Brachypotherium (Perissodactyla,Rhinocerotidae) from the late Miocene of Samburu Hills,Kenya

Several isolated cheek teeth and mandibular specimens of Rhinocerotidae (Mammalia, Perissodactyla) from the upper Miocene Namurungule Formation in Samburu Hills, Kenya, are redescribed. Previously, these specimens had been identified as Chilotheridium pattersoni, Chilotheridium sp., Paradiceros mukirii, and Paradiceros sp. They are reidentified here as documenting the genus Brachypotherium based on their bucco-lingually broad molariform upper premolars with short crochet and flattened buccal walls on both upper and lower molars, the latter having a shallow external groove. Comparisons with other Brachypotherium species suggest that the present specimens belong to Brachypotherium sp. cf. B. minor. The presence of Brachypotherium in the Samburu Hills, at ca. 9.5 Ma, is concordant with the paleoenvironment (presence of lacustrine and river environments) known for this locality during the early late Miocene.     

Additional specimens of Diceros (Perissodactyla,Rhinocerotidae) from the Upper Miocene Nakali Formation in Nakali,central Kenya

An upper incisor and upper and lower cheek teeth of Rhinocerotidae from the Upper Miocene of Nakali in central Kenya are described. Those specimens are identified as Diceros sp. The present study confirms the presence of Diceros in sub-Saharan East Africa during Vallesian as noted by several studies. The present result and the fossil records of Diceros in Africa and Eurasia suggest that Diceros might have migrated to Eurasia from Africa by Vallesian, although more fossil records and detailed phylogenetic analysis of Diceros are needed to discuss this hypothesis.     

Molecular epidemiological survey of Babesia bovis,Babesia bigemina,and Babesia sp. Mymensingh infections in Mongolian cattle

Bovine babesiosis caused by Babesia species is an economically significant disease of cattle. Severe clinical babesiosis in cattle is caused by Babesia bovis, B. bigemina, and the recently discovered Babesia sp. Mymensingh. Mongolia is an agricultural country with a large cattle inventory. Although previous studies have detected active infections of B. bovis and B. bigemina in Mongolian cattle, only a few provinces were surveyed. Additionally, the endemicity of Babesia sp. Mymensingh in Mongolia remains unknown. We screened blood DNA samples from 725 cattle reared in 16 of the 21 Mongolian provinces using B. bovis-, B. bigemina-, and Babesia. sp. Mymensingh-specific PCR assays. The overall positive rates of B. bovis, B. bigemina, and Babesia sp. Mymensingh were 27.9% (n = 202), 23.6% (n = 171), and 5.4% (n = 39), respectively. B. bovis and B. bigemina were detected in cattle in all surveyed provinces; whereas Babesia sp. Mymensingh was detected in 11 of the 16 surveyed provinces. On a per province basis, the B. bovis- B. bigemina-, and Babesia sp. Mymensingh-positive rates were 5.9–52.0%, 9.1–76.3%, and 0–35.7%, respectively. In conclusion, this is the first report of Babesia sp. Mymensingh in Mongolia. In addition, we found that species of Babesia that are capable of causing bovine clinical babesiosis, including B. bovis, B. bigemina, and Babesia sp. Mymensingh, are widespread throughout the country.     

New Species of Boletellus Section Boletellus (Boletaceae,Boletales) from Japan,B. aurocontextus sp. nov. and B. areolatus sp. nov.

We describe and illustrate two new species of Boletellus section Boletellus, B. aurocontextus sp. nov. and B. areolatus sp. nov., which are generally assumed to be B. emodensis. In this study, we reconstructed separate molecular phylogenetic trees of section Boletellus using the nucleotide sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA, the largest subunit (RPB1) and the second-largest subunit (RPB2) of nuclear RNA polymerase II gene and mitochondrial cytochrome oxidase subunit 3 (cox3) gene. We also examined the morphologies of B. emodensis sensu lato (s.l.) and other related species for comparison. The molecular phylogenetic tree inferred from the sequences of nuclear DNA (ITS, and combined dataset of RPB1 and RPB2) indicated that three genetically and phylogenetically well-separated lineages were present within B. emodensis s.l. These three lineages were also distinguished on the basis of the molecular phylogenetic tree constructed using the sequences of mitochondrial DNA (cox3), suggesting distinct cytonuclear disequilibria (i.e., evidence of reproductive isolation) among these lineages. Therefore, these three lineages can be treated as independent species: B. aurocontextus, B. areolatus, and B. emodensis. Boletellus aurocontextus and B. areolatus are also distinct from B. emodensis by the macro- and microscopic morphologies. Boletellus aurocontextus is characterized by a pileus with bright yellow to lemon yellow context, which can be observed through a gap in the scales, and basidiospores with relatively large length (mean spore length, 21.4 μm; quotient of spore length and width, 2.51). In contrast, B. areolatus is characterized by a pileus with floccose to appressed thin scaly patches, a stipe with pallid or pale cream color at the upper half, and basidiospores with relatively small length (mean spore length, 16.5 μm; quotient of spore length and width, 1.80).     

