Infestation of an introduced host, the European green crab, Carcinus maenas, by a symbiotic nemertean egg predator, Carcinonemertes epialti

The recent introduction of the European green crab, Carcinus maenas, to the west coast of the U.S. has provided an opportunity for host transfer of the symbiotic nemertean egg predator, Carcinonemertes epialti, from its native shore crab host, Hemigrapsus oregonensis to the exotic C. maenas. Two surveys of C. maenas in Bodega Harbor, California, revealed that, in March 1995 prevalence of C. epialti on C. maenas was significantly lower than on H. oregonensis (11% versus 74%), but in November 1995 there was no significant difference between the 2 species (79% versus 98%). Only juvenile C. epialti were recovered from C. maenas in March 1995. However, in November 1995, ovigerous C. maenas were harboring actively feeding adult worms. Prevalence in both crab species significantly differed from March to November. Laboratory studies revealed that C. epialti fed and reproduced on eggs of C. maenas. The feeding rate of C. epialti on C. maenas eggs (2.5 eggs/trial) was not significantly different from that on H. oregonensis eggs (3.6 eggs/trial). Our findings suggest that this nemertean may have less host specificity than was previously thought. If C. epialti causes brood mortality of C. maenas in nature, it could potentially impact populations of this exotic crab.     

CYP330A1 and CYP4C39 enzymes in the shore crab Carcinus maenas: sequence and expression regulation by ecdysteroids and xenobiotics

Cytochrome P450 enzymes (CYP enzymes) catalyse important metabolic reactions of exogenous and endogenous substrates, including steroid hormones. Here, we report the first two CYP sequences from the shore crab, Carcinus maenas. Two complete cDNAs isolated from crab hepatopancreas encode CYP enzymes named CYP330A1, the first member of a new family, and CYP4C39. CYP330A1 is closest related to members of the CYP2 family (37.3% identical to mouse CYP2J6) and CYP4C39 is most identical to crayfish CYP4C15 (59.5%). CYP330A1 gene expression was induced in hepatopancreas of male green intermoult crabs by ecdysone and ponasterone A, but also by benzo(a)pyrene and phenobarbital. CYP330A1 induction was not observed in red crabs. The present results indicate that the CYP330A1 enzyme may be involved in ecdysteroid metabolism, presumably catabolism, and in the detoxification of environmental pollutants. Ecdysteroids or xenobiotics did not affect CYP4C39 gene expression. The fact that both ecdysteroids and xenobiotics affect CYP330A1 gene expression indicates that mutual interactions between chemical exposures and endocrine functions may exist in the shore crab.     

Continuous neurogenesis in the olfactory brain of adult shore crabs, Carcinus maenas

To scrutinize the common belief that the number of neurons in the CNS of adult decapod crustaceans stays constant, in spite of their dramatic postlarval increase in size, I counted olfactory projection neurons (OPNs) in the brains of differently-sized postlarval shore crabs, Carcinus maenas, and performed in vivo labeling of proliferating cells with 5-bromo-2'-deoxyuridine (BrdU) on brains of adults. The number of OPNs increases continuously throughout the postlarval life of shore crabs and approximately doubles from the very young to the oldest animals. Brain sections from adult crabs labeled with BrdU revealed ongoing proliferation of cells in the lateral soma cluster, which consists of OPN cell bodies, and in the cluster of somata of hemiellipsoid body local interneurons, which are the targets of the OPNs. Post-injection survival times from 5.5 to 120 h revealed a small but relatively constant number of labeled nuclei with neuronal morphology in both soma clusters of all specimens (31.3 +/- 9.5 S.D. nuclei per lateral cluster, n = 29; 20.1 +/- 4.5 S.D. nuclei per hemiellipsoid body cluster, n = 10). The labeled nuclei were located in a distinct proliferative zone in each cluster. There were significantly more labeled nuclei in both soma clusters after a prolonged post-injection survival time of 1 month (71.3 +/- 7.8 S.D. nuclei per lateral cluster, n = 4; 38.2 +/- 7.1 nuclei per hemiellipsoid body cluster, n = 6). In both soma clusters the labeled nuclei formed a compact group that was dislocated from the proliferation zone towards the outer edge of the cluster. In the proliferation zone of the lateral cluster histological stainings revealed cell bodies of typical neuronal shape that are slightly smaller and more intensely stained than the surrounding OPN somata. Some of these cell bodies were captured in various stages of mitosis. Collectively, these data indicate that continuous neurogenesis occurs in the central olfactory pathway of the brain of shore crabs throughout their entire adult life. This unexpected structural plasticity may enable long-lived decapod crustaceans to adapt to ever-changing olfactory environments.     

