Bacilli is a class of Gram-positive bacteria within the phylum Bacillota (formerly known as Firmicutes), characterized by low G+C content (typically less than 50%) in their DNA and predominantly rod-shaped (bacillar) morphology, although some members are spherical (cocci).[1] The name derives from the Latin word for "rod," reflecting the shape of its type genus, Bacillus.[2] This class encompasses a wide diversity of aerobic, microaerophilic, or facultatively anaerobic species, many of which are capable of forming endospores that enable survival under extreme environmental conditions such as heat, desiccation, and radiation.[3] Bacilli are ubiquitous in terrestrial and aquatic environments, including soil, water, air, and as commensals or pathogens in plants, animals, and humans.[4]Taxonomically, Bacilli is defined based on 16S rRNA gene sequence analyses and includes at least two major orders: Bacillales (the type order) and Lactobacillales.[2] The order Bacillales comprises genera such as the endospore-forming Bacillus and non-endospore-forming Listeria and Staphylococcus, with the latter including coccal forms despite the class's bacillar namesake.[5] In contrast, Lactobacillales consists mainly of non-spore-forming, acid-tolerant lactic acid bacteria (LAB) like Lactobacillus, Streptococcus, and Enterococcus, which are often associated with fermentation processes and host microbiomes.[6] Recent genomic studies have revealed further diversity within the class, including novel orders such as Erysipelotrichales and Mycoplasmatales, some of which exhibit reductive evolution leading to loss of sporulation or even peptidoglycan in their cell walls, adapting them to specialized niches like animal guts.[7] This phylogenetic grouping highlights the class's evolutionary adaptability, with members spanning free-living saprophytes to obligate parasites.Ecologically, Bacilli play crucial roles in nutrient cycling, decomposition, and symbiotic relationships. Species in Bacillales, such as Bacillus subtilis, are key decomposers in soil, breaking down organic matter and promoting plant growth through production of plant growth-promoting rhizobacteria (PGPR) compounds like auxins and siderophores.[8] LAB in Lactobacillales dominate fermented foods and the gastrointestinal microbiota of humans and animals, where they produce lactic acid to inhibit pathogens and aid digestion.[6] However, certain Bacilli are opportunistic or true pathogens; for instance, Bacillus anthracis causes anthrax, a zoonotic disease affecting livestock and humans, while Listeria monocytogenes leads to listeriosis, particularly dangerous for pregnant individuals and immunocompromised people.[3] Streptococci in Lactobacillales are responsible for infections ranging from pharyngitis to severe invasive diseases like necrotizing fasciitis.[9]In medicine and industry, Bacilli are invaluable for their biotechnological potential. Non-pathogenic strains of Bacillus species serve as probiotics to modulate gut health and enhance immune responses, while Bacillus thuringiensis produces insecticidal proteins used in biological pest control.[10] LAB are essential in food preservation, producing yogurt, cheese, and sauerkraut through fermentation, and some synthesize bacteriocins as natural preservatives.[11] Industrially, enzymes like subtilisin from B. subtilis are widely used in detergents and pharmaceuticals, underscoring the class's economic significance.[12] Despite these benefits, antibiotic resistance in pathogenic Bacilli, such as methicillin-resistant Staphylococcus aureus (MRSA), poses ongoing public health challenges.[9]
Terminology
Definition
Bacilli is a class of bacteria within the phylum Bacillota (synonym Firmicutes), encompassing primarily Gram-positive organisms characterized by their rod-shaped morphology and low mol% G+C content (typically less than 50%).[2][13] The class was formally defined by Ludwig et al. in 2010 based on phylogenetic analyses of 16S rRNA gene sequences, establishing it as a monophyletic group distinct from other Firmicutes classes like Clostridia.[13] This classification reflects the evolutionary relationships among endospore-forming and non-spore-forming lineages within the phylum.The class Bacilli includes major orders such as Bacillales, which comprises endospore-forming rods and cocci such as those in the genus Bacillus, and Lactobacillales (also known as Lactobacillus order), consisting mainly of non-spore-forming lactic acid bacteria like Lactobacillus and Streptococcus, among others.