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Background: Spotted Fever Group (SFG) Rickettsiae can cause febrile diseases with or without rash in humans worldwide. In Germany only limited data are available about their medical significance. Serological screening tests for antibodies against rickettsiae usually only distinguish between SFG and Typhus Group (TG) Rickettsiae due to the strong cross reactivities within the groups. Seroprevalence rates in dogs, as possible sentinels for tick-borne diseases, could serve as an indicator for the distribution of different Rickettsia species., Methods: In this study, a micro-immunofluorescence assay (micro-IFA) was established for detection and differentiation of antibodies against five Rickettsia species in dogs (R. helvetica, R. raoultii, R. slovaca, R. monacensis and R. felis). Dogs that never left Germany (n = 605) previously investigated with an SFG-ELISA were included in this study and screened at a 1:128 dilution. Endpoint titres of fifty randomly selected seropositive samples of each of the five investigated regions in Germany were determined in order to allow a differentiation of the causative Rickettsia species. Sensitivity and specificity of the micro-IFA were compared with ELISA results of the previous study., Results: A total of 93.9% of the dogs were positive for antibodies of the SFG Rickettsiae at the screening titer of 1:128. Differentiation of SFG Rickettsiae with the micro-IFA was possible in 70.4%, but in 29.6% of the cases the detected antibodies were not differentiable. Considering a clear differentiation by a twofold titre difference between observed reactions, the seroprevalence rates were 66.0% for R. helvetica, 2.8% for R. raoultii, 1.6% for R. slovaca, but no serological reaction could be clearly attributed to R. monacensis or R. felis. No statistically significant regional differences were found for R. helvetica, R. slovaca and R. raoultii comparing the five regions of Germany. Comparison of micro-IFA with ELISA revealed a sensitivity of 82.0% and a specificity of 83.8% for the Rickettsia SFG ELISA., Conclusions: The micro-IFA is a useful serological tool to differentiate antibodies against different Rickettsia species in dogs. Seroprevalence rates in dogs correspond to the prevalence rates and distribution of Rickettsia-carrying tick species.

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 (prevalence 1.9%) and R. felis (prevalence 24.8%) DNA were detected in rat fleas (Xenopsylla cheopis) collected from mice on Oahu Island, Hawaii. The low prevalence of R. typhi on Oahu suggests that R. felis may be a more common cause of rickettsiosis than R. typhi in Hawaii.

Rickettsia typhi and R. felis are flea-transmitted human pathogenic rickettsial species. To investigate the distributional dynamics of these rickettsiae we designed a micro-immunofluorescence assay (MIF) using species-specific monoclonal antibodies (MAbs) applied to flea cryosections. Our assay was performed in less than 3 h and its applicability was demonstrated by the detection of R. typhi in 50 artificially infected human body lice but in none of 50 uninfected lice. With MIF, we identified 31 positive among 32 fleas proven with PCR to be naturally infected with R. felis; and 7 positive among 32 fleas proven with PCR to be naturally infected with R. typhi. No cross-detection was observed with both MAbs. Fresh R. felis-infected fleas were significantly more MIF-positive than long conserved R. typhi-infected fleas (31/32 vs. 7/32, P < 0.01). This discrepancy may be linked to degradation of antigens by long-term freezing. For R. typhi-infected fleas, our assay was significantly more efficient when applied to fleas in early stages of infection (less than 15 days) by comparison with fleas frozen more than 20 days after infection (7/15 vs. 0/17, P = 0.01). This difference may be related to an antigenic modification caused by selection pressure in the vector and host process. The sensitivity of the described method did not exceed 47% (7/15) for R. typhi but, in contrast, was 97% for R. felis. Thus, our method appears to be useful for surveillance in R. felis infections, but requires further studies for the detection of R. typhi.

Rickettsia typhi and R. felis, 2 closely related rickettsial species, often have been identified in cat fleas, Ctenocephalides felis (Bouché) from the same geographical location. However, no fleas have been found to be naturally infected concurrently with both rickettsial species. To examine whether one rickettsial species can develop simultaneously with another species in the same flea host, cat fleas, naturally infected with R. felis, were allowed to feed on blood containing R. typhi (Ethiopian strain, 10(7) PFU/ml). Experimental controls consisted of uninfected cat fleas, fleas infected with only R. typhi; and fleas naturally infected with only R. felis. After 9 d at 28 degrees C, the fleas were examined by PCR amplification and subsequent restriction digest analysis and dot blot hybridization of PCR products. Results from these studies demonstrated that R. felis and R. typhi are capable of co-existing in the same flea host. Subsequent isolation of R. typhi from dually infected fleas by tissue culture indicated that R. typhi was viable and capable of being maintained in fleas naturally infected with R. felis. As more studies confirm the presence of R. felis and R. typhi in domestic pets and peridomestic vertebrates in urban areas, the ability of the individual cat fleas, which live on these animals, to support both rickettsial species could be an epidemiologically important consideration.

Rickettsia species are bacteria that may cause multiple diseases in animals and humans, via transmission through multiple arthropod vectors. Routine surveillance of Rickettsia spp. within vectors is critical to determine their presence and risk to mammalian hosts within human populations. Therefore, to better characterize the circulating Rickettsia species in an understudied region we targeted pet dogs to survey. Ticks were collected from pet dogs in three populations of the Yucatan where we tested for the presence of Rickettsia spp. by PCR in metagenomic DNA. In these ticks removed from pet dogs we detected Rickettsia amblyommatis and Rickettsia bellii in Amblyomma auriculatum, Amblyomma ovale and Amblyomma mixtum ticks obtained in a rural community in the Mexican state of Yucatan. This is the first report detecting both species for this state in Mexico, underpinning the importance of more routine surveillance., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Murine typhus, caused by Rickettsia typhi , is re-emerging in many parts of the world. The disease is also called endemic typhus to differentiate from epidemic typhus (caused by Rickettsia prowazekii ), and sometimes also named flea-borne typhus. Occasionally, literature sources will include Rickettsia felis as a causative agent of flea-borne typhus, but illnesses caused by R. felis are actually flea-borne spotted fever. Murine typhus occurs in warm, coastal areas worldwide. In the United States, most cases are reported from California, Texas, and Hawaii. Murine typhus is usually a self-limited febrile illness but about one-quarter of patients suffer organ complications. The disease is only infrequently fatal. Regarding disease ecology, the historical paradigm is that rats ( Rattus rattus and R. norvegicus ) are reservoirs of R. typhi worldwide, with rat fleas ( Xenopsylla cheopis ) as primary vectors. More recently, researchers have proposed an alternative suburban murine typhus transmission cycle involving opossums, cat fleas, cats, and dogs in Texas, California, and rural Mexico. Because cat fleas feed on a variety of mammals, there may be other avenues for R. typhi transmission, including stray or feral cats bringing cat fleas and other infected fleas into proximity with humans and possible aerosolization of infected flea feces. Additional fleas, ticks, lice, and mites may play a role in various areas throughout the world, but a striking lack of fundamental research on this topic makes drawing conclusions difficult. This review provides an overview of the history, epidemiology, diagnosis, and treatment of murine typhus, with special emphasis on its disease ecology.