Association of marine archaea with the digestive tracts of two marine fish species

Recent studies have shown that archaea which were always thought to live under strict anoxic or extreme environmental conditions are also present in cold, oxygenated seawater, soils, the digestive tract of a holothurian deep-sea-deposit feeder, and a marine sponge. In this study, we show, by using PCR-mediated screening in other marine eukaryotes, that marine archaea are also present in the digestive tracts of flounder and grey mullet, two fish species common in the North Sea, in fecal samples of flounder, and in suspended particulate matter of the North Sea water column. No marine archaea could be detected in the digestive tracts of mussels or the fecal pellets of a copepod species. The archaeal 16S ribosomal DNA clone libraries of feces of flounder and the contents of the digestive tracts of grey mullet and flounder were dominated by group II marine archaea. The marine archaeal clones derived from flounder and grey mullet digestive tracts and feces formed a distinct cluster within the group II marine archaea, with 76.7 to 89. 8% similarity to previously described group II clones. Fingerprinting of the archaeal community of flounder digestive tract contents and feces by terminal restriction fragment length polymorphism of archaeal 16S rRNA genes after restriction with HhaI showed a dominant fragment at 249 bp, which is likely to be derived from group II marine archaea. Clones of marine archaea that were closely related to the fish-associated marine archaea clones were obtained from suspended particulate matter of the water column at two stations in the North Sea. Terminal restriction fragment length polymorphism fingerprinting of the archaeal community present in suspended particulate matter showed the same fragment pattern as was found for the archaeal community of the flounder digestive tract contents and feces. These data demonstrate that marine archaea are present in the digestive tracts and feces of very common marine fish. It is possible that the marine archaea associated with the digestive tracts of marine fish are liberated into the water column through the feces and subsequently contribute to the marine archaeal community of suspended particulate matter.     

Prevalence of two epicaridean isopods (Bopyridae and Entoniscidae) associated with the hermit crab Pagurus longicarpus Say, 1817 (Anomura) from the New Jersey coast (U.S.A.)

The epicaridean isopods Stegophryxus hyptius and Paguritherium alatum, parasitic in the hermit crab Pagurus longicarpus, are reported for the first time in New Jersey waters. Fourteen of 9,111 crabs (0.15%) were infested with the ectoparasitic S. hyptius, and 4 of 3,703 crabs (0.11%) were infected with the hemocoel-dwelling P. alatum. Crabs of both sexes were parasitized by each species. Prevalences of S. hyptius and P. alatum were considerably lower than those recorded over 50 yr ago in the Woods Hole region of Massachusetts.     

New brown chlorobacteria Prosthecochloris phaeoasteroidea nov. sp

Two strains of new photosynthetic bacteria were isolated from salt meromictic lakes Mogilnoye and Faro. The bacteria are abligate phototrophic anaerobic cultures which utilize H2S as an electron donor by oxidizing it to elementary sulphur and sulphates. Sulphur is liberated outside the cell. The cultures contain bacteriochlorophyll e and carotenoids of the isorenierathene type. Photosynthetic structures are represented by chlorobium-vesicles. According to these properties, the cultures belong to the Chlorobiaceae family. The bacteria have no gas vacuoles. The cells form 10--20 apophyses-prosthecae which is typical of the Prostheocochloris genus. The new strains differ from Prosthecochloris aestuarii by the brown colour of the cell suspension, the composition of pigments, and ecology. They are classed as a new species Prostheocochloris phaeoasteroidea nov. sp. Diagnosis of the new species is presented.     