[2][13] Key diagnostic features of Bacilli include a Gram-positive cell wall structure and low mol% G+C content in their DNA, which distinguishes them from higher G+C Gram-positive phyla like Actinomycetota.The name "Bacilli" derives from the Latin word bacillus, meaning "small rod," highlighting the predominant rod-shaped (bacilliform) morphology of many members in this class, though some taxa exhibit coccal forms.[2] This etymology underscores the historical focus on cellular shape in early bacterial taxonomy, while the modern definition emphasizes molecular and phylogenetic criteria.[13]
Ambiguity with Shape and Other Taxa
The term "bacillus" (plural: bacilli) is frequently used in microbiology to describe the rod-shaped morphology of bacterial cells, regardless of their taxonomic affiliation, leading to widespread ambiguity when referring to the specific classBacilli within the phylumBacillota.[14] This morphological descriptor applies to elongated, cylindrical forms observed across diverse bacterial groups, including both Gram-positive and Gram-negative species, and predates modern phylogenetic classifications based on 16S rRNA sequencing.[14] For instance, Escherichia coli, a Gram-negative rod commonly classified as a bacillus due to its shape, belongs to the classGammaproteobacteria in the phylumPseudomonadota, not the classBacilli.[15]This confusion is compounded by the historical origins of the terminology, where "bacilli" was first employed by Ferdinand Cohn in his 1872 treatise on bacteria to denote rod-like forms, long before the establishment of formal taxonomic hierarchies in bacteriology.[16] Cohn's classification divided bacteria into four morphological groups—Sphaerobacteria (cocci), Microbacteria (short rods), Desmobacteria (filaments), and Spirobacteria (spirals)—with "bacillus" specifically introduced as a genus name for certain rod-shaped, spore-forming species like Bacillus subtilis, emphasizing form over genetic relatedness.[16] This early emphasis on morphology as a classificatory criterion has persisted in common usage, often overshadowing the class Bacilli's broader phylogenetic scope, which encompasses not only rods but also cocci such as those in the genera Staphylococcus and Streptococcus.[14]Further ambiguity arises from the overlap between the genus Bacillus—a group of primarily Gram-positive, endospore-forming rods within the order Bacillales of class Bacilli—and the class itself, leading to erroneous generalizations that equate all bacilli with members of this genus.[14] For example, Gram-negative rods like Pseudomonas species, which are also termed bacilli morphologically, are classified in the class Gammaproteobacteria of the phylum Pseudomonadota, illustrating how the term transcends phylum boundaries.[17] Such distinctions are critical in clinical and research contexts to avoid misidentification, as the class Bacilli is defined by shared genetic and physiological traits rather than shape alone.[14]
Taxonomy and Phylogeny
Historical Development
The classification of Bacilli began in the mid-19th century with Ferdinand Cohn's foundational work on bacterial taxonomy, where he included rod-shaped bacteria, including what would later be recognized as Bacilli, within the group Schizomycetes, emphasizing their fission-based reproduction and morphological diversity.[16] Cohn's 1875 publication marked an early attempt to systematize bacteria based on observable traits like shape and arrangement, laying the groundwork for subsequent refinements without yet distinguishing spore-forming capabilities.[18]By the early 20th century, classifications evolved to incorporate physiological traits, as seen in Orla-Jensen's 1909 system, which refined Cohn's framework by separating spore-forming rods (assigned to Bacillus) from non-spore-formers, highlighting endospore production as a key phenotypic marker for aerobic, Gram-positive bacteria.[19] This approach initially relied heavily on spore formation and metabolic products like lactic acid to delineate groups, with lactic acid production linking certain rods to fermentative processes, though genetic data was absent at the time. These phenotypic criteria dominated until molecular methods emerged, allowing for more precise groupings.In the late 20th century, broader taxonomic shifts occurred with Gibbons and Murray's 1978 proposal of the division Firmicutes to encompass all Gram-positive bacteria with low G+C content, initially positioning Bacilli as a class that included groups resembling Clostridia based on cell wall and spore traits.