Given the proven zoonotic potential of the intestinal protozoan Blastocystis sp., a fast-growing number of surveys are being conducted to identify potential animal reservoirs for transmission of the parasite. Nevertheless, few epidemiological studies have been conducted on farmed animals in Egypt. Therefore, a total of 1089 fecal samples were collected from herbivores (sheep, goats, camels, horses, and rabbits) in six Egyptian governorates (Dakahlia, Gharbia, Kafr El Sheikh, Giza, Aswan, and Sharqia). Samples were screened for the presence of Blastocystis sp. by real-time PCR followed by sequencing of positive PCR products and phylogenetic analysis for subtyping of the isolates. Overall, Blastocystis sp. was identified in 37.6% of the samples, with significant differences in frequency between animal groups (sheep, 65.5%; camels, 62.2%; goats, 36.0%; rabbits, 10.1%; horses, 3.3%). Mixed infections were reported in 35.7% of the Blastocystis sp.-positive samples. A wide range of subtypes (STs) with varying frequency were identified from single infections in ruminants including sheep (ST1–ST3, ST5, ST10, ST14, ST21, ST24, ST26, and ST40), goats (ST1, ST3, ST5, ST10, ST26, ST40, ST43, and ST44), and camels (ST3, ST10, ST21, ST24–ST26, ST30, and ST44). Most of them overlapped across these animal groups, highlighting their adaptation to ruminant hosts. In other herbivores, only three and two STs were evidenced in rabbits (ST1–ST3) and horses (ST3 and ST44), respectively. The greater occurrence and wider genetic diversity of parasite isolates among ruminants, in contrast to other herbivores, strongly suggested that dietary habits likely played a significant role in influencing both the colonization rates of Blastocystis sp. and ST preference. Of all the isolates subtyped herein, 66.3% were reported as potentially zoonotic, emphasizing the significant role these animal groups may play in transmitting the parasite to humans. These findings also expand our knowledge on the prevalence, genetic diversity, host specificity, and zoonotic potential of Blastocystis sp. in herbivores.

Blastocystis sp. is the most common single-celled eukaryote colonizing the human gastrointestinal tract worldwide. Because of the proven zoonotic potential of this protozoan, sustained research is therefore focused on identifying various reservoirs of transmission to humans, and in particular animal sources. Numerous groups of animals are considered to be such reservoirs due to their handling or consumption. However, some of them, including mollusks, remain underexplored. Therefore, a molecular epidemiological survey conducted in wild mussels was carried out in Northern France (Hauts-de-France region) to evaluate the frequency and subtypes (STs) distribution of Blastocystis sp. in these bivalve mollusks. For this purpose, 100 mussels (Mytilus edulis) were randomly collected in two sampling sites (Wimereux and Dannes) located in the vicinity of Boulogne-sur-Mer. The gills and gastrointestinal tract of each mussel were screened for the presence of Blastocystis sp. by real-time polymerase chain reaction (qPCR) assay followed by direct sequencing of positive PCR products and subtyping through phylogenetic analysis. In parallel, sequences of potential representative Blastocystis sp. isolates that were previously obtained from temporal surveys of seawater samples at marine stations offshore of Wimereux were integrated in the present analysis. By taking into account the qPCR results from all mussels, the overall prevalence of the parasite was shown to reach 62.0%. In total, more than 55% of the positive samples presented mixed infections. In the remaining mussel samples with a single sequence, various STs including ST3, ST7, ST14, ST23, ST26 and ST44 were reported with varying frequencies. Such distribution of STs coupled with the absence of a predominant ST specific to these bivalves strongly suggested that mussels might not be natural hosts of Blastocystis sp. and might rather be carriers of parasite isolates from both human and animal (bovid and birds) waste. These data from mussels together with the molecular identification of isolates from marine stations were subsequently discussed along with the local geographical context in order to clarify the circulation of this protozoan in this area. The identification of human and animal STs of Blastocystis sp. in mussels emphasized the active circulation of this protozoan in mollusks and suggested a significant environmental contamination of fecal origin. This study has provided new insights into the host/carrier range and transmission of Blastocystis sp. and emphasized its potential as an effective sentinel species for water quality and environmental contamination.

One hundred years after planctomycetes were discovered and 50 years since the first isolate was successfully cultured, this bacterial phylum remains enigmatic in many ways. In the last few decades, a significant effort to characterize new isolates has resulted in >150 described species, allowing a more comprehensive analysis of their features. However, metagenomic studies reveal that a diverse group of planctomycetes has yet to be cultured and characterized, and that many biological surprises are yet to be revealed. This is the case for the recently discovered phagotrophic Candidatus Uabimicrobium, which challenges our understanding of the distinction between prokaryotes and eukaryotes. The unique biology of planctomycete cells, such as their ability to divide without the FtsZ protein, their complex structure and characteristic morphology, their relatively large genomes containing many genes with unknown function, and their variable metabolic capabilities, imposes significant barriers for researchers. Although ubiquitous, the precise ecological roles of planctomycetes in various environments are still not fully understood. However, their distinctive metabolism opens the door to a large number of potential biotechnological applications, which are beginning to be unveiled. In this article, we first review the historical milestones in planctomycetes research and describe the pioneers of the field. We then describe the controversies and their resolutions, we highlight the past discoveries and current interrogations related to planctomycetes, and discuss the ongoing challenges that hinder a comprehensive understanding of their biology. We end up with directions for exploring the biology and ecological roles of these fascinating organisms.

