Campylobacter showae

Last updated

Campylobacter showae
Scientific classification OOjs UI icon edit-ltr.svg
Domain: Bacteria
Phylum: Campylobacterota
Class: "Campylobacteria"
Order: Campylobacterales
Family: Campylobacteraceae
Genus: Campylobacter
Species:
C. showae
Binomial name
Campylobacter showae
Etoh et al. 1993

Campylobacter showae is a Gram-negative, chemoheterotrophic, microaerophilic, motile bacteria belonging to the Campylobacter Genus. [1] [2] The type strain of this species, SU A4 (=ATCC 51146), was first isolated from plaque samples taken from the gingival crevices of the human oral cavity but has since also been found in colonic tissues and stool. [1] Since its discovery, C. showae has been implicated in various medical conditions including Crohn's disease, periodontitis, inflammatory bowel disease, and ulcerative colitis due to its pathogenic nature. [2] [3] [4]

Contents

Taxonomy

A pan-genome analysis, looking at a set of single-copy conserved core genes between 39 Campylobacter species, found that C. showae was most closely related to Campylobacter rectus , with C. showae and C. rectus constituting more ancient lineages within the Campylobacter genus. [5] A separate analysis of 16S rRNA sequences, comparing 1400 base positions in 21 Campylobacter, Wolinella , and Helicobacter bacterial strains, found that C. showae was most closely related to Campylobacter rectus, Campylobacter curvus , and Campylobacter concisus, with their sequences differing by 1.3%, 2.8%, and 4.9% respectively. [1] These four Campylobacter species share many similarities including their known source, being dogs and humans, and in the diseases they cause in humans, with all four being implicated in gastrointestinal diseases such as gastroenteritis or ulcerative colitis and oral diseases like periodontitis. [2]

Discovery

Campylobacter showae was first discovered in October of 1993 by researchers Yumiko Etoh, Floyd Dewhirst, Bruce Paster, Ayako Yamamoto, and Nobuichi Goto from the Showa University School of Dentistry and the Forsyth Dental Center. [1] The bacteria was obtained via isolation from human gingival crevices and was subsequently differentiated fro based on its morphological differences such as their number of flagella. [1] While the organism did show several biochemical similarities to Campylobacter curvus and Campylobacter rectus , these newly identified microbes had 2-5 flagella compared to the standard 1 in the 2 previously named organisms. [1]

Isolation and Characterization

C. showae was first isolated from plaque within the gingival crevices of 9 healthy adults using plates containing CBRCA, a medium containing reinforced clostridial agar, 5% horse blood, 0.03% China Blue, 0.2% sodium formate, 0.3% sodium fumarate and menadione. [1] Being selective for Campylobacter, Selenomonas, Fusobacterium, Veillonella, and Bacteroides strains, the resulting growth on the CBRCA plates allowed for the minimization of interference from other microorganisms and increased diagnostic sensitivity. [1] The cultures were subsequently cultivated on brain heart infusion agar plates and incubated under anaerobic conditions in a 80% nitrogen, 10% carbon dioxide, 10% hydrogen atmosphere. [1] Additionally, their ability to grow under microaerophilic conditions was tested via a Gas-Pak jar using blood agar plates with added sodium formate and fumarate at 0.2% and 0.3% concentrations respectively. [1]

Characterization of the isolated strains was performed by electron microscopy, allowing for observation of cell structure and morphology. Additionally, several tests were performed to deduce the metabolic activities of the organisms including 1) testing for the presence of catalase enzyme, characteristic of facultative anaerobes and aerobes; 2) testing for oxidase activity to detect the presence of cytochrome c oxidase, an enzyme involved in aerobic respiration; 3) detection of enzymes that allow for Indoxyl acetate hydrolysis, allowing for the differentiation of Campylobacter subspecies; 4) detection of the alkaline phosphatase and gamma-glutamyl transpeptidase enzymes, providing metabolic information for species differentiation; 5) detection of the arylsulfatase to indicate the organism's ability to hydrolyze sulfate ester bonds; 6) detection of the urease enzyme which is involved in urea hydrolysis; 7) the ability for the organism to reduce nitrate and nitrite in order to reduce hydrogen sulfide; 8) detection of lysine and ornithine decarboxylase, an enzyme that allows for the decarboxylation of lysine and ornithine amino acids; 9) resistance to antibiotics such as nalidixic acid, to differentiate species based on antibiotic resistance, and more. [1] To further differentiate species with similar phenotypes, SDS-page was used to compare the protein profiles of each bacteria, DNA-DNA homology was used to determine relatedness or genetic similarity between species, and 16S rRNA sequence comparisons were performed to deduce C. showae's phylogenetic relationships. As a result of these tests and experiments, C. showae was successfully isolated, characterized, and differentiated from its neighboring taxa. [1]

