Canine monocytic ehrlichiosis (CME) is a highly infectious disease with zoonotic potential. Ehrlichia canis, the causative agent, is primarily transmitted by Rhipicephalus sanguineus. Tick infestation and tick-borne diseases (TBDs) are serious human and veterinary health problems due to the lack of efficient control measures. This review aims to provide information about CME, detailing epidemiology, pathogenesis, clinical manifestations, and current strategies for diagnosis, treatment, and prevention. The review delves into the biology of R. sanguineus, exploring its lifecycle, habitat, and mechanisms of E. canis transmission. Ehrlichia canis infection follows a three-phase transition: acute, subclinical, and chronic, with distinct clinical signs, from mild to severe and even life-threatening in some cases, with the potential for asymptomatic long-term carriers acting as reservoirs. Pathogenesis involves complex interactions between the pathogen and host immune responses, influencing disease severity and progression. Additionally, the review highlights challenges in controlling CME due to the widespread distribution of R. sanguineus. Genetic diversity within R. sanguineus populations and their varying vector competence further complicate control efforts. The role of environmental changes in tick distribution aligns the control of CME within the One Health concept, integrating approaches to improve outcomes for humans, animals, and the environment. Preventive measures on tick control and potential vaccine development are also reviewed.

In a world facing the unprecedented threat of antibiotic-resistant bacteria, targeted approaches to control colonization and prevent disease caused by common pathobionts offer a promising solution. Streptococcus pneumoniae (pneumococcus) is a leading cause of infections worldwide, affecting both children and adults despite available antimicrobials and vaccines. Colonization, which occurs in the form of a biofilm in the upper respiratory tract, is frequent and a prerequisite for disease and transmission. The use of live bacterial strains as biotherapeutics for infectious diseases is actively being explored. Here, we investigated the potential of commensal streptococci to control S. pneumoniae . Screening of over 300 human isolates led to the identification of seven strains (one Streptococcus oralis and six Streptococcus mitis , designated A22 to G22) with inhibitory activity against S. pneumoniae of multiple serotypes and genotypes. Characterization of A22 to G22 cell-free supernatants indicated the involvement of secreted proteins or peptides in the inhibitory effect of all S. mitis isolates. Genome analyses revealed the presence of 64 bacteriocin loci, encoding 70 putative bacteriocins, several of which are novel and absent or rare in over 7,000 publicly available pneumococcal genomes. Deletion mutants indicated that bacteriocins partially or completely explained the anti-pneumococcal activity of the commensal strains. Importantly, strains A22 to G22 were further able to prevent and disrupt pneumococcal biofilms, a proxy for nasopharyngeal colonization. These results highlight the intricacy of the interactions among nasopharyngeal colonizers and support the potential of strains A22 to G22 to be used as live biotherapeutics, alone or in combination, to control S. pneumoniae colonization. IMPORTANCE: Streptococcus pneumoniae (pneumococcus) infections remain a major public health issue despite the use of vaccines and antibiotics. Pneumococci asymptomatically colonize the human upper respiratory tract, a niche shared with several commensal Streptococcus species. Competition for space and nutrients among species sharing the same niche is well documented and tends to be more intense among closely related species. Based on this rationale, a screening of several commensal streptococci isolated from the human upper respiratory tract led to the identification of strains of Streptococcus mitis and Streptococcus oralis capable of inhibiting most pneumococcal strains, across diverse serotypes and genotypes. This inhibition was partially or wholly linked to the expression of novel bacteriocins. The selected S. mitis and S. oralis strains significantly disrupted pneumococcal biofilms, indicating a potential for using commensals as biotherapeutics to control pneumococcal colonization, a key step in preventing disease and transmission.

Parasitic diseases are an important worldwide problem threatening human health and affect millions of people. Acute diarrhea, intestinal bleeding, malabsorption of nutrients and nutritional deficiency are some of the issues related to intestinal parasitic infections. Parasites are experts in subvert the host immune system through different kinds of mechanisms. There are evidences that extracellular vesicles (EVs) have an important role in dissemination of the disease and in modulating the host immune system. Released by almost all types of cells, these nanovesicles are a natural secretory product containing multiple components of interest. The EVs are classified as apoptotic bodies, microvesicles, exosomes, ectosomes, and microparticles, according to their physical characteristics, biochemical composition and cell of origin. Interestingly, EVs play an important role in intercellular communication between parasites as well as with the host cells. Concerning Giardia lamblia, it is known that this parasite release EVs during it life cycle that modulate the parasite growth and adherence as well the immune system of the host. Here we review the recently updates on protozoa EVs, with particular emphasis on the role of EVs released by the flagellate protozoa G. lamblia in cellular communication and its potential for future applications as vaccine, therapeutic agent, drug delivery system and as diagnostic or prognostic biomarker.

