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Bifidobacterium aerophilum sp. nov., Bifidobacterium avesanii sp. nov. and Bifidobacterium ramosum sp. nov.: Three novel taxa from the faeces of cotton-top tamarin (Saguinus oedipus L.)
Abstract
Forty-five microorganisms were isolated on bifidobacteria selective medium from one faecal sample of an adult subject of the cotton-top tamarin (Saguinus oedipus L.). All isolates were Gram-positive, catalase-negative, anaerobic, fructose-6-phosphate phosphoketolase positive, and asporogenous rod-shaped bacteria. In this study, only eight out of the forty-five strains were characterized more deeply, whereas the others are still currently under investigation. They were grouped by BOX-PCR into three clusters: Cluster I (TRE 17T, TRE 7, TRE 26, TRE 32, TRE 33, TRE I), Cluster II (TRE CT), and Cluster III (TRE MT). Comparative analysis of 16S rRNA gene sequences confirmed the results from the cluster analysis and revealed relatively low level similarities to each other (mean value 95%) and to members of the genus Bifidobacterium. All eight isolates showed the highest level of 16S rRNA gene sequence similarities with Bifidobacterium scardovii DSM 13734T (mean value 96.6%). Multilocus sequence analysis (MLSA) of five housekeeping genes (hsp60, rpoB, clpC, dnaJ and dnaG) supported their independent phylogenetic position to each other and to related species of Bifidobacterium. The G + C contents were 63.2%, 65.9% and 63.0% for Cluster I, Cluster II and Cluster III, respectively. Peptidoglycan types were A3α L-Lys–L-Thr–L-Ala, A4β L-Orn (Lys)–D-Ser–D-Glu and A3β L-Orn–L-Ser–L-Ala in Clusters I, II and III, respectively. Based on the data provided, each cluster represented a novel taxon for which the names Bifidobacterium aerophilum sp. nov. (TRE 17T =DSM 100689 = JCM 30941; TRE 26 =DSM 100690 = JCM 30942), Bifidobacterium avesanii sp. nov. (TRE CT = DSM 100685 = JCM 30943) and Bifidobacterium ramosum sp. nov. (TRE M = DSM 100688 = JCM 30944) are proposed.
... During a study exploring the presence of cultivable bifidobacterial strains in two adult subjects belonging to cotton top tamarin and emperor tamarin, we isolated and identified novel putative taxa belonging to the genus Bifidobacterium: Bifidobacterium aerophilum, Bifidobacterium avesanii, Bifidobacterium ramosum and Bifidobacterium callitrichidarum [20,26]. ...
... For discrimination of the isolates, BOX-PCR fingerprinting was carried out using the BOXA1R primer (5 -CTACGGCAAGGCGACGCTGACG-3 ) and the conditions previously described [20]. Unfortunately, all the isolates have been obtained in single strain, with the only exception for strain TRE 1 which was selected among 15 clones sharing the same BOX-PCR profiles (Data not shown). ...
... Morphological, cultural and biochemical characterization of the strains were performed at 37 • C unless otherwise stated, according to Michelini et al. [20]. ...
Four novel Gram-stain-positive, non spore forming and fructose-6-phosphate phosphoketolase-positive strains were isolated from the faeces of a cotton top tamarin (Saguinus oedipus) and an emperor tamarin (Saguinus imperator). Phylogenetic analyses based on 16S rRNA revealed that bifidobacterial strains TRE 1T exhibit close phylogenetic relatedness to Bifidobacterium catulorum DSM 103154 (96.0%) and Bifidobacterium tissieri DSM 100201 (96.0%); TRE DT and TRE HT were closely related to Bifidobacterium longum subsp. longum ATCC 15708T with similarity values of 97.4% and 97.5%, respectively; TRI 7T was closely related to Bifidobacterium tissieri DSM 100201 (96.0%). The Average Nucleotide Identity (ANI) and in silico DDH (isDDH) analysis with closest neighbour supported an independent phylogenetic position of all strains with values ranged from 74 to 85% for ANI and from 24 to 28% for isDDH. DNA base composition of the four strains was in the range of 58.3-63.5mol% G+C. Based on the phylogenetic, genotypic and phenotypic data, the strains TRE 1T, TRE DT, TRE HT and TRI 7T clearly represent four novel taxa within the genus Bifidobacterium for which the names Bifidobacterium primatium sp. nov. (type strain TRE 1T=DSM 100687T=JCM 30945T), Bifidobacterium scaligerum sp. nov. (type strain TRE DT=DSM 103140T=JCM 31792T), Bifidobacterium felsineum sp. nov. (type strain TRE HT=DSM 103139T=JCM 31789T) and Bifidobacterium simiarum sp. nov. (type strain TRI 7T=DSM 103153T=JCM 31793) are proposed.
... All strains were cultivated under anaerobic conditions and maintained in TPY broth, pH 6.9, at 37 C, unless indicated otherwise. Chromosomal DNA from all the isolates was obtained using a Wizard Genomic DNA Purification kit (Promega) according to the protocol of Michelini et al. [16]. ...
... Partial gene fragments of hsp60, rpoB, dnaG, dnaJ and clpC of strain TRI 5 T were amplified and sequenced. Sequences from the type strains of the bifidobacterial taxa were retrieved from the public database of the National Center for Biotechnology Information (NCBI) and aligned accordingly to the method of Michelini et al. [16]. ...
... Morphological, cultural and biochemical characterization of the strain TRI 5 T was performed anaerobically at 37 C, unless otherwise stated, according to standard techniques [16]. ...
Three Gram-stain-positive, non-spore-forming, microaerophilic and fructose-6-phosphate phosphoketolase positive strains were isolated from a faecal sample of an adult subject of the emperor tamarin (Saguinus imperator). Given that the isolates revealed identical BOX PCR profiles, strain TRI 5(T) was selected as a representative and characterized further. Comparative analysis of 16S rRNA gene sequence similarity revealed that strain TRI 5(T) was closely related to Bifidobacterium saguini DSM 23967(T) (96.4 %) and to Bifidobacterium longum subsp. longum ATCC 15708 (96.2 %). Multilocus sequence analyses of five housekeeping genes showed the close phylogenetic relatedness of this strain to Bifidobacterium breve DSM 20213(T) (hsp60 94.1 %), Bifidobacteriumsaguini DSM 23967(T) (clpC 91 %), Bifidobacterium avesanii DSM 100685(T) (dnaG 80.3 %), Bifidobacterium longumsubsp. infantis ATCC 15697(T) (dnaJ 85.3 %) and Bifidobacterium longumsubsp. longum ATCC 15708 (rpoB 93 %), respectively. The peptidoglycan type was A3β, with an interpeptide bridge comprising l-Orn (Lys) - l-Ser - l-Ala - l-Thr - l-Ala. The DNA G+C content of strain TRI 5(T) was 60.9 mol%. Based on the data provided, strain TRI 5(T) represents a novel species of the genus Bifidobacterium for which the name Bifidobacteriumcallitrichidarum sp. nov. is proposed. The type strain is TRI 5(T) (=DSM 103152(T)=JCM 31790(T)).