Flea-borne diseases are endemic in Texas, U.S.A., with an increasing incidence of flea-borne typhus and cat scratch disease. Knowledge of flea natural history could provide information to protect public health, yet many knowledge gaps remain outside of plague-endemic regions. Our objective was to characterize seasonal activity patterns of fleas on common mammalian wildlife species and test fleas and wildlife for Rickettsia and Bartonella pathogens. We performed one year of monthly trapping for rodents and medium-sized mammals in a national forest with high recreational use and urban encroachment in East Texas. From 90 mammal captures representing seven species, 101 fleas were collected representing Polygenis spp., Ctenocephalides felis , and Orchopeas species. Virginia opossums ( Didelphis virginianus ) hosted 99% of the collected fleas (100 fleas) and a single flea was on an eastern woodrat ( Neotoma floridana ). Flea infestation prevalence of opossums was 79% (23/29). Mean flea abundance was 4.39 fleas, with intensity peaking in spring. One cat flea removed from an opossum was positive for Bartonella henselae . Furthermore, we identified tissue or blood of four raccoons ( Procyon lotor ) and one golden mouse ( Ochrotomys nuttalli ) positive for Rickettsia amblyommatis . These findings provide an ecological basis for the maintenance of vectors and pathogens from sylvatic settings.

Vector-borne diseases account for nearly 20% of all globally recognised infectious diseases. Within the spectrum of flea-borne pathogens, Bartonella and Rickettsia bacteria are prominent, contributing to the emergence and resurgence of diseases on a global scale. This study investigates the presence of species of Bartonella and Rickettsia harboured by fleas collected from wild rodents in northwestern Argentina (NWA). A total of 28 fleas from three genera and seven species were assessed. DNA of Bartonella and Rickettsia spp. was found in 12 fleas (42.8%). Phylogenetic analysis of concatenated sequences of gltA and rpoB genes showed the presence of Bartonella quintana in eight fleas of two species, Craneopsylla minerva minerva and Polygenis acodontis. Phylogenetic analysis of concatenated sequences of gltA, ompA and ompB genes identified Rickettsia felis in ten fleas of five species, C. m. minerva, P. acodontis, Polygenis bohlsi bohlsi, Polygenis byturus and Tiamastus palpalis. These bacterial species mark the first report in all flea species studied. This study represents the first survey of flea-borne bacteria for NWA. The results provide information to address strategies for the control and prevention of bartonellosis and rickettsiosis that could have an impact on public health in one of the geographical areas of Argentina with the highest incidence of infections transmitted to humans by ectoparasites., (© 2024. The Author(s).)

Background: In urban Thailand, arboviral infections dominate diagnoses of acute undifferentiated fevers (AUFs) owing to their well-defined epidemiology and characteristic clinical presentations. However, rickettsial diseases, also endemic in this setting, remain under-recognized owing to challenges in early detection. Objective: This study aimed to identify potential rickettsial infections among patients with AUF in Bangkok and vicinity utilizing leftover nucleic acid extracted from serum samples from patients initially suspected of but negative for arbovirus infections. Materials and Methods: A total of 609 nucleic acid samples were screened for rickettsial bacteria using real-time PCR, targeting the 17-kDa common antigen gene of Rickettsia spp. and the 47-kDa gene of Orientia tsutsugamushi . Results: Nine samples were positive for Rickettsia spp. and two were positive for O. tsutsugamushi . DNA sequence and phylogenetic analyses based on partial 17-kDa antigen and citrate synthase ( gltA ) genes identified the Rickettsia -positive samples as R. typhi in eight cases and R. felis in one case. Analysis of the 56-kDa type-specific antigen gene identified the two O. tsutsugamushi isolates as Gilliam-related genotypes. Although rickettsial diseases typically present with mild symptoms, two patients with R. typhi infection (murine typhus) developed respiratory distress syndrome, highlighting the potential for rare but serious complications. Conclusion: This study underscores the critical importance of differential diagnosis and prompt, effective intervention to prevent complications in suspected cases.

Introduction: Rickettsioses are zoonotic diseases transmitted by arthropods acting as vectors and reservoirs. Disease symptoms are nonspecific and, therefore, their clinical diagnosis is difficult. Indirect immunofluorescence (IFA) is the gold standard assay for diagnosis. The interest for conducting studies on these pathologies has resurfaced in Colombia since 2001; besides, previous studies have evidenced cases of rickettsiosis in the north of the department of Caldas. Objective: To establish the frequency of antibodies and seroconversion against Rickettsia spp. In patients consulting health institutions in Caldas, Colombia, from 2016 to 2019. Materials and methods: We conducted a quantitative, observational, and descriptive study on a non-probabilistic sample of 175 patients with symptoms compatible with rickettsiosis who consulted in different municipalities of Caldas, Colombia; IFA was performed to detect antibodies in the acute and convalescent phases against Rickettsia rickettsii, Rickettsia typhi, and Rickettsia felis. Results: The average age of the patients was 31 years. The municipalities with the highest proportion of seropositive cases were Belalcázar, Chinchiná, Filadelfia, La Dorada, La Merced, and Manizales; 66% of patients owned pets and 12% reported arthropod bites. The most frequent signs and symptoms were headache (69.7%), arthromyalgia (60%), and fever (58.2%). IgG seroprevalence was 60% for R. rickettsii, 47.9% for R. typhi, and, and 24% for R. felis. Eight patients presented seroconversion. Conclusion: We found evidence of the circulation of Rickettsia species from the spotted fever group and the typhus group associated with human cases in Caldas.

Genetic diversity is lost in small and isolated populations, affecting many globally declining species. Interspecific admixture events can increase genetic variation in the recipient species' gene pool, but empirical examples of species-wide restoration of genetic diversity by admixture are lacking. Here we present multi-fold coverage genomic data from three ancient Iberian lynx (Lynx pardinus) approximately 2,000-4,000 years old and show a continuous or recurrent process of interspecies admixture with the Eurasian lynx (Lynx lynx) that increased modern Iberian lynx genetic diversity above that occurring millennia ago despite its recent demographic decline. Our results add to the accumulating evidence for natural admixture and introgression among closely related species and show that this can result in an increase of species-wide genetic diversity in highly genetically eroded species. The strict avoidance of interspecific sources in current genetic restoration measures needs to be carefully reconsidered, particularly in cases where no conspecific source population exists., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Rickettsia parkeri belongs to the spotted fever group (SFG) of the Rickettsia genus. This bacterium causes mild rickettsiosis in humans and is mainly transmitted by Amblyomma ticks. Its medical importance is emerging in the Americas, including Mexico. Synanthropic rodents and domiciled dogs participate as accidental hosts in epidemiological cycles of Rickettsia of the SFG. The aim is to report the presence of R. parkeri in synanthropic rodents and domiciled dogs from a rural community of Yucatán, Mexico. Rodents were captured, and plasma samples were taken from dogs in 48 households from Ucú, Yucatán, Mexico. A spleen sample (rodents) and plasma (dogs) were used in the propagation of Rickettsia on Vero cells. These infected cells were used in the extraction of genomic DNA. Rickettsia DNA was identified using a semi-nested PCR (snPCR); some products were sent for sequencing. The recovered sequences were analysed with bioinformatics programs, and a phylogenetic tree was built to determine the Rickettsia species. One hundred animals were sampled: 36 synanthropic rodents and 64 dogs. The snPCR evidenced the presence of Rickettsia DNA in 10 rodents (10/36, 27.8%) and 18 dogs (18/64, 28.1%), which represents a global frequency of 28% (28/100) in this study. The bioinformatics analysis yielded homology to R. parkeri and was demonstrated in the phylogenetic tree. The first evidence of the presence of R. parkeri in synanthropic rodents (Mus musculus) from Mexico is presented; likewise, the participation of domestic dogs in the transmission cycle of this bacterium with potential importance in public health is confirmed., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Antibiotic resistance genes (ARGs) are a growing global threat to public health. Biological laboratory wastewater contains large amounts of free ARGs. It is important to assess the risk of free ARGs from biological laboratories and to find appropriate treatments to control their spread. The fate of plasmids in the environment and the effect of different thermal treatments on their persistence activity were tested. The results showed that untreated resistance plasmids could exist in water for more than 24 h (the special 245 bp fragment). Gel electrophoresis and transformation assays showed that the plasmids boiled for 20 min retained 3.65% ± 0.31% transformation activity of the intact plasmids, while autoclaving for 20 min at 121 °C could effectively degrade the plasmids and that NaCl, bovine serum albumin, and EDTA-2Na affected the degradation efficiency of the plasmids during boiling. In the simulated aquatic system, using 10 6 copy/μL of plasmids after autoclaving, only 10 2 copies/μL of the fragment after only 1-2 h could be detected. By contrast, boiled plasmids for 20 min were still detectable after plunging them into water for 24 h. These findings suggest that untreated and boiled plasmids can remain in the aquatic environment for a certain time resulting in the risk of disseminating ARGs. However, autoclaving is an effective way of degrading waste free resistance plasmids., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

