Parabasalids, formally classified as the phylum Parabasalia, are a diverse group of anaerobic or microaerophilic unicellular eukaryotes that lack typical mitochondria but possess hydrogenosomes—double-membrane-bound organelles derived from mitochondria that generate energy via hydrogen production under low-oxygen conditions.[1] These protists are distinguished by a unique cytoskeletal feature called the parabasal body, which anchors their multiflagellar apparatus and supports closed mitosis known as pleuromitosis, along with forms ranging from flagellates to amoeboflagellates.[1] Primarily symbiotic in the hindguts of wood-feeding insects such as termites and cockroaches, parabasalids play a crucial role in lignocellulose digestion, enabling host nutrient acquisition from plant material, while a few species are free-living or parasitic.[2]The phylum encompasses over 400 described species, with molecular analyses of small subunit ribosomal RNA (SSU rRNA) sequences revealing even greater diversity, including hundreds of environmental lineages.[1] Recent taxonomic revisions, based on integrated ultrastructural, molecular phylogenetic, and ecological data, divide Parabasalia into 11 classes, 16 orders, and 31 families, refining earlier schemes and highlighting monophyly supported by shared traits like the parabasal filament.[1] Major classes include Trichomonadea, which contains small, simple trichomonads like Trichomonas vaginalis—the causative agent of human trichomoniasis, a prevalent sexually transmitted infection—and Trichonymphea, featuring large, multiflagellated hypermastigotes such as Trichonympha species that dominate termite guts and exhibit complex symbiotic associations with bacteria.[3][1] Other notable groups, like Cristamonadea and Spirotrichonymphea, include amoeboid or spirochete-associated forms, such as Mixotricha paradoxa, which integrates bacterial ectosymbionts for motility and wood degradation.[1]Parabasalids are among the earliest-diverging eukaryotic lineages,[2] and their hydrogenosome-based metabolism reflects adaptations to anoxic environments. In termite symbiosis, these protists not only hydrolyze cellulose using endogenous enzymes but also produce hydrogen that fuels acetogenic bacteria, contributing to the host's energy needs and underscoring their ecological significance in carbon cycling within forest ecosystems.[4] Parasitic members, conversely, pose public health concerns; T. vaginalis infects millions annually, with an estimated 156 million new cases globally in 2020, leading to complications like increased HIV susceptibility, though it lacks a cyst stage for environmental persistence.[3][5] Ongoing research leverages genomic and transcriptomic tools to explore their reductive evolution and potential biotechnological applications, such as biohydrogen production.[1]
General Characteristics
Morphology and Ultrastructure
Parabasalids exhibit a predominantly flagellate body plan, characterized by a pear-shaped or elongated cell body ranging from 5 to over 200 μm in length, depending on the lineage. The cells possess a single nucleus and multiple flagella emerging from basal bodies organized into mastigont systems, which are complexes of basal bodies, associated fibers, and cytoskeletal elements. A defining feature is the parabasal body, a Golgi-like organelle positioned near the basal bodies and supported by distinctive parabasal fibers that connect to the nuclear envelope; this structure facilitates secretion and is unique to parabasalids among eukaryotes.[1][6]At the ultrastructural level, parabasalids lack typical mitochondria, instead featuring hydrogenosomes—double-membrane-bound organelles, typically 0.5–2 μm in diameter, that represent modified mitochondria adapted for anaerobic conditions—and a distinctive Golgi apparatus incorporated into the parabasal body. The cytoskeleton is highly specialized, comprising microtubular structures such as the pelta (a forward-projecting sheet) and axostyle (a supportive rod extending posteriorly through the cell), along with parabasal filaments that reinforce the flagellar apparatus and maintain cell shape. In trichomonads, for instance, the axostyle forms an internal skeletal rod that tapers or abruptly narrows, aiding in structural integrity and motility.[1][7][6]Morphological variations are pronounced across parabasalid lineages. Trichomonads typically have 4–6 flagella, including a recurrent flagellum that forms an undulating membrane supported by a costa (a striated fiber), enabling a characteristic wavelike motion. Hypermastigotes, in contrast, bear numerous flagella—often hundreds or thousands—arranged in longitudinal bands or rostral clusters, supporting their larger, more complex bodies adapted for symbiotic lifestyles. Some parabasalids display amoeboflagellate or fully amoeboid forms, with reduced flagella and pseudopodia-like extensions for crawling, as seen in genera like Histomonas and Dientamoeba. These adaptations highlight the group's structural diversity while retaining core ultrastructural traits like the parabasal body and hydrogenosomes.[1][6]