Evolutionary relationships among members of the genus Chlamydia based on 16S ribosomal DNA analysis

Nucleotide sequences from strains of the four species currently in the genus Chlamydia, C. pecorum, C. pneumoniae, C. psittaci, and C. trachomatis were investigated. In vitro-amplified RNA genes of the ribosomal small subunit from 30 strains of C. pneumoniae and C. pecorum were subjected to solid-phase DNA sequencing of both strands. The human isolates of C. pneumoniae differed in only one position in the 16S rRNA gene, indicating genetic homogeneity among these strains. Interestingly, horse isolate N16 of C. pneumoniae was found to be closely related to the human isolates of this species, with a 98.9% nucleotide similarity between their 16S rRNA sequences. The type strain and koala isolates of C. pecorum were also found to be very similar to each other, possessing two different 16S rRNA sequences with only one-nucleotide difference. Furthermore, the C. pecorum strains truncated the 16S rRNA molecule by one nucleotide compared to the molecules of the other chlamydial species. This truncation was found to result in loss of a unilaterally bulged nucleotide, an attribute present in all other eubacteria. The phylogenetic structure of the genus Chlamydia was determined by analysis of 16S rRNA sequences. All phylogenetic trees revealed a distinct line of descent of the family Chlamydiaceae built of two main clusters which we denote the C. pneumoniae cluster and the C. psittaci cluster. The clusters were verified by bootstrap analysis of the trees and signature nucleotide analysis. The former cluster contained the human isolates of C. pneumoniae and equine strain N16. The latter cluster consisted of C. psittaci, C. pecorum, and C. trachomatis. The members of the C. pneumoniae cluster showed tight clustering and strain N16 is likely to be a subspecies of C. pneumoniae since these strains also share some antigenic cross-reactivity and clustering of major outer membrane protein gene sequences. C. psittaci and strain N16 branched early out of the respective cluster, and interestingly, their inclusion bodies do not stain with iodine. Furthermore, they also share less reliable features like normal elementary body morphology and plasmid content. Therefore, the branching order presented here is very likely a true reflection of evolution, with strain N16 of the species C. pneumoniae and C. psittaci forming early branches of their respective cluster and with C. trachomatis being the more recently evolved species within the genus Chlamydia.     

Use of green fluorescent protein to tag and investigate gene expression in marine bacteria

Two broad-host-range vectors previously constructed for use in soil bacteria (A. G. Matthysse, S. Stretton, C. Dandie, N. C. McClure, and A. E. Goodman, FEMS Microbiol. Lett. 145:87-94, 1996) were assessed by epifluorescence microscopy for use in tagging three marine bacterial species. Expression of gfp could be visualized in Vibrio sp. strain S141 cells at uniform levels of intensity from either the lac or the npt-2 promoter, whereas expression of gfp could be visualized in Psychrobacter sp. strain SW5H cells at various levels of intensity only from the npt-2 promoter. Green fluorescent protein (GFP) fluorescence was not detected in the third species, Pseudoalteromonas sp. strain S91, when the gfp gene was expressed from either promoter. A new mini-Tn10-kan-gfp transposon was constructed to investigate further the possibilities of fluorescence tagging of marine bacteria. Insertion of mini-Tn10-kan-gfp generated random stable mutants at high frequencies with all three marine species. With this transposon, strongly and weakly expressed S91 promoters were isolated. Visualization of GFP by epifluorescence microscopy was markedly reduced when S91 (mini-Tn10-kan-gfp) cells were grown in rich medium compared to that when cells were grown in minimal medium. Mini-Tn10-kan-gfp was used to create an S91 chitinase-negative, GFP-positive mutant. Expression of the chi-gfp fusion was induced in cells exposed to N'-acetylglucosamine or attached to chitin particles. By laser scanning confocal microscopy, biofilms consisting of microcolonies of chi-negative, GFP+ S91 cells were found to be localized several microns from a natural chitin substratum. Tagging bacterial strains with GFP enables visualization of, as well as monitoring of gene expression in, living single cells in situ and in real time.     