River ecosystems are very important parts of the water cycle and an excellent habitat, food, and drinking water source for many organisms, including humans. Antibiotics are emerging contaminants which can enter rivers from various sources. Several antibiotics and their related antibiotic resistance genes (ARGs) have been detected in these ecosystems by various research programs and could constitute a substantial problem. The presence of antibiotics and other resistance cofactors can boost the development of ARGs in the chromosomes or mobile genetic elements of natural bacteria in rivers. The ARGs in environmental bacteria can also be transferred to clinically important pathogens. However, antibiotics and their resistance genes are both not currently monitored by national or international authorities responsible for controlling the quality of water bodies. For example, they are not included in the contaminant list in the European Water Framework Directive or in the US list of Water-Quality Benchmarks for Contaminants. Although ARGs are naturally present in the environment, very few studies have focused on non-impacted rivers to assess the background ARG levels in rivers, which could provide some useful indications for future environmental regulation and legislation. The present study reviews the antibiotics and associated ARGs most commonly measured and detected in rivers, including the primary analysis tools used for their assessment. In addition, other factors that could enhance antibiotic resistance, such as the effects of chemical mixtures, the effects of climate change, and the potential effects of the coronavirus disease 2019 pandemic, are discussed. Environ Toxicol Chem 2022;41:687-714. © 2022 SETAC., (© 2022 SETAC.)

Background: Rickettsia spp. are human pathogens that cause a number of diseases and are transmitted by arthropods, such as ixodid ticks. Estonia is one of few regions where the distribution area of two medically important tick species, Ixodes persulcatus and I. ricinus, overlaps. The nidicolous rodent-associated Ixodes trianguliceps has also recently been shown to be present in Estonia. Although no data are available on human disease(s) caused by tick-borne Rickettsia spp. in Estonia, the presence of three Rickettsia species in non-nidicolous ticks has been previously reported. The aim of this study was to detect, identify and partially characterize Rickettsia species in nidicolous and non-nidicolous ticks attached to rodents in Estonia., Results: Larvae and nymphs of I. ricinus (n = 1004), I. persulcatus (n = 75) and I. trianguliceps (n = 117), all removed from rodents and shrews caught in different parts of Estonia, were studied for the presence of Rickettsia spp. by nested PCR. Ticks were collected from 314 small animals of five species [Myodes glareolus (bank voles), Apodemus flavicollis (yellow necked mice), A. agrarius (striped field mice), Microtus subterranius (pine voles) and Sorex araneus (common shrews)]. Rickettsial DNA was detected in 8.7% (103/1186) of the studied ticks. In addition to identifying R. helvetica, which had been previously found in questing ticks, we report here the first time that the recently described I. trianguliceps-associated Candidatus Rickettsia uralica has been identified west of the Ural Mountains.

The present work provides a review focusing on contaminants of emerging concern (CECs) in aquatic environment, with an emphasis on their occurrence, monitoring, fate, and risk assessment in the research published in the scientific literature in 2019. Several studies revealed that these organic contaminants were detected in many water bodies and suspect, nontarget, and target screening provided an efficient detection for the co-existing organic substances with complex components. Wastewater resource recovery facilities were concurrently considered as a central source, and several specific chemicals have been found to be used as chemical markers to track the source of CECs in some urban watersheds. Reliable monitoring, reliable fate/toxicity assessment, and effective removal that consider CECs as a heterogeneous group rather than single substances will be the challenges for the research community in the future., (© 2020 Water Environment Federation.)

The aim of the study was to describe the epidemiological factors associated with the risks of recent Rickettsia exposure in inhabitants of a rural population from Yucatán, Mexico. The study included 130 inhabitants from Maxcanú, Yucatán. Blood samples were collected to detect IgM and IgG antibodies against Rickettsia typhi and Rickettsia rickettsii by an indirect immunofluorescence antibody test. Additionally, nested polymerase chain reaction was performed to amplify fragments of the 17kDa and sca5 genes. Previously, an epidemiological questionnaire was applied focused on collecting information on personal and housing exposure variables related to the recent infection with Rickettsia to determine epidemiological associations. Results that exhibited a p-value < .25 were included in a generalized multinomial logistic linear model to determine the variables involved with the risk of contact or Rickettsia infection. In all, 76% (99/130) of the participants presented with immunoglobulin titres against the Rickettsia species evaluated, while rickettsial DNA was detected in 35.4% (46/130) of the participants. The association analysis with the personal exposure variables showed that the productive age group (OR = 0.32; 95% CI = 0.10-1.03) and the elders group (OR = 0.12; 95% CI = 0.01-0.83) were protective factors for recent infection with R. typhi, taking as reference the school group. The presence of a family orchard in the home (OR = 7.56; 95% CI = 1.62-35.29) was a risk factor for recent infection with R. rickettsii. Presumably, the presence of ectoparasites (OR = 2.71; 95% CI = 0.90-8.09) at home was a risk factor for recent infection with both Rickettsia species. Recent infection was demonstrated in inhabitants from Maxcanú, Yucatán. A high seropositive frequency was obtained. The results highlight the importance of the family garden and the presence of ectoparasites in the home as risk factors associated with recent infection with Rickettsia in inhabitants from Maxcanú., (© 2020 Blackwell Verlag GmbH.)

Background: In Algeria, Mediterranean spotted fever (MSF), caused by Rickettsia conorii conorii and transmitted by Rhipicephalus sanguineus, is endemic. However, the documentation of cases is rare due to a lack of laboratory facilities. Our aim was to screen for rickettsioses in patients with fever, rash and a possible inoculation eschar., Materials and Methods: Between 2013 and 2015, patients with a fever and a rash presenting at hospitals in the Tizi-Ouzou region were prospectively included in our study. Sera were screened using Indirect Immunofluorescence Assay (IFA) and qPCR was performed on swab samples from eschars., Results: One hundred and sixty-six patients were included. For 57 patients, MSF due to R. conorii conorii was diagnosed by serology and qPCR on a swab eschar sample. Three patients were diagnosed with murine typhus, a flea borne rickettsiosis. In addition, two patients had a positive serology in IgM for R. felis. For nine patients, serology for rickettsiosis was positive, but the specific rickettsia involved could not be determined. Nine patients had a severe disease with neurological involvement or multi-organ failure., Conclusion: Clinicians should routinely screen for rickettsioses in patients and travellers presenting with a rash upon return from Algeria. Doxycycline treatment must be given promptly because the prognosis can be severe., (Copyright © 2018. Published by Elsevier Ltd.)