... Digestive tract of domesticated pigs represents, along with humans and primates [1][2][3], the largest reservoir of bifidobacterial diversity [4,5]. Bifidobacterium choerinum, Bifidobacterium longum subsp. ...
... Information relating to DNA G+C content, type of peptidoglycan structures and habitat were obtained from Bergey¢s Manual of Systematic Bacteriology [42] and our previous publications [14,43]. The resulting data are presented in Table 2 Nevertheless, it was recently determined in Bifidobacterium avesanii [3]. The apparent similarity between RP115 T and B. thermophilum peptidoglycan structures corresponds with relatedness found on the basis of the results of DNA-DNA and multilocus sequence analyses. ...
... They mentioned 46.5 C as the highest temperature for growth. The ability to grow at 50 C was very recently observed also in Bifidobacterium ramosum, B. avesanii and Bifidobacterium aerophilum [3]. RP115 T grows at 10 C, in contrast to the other species. ...
Fresh samples of intestinal contents of three wild pigs originating from the Central Bohemia region were examined for the presence of bifidobacterial strains. During the study, we isolated many fructose-6-phosphate phosphoketolase-positive, strictly anaerobic, irregular rod-shaped bacterial isolates. Three of them were preliminarily identified as representing a novel species of the genus Bifidobacterium because their 16S rRNA gene sequence similarity with the closest relatives of thermophilic bifidobacteria (Bifidobacterium boum DSM 20432T, Bifidobacterium thermophilum DSM 20210T, Bifidobacterium thermacidophilumsubsp. porcinum LMG 21689T, Bifidobacterium thermacidophilumsubsp. thermacidophilum DSM 15837T) was in the range of 97.9 - 98.4 %. All three bacterial isolates had identical 16S rRNA, dnaJ1, fusA, gyrB and rplB gene sequences. Isolate RP115T was chosen as a representative of the bacterial group and DNA G+C content (mol%) determination, biochemical tests and analyses of physiological and morphological characteristics, habitat and chemotaxonomic traits (peptidoglycan structure, cellular fatty acids and polar lipids profile) were performed. The DNA-DNA hybridization analyses of RP115T and species representing the group of thermophilic bifidobacteria revealed values in the range from 33 to 53 %. This fact, together with relatively low sequence similarities of particular phylogenetic markers among examined bacterial strains and the phenotyping and chemotaxonomy results obtained, indicated that the evaluated bacterial isolate should be classified as representing a separate taxon within the specific group of thermophilic bifidobacteria. The name Bifidobacterium apri (of boar) sp. nov. has been proposed for the representative strain RP115T (=CCM 8605T=DSM 100238T=LMG 28779T).
... Chromosomal DNA was extracted as previously reported [15]. The partial 16S rRNA, hsp60, clpC, dnaG, dnaJ and rpoB gene sequences of strain were amplified by PCR using the primer pairs and conditions stated in Michelini et al. [16]. ...
... Optimal growth conditions of the strain were determined in tryptone, phytone, yeast extract (TPY) broth after 48 h of Peptidoglycan type L-Orn (Lys)-L-Ser L-Glu-L-Ala-L-Lys incubation at different temperatures and at different pH in anaerobic conditions, as previously described [16]. Strain MRM 8.19 T was found to be able to survive and grow in aerobic conditions. ...
In our previous study based on hsp60 PCR-restriction fragment length polymorphism and 16S rRNA gene sequencing, we stated that the bifidobacterial strains isolated from the individual faecal samples of five baby common marmosets constituted different phylogenetically isolated groups of the genus Bifidobacterium . In that study, we also proposed that these isolated groups potentially represented novel species of the genus Bifidobacterium . Out of them, Bifidobacterium aesculapii , Bifidobacterium myosotis , Bifidobacterium tissieri and Bifidobacterium hapali , have been described recently. Another strain, designated MRM 8.19T, has been classified as member of the genus Bifidobacterium on the basis of positive results for fructose-6-phosphate phosphoketolase activity and analysis of partial 16S rRNA, hsp60, clpC, dnaJ, dnaG and rpoB gene sequences. Analysis of 16S rRNA and hsp60 gene sequences revealed that strain MRM 8.19T was related to B. tissieri DSM 100201T (95.8 %) and to Bifidobacterium bifidum ATCC 29521T (93.7 %), respectively. The DNA G+C composition was 63.7 mol% and the peptidoglycan structure was l-Orn(Lys)–l-Ser. Based on the phylogenetic, genotypic and phenotypic data reported, strain MRM 8.19T represents a novel taxon within the genus Bifidobacterium for which the name Bifidobacterium catulorum sp. nov. (type strain MRM 8.19T=DSM 103154T=JCM 31794T) is proposed.
... Bifidobacterium species are a dominant feature of the common marmoset microbiome throughout their lives. Many of the Bifidobacterium species isolated from adult animals at the UNO CRC have also been isolated from infant marmosets at other facilities (26)(27)(28)(29)(30)(31)(32)47); however, the Bifidobacterium community and its individual species have not been studied systematically and quantitatively throughout the life histories of animals at any facility. We therefore examined the Bifidobacterium community and individual species throughout the life histories of the animals using a combination of 16S rRNA amplicon sequencing and species/phylotype-specific quantitative PCR (qPCR) assays. ...
The common marmoset (Callithrix jacchus) is an omnivorous New World primate whose diet in the wild includes large amounts of fruit, seeds, flowers, and a variety of lizards and invertebrates. Marmosets also feed heavily on tree gums and exudates and they have evolved unique morphological and anatomical characteristics to facilitate gum-feeding (gummivory). In this study, we characterized the fecal microbiome of adult and infant animals from a captive population of common marmosets at the Callitrichid Research Center at the University of Nebraska at Omaha under their normal dietary and environmental conditions. The microbiome of adult animals was dominated by species of Bifidobacterium, Bacteroides, Prevotella, Phascolarctobacterium, Megamonas, and Megasphera. Culturing and genomic analysis of the Bifidobacterium populations from adult animals identified four known marmoset-associated species (B. reuteri, B. aesculapii, B. myosotis, and B. hapali) and three unclassified taxa of Bifidobacterium that are phylogenetically distinct. Species-specific qPCR confirmed that these same species of Bifidobacterium are abundant members of the microbiome throughout the life history of the animals. Genomic loci in each Bifidobacterium species encode enzymes to support growth major marmoset milk oligosaccharides during breastfeeding, however metabolic islands that can support growth on complex polysaccharide substrates in the adult captive diet (pectin, xyloglucan, and xylan) were species-specific, including loci in B. aesculapii that can support its unique ability to grow on arabinogalactan-rich tree gums.
