Tursiops truncatus, T. aduncus or T. maugeanus; however the specific affinities of these animals is controversial and have recently been questioned. Historically ‘the southern Australian Tursiops’ was identified as unique and was formally named Tursiops maugeanus but was later synonymised with T. truncatus. Morphologically, these coastal dolphins share some characters with both aforementioned recognised Tursiops species, but they also possess unique characters not found in either. Recent mtDNA and microsatellite genetic evidence indicates deep evolutionary divergence between this dolphin and the two currently recognised Tursiops species. However, in accordance with the recommendations of the Workshop on Cetacean Systematics, and the Unified Species Concept the use of molecular evidence alone is inadequate for describing new species. Here we describe the macro-morphological, colouration and cranial characters of these animals, assess the available and new genetic data, and conclude that multiple lines of evidence clearly indicate a new species of dolphin. We demonstrate that the syntype material of T. maugeanus comprises two different species, one of which is the historical ‘southern form of Tursiops’ most similar to T. truncatus, and the other is representative of the new species and requires formal classification. These dolphins are here described as Tursiops australis sp. nov., with the common name of ‘Burrunan Dolphin’ following Australian aboriginal narrative. The recognition of T. australis sp. nov. is particularly significant given the endemism of this new species to a small geographic region of southern and south-eastern Australia, where only two small resident populations in close proximity to a major urban and agricultural centre are known, giving them a high conservation value and making them susceptible to numerous anthropogenic threats.Small coastal dolphins endemic to south-eastern Australia have variously been assigned to described species  Delphinids are the most ecologically diverse cetacean, occurring across a range of latitudes, in coastal and oceanic waters, and in estuarine and freshwater habitats Tursiops has been plagued with controversy with historically upwards of 20 species described, all synonymised with T. truncatusT. aduncus has been revalidated as the second Tursiops species, this based on morphological and mitochondrial DNA data Tursiops is polyphyletic Tursiops species recently suggested Tursiops species have been historically recognised as T. truncatusT. aduncus off eastern Australia, while in Western Australia aduncus and truncatus-type haplotypes are also present The genus Tursiops has been described for several decades Tursiops (as T. tursio). A single male specimen was captured by H.H. Scott in 1902 in the Cataract Gorge, Launceston, Tasmania. At the time, media reports, exhibition signage, and Scott's own handwritten notes  indicated that he believed it belonged to a distinct southern form of T. tursio. In 1914, a female specimen was obtained in the North Esk, Launceston, Tasmania. As stated, Scott and Lord Tursiops. Iredale and Troughton Tursiops maugeanus. Validity of the species has not been accepted by later authors and has been synonymised with T. truncatusT. maugeanus ‘holotype’ has been listed as unknown T. maugeanus and incorporated them into this contemporary analysis.In south-eastern Australia, morphological variation within T. truncatus and T. aduncusTursiops sp., southern Australian bottlenose dolphin (SABD) In the current study, morphology indicates two forms of ‘bottlenose’ dolphin in south-eastern Australia, a physically smaller coastal form in semi-enclosed water bodies, and a larger more robust ‘offshore’ form. Locations of beach-cast dolphins suggest these two forms are parapatric, at least across some of their range. The smaller coastal form has been noted as both Tursiops, Delphinus or Stenella species found world-wide. The average sequences divergence of the Victorian SABD to T. truncatus and T. aduncus  and Lagenorhynchus (4.5–6.4%) genera T. truncatus and T. aduncusLagenodelphis hosei (Fraser's Dolphin). Kingston et al. Tursiops species, but with the sister taxa Sousa chinensis (Indo-Pacific Humpback Dolphin). Unlike other recently recognised species, Orcaella heinsohniSotalia guianensisTursiops.Charlton et al. In 2004 a specialized Workshop on Cetacean Systematics