[20] A pivotal milestone came in the 1980s, when 16S rRNA sequencing enabled the separation of Bacillales (primarily aerobic spore-formers) from Lactobacillales (lactic acid producers, mostly non-spore-forming), refining the class Bacilli to reflect phylogenetic relationships over purely morphological ones.[21] This molecular approach supplanted earlier reliance on phenotypic traits, establishing Bacilli within Firmicutes as a distinct class by the 1990s. More recently, a 2021 reclassification proposed renaming Firmicutes to Bacillota for nomenclatural consistency, though it sparked debate and remains controversial among microbiologists.[22]
Current Classification
The class Bacilli belongs to the phylum Bacillota (previously designated Firmicutes), a phylum of predominantly Gram-positive bacteria characterized by low G+C content in their DNA.In contemporary taxonomy, Bacilli encompasses numerous orders (over 10 as of 2024 per LPSN), including the prominent Bacillales and Lactobacillales, reflecting distinct phylogenetic clusters based on 16S rRNA gene sequences and whole-genome analyses.[2][23]The order Bacillales encompasses families such as Bacillaceae (type family), Staphylococcaceae, Paenibacillaceae, and Listeriaceae, while Lactobacillales includes Lactobacillaceae (type family), Streptococcaceae, Leuconostocaceae, and Enterococcaceae, among others. Additional orders include Erysipelotrichales and Mycoplasmatales, highlighting recent genomic-driven expansions.[24]As documented in the List of Prokaryotic names with Standing in Nomenclature (LPSN) based on 2024 updates, the class comprises more than 10 orders, more than 50 families, and over 300 genera, with ongoing expansions driven by genomic data.[2]Prominent genera within Bacillales include Bacillus (rod-shaped endospore formers) and Staphylococcus (cocci in clusters), whereas Lactobacillales features Lactobacillus (non-spore-forming rods) and Streptococcus (cocci in chains).[5][25]Post-2010 taxonomic developments have integrated numerous novel genera from environmental isolates into Bacilli, exemplified by phylogenomic reclassifications within Bacillales, such as the elevation of environmental Bacillus-like taxa into genera like Paenibacillus and related groups.[23]
Phylogenetic Analysis
Phylogenetic analyses of the Bacilli class primarily rely on molecular methods, including 16S rRNA gene sequencing and whole-genome-based approaches. The All-Species Living Tree Project (LTP) utilizes curated 16S rRNA sequences from type strains to construct phylogenetic trees, with the most recent updates incorporating alignments up to 2020 and ongoing integrations into SILVA databases for prokaryotic taxonomy.[26] Complementing this, the Genome Taxonomy Database (GTDB) employs whole-genome sequences analyzed via concatenated alignments of 120 universal bacterial marker proteins to infer robust phylogenies, as implemented in release R10-RS226 from April 2025.[27] These methods enable rank-normalized classifications that prioritize monophyly and genome similarity metrics like average nucleotide identity (ANI).[28]Key findings from these analyses confirm that Bacilli forms a monophyletic clade within the phylum Firmicutes (now termed Bacillota in GTDB), encompassing orders such as Bacillales and Lactobacillales. Within this structure, Bacillales occupies a basal position relative to Lactobacillales in phylogenomic trees derived from marker protein concatenates, reflecting deeper divergence among aerobic, spore-forming lineages. Bacilli shares a close evolutionary relationship with the class Clostridia, another major Firmicutes group, but is distinguished by higher genomic G+C content (typically 35-55 mol%) compared to the lower values (20-45 mol%) prevalent in Clostridia, correlating with differences in metabolic versatility and environmental adaptations.[29]Discrepancies arise between GTDB and NCBI classifications, particularly in Lactobacillales, where GTDB has reclassified numerous genera based on phylogenomic coherence; for instance, the 2020 reorganization of the Lactobacillaceae family split the polyphyletic Lactobacillus genus into 25 novel genera to resolve non-monophyletic groupings unsupported by whole-genome data.