Diversity of free-living and attached bacteria in offshore Western Mediterranean waters as depicted by analysis of genes encoding 16S rRNA

In a previous study (S. G. Acinas, F. Rodríguez-Valera, and C. Pedrós-Alió, FEMS Microbiol. Ecol. 24:27-40, 1997), community fingerprinting by 16S rDNA restriction analysis applied to Mediterranean offshore waters showed that the free-living pelagic bacterial community was very different from the bacterial cells aggregated or attached to particles of more than about 8 micrometer. Here we have studied both assemblages at three depths (5, 50, and 400 m) by cloning and sequencing the 16S rDNA obtained from the same samples, and we have also studied the samples by scanning electron microscopy to detect morphology patterns. As expected, the sequences retrieved from the assemblages were very different. The subsample of attached bacteria contained very little diversity, with close relatives of a well-known species of marine bacteria, Alteromonas macleodii, representing the vast majority of the clones at every depth. On the other hand, the free-living assemblage was highly diverse and varied with depth. At 400 m, close relatives of cultivated gamma Proteobacteria predominated, but as shown by other authors, near the surface most clones were related to phylotypes described only by sequence, in which the alpha Proteobacteria of the SAR11 cluster predominated. The new technique of rDNA internal spacer analysis has been utilized, confirming these results. Clones representative of the A. macleodii cluster have been completely sequenced, producing a picture that fits well with the idea that they could represent a genus with at least two species and with a characteristic depth distribution.     

The evaluation by indirect immunofluorescence of the incidence of respiratory syncytial virus in acute respiratory disorders in infants]

Diversity based on ribosomal RNA gene-restriction endonuclease digest patterns was detected amongst forty-seven strains of Campylobacter made up of 38 strains of Campylobacter jejuni and 9 strains of Campylobacter coli. Restriction digests of chromosomal DNA prepared by treating with Hae III were probed with an oligonucleotide specific for Campylobacter 16S ribosomal RNA genes. Seventeen distinct hybridization patterns, each indicating the presence of 2-4 copies of the 16S rRNA gene are encoded in Campylobacter DNA. Differences in fragment patterns were observed not only between members of two species, but also between individual strains of the same species. Ribopattern fragments of 8.71, 7.56, 2.81 and 1.0 kb were characteristic of the majority of C. jejuni, whereas 7.59 and 4.68 kb fragments were commonly present in C. coli. In conclusion, Hae III ribotyping was even more discriminatory than the Penner serotyping of C. jejuni and C. coli, as strains of the same serotype were distinguished.     

Characterization of a defined 2,3,5, 6-tetrachlorobiphenyl-ortho-dechlorinating microbial community by comparative sequence analysis of genes coding for 16S rRNA