Diverse Burkholderia Species Isolated from Soils in the Southern United States with No Evidence of B. pseudomallei

The global distribution of the soil-dwelling bacterium Burkholderia pseudomallei, causative agent of melioidosis, is poorly understood. We used established culturing methods developed for B. pseudomallei to isolate Burkholderia species from soil collected at 18 sampling sites in three states in the southern United States (Arizona (n = 4), Florida (n = 7), and Louisiana (n = 7)). Using multi-locus sequence typing (MLST) of seven genes, we identified 35 Burkholderia isolates from these soil samples. All species belonged to the B. cepacia complex (Bcc), including B. cenocepacia, B. cepacia, B. contaminans, B. diffusa, B. metallica, B. seminalis, B. vietnamiensis and two unnamed members of the Bcc. The MLST analysis provided a high level of resolution among and within these species. Despite previous clinical cases within the U.S. involving B. pseudomallei and its close phylogenetic relatives, we did not isolate any of these taxa. The Bcc contains a number of opportunistic pathogens that cause infections in cystic fibrosis patients. Interestingly, we found that B. vietnamiensis was present in soil from all three states, suggesting it may be a common component in southern U.S. soils. Most of the Burkholderia isolates collected in this study were from Florida (30/35; 86%), which may be due to the combination of relatively moist, sandy, and acidic soils found there compared to the other two states. We also investigated one MLST gene, recA, for its ability to identify species within Burkholderia. A 365bp fragment of recA recovered nearly the same species-level identification as MLST, thus demonstrating its cost effective utility when conducting environmental surveys for Burkholderia. Although we did not find B. pseudomallei, our findings document that other diverse Burkholderia species are present in soils in the southern United States.     

Genus Triticum L. taxonomy: the present and the future

The genetic relationships and diversity within the European and Asiatic Buxus species were analysed using AFLP, genome size analysis and chromosome counts. Based on these results two major clusters could be defined. One genetic cluster contained B. sempervirens and B. balearica, European species, and B. colchica, an Asiatic species but with leaf morphology similar to B. sempervirens. Species in this cluster were characterised by a genome size between 1.38 and 1.69?pg?2C^?1 and a chromosome number of 2n?=?2x?=?28 (diploid). Only four B. sempervirens cultivars within this cluster were triploid. A second cluster contained the Asiatic Buxus species B. microphylla, B. harlandii, B. hyrcana, B. myrica, B. henryi, B. bodinieri and B. wallichiana. Within this second genetic cluster three different ploidy levels could be observed. B. harlandii, B. hyrcana and nine B. microphylla cultivars were tetraploid (2n?=?4x?=?56) with a genome size of >2.5?pg?2C^?1. Fifteen other B. microphylla cultivars were triploid (2n?=?3x?=?42). The other Asiatic Buxus species, B. henryi, B. bodinieri and eight B. microphylla cultivars, were diploid with a genome size of ca. 1.5?pg?2C^?1.     

Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group

Bacillus anthracis, the causative agent of anthrax, is a potential source of bioterrorism. The existing assays for its identification lack specificity due to the close genetic relationship it exhibits to other members of the B. cereus group. Our comparative analyses of protein sequences from Bacillus species have identified a 24 amino acid deletion in a conserved region of the YeaC protein that is uniquely present in B. anthracis. PCR primers based on conserved regions flanking this indel in the Bacillus cereus group of species (viz. Bacillus cereus, B. anthracis, B. thuringiensis, B. mycoides, B. weihenstephnensis and B. pseudomycoides) specifically amplified a 282 bp fragment from all six reference B. anthracis strains, whereas a 354 bp fragment was amplified from 15 other B. cereus group of species/strains. These fragments, due to large size difference, are readily distinguished by means of agarose gel electrophoresis. In contrast to the B. cereus group, no PCR amplification was observed with any of the non-B. cereus group of species/strains. This indel was also used for developing a rapid pyrosequencing assay for the identification of B. anthracis. Its performance was evaluated by examining the presence or absence of this indel in a panel of 81 B. cereus-like isolates from various sources that included 39 B. anthracis strains. Based upon the sequence data from the pyrograms, the yeaC indel was found to be a distinctive characteristic of various B. anthracis strains tested and not found in any other species/strains from these samples. Therefore, this B. anthracis specific indel provides a robust and highly-specific chromosomal marker for the identification of this high-risk pathogen from other members of the B. cereus group independent of a strain's virulence. The pyrosequencing platform also allows for the rapid and simultaneous screening of multiple samples for the presence of this B. anthracis-specific marker.     