Metabolism and Biochemistry
Parabasalids are anaerobic or microaerophilic protists that lack typical mitochondria and instead possess hydrogenosomes, specialized organelles that facilitate anaerobic energy metabolism by converting pyruvate into ATP, hydrogen gas (H2), and acetate. This process occurs through substrate-level phosphorylation, where pyruvate is decarboxylated to acetyl-CoA by pyruvate:ferredoxin oxidoreductase (PFO), generating reduced ferredoxin as an electron carrier. The reduced ferredoxin then donates electrons to [FeFe]-hydrogenase, which reduces protons to produce H2, while acetyl-CoA is further metabolized to acetate via acetate:succinate CoA-transferase (ASCT) and succinyl-CoA synthetase (SCS), yielding additional ATP. This hydrogenosomal pathway enables parabasalids to thrive in oxygen-depleted environments, such as animal guts or urogenital tracts, without relying on oxidative phosphorylation.[8][9][10]Glycolysis in parabasalids occurs in the cytosol and supplies pyruvate to the hydrogenosomes, featuring adaptations like pyrophosphate-dependent phosphofructokinase (PPi-PFK) and pyruvate phosphate dikinase (PPDK) in species such as Tritrichomonas foetus, which help maintain energetic efficiency under anaerobic conditions.[11] Many parabasalids exhibit nutritional dependencies on host-derived lipids, particularly cholesterol and other sterols, as they possess limited capacity for de novosterolbiosynthesis and instead uptake these molecules from their environment via mechanisms involving low-density lipoprotein degradation. For instance, Trichomonas vaginalis requires exogenous cholesterol for membrane integrity and growth, incorporating it as a major lipid component.[12][13][14]Biochemical diversity exists across parabasalid lineages, with variations in hydrogenosome function and fermentation end products reflecting adaptations to different symbiotic niches. In trichomonads like Trichomonas vaginalis, the canonical acetate and H2 production predominates, but some lineages, such as those in termite guts (e.g., Trichonympha), show enhanced acetatefermentation pathways that support mutualistic hydrogen transfer to prokaryotic partners. Other parabasalids may produce additional fermentative products like ethanol alongside acetate, allowing flexibility in redox balancing and energy yield. These metabolic variations underscore the group's evolutionary adaptations to anaerobic, host-associated lifestyles.[15][16][17]
Classification
Taxonomic History
The taxonomic history of parabasalids begins in the 19th century with early microscopic observations of flagellated protists. In 1836, Alfred Donné identified Trichomonas vaginalis in human vaginal secretions, describing it as an animalcule with undulating flagella, marking the first documented encounter with a parabasalid species.[18] Initially, these organisms were classified among flagellates and often grouped with trypanosomes due to similarities in their locomotor appendages and parasitic lifestyles in vertebrate hosts.[19] Subsequent descriptions in the late 1800s and early 1900s expanded knowledge of parabasalids as symbionts in insects, particularly termites, but classifications remained rudimentary, emphasizing morphological traits like flagellar arrangement without recognizing unifying features.[20]Advancements in the 20th century centered on ultrastructural discoveries that defined parabasalids as a distinct group. In 1911, Konstanty Janicki recognized the parabasal body—a Golgi-derived structure associated with the flagellar apparatus—in flagellates from cockroaches and termites, introducing a key diagnostic trait that distinguished them from other flagellates.[21] This was followed by detailed studies of termite symbionts, leading to the establishment of the orders Trichomonadida (simpler forms with fewer flagella) and Hypermastigida (complex, multiflagellated forms) in mid-century classifications, such as those by Honigberg (1963), which formalized their separation based on mastigont complexity and host associations.[22] These morphological frameworks dominated until the late 20th century, grouping parabasalids within broader protozoan categories like Zoomastigophora.[23]The molecular era, beginning in the 1990s, revolutionized parabasalid taxonomy through small subunit ribosomal RNA (SSU rRNA) phylogenies, which placed them alongside diplomonads and retortamonads in the phylum Metamonada, highlighting their amitochondriate nature and deep eukaryotic branching.[24] The 2000s saw debates over their affiliation within the supergroup Excavata, with analyses confirming monophyly alongside euglenozoans and others based on shared cytoskeletal features and molecular markers, though long-branch attraction artifacts complicated early trees.[25] Key milestones include a 2012 PLOS One study by Noda et al., which used concatenated protein markers to resolve internal relationships and root the parabasalid tree more accurately, revealing hidden diversity.[26] Most recently, Boscaro et al.'s 2024 update revised the classification into classes such as Trichomonadea and Spirotrichonymphea, integrating genomic data to reflect evolutionary divergences while maintaining morphological congruence.[27]
