Rickettsial Pathogens Research Articles

Phosphatidylserine-binding receptor, CD300f, on macrophages mediates host invasion of pathogenic and non-pathogenic rickettsiae.

Some arthropod-borne obligate intracellular rickettsiae are among the most virulent human pathogens. Rickettsia species modulate immune (e.g., macrophages; MΦ) and non-immune cell (e.g., endothelial cells) responses to create a habitable environment for host colonization. In particular, MΦ play a crucial role in either terminating an infection at an early stage or succumbing to bacterial replication and colonization. However, our understanding on how Rickettsia species invade host cells, including MΦ, remain poorly defined. In this study, we describe a mechanism of host invasion by Rickettsia species, involving rickettsial phosphatidylserine (PS), as a ligand, and the CD300f receptor on MΦ. Using bone marrow-derived macrophages (BMDMΦ) from wild-type (WT) and CD300f-/- mice, we demonstrated that engulfment of both pathogenic R. typhi (the etiologic agent of murine typhus) and R. rickettsii (the etiologic agent of Rocky Mountain spotted fever) species as well as the non-pathogenic R. montanensis was significantly reduced in CD300f-/- BMDMΦ as compared to that of WT BMDMΦ. Furthermore, our mechanistic analysis suggests bacterial PS as the potential source for the CD300f-mediated rickettsiae engulfment by MΦ. In vivo infection studies using WT and CD300f-/- C57BL/6J mice showed that CD300f-/- animals were protected against R. typhi- or R. rickettsii-induced fatal rickettsiosis, which correlated with levels of bacterial burden detected in the spleens of mice. Adoptive transfer studies further revealed that CD300f-expressing MΦ are important mediators to control rickettsiosis in vivo. Collectively, our findings describe a previously unappreciated role for the efferocytic receptor, CD300f, to facilitate engulfment of rickettsiae within the host.

Development of a Recombinase-Mediated Cassette Exchange System for Gene Knockout and Expression of Non-Native Gene Sequences in Rickettsia

Background/Objectives: Incidence of vector-borne diseases, including rickettsioses and anaplasmosis, has been increasing in many parts of the world. The obligate intracellular nature of rickettsial pathogens has hindered the development of robust genetic tools for the study of gene function and the identification of therapeutic targets. Transposon mutagenesis has contributed to recent progress in the identification of virulence factors in this important group of pathogens. Methods: Combining the efficiency of the himar1 transposon method with a recombinase-mediated system, we aimed to develop a genetic tool enabling the exchange of the transposon with a cassette encoding non-native sequences. Results: This approach was used in Rickettsia parkeri to insert a himar1 transposon encoding fluorescent protein and antibiotic resistance genes for visualization and selection, flanked by mismatched loxP sites to enable subsequent recombinase-mediated cassette exchange (RMCE). RMCE mediated by a plasmid-encoded Cre recombinase was then employed to replace the transposon with a different cassette containing alternate fluorescent and selection markers and epitopes of Anaplasma phagocytophilum antigens. The resulting genetically modified R. parkeri was trialed as a live-attenuated vaccine against spotted fever rickettsiosis and anaplasmosis in mice. Conclusions: The use of this system provides a well-established and relatively efficient way of inserting non-native sequences into the rickettsial genome, with applications for the study of gene function and vaccine development.

Rickettsia parkeri forms extensive, stable contacts with the rough endoplasmic reticulum.

Upon invasion into the host cell, a subset of bacterial pathogens resides exclusively in the cytosol. While previous research revealed how they reshape the plasma membrane during invasion, subvert the immune response, and hijack cytoskeletal dynamics to promote their motility, it was unclear if these pathogens also interacted with the organelles in this crowded intracellular space. Here, we examined if the obligate intracellular pathogen Rickettsia parkeri interacts with the endoplasmic reticulum (ER), a large and dynamic organelle spread throughout the cell. Using live-cell microscopy and transmission and focused-ion-beam scanning electron microscopy, we show that R. parkeri forms extensive contacts with the rough ER that are ∼55 nm apart and cover more than half the bacterial surface. Depletion of the ER-specific tethers VAPA and VAPB reduced rickettsia-ER contacts, and VAPA and VAPB were localized around intracellular rickettsiae. Overall, our findings illuminate an interkingdom ER contact uniquely mediated by rickettsiae that mimics some characteristics of traditional host membrane contact sites.