Classification as a Novel Species

C. showae was given the distinction as a new species based on phylogenetic analysis performed by 16S rRNA sequence comparisons. [1] Two previously described, but unnamed, C. showae strains were isolated and it was found that they displayed the closest relation to C. rectus, with their rRNA sequences differing at 1.35% of the positions – the same percent that Campylobacter coli and Campylobacter jejuni or Campylobacter lari and C. jejuni differed. [1] As a result of this analysis, the researchers concluded that these species should be placed in the Campylobacter genus and exhibit adequate genetic distance to justify classification as a novel species. [1]

Morphology

C. showae exhbits a curved-rod or spiral shape with round ends, measuring 2-5μm long and 0.5-0.8μm wide. [1] Although species of Campylobacter, such as C. curvus and C. rectus, primarily present only one polar flagellum, C. showae express 2-5 unipolar flagella, each with a diameter of 15-20nm. [1] Additionally, C. showae is Gram-negative and thus exhibits a cell wall composed of a cytoplasmic membrane and outer membrane in which a thin peptidoglycan layer resides in the periplasmic space. [1] C. showae cells were not observed to have any surface layer, unlike C. rectus cells which assemble an S-layer or a crystalline layer of macromolecular subunits on their cell surface for protection. [1]

Metabolism

C. showae, along with other bacteria in the Campylobacter genus, are primarily chemoheterotrophic organisms, deriving energy mainly from citric acid cycle intermediates and amino acids via oxidation. [2] Lacking the important glycolytic pathway enzyme, 6-phosphofructokinase, Campylobacter species are asaccharolytic, being unable to oxidize or ferment carbohydrates for use in its metabolism. [2] These organisms can also perform respiration to conserve energy, coupling the oxidation of formate and hydrogen to the reduction of nitrate, sulfites, fumarates, oxygen and other electron acceptors to generate ATP via electron transport phosphorylation. [2] Being a microaerophilic organism, C. showae can only tolerate an atmospheric oxygen concentration below normal atmospheric pressure, observed to grow best under atmospheric conditions of only 3%-6% Oxygen. [2]

Pathogenicity

The multiple flagella of C. showae are often found to be 2 to 3 times their attached cells in length and move in a corkscrew motion, allowing for the cell to be motile. [2] This characteristic enables C. showae and other pathogenic species of the Campylobacter genus to perform chemotaxis in response to mediating chemoattractants. [2] For instance, Campylobacter species can migrate towards metabolic substrates or electron donors and acceptors, allowing them to specifically locate and target sites for cellular invasion. [2] After overcoming the host's protective mucus layer, Campylobacter species use a multitude of adhesion-mediating factors such as CadF, an outer-membrane adhesin protein that binds to fibronectin, to facilitate adherence to host cells and induce internalization. [2] By residing within a vacuole or other membrane-bound compartment of the host cell, Campylobacter can thus avoid the defense mechanisms of the host cell and survive by utilizing its nutrients until conditions permit a cytotoxic response to be induced. [2]