Unexpectedly high diversity of trypanosomes in small sub-Saharan mammals

The extremely species-rich genus Trypanosoma has recently been divided into 16 subgenera, most of which show fairly high host specificity, including the subgenus Herpetosoma parasitizing mainly rodents. Although most Herpetosoma spp. are highly host-specific, the best-known representative, Trypanosoma lewisi, has a cosmopolitan distribution and low host specificity. The present study investigates the general diversity of small mammal trypanosomes in East and Central Africa and the penetration of invasive T. lewisi into communities of native rodents. An extensive study of blood and tissue samples from Afrotropical micromammals (1528 rodents, 135 shrews, and five sengis belonging to 37 genera and 133 species) captured in the Central African Republic, Ethiopia, Kenya, Malawi, Mozambique, Tanzania, and Zambia revealed 187 (11.2%) trypanosome-positive individuals. The prevalence of trypanosomes in host genera ranged from 2.1% in Aethomys to 37.1% in Lemniscomys. The only previously known trypanosome detected in our dataset was T. lewisi, newly found in Ethiopia, Kenya, and Tanzania in a wide range of native rodent hosts. Besides T. lewisi, 18S rRNA sequencing revealed 48 additional unique Herpetosoma genotypes representing at least 15 putative new species, which doubles the known sequence-based diversity of this subgenus, and approaches the true species richness in the study area. The other two genotypes represent two new species belonging to the subgenera Ornithotrypanum and Squamatrypanum. The trypanosomes of white-toothed shrews (Crocidura spp.) form a new phylogroup of Herpetosoma, unrelated to flagellates previously detected in insectivores. With 13 documented species, Ethiopia was the richest region for trypanosome diversity, which corresponds to the very diverse environments and generally high biodiversity of this country. We conclude that besides T. lewisi, the subgenus Herpetosoma is highly host-specific (e.g., species parasitizing the rodent genera Acomys and Gerbilliscus). Furthermore, several newly detected trypanosome species are specific to their endemic hosts, such as brush-furred mice (Lophuromys), dormice (Graphiurus), and white-toothed shrews (Crocidura).     

Sexual and individual morphometric variation in Libycosaurus (Mammalia, Anthracotheriidae) from the Maghreb and Libya

Anthracotheres of late Middle Miocene and Late Miocene age have been described from several localities in northern Africa, all of them currently assigned to the genus Libycosaurus Bonarelli, although in several previous works they were assigned to Merycopotamus Falconer and Cautley, a considerably younger and specialised form from the Indian subcontinent which has quite different dental and cranial morphology. Three species of Libycosaurus have been named, but there has been some doubt about the morphometric variation within the various species, with some authors such as Gaziry (1987) placing the fossils from Sahabi (Latest Miocene, Libya) and Beglia (end Middle Miocene to basal Late Miocene, Tunisia) into the same species despite marked size differences, and others (Ducrocq et al., 2001) creating a species for a restricted sample of small specimens from Nementcha (Late Middle Miocene, Algeria), but which overlaps with the range of size variation of Beglia fossils. The aim of this paper is to examine the available samples in greater depth in order to understand the morphometric variation in these anthracotheres. It is confirmed that the Beglia sample is quite variable (Black, 1972), both morphologically and metrically, but it is concluded that it nevertheless belongs to a single species, because specimens from some of the localities within the Beglia Formation (e.g. Loc. 17 in the lower levels at Beglia) span the entire range of variation (Pickford, 1994). The sample from Nementcha cannot be distinguished from the Beglia sample on any consistent metric or morphological basis, but in general the specimens fall at the low end of the range of variation of the Beglia sample. It is thus likely that L. algeriensis is a synonym of L. anisae. The Sahabi and Chad samples (L. petrocchii), in contrast, fall above the known range of variation of the Beglia material in almost all metric features, but are close to it morphologically, and they are considered to represent a species distinct from the Beglia sample.     