... Members of the genus Bifidobacterium represent Gram-positive, anaerobic, non-spore forming and non-motile bacteria. Currently, the genus Bifidobacterium includes 57 taxa, representing 48 species and nine subspecies [3][4][5][6][7]. Notably, bifidobacteria have been isolated from six different ecological niches, including the gastro-intestinal tract of humans, (other) animals and insects, the oral cavity, human blood, sewage, raw milk and the water kefir fermentation process [7][8][9][10][11]. ...
A novel Bifidobacterium strain, Tam10BT, i.e. LMG 30126T, was isolated from emperor tamarin (Saguinus imperator). Cells were Gram-positive, non-motile, non-sporulating, non-haemolytic, facultative anaerobic and fructose 6-phosphate phosphoketolase-positive. Phylogenetic analyses based on 16S rRNA genes as well as multilocus sequences (representing hsp60, rpoB, dnaJ, dnaG and clpC genes) and the core genome revealed that Bifidobacterium Tam10BT exhibited close phylogenetic relatedness to Bifidobacterium tissieri DSM 100201T. Comparative analysis of 16S rRNA gene sequences confirmed the phylogenetic results showing the highest gene sequence identity with strain B. tissieri DSM 100201T (96.5 %). Furthermore, genotyping based on the genome sequence of Tam 10B, in combination with phenotypic analyses, clearly showed that strain Tam10BT is distinct from each of the type strains of the so far recognized Bifidobacterium species. The type strain Tam10BT (=LMG 30126T=CCUG 70655T) represents a novel species, for which the name Bifidobacteriumvansinderenii sp. nov is proposed.
... Lower final growth temperatures were determined in other scardovial genera. With respect to the Bifidobacteriaceae family, comparatively high final temperatures for growth were recently found in Bifidobacterium ramosum, Bifidobacterium avesanii and Bifidobacterium aerophilum [40], and much earlier in thermophilic bifidobacteria represented by Bifidobacterium thermophilum and Bifidobacterium thermacidophilum subsp. porcinum [41]. ...
Bacteria with potential probiotic applications are not yet sufficiently explored, even for animals with economic importance. Therefore, we decided to isolate and identify representatives of the family Bifidobacteriaceae, which inhabit the crop of laying hens. During the study, a fructose-6-phosphate phosphoketolase-positive strain, RP51T, with a regular/slightly irregular and sometimes an S-shaped slightly curved rod-like shape, was isolated from the crop of a 13 -month-old Hisex Brown hybrid laying hen. The best growth of the Gram-stain-positive bacterium, which was isolated using Bifidobacterium-selective mTPY agar, was found out to be under strictly anaerobic conditions, however an ability to grow under microaerophilic and aerobic conditions was also observed. Sequencing of the almost complete 16S rRNA gene (1444 bp) showed Alloscardovia omnicolens CCUG 31649T and Bombiscardovia coagulans BLAPIII/AGVT to be the most closely related species with similarities of 93.4 and 93.1 %, respectively. Lower sequence similarities were determined with other scardovial genera and other representatives of the genus Bifidobacterium. Taxonomic relationships with A. omnicolens and other members of the family Bifidobacteriaceaewere also demonstrated, based on the sequences of dnaK, fusA, hsp60 and rplB gene fragments. Low sequence similarities of phylogenetic markers to related scardovial genera and bifidobacteria along with unique features of the bacterial strain investigated within the family Bifidobacteriaceae(including the lowest DNA G+C value (44.3 mol%), a unique spectrum of cellular fatty acids and polar lipids, cellular morphology, the wide temperature range for growth (15-49 °C) and habitat) clearly indicate that strain RP51T is a representative of a novel genus within the family Bifidobacteriaceae for which the name Galliscardovia ingluviei gen. nov., sp. nov. (RP51T=DSM 100235T=LMG 28778T=CCM 8606T) is proposed.
Bifidobacteria are commensal microorganisms that typically inhabit the mammalian gut, including that of humans. As they may be vertically transmitted, they commonly colonize the human intestine from the very first day following birth and may persist until adulthood and old age, although generally at a reduced relative abundance and prevalence compared to infancy. The ability of bifidobacteria to persist in the human intestinal environment has been attributed to genes involved in adhesion to epithelial cells and the encoding of complex carbohydrate‐degrading enzymes. Recently, a putative mucin‐degrading glycosyl hydrolase belonging to the GH136 family and encoded by the perB gene has been implicated in gut persistence of certain bifidobacterial strains. In the current study, to better characterize the function of this gene, a comparative genomic analysis was performed, revealing the presence of perB homologues in just eight bifidobacterial species known to colonize the human gut, including Bifidobacterium bifidum and Bifidobacterium longum subsp. longum strains, or in non‐human primates. Mucin‐mediated growth and adhesion to human intestinal cells, in addition to a rodent model colonization assay, were performed using B. bifidum PRL2010 as a perB prototype and its isogenic perB‐insertion mutant. These results demonstrate that perB inactivation reduces the ability of B. bifidum PRL2010 to grow on and adhere to mucin, as well as to persist in the rodent gut niche. These results corroborate the notion that the perB gene is one of the genetic determinants involved in the persistence of B. bifidum PRL2010 in the human gut.
The mammalian gut is densely inhabited by microorganisms that have coevolved with their host. Amongst these latter microorganisms, bifidobacteria represent a key model to study host–microbe interaction within the mammalian gut. Remarkably, bifidobacteria naturally occur in a range of ecological niches that are either directly or indirectly connected to the animal gastrointestinal tract. They constitute one of the dominant bacterial members of the intestinal microbiota and are among the first colonizers of the mammalian gut. Notably, the presence of bifidobacteria in the gut has been associated with several health-promoting activities. In this review, we aim to provide an overview of current knowledge on the genetic diversity and ecology of bifidobacteria. Furthermore, we will discuss how this important group of gut bacteria is able to colonize and survive in the mammalian gut, so as to facilitate host interactions.