First report of Anaplasma marginale and Anaplasma ovis in goats in Kelantan, Malaysia.

Anaplasma species are obligate rickettsial intraerythrocytic pathogens that cause an important tick-borne disease of economic importance in livestock production in many countries. Anaplasma species have been detected from farm animals worldwide, there is a paucity of information on Anaplasma infections in goats from Malaysia. Thus, this study aimed to assess the infection rate and identify Anaplasma species and some selected risk factors in goats across selected districts in Kelantan, Malaysia. A total of 411 blood samples were collected from goats and analysed for Anaplasma species targeting the msp4 gene using conventional PCR and sequencing. The infection risk was determined by breed, age, management system and location. Our results indicate an overall infection rate of 30.9% for Anaplasma species detected. Interestingly, sequencing of selected amplicons revealed the presence of Anaplasma ovis and A. marginale. Data analysis revealed a marked statistically significant association between Anaplasma infection and some variables such as location (district), farm management system, breed, and age (P < 0.05). Specifically, goats raised on intensive management had the highest prevalence of 46.25% (37/80) compared to other management types. Also, with regards to district, goats raised in the coastal region had a higher prevalence of 39.23% (71/181) compared to those raised in inland region 24.35% (56/230). Regarding breed, goats that were of the pure breed had a higher prevalence of Anaplasma species infection 38.19% (97/254) compared to crossbreeds with a prevalence of 19.11% (30/157). Lastly, goats 3 years had the least prevalence 18.99% (34/179). To the best of our knowledge, this is the first report of A. marginale and A. ovis in goats from northeastern Peninsular Malaysia. The infected goats were clinically healthy; this revealed the role of goats as a potential reservoir for A. marginale and the presence of A. ovis in goats in Malaysia. Continuous efforts towards tick control must be sustained to ensure high productive yield and reduced disease burden associated with TBPs of goats in the study area.

Molecular Detection of Tick-Borne Bacterial Pathogens in Patients With Undifferentiated Febrile Illness in India Using a Flow Chip Assay.

Tick-borne diseases (TBDs) play a crucial role in human morbidity and mortality, as ticks are highly effective in spreading diseases by transmitting harmful pathogens to humans and animals. The last few decades have seen an increase in the number of recognized tick-borne pathogens and the incidence of TBD worldwide. Several of these diseases are ubiquitous in India. However, in India, there is limited information on the molecular detection of tick-borne pathogens in patients with undifferentiated febrile illness. The study aims to investigate tick-borne pathogens among undifferentiated febrile patients in India using a multiplex polymerase chain reaction (PCR)-based assay. A total of 120 blood samples were collected from patients with undifferentiated febrile illnesses of all ages. The samples were tested for a panel of (seven) tick-borne pathogens (Anaplasma, Ehrlichia, Borrelia, Bartonella, Coxiella, Rickettsia, and Francisella) using a multiplex PCR tick-borne bacterial flow chip (TBFC) assay through a semi-automated HybriSpot platform (Vitro Master Diagnóstica, Granada, Spain). Among the 120 samples tested, one was positive for spotted feverRickettsia, two were positive for typhus groupRickettsia, one was positive forBorrelia, and two showed coinfections withBorreliaandAnaplasma. Our knowledge of TBD is steadily advancing with the discovery of novel pathogens and the development of cutting-edge diagnostic tools. Although traditional diagnostic methods like serology and microscopy will remain essential for the diagnosis of TBD, the implementation of advanced molecular diagnostics such as TBFC will enhance our understanding of these diseases by enabling the identification of emerging pathogens and offering more precise and timely diagnoses.

Molecular detection of Anaplasma platys in Pshdar Kurdish shepherd dogs of Sulaymaniyah Province, Iraq.