Rickettsia felis has been reported to be a cause of fever in sub-Saharan Africa, but this association has been poorly evaluated in Gabon. We assessed the prevalence of this bacterium among children <15 years of age in 4 areas of Gabon; the locations were in urban, semiurban, and rural areas. DNA samples from 410 febrile children and 60 afebrile children were analyzed by quantitative PCR. Overall, the prevalence of R. felis among febrile and afebrile children was 10.2% (42/410 children) and 3.3% (2/60 children), respectively. Prevalence differed among febrile children living in areas that are urban (Franceville, 1.3% [1/77]), semiurban (Koulamoutou, 2.1% [3/141]), and rural (Lastourville, 11.2% [15/134]; Fougamou, 39.7% [23/58]). Furthermore, in a rural area (Fougamou), R. felis was significantly more prevalent in febrile (39.7% [23/58]) than afebrile children (5.0% [1/20]). Additional studies are needed to better understand the pathogenic role of R. felis in this part of the world.

Rickettsia felis ( R. felis ) is a Gram-negative obligate intracellular bacterium with a global presence across various hosts, including mammals, humans, and ectoparasites. Herein, we present a case of R. felis infection in a previously healthy 43-year-old male with a history of Clonorchis sinensis and recent corticosteroid use. The identification of R. felis was accomplished through metagenomic next-generation sequencing (mNGS) of the patient's bronchoalveolar lavage fluid (BALF). This case represents the first documented pulmonary infection caused by R. felis in China, confirmed by mNGS analysis of bronchoalveolar lavage fluid (BALF) and reported from Shenzhen, China., Competing Interests: The authors declare no potential conflicts of interest for the research, authorship, and/or publication of this article., (© 2024 Wen et al.)

Tick-borne pathogens in the genus Rickettsia are the causative agents of severe and potentially fatal spotted fever group (SFG) and typhus group diseases in dogs and humans. Climate, habitat, and land-use changes are impacting vector ranges, with expansions potentially resulting in novel pathogens being introduced into naïve locations. Despite the public health importance of SFG Rickettsia, there are relatively few data on the prevalence and diversity of rickettsial pathogens in sub-Saharan Africa. The aim of this study was to characterize the SFG Rickettsia prevalence and diversity in ixodid ticks (104 Amblyomma spp., 160 Rhipicephalus spp., and one Hyalomma truncatum) collected from domestic dogs in Chad, Africa. Ticks were screened for Rickettsia spp. using a nested PCR targeting the 17-kDa gene. Species identification was through bidirectional Sanger sequencing of the 17-kDa, ompA, ompB, and/or gltA gene targets. A total of 43.3 % (115/265) ticks were positive for Rickettsia spp. and six Rickettsia species were identified: R. africae, R. massiliae, R. conorii, R. felis, R. monacensis and Candidatus Rickettsia muridii. Seven additional samples were positive for Rickettsia of undetermined species. Rickettsia africae, an important zoonotic pathogen, was found in 81 % (79/97) of A. variegatum and 29 % (2/7) of an A. marmoreum complex species, a group that infests a wide range of birds and mammals, including humans. Finally, we detected a high diversity of Rickettsia spp., most of which were zoonotic, in Rh. muhsamae. Collectively these data indicate there is a risk of rickettsiosis in Chad and further studies on ticks and rickettsial pathogens in this region are warranted., (Copyright © 2024. Published by Elsevier GmbH.)

The occurrence of sporadic rickettsial infections has been consistently undervalued and overlooked, primarily owing to a limited emphasis on routine examinations for rickettsioses in clinical practice. At present, the immunofluorescence assay is the prevailing diagnostic method for suspected rickettsioses that enables the detection of specific antibodies against rickettsia in human serum. Herein, we present an exceptional instance of rickettsial infection that was characterized by a rare manifestation of extensive pericardial effusion leading to dyspnea and cardiac tamponade. A diagnosis of chronic fibrosing pericarditis was established based on pericardium tissue obtained through pericardiotomy, and a conclusive metagenomic next-generation sequencing test confirmed the presence of Rickettsia felis infection. The cat flea, scientifically known as Ctenocephalides felis , is the predominant carrier of R. felis . An escalating incidence of human R. felis infections has raised concerns, particularly in light of the burgeoning population of domesticated animals in many contemporary societies., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2024 Tien-Lung Po et al.)

Flea-borne spotted fever and flea-borne (murine) typhus are rickettsioses caused by Rickettsia felis and Rickettsia typhi, respectively, and typically present as undifferentiated febrile illnesses. The relative contribution of these agents to flea-borne rickettsioses in California is unclear. We have developed a duplex reverse transcription real-time polymerase chain reaction (RT-rtPCR) assay targeting R. felis- and R. typhi-specific 23S ribosomal RNA single nucleotide polymorphisms to better understand the respective roles of these agents in causing flea-borne rickettsioses in California. This assay was compared with an established duplex R. felis- and R. typhi-ompB rt-PCR assay and was shown to have 1,000-fold and 10-fold greater analytical sensitivity for the detection of R. felis and R. typhi, respectively. Retrospective testing of clinical specimens with both assays established R. typhi as the major etiologic agent of flea-borne rickettsioses in California.

Bed bugs, common blood-feeding pests, have received limited attention regarding their potential involvement in emerging pathogen transmission. This study aimed to investigate the main vector-borne bacteria within bed bugs collected from Tunisian governorates and to genetically characterize the identified species. Molecular screening was conducted on field-collected bed bug samples, targeting zoonotic vector-borne bacteria from the Anaplasmataceae family, as well as the genera Rickettsia, Ehrlichia, Bartonella, and Borrelia. A total of 119 Cimex lectularius specimens were collected and grouped into 14 pools based on sampling Tunisian sites. Using genus-specific PCR assays, DNA of Rickettsia and Ehrlichia spp. was detected in a single pool. Sequencing and BLAST analysis of the obtained partial ompB and dsb sequences from positive samples revealed 100% similarity with those of Ehrlichia canis and Rickettsia felis available in GenBank. Obtained partial sequences showed phylogenetic similarity to R. felis and E. canis isolates found in dogs and ticks from American and European countries. To the best of our knowledge, this study is the first to investigate bed bugs in Tunisia and to report the worldwide identification of R. felis and E. canis DNA in the common bed bug, C. lectularius. These findings highlight the need for further research to explore the potential role of bed bugs in the epidemiology of these vector-borne bacteria., Competing Interests: Declaration of Competing Interest The authors declared that they have no potential conflicts of interest with respect to the research, authorship and publication of this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Purpose: Flea-borne rickettsioses, collectively referred to as a term for etiological agents Rickettsia felis , Rickettsia typhi , and RFLOs ( R. felis -like organisms), has become a public health concern around the world, specifically in the United States. Due to a shared arthropod vector (the cat flea) and clinical signs, discriminating between Rickettsia species has proven difficult. While the effects of microbial coinfections in the vector can result in antagonistic or synergistic interrelationships, subsequently altering potential human exposure and disease, the impact of bacterial interactions within flea populations remains poorly defined. Methods: In this study, in vitro and in vivo systems were utilized to assess rickettsial interactions in arthropods. Results: Coinfection of both R. felis and R. typhi within a tick-derived cell line indicated that the two species could infect the same cell, but distinct growth kinetics led to reduced R. felis growth over time, regardless of infection order. Sequential flea coinfections revealed the vector could acquire both Rickettsia spp. and sustain coinfection for up to 2 weeks, but rickettsial loads in coinfected fleas and feces were altered during coinfection. Conclusion: Altered rickettsial loads during coinfection suggest R. felis and R. typhi interactions may enhance the transmission potential of either agent. Thus, this study provides a functional foundation to disentangle transmission events propelled by complex interspecies relationships during vector coinfections.

Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Ticks are small and adaptable arachnid ectoparasites and global carriers of various pathogens that threaten both human and animal health. They are present in many parts of China. A total of 858 ticks were collected from various regions and hosts, then subjected to species identification based on morphological and molecular characteristics, as described in the authors' previous study. Eighty-three individual tick samples were selected for screening pathogens based on metagenomic next-generation sequencing (mNGS) and polymerase chain reaction (PCR) assays. The genomic DNA of tick species was extracted, and amplification of the bacterial 16S rRNA gene was carried out from DNA of individual ticks using V3-V4 hypervariable regions, before subjecting to metagenomic analysis. Each tick underwent specific PCR tests for identifying the bacterial species present, including Anaplasma, Ehrlichia, Coxiella, and Rickettsia, and also protozoans such as Babesia, Theileria, and Hepatozoon. Illumina NovaSeq sequencing results revealed that the dominant phylum and family in Rhipicephalus spp. were Bacteroidota and Muribaculaceae, respectively. Alpha diversity patterns varied depending on tick sex (R. linnaei only), species and location, but not on host. Furthermore, bacterial pathogens, including A. marginale (58 %, 29/50), A. platys (6 %, 3/50), E. minasensis (2 %, 1/50), Ehrlichia sp. (10 %, 5/50), T. sinensis (24 %, 12/50), T. orientalis (54 %, 27/50) and Coxiella-like bacteria (CLB) (80 %, 40/50) were detected in R. microplus, while E. canis (33.33 %, 10/30), H. canis (20 %, 6/30) and CLB (100 %, 30/30) were detected in R. linnaei. Also, Anaplasma sp. (33.33 %, 1/3), A. marginale (33.33 %, 1/3), R. felis (33.33 %, 1/3) and CLB (100 %, 3/3) were detected in R. haemaphysaloides. Dual and triple co-infections involving pathogens or CLB were detected in 84.00 % of R. microplus, 66.66 % of R. haemaphysaloides, and 33.00 % of R. linnaei. The report on microbial communities and pathogens, which found from Rhipicephalus spp. in Hainan Island, is an important step towards a better understanding of tick-borne disease transmission. This is the first report in the area on the presence of Anaplasma sp., A. marginale, R. felis and Coxiella, in R. haemaphysaloides., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Subject: Rickettsia is a zoonotic bacterial pathogen transmitted by vectors and has extensive reservoirs in animal and human populations. Rickettsiosis is a public health problem all over the world. However, comprehensive information on the geographical distribution of different Rickettsia species, infection status of reservoirs, vectors, and human cases is lacking in most parts of the world. Therefore, this study aimed to investigate the geographical distribution of different Rickettsia species and their vectors in countries of the WHO-EMRO region., Methods: In this review study, a search was conducted for reports and published studies on Rickettsia species from WHO-EMRO region countries in various databases from 1995 to 2022. Finally, the reported status of human cases, reservoirs, and vectors associated with each species in different countries was documented., Results: Reports of infections related to the detection of Rickettsia species were only available for 15 out of 22 WHO-EMRO member countries. A total of twenty-four Rickettsia species, including R. sibrica, R. lusitaniae, R. africae, R. prowazekii, R. felis, R. typhi, R. rickettsii, R. aeschlimannii, R. conorii, R. massiliae, R. helvetica, R. monacensis, R. rhipicephali, R. bellii, R. asembonensis, R. hoogstraalii, R. andeanae, R. raoultii, R. asiatica, R. slovaca, R. australis, R. barbariae, Candidatus R. amblyommii, and Candidatus R. goldwasserii, were reported from WHO-EMRO member countries. Furthermore, human cases infected with six different Rickettsia species, including R. sibrica, R. prowazekii, R. felis, R. typhi, R. rickettsii, R. aeschlimannii, R. conorii, R. massiliae, and R. helvetica, were reported from these countries., Conclusion: The vast diversity of Rickettsia vectors has contributed to the ongoing discovery of new Rickettsia species. Therefore, further research on the reservoir hosts of Rickettsia infections in the understudied WHO-EMRO region is crucial. This research sheds light on Rickettsia disease's epidemiology and transmission dynamics in this region., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Ticks are obligate ectoparasites and vectors of pathogens affecting health, agriculture, and animal welfare. This study collected ticks from the cattle and questing ticks of 24 Magdalena Medio Antioquia region cattle farms. Genomic DNA was extracted from the specimens (individual or pools) of the 2088 adult ticks collected from cattle and 4667 immature questing ticks collected from pastures. The molecular detection of Babesia, Anaplasma, Coxiella and Rickettsia genera was performed by polymerase chain reaction amplification and subsequent DNA sequencing. In a total of 6755 Rhipicephalus microplus DNA samples, Anaplasma marginale was the most detected with a frequency of 2% (Confidence Interval- CI 1.68-2.36), followed by Babesia bigemina with 0.28% (CI 0.16-0.44), Coxiella spp. with 0.15% (CI 0.07-0.27), and Rickettsia spp. with 0.13% (CI 0.06-0.25). Molecular analysis of the DNA sequences obtained from the tick samples revealed the presence of Coxiella-like endosymbiont and R. felis. These results demonstrated the diversity of microorganisms present in R. microplus ticks predominantly associated with cattle and questing ticks from livestock agroecosystems, suggesting their role as reservoirs and potential biological vectors of these microorganisms on the studied sites. Also, it emphasizes the need to combine acarological surveillance with clinical diagnoses and control strategies on regional and national levels., Competing Interests: Declaration of Competing Interest The article was prepared and reviewed with the participation of all the authors. The authors declare no conflict of interest concerning this article's research, authorship, and publication., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Rickettsia felis is an emerging flea-borne spotted fever pathogen that causes febrile illness in humans. In Vietnam, R. felis was detected in hospitalized patients, but there is no information on its presence in the Vietnamese community. This cross-sectional study aimed to determine the presence of R. felis in humans of the Central Highlands of Vietnam. A total of 158 blood and 213 serum samples were subjected to PCR and IFAT, respectively, to detect the presence of R. felis DNA and antibodies against R. felis. PCR assays detected R. felis DNA in four out of 158 blood samples, accounting for a prevalence of 2.53 % (95 % CI: 0.81 %-6.76 %). Phylogenetic analysis indicated the presence of R. felis and R. felis genotype RF2125 in the communities in the Central Highlands of Vietnam. The result of IFAT identified seven out of 213 serum samples (3.29 %, 95 % CI: 1.45 %-6.93 %) positive for antibodies against R. felis. This study was the first to demonstrate the presence of active R. felis infections in the communities in the Central Highlands of Vietnam utilizing both molecular and serological methods., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Rickettsia felis, a zoonotic vector-borne bacteria, is reported globally in humans, animals, and its invertebrate hosts. This study was designed to detect antibodies against R. felis and the DNA of R. felis in blood of domestic dogs in the Central Highlands of Vietnam using immunofluorescence antibody test (IFAT), and ompB- and gltA-PCRs, respectively. Using IFAT, 23 out of 338 plasma samples collected from household dogs were seropositive for R. felis, accounting for 6.80% (CI 95%: 4.45-10.1%). Of 171 buffy coat samples from household dogs, 50 were positive for spotted fever group rickettsioses using ompB-PCR assay, accounting for 29.2% (CI 95%: 22.6-36.7%). The gltA-PCR assay detected R. felis in 30% (15/50) of ompB-positive samples. DNA sequencing of ompB-PCR and gltA-PCR products confirmed the presence of R. felis and Rickettsia sp. genotype RF2125 / R. asembonensis. Our findings suggest a potential risk of R. felis infection in the communities in the Central Highlands of Vietnam, and the reservoir role of dogs to Rickettsia sp. genotype RF2125., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