Genomics

A 2019 study conducted by Tiffany Hsu et al. analyzed the genome biology and performed comparative genomics of various C. showae strains. [4] Short read Illumina sequencing was conducted for 8 strains of this bacteria with 4 being subject to PacBio RSII long read sequencing. [4] The sizes of each genome ranged from 2.1Mb-2.6Mb with an estimated >98% completeness by BUSCO with the exception of 2 strains at 92.9% and 97.6%. [4] As a result of this sequencing and subsequent functional and genomic assessments, it was found that strains of C. showae exhibit significant diversity, both genetically and phenotypically. [4] For example, the T4SS gene, which allows for the transfer of protein substrates and DNA to host cells, along with genes enabling the formation of an S-layer on the bacteria's surface, were found only in the invasive or adherent strains of C. showae. [4] On the other hand, proteins associated with CRISPR and RTX toxins (leukotoxins and hemolysins) were only found in non-invasive or non-adherent strains of C. showae. [4] Additionally, genes for methyl-accepting chemotaxis proteins were only found in C. showae strains that reside in the subgingival plaque of the oral cavity, further demonstrating the heterogeneity of C. showae strains. [4] C. showae's core genome is estimated to be comprised of around 1,284 genes, with each additional genome adding around 200 genes. [4] Within C. showae's genome, the core proteins found were DNA pol I and III, with various flagellar genes such as flhA, fliP, and fliS, also being found to be shared between strains. [4] C. showae was also found to have genes coding for the 3-subunit fumarate reductase enzyme, allowing for C. showae's fumarate metabolism, and genes coding for formate hydrogenlyase, formate tetrahydrofolate ligase, and formate dehydrogenase, allowing for C. showae's formate metabolism. [4]

Ecology

C. showae was initially isolated from human gingival crevices, residing in the human oral mucosa and within dental plaque biofilms, and has also been observed in human colonic tissues and stool. [1] [4] Exhibiting a respiratory-type metabolism, Campylobacter species require oxygen for energy production and growth. [2] However, they are microaerophilic, meaning they can only tolerate an atmospheric oxygen concentration below normal atmospheric pressure and have been observed to grow best under atmospheric conditions of only 3%-6% Oxygen. [2] C. showae, along with other human and animal pathogenic species in this genus, are mesophilic and grow best in conditions ranging from 25 to 45.4 °C (77.0 to 113.7 °F) [2] Organisms of this genus display neutrophilic characteristics, growing best in a pH range of 6.5-7.5 and are unable to survive below a pH 4.9 or above a pH of 9.0. [2] Unlike other food-borne pathogens, members of the Campylobacter genus do not display the same extent of adaptive stress responses, being vulnerable to aeration, desiccation, osmotic stress, low water activity, and high salt concentrations. [2] These characteristics are indicative of the species' adaptations to live in a host environment such as within the human oral cavity or gastrointestinal tract, in which temperature and nutrient availability remain constant. [2]

Importance

C. showae has been implicated in numerous human health conditions. For example, C. showae has historically been linked to gum disease, also known as periodontal disease. [3] Periodontal disease can be characterized by the infection of the gingiva, or gum tissue, resulting in symptoms like inflammation, bleeding on probing, and periodontal attachment loss. [3] C. showae can contribute to the development of gum disease by adhering to oral epithelial cells and releasing pro-inflammatory cytokines like IL-1β and IL-8, resulting in tissue damage. [3] [4] C. showae has also been linked to several diseases marked by persistant inflammation of a individual's digestive tract, including Crohn's disease and ulcerative colitis. [2] Having the ability to invade and colonize the intestinal mucosa, C. showae can adhere and penetrate the epithelial cells lining intestinal tract once in the gut, resulting in an immune response triggering inflammation. [2] [4] As a result of contributing and exacerbating gut inflammation, C. showae can also potentially increase the risk of developing colorectal cancer, which has been linked to chronic gut inflammation. [6] However, research must be conducted to fully understand C. showae's role in these health conditions and if a true causal relationship exists. Further research into C. showae's biology and pathogenicity can also provide valuable insights into disease prevention strategies, bacterial pathogenesis, and the consequences of bacterial dysbiosis in the human body.

Related Research Articles

<i>Escherichia coli</i> Enteric, rod-shaped, gram-negative bacterium

Escherichia coli ( ESH-ə-RIK-ee-ə KOH-ly) is a gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus Escherichia that is commonly found in the lower intestine of warm-blooded organisms. Most E. coli strains are harmless, but some serotypes such as EPEC, and ETEC are pathogenic and can cause serious food poisoning in their hosts, and are occasionally responsible for food contamination incidents that prompt product recalls. Most strains are part of the normal microbiota of the gut and are harmless or even beneficial to humans (although these strains tend to be less studied than the pathogenic ones). For example, some strains of E. coli benefit their hosts by producing vitamin K2 or by preventing the colonization of the intestine by pathogenic bacteria. These mutually beneficial relationships between E. coli and humans are a type of mutualistic biological relationship — where both the humans and the E. coli are benefitting each other. E. coli is expelled into the environment within fecal matter. The bacterium grows massively in fresh fecal matter under aerobic conditions for three days, but its numbers decline slowly afterwards.