[30] In contrast, NCBI retains broader, phenotype-influenced groupings, leading to inconsistencies in species assignments for over 300 lactic acid bacteria.[31]Evolutionary insights highlight the role of horizontal gene transfer (HGT) in shaping Bacilli genomes, especially for spore-forming genes; comparative analyses show that core sporulation regulators like spo0A are vertically inherited, but accessory genes involved in coat assembly exhibit HGT signatures, facilitating adaptation to diverse niches.[32] The divergence of Bacilli from other Firmicutes lineages is estimated at 2-3 billion years ago, aligning with the emergence of endospore formation near the base of the phylum during the Great Oxidation Event.[33]
Morphology and Physiology
Cellular Structure
Members of the class Bacilli are predominantly Gram-positive bacteria with diverse morphologies, typically rod-shaped (bacilliform) cells measuring 0.5–1.0 μm in width and 1.0–4.0 μm in length, though some lineages form cocci.[34][1] For instance, genera like Bacillus and Lactobacillus exhibit rod forms, while Staphylococcus consists of cocci.[1] These cells often arrange in pairs (diplobacilli) or chains (streptobacilli), depending on the species and growth conditions.[35]The cell wall structure is a defining feature for most Bacilli, consisting of a thick peptidoglycan layer that can comprise up to 90% of the wall's composition, conferring rigidity and Gram-positive staining.[36] However, certain derived orders, such as Mycoplasmatales, have undergone reductive evolution, resulting in the loss of peptidoglycan and cell walls entirely; these wall-less bacteria are adapted to specialized niches like animal hosts and cannot undergo traditional Gram staining.[7] Teichoic acids, anionic polymers of glycerol phosphate or ribitol phosphate, are covalently linked to the peptidoglycan in wall-bearing species, aiding in cation sequestration and cell wall stability.[37][38] Certain species, such as Bacillus anthracis, produce a protective capsule composed of polyglutamic acid.[3]Many rod-shaped Bacilli are motile, equipped with peritrichous flagella distributed over the cell surface to facilitate swimming in aqueous environments.[39]In the order Bacillales, a key ultrastructural adaptation is endospore formation, producing one dormant spore per cell that resists heat, desiccation, and chemicals; these spores contain dipicolinic acid, which binds calcium to stabilize the core and enhance thermal resistance.[3][40] In contrast, Lactobacillales lack this capability, as do many other orders including those with reductive evolution such as Mycoplasmatales.[41][7]
Metabolic and Growth Characteristics
Bacilli display a range of metabolic strategies reflecting the physiological diversity within the class, particularly between its two primary orders, Bacillales and Lactobacillales. Members of Bacillales are predominantly aerobic or facultatively anaerobic, relying on respiratory metabolism to generate energy, and most produce catalase to decompose hydrogen peroxide into water and oxygen, aiding in oxidative stress management.[3] In contrast, Lactobacillales are typically facultative anaerobes that favor fermentative pathways, with many species classified as lactic acid bacteria that convert sugars into lactic acid via homolactic or heterolactic fermentation, often lacking catalase activity.[42] This metabolic versatility enables Bacilli to adapt to varying oxygen availability and nutrient conditions in their environments.Growth characteristics of Bacilli are influenced by environmental factors such as temperature and pH, with optimal ranges varying by species but generally centered around mesophilic conditions. Most Bacilli thrive at temperatures between 20°C and 40°C and neutral pH values of 6 to 7, though extremes exist; for instance, psychrotolerant strains can grow below 10°C, while thermophilic members like Geobacillus species achieve optimal growth up to 70°C.[43][44] The formation of endospores in many Bacillalesspecies further enhances survival, allowing metabolic dormancy and resistance to harsh conditions like extreme heat, desiccation, or radiation until favorable growth resumes.