação existente entre fatores ambientais, sócioeconômicos e biológicos e a soroprevalência da infecção em humanos. RESULTADOS: Os 630 soros examinados (88 de humanos, 480 de cães, 62 de roedores) apresentaram-se não reagentes para peste e as análises bacteriológicas realizadas em 14 roedores e dois lotes de pulgas não identificaram a bactéria. CONCLUSÕES: Os resultados não configuram erradicação da doença no estado, pois sua natureza cíclica indica que pode passar longos períodos silente e depois ressurgir acometendo um grande número de pessoas. Portanto, a manutenção da vigilância ativa e permanente se faz necessária para a detecção precoce da doença e desenvolvimento oportuno das medidas de controle pertinentes.

INTRODUCTION: From an epidemiological point of view, the plague is still being considered of great importance, because of its high epidemic potential. Despite the absence of cases of human plague in Brazil, its etiologic agent, the bacteria Yersinia pestis, is still deep rooted in its natural environment. The occurrence of positive serology for plague in domestic carnivores in plague areas in Bahia in the past few years implies the need for a more rigorous evaluation in order to verify whether the bacillus of the plague is still active in these areas. METHODS: In this study, the presence of infection caused by Y. pestis was analyzed by seroprevalence tests on humans, dogs and rodents and by the detection of the bacteria in rodents and fleas. A structured questionnaire was used to analyze the association between environmental, socioeconomic and biological factors and seroprevalence in humans. RESULTS: Of the 630 serum samples examined (88 from humans, 480 from dogs and 62 from rodents), all were nonreactive for plague and bacteriological analyses performed on 14 rodents and 2 flea lots showed no signs of the bacteria. CONCLUSIONS: These results cannot confirm the eradication of the disease in the entire State, since the cyclic nature of the plague indicates that it can go silent for long periods and then resurge, affecting large numbers of people. Thus, maintenance of active, permanent surveillance is required for early detection and the development of adequate control measures.

Prevalence of Bartonella species DNA and antibodies in cats (Felis catus) submitted to a spay/neuter program in Rio de Janeiro, Brazil.

The prevalence of Bartonella species DNA and antibodies for Bartonella henselae were studied in 40 clinically healthy cats (Felis catus, Linnaeus 1758) submitted to a spay/neuter program in Rio de Janeiro, Brazil. Additionally, the prevalence of Bartonella species DNA was investigated in the fleas found parasitizing the subject cats. For this purpose, blood samples were obtained from all cats, and DNA extraction was performed on the blood, and blood clotted samples, as well as on pools of fleas obtained from them. Antibodies for B henselae were detected on serum samples. Bartonella species DNA was detected in 17 cats, whereas serum reactivity for B henselae was found in 19. A total of 20 cats were flea-infested and nine of these 20 had Bartonella species DNA in their blood. In four of the 20 flea-infested cats, Bartonella species DNA was detected in the fleas obtained from those cats, but only one of these four cats had Bartonella species DNA in its blood.