<i>Campylobacter</i> Genus of gram-negative bacteria

Campylobacter is a type of bacteria that can cause a diarrheal disease in people. Its name means "curved bacteria", as the germ typically appears in a comma or "s" shape. According to its scientific classification, it is a genus of gram-negative bacteria that is motile.

<i>Helicobacter pylori</i> Species of bacteria

Helicobacter pylori, previously known as Campylobacter pylori, is a gram-negative, flagellated, helical bacterium. Mutants can have a rod or curved rod shape, and these are less effective. Its helical body is thought to have evolved in order to penetrate the mucous lining of the stomach, helped by its flagella, and thereby establish infection. The bacterium was first identified as the causal agent of gastric ulcers in 1983 by the Australian doctors Barry Marshall and Robin Warren.

<i>Helicobacter</i> Genus of bacteria

Helicobacter is a genus of gram-negative bacteria possessing a characteristic helical shape. They were initially considered to be members of the genus Campylobacter, but in 1989, Goodwin et al. published sufficient reasons to justify the new genus name Helicobacter. The genus Helicobacter contains about 35 species.

<i>Clostridium perfringens</i> Species of bacterium

Clostridium perfringens is a Gram-positive, bacillus (rod-shaped), anaerobic, spore-forming pathogenic bacterium of the genus Clostridium. C. perfringens is ever-present in nature and can be found as a normal component of decaying vegetation, marine sediment, the intestinal tract of humans and other vertebrates, insects, and soil. It has the shortest reported generation time of any organism at 6.3 minutes in thioglycolate medium.

<i>Campylobacter jejuni</i> Species of bacterium

Campylobacter jejuni is a species of pathogenic bacteria that is commonly associated with poultry, and is also often found in animal feces. This species of microbe is one of the most common causes of food poisoning in Europe and in the US, with the vast majority of cases occurring as isolated events rather than mass outbreaks. Active surveillance through the Foodborne Diseases Active Surveillance Network (FoodNet) indicates that about 20 cases are diagnosed each year for each 100,000 people in the US, while many more cases are undiagnosed or unreported; the CDC estimates a total of 1.5 million infections every year. The European Food Safety Authority reported 246,571 cases in 2018, and estimated approximately nine million cases of human campylobacteriosis per year in the European Union. Campylobacter jejuni infections are increasing at an alarming rate in Europe, North America, and Australia. In Africa, Asia, and the Middle East, data indicates that C. jejuni infections are endemic.

<i>Neisseria meningitidis</i> Species of bacterium that can cause meningitis

Neisseria meningitidis, often referred to as the meningococcus, is a Gram-negative bacterium that can cause meningitis and other forms of meningococcal disease such as meningococcemia, a life-threatening sepsis. The bacterium is referred to as a coccus because it is round, and more specifically a diplococcus because of its tendency to form pairs.

"Aquifex aeolicus" is a chemolithoautotrophic, Gram-negative, motile, hyperthermophilic bacterium. "A. aeolicus" is generally rod-shaped with an approximate length of 2.0-6.0μm and a diameter of 0.4-0.5μm. "A. aeolicus" is neither validly nor effectively published and, having no standing in nomenclature, should be styled in quotation marks. It is one of a handful of species in the Aquificota phylum, an unusual group of thermophilic bacteria that are thought to be some of the oldest species of bacteria, related to filamentous bacteria first observed at the turn of the century. "A. aeolicus" is also believed to be one of the earliest diverging species of thermophilic bacteria. "A. aeolicus" grows best in water between 85 °C and 95 °C, and can be found near underwater volcanoes or hot springs. It requires oxygen to survive but has been found to grow optimally under microaerophilic conditions. Due to its high stability against high temperature and lack of oxygen, "A. aeolicus" is a good candidate for biotechnological applications as it is believed to have potential to be used as hydrogenases in an attractive H2/O2 biofuel cell, replacing chemical catalysts. This can be useful for improving industrial processes.

Campylobacter upsaliensis is a gram-negative bacteria in the Campylobacter genus. C. upsaliensis is found worldwide, and is a common cause of campylobacteriosis in humans, as well as gastroenteritis in dogs and cats. Human infections are primarily associated with raw or undercooked meat and contaminated water sources, however there is some zoonotic risk associated with the spread from dogs and cats. C. upsaliensis primarily affects the gastrointestinal tract as it damages gastrointestinal epithelial cells. There are many methods for detecting C.upsaliensis including PCR and ELISA, however there is no current gold standard in detection techniques. Infection is typically self limiting, however there is antimicrobial therapy available.