This study is the first to report Blastocystis sp. in dairy cattle from Terceira Island, part of the Azores Archipelago. Blastocystis sp. is an enteric protozoan with high genetic diversity and is associated with both zoonotic and non-zoonotic subtypes (STs). The present survey aimed to fill the gap in knowledge regarding the occurrence and genetic characterization of Blastocystis sp. isolates in Holstein-Friesian cattle in this geographical area. A total of 116 stool samples were thus collected from dairy cows, and Blastocystis sp. DNA was detected using qPCR. The results revealed a 14.7% occurrence rate (17/116), with seven STs identified including ST3, ST5, ST7, ST10, ST14, ST25, and ST42, with variable frequency. Some of these STs (ST5, ST7, ST10, and ST14) are potentially zoonotic, underscoring their potential public health significance. However, the low frequency of Blastocystis sp. in this animal cohort possibly attributed to the extensive grazing system practiced on Terceira Island and the limited presence of zoonotic isolates, suggests a minimal risk of transmission from cattle to humans. This study highlights the importance of further research on the transmission dynamics of Blastocystis sp., particularly in regions with varying farming practices, to better understand its epidemiology and zoonotic potential.

Copper (Cu) is an essential micronutrient that serves as a cofactor for many enzymes but becomes toxic when present in excess. In most bacteria, CopA-like P1B-type ATPases mediate Cu detoxification by exporting cytoplasmic Cu to the periplasm or extracellular environment. In this study, we show that Caulobacter vibrioides lacks a canonical CopA-like ATPase but encodes a single FixI/CcoI-type Cu-transporting ATPase, previously implicated in Cu delivery to the cbb₃-type cytochrome c oxidase (Cox) in species such as Rhodobacter capsulatus. C. vibrioides harbors two terminal cytochrome c oxidases in its cytoplasmic membrane: an aa₃-type and a cbb₃-type Cox. We also demonstrate that the activity of cbb₃-Cox requires the FixI-type Cu transporter and the periplasmic Cu chaperone PccA. In contrast, aa₃-Cox activity depends on PccA and the inner membrane-bound protein CtaG. Since the mechanism of Cu acquisition for aa₃-Cox remains largely unknown, we conducted a genetic screen and identified a novel outer membrane TonB-dependent receptor (TccA) that is specifically required for aa₃-Cox function. We also showed that cbb₃-Cox is upregulated under microaerobic conditions, possibly such as those encountered on solid media where O₂ diffusion is limited. Under normoxic conditions, the expression and the activity of cbb₃-Cox decrease, and aa₃-Cox becomes the predominant terminal oxidase. These findings demonstrate that C. vibrioides differentially utilizes its Cox enzymes in response to O₂ availability and relies on a distinct Cu trafficking pathway for their maturation, including an outer membrane component that has not been previously described in bacterial Cu homeostasis.

Reconstructing the Tree of Life (ToL) remains a central challenge in evolutionary biology. While sequence-based methods have resolved much of life’s diversity, deep evolutionary relationships often remain elusive, particularly across the three superkingdoms of life. Here, we present a complementary approach using genome-wide occurrence of protein domain combinations as phylogenetic signal. We analyzed 5,343 complete proteomes spanning Bacteria, Archaea, and Eukaryota, and inferred a pan-genomic ToL from ∼77,000 domain combinations, including isolated domains and domain pairs. Our reconstruction recovers major clades and captures both vertical inheritance and signals from non-vertical processes such as lateral gene transfer, endosymbiosis, and genome reduction. Ancestral reconstructions reveal functional innovations that mark key evolutionary transitions—such as the emergence of vesicle trafficking in Eukaryota or methanogenesis in Archaea. We found that organisms with reduced genomes, like Chlamydiota, tend to be misplaced in domain combination-based phylogenies due to sparse combinatorial data. Our results demonstrate that domain combinations carry a strong, evolutionary signal that is both biologically and functionally informative. While not a substitute for sequence-based phylogenetics, domain combination-based trees offer an alternative perspective, capable of integrating both vertical and lateral processes into a unified evolutionary framework. Author Summary: Traditional evolutionary trees are typically built from sequences, but this approach struggles to resolve deep branches in the Tree of Life. Here, we use combinations of protein domains—the functional modules that make up proteins to resolve major relationships in the tree. By analyzing over 5,000 complete proteomes from Bacteria, Archaea, and Eukaryotes, we reconstruct a Tree of Life that reflects both vertical inheritance and key evolutionary processes like endosymbiosis and gene transfer. This domain-based approach not only recovers known relationships but also reveals how major biological innovations emerged. Our results show that domain combinations provide a powerful and interpretable signal for studying evolution at a genomic scale.