Current Classification
Parabasalia is recognized as a phylum within the eukaryotic supergroup Excavata, characterized by shared traits such as hydrogenosomes, a parabasal body, and closed mitosis (pleuromitosis).[1]The current taxonomic framework, as revised by Boscaro et al. in 2024, divides the phylum into 11 classes, 16 orders, and 31 families, integrating morphological features with molecular phylogenies derived primarily from small subunit ribosomal RNA (SSU rRNA) gene sequences.[1] These classes encompass diverse lineages, broadly separable into those with simpler flagellation patterns (e.g., class Trichomonadea, featuring genera such as Trichomonas and Dientamoeba with typically 4–6 flagella) and those with complex, multiflagellate arrangements (e.g., class Spirotrichonymphea, including Trichonympha and Spirotrichonympha with hundreds of flagella organized in tufts or spirals).[1] Other classes include Hypotrichomonadea, Pimpavickea, Lophomonadea, Trichonymphea, Cristamonadea, Monocercomonadea, Simplicimonadea, Tritrichomonadea, and Dientamoebea, each defined by specific combinations of flagellar number, parabasal body morphology, axostyle structure, and host associations.[1]Key orders within these classes illustrate the hierarchy; for instance, Trichomonadea contains orders Trichomonadida (encompassing six families: Lacusteriidae, Trichomonadidae, and others such as those in Honigbergiellida like Hexamastigidae and Tetratrichomastigidae) and Hypotrichomonadida, while Spirotrichonymphea includes Spirotrichonymphida (with families like Spirotrichonymphidae and Holomastigotidae), alongside orders such as Cononymphida and Holomastigotoidida.[1] Additional prominent orders across classes include Cristamonadida, Devescovinida, and Trichonymphida, reflecting adaptations to symbiotic lifestyles in hosts like termites and vertebrates.[1]Classification criteria emphasize a synthesis of ultrastructural traits—such as the number and arrangement of flagella, the form of the parabasal body (e.g., fibrous or fibrillar), and karyomastigont replication patterns—with phylogenetic analyses using maximum likelihood methods on SSU rRNA and supplementary genes like actin and elongation factor 1α.[1] This approach has resolved previous non-monophyletic groupings and incorporated new taxa, such as the genusNyarlathotep in family Cthulhuidae.[1]The phylum comprises over 450 described species across more than 80 genera, though molecular surveys indicate substantial undescribed diversity, particularly in termite guts where symbiotic forms dominate.[28][1]
Diversity and Ecology
Major Lineages
The parabasalid group Trichomonadea encompasses a range of free-living and parasitic species characterized by relatively simple morphology, including up to six anterior flagella emerging from a parabasal body and a hydrogenosome for anaerobic energy production.[26] Representative parasitic forms include Trichomonas vaginalis, a common urogenital pathogen in humans transmitted sexually, and Pentatrichomonas hominis, a commensal in the large intestine of humans and other mammals that is typically non-pathogenic but can be associated with gastrointestinal disturbances in some hosts.[29][30] Another notable example is Dientamoeba fragilis, an amoeboid trichomonad lacking external flagella in its adult trophozoite stage, which inhabits the human gut and is linked to symptoms like abdominal pain and diarrhea.[31][32]The class Trichonymphea includes large, symbiotic hypermastigotes primarily found in the hindguts of wood-feeding insects. Key representatives include Trichonympha species, which dominate termite hindguts and possess up to 30 tufts of flagella arranged in spiral bands for enhanced motility and surface area in nutrient-poor environments.[33]In contrast, the Spirotrichonymphea class comprises mostly symbiotic species in the guts of arthropods, featuring highly complex, multiflagellated structures adapted for lignocellulose digestion in wood-feeding hosts.[26] Key representatives include Holomastigotoides species, symbionts in the wood-feeding cockroachCryptocercus that contribute to host cellulose breakdown through elaborate flagellar arrays, and Spirotrichonympha species in termites with spiral rows of numerous flagella.