Defined microbial communities were developed by combining selective enrichment with molecular monitoring of total community genes coding for 16S rRNAs (16S rDNAs) to identify potential polychlorinated biphenyl (PCB)-dechlorinating anaerobes that ortho dechlorinate 2,3, 5,6-tetrachlorobiphenyl. In enrichment cultures that contained a defined estuarine medium, three fatty acids, and sterile sediment, a Clostridium sp. was predominant in the absence of added PCB, but undescribed species in the delta subgroup of the class Proteobacteria, the low-G+C gram-positive subgroup, the Thermotogales subgroup, and a single species with sequence similarity to the deeply branching species Dehalococcoides ethenogenes were more predominant during active dechlorination of the PCB. Species with high sequence similarities to Methanomicrobiales and Methanosarcinales archaeal subgroups were predominant in both dechlorinating and nondechlorinating enrichment cultures. Deletion of sediment from PCB-dechlorinating enrichment cultures reduced the rate of dechlorination and the diversity of the community. Substitution of sodium acetate for the mixture of three fatty acids increased the rate of dechlorination, further reduced the community diversity, and caused a shift in the predominant species that included restriction fragment length polymorphism patterns not previously detected. Although PCB-dechlorinating cultures were methanogenic, inhibition of methanogenesis and elimination of the archaeal community by addition of bromoethanesulfonic acid only slightly inhibited dechlorination, indicating that the archaea were not required for ortho dechlorination of the congener. Deletion of Clostridium spp. from the community profile by addition of vancomycin only slightly reduced dechlorination. However, addition of sodium molybdate, an inhibitor of sulfate reduction, inhibited dechlorination and deleted selected species from the community profiles of the class Bacteria. With the exception of one 16S rDNA sequence that had the highest sequence similarity to the obligate perchloroethylene-dechlorinating Dehalococcoides, the 16S rDNA sequences associated with PCB ortho dechlorination had high sequence similarities to the delta, low-G+C gram-positive, and Thermotogales subgroups, which all include sulfur-, sulfate-, and/or iron(III)-respiring bacterial species.     

The effect of background cuing on prey detection

Studies of prey detection have typically focused on how search image affects the capture of cryptic items. This study also considers how background vegetation influences cryptic prey detection. Blue jays, Cyanocitta cristata, searched digitized images for two Catocala moths: C. ilia, which is cryptic on oak, and C. relicta, which is cryptic on birch. Some images contained moths while others did not. The ability of blue jays to detect prey during repeated presentations of one prey type within a session was compared with their performance during randomly alternating presentations of both prey types within a session to examine search-image formation under two background conditions (informative and ambiguous). In the informative background condition, both trees in the image were of the same species and therefore, the background was a reliable indicator of which prey type might be present. In the ambiguous background condition, there was one tree of each species in the image and either prey type could be present. The results indicate that: (1) a search-image effect was observed only for the more cryptic prey type and only when the background was informative; (2) as accuracy on prey images (those with moths) increased, response latency remained unchanged; (3) performance on nonprey images (those without moths) was primarily determined by the difficulty of searching the background and not by the prey type in the accompanying prey images; and (4) search-image effects disappeared with extended practice. These results suggest that the ability to detect prey is influenced by background and that the presence of either multiple backgrounds or multiple prey types interferes with search-image formation. Copyright 1998 The Association for the Study of Animal Behaviour.     

Mixed function oxidase induction in Carcinus aestuarii. Field and experimental studies for the evaluation of toxicological risk due to Mediterranean contaminants

The aim of this study was to test and validate the use of mixed function oxidase (MFO) induction, in the crab Carcinus aestuarii, under experimental and field studies, for the evaluation of toxicological risk due to the main contaminants in the Mediterranean. Two different experiments were performed in the laboratory in order to identify the most suitable tissues for MFO studies in this species and the most suitable and sensitive MFO responses for evaluating chemical stress due to lipophilic contaminants. In order to validate this methodology in the field, two studies were carried out in two polluted Mediterranean lagoons: a transplant experiment in Orbetello Lagoon and an in situ experiment in Venice Lagoon. The following MFO responses were investigated in hepatopancreas and gills of the crabs: ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene hydroxylase (BPH) activities and reductase enzyme activities. The main results can be summarised as follows: midgut-gland and gills were confirmed to be useful for MFO tests; BPH activity in hepatopancreas was the most suitable and sensitive MFO response for evaluating chemical stress due to Mediterranean contaminants in laboratory and field studies; in the Orbetello Lagoon experiment, a statistically significant difference was found between sites subject to different human impact.     