was held to review cetacean taxonomy and provided criteria for species delimitation metapopulation lineages on different evolutionary trajectories, the farther along process of divergence, the larger the number of differences. In this Unified Species Concept, any property (line of evidence) that provides evidence of lineage separation is relevant in species delimitations including genetic, morphological, ecological or behavioural differentiation that characterizes the process of speciation” Currently there are numerous Species Concepts, each with underlying properties that represent thresholds crossed by diverging lineages, different subgroups of biologist advocate different species concepts but they all exhibit underlying conceptual unity T. maugeanus, and assess the taxonomic status of these animals. Consistent with the recommendations of the Workshop on Cetacean Systematics Since 2003 we have carried out extensive surveys, sampling and characterisation of ‘bottlenose’ dolphins from Victorian and Tasmanian coastal waters, using museum specimens, beach-cast strandings, live sightings and biopsies. In light of the confusion surrounding the taxonomy of these animals we use existing South-eastern Australia, encompassing coastal waters of Victoria and Tasmania . SoutherForty commonly used cranial measurements and tooth counts were taken from 44 specimens of ‘bottlenose’ dolphins from across Australia . Only adTursiops species were recognised as T. truncatus, QM specimens remain listed as either Tursiops sp. or T. truncatus. However T. aduncus is now known to be present in Queensland waters T. aduncus, and used the range (min-max) of cranial measures presented in Wang et al T. aduncus.As historically all Eighteen external morphometrics  were tak64,18 = 1.289, p = 0.28, male = 17 female = 9; external MANOVA, F11,6 = 1.056, p = 0.449, male = 11 female = 6). Data were standardized by converting the raw data to z-scores. Hierarchical cluster analysis (HCA) using Euclidean pair-group average, discriminant function analyses (DFA) and principal component analyse ((PCA) presented as supporting information only ystat v13 ast v1.94b Multivariate analyses of variance was used to test for sexual dimorphism in both cranial and external morphometrics datasets  , 0.25 M EDTA, saturated with NaCl, pH 7.5 Tooth samples were individually stored in sterilized Falcon tubes. Each tooth was sectioned and decontaminated by being submerged for 10 min in 12% sodium hypochloride b was undertaken following modified methods outlined in Charlton et al. Total genomic DNA was extracted from skin and tooth samples using a Puregene Tissue kit (Gentra Systems) following manufacturer's instructions, with modification for the teeth. Samples were analysed for quality and quantity of genomic DNA using a NanoDrop ND-1000 Spectrophotometer. PCR amplification and sequencing of a ∼450-bp fragment of the mtDNA control region and ∼1200-bp of cytochrome Tursiops maugeanusb gene and 417-bp region of the control region was carried out using seven and three sets of primers respectively, each amplified a 132–200-bp overlapping fragment . A model and parameters for the phylogenetic reconstruction were determined empirically using likelihood via Mega5 ega5 r Bayes 3.1.2 r Bayes were set to default. The analysis was run over 2 replicates to assure convergence on a topology. The Lagenorhynchus acutus sequence was used as outgroup  and Victorian State Government, Department of Sustainability and Environment (DSE) Wildlife Act 1975 Research Permit (Permit No: 10005013).Hierarchical cluster analysis was performed on all cranial variables for 44 specimens and showed three highly supported groups (cophenetic correlation of 0.8337) . Group 1A posteriori classifications were 100% correct. Canonical Discriminant function weighting identified rostrum measures RWM and RW60, width measure GPOW  and length measure TRIN  as important characters defining the groups   was used to determine whether cranial characteristics would distinguish the same groupings identified by the cluster analysis. The DFA scatterplot of canonical scores clearly show the three well separated ‘groups’ . A postee groups . Thirty-p<0.001) .Hierarchical cluster analyses performed on 14 variables using 17 specimens clearly showed two highly supported groups (cophenetic correlation of 0.747) .a posteriori individual group