Five Bifidobacterium strains, VB23 T , VB24 T , VB25 T , VB26 T and VB31 T , were isolated from chimpanzee ( Pan troglodytes ), cotton-top tamarin ( Saguinus oedipus ), Goeldi’s marmoset ( Callimico goeldii ), moustached tamarin ( Saguinus mystax ) and patas monkey ( Erythrocebus patas ), respectively, which were kept in two Czech zoos. These strains were isolated from faecal samples and were Gram-positive, non-motile, non-sporulating, anaerobic and fructose-6-phosphate phosphoketolase-positive. Phylogenetic analyses based on 16S rRNA revealed close relatedness between VB23 T and Bifidobacterium angulatum LMG 11039 T (96.0 %), VB24 T and Bifidobacterium pullorum subsp. pullorum DSM 20433 T (96.1 %), VB25 T and Bifidobacterium goeldii LMG 30939 T (96.5 %), VB26 T and Bifidobacterium imperatoris LMG 30297 T (98.1 %), and VB31 T and B . angulatum LMG 11039 T (99.40 %). Internal transcribed spacer profiling revealed that VB23 T , VB24 T , VB25 T , VB26 T and VB31 T had highest similarity to Bifidobacterium breve LMG 13208 T (77.2 %), Bifidobacterium longum subsp. infantis ATCC 15697 T (85.8 %), Bifidobacterium biavatii DSM 23969 T (76.9 %), B. breve LMG 13208 T (81.2 %) and B. angulatum LMG 11039 T (88.2 %), respectively. Average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) analyses with their closest neighbours supported the independent phylogenetic positions of the strains with values between 86.3 and 94.3 % for ANI and 25.8 and 54.9 % for dDDH. These genomic and phylogenetic analyses suggested that the evaluated strains were novel Bifidobacterium species named Bifidobacterium erythrocebi sp. nov. (VB31 T =DSM 109960 T =CCUG 73843 T ), Bifidobacterium moraviense sp. nov. (VB25 T =DSM 109958 T =CCUG 73842 T ), Bifidobacterium oedipodis sp. nov. (VB24 T =DSM 109957 T =CCUG 73932 T ), Bifidobacterium olomucense sp. nov. (VB26 T =DSM 109959 T =CCUG 73845 T ) and Bifidobacterium panos sp. nov. (VB23 T =DSM 109963 T =CCUG 73840 T ).
A Gram-stain-positive, facultative anaerobic, rod-shaped bacteria isolated from the small intestine of a mini pig was designated as strain YH-lac9 T . 16S rRNA gene sequence analysis revealed that the strain belongs to the genus Lentilactobacillus and is closely related to Lentilactobacillus senioris JCM 17472 T , Lentilactobacillus rapi JCM 15042 T and Lentilactobacillus diolivorans JCM 13927 T , with 97.6, 96.2 and 95.7 % sequence similarity, respectively. Analysis of housekeeping gene sequences ( pheS and recA ) revealed that the strain formed a sub-cluster with L. senioris , supporting the results of 16S rRNA gene sequences analysis. The average nucleotide identity value for YH-lac9 T and the most closely related strain is 74.1 %. The main fatty acids are C 18 : 1 ω9 c , summed feature 7, C 16 : 0 and summed feature 8. The G+C content of the genomic DNA is 37.8 mol%. In view of its chemotaxonomic, phenotypic and phylogenetic properties, YH-lac9 T (=KCTC 25005=JCM 33997) represents a novel taxon. The name Lentilactobacillus kribbianus sp. nov. is proposed.
Seven bifidobacterial strains were isolated from the faeces of two adult males of the two-toed sloth (Choloepus didactylus) housed in Parco Natura Viva, in Italy. Comparative sequence analysis of 16S rRNA and of five housekeeping (hsp60, rpoB, clpC, dnaJ, dnaG) genes revealed that these strains were classified into two clusters. On the basis of 16S rRNA gene sequence similarity, the type strain of Bifidobacterium catenulatum subsp. kashiwanohense DSM 21854T (95.4 %) was the closest neighbour to strain in Cluster I (BRDM 6T), whereas the type strain of Bifidobacterium dentium DSM 20436T (values were in the range of 98‒99.8 %) was the closest neighbour to the other six strains in Cluster II. The average nucleotide identity (ANI) values of BRDM 6T and of strains in Cluster II with the closely related type strains were 76.0 and 98.9 % (mean value) respectively. Therefore, genotyping based on the genome sequence of the strain BRDM 6T combined with phenotypic analyses clearly revealed that the strain BRDM 6T represents a novel species for which the names Bifidobacterium choloepi sp. nov. (BRDM 6T = NBRC 114053T = BCRC 81222T) is proposed.
Exorista larvarum (L.) (Diptera: Tachinidae), a larval parasitoid of Lepidoptera, can be reared from egg to fecund adult on artificial media composed of crude components. The standard in vitro culture is performed in 24‐well plastic rearing plates. Exorista larvarum eggs, removed from superparasitized larvae of Galleria mellonella (L.) (Lepidoptera: Pyralidae), are individually placed in the wells, each containing a cotton swab soaked in liquid medium. The plates are then sealed until parasitoid puparium formation. To avoid contamination by microorganisms, the artificial medium is routinely supplemented with 0.01% solution of gentamicin. Experiments were carried out to assess whether this broad‐spectrum antibiotic may be replaced with hydrolate of Monarda fistulosa L. (Lamiaceae), which was selected due to its high in vitro activity against pathogenic microorganisms for humans and plants. The hydrolate was either supplemented to the artificial medium (0.5% wt/wt) (first experiment) or placed in an empty well (200 μl) of the rearing plate, to be supplied as saturated air due to evaporation (second experiment). In both experiments, a standard medium with gentamicin and an antimicrobial‐free medium were maintained as positive and negative controls, respectively. In the first experiment, in the hydrolate‐supplemented medium fewer E. larvarum completed egg‐to‐adult development than in the standard medium, but significantly more parasitoid developed from egg to adult compared to the antimicrobial‐free medium. No significant difference was found between the numbers of eggs laid by the females obtained from the standard medium vs. those from the hydrolate‐supplemented medium. In the second experiment, the hydrolate‐saturated air significantly decreased E. larvarum egg hatching, puparium formation, and female fecundity compared to the standard medium. In perspective, M. fistulosa hydrolate supplemented to the artificial media for E. larvarum may be considered as a promising candidate to replace the gentamicin solution, as suggested also by the microbiological analyses of the media, performed at various growth stages of the parasitoid in a separate trial. Conversely, the hydrolate‐saturated air treatment was deemed unsuitable.