Electron microscopical identification of the flagellate Spironucleus torosa (Hexamitidae) from burbot Lota lota (Gadidae) with comments upon its probable introduction to this freshwater host

Hexamitid flagellates from the rectum of the freshwater gadid burbot, from the river Glomma in southeastern Norway, were studied by scanning and transmission electron microscopy. The surface morphology of the flagellates was consistent with that of Spironucleus torosa, previously only found in the cod, haddock, and saithe, all marine gadids. Posteriolateral depressions with central protrusions were present in the flagellates, a feature reported from S. torosa only. Further, the microtubular cytoskeleton of the present species had the same general arrangement as in S. torosa, clearly different from what is described for other Spironucleus spp. It is therefore concluded that the present flagellate from burbot is this species. Any recent exchange of parasites between the marine hosts and burbot is believed to be only theoretically possible. The burbot became established in Norwegian rivers and lakes after the last ice age, some 7,000-8,000 yr ago, by following water courses across Sweden from the Baltic Sea. In the Baltic Sea there were, and still are, sympatric populations of burbot and cod, and the burbot is believed to have been infected before migrating westward.     

Phylogenetic differences between particle-associated and planktonic ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in the Northwestern Mediterranean Sea

The aim of this study was to determine if there were differences between the types of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria associated with particulate material and planktonic samples obtained from the northwestern Mediterranean Sea. A nested PCR procedure performed with ammonia oxidizer-selective primers was used to amplify 16S rRNA genes from extracted DNA. The results of partial and full-length sequence analyses of 16S rRNA genes suggested that different groups of ammonia-oxidizing bacteria were associated with the two sample types. The particle-associated sequences were predominantly related to Nitrosomonas eutropha, while the sequences obtained from the planktonic samples were related to a novel marine Nitrosospira group (cluster 1) for which there is no cultured representative yet. A number of oligonucleotide probes specific for different groups of ammonia oxidizers were used to estimate the relative abundance of sequence types in samples of clone libraries. The planktonic libraries contained lower proportions of ammonia oxidizer clones (0 to 26%) than the particulate material libraries (9 to 83%). Samples of the planktonic and particle-associated libraries showed that there were depth-related differences in the ammonia oxidizer populations, with the highest number of positive clones in the particle-associated sample occurring at a depth of 700 m. The greatest difference between planktonic and particle-associated populations occurred at a depth of 400 m, where only 4% of the clones in the planktonic library were identified as Nitrosomonas clones, while 96% of these clones were identified as clones that were related to the marine Nitrosospira species. Conversely, all ammonia oxidizer-positive clones obtained from the particle-associated library were members of the Nitrosomonas group. This is the first indication that Nitrosomonas species and Nitrosospira species may occupy at least two distinct environmental niches in marine environments. The occurrence of these groups in different niches may result from differences in physiological properties and, coupled with the different environmental conditions associated with these niches, may lead to significant differences in the nature and rates of nitrogen cycling in these environments.     

Nucleotide sequence of seed- and pollen-transmitted double-stranded RNA, which encodes a putative RNA-dependent RNA polymerase, detected from Japanese pear

The nucleotide sequence of the largest double-stranded (ds) RNA (named dsRNA1) of three species of seed- and pollen-transmitted dsRNA species detected from Japanese pear was analyzed, and one strand was found to contain a single long open reading frame (ORF) of 1434 nucleotides that encoded a putative polypeptide containing 477 amino acid residues with a molecular mass of 54.9 kDa. This polypeptide contained amino acid sequence motifs conserved in putative RNA-dependent RNA polymerases of RNA viruses. Attempts to visually identify or purify virus-like particles associated with the dsRNAs were unsuccessful. Slow sedimentation of the dsRNA fraction suggests that the dsRNAs may be unencapsidated. The concentration of dsRNAs in the host, Japanese pear, was about 16 times higher than that from a cryptic virus, radish yellow edge virus (RYEV). These results suggest that the dsRNAs were not from cryptic viruses. Partial nucleotide sequences of the two smaller dsRNAs (named dsRNAs 2 and 3) and two other dsRNAs (named dsRNAs 4 and 5) detected from only the Japanese pear cultivar (cv.) Akita Tazawa 3 Gou were analyzed, and encoded nearly the same amino acid sequence encoded by dsRNA1.     