Treponema denticola is a Gram-negative, obligate anaerobic, motile and highly proteolytic spirochete bacterium. It is one of four species of oral spirochetes to be reliably cultured, the others being Treponema pectinovorum, Treponema socranskii and Treponema vincentii. T. denticola dwells in a complex and diverse microbial community within the oral cavity and is highly specialized to survive in this environment. T. denticola is associated with the incidence and severity of human periodontal disease. Treponema denticola is one of three bacteria that form the Red Complex, the other two being Porphyromonas gingivalis and Tannerella forsythia. Together they form the major virulent pathogens that cause chronic periodontitis. Having elevated T. denticola levels in the mouth is considered one of the main etiological agents of periodontitis. T. denticola is related to the syphilis-causing obligate human pathogen, Treponema pallidum subsp. pallidum. It has also been isolated from women with bacterial vaginosis.

Cardiobacterium hominis /ˌkɑːrdiəʊbækˈtɪəriəm ˈhɒmɪnɪs/ is a microaerophilic, pleomorphic, fastidious, Gram-negative bacterium part of the Cardiobacteriaceae family and the HACEK group. It is most commonly found in the human microbiota, specifically the oropharyngeal region including the mouth and upper part of the respiratory tract. It is one of the causes of endocarditis, a life-threatening inflammation close to the heart's inner lining and valves. While infections caused by Cardiobacterium hominis are uncommon, various clinical manifestations are linked to the bacterium, including meningitis, septicemia, and bone infections.

Campylobacter rectus is a species of Campylobacter. It is implicated as a pathogen in chronic periodontitis, which can induce bone loss. This motile bacillus is a Gram negative, facultative anaerobe. C. rectus is associated with hypertension together with Prevotella melaninogenica and Veillonella parvula.

Campylobacter concisus is a Gram-negative, highly fastidious, mesophilic bacterium that grows under both anaerobic and microaerobic conditions with the presence of hydrogen significantly aiding growth. Motile, with either unipolar or bipolar flagella, the organisms have a characteristic spiral/corkscrew appearance and are oxidase-positive. Although the human oral cavity is the natural colonization site of the bacterium, C. concisus may also colonize the intestinal tract of some individuals. In particular, several studies have reported higher intestinal prevalence of C. concisus in patients with IBD compared to healthy controls, which has led to current speculation of the bacterium's implication in the induction of Crohn's disease.

Campylobacter mucosalis was initially isolated in 1974 by Lawson and Rowland from the lesions of porcine intestinal adenomatosis. Isolated species were gram-negative, microaerophilic and curve shaped. These organisms resembled Campylobacter sputorum in their morphological and phenotypic characteristics and were given the name Campylobacter sputorum subsp. mucosalis. A study, using DNA homology experiments, found that Campylobacter sputorum subsp. mucosalis is a distinct species and is not a subspecies of C. sputorum. Thus, its name was changed to Campylobacter mucosalis.

Lautropia mirabilis is a Gram-negative, facultatively anaerobic, oxidase- and catalase-positive, motile bacterium of the genus Lautropia and family Burkholderiaceae, isolated from the mouth of children who were infected with human immunodeficiency virus.

Treponema socranskii was isolated from gum swabs of people with periodontitis and clinically-induced periodontitis. It is a motile, helically coiled, obligate anaerobe that grows best at 37 °C, and is a novel member of its genus because of its ability to ferment molecules that other Treponema species cannot. T. socranskii’s growth is positively correlated with gingival inflammation, which indicates that it is a leading cause of gingivitis and periodontitis.

<i>Brevinema andersonii</i> Species of bacterium

Brevinema andersonii, named for John F. Anderson, who first described the organism. This organism is a Gram-negative, microaerophilic, helical shaped, chemoorganotrophic organism from the genus Brevinema. Brevinema andersonii is host associated, strains have been isolated from blood and other tissues of short-tailed shrews and white-footed mice and are infectious for laboratory mice and Syrian hamsters.B. andersonii is readily identified by restriction enzyme analysis, and SDS-PAGE, or fatty acid composition data. Another identifier for B. andersonii is the sheathed periplasmic flagella in the 1-2-1 configuration. While cells are visible by dark-field or phase-contrast microscopy, they cannot be seen when bright-field microscopy is used.