Three bifidobacterial Gram-stain-positive, non-spore forming and fructose-6-phosphate phosphoketolase-positive strains, SMA1T, SMB2 and SMA15T were isolated from the faeces of two adult males of the squirrel monkey (Saimiri sciureus).
On the basis of 16S rRNA gene sequence similarities, the type strain of Bifidobacterium primatium DSM 100687T (99.3%; similarity) was the closest neighbour to strains SMA1T and SMB2, whereas the type strain of Bifidobacterium stellenboschense DSM 23968T (96.5%) was the closest neighbour to strain SMA15T. The average nucleotide identity (ANI) values of SMA1T and SAM15T with the closely related type strains were 93.7% and 88.1%, respectively. The in silico DNA‒DNA hybridization values with the closest neighbours were 53.1% and 36.9 %, respectively. GC contents of strains SMA1T and SMA15T were 63.6 and 66.4 mol%, respectively. Based on the phylogenetic, genotypic and phenotypic data obtained, the strains SMA1T and SMA15T clearly represent two novel taxa within the genus Bifidobacterium for which the names Bifidobacterium saimiriisciurei sp. nov. (type strain SMA1T = BCRC 81223T = NBRC 114049T= DSM 106020T) and Bifidobacterium platyrrhinorum sp. nov. (type strain SMA15T = BCRC 81224T = NBRC 114051T= DSM 106029T) are proposed.
In recent years, bifidobacterial populations in the gut of various monkey species have been assessed in several ecological surveys, unveiling a diverse, yet unexplored ecosystem harboring novel species. In the current study, we investigated the species distribution of bifidobacteria present in 23 different species of primates, including human samples, by means of 16S rRNA microbial profiling and ITS bifidobacterial profiling. Based on the observed bifidobacterial‐host co‐phylogeny, we found a statistically significant correlation between the Hominidae family and particular bifidobacterial species isolated from humans, indicating phylosymbiosis between these lineages. Furthermore, phylogenetic and glycobiome analyses, based on 40 bifidobacterial species isolated from primates, revealed that members of the Bifidobacterium tissieri phylogenetic group, which are typical gut inhabitants of members of the Cebidae family, descend from an ancient ancestor with respect to other bifidobacterial taxa isolated from primates. This article is protected by copyright. All rights reserved.
Two Bifidobacterium strains, i.e., 2176BT and 2177BT, were isolated from Golden-Headed Lion Tamarin (Leontopithecus chrysomelas) and Goeldi's monkey (Callimico goeldii). Isolates were shown to be Gram-positive, non-motile, non-sporulating, facultative anaerobic and d-fructose 6-phosphate phosphoketolase-positive. Phylogenetic analyses based on 16S rRNA sequences, multilocus sequences (including hsp60, rpoB, dnaJ, dnaG and clpC genes) and the core genome revealed that bifidobacterial strains 2176BT and 2177BT exhibit close phylogenetic relatedness to Bifidobacterium felsineum DSM 103139T and Bifidobacterium bifidum LMG 11041T, respectively. Further genotyping based on the genome sequence of the isolated strains combined with phenotypic analyses, clearly show that these strains are distinct from each of the type strains of the so far recognized Bifidobacterium species. Thus, Bifidobacterium cebidarum sp. nov. (2176BT=LMG 31469T=CCUG 73785T) and Bifidobacterium leontopitheci sp. nov. (2177BT=LMG 31471T=CCUG 73786T are proposed as novel Bifidobacterium species.
Fifteen bifidobacterial strains were obtained from faeces of Rousettus aegyptiacus; after grouping them by RAPD PCR only eight were selected and characterized. Analysis of 16S rRNA and of five housekeeping (hsp60, rpoB, clpC, dnaJ, dna G) genes revealed that these eight strains were classified into five clusters: Cluster I (RST 8 and RST 16T), Cluster II (RST 9T and RST 27), Cluster III (RST 7 and RST 11), Cluster IV (RST 19), Cluster V (RST 17) were closest to Bifidobacterium avesanii DSM 100685T (96.3%), Bifidobacterium callitrichos DSM 23973T (99.2% and 99.7%), Bifidobacterium tissieri DSM 100201T (99.7 and 99.2%), Bifidobacterium reuteri DSM 23975 T (98.9%) and Bifidobacterium myosotis DSM 100196T (99.3%), respectively. Strains in Cluster I and strain RST 9 in Cluster II could not be placed within any recognized species while the other ones were identified as known species. The average nucleotide identity values between two novel strains, RST 16T and RST 9T and their closest relatives were lower than 79% and 89%, respectively. In silico DNA-DNA hybridization values for those closest relatives were 32.5 and 42.1%, respectively. Phenotypic and genotypic tests demonstrated that strains in Cluster I and RST 9T in Cluster II represent two novel species for which the names Bifidobacterium vespertilionis sp. nov. (RST 16T=BCRC 81138T=NBRC 113380T=DSM 106025T ; RST 8=BCRC 81135=NBRC 113377) and Bifidobacterium rousetti sp. nov. (RST 9T=BCRC 81136T=NBRC 113378T=DSM 106027T) are proposed.
A novel irregularly shaped and slightly curved rod bacterial strain, GLDI4/2T, showing activity of fructose 6-phosphate phosphoketolase was isolated from a faecal sample of an adult gelada baboon (Theropithecus gelada). Phylogenetic analyses based on 16S rRNA genes as well as multilocus sequences (representing fusA, gyrB and xfp genes) and the core genome revealed that GLDI4/2T exhibited phylogenetic relatedness to Alloscardovia omnicolens DSM 21503T and to Alloscardovia macacae DSM 24762T. Comparative analysis of 16S rRNA gene sequences confirmed the phylogenetic results showing the highest gene sequence identity with strain A. omnicolens DSM 21503T (96.0 %). Activities of α- and β-gluco(galacto)sidases were detected in strain GLDI4/2T, which is characteristic for almost all members of the family Bifidobacteriaceae. Compared to other Alloscardovia species its DNA G+C content (43.8 mol%) was very low. Phylogenetic studies and the evaluation of phenotypic characteristics, including the results of biochemical, physiological and chemotaxonomic analyses, confirmed the novel species status for strain GLDI4/2T, for which the name Alloscardoviatheropitheci sp. nov. is proposed. The type strain is GLDI4/2T (=DSM 106019T=JCM 32430T).
Aim:
to characterise four novel autochthonous bifidobacteria isolated from monkey faeces and a B. lactis strain isolated from chicken faeces by evaluating their technological and biological/functional potential to be used as probiotics. Different stressors, including food process parameters and storage, can affect their viability and functionality.