assignments corresponded exactly. Discriminant function weighting showed several length measures  and width of flukes (WFLU) as important characters defining the groups   . Again, e groups . Nine mep<0.001) .T. aduncus mtDNA types were found.Molecular analyses were limited to samples included in the cranial (n = 18) and external (n = 17) morphology analyses from south-east Australian samples . DNA data could not be obtained from QM skulls. From the 25 samples where DNA was available no b region was obtained from 18 samples representing both cranial and external morphology groups . Forty-eight fixed site differences were noted between the two species , when also including haplotypes previously reported in Charlton et al b region, T. australis sp. nov. showed less intra-specific variation (three variable sites) when compared to T. truncatus (13 variable sites). Genetic sequences from the current study have been deposited on GenBank  and southern form T. truncatus (Group 2)   & 2.T. maugeanus specimen (QVMAG#1365) and Group 2 incorporating the male T. maugeanus specimen (QVMAG#1360) (b and control region) place the female T. maugeanus (QVMAG 1365) within Tursiops australis sp. nov. and the male T. maugeanus holotype (QVMAG 1360) within T. truncatus  . MtDNA sQVMAG#136 and Grouruncatus .Tursiops australis sp. nov. clearly distinct from both Tursiops species, and in a monophyletic clade outside of any reported genera , Neighbour-joining (NJ) and Bayesian analyses showed d genera . MP and d genera . The trehinensis . Both MLThe electronic version of this document does not represent a published work according to the International Code of Zoological Nomenclature (ICZN), and hence the nomenclatural acts contained in the electronic version are not available under that Code from the electronic edition. Therefore, a separate edition of this document was produced by a method that assures numerous identical and durable copies, and those copies were simultaneously obtainable (from the publication date noted on the first page of this article) for the purpose of providing a public and permanent scientific record, in accordance with Article 8.1 of the Code. The separate print-only edition is available on request from PLoS by sending a request to PLoS ONE, Public Library of Science, 1160 Battery Street, Suite 100, San Francisco, CA 94111, USA along with a check for $10 (to cover printing and postage) payable to “Public Library of Science”.http://zoobank.org/”. The LSID for this publication is urn:lsid:zoobank.org:pub:9A469754-EFA0-499E-AF4C-772331B34025.In addition, this published work and the nomenclatural acts it contains have been registered in ZooBank, the proposed online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix “T. maugeanus syntypes together with specimens from numerous strandings over the past century, it is clear that the two syntype specimens of T. maugeanus comprise two different species , concurred with the undescribed species, SABD.In our quantitative and qualitative morphological and molecular comparisons of the designated southern form  species  & 5. In T. maugeanus we identified two alternative taxonomic treatments. One option was to designate the female as the lectotype of T. maugeanus and thus resurrect the name; with this option, the paralectotype male would simply be subsumed under T. truncatus. However, Scott's handwriting on the specimen label of the male, as well as his own extensive published and unpublished notes (held at QVMAG), make it clear that the species he envisaged was based on the male. Therefore, and given the current uncertain state of relationships within Delphininae T. truncatus sensu Scott and Lord T. maugeanus, in essence, assigned to the wrong form. The other option was to designate the male as the lectotype of T. maugeanus, and leave it as a questionable junior synonym of T. truncatus for now; with this option, the paralectotype female would then be left without an identity.To clarify the taxonomy of T. maugeanus as the appropriate available name for the southern form of T. truncatus, should it be found distinct from the northern. We anticipate that this is likely to occur, given the historical conclusions T. aduncus holotype T. aduncus, leaving the western Pacific/Southeast Asian form T. aduncus possibly requiring a new name After considerable consultation, we are convinced that the