[34][35]Another major lineage, Cristamonadea, includes amoeboflagellate and trichomonad-like forms, often associated with spirochete bacteria, such as Cristamona species in termite guts that exhibit surface-attached bacterial symbionts aiding in motility and digestion.[1]Lineage-specific traits highlight evolutionary divergences within parabasalids, such as the simplified cytoskeleton in Trichomonadea—with a basic axostyle and fewer basal bodies supporting flagella—compared to the intricate, symmetrical cytoskeletal networks in Spirotrichonymphea and Trichonymphea that organize hundreds of flagella into coordinated bands.[26]Reproduction also varies, with trichomonads primarily undergoing binary fission via closed mitosis (cryptopleuromitosis) to produce daughter cells, while some hypermastigotes in lineages like Spirotrichonymphea exhibit syngamy as part of a sexual cycle, involving gamete fusion followed by meiosis in certain species.[36][37]Undescribed parabasalid diversity remains high, particularly in insect guts, where culture-independent metagenomic surveys of termite hindguts reveal hundreds of novel protist lineages unique to these hosts, suggesting extensive unrecognized forms adapted to symbiotic niches.[38][39]
Habitats and Symbiosis
Parabasalids predominantly inhabit anaerobic gut environments, where oxygen levels are low and conditions favor their hydrogenosome-based metabolism. These protists are most commonly found in the hindguts of wood-feeding invertebrates, such as termites and cockroaches, and in the rumens and intestines of ruminant vertebrates like cattle and sheep.[40][41] In termites, species such as those in the genus Trichonympha occupy the dilated hindgut, comprising a significant portion of the host's gut volume and facilitating the breakdown of lignocellulosic material in wood-based diets. Similarly, in the rumen of ruminants, parabasalids such as trichomonads contribute to the microbial community that processes fibrous plant matter, though they are less dominant compared to ciliates in these environments.[40]The symbiotic roles of parabasalids are primarily mutualistic, centered on the degradation of complex carbohydrates that hosts cannot digest independently. In termites, Trichonympha species, such as T. agilis, phagocytose wood particles and ferment cellulose into acetate, which serves as the primary energy source for the host via absorption in the hindgut; this process also produces hydrogen and carbon dioxide as byproducts, supporting the termite's ability to thrive on nutrient-poor diets. In mammalian guts, including those of ruminants, parabasalids like certain trichomonads act as commensals by hydrolyzing complex polysaccharides, thereby aiding in overall nutrient availability without direct pathogenesis in healthy hosts. These interactions enhance host fitness by improving energy extraction from recalcitrant substrates.[40]Host specificity among parabasalids is remarkably strict, with many lineages confined to particular host taxa due to co-evolutionary pressures and vertical transmission. For instance, Joenia species are associated exclusively with cockroaches, such as Periplaneta americana, where they maintain stable populations adapted to the host's hindgut pH and microbial niche. While the vast majority form obligate symbioses, rare free-living forms exist in anoxic sediments, exemplified by Pseudotrichomonas keilini, which survives in marine and freshwater mud without host dependence.[42]Ecological dynamics of parabasalid populations are influenced by host diet and environmental perturbations, leading to fluctuations that affect symbiosis stability. Dietary shifts toward higher cellulose content can increase Trichonympha densities in termite hindguts, optimizing fermentation efficiency, while low-fiber diets may reduce their abundance. Antibiotics, such as those used in veterinary medicine for ruminants, disrupt parabasalid communities by altering the gut microbiome balance, potentially decreasing protist populations and impairing carbohydrate degradation for weeks post-treatment. These dynamics underscore the sensitivity of parabasalid symbioses to host physiology and external factors.[40]
Medical and Veterinary Importance
Pathogenic Species