TM7x, also known as Nanosynbacter lyticus type strain TM7x HMT 952. is a phylotype of one of the most enigmatic phyla, Candidatus Saccharibacteria, formerly candidate phylum TM7. It is the only member of the candidate phylum that has been cultivated successfully from the human oral cavity, and stably maintained in vitro. and serves as a crucial paradigm. of the newly described Candidate Phyla Radiation (CPR). The cultivated oral taxon is designated as Saccharibacteria oral taxon TM7x. TM7x has a unique lifestyle in comparison to other bacteria that are associated with humans. It is an obligate epibiont parasite, or an "epiparasite", growing on the surface of its host bacterial species Actinomyces odontolyticus subspecies actinosynbacter strain XH001, which is referred to as the "basibiont". Actinomyces species are one of the early microbial colonizers in the oral cavity. Together, they exhibit parasitic epibiont symbiosis.

Thermocrinis jamiesonii is a Gram-negative bacterium that is thermophilic, growing at temperatures ranging from 70 to 85°C. It grows as a chemolithoautotroph or chemolithoheterotroph, using thiosulfate as its sole electron donor, and is obligately microaerophilic. The strain GBS1T was isolated from Great Boiling Spring, Nevada, USA.

Actinomyces massiliensis is an anaerobic, mesophilic, Gram-positive bacterium originally isolated from a human blood sample and belonging to the genus Actinomyces.

References

  1. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Etoh, Yumiko; Dewhirst, Floyd E.; Paster, Bruce J.; Yamamoto, Ayako; Goto, Nobuichi (1993). "Campylobacter showae sp. nov., Isolated from the Human Oral Cavity". International Journal of Systematic and Evolutionary Microbiology. 43 (4): 631–639. doi:10.1099/00207713-43-4-631. ISSN   1466-5034. PMID   7694633.
  2. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Silva, Willian C.; Targino, Brenda N.; Gonçalves, Amanda G.; Silva, Marcio R.; Hungaro, Humberto M. (2018), "Campylobacter : An Important Food Safety Issue", Food Safety and Preservation, Elsevier, pp. 391–430, doi:10.1016/b978-0-12-814956-0.00013-5, ISBN   978-0-12-814956-0 , retrieved 2024-04-16
  3. 1 2 3 4 Macuch, P.J.; Tanner, A.C.R. (2000). "Campylobacter Species in Health, Gingivitis, and Periodontitis". Journal of Dental Research. 79 (2): 785–792. doi:10.1177/00220345000790021301. ISSN   0022-0345. PMID   10728981.
  4. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Hsu, Tiffany; Gemmell, Matthew R.; Franzosa, Eric A.; Berry, Susan; Mukhopadhya, Indrani; Hansen, Richard; Michaud, Monia; Nielsen, Hans; Miller, William G.; Nielsen, Henrik; Bajaj-Elliott, Mona; Huttenhower, Curtis; Garrett, Wendy S.; Hold, Georgina L. (2019). "Comparative genomics and genome biology of Campylobacter showae". Emerging Microbes & Infections. 8 (1): 827–840. doi:10.1080/22221751.2019.1622455. ISSN   2222-1751. PMC   6567213 . PMID   31169073.
  5. Zhong, Chaofang; Qu, Bingpeng; Hu, Gang; Ning, Kang (2022-10-26). Garcia-Solache, Monica Adriana (ed.). "Pan-Genome Analysis of Campylobacter: Insights on the Genomic Diversity and Virulence Profile". Microbiology Spectrum. 10 (5): e0102922. doi:10.1128/spectrum.01029-22. ISSN   2165-0497. PMC   9602946 . PMID   36069574.
  6. Warren, René L; Freeman, Douglas J; Pleasance, Stephen; Watson, Peter; Moore, Richard A; Cochrane, Kyla; Allen-Vercoe, Emma; Holt, Robert A (2013). "Co-occurrence of anaerobic bacteria in colorectal carcinomas". Microbiome. 1 (1): 16. doi: 10.1186/2049-2618-1-16 . ISSN   2049-2618. PMC   3971631 . PMID   24450771.