Extracellular vesicles (EVs) (exossomes, microvesicles and apoptotic bodies) have been well acknowledged as mediators of intercellular communications in prokaryotes and eukaryotes. Lipids are essential molecular components of EVs but at the moment the knowledge about the lipid composition and the function of lipids in EVs is limited and as for now none lipidomic studies in Giardia EVs was described. Therefore, the focus of the current study was to conduct, for the first time, the characterization of the polar lipidome, namely phospholipid and sphingolipid profiles of G. lamblia trophozoites, microvesicles (MVs) and exosomes, using C18—Liquid Chromatography—Mass Spectrometry (C18-LC-MS) and Tandem Mass Spectrometry (MS/MS). A total of 162 lipid species were identified and semi-quantified, in the trophozoites, or in the MVs and exosomes belonging to 8 lipid classes, including the phospholipid classes phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), cardiolipins (CL), the sphingolipid classes sphingomyelin (SM) and ceramides (Cer), and cholesterol (ST), and 3 lipid subclasses that include lyso PC (LPC), lyso PE (LPE) and lyso PG (LPG), but showing different abundances. This work also identified, for the first time, in G. lamblia trophozoites, the lipid classes CL, Cer and ST and subclasses of LPC, LPE and LPG. Univariate and multivariate analysis showed clear discrimination of lipid profiles between trophozoite, exosomes and MVs. The principal component analysis (PCA) plot of the lipidomics dataset showed clear discrimination between the three groups. Future studies focused on the composition and functional properties of Giardia EVs may prove crucial to understand the role of lipids in host-parasite communication, and to identify new targets that could be exploited to develop novel classes of drugs to treat giardiasis.

Objectives: This study aimed to investigated the role of Giardia extracellular vesicles (EVs) in intercellular communication and to evaluated their potential as vaccine candidates. Methods: The immunomodulatory effects of Giardia EVs were assessed in mouse macrophages and human monocyte-derived dendritic cells (Mo-DCs), with a particular focus on key inflammatory signaling pathways. In vivo immunogenicity was evaluated following EV administration, and the antigenic composition of EV cargo was characterized by proteomic analysis. Results: Giardia EVs activated pro-inflammatory signaling pathways in mouse macrphages, including SAPK/JNK, ERK1/2, and NF-κB. This activation was associated with IκB-α degradation and nuclear translocation of p65. Furthermore, EV stimulation significantly upregulated the expression of pro-inflammatory genes, including Il1β, Il6, Il4, Ptgs2, Nos2, and Tnf, with log₂ fold changes ranging from 3.9 to 15.8. Consistently, EVs increased iNOS protein expression (28–45%) and nitrite production (9.6–12.3-fold). In human Mo-DCs, Giardia EVs promoted cellular maturation, as evidenced by increased expression of MHC-II, CD80, and CD86, and enhanced T-cell proliferation with a Th1-skewed profile. In vivo immunization induced antigen-specific antibody responses, with IgG subclass distribution indicative of a balanced Th1/Th2 response. Proteomic analysis identified immunoreactive EV-associated proteins, including elongation factor 1-alpha, α-7.3 giardin, tubulin, and variant surface proteins (VSPs), which are well-established antigens in Giardia infection, with prominent bands observed at approximately 22 kDa and 50 kDa. Conclusions: Collectively, these findings demonstrate that Giardia EVs modulate innate immune responses in vitro, elicit antigen-specific humoral immunity in vivo, and contain conserved immunogenic proteins. These properties support their potential as a promising cell-free vaccine platform against giardiasis.

In this study, we investigated the role of Giardia EVs in cellular communication and their potential as vaccine candidates. Our findings revealed that Giardia EVs activate pro-inflammatory signalling cascades, including SAPK/JNK and ERK1/ERK2, as well as the NF-kB pathway, resulting in IκB-α degradation and p65 translocation to the nucleus, in mouse macrophages. Moreover, Giardia EVs increased the expression of genes encoding pro-inflammatory molecules, such as Il1β, Il6, Il4, Ptgs2, Nos2, and Tnf. Interestingly, Giardia EVs enhanced the maturation status of human Mo-DCs and significantly increased T-cell proliferation with a Th1 profile. Immunization studies demonstrated that Giardia EVs elicited antigen-specific antibodies, with IgG subclasses indicating a balance Th1/ Th2 response. Mass spectrometry analysis identified EV proteins (22 KDa and 50 KDa) that bind to serum antibodies of immunized mice including elongation factor 1-alpha, Alpha-7.3 giardin, tubulin, and Variant Surface Proteins (VSP), known antigenic proteins in Giardia infections. Overall, our results indicated that Giardia EVs modulate innate immune cells in vitro, induce antibody-based immune response in vivo, and contain conserved immunogenic proteins. Consequently, Giardia EVs hold promise as a cell-free vaccine candidate for giardiasis.