Methods and results:
The resistance to frozen-storage, tolerance to lyophilisation and viability during storage, thermal, acidic and simulated gastric resistance, surface hydrophobicity and antimicrobial activity against pathogens were studied. Bifidobacterium lactis Bb12 and INL1 were used as reference strains. The results obtained demonstrated that the new isolates presented strain-dependent behaviour. Good results were obtained for thermal resistance, frozen-storage at -80°C and lyophilised powders maintained at 5°C. Cell viability during refrigerated storage was higher when the strains were resuspended in milk at pH 5.0 than at 4.5. The surface hydrophobicity ranged between 7 and 98% depending on the strain. The simulated gastric resistance was improved for the strains incorporated in cheese. Regarding antimicrobial activity, bifidobacteria isolated from monkey presented higher inhibitory capacity than the reference strains.
Conclusion:
this research provides a deeper insight into new strains of bifidobacteria isolated from primates and chicken that have not been previously characterised for their potential use in dairy products and confirm the most robust stress tolerance of B. lactis.
Significance and impact of the study:
the possibility of expanding the available bifidobacteria with the potential to be added to a probiotic food necessarily implies characterising them from different points of view, especially when considering unknown species. For monkey-isolates (which showed higher antimicrobial activity against pathogens), more in-depth knowledge is needed before applying strategies to improve their performance. On the contrary, the chicken-isolate B. lactis P32/1 showed similar behaviour to the references B. lactis strains; therefore, it could be considered as a potential probiotic candidate. This article is protected by copyright. All rights reserved.
The genus Bifidobacterium is purported to have beneficial consequences for human health and is a major component of many gastrointestinal probiotics. Although species of Bifidobacterium are generally at low relative frequency in the adult human gastrointestinal tract, they can constitute high proportions of the gastrointestinal communities of adult marmosets. To identify genes that might be important for the maintenance of Bifidobacterium in adult marmosets, ten strains of Bifidobacterium were isolated from the feces of seven adult marmosets, and their genomes were sequenced. There were six B. reuteri strains, two B. callitrichos strains, one B. myosotis sp. nov. and one B. tissieri sp. nov. among our isolates. Phylogenetic analysis showed that three of the four species we isolated were most closely related to B. bifidum, B. breve and B. longum, which are species found in high abundance in human infants. There were 1357 genes that were shared by at least one strain of B. reuteri, B. callitrichos, B. breve, and B. longum, and 987 genes that were found in all strains of the four species. There were 106 genes found in B. reuteri and B. callitrichos but not in human bifidobacteria, and several of these genes were involved in nutrient uptake. These pathways for nutrient uptake appeared to be specific to Bifidobacterium from New World monkeys. Additionally, the distribution of Bifidobacterium in fecal samples from captive adult marmosets constituted as much as 80% of the gut microbiome, although this was variable between individuals and colonies. We suggest that nutrient transporters may be important for the maintenance of Bifidobacterium during adulthood in marmosets.
HIGHLIGHTS
• Bifidobacterium from marmosets are close relatives of those from tamarins.
• 72% of adult marmosets had relative abundances of Bifidobacterium greater than 10%.
• Genes involved in nutrient uptake may allow Bifidobacterium to flourish in adult marmosets.
Five Bifidobacterium strains, i.e. 2020BT, 2028BT, 2033BT, 2034BT and 2036BT, were isolated from European beaver (Castor fiber), Goeldi's marmoset (Callimicogoeldii), black-capped squirrel monkey (Saimiriboliviensissubsp. peruviensis) and Patagonian mara (Dolichotispatagonum). All of these isolates were shown to be Gram-positive, facultative anaerobic, d-fructose 6-phosphate phosphoketolase-positive, non-motile and non-sporulating. Phylogenetic analyses based on 16S rRNA gene sequences, multilocus sequences (including hsp60, rpoB, dnaJ, dnaG and clpC genes) and the core genome revealed that bifidobacterial strains 2020BT, 2028BT, 2033BT, 2034BT and 2036BT exhibit close phylogenetic relatedness to Bifidobacteriumbiavatii DSM 23969T, Bifidobacteriumbifidum LMG 11041T, Bifidobacteriumchoerinum LMG 10510T, Bifidobacteriumgallicum LMG 11596T, Bifidobacteriumimperatoris LMG 30297T, Bifidobacteriumitalicum LMG 30187T and Bifidobacteriumvansinderenii LMG 30126T, respectively. Further genotyping based on the genome sequence of the isolated strains combined with phenotypic analyses, clearly show that these strains are distinct from each of the type strains of the so far recognized Bifidobacterium species. Thus, Bifidobacteriumcastoris sp. nov. (2020BT=LMG 30937T=CCUG 72816T), Bifidobacteriumcallimiconis sp. nov. (2028BT=LMG 30938T=CCUG 72814T), Bifidobacteriumsamirii sp. nov. (2033BT=LMG 30940T=CCUG 72817T), Bifidobacteriumgoeldii sp. nov. (2034BT=LMG 30939T=CCUG 72815T) and Bifidobacteriumdolichotidis sp. nov. (2036BT=LMG 30941T=CCUG 72818T) are proposed as novel Bifidobacterium species.
Bifidobacterium is a diverse genus of anaerobic, saccharolytic bacteria that colonize many animals, notably humans and other mammals. The presence of these bacteria in the gastrointestinal tract represents a potential coevolution between the gut microbiome and its mammalian host mediated by diet. To study the relationship between bifidobacterial gut symbionts and host nutrition, we analyzed the genome of two bifidobacteria strains isolated from the feces of a common marmoset (Callithrix jacchus), a primate species studied for its ability to subsist on host-indigestible carbohydrates. Whole genome sequencing identified these isolates as unique strains of Bifidobacterium callitrichos. All three strains, including these isolates and the previously described type strain, contain genes that may enable utilization of marmoset dietary substrates. These include genes predicted to contribute to galactose, arabinose, and trehalose metabolic pathways. In addition, significant genomic differences between strains suggest that bifidobacteria possess distinct roles in carbohydrate metabolism within the same host. Thus, bifidobacteria utilize dietary components specific to their host, both humans and non-human primates alike. Comparative genomics suggests conservation of possible coevolutionary relationships within the primate clade.
Six Bifidobacterium strains, i.e., Goo31D, Ham19E, Rab10A, Tam1G, Uis4E and Uis1B, were isolated from domestic goose (Anser domesticus), European hamster (Cricetus cricetus), European rabbit (Oryctolagus cuniculus), emperor tamarin (Saguinus imperator) and pygmy marmoset (Callithrix pygmaea). Cells are Gram-positive, non-motile, non-sporulating, facultative anaerobic and fructose 6-phosphate phosphoketolase-positive. Phylogenetic analyses based on 16S rRNA, ITS-, multilocus- sequences and the core genome revealed that bifidobacterial strains Goo31D, Ham19E, Rab10A, Tam1G, Uis4E and Uis1B exhibit close phylogenetic relatedness with Bifidobacterium choerinum LMG 10510, Bifidobacterium hapali DSM 100202, Bifidobacterium saguini DSM 23967 and Bifidobacterium stellenboschense DSM 23968. Genotyping based on the genome sequence of the isolated strains combined with phenotypic analyses, clearly show that these strains are distinct from each of the type strains of the so far recognized Bifidobacterium species. Thus, Bifidobacterium anseris sp. nov. (Goo31D=LMG 30189T =CCUG 70960T), Bifidobacterium criceti sp. nov. (Ham19E=LMG 30188T =CCUG 70962T), Bifidobacterium imperatoris sp. nov. (Tam1G=LMG 30297T =CCUG 70961T), Bifidobacterium italicum sp. nov. (Rab10A=LMG 30187T =CCUG 70963T), Bifidobacterium margollesii sp. nov. (Uis1B=LMG 30296T =CCUG 70959T) and Bifidobacterium parmae sp. nov. (Uis4E=LMG 30295T =CCUG 70964T) are proposed as novel Bifidobacterium species.
Bifidobacterium sp. strains TRE 1, TRE D, TRE H, and TRI 7 were isolated from two tamarins housed in Parco Natura Viva, Garda Zoological Park S.r.l. (Bussolengo, Verona, Italy). These strains belong to four putative novel species of the genus Bifidobacterium . The genome sizes were 2.7 Mb for TRE 1, 2.7 Mb for TRE D, 2.4 Mb for TRE H, and 2.7 Mb for TRI 7. The average GC contents were 63.18% for TRE 1, 58.27% for TRE D, 57.11% for TRE H, and 63.79% for TRI 7.
Nonhuman primates represent a major component of the frugivore biomass in several rain-forest communities. Although there is considerable evidence that prosimians, monkeys, and apes serve as dispersal agents for many tropical trees, little attention has been paid to the more basic questions of why certain species of primates swallow and void seeds, and what, if any, are the advantages to an animal of having a large, hard, bolus pass through its digestive tract. We examine patterns of fruit-eating and seed-swallowing in two species of free-ranging tamarins: Saguinus mystax and Saguinus geoffroyi. Fruits commonly eaten by tamarins contain large seeds surrounded by a fibrous and adhesive pulp or arilate seed coat. They generally swallow seeds and pulp together. Intact seeds are voided over a 1- to 3-h period. Measurements of 132 seeds naturally voided by Panamanian tamarins average 11.2 mm in length and 0.3 g. The greatest number of large seeds contained in the digestive tract of a single animal at one time was 13. In the case of moustached tamarins, we collected 220 seeds. Average seed length is 11.9 mm and average seed weight is 0.3 g. At the time of capture, one animal had 26 seeds in its digestive tract. In both tamarin species, there is evidence of sex-based differences in feeding behavior. Adult female moustached and Panamanian tamarins swallowed and voided seeds of larger size than adult males did. Seed size is positively correlated with pulp weight (p
The occurrence and species distribution of bifidobacteria in the digestive tract of important representatives of social insects such as ants, bees, wasps and bumblebees as well as the incidence of bifidobacteria in fecal samples of several species of vertebrates represented mainly by reptiles was assigned by culture-independent method based on DGGE and real time PCR. Bifidobacteria were present in the gut of most social insects--honey bees, wasps, cockroaches and bumblebees, except for ants. In honey bees, where the counts of bifidobacteria ranged from 2 to 8% of the total bacteria, the most common species seemed to be Bifidobacterium indicum. Proportion of bifidobacteria was found in broad range from 0.1 to 35-37% in wasps and cockroaches; the variance of bifidobacteria in bumblebees was lower, ranging from 1 to 7% of total bacterial count. Among studied vertebrates, the detectable presence of bifidobacteria was found only in trout (1.1%) and geckos (0.2%), but large amount of these bacteria was observed in Vietnamese box turtle, where bifidobacteria represented nearly one-fourth (22%) of total bacterial counts.
Forty strains of bifidobacteria were isolated from the faeces of two adult subjects of black lemur, Eulemur macaco. Twenty-five were identified as Bifidobacterium lemurum, the novel species recently described in Lemur catta. All other isolates resulted Gram-positive-staining, non-spore-forming, fructose-6-phosphate phosphoketolase positive, microaerophilic, irregular rod-shaped bacteria that often resembled Y or V shapes cells. Typing techniques revealed these isolates were nearly identical and strain LMM_E3T was chosen as representative and characterized further. Phylogenetic analysis based on 16S rRNA gene sequences clustered this isolate inside the genus Bifidobacterium and showed the highest levels of sequence similarities with Bifidobacterium lemurum DSM 28807T (99.6%), with Bifidobacterium pullorum LMG 21816T and Bifidobacterium longum subsp. infantis ATCC 15697T (96.4 % and 96.3%, respectively). Analysis of hsp60 gene sequences revealed that strain LMM_E3T was also closely related to Bifidobacterium stellenboschense DSM 23968T (93.3%). DNA-DNA reassociation value with the closest neighbour B. lemurum DSM 28807T was found to be 65.4%. The DNA base composition was 62.3 mol% G+C. Strain LMM_E3T showed a peptidoglycan structure which has not been detected in bifidobacteria so far: A3α L-Lys - L-Ser - L-Thr - L-Ala. Based on the phylogenetic, genotypic and phenotypic data, strain LMM_E3 T represents a novel species within the genus Bifidobacterium for which the name Bifidobacterium eulemuris sp. nov. is now proposed; the type strain is LMM_E3T (=DSM 100216T; = JCM 30801T).
In a previous study on bifidobacterial distribution in New World Monkeys, six Bifidobacteriaceae strains were isolated from faecal samples of baby common marmosets (Callitrix jacchus L.). All the isolates were gram-positive-staining, anaerobic, asporogenous and fructose-6-phosphate phosphoketolase positive. Comparative analysis of 16S rRNA gene sequences revealed relatively low levels of similarity (maximum identity 96%) to members of the genus Bifidobacterium, and placed the isolates in three independent clusters: Cluster I (MRM_5.9T and MRM_5.10) and Cluster III (MRM_5.18T and MRM_9.02) respectively showed 96.4% and 96.7% homology with Bifidobacterium callithricos DSM 23973T, while Cluster II (MRM_8.14T and MRM_9.14) had 95.4% with Bifidobacterium stellenboschense DSM 23968T. Phylogenetic analysis of hsp60 and clpC partial gene sequences supported an independent phylogenetic position, to each other and to the related species B. callithricos DSM 23973T and B. stellenboschense DSM 23968T. Clusters I, II and III respectively show a GC content of 64.9‒65.1%, 56.4‒56.7% and 63.1‒63.7%. The major cellular fatty acid for MRM_5.9T was C14:0, C16:0 and C18:1 ω9c DMA, while C16:0 was markedly present in strains MRM_5.18T and MRM_8.14T, followed by C18:1 ω9c and C14:0. Biochemical profiles and growth parameters were recorded for all the isolates. Based on the data provided, the clusters represent three novel species for which the names Bifidobacterium myosotis sp. nov. (type strain: MRM_5.9T =DSM 100196T = JCM 30796T), Bifidobacterium hapali sp. nov. (type strain: MRM_8.14T = DSM 100202T = JCM 30799T) and Bifidobacterium tissieri sp. nov. (type strain: MRM_5.18T = DSM 100201T = JCM 30798T) are proposed.
Bifidobacteria were isolated from the gut of Bombus lapidarius, Bombus terrestris and Bombus hypnorum bumble bees by direct isolation on modified trypticase phytone yeast extract agar. The MALDI-TOF MS profiles of four isolates (LMG 28292(T), R-53560, R-53124, LMG 28626) were found to be identical and did not cluster with the profiles of established Bifidobacterium species. Analysis of the 16S rRNA gene sequence of strain LMG 28292(T) revealed that LMG 28292(T) is most closely related to the Bifidobacterium bohemicum type strain (96.8 %), which was also isolated from bumble bee gut specimens. The hsp60 gene of strain LMG 28292(T) shows 85.8 % sequence similarity to that of the B. bohemicum type strain. The (GTG)5-PCR profiles and the hsp60 sequences of all four isolates were indistinguishable; however, three different phenotypes were observed among the four isolates by means of the API 50CHL microtest system. Based on the phylogenetic, genotypic and phenotypic data, we propose to classify the four isolates within the novel species Bifidobacterium commune sp. nov., with LMG 28292(T) (= DSM 28792(T)) as the type strain.
Despite the generally accepted importance of bifidobacteria as probiotic components of the human intestinal microflora and their use in health promoting foods, there is only limited information about their phylogenetic position, physiology and underlying genetics. In the last few years numerous molecular approaches have emerged for the identification and characterization of bifidobacterial strains. Their use, in conjunction with traditional culturing methods, has led to a polyphasic taxonomy which has significantly enhanced our knowledge of the role played by these bacteria in the human intestinal ecosystem. The recent adaptation of culture-independent molecular tools to the fingerprinting of intestinal and food communities offers an exciting opportunity for revealing a more detailed picture of the true complexity of these environments. Furthermore, the availability of bifidobacterial genome sequences has advanced knowledge on the genetics of bifidobacteria and the effects of their metabolic activities on the intestinal ecosystem. The release of a complete Bifidobacterium longum genome sequence and the recent initiative to sequence additional strains are expected to open up a new era of comparative genomics in bifidobacterial biology. Moreover, the use of genomotyping allows a global comparative analysis of gene content between different bifidobacterial isolates of a given species without the necessity of sequencing many strains. Genomotyping provides useful information about the degree of relatedness among various strains of Bifidobacterium species and consequently can be used in a polyphasic identification approach. This review will deal mainly with the molecular tools described for bifidobacterial identification and the first insights into the underlying genetics involved in bifidobacterial physiology as well as genome variability.
Some simple formulae were obtained which enable us to estimate evolutionary distances in terms of the number of nucleotide substitutions (and, also, the evolutionary rates when the divergence times are known). In comparing a pair of nucleotide sequences, we distinguish two types of differences; if homologous sites are occupied by different nucleotide bases but both are purines or both pyrimidines, the difference is called type I (or "transition" type), while, if one of the two is a purine and the other is a pyrimidine, the difference is called type II (or "transversion" type). Letting P and Q be respectively the fractions of nucleotide sites showing type I and type II differences between two sequences compared, then the evolutionary distance per site is K = -(1/2) ln [(1-2P-Q) square root of 1-2Q]. The evolutionary rate per year is then given by k = K/(2T), where T is the time since the divergence of the two sequences. If only the third codon positions are compared, the synonymous component of the evolutionary base substitutions per site is estimated by K'S = -(1/2) ln (1-2P-Q). Also, formulae for standard errors were obtained. Some examples were worked out using reported globin sequences to show that synonymous substitutions occur at much higher rates than amino acid-altering substitutions in evolution.
Common marmosets (Callithrix jacchus) and cotton-top tamarins (Saguinus oedipus) (Callitrichidae, Primates) share a broadly similar diet of fruits, insects, and tree exudates. Common marmosets, however, differ from tamarins by actively gouging trees with their anterior teeth to elicit tree exudate flow. During tree gouging, marmosets produce relatively large jaw gapes, but do not necessarily produce relatively large bite forces at the anterior teeth. We compared the fiber architecture of the masseter muscle in tree-gouging Callithrix jacchus (n = 10) to nongouging Saguinus oedipus (n = 8) to determine whether the marmoset masseter facilitates producing these large gapes during tree gouging. We predict that the marmoset masseter has relatively longer fibers and, hence, greater potential muscle excursion (i.e., a greater range of motion through increased muscle stretch). Conversely, because of the expected trade-off between excursion and force production in muscle architecture, we predict that the cotton-top tamarin masseter has more pinnate fibers and increased physiological cross-sectional area (PCSA) as compared to common marmosets. Likewise, the S. oedipus masseter is predicted to have a greater proportion of tendon relative to muscle fiber as compared to the common marmoset masseter. Common marmosets have absolutely and relatively longer masseter fibers than cotton-top tamarins. Given that fiber length is directly proportional to muscle excursion and by extension contraction velocity, this result suggests that marmosets have masseters designed for relatively greater stretching and, hence, larger gapes. Conversely, the cotton-top tamarin masseter has a greater angle of pinnation (but not significantly so), larger PCSA, and higher proportion of tendon. The significantly larger PCSA in the tamarin masseter suggests that their masseter has relatively greater force production capabilities as compared to marmosets. Collectively, these results suggest that the fiber architecture of the common marmoset masseter is part of a suite of features of the masticatory apparatus that facilitates the production of relatively large gapes during tree gouging.