Mark-release-recapture of males of Bactrocera cucurbitae and B. dorsalis (Diptera: Tephritidae) in two residential areas of Honolulu

This paper describes a mark-release-recapture study involving males of two economically important tephritid fruit flies (Diptera: Tephritidae), Bactrocera cucurbitae (Coquillett) and B. dorsalis (Hendel), conducted over a 2-year period in Honolulu, Hawaii. In each of two residential neighborhoods, we placed two traps, one baited with cue lure and the other with methyl eugenol (male attractants for B. cucurbitae and B. dorsalis, respectively), in a single tree. Dyed, mature males from recently established laboratory colonies were released 100 or 500 m from the traps along the four compass directions in both winter and summer seasons. In each neighborhood, a total of 5,600 flight-able males of each species were released 100 m from the traps (14 dates × 4 directions × 100 males/release) and 56,000 flight-able males of each species were released 500 m from the traps (14 dates × 4 directions × 1,000 males/release). Within each study area, the number of males trapped did not vary significantly with direction or season for either species for either the 100- or 500-m releases. Significantly higher numbers of B. dorsalis males were captured than B. cucurbitae males for both the 100-m (16 versus 8 males/release) and 500-m (7 versus 2 males/release) releases (average values computed over both study areas). Also, following their release, B. dorsalis males were, in general, trapped more quickly than B. cucurbitae males. Given the strong attractancy of methyl eugenol, trap captures for B. dorsalis were lower than expected, and possible explanations are discussed.     

Molecular characterization of the non-costate morphotypes of buliminid foraminifers based on internal transcribed region of ribosomal DNA (ITS rDNA) sequence data

We performed molecular phylogenetic analyses of four morphotypes of the benthic foraminiferal genus Bulimina (B. aculeata, B. marginata f. marginata, B. marginata f. denudata, and B. elongata) based on sequences of internal transcribed spacer region of ribosomal DNA (ITS rDNA). Six genetically distinct phylotypes were revealed by our phylogenetic analyses. The six phylotypes basically correspond to the fundamental morphotypes: clades A + B (B. aculeata); clade C (B. elongata); clade D (B. marginata f. denudata); clade E (B. marginata f. marginata genotype 1); and clade F (B. marginata f. marginata genotype 2). All six phylotypes are well distinguished, except phylotype B, which shows only little sequence divergence compared to clade A, possibly indicating that genetic differentiation is in progress. Morphological characters including the direction, placement, and shape of spines, the angle of undercutting of chamber periphery, and the roundness of the chambers were stable among specimens of each clade. In contrast, the length and density of spines, and chamber size, were variable within each clade. These intermediate morphological characters may reflect ecophenotypic variation. Our study clearly shows that the examined B. acuelata, elongata, and denudata morphospecies are genetically separated and that B. marginata is a species complex comprising several genotypes. A novel phylotype represent different morphotype compare to B. marginata f. marginata that can be distinguished based on differences of chamber angularity, the direction, placement, and shape of spines, and test dimensions.     

Cyanobacterial production of 1,3-propanediol directly from carbon dioxide using a synthetic metabolic pathway

Production of chemicals directly from carbon dioxide using light energy is an attractive option for a sustainable future. The 1,3-propanediol (1,3-PDO) production directly from carbon dioxide was achieved by engineered Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. Glycerol dehydratase catalyzing the conversion of glycerol to 3-hydroxypropionaldehyde in a coenzyme B₁₂-dependent manner worked in S. elongatus PCC 7942 without addition of vitamin B₁₂, suggesting that the intrinsic pseudovitamin B₁₂ served as a substitute of coenzyme B₁₂. The highest titers of 1,3-PDO (3.79±0.23 mM; 288±17.7 mg/L) and glycerol (12.62±1.55 mM; 1.16±0.14 g/L), precursor of 1,3-PDO, were reached after 14 days of culture under optimized conditions in this study.     