[3]Nutritionally, Bacilli are predominantly heterotrophic, deriving energy from organic compounds such as sugars, amino acids, and proteins through chemoorganotrophic metabolism.[45] Certain genera, including Paenibacillus within Bacillales, possess the capability for biological nitrogen fixation, utilizing nitrogenase enzymes to convert atmospheric N₂ into ammonia for assimilation, which supports growth in nitrogen-limited settings.[46]Regarding antibiotic interactions, most Bacilli with intact Gram-positive cell walls exhibit sensitivity to penicillin due to the absence of an outer membrane barrier, allowing the antibiotic to target penicillin-binding proteins effectively.[47] Wall-less lineages such as those in Mycoplasmatales, however, lack peptidoglycan and are inherently resistant to β-lactam antibiotics. Resistance emerges in some wall-bearing strains through the production of β-lactamases, enzymes that hydrolyze the β-lactam ring of penicillin, with examples including class D β-lactamases identified in species like Bacillus subtilis and Bacillus pumilus.[47][48][7]
Ecology and Distribution
Habitats and Environments
Bacilli, as a class of primarily Gram-positive bacteria, exhibit a ubiquitous distribution across diverse ecosystems, including soil, water, and air. In terrestrial environments, they are particularly prevalent in soils, where genera such as Bacillus often dominate rhizospheres, the nutrient-rich zones surrounding plant roots. This prevalence stems from their ability to thrive in organic-rich substrates like decomposing plant material. Aquatic systems also harbor significant populations, with isolates recovered from both marine sediments and freshwater bodies; some form clades adapted to varied salinities.[49] Aerial dispersal further contributes to their global spread, allowing colonization of remote or transient environments.[50][51][49]Extreme environments host specialized Bacilli, demonstrating their remarkable environmental tolerance. Thermotolerant species, such as certain Bacillus strains, inhabit hot springs with temperatures exceeding 60°C, where they contribute to microbial communities shaped by high thermal gradients. In acidic soils, populations persist at pH levels as low as 4-5, with densities ranging from 5 × 10^4 to 3 × 10^6 cells per gram in regions like coastal Kerala, India. These adaptations enable survival in otherwise hostile conditions, including oligotrophic waters and desiccated lagoons. Neutral to alkaline soils support the highest abundances of Bacilli. Marine and freshwater isolates similarly show densities in the 10^5 to 10^7 CFU per liter range in sediments.[52][51][53][54]Key physiological adaptations underpin Bacilli's persistence in challenging habitats. Spore formation, a hallmark trait, confers resistance to desiccation, nutrient scarcity, and oligotrophic conditions, as seen in species like Bacillus coahuilensis from arid lagoons, where endospores endure prolonged dormancy. Biofilm production on surfaces enhances adhesion and protection in fluctuating environments, such as air-water interfaces or soil aggregates. These mechanisms allow low-level persistence in biodiversity hotspots like animal gut microbiomes, where Bacilli constitute minor components (often <1% of total bacteria),[55] and plant endospheres, where endophytic strains contribute to internal tissuediversity without dominating communities. Recent studies as of 2024 have noted shifts in Bacilli communities in response to environmental changes, such as warming in aquatic habitats.[56] Overall, these traits facilitate Bacilli's role as resilient opportunists in global microbial ecosystems.[53][57][58]
Symbiotic and Pathogenic Interactions
Bacilli exhibit diverse symbiotic relationships, particularly mutualistic associations that benefit both the bacteria and their hosts. In the human gut microbiome, species of Lactobacillus, such as L. rhamnosus and L. acidophilus, form symbiotic partnerships by fermenting carbohydrates and producing lactic acid, which aids host digestion, enhances nutrient absorption, and maintains intestinal barrier integrity against pathogens.[59][60] These bacteria also modulate the immune response, promoting tolerance and reducing inflammation, thereby supporting overall gut homeostasis.