Genetic analysis of inflammatory bowel disease in a large European cohort supports linkage to chromosomes 12 and 16

BACKGROUND & AIMS: Inflammatory bowel disease (IBD) is a complex disorder of unknown etiology. Epidemiological investigations suggest a genetic basis for IBD. Recent genetic studies have identified several IBD linkages. The significance of these linkages will be determined by studies in large patient collections. The aim of this study was to replicate IBD linkages on chromosomes 12 and 16 in a large European cohort. METHODS: Three hundred fifty-nine affected sibling pairs from 274 kindreds were genotyped using microsatellite markers spanning chromosomes 12 and 16. Affection status of the sibling pairs was defined as Crohn's disease (CD) or ulcerative colitis (UC). RESULTS: Nonparametric statistical analyses showed linkage for both chromosomes. Two-point results for chromosome 12 peaked at D12S303 (logarithm of odds [LOD], 2.15; P = 0.003) for CD and at D12S75 (LOD, 0.92; P = 0.03) for UC. Multipoint analyses produced a peak LOD of 1.8 for CD. Chromosome 16 showed linkage for CD at marker D16S415 (LOD, 1.52; P = 0.007). Multipoint support peaked above markers D16S409 and D16S411 (LOD, 1.7). CONCLUSIONS: These data are consistent with linkage of IBD to chromosomes 12 and 16. The replication of genetic risk loci in a large independent family collection indicates important and common susceptibility genes in these regions and will facilitate identification of genes involved in IBD.     

Phylogenetic affinity of a wide, vacuolate, nitrate-accumulating Beggiatoa sp. from Monterey Canyon, California, with Thioploca spp

Environmentally dominant members of the genus Beggiatoa and Thioploca spp. are united by unique morphological and physiological adaptations (S. C. McHatton, J. P. Barry, H. W. Jannasch, and D. C. Nelson, Appl. Environ. Microbiol. 62:954-958, 1996). These adaptations include the presence of very wide filaments (width, 12 to 160 microm), the presence of a central vacuole comprising roughly 80% of the cellular biovolume, and the capacity to internally concentrate nitrate at levels ranging from 150 to 500 mM. Until recently, the genera Beggiatoa and Thioploca were recognized and differentiated on the basis of morphology alone; they were distinguished by the fact that numerous Thioploca filaments are contained within a common polysaccharide sheath, while Beggiatoa filaments occur singly. Vacuolate Beggiatoa or Thioploca spp. can dominate a variety of marine sediments, seeps, and vents, and it has been proposed (H. Fossing, V. A. Gallardo, B. B. Jorgensen, M. Huttel, L. P. Nielsen, H. Schulz, D. E. Canfield, S. Forster, R. N. Glud, J. K. Gundersen, J. Kuver, N. B. Ramsing, A. Teske, B. Thamdrup, and O. Ulloa, Nature [London] 374:713-715, 1995) that members of the genus Thioploca are responsible for a significant portion of total marine denitrification. In order to investigate the phylogeny of an environmentally dominant Beggiatoa sp., we analyzed complete 16S rRNA gene sequence data obtained from a natural population found in Monterey Canyon cold seeps. Restriction fragment length polymorphism analysis of a clone library revealed a dominant clone, which gave rise to a putative Monterey Beggiatoa 16S rRNA sequence. Fluorescent in situ hybridization with a sequence-specific probe confirmed that this sequence originated from wide Beggiatoa filaments (width, 65 to 85 microm). A phylogenetic tree based on evolutionary distances indicated that the Monterey Beggiatoa sp. falls in the gamma subdivision of the class Proteobacteria and is most closely related to the genus Thioploca. This vacuolate Beggiatoa-Thioploca cluster and a more distantly related freshwater Beggiatoa species cluster form a distinct phylogenetic group.