most conservative and stable approach is the second option above, thus leaving REVISED TAXONOMY OF TURSIOPS MAUGEANUS as TURSIOPS. TRUNCATUSOrder Cetacea Brisson, 1762Family Delphinidae Gray, 1821sensu LeDuc, 1999Subfamily Delphininae Tursiops Gervais, 1855Genus Tursiops maugeanus Iredale and Troughton, 1934Tursiops tursio, southern form male. – Scott and Lord 1919: 96, pl. XXIII–XXV [in part]Tursiops maugeanus - Iredale and Troughton 1934: 68, nom. nov. for T. tursio (southern form) Scott and Lord 1919 [in part].Tursiops truncatus - Möller et al. 2008: 676; Kemper 2004: 42; Perrin 2009.QVMAG 1360, Cataract Gorge, Launceston, Tasmania, Australia, 1902.T. maugeanus ( = southern form of T. tursio), but represents an entirely new form requiring a separate name.QVMAG 1365, Hobblers Bridge, North Esk River 5 km upstream from Tamar River, 11 November 1914; misidentified and does not in fact belong to Body large, robust ; with a short rostrum ; with tall and falcate dorsal fin ; with two-banded colouration dorsally slate grey-black, ventrally off-white; lacking a pale shoulder blaze and ventral spotting. Skull is large and robust , the rostrum is short  and wide across all measures , with shTursiops australis sp. nov.See below comparison with Tursiops maugeanus belong to two different species  is demonstrated to be a different species or subspecies from the northern form, the appropriate available name for the southern form would be T. maugeanus. The female paralectotype, which does not belong to T. maugeanus, is treated below.Concordant results from multiple independent data sets suggest that the syntype specimens of  species  & 5. Theruncatus  & 5. TheTAXONOMY OF NEW SPECIES.Order Cetacea Brisson, 1762Family Delphinidae Gray, 1821sensu LeDuc, 1999Subfamily Delphininae Tursiops Gervais, 1855Genus Tursiops australis sp. nov.urn:lsid:zoobank.org:act:54BA663A-BDE6-4E12-A9D2-84F6793EF4EATursiops tursio, southern form; female. - Scott and Lord 1919: 96, pl. XXIII–XXV [in part].Tursiops maugeanus - Iredale and Troughton 1937: 68, nom. nov. for T. tursio (southern form) Scott and Lord 1919 [in part].Tursiops sp. - Scarpaci et al. 2003: 342; Warren-Smith and Dunn 2006: 357.Victorian coastal bottlenose dolphin - Charlton et al. 2006: 173.Southern Australian Bottlenose Dolphin - Möller et al. 2008: 676; Owen et al. 2011.T. truncatus - Kingston et al. 2009:4.South Australian Tursiops truncatus - Ross and Cockcroft 1990: 124.Tursiops aduncus – Kemper 2004: 42.Tursiops truncatus : 75, pl. III.Not Tursiops tursio : 49.Not Tursiops aduncus Not australis, is in reference to the species link with Australia and is Latin for ‘southern’.Species name, Tursiops tursio by Scott and Lord, 1919, later named Tursiops maugeanus Iredale and Troughton, 1934. Repository location: Queen Victoria Museum and Art Gallery, Launceston, Tasmania, Australia.QVMAG 1365, Hobblers Bridge, North Esk River 5 km upstream from Tamar River, 11 November 1914; previously published as the female of the southern form of Monash MU210108, Beaumaris, Port Philip Bay, VIC, 21 Jan 2008; male. Monash MU230108, Point Henry, Geelong, VIC, 23 Jan 2008; male. MV C29579, Western Beach, Geelong, VIC, 14 Oct 1985; female. MV C29587, Kennedy's Point, Westernpoint Bay, VIC, 1 Jun 1992; male. MV C29667, Ocean Grove, VIC, 8 Jan 1987; female. MV C24944, Elwood, VIC, 2 Jun 1967; female. MV C28760, Sandringham, VIC, 13 Nov 1992; sex unknown. MV C29580, Murrells Beach, VIC, 17 Jan 1986; male. MV C29577, Safety Beach, VIC, 23 Jul 1985; female. MV C29586, Rippleside, VIC, 27 Jul 1991; male. MV C35986, Mitchell River, VIC, 4 Apr 2006; female. MV C35987, Hollands Landing, VIC, 21 Jul 2006. MV C35965, Lake Wellington, VIC, 14 Dec 2006; male. MV C35968, Poddy Bay, VIC, 30 Aug 2006; male. MV C35985, Blonde Bay, VIC, 1 Dec 2006; male. MV C35966, Lake Wellington, VIC, 14 Dec 2006; male. MV C36750, Paynesville, VIC, 4 Nov 2007; male. MV C29506, Sorrento, VIC, 16 Apr 1994; female. TMAG A1759, Marion Bay, TAS, 21 Feb 2003; unknown sex.North Esk River, 5 km upstream from Tamar River at Hobblers Bridge, Launceston, Tasmania , Port Phillip Bay and Gippsland Lakes, Victoria, Australia.Tursiops australis is smaller  than s.f. T. truncatus  . Dorsal fin is falcate like T. truncatus, c.f. the small triangular fin of T. aduncus. Tursiops australis has a tri-banded colouration grading conspicuously as follows: dark bluish-gray