[59]In plant-associated environments, certain Bacillus species, including B. subtilis and B. velezensis, engage in mutualistic symbiosis with roots, promoting plant growth through mechanisms like phosphate solubilization. These bacteria secrete organic acids and enzymes that convert insoluble soilphosphates into bioavailable forms, enhancing phosphorus uptake and improving crop yields in nutrient-poor soils.[61][62] This interaction not only sustains bacterial populations via root exudates but also indirectly benefits the host plant by increasing resistance to stresses.[63]Many Bacilli are pathogenic, causing infections in humans and animals through opportunistic or primary mechanisms. Staphylococcus aureus, an opportunistic pathogen within the Bacilli class, commonly infects compromised skin sites such as wounds, leading to conditions like cellulitis or abscesses by colonizing and proliferating in damaged tissues.[64][65] In contrast, Streptococcus pyogenes acts as a true pathogen, primarily causing acute pharyngitis (strep throat) by invading the oropharyngeal mucosa and eliciting inflammatory responses.[66][67]Pathogenicity in Bacilli often relies on virulence factors that facilitate host invasion and immune evasion. For instance, S. pyogenes produces toxins such as streptolysins O and S, which lyse host cells, disrupt membranes, and promote tissue damage to aid bacterial spread.[68][56] Invasins like hyaluronic acid lyase enable tissue penetration, while the hyaluronic acid capsule shields bacteria from phagocytosis, allowing evasion of innate immunity.[69][70]Transmission of pathogenic Bacilli typically occurs from environmental reservoirs, such as soil or contaminated surfaces, to susceptible hosts via multiple routes. S. aureus spreads through direct contact with infected skin or fomites, as well as foodborne contamination leading to staphylococcal food poisoning.[65][64]S. pyogenes is primarily transmitted via respiratory aerosols from coughing or sneezing, facilitating person-to-person spread in close-contact settings.[56][67] These pathways underscore the importance of hygiene in preventing infections from ubiquitous Bacilli reservoirs.[71]
Significance and Applications
Pathogenic Members
Bacillus anthracis is a spore-forming bacterium responsible for anthrax, a zoonotic disease that manifests in cutaneous, inhalational, and gastrointestinal forms depending on the route of exposure.[72] Cutaneous anthrax, the most common form, results from spore entry through skin breaks, leading to localized lesions that can progress to systemic infection if untreated.[72] Inhalational anthrax occurs via aerosolized spores, causing severe respiratory distress and high mortality without prompt intervention, while gastrointestinal anthrax arises from ingesting contaminated meat, resulting in abdominal pain and bloody diarrhea.[72]Transmission is primarily spore-mediated, with spores persisting in soil and infecting humans through contact with infected animals or their products.[73] Vaccination efforts include the Sterne strain for livestock, which provides protective immunity against virulent strains, and the anthrax vaccine adsorbed (AVA) for at-risk humans, administered as a series of doses.[74][75]Staphylococcus aureus, particularly methicillin-resistant strains (MRSA), causes a spectrum of infections ranging from skin and soft tissue abscesses to severe pneumonia and sepsis.[76] MRSA infections often begin as localized skin lesions but can disseminate, leading to bloodstream infections with mortality rates exceeding 20% in hospitalized patients.[76] Many S. aureus infections are toxin-mediated, with toxic shock syndrome toxin-1 (TSST-1) implicated in toxic shock syndrome, which triggers massive cytokine release and multi-organ failure.[77] TSST-1-producing strains are associated with approximately 50% of non-menstrual toxic shock cases and nearly all menstrual cases.[77]Streptococcus pyogenes, known as group A Streptococcus (GAS), is a major cause of pyogenic infections including scarlet fever and necrotizing fasciitis, a rapidly progressive tissue-destroying condition.[78]Scarlet fever presents with fever, rash, and strawberry tongue following pharyngitis, while necrotizing fasciitis involves deep tissue necrosis and requires urgent surgical debridement.[78] Diagnosis of recent GAS infection often relies on antistreptolysin O (ASO) titers, which measure antibodies against streptolysin O and rise 1-3 weeks post-infection, aiding in confirming post-streptococcal complications like rheumatic fever.