We report Rickettsia species from 2,334 ticks collected from environment (1,939 ticks) and animals (395 ticks) in the largest inland fragment of the Atlantic rainforest of southern Brazil and its fragments. Additionally, the DNA infection rates of Amblyomma ovale tick populations in the Neotropics with Rickettsia bellii and Rickettsia parkeri strain Atlantic rainforest were calculated using data from scientific publications, and their correlation was evaluated. From 11 tick species Rickettsia DNA was detected in seven (Amblyomma brasiliense, Amblyomma coelebs, Amblyomma incisum, Amblyomma longirostre, A. ovale, Haemaphysalis juxtakochi, Ixodes fuscipes) and was not detected in four species (Amblyomma dubitatum, Ixodes loricatus, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato). DNA of five Rickettsia species was detected (R. bellii, Rickettsia amblyommatis, Rickettsia rhipicephali, Rickettsia felis and Rickettsia sp. Aragaoi). To determine the prevalence of Rickettsia DNA positivity according to vector species, ticks were processed individually or in pools of 2-10 individuals (samples). The most prevalent Rickettsia species was R. bellii, found in 112 samples, followed by R. amblyommatis, R. rhipicephali, R. felis and Rickettsia sp. Aragaoi, found in 16, five, two and one sample, respectively. Rickettsia bellii DNA was found in five tick species with the highest infection rate in A. ovale and A. brasiliense. Absence of R. parkeri strain Atlantic rainforest in A. ovale ticks was an unexpected result. Furthermore, a negative correlation was identified between the infection rates (DNA) of R. bellii and/or R. parkeri strain Atlantic rainforest within A. ovale tick populations in the Neotropics. Putting together current knowledge, it can be proposed that, within natural settings, the diversity of rickettsiae and ticks creates a buffering effect on the overgrowth of rickettsiae and episodes of bacteremia in the hosts., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Mosquitoes are generally recognized as the most important vector of many zoonotic pathogens. In this study, seven mosquitoes species were identified (Anopheles pullus, Anopheles sinensis, Anopheles lesteri, Anopheles kleini, Ochlerotatus dorsalis, Aedes koreicus and Culex inatomii) in samples collected from Yingkou City, Liaoning Province, Northeastern China. A novel Rickettsia species was detected in Anopheles sinensis (two of 71, 2.82%) and Anopheles pullus (one of 106, 0.94%) mosquitoes. Genetic analysis indicated that the rrs and ompB genes have highest 99.60% and 97.88%-98.14% identities to Rickettsia felis, an emerging human pathogen of global concern mainly harboured by fleas, mosquitoes and booklice. The gltA sequences of these strains have 99.72% of nucleotide similarity with Rickettsia endosymbiont of Medetera jacula. The groEL sequences have 98.37% similarity to both Rickettsia tillamookensis and Rickettsia australis. The htrA sequences have 98.77% similarity to Rickettsia lusitaniae. In the phylogenetic tree based on concatenated nucleotide sequences of rrs, gltA, groEL, ompB and htrA genes, these strains are closely related to R. felis. Herein, we name it 'Candidatus Rickettsia yingkouensis'. Its human pathogenicity to humans and animals is still to be determined., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

The Eastern Uttar Pradesh region of India is known for its endemicity of acute encephalitis syndrome (AES). Decades of research have established that Orientia tsutsugamushi, a causative of scrub typhus, is a substantial contributor (>60%) for the AES cases besides other aetiology, but additional factors in the remaining proportion are still unidentified. Rickettsial infections are challenging to diagnose in clinical settings due to overlapping clinical symptoms, the absence of definitive indicators, a low index of suspicion, and the lack of low-cost, rapid diagnostic tools. Hence, the present study was designed to determine the load of rickettsial infections among AES cases. Furthermore, we aim to find out the prevalent rickettsial species in AES cases as well as in the vector population at this location. The study included the whole blood/cerebrospinal fluid of AES patients and arthropod specimens from rodents. The molecular identification was performed using the 23S-5S intergenic spacer region and ompB gene with genomic DNA obtained from studied specimens. We detected 5.34% (62/1160) of rickettsial infection in AES cases. Among these, phylogenetic analysis confirmed the presence of 54.8% Rickettsia conorii (n = 34) and 16.1% of Rickettsia felis (n = 10), while the rest proportion of the isolates was unidentified at the species level. Furthermore, R. felis was identified in one CSF sample from AES patients and three flea samples from Xenopsylla cheopis. Rickettsia spp. was also confirmed in one Ornithonyssus bacoti mite sample. The results of this investigation concluded the presence of spotted fever group Rickettsia spp. among AES identified cases as well as in the mite and flea vectors that infest rodents., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Tick-borne pathogens (TBPs) pose a significant threat to livestock, including bovine species. This study aimed to investigate TBPs in cattle and ticks across four sampling points, utilizing real-time microfluidic PCR. The results revealed that Rhipicephalus microplus ticks were found infesting all animals. Among the detected TBPs in cattle, Anaplasma marginale was the most frequently identified, often as a single infection, although mixed infections involving Rickettsia felis , uncharacterized Rickettsia sp., and Anaplasma sp. were also observed. In ticks, A. marginale was predominant, along with R. felis , Rickettsia sp., and Ehrlichia sp. It is noteworthy that although A. marginale consistently infected all cattle during various sampling times, this pathogen was not detected in all ticks. This suggests a complex dynamic of pathogen acquisition by ticks. A phylogenetic analysis focused on the identification of Anaplasma species using amplified 16S rDNA gene fragments revealed the presence of A. marginale and Anaplasma platys strains in bovines. These findings underscore the presence of multiple TBPs in both cattle and ticks, with A. marginale being the most prevalent. Understanding the dynamics and phylogenetics of TBPs is crucial for developing effective control strategies to mitigate tick-borne diseases in livestock.