Capsalid monogeneans of fishes from the Seto Inland Sea,Japan: Description of Benedenia kobudai n. sp. parasitic on Semicossyphus reticulatus (Perciformes: Labridae)

The Seto Inland Sea, the largest inland sea in Japan, is a rich fishery with high biodiversity and productivity. Monogeneans have been studied for more than 120 years, and 58 nominal species have been recorded in the Seto inland Sea. This study provided DNA information on five species of Benedenia sensu lato (Capsalidae) from marine fishes from the sea, and one of them, Benedenia kobudai n. sp., is described from Semicossyphus reticulatus (Perciformes: Labridae). This new species differs from the other congeners by the hooded appearance between the anterior attachment organs, the morphology of the penis, the absence of the lobe near the vaginal pore and the common genital pore, the position of the vaginal pore, the germarium lying near the slightly hexagonal testes, the morphology of the haptor, and the shape and position of the hamuli. Phylogenetic analysis showed B. kobudai n. sp. in a separate clade from the other Benedenia species, B. epinepheli, B. hoshinai, B. sekii, and B. seriolae collected from the sea. Each of the newly provided DNA sequences (28S rDNA, ITS1‐5.8S-ITS2, cox1) of the above four species are based on specimens obtained from the type hosts and/or type localities and are considered important for future taxonomic re-examination and confirmation of the reliability of the registered sequences. Furthermore, these four species are important causes of fish diseases in aquaculture, and it is expected that information on the distribution, host range, and occurrence of fish diseases for each capsalid secured by molecular identification will be accumulated.The Life Science Identifier (LSID): urn:lsid:zoobank.org:pub:26C15D17-CFD1-450D-9FCE-EFFF692D2133.     

Trypanosoma lewisi: Enhanced resistance in naive lactating rats and their suckling pups

Lactating rats and their suckling offspring were shown to have a naturally occurring resistance to Trypansoma lewisi which was greater than that seen in normal adult rats and was not dependent on previous exposure to the parasite. In the case of the pups maximum protection was dependent on suckling being the sole source of nutriment and also on the peak of the parasitemia falling within the nursing period. Supplementary solid food reduced survival rates in concurrent Hemobartonella muris—T. lewisi infections: pups infected at 10 days of age and totally dependent on nursing showed 82.5% survival but only 6.6% survival when solid food was allowed to supplement milk. However, supplementary food did not reduce survival in pups infected with only T. lewisi. When pups were totally dependent upon nursing until the normal time of weaning (21 days of age), infection with H. muris—T. lewisi at the following days of age allowed the indicated mean survival rates: 10 days (82.5%), 20 (32.5%), 30 (14.3%), and 40 (100%). Infection with T. lewisi alone at the following days of age allowed the indicated mean survival rates: 10 days (100%), 15 (76%), 17 (52%), 20 (100%), and 30 (100%). Lactating rat serum agglutinated “adult forms” of T. lewisi, which correlated well with the observed sudden resolution of parasitemia in lactating animals at the time of the antigenic transition from “juvenile” to “adult” type. The “adult” stage parasitemia in suckling pups was selectively reduced when compared to that of nonlactating adult rats. The lactating rat serum factor could be passively transferred with lactating rat serum to animals already weaned.     

The spatial and vertical distribution of living (stained) benthic foraminifera from a tropical,intertidal environment,north Queensland,Australia

We investigated the distribution of living (stained) benthic foraminifera across a tropical, intertidal shoreline adjacent to Cocoa Creek, Queensland, Australia for the purpose of better understanding the nature of test production and ultimately fossil assemblage development within such environments. Short cores (up to 1 m) were collected during the wet and dry season, along an elevational gradient comprising non-vegetated intertidal mudflat and higher-intertidal mangrove forest environments. The distribution of stained specimens can be broadly delineated into assemblages characterising ‘upper mangrove’ (2.64–2.91 m above Lowest Astronomical Tide (LAT)) and ‘low mangrove-mudflat’ (1.62–2.18 m above LAT) environments. Agglutinated species were generally limited to upper mangrove stations. Calcareous species occurred within all of the intertidal environments examined but differ in their composition between upper and lower intertidal settings. Upper mangrove faunas were characterised by the agglutinated species Arenoparrella mexicana, Haplophragmoides wilberti, Miliammina fusca, Miliammina obliqua and Trochammina inflata and the calcareous species Helenina anderseni. Live (stained) assemblages at lower intertidal elevations were dominated by the calcareous species Ammonia aoteana, as well as Rosalina spp., Elphidium oceanicum, Triloculina oblonga, Ammonia pustulosa and Shackoinella globosa.     