[79]Epidemiologically, anthrax reports approximately 2,000-20,000 human cases annually worldwide, predominantly in developing regions with livestock exposure.[80] As estimated in 2005, GAS diseases impose a global burden exceeding 18 million serious cases yearly, with over 500,000 deaths linked to invasive infections and sequelae like rheumatic heart disease.[81][82] S. aureus contributes significantly to antimicrobial resistance (AMR) threats, with MRSA bloodstream infections accounting for a substantial portion of the 1.27 million AMR-attributable deaths in 2019.[83] Post-2020, AMR trends in these Bacilli pathogens have intensified due to COVID-19 disruptions, including increased MRSA resistance proportions in healthcare settings and rising GAS bacteremia incidence.[84][85]
Biotechnological and Industrial Roles
Bacilli species, particularly those in the genera Bacillus and Lactobacillus, play pivotal roles in biotechnological and industrial applications due to their robust enzyme production, fermentation capabilities, and environmental remediation potential. These Gram-positive, spore-forming or lactic acid bacteria are valued for their stability under diverse conditions, enabling scalable processes in food, pharmaceutical, and environmental sectors.[86]In enzyme production, Bacillus subtilis is a primary source of subtilisin, an alkaline serine protease widely incorporated into laundry detergents for its ability to break down protein-based stains at elevated temperatures and pH levels. This enzyme's thermostability and activity in the presence of surfactants make it essential for modern detergent formulations, with industrial yields optimized through genetic engineering and fermentation techniques. Similarly, Geobacillus species, such as G. stearothermophilus, produce thermostable α-amylases used in starch hydrolysis for industries like food processing, textiles, and biofuels, where the enzymes maintain activity above 60°C, facilitating high-temperature reactions that enhance efficiency and reduce contamination risks.[50][86][87][88]Fermentation processes leverage Lactobacillus species for producing fermented dairy and vegetable products, including yogurt, cheese, and sauerkraut, where they convert sugars into lactic acid, lowering pH to preserve food and impart characteristic flavors and textures. For instance, Lactobacillus bulgaricus and Streptococcus thermophilus are key in yogurt production, initiating rapid acidification that coagulates milk proteins. Additionally, Lactobacillus acidophilus serves as a probiotic in supplements and fortified foods, promoting gut health by balancing intestinal microbiota, inhibiting pathogens, and aiding digestion, with clinical evidence supporting its role in alleviating lactose intolerance and diarrhea.[42]In bioremediation, Bacillus species demonstrate efficacy in degrading organophosphate pesticides, such as profenofos and chlorpyrifos, through enzymatic hydrolysis that mineralizes these compounds into less toxic byproducts, offering a sustainable alternative to chemical treatments in contaminated soils and waters. Bacillus subtilis strains, for example, have been shown to achieve over 80% degradation of profenofos within days under aerobic conditions. These bacteria also facilitate heavy metal sequestration via biosorption and bioaccumulation mechanisms, where extracellular polymeric substances bind ions like lead, cadmium, and chromium, reducing their bioavailability and toxicity in polluted environments; B. cereus and B. subtilis exhibit removal efficiencies up to 90% for multiple metals in lab-scale trials.[89][90][91][92]Pharmaceutical applications of Bacilli include antibiotic production, with Bacillus licheniformis serving as the main producer of bacitracin, a polypeptide antibiotic effective against Gram-positive bacteria and used topically for wound infections and in animal feed to prevent necrotic enteritis. Fermentation optimization has increased yields to over 700 IU/mL through metabolic engineering targeting precursor pathways. Furthermore, Bacillus subtilis spores function as safe, non-pathogenic vaccine vectors, displaying antigens on their surface for mucosal delivery; this platform has been explored for vaccines against rotavirus and tetanus, eliciting strong immune responses in animal models due to the spores' stability and adjuvanticity.[93][94][95]