Trichomonas vaginalis is the primary pathogenic parabasalid affecting humans, causing trichomoniasis, a common sexually transmitted infection (STI) that leads to vaginitis in women and urethritis in men.[5] The parasite is transmitted primarily through sexual contact, with an estimated 342 million new cases globally in 2021, disproportionately affecting women of reproductive age.[43] Symptoms include vaginal discharge, itching, dysuria, and lower abdominal pain, though many infections are asymptomatic.[44]In veterinary medicine, Tritrichomonas foetus causes bovine genital trichomoniasis, a venereal disease in cattle that results in infertility, early embryonic death, and abortion.[45]Transmission occurs during breeding, with bulls acting as asymptomatic carriers harboring the parasite in the preputial cavity.[46]Pentatrichomonas hominis, another parabasalid, is associated with gastrointestinal infections in pigs, leading to diarrhea and abdominal pain, particularly in young piglets.[47] It has been detected in up to 35% of sampled pig populations in certain regions, suggesting zoonotic potential but primarily impacting swine health.[48]Pathogenesis in these species involves adhesion to host epithelial cells mediated by surface proteins such as adhesins (e.g., AP65 and AP33 in T. vaginalis), which facilitate initial attachment.[49]Cytotoxicity is driven by secreted cysteine proteases that degrade host tissues, induce inflammation, and disrupt cellular integrity, contributing to tissue damage and immune evasion.[50]Diagnosis typically relies on nucleic acid amplification tests (NAATs) for high sensitivity in detecting parasite DNA or RNA, or wet-mount microscopy to observe motile trophozoites in clinical samples.[51]Treatment for T. vaginalis infections centers on metronidazole or tinidazole, 5-nitroimidazole derivatives that disrupt parasite DNA.[44] However, metronidazole resistance has emerged, affecting 4-10% of cases globally, with 2024 proteomic studies identifying altered nitroreductase pathways in resistant strains.[52] For bovine trichomoniasis, management focuses on test-and-cull strategies due to the lack of effective vaccines or treatments, while porcine infections may respond to supportive care.[53]
Symbiotic Roles
Parabasalids play crucial symbiotic roles in the digestive systems of herbivorous insects and mammals, particularly by facilitating the breakdown of complex plant materials that hosts cannot digest independently. In the hindguts of lower termites such as Reticulitermes species and wood-feeding cockroaches like Cryptocercus punctulatus, hypermastigote parabasalids including Trichonympha and related genera dominate the protist community, comprising up to 70% of the gut volume. These flagellates harbor endosymbiotic bacteria and produce an array of glycoside hydrolases, such as endoglucanases (GH5 family), cellobiohydrolases (GH7), and β-glucosidases, which hydrolyze lignocellulose within specialized digestive vacuoles. The resulting fermentation products, primarily acetate along with H₂ and CO₂ from hydrogenosomes, provide the host with essential short-chain fatty acids for energy metabolism, while bacterial symbionts supply amino acids and vitamins, thereby enhancing overall nutrient uptake and enabling survival on a wood-based diet.[54][55]In ruminants, parabasalids such as pentatrichomonads are prevalent flagellates in the rumen and cecum of cattle, though less abundant overall than ciliates. These organisms contribute to anaerobicfermentation by digesting cellulose and other polysaccharides, producing volatile fatty acids that support host energy needs and rumen microbial balance, though their role is less dominant compared to ciliates. For instance, studies have isolated culturable pentatrichomonads from bovine rumen fluids, highlighting their adaptation to the anaerobic environment and potential involvement in fiber degradation.[41][56]The efficient lignocellulolytic capabilities of termite gut parabasalids have inspired applications in biotechnology, particularly for biofuel production. Enzymes and microbial consortia derived from these symbionts, such as Fe-hydrogenases from Pseudotrichonympha grassii, enable hydrogen and acetate generation from biomass, offering models for converting lignocellulosic waste into biofuels like ethanol and biodiesel. Research has isolated oleaginous yeasts associated with termite guts that accumulate up to 47% lipids suitable for biodiesel, while tolerating inhibitors like furfural during biomass pretreatment, demonstrating scalability for industrial biorefineries. Additionally, ongoing studies explore parabasalid-inspired probiotics to restore gut microbiota in livestock, potentially improving fermentation efficiency and animal health by mimicking natural symbiotic dynamics.[4][57][55]Disruption of parabasalid symbiosis, such as through antibiotic exposure, leads to dysbiosis and severe digestive impairments in hosts. In lower termites like Reticulitermes flavipes, treatments with antibiotics including kanamycin, metronidazole, and tetracycline drastically reduce protist densities by over 90%, collapsing lignocellulose hydrolysis and causing up to 50% declines in acetate production, which manifests as reduced survival, fecundity, and colony growth. Antibiotic-induced loss of these symbionts increases susceptibility to pathogens and prevents microbiota recovery, underscoring their indispensable role in host physiology.[58]
