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Pathogenic Rickettsia species target vascular endothelial cells and cause systemic vasculitis. As obligate intracellular bacterial pathogens, Rickettsia must secure nutritional resources within the cytoplasm of endothelial cells while simultaneously subverting the innate immune defense system. With advances in rickettsial and host genetics, recent studies have identified novel molecular mechanisms involved in the complex interactions between Rickettsia and endothelial cells. However, it remains unclear how Rickettsia shields pathogen-derived immune stimulants, such as lipopolysaccharides (LPS) and peptidoglycan fragments, from immune recognition during intracellular replication. Prior work described two Rickettsia conorii variants with kkaebi transposon insertions in the polysaccharide synthesis operon ( pso ). Biochemical and immunological analyses revealed that pso is responsible for the biosynthesis of O-antigen (O-Ag) and the proper assembly of surface proteins. In the present work, we document that pso variant HK2 exhibits reduced capacities to adhere to and invade microvascular endothelial cells. Despite the low intracellular abundance, HK2 induced significantly higher levels of proinflammatory cytokines and chemokines, leading to premature cell death. Notably, HK2 exhibited defective intracellular survival in bone marrow-derived macrophages. This inability to dampen endothelial cell-mediated immune stimulation and resist macrophage-induced bactericidal activities resulted in the rapid elimination of viable Rickettsia in the mouse model of spotted fever. Further, when tested as a live-attenuated vaccine, HK2 elicited robust protective immunity against lethal spotted fever pathogenesis. Our work highlights the crucial role of pso in enabling Rickettsia to evade immune surveillance during intracellular replication within endothelial cells, ultimately delaying pathogen-induced programmed cell death and escaping immune defense mechanisms.

A total of 68 adult hard ticks were recovered from 5 cases of infestation involving pet rabbits (Oryctolagus cuniculus domesticus) in Malaysia. The ticks were later identified as the tropical brown dog tick, Rhipicephalus linnaei (Audouin, 1826). Pathogen screening showed 2 out of the 68 Rh. linnaei ticks (2.94% infection rate) were positive for Rickettsia. BLAST analyses revealed that both samples were identical to Rickettsia conorii subsp. raoultii. This paper reports the first detection of R. conorii subsp. raoultii in Rh. linnaei, also highlighting the potential zoonotic threat posed by ticks infesting exotic pets such as rabbits.

Rickettsia species are an important cause of emerging infectious diseases in people and animals, and rickettsiosis is one of the oldest known vector-borne diseases. Laboratory diagnosis of Rickettsia is complex and time-consuming. This study was aimed at developing two quantitative real-time PCRs targeting ompB and ompA genes for the detection, respectively, of Rickettsia spp. and R. conorii DNA. Primers were designed following an analysis of Rickettsia gene sequences. The assays were optimized using SYBR Green and TaqMan methods and tested for sensitivity and specificity. This study allowed the development of powerful diagnostic methods, able to detect and quantify Rickettsia spp. DNA and differentiate R. conorii species.

Hamadryas baboons (Papio hamadryas) may transmit zoonotic vector-borne pathogens to visitors and workers frequenting zoological parks. We molecularly screened 33 baboons for vector-borne pathogens. Three (9.1%) of 33 animals tested positive for Rickettsia conorii subspecies israelensis. Clinicians should be aware of potential health risks from spatial overlapping between baboons and humans.

Summary Rickettsia conorii, a member of the spotted fever group (SFG) of the genus Rickettsia and causative agent of Mediterranean spotted fever, is an obligate intracellular pathogen capable of infecting various mammalian cell types. SFG rickettsiae express two major immunodominant surface cell antigen (Sca) proteins, OmpB (Sca5) and OmpA (Sca0). While OmpB‐mediated entry has been characterized, the contribution of OmpA has not been well defined. Here we show OmpA expression in Escherichia coli is sufficient to mediate adherence to and invasion of non‐phagocytic human endothelial cells. A recombinant soluble C‐terminal OmpA protein domain (954–1735) with predicted structural homology to the Bordetella pertussis pertactin protein binds mammalian cells and perturbs R. conorii invasion by interacting with several mammalian proteins including β1 integrin. Using functional blocking antibodies, small interfering RNA transfection, and mouse embryonic fibroblast cell lines, we illustrate the contribution of α2β1 integrin as a mammalian ligand involved in R. conorii invasion of primary endothelial cells. We further demonstrate that OmpA‐mediated attachment to mammalian cells is in part dependent on a conserved non‐continuous RGD motif present in a predicted C‐terminal ‘pertactin’ domain in OmpA.Our results demonstrate that multiple adhesin–receptor pairs are sufficient in mediating efficient bacterial invasion of R. conorii.

A fatal case of Mediterranean spotted fever associated with septic shock was reported in a 61-year-old man living in a village in southeastern Iran. The patient had a history of tick bite a few days before symptom onset. Phylogenetic analysis confirmed infection by Rickettsia conorii subspecies israelensis.