dorsally and on the sides of the head and body; light gray along the midline, extending as a pale shoulder blaze on the flank below the dorsal fin; and off-white ventrally, extending over the eye and above the flipper in some individuals; without ventral spotting (–3.02 m) , larger spotting .T. truncatus. Average skull length (CBL) is 493.58 mm (range 470–513 mm) smaller than that of s.f. T. truncatus (as above) and larger than T. aduncus . Across all measures the rostrum is wider and shorter than T. aduncus  . Teeth are long and conical, with older Gippsland Lakes and Tasmanian animals exhibiting substantial wear in the front and back teeth. The maxilla is flattened and smoothly transitional into the premaxilla toward the base of the rostrum, lacking the obvious the ‘pinched’ appearance of s.f. T. truncatus  . The ratruncatus .Tursiops australis differs from s.f. T. truncatus significantly at 58 diagnosable fixed base pairs across two mtDNA gene regions, 48 fixed site differences in a 1086-base sequence of the mtDNA cytochrome b region  meaning “name of a large sea fish of the porpoise kind” T. australis is in Port Phillip Bay where the Boonwurrung people have documented their existence for over 1000 years.We propose the common name ‘Burrunan Dolphin’ for T. australis occur in Victoria; Port Phillip Bay  show significant differences across multiple measures  and between the new species and T. aduncus (9.1%). Using mtDNA cytochrome b, Möller et al. T. truncatus. This is larger than between T. truncatus and T. aduncus (3.2%–5.8%) Lagenorhynchus obscurus and L. obliqidens (1.22%) Delphinus delphis and D. capensis (1.09%) Sotalia fluviatilis and S. guianensis (2.5%) Orcaella heinsohni and O. brevirostris (5.9%) T. australis and s.f. T. truncatus . Regardless of the DNA region used and phylogenetic analysis performed T. truncatus forms a separate clade from T. aduncusAs previously discussed, numerous studies have demonstrated that Tursiops in polyphyletic, with T. aduncus, T. truncatus and T. australis on three independent lineages. Using all three phylogenetic analyses, the placement of T. australis is outside of both Tursiops species, with a sister relationship to S. longirostris (using ML and NJ methods) and additionally L. hosei using Bayesian inferences. Möller et al. b suggests the same sister relationships.In this study we have also shown that Orcaella heinsohniSotalia guianensisTursiops is currently accepted as polyphyletic, Kingston et al. Tursiops species, despite the morphological similarities, and along with others, calls on a review of not only the genus Tursiops but of family Delphinidae Tursiops, more specifically the South African adunucus-type with the reported closeness to D. delphis, however no attempt was made at resolving the generic affinities. Given this current state of taxonomic uncertainty we believe that the most conservative approach at this time is to classify the new species in genus Tursiops, pending revision. We further believe that once revision of the Delphinidae is conducted, it is likely that this new species will be shown to represent a unique genus; if that is the case, we believe the genus name Tursiodelphis would be appropriate .Unlike other recently recognised species, T. australis, T. truncatus, T. aduncus and L. hosei (with species identification based on mtDNA regions). Of the five intron regions, four suggested no differentiation between the four species however one region (Del12) showed consistent species specific differences, defined by 3 variable sites . A possible explanation for this lack of differentiation may be due to the slower evolving nuclear regions, the rapid radiation of the delphinids . They include intron regions; CHRNA (283 bp) and POLA (330 bp) T. australis and s.f. T. truncatus is provided in Owen et al. T. australis (noted as SABD) was distinct from s.f. T. truncatus (noted as common bottlenose dolphin CBD), with s.f. T. truncatus having significantly lower values for δ13C and δ15N compared to T. australis. They conclude this distinction of the stable isotope signatures between the two species strongly indicates they forage in different areas and are likely to feed on different prey, thus providing an additional line of evidence for the recognition of T. australis.An additional line of evidence for separation of Tursiops truncatus and T. aduncus, of which we