Fleas are important ectoparasites and vectors associated with a wide range of pathogenic diseases, posing threats to public health concerns, especially cat fleas that spread worldwide. Understanding the microbial components is essential due to cat fleas are capable of transmitting pathogens to humans, causing diseases like plague and murine typhus. In the present study, metagenomic next-generation sequencing was applied to obtain the complete microbiota and related functions in the gut of Ctenocephalides felis. A total of 1,870 species was taxonomically recognized including 1,407 bacteria, 365 eukaryotes, 69 viruses, and 29 archaea. Proteobacteria was the dominant phylum among the six samples. Pathogens Rickettsia felis, Acinetobacter baumannii, Coxiella burnetii, and Anaplasma phagocytophilum were taxonomically identified and had high abundances in all samples. The resistance gene MexD was predominant in microbial communities of all cat fleas. We also performed epidemiological surveys of pathogens R. felis, A. baumannii, C. burnetii, and A. phagocytophilum among 165 cat fleas collected from seven provinces in China, while only the DNAs of R. felis (38/165, 23.03%) and C. burnetii (2/165, 1.21%) were obtained. The data provide new insight and understanding of flea intestinal microbiota and support novel information for preventing and controlling fleas and their transmitted diseases., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Rickettsia is an important pathogenic entity among tick-borne diseases (TBD), which are considered serious emerging public health problems globally. In India, though the widespread distribution of ticks and TBD has been documented, its real burden remains underreported. In a preliminary attempt, rickettsial surveillance was carried out in ticks collected from Sirumalai, Eastern Ghats in Tamil Nadu, India by using pathogen genome-based phylogenetic inferences generated through multi-locus sequence typing (MLST), targeting the genes 16s rRNA, OmpA, OmpB, and gltA by nested PCR. The laboratory evidence confirms the circulation of Rickettsia in Haemaphysalis intermedia species collected from this area. Analysis of the four gene sequences detected demonstrates their closest identity to the spotted fever group (SFG) available in the GenBank database. Further, multiple sequence alignment with other sequences derived from the GenBank database showed close relatedness to Rickettsia conorii subsp. raoultii (16s rDNA-99.32%, OmpA-93.38%, OmpB-97.39%, and gltA-98.57%) and Rickettsia felis (16s rDNA 99.54%, OmpA-100%, OmpB-100% and gltA-99.41%). With this genomic evidence, the circulation of rickettsial pathogens in the pools of H. intermedia ticks infesting livestock in the Sirumalai foothill area has been demonstrated and to complement the microscopic identification of the tick species, DNA barcodes were generated for H. intermedia using the mitochondrial cytochrome c oxidase subunit I gene (COI). Nevertheless, R. raoultii and R. felis were found to be the aetiological agents of tick-borne lymphadenopathy and flea-borne spotted fever in human cases, respectively, further study on the determination of their diversity, distribution, clinical relevance, and potential risk to the local community in these areas is highly warranted.

Neglected bacterial zoonoses are a group of Neglected Tropical Diseases (NTDs) that are commonly underdiagnosed and underreported due to their undifferentiated febrile illness symptomology. Spotted fever group rickettsioses (SFGR), a subset of tick-borne bacterial zoonoses, belong in this group. There is a dichotomy in the reporting and recognition of these pathogens in Central America: countries with reduced human development scores-like El Salvador-have little to no research or surveillance dedicated to these pathogens and the diseases they cause. This was the third-ever tick survey in El Salvador, highlighting the knowledge gap in this country. A total of 253 ticks were collected from 11 animals at two farm sites and one veterinary office. Standard and quantitative PCR were used to detect presence of SFGR, Ehrlichia, and Anaplasma sp. pathogens in ticks. Ehrlichia sp. were detected in 2.4% of all collected ticks and Anaplasma sp. were detected in 5.5% of all ticks. Rickettsia rickettsii was amplified in 18.2% of ticks, and amplicons similar to R. parkeri, and R. felis were found in 0.8% and 0.4%, of collected ticks, respectively. This is the first report of these pathogenic bacterial species in El Salvador. This study emphasizes the need for further surveillance and research including incorporating additional human seroprevalence and testing to understand the public health burden in this country., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Seventy-five flea pools (one to ten fleas per pool) from 51 Andean foxes (Lycalopex culpaeus) and five South American grey foxes or chillas (Lycalopex griseus) from the Mediterranean region of Chile were analyzed for the presence of DNA of Bartonella spp. and Rickettsia spp. through quantitative real-time PCR for the nouG and gltA genes, respectively. Positive samples were further characterized by conventional PCR protocols, targeting gltA and ITS genes for Bartonella, and gltA, ompA, and ompB genes for Rickettsia. Bartonella was detected in 48 % of the Pulex irritans pools (B. rochalimae in three pools, B. berkhoffii in two pools, B. henselae in one pool), and 8 % of the Ctenocephalides felis felis pools (B. rochalimae, one pool). Rickettsia was confirmed in 11 % of P. irritans pools and 92 % of the Ct. felis pools. Characterization confirmed R. felis in all sequenced Rickettsia-positive pools. All Ct. canis pools were negative. A Ct. felis pool from a wild-found domestic ferret (Mustela putorius furo) also resulted positive for R. felis. Although opportunistic, this survey provides the first description of zoonotic pathogens naturally circulating in fleas parasitizing Chilean free-living carnivores., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Surveillance of the fleas and flea-borne pathogens infecting cats is important for both human and animal health. Multiple zoonotic Bartonella and Rickettsia species are known to infect the most common flea infesting cats and dogs worldwide: Ctenocephalides felis , the cat flea. The ability of other flea species to transmit pathogens is relatively unexplored. We aimed to determine cat host and flea factors independently associated with flea Bartonella and Rickettsia infection. We also assessed flea and cat infection by flea-host pair and location. To accomplish these aims, we performed qPCR for the detection of Bartonella , hemotropic Mycoplasma , Rickettsia , and Wolbachia DNA using paired cat and flea samples obtained from free-roaming cats presenting for spay or neuter across four locations in the United States. A logistic regression model was employed to identify the effect of cat (sex, body weight, geographic location, and Bartonella , hemotropic Mycoplasma , and Rickettsia spp., infection) and flea (clade and Rickettsia and Wolbachia infection) factors on C . felis Bartonella clarridgeiae infection. From 189 free roaming cats, we collected 84 fleas: Ctenocephalides felis (78/84), Cediopsylla simplex (4/84), Orchopeas howardi (1/84), and Nosopsyllus fasciatus (1/84). Ctenocephalides felis were phylogenetically assigned to Clades 1, 4, and 6 by cox1 gene amplification. Rickettsia asembonensis (52/84) and B . clarridgeiae (16/84) were the most common pathogenic bacteria detected in fleas. Our model identified host cat sex and weight as independently associated with B . clarridgeiae infection in fleas. Rickettsia asembonensis (52/84), Rickettsia felis (7/84) and Bartonella henselae (7/84) were detected in specific clades: R . felis was detected only in Clades 1 and 6 while B . henselae and R . asembonensis were detected only in Clade 4. Wolbachia spp., also displayed clade specificity with strains other than Wolbachia wCfeT only infecting fleas from Clade 6. There was poor flea and host agreement for Bartonella spp., infection; however, there was agreement in the Bartonella species detected in cats and fleas by geographic location. These findings reinforce the importance of considering reservoir host attributes and vector phylogenetic diversity in epidemiological studies of flea-borne pathogens. Widespread sampling is necessary to identify the factors driving flea-borne pathogen presence and transmission., Competing Interests: In conjunction with S. Sontakke and North Carolina State University, EB holds US Patent No. 7,115,384 Media and Methods for Cultivation of Microorganisms, which was issued on October 3rd, 2006, and also co-founder, shareholder, and Chief Scientific Officer for Galaxy Diagnostics, a company that provides advanced diagnostic testing for the detection of Bartonella spp. infections. The handling editor MA declared a past co-authorship with the EB and RM. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Moore, Breitschwerdt, Kim, Li, Ferris, Maggi and Lashnits.)