Biological Control Activities of Rice-Associated Bacillus sp. Strains against Sheath Blight and Bacterial Panicle Blight of Rice

Potential biological control agents for two major rice diseases, sheath blight and bacterial panicle blight, were isolated from rice plants in this study. Rice-associated bacteria (RABs) isolated from rice plants grown in the field were tested for their antagonistic activities against the rice pathogens, Rhizoctonia solani and Burkholderia glumae, which cause sheath blight and bacterial panicle blight, respectively. Twenty-nine RABs were initially screened based on their antagonistic activities against both R. solani and B. glumae. In follow-up retests, 26 RABs of the 29 RABs were confirmed to have antimicrobial activities, but the rest three RABs did not reproduce any observable antagonistic activity against R. solani or B. glumae. According to16S rDNA sequence identity, 12 of the 26 antagonistic RABs were closest to Bacillus amyloliquefaciens, while seven RABs were to B. methylotrophicus and B, subtilis, respectively. The 16S rDNA sequences of the three non-antagonistic RABs were closest to Lysinibacillus sphaericus (RAB1 and RAB12) and Lysinibacillus macroides (RAB5). The five selected RABs showing highest antimicrobial activities (RAB6, RAB9, RAB16, RAB17S, and RAB18) were closest to B. amyloliquefaciens in DNA sequence of 16S rDNA and gyrB, but to B. subtilis in that of recA. These RABs were observed to inhibit the sclerotial germination of R. solani on potato dextrose agar and the lesion development on detached rice leaves by artificial inoculation of R. solani. These antagonistic RABs also significantly suppressed the disease development of sheath blight and bacterial panicle blight in a field condition, suggesting that they can be potential biological control agents for these rice diseases. However, these antagonistic RABs showed diminished disease suppression activities in the repeated field trial conducted in the following year probably due to their reduced antagonistic activities to the pathogens during the long-term storage in -70C, suggesting that development of proper storage methods to maintain antagonistic activity is as crucial as identification of new biological control agents.     

Morphology and Mitochondrial Genome of Fischoederius sp. 1 in Thailand

A rumen fluke Fischoederius elongatus is assigned to the type species of genus Fischoederius, family Gastrothylacidae. However, the mitochondrial sequences recently published are thought to be of inconsistent species, suggesting that several morphologically similar but genetically distinct species might be classified as Fischoederius elongatus. Thus, mentions of F. elongatus from South, Southeast, and East Asia might unintentionally refer to different species. The present work describes morphology and a full mitochondrial genome sequence of one of these species. The fluke specimens were collected from 2 infected cattle in Thailand. An interesting finding was the presence of a second tRNA-Asp gene next to a partial ND1 gene. It is suggested that these duplicated sequences are the remnants of non-reciprocal recombination events caused by inverted repeats located between ND2 and ND1 mitochondrial genes.     

Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group

Bacillus anthracis, the causative agent of anthrax, is a potential source of bioterrorism. The existing assays for its identification lack specificity due to the close genetic relationship it exhibits to other members of the B. cereus group. Our comparative analyses of protein sequences from Bacillus species have identified a 24 amino acid deletion in a conserved region of the YeaC protein that is uniquely present in B. anthracis. PCR primers based on conserved regions flanking this indel in the Bacillus cereus group of species (viz. Bacillus cereus, B. anthracis, B. thuringiensis, B. mycoides, B. weihenstephnensis and B. pseudomycoides) specifically amplified a 282 bp fragment from all six reference B. anthracis strains, whereas a 354 bp fragment was amplified from 15 other B. cereus group of species/strains. These fragments, due to large size difference, are readily distinguished by means of agarose gel electrophoresis. In contrast to the B. cereus group, no PCR amplification was observed with any of the non-B. cereus group of species/strains. This indel was also used for developing a rapid pyrosequencing assay for the identification of B. anthracis. Its performance was evaluated by examining the presence or absence of this indel in a panel of 81 B. cereus-like isolates from various sources that included 39 B. anthracis strains. Based upon the sequence data from the pyrograms, the yeaC indel was found to be a distinctive characteristic of various B. anthracis strains tested and not found in any other species/strains from these samples. Therefore, this B. anthracis specific indel provides a robust and highly-specific chromosomal marker for the identification of this high-risk pathogen from other members of the B. cereus group independent of a strain's virulence. The pyrosequencing platform also allows for the rapid and simultaneous screening of multiple samples for the presence of this B. anthracis-specific marker.