Small regulatory RNAs (sRNAs) are now widely recognized for their role in the post-transcriptional regulation of bacterial virulence and growth. We have previously demonstrated the biogenesis and differential expression of several sRNAs in Rickettsia conorii during interactions with the human host and arthropod vector, as well as the in vitro binding of Rickettsia conorii sRNA Rc_sR42 to bicistronic cytochrome bd ubiquinol oxidase subunits I and II (cydAB) mRNA. However, the mechanism of regulation and the effect of sRNA binding on the stability of the cydAB bicistronic transcript and the expression of the cydA and cydB genes are still unknown. In this study, we determined the expression dynamics of Rc_sR42 and its cognate target genes, cydA and cydB, in mouse lung and brain tissues during R. conorii infection in vivo and employed fluorescent and reporter assays to decode the role of sRNA in regulating cognate gene transcripts. Quantitative RT-PCR revealed significant changes in the expression of sRNA and its cognate target gene transcripts during R. conorii infection in vivo, and a greater abundance of these transcripts was observed in the lungs compared to brain tissue. Interestingly, while Rc_sR42 and cydA exhibited similar patterns of change in their expression, indicating the influence of sRNA on the mRNA target, the expression of cydB was independent of sRNA expression. Further, we constructed reporter plasmids of sRNA and cydAB bicistronic mRNA to decipher the role of sRNA on CydA and CydB expression. We observed increased expression of CydA in the presence of sRNA but detected no change in CydB expression in the presence or absence of sRNA. In sum, our results demonstrate that the binding of Rc_sR42 is required for the regulation of cydA but not cydB. Further studies on understanding the influence of this interaction on the mammalian host and tick vector during R. conorii infection are in progress.

Vector-borne diseases, particularly those caused by Rickettsia species, pose a significant public health threat in Europe. Despite extensive research on tick-borne pathogens in various European countries, Luxembourg has yet not been studied for Rickettsia spp. in ticks infesting domestic animals. This study aimed to fill this gap by investigating the presence of Rickettsia spp. in Ixodes ricinus ticks collected from domestic dogs in Luxembourg between April 2023 and April 2024. A total of 61 ticks were examined using molecular techniques, including PCR amplification of the outer membrane protein B (ompB), outer membrane protein A (ompA), and citrate synthase (gltA) genes. Results revealed the presence of R. helvetica and R. conorii subsp. raoultii, with 4.9% of ticks testing positive for Rickettsia spp. Phylogenetic analysis confirmed the high genetic identity of the sequences obtained with previously described strains from Europe and Asia. This study highlights the potential risk of emerging tick-borne diseases in Luxembourg and emphasizes the need for ongoing surveillance to better understand the spread of Rickettsia spp. in Europe, particularly as climate change may facilitate the expansion of tick populations and their associated pathogens.

ABSTRACT Rickettsia species are Gram-negative, pleomorphic coccobacilli that are obligate intracellular pathogens transmitted by arthropod vectors such as ticks. Among them, Rickettsia conorii, the causative agent of Mediterranean spotted fever (MSF), is endemic in many Mediterranean countries, including Turkey. This case series describes three patients from Balıkesir, Turkey, who developed high-grade fever, generalized maculopapular rash involving the palms and feet soles, arthralgia, and necrotic eschars (tache noire) at the tick bite sites. All cases occurred during summer and had documented exposure to Rhipicephalus sanguineus. Laboratory evaluations ruled out other tick-borne diseases, while real-time PCR performed on skin biopsy samples confirmed Rickettsia spp. Subsequent DNA sequencing of the gltA and ompA gene regions enabled species identification. Additionally, serological tests showed a significant rise in IgM and IgG antibody titers reacting with Rickettsia conorii antigen by indirect immunofluorescence assay. All patients were treated with doxycycline and recovered without complications. This case series highlights the importance of considering rickettsial infections in the differential diagnosis of febrile patients with rash and recent tick exposure, especially in endemic regions during warm seasons.

Ticks are important vectors for Rickettsia spp. of the spotted fever group all around the world. Rickettsia conorii is the etiological agent of boutonneuse fever in the Mediterranean region and Africa. Tick identification was based on morphological features and further characterized using the 16S rRNA gene. The ticks were individually tested using pan- Rickettsia real-time-PCR for screening, and 23S-5S intergenic spacer region, 16S rDNA, glt A, sca 4, omp B, and ompA genes were used to analyze the Rickettsia positive samples . Rickettsia conorii ssp. caspia was detected in tick collected in Zambia for the first time, thus demonstrating the possibility of the occurrence of human disease, namely Astrakhan fever, due to this Rickettsia ssp. in this region of Africa. The prevalence of R. conorii ssp. caspia was 0.06% (one positive tick out of 1465 tested ticks) and 0.07% (one positive tick out of 1254 tested Rh. sanguineus ).