Anaplasma platys is a dog pathogen that causes anaplasmosis in various hosts, including humans. It is a rickettsial pathogen that causes cyclic thrombocytopenia in primary canine recipients and is spread by the brown dog tick Rhipicephalus sanguineus. This study aimed to map the genetic sequences of Anaplasma spp. isolates comparable with those of different global locations and determine the infection status of Pshdar Kurdish shepherd dogs from three regions in Sulaymaniyah province who did not exhibit clinical indications for Anaplasma. A total of 75 dog blood samples were collected from the center of the Sulaymaniyah, Dukan, and Ranya districts in the Sulaymaniyah province and subjected to polymerase chain reaction to determine the 16S ribosomal RNA (16S rRNA) gene of A. platys. Only two dogs (2.7%) were positive for A. platys. The 16S rRNA gene of A. platys was sequenced and registered in the National Center for Biotechnology Information GenBank with accession number OR467538. With four nucleotide changes, the sequence exhibited 99.72% similarity to strains identified as human infections and those found in recognized tick vectors. We conclude that the blood of Pshdar Kurdish shepherd dogs in the Sulaymaniyah region of Iraq contains a small number of A. platys. Moreover, the phylogenetic tree of the isolated species, A. platys, was significantly similar to other strains of A. platys found worldwide. In the Kurdistan region of Iraq, this study represents the first molecular detection of the 16S rRNA gene of A. platys.

Tick-borne pathogens in Mongolian ticks: The high prevalence of Rickettsia raoultii and its public health implications.

In Mongolia, a substantial segment of the population is engaged in pastoralism, leading to frequent interactions with livestock and heightened exposure to tick-infested environments. Consequently, ticks and the endemic tick-borne pathogens (TBPs) they harbor present significant health threats. In May 2021, we conducted a study to evaluate the prevalence of 9 TBPs in ticks collected from four regions in Mongolia: Uvurkhangai, Tuv, Bayan-Ulgii, and Khentii. The pathogens surveyed included Anaplasma, Rickettsia, Bartonella, Borrelia, Ehrlichia, Babesia, Toxoplasma, Theileria, and lumpy skin disease virus. Molecular analysis of 1142 ticks revealed that, although the majority of TBPs were not detected, 89.1 % of the ticks were positive for Rickettsia. Genetic characterization using the 16S rRNA and gltA genes identified the pathogen identified the pathogen Rickettsia raoultii. Species identification indicated a predominance of Dermacentor nuttalli (70.0 %) and Dermacentor silvarum (30.0 %). These findings highlight the extensive prevalence of R. raoultii in Mongolia and underscore the urgent need for heightened awareness and preventive measures due to the high level of interaction between livestock and humans. The genetic similarity of R. raoultii sequences to those found in neighboring countries suggests potential cross-border transmission, underscoring the importance of conducting similar research in surrounding regions. This study advances our understanding of the epidemiology of tick-borne diseases in Mongolia and can inform public health strategies aimed at mitigating the risks associated with these infections.

Prevalence and diversity of spotted fever group Rickettsia species in ixodid ticks from domestic dogs in Chad, Africa

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.

Microbial community characteristics and pathogens detection in Rhipicephalus sanguineus and Haemaphysalis hystricis from Hainan Island, China.

Microbial communities significantly influence the vector capacity of ticks, which, along with tick-borne diseases, pose an increasing global threat. Due to the substantial individual variability caused by various factors, it is essential to assess tick microbial communities and vectorial capacities under different environmental conditions. However, there is a relative scarcity of research on the microbial communities and pathogen transmission of ticks in different physiological states and environmental conditions, especially in Hainan Island, southern China. From 2021 to 2022, we collected 4,167 tick samples, grouping them by blood meal status, developmental stage, sex, time, geographical location, and tick species. We selected 128 samples for full-length 16S rRNA sequencing to describe microbial community characteristics and identify potential biomarkers. Seven hundred seventy-two samples were tested for seven tick-borne pathogens (Rickettsia, Borrelia burgdorferi, Ehrlichia, Anaplasma, Theileria, Babesia, and Hepatozoon), and sera from 208 residents of Hainan Island were tested for IgG antibodies against Rickettsia and B. burgdorferi. Blood meal status, developmental stage, sex, time, geographical location, and tick species significantly influenced the microbial communities of ticks. We observed distinct microbial community characteristics across different states. We noted the non-random replacement of stable and transient species, with functional differences between parasitic and engorged ticks mainly driven by transient species. Functionally, we observed three distinct response patterns: driven by stable species, transient species, and both together in response to the six factors. We identified 273 potential biomarkers (200 robust core species and 73 robust differential species). Six genera and eight species of pathogens were detected in ticks, with an overall positivity rate of 12.44% (96/772). Among humans, 18.27% (38/208) of serum samples were positive for at least one tick-borne pathogen IgG. Our findings indicate that these six factors significantly influence both tick microbial communities and vectorial capacity, with varying effects on vector competence for different pathogens and inconsistent impacts on microbial communities under different conditions. This study supplemented the understanding of tick microbial communities on Hainan Island, assessed the relatively high risk of tick-borne pathogens in the region, and evaluated the impact of these factors on both microbial communities and vectorial capacity.

First molecular diagnosis of the human pathogen Rickettsia raoultii and other spotted fever group rickettsiae in Sudanese ixodid ticks from domestic ruminants.

Rickettsial infections are often neglected and poorly recognized by physicians in many tropical and subtropical regions. Despite a number of recent reports describing rickettsial diseases in new locations and the discovery of new rickettsiae, medical science and research have largely neglected the diagnosis and antimicrobial treatment of rickettsial infections in subtropical and tropical areas; thus, much remains to be discovered. This study aimed to detect and characterize spotted fever group (SFG) rickettsiae in ixodid ticks infesting domestic ruminants in Khartoum State. Polymerase chain reaction targeting both genes that encode for citrate synthase (gltA) and outer membrane protein (ompA) was performed for the presence of SFG rickettsia followed by sequence and phylogenetic analysis. Of the 202 ticks examined for the presence of SFG rickettsia, gltA gene was detected in 4 samples (2%). Furthermore, gltA-positive samples were used to amplify the ompA gene, in which only two samples yielded positive results. Sequence and phylogenetic analysis of the positive samples revealed four different species of SFG rickettsiae: Rickettsia aeschlimannii, Rickettsia rhipicephali, Rickettsia massiliae and Rickettsia raoultii. These results indicated the presence of SFG rickettsia in Sudanese ticks. This also indicates that humans have an opportunity to acquire these infections. It is important to keep in mind the need for careful consideration of rickettsial infections in individuals with a fever of unknown origin.

Habitat Suitability of Ixodes ricinus Ticks Carrying Pathogens in North-East Italy.

Ixodes ricinus ticks are ubiquitous in Europe, including in North-East Italy. These ticks are important vectors of several zoonotic pathogens of public health relevance. In this study, the habitat suitability range of I. ricinus ticks infected with zoonotic pathogens was predicted in North-East Italy, and relevant spatial predictors were identified. In 2015-2021, ticks were collected at 26 sampling sites in the study area. The collected ticks were screened for the presence of pathogens using PCR assays. For Borrelia, Rickettsia and Anaplasma/Ehrlichia species, data allowed for ecological niche modelling using Maxent. Environmental determinants potentially related to tick habitat suitability were used as model inputs. Predicted suitable habitat distributions revealed hotspots of the probability of pathogen presence in I. ricinus ticks mainly in the central and upper parts of the study area. Key environmental predictors were temperature, rainfall and altitude, and vegetation index for specific pathogens (Rickettsia and Anaplasma/Ehrlichia species). Increased risk of exposure to tick-borne pathogens upon tick bites in the predicted hotspot areas can, therefore, be expected. This provides useful information for public health risk managers in this and other similar regions.

WITHDRAWN: Detection of Amblyomma maculatum and Rickettsia parkeri in prairies of Central Missouri

Prairie habitats are complex ecosystems experiencing destruction and decline due to landscape repurposing, thus resulting in significant changes to plant, animal, and insect wildlife diversity. In the Central United States, the reconstruction of prairie habitats from farmland is a widely applied strategy to raise diversity and recreate a healthy, complex ecosystem. In Central Missouri, we are examining the consequences of reconstruction efforts on the prevalence of zoonotic diseases and their associated pathogens and have observed large populations of Dog ticks, Dermacentor variabilis, and Lone Star ticks, Amblyomma americanum, as well as the first recorded instance of the Gulf Coast tick, Amblyomma maculatum, in Callaway County. The Gulf Coast tick is traditionally found mainly along the Atlantic and Gulf Coast regions, but has been undergoing geographic expansion in the last decade. Since detection in 2020, the number of Gulf Coast ticks has increased,. Upon screening 193 Gulf Coast ticks in 105 pools, we found that 6.67 % of ticks were positive for the pathogenic Rickettsia parkeri, which causes spotted fever rickettsiosis in animals and humans, with most ticks coming from the reconstructed prairie site. These observations suggest that reconstruction of prairie habitat has the potential to support the sylvatic cycle of Rickettsia parkeri. Collectively, our observations show that the reconstructed prairie are capable of harboring large tick populations as compared to remnant prairies and demonstrates a potential increase in disease risk as a result. This works highlights the importance of proactive surveillance of ticks, especially with land-use change.

Dog ectoparasites as sentinels for pathogenic Rickettsia and Bartonella in rural Guatemala

Fleas and ticks serve as vectors of multiple pathogens in the genera Rickettsia and Bartonella that cause diseases in humans and other animals. Although human rickettsiosis and bartonellosis have been reported in all countries in Central America, limited research has been conducted to investigate the natural cycles of flea- and tick-borne rickettsiosis and bartonellosis, especially in Guatemala. We evaluated dog parasites as sentinels for zoonotic disease risk in rural Guatemala by sampling ticks and fleas from dogs, which were then identified and individually screened for Rickettsia and Bartonella. A total of 77 households were surveyed and 80.5 % of them had dogs. Overall, 133 dogs were examined for fleas and ticks, of which 68.4 % had fleas and 35.3 % had ticks. A total of 433 fleas and 181 ticks were collected from the infested dogs, with an additional 33 ticks collected from house walls. Three flea species were identified: Ctenocephalides felis (70.0 %), Echidnophaga gallinacea (11.8 %), and Pulex sp. (17.8 %). Among the collected ticks, 97 % were identified as Rhipicephalus sanguineus sensu lato with the rest being Amblyomma cajennense, A. auricularium, and A. ovale. Rickettsia felis were detected in six C. felis, in one Pulex sp., and in two R. sanguineus sensu lato, while Candidatus R. senegalensis was detected in one C. felis. Bartonella was detected only in fleas, including three Pulex sp. infected with B. vinsonii subsp. berkhoffii, B. henselae, and Bartonella sp., respectively, and 11 C. felis infected with B. henselae. This study reports Candidatus R. senegalensis and B. vinsonii subsp. berkhoffii in Guatemala for the first time, and indicates the potential risk of human and dog exposure to Rickettsia and Bartonella species. These results show that dogs provide critical information relevant to managing human potential exposure to flea- and tick-borne pathogens in rural Guatemala. This approach can potentially be expanded to other regions in Central America where domestic dogs are abundant and suffer from ectoparasite infestation.

Genomic characterization of an emerging Rickettsia barbariae isolated from tick eggs in northwestern China

ABSTRACT The continual emergence of tick-borne rickettsioses has garnered widespread global attention. Candidatus Rickettsia barbariae (Candidatus R. barbariae), which emerged in Italy in 2008, has been detected in humans from northwestern China. However, the lack of Candidatus R. barbariae genome and isolated strains limits the understanding of its biological characteristics and genomic features. Here, we isolated the Rickettsia for the first time from eggs of Rhipicephalus turanicus in northwestern China, and assembled its whole genome after next-generation sequencing, so we modified the proposed name to Rickettsia barbariae (R. barbariae) to conform to the International Code of Nomenclature of Prokaryotes. Phylogenetic analysis based on the whole genome revealed that it was most closely related to the pathogenic Rickettsia parkeri and Rickettsia africae. All virulence factors, present in the pathogenic spotted fever group rickettsiae, were identified in the R. barbariae isolate. These findings highlight the pathogenic potential of R. barbariae and the necessity for enhanced surveillance of the emerging Rickettsia in the human population.

Long-Term Protective Immunity against Ehrlichia chaffeensis Infection Induced by a Genetically Modified Live Vaccine.

Human monocytic ehrlichiosis, an emerging tick-borne disease, is caused by Ehrlichia chaffeensis. Infections with the pathogen are also common in the canine host. Our previous studies demonstrated that functional disruption within the E. chaffeensis phage head-to-tail connector protein gene results in bacterial attenuation, creating a modified live attenuated vaccine (MLAV). The MLAV confers protective immunity against intravenous and tick transmission challenges one month following vaccination. In this study, we evaluated the duration of MLAV protection. Dogs vaccinated with the MLAV were challenged with wild-type E. chaffeensis via intravenous infection at 4-, 8-, and 12-months post-vaccination. Immunized dogs rapidly cleared the wild-type pathogen infection and tested positive for bacteremia less frequently than unvaccinated controls. While immune responses varied among dogs, vaccinees consistently mounted IgG and CD4+ T-cell responses specific to E. chaffeensis throughout the assessment period. Our findings demonstrate that MLAV-mediated immune protection persists for at least one year against wild-type bacterial infection, marking a major advancement in combating this serious tick-borne disease. The data presented here serve as the foundation for further studies, elucidating the molecular mechanisms underlying virulence and vaccine development and aiding in preventing the diseases caused by E. chaffeensis and other tick-borne rickettsial pathogens.

Identification of Rickettsia spp. in Ticks Removed from Tick-Bitten Humans in Northwestern Spain.

Tick-borne rickettsioses (TBRs) are distributed worldwide and are recognized as important emerging vector-borne zoonotic diseases in Europe. The aim of this study was to identify tick-associated Rickettsia among ticks removed from humans, and to track how tick populations and their associated pathogens have changed over the years. For this purpose, we conducted a tick surveillance study in northwestern Spain between 2018 and 2022. Ticks were morphologically identified and analyzed for the presence of rickettsial pathogens through the amplification of the citrate synthase (gltA) and the outer membrane protein A (ompA) genes. PCR products were sequenced and subjected to phylogenetic analyses. We collected 7397 ticks, with Ixodes ricinus being the species most frequently isolated. Based on the PCR results, Rickettsia DNA was detected in 1177 (15.91%) ticks, and 10 members of Rickettsia were identified: R. aeschlimannii, R. conorii subsp. conorii, R. conorii subsp. raoultii, R. massiliae, R. monacensis, R. sibirica subsp. mongolitimonae, R. slovaca, R. helvetica, Candidatus R. barbariae, and Candidatus R. rioja. Some of these Rickettsia have gone previously undetected in the study region. There is clear geographic and seasonal expansion not only of tick populations, but also of the associated Rickettsia. The comparison of our data with those obtained years ago provides a clear idea of how the spatiotemporal distributions of ticks and their associated Rickettsiae have changed over the years.

Dog ectoparasites as sentinels for pathogenic Rickettsia and Bartonella in rural Guatemala.

Fleas and ticks serve as vectors of multiple pathogens in the genera Rickettsia and Bartonellathat cause diseases in humans and other animals. Although human rickettsiosis and bartonellosis have been reported in all countries in Central America, limited research has been conducted to investigate the natural cycles of flea- and tick-borne rickettsiosis and bartonellosis, especially in Guatemala. We evaluated dog parasites as sentinels for zoonotic disease risk in rural Guatemala by sampling ticks and fleas from dogs, which were then identified and individually screened for Rickettsia and Bartonella. A total of 77 households were surveyed and 80.52% of them had dogs. Overall, 133 dogs were examined for fleas and ticks, of which 68.42% had fleas and 35.34% had ticks. A total of 433 fleas and 181 ticks were collected from the infested dogs, with an additional 33 ticks collected from house walls. Three flea species were identified: Ctenocephalides felis (70%), Echidnophaga gallinacea(11.8%), and Pulex sp. (17.8%). Among the collected ticks, 97% were Rhipicephalus sanguineus with the rest being Amyblyomma cajennense, A. auricularium, and A. ovale. Rickettsia felis were detected in six C. felis, in one Pulex sp., and in two R. sanguineus, while Candidatus R. senegalensis was detected in one C. felis. Bartonella was detected only in fleas, including three Pulexsp. infected with B. vinsonii subsp. Berkhoffii, B. henselae, and Bartonella sp., respectively, and 11 C. felis infected with B. henselae. This study reports Candidatus R. senegalensis and B. vinsonii subsp. Berkhoffiiin Guatemala for the first time, and indicates the potential risk of human and dog exposure to Rickettsia and Bartonella species. These results show that dogs provide critical information relevant to managing human potential exposure to flea- and tick-borne pathogens in rural Guatemala.

Newly Recognized Spotted Fever Group Rickettsia as Cause of Severe Rocky Mountain Spotted Fever-Like Illness, Northern California, USA.

The incidence of spotted fever group (SFG) rickettsioses in the United States has tripled since 2010. Rocky Mountain spotted fever, the most severe SFG rickettsiosis, is caused by Rickettsia rickettsii. The lack of species-specific confirmatory testing obfuscates the relative contribution of R. rickettsii and other SFG Rickettsia to this increase. We report a newly recognized rickettsial pathogen, Rickettsia sp. CA6269, as the cause of severe Rocky Mountain spotted fever-like illness in 2 case-patients residing in northern California. Multilocus sequence typing supported the recognition of this pathogen as a novel Rickettsia genotype most closely related to R. rickettsii. Cross-reactivity observed for an established molecular diagnostic test indicated that Rickettsia sp. CA6269 might be misidentified as R. rickettsii. We developed a Rickettsia sp. CA6269-specific real-time PCR to help resolve this diagnostic challenge and better characterize the spectrum of clinical disease and ecologic epidemiology of this pathogen.

Diversity of Rickettsia species in collected ticks from Southeast Iran

Rickettsia occurs worldwide and rickettsiosis is recognized as an emerging infection in several parts of the world. Ticks are reservoir hosts for pathogenic Rickettsia species in humans and domestic animals. Most pathogenic Rickettsia species belong to the spotted Fever Group (SFG). This study aimed to identify and diagnose tick fauna and investigate the prevalence of Rickettsia spp. in ticks collected from domestic animals and dogs in the rural regions of Kerman Province, Southeast Iran. In this study, tick species (fauna) were identified and 2100 ticks (350 pooled samples) from two genera and species including Rhipicephalus linnaei (1128) and Hyalomma deteritum (972) were tested to detect Rickettsia genus using Real-time PCR. The presence of the Rickettsia genus was observed in 24.9% (95%CI 20.28–29.52) of the pooled samples. Sequencing and phylogenetic analyses revealed the presence of Rickettsia aeschlimannii (48.98%), Rickettsia conorii israelensis (28.57%), Rickettsia sibirica (20.41%), and Rickettsia helvetica (2.04%) in the positive samples. The results showed a significant association between county variables and the following variables: tick spp. (p < 0.001), Rickettsia genus infection in ticks (p < 0.001) and Rickettsia spp. (p < 0.001). In addition, there was a significant association between tick species and host animals (dogs and domestic animals) (p < 0.001), Rickettsia spp infection in ticks (p < 0.001), and Rickettsia spp. (p < 0.001). This study indicates a high prevalence of Rickettsia spp. (SFG) in ticks of domestic animals and dogs in rural areas of Kerman Province. The health system should be informed of the possibility of rickettsiosis and the circulating species of Rickettsia in these areas.

Sludge biolysis pretreatment to reduce antibiotic resistance genes (ARGs): Insight into the relationship between potential ARGs hosts and BALOs' preferred prey

As an important reservoir of antibiotic resistance genes (ARGs), the sludge discharged from wastewater treatment plants is the key intermediate for ARG transport into the environment. Bdellovibrio-and-like organisms (BALOs) are predatory bacteria that are expected to attack antibiotic-resistant bacteria (ARB). In this study, the screened BALOs (C3 & D15) were mixed with the sludge for biolysis to achieve the satisfying removal efficiencies of six tet genes, two sul genes, and one mobile genetic element (intl 1). Among them, tet(Q) demonstrated the highest reduction rate in relative abundance at 87.3 ± 1.0 %, while tet(X) displayed the lowest of 11.7 ± 0.2 %. The microorganisms, including Longilinea, Methanobacterium, Acetobacterium, Sulfurimonas, allobaculum, Gaiella, AAP99, Ellin6067, Rhodoferax, Ferruginibacter and Thermomonas, were expected to play a dual role in the reduction of ARGs by serving as ARB and BALOs’ preferred prey. Meanwhile, BALOs consortium improved ARGs reduction efficiency via the expansion of the prey profile. Additionally, BALOs decreased the relative abundance of not only pathogens (Shinella, Rickettsia, Burkholderia, Acinetobacter, Aeromonas, Clostridium, Klebsiella and Pseudomonas), but also the ARGs' host pathogens (Mycobacterium, Plesiocystis, Burkholderia, and Bacteroides). Therefore, the application of BALOs for sludge biolysis are promising to decrease the sludge's public health risks via limiting the spread of ARGs and pathogens into the environment.