The Stone Age record of South Africa provides some of the earliest evidence for the biological and cultural origins of Homo sapiens. While there is extensive genomic evidence for the selection of polymorphisms in response to pathogen-pressure in sub-Saharan Africa, e.g., the sickle cell trait which provides protection against malaria, there is inadequate direct human genomic evidence for ancient human-pathogen infection in the region. Here, we analysed shotgun metagenome libraries derived from the sequencing of a Later Stone Age hunter-gatherer child who lived near Ballito Bay, South Africa, c. 2000 years ago. This resulted in the identification of ancient DNA sequence reads homologous to Rickettsia felis, the causative agent of typhus-like flea-borne rickettsioses, and the reconstruction of an ancient R. felis genome., (© 2023. The Author(s).)

Fleas and ticks represent the two main groups of ectoparasites that infest companion animals. In particular, the flea Ctenocephalides felis felis and several members of the Rhipicephalus sanguineus complex are the main vectors of a wide range of pathogens on the American continent. They are competent vectors for several members of the genus Rickettsia, which encompass at least 15 pathogenic obligate intracellular bacteria that colonize the endothelial cells of vertebrates. In Mexico, 10 species of Rickettsia belonging to three groups have been detected in six species of ectoparasites from dogs in 9 of the 32 states of the country. However, in some larger regions of the country, active epidemiological surveillance has not been carried out. For this reason, the aim of this study was to identify the presence of members of the genus Rickettsia in fleas and ticks of dogs and cats in the state of Puebla, Mexico. A cross-sectional study was carried out to collect ectoparasites of dogs and cats during August to November 2019. Samples were fixed in 70% ethanol and examined to identify the presence of Rickettsia DNA by the amplification and sequencing of specific fragments of the gltA and ompB genes using conventional PCR. The recovered sequences were compared with those deposited in GenBank, and phylogenetic analyses were carried out to identify the position of the pathogens detected with respect to the valid species previously reported worldwide. Additionally, ecological parameters of the ectoparasite infestations were also calculated. We recovered 196 ectoparasites belonging to two species, 33 C. felis felis and 163 R. sanguineus s.l. (Rhipicephalus linnaei), parasitizing 46 hosts (42 dogs and 4 cats) in 11 localities of the state of Puebla. We detected the presence of Rickettsia felis in three pools of C. felis felis, and five from R. sanguineus s.l. Our work provides the first record of R. felis in hard ticks of Mexico and Central America, with new collection localities for this pathogen in central Mexico., (© 2022 Wiley-VCH GmbH.)

Rickettsia felis is an emerging rickettsial agent principally associated with cat fleas (Ctenocephalides felis), formerly discovered in 1990. Since then, clinical cases of R. felis infection have been identified globally by specific DNA sequences in patients with undifferentiated febrile illness, including in Taiwan, but such evidence is limited. R. felis rickettsiosis is self-limiting and easily treated with doxycycline, but its diagnosis remains a challenge. Environmental risk factors for R. felis rickettsiosis have yet to be clearly demonstrated, and its transmission biology is incompletely understood. Cat fleas are naturally infected with R. felis at varying rates, and vector competence in the transmission of R. felis has been demonstrated in animal models, including dogs, which may serve as reservoir hosts. In northern Taiwan, despite ∼20% of cat fleas infesting companion animals consistently found to be infected with R. felis, only a few cases of potential R. felis infection have been identified through a retrospective serological investigation, though without molecular confirmation. Ecological studies have identified divergent R. felis-like organisms in different arthropod hosts, but these strains appear to serve as nonpathogenic endosymbionts. Although its association with disease is limited, we believe cat flea-borne R. felis warrants increased recognition in an aging population due to immunosenescence and the proximity of companion animals to the elderly. Adopting a One Health approach involving collaboration and communication between clinicians, veterinarians, public health practitioners, and environmental scientists will improve our knowledge about this neglected pathogen and promote the prevention and control of vector-borne diseases., (Copyright © 2022. Published by Elsevier B.V.)

Background: In recent years, promising vaccination strategies against rickettsiosis have been described in experimental animal models and human cells. OmpB is considered an immunodominant antigen that is recognized by T and B cells. The aim of this study was to identify TCD4+INF-γ+ and TCD8+INF-γ+ lymphocytes in an autologous system with macrophages transfected with the vaccine candidate pVAX1-OmpB24. Lymphocytes and monocytes from 14 patients with Rickettsia were isolated from whole blood. Monocytes were differentiated into macrophages and transfected with the plasmid pVAX1-OmpB24 pVax1. Isolated lymphocytes were cultured with transfected macrophages. IFN-γ-producing TCD4+ and TCD8+ lymphocyte subpopulations were identified by flow cytometry, as was the percentage of macrophages expressing CD40+, CD80+, HLA-I and HLA-II. Also, we analyzed the exhausted condition of the T lymphocyte subpopulation by PD1 expression. Macrophages transfected with pVAX1-OmpB24 stimulated TCD4+INF-γ+ cells in healthy subjects and patients infected with R. typhi . Macrophages stimulated TCD8+INF-γ+ cells in healthy subjects and patients infected with R. rickettsii and R. felis . Cells from healthy donors stimulated with OmpB-24 showed a higher percentage of TCD4+PD1+. Cells from patients infected with R. rickettsii had a higher percentage of TCD8+PD-1+, and for those infected with R. typhi the larger number of cells corresponded to TCD4+PD1+. Human macrophages transfected with pVAX1-OmpB24 activated TCD4+IFN-γ+ and CD8+IFN-γ+ in patients infected with different Rickettsia species. However, PD1 expression played an important role in the inhibition of T lymphocytes with R. felis.

Chiggers are larval ectoparasites of the Trombiculidae that can transmit pathogens to their hosts. In this study, chiggers collected from birds in Brazil were morphologically identified as Blankaartia sinnamaryi, Eutrombicula batatas, Eutrombicula daemoni, Eutrombicula goeldii, Eutrombicula tinami, and Parasecia gilbertoi. For these specimens, a beginning attempt at molecular identification were also provided, as well as, were genetically screened to detect bacterial pathogens. The species B. sinnamaryi and E. tinami were positive for Rickettsia felis-like and 'Candidatus Rickettsia colombianensi'-like, respectively. For the other agents (Anaplasmataceae, Borrelia spp. and Orientia tsutsugamushi), the tests were negative. This is the first report of 'Ca. R. colombianensi'-like and the second record of R. felis-like in chigger collected on birds from Brazil., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Rickettsia felis is an obligate intracellular gra m-negative bacterium that belongs to the family of Rickettsiaceae. Ctenocephalides felis, cat flea, is the primary vector of the bacteria. The flea is the most common ectoparasite in dogs and associated with flea-borne spotted fever in humans. Information on R. felis and flea species parasitizing on dogs in Vietnam is limited. This study aimed to identify the species of fleas collected from dogs in the Central Highlands of Vietnam and detected the existence of R. felis in these fleas utilizing molecular tools. Morphological identification of 1618 fleas and molecular confirmation revealed the predominance of C. felis orientis parasitizing on dogs in the Central Highlands of Vietnam. Sixty-eight out of 100 fleas collected from household dogs were positive for spotted fever group rickettsiae; whilst R. felis was detected in 97.06 % (66/68) of C. felis orientis and C. felis felis. The results of this study indicate the potentially high risk of R. felis infection to humans and animals., Competing Interests: Conflict of interest The authors declare no conflicts of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)