Evolution and Phylogeny
Evolutionary Origins
Parabasalids are thought to have originated from free-living anaerobic excavates during the early diversification of eukaryotes, representing one of the deep-branching lineages within the supergroup Excavata.[25] This ancestral state is inferred from molecular phylogenetic analyses showing parabasalids as an early-diverging group within Metamonada, with evidence of a simple, flagellated morphology adapted to anaerobic environments prior to the evolution of symbiosis.[2] A key early adaptation was the loss of aerobic mitochondrial functions, leading to the development of hydrogenosomes—anaerobic organelles that produce ATP via substrate-level phosphorylation and hydrogen gas as a byproduct, a trait shared with other ancient anaerobic lineages.[59]The development of parabasal bodies represents a defining innovation in parabasalid evolution, emerging in the ancestral excavate lineage to coordinate multiple flagella through a specialized Golgi-derived apparatus linked to basal bodies.[60] These structures facilitated enhanced motility in low-oxygen microhabitats, enabling the transition from free-living to host-associated lifestyles. Adaptation to endosymbiosis likely occurred with the radiation of arthropod hosts, with free-living forms suggesting an ancestral non-symbiotic state.[2]Fossil evidence for parabasalids is indirect but preserved in amber inclusions from termite guts, with protist nuclei exhibiting karyomastigont structures—characteristic of parabasalids—identified in Miocene (~15–20 million years ago) samples from Dominican amber, suggesting morphological stability over tens of millions of years.[61] These findings indicate ancient associations with wood-feeding insects, predating modern termite-parabasalid symbioses but aligning with the group's long-term reliance on anaerobic gut niches.Recent genomic and transcriptomic studies, including a 2024 analysis of the free-living parabasalid Pseudotrichomonas keilini, reveal patterns of gene loss consistent with early anaerobic adaptation: extensive reduction in aerobic respirationgenes (e.g., components of the electron transport chain) while retaining core anaerobic pathways such as pyruvate:ferredoxinoxidoreductase and hydrogenase for hydrogenosome function.[62] This gene content underscores the ancestral shift away from oxygen-dependent metabolism, with minimal changes in free-living relatives compared to symbiotic forms.[63]
Phylogenetic Relationships
Parabasalids, also known as Parabasalia, are positioned within the Metamonada clade of the larger Excavata supergroup, a placement consistently supported by molecular phylogenetic analyses. Multi-gene phylogenies, including those utilizing transcriptomic data from diverse parabasalid species, affirm the monophyly of Metamonada, which encompasses Parabasalia alongside Fornicata (including diplomonads such as Giardia intestinalis), oxymonads, and other lineages like Preaxostyla and Anaeramoebae. This sister relationship between Parabasalia and diplomonads within Metamonada is bolstered by 2024 phylogenomic studies employing extensive gene family datasets (e.g., 13,346 orthogroups across 43 eukaryotic genomes), which resolve deep nodes with moderate support despite challenges from anaerobic adaptations.[62][25]Within Parabasalia, recent taxonomic revisions based on integrated data divide the phylum into 11 classes, including Trichomonadea and Spirotrichonymphea, with molecular evidence from SSU rRNA genes and protein-coding markers supporting monophyly of most classes but complex inter-class relationships.[1] For example, Spirotrichonymphea groups with Cristamonadea and other classes in the Cadamassta clade, while Trichomonadea branches separately. Updated classifications confirm improved resolution across orders like Trichomonadida and Spirotrichonymphida, though some deep nodes remain challenging due to long-branch attraction in fast-evolving anaerobic lineages.[1][64]Parabasalids share certain anaerobic metabolic traits with distantly related protists like Entamoeba species in Amoebozoa, such as reliance on hydrogenosomes for ATP production and reduced mitochondrial derivatives, reflecting convergent adaptations to oxygen-poor environments; however, they differ markedly from Fornicata in cytoskeletal organization and flagellar apparatus structure. Recent genomic investigations have uncovered extensive horizontal gene transfer (HGT) from bacteria into parabasalid genomes, particularly genes involved in carbohydrate metabolism and anaerobic respiration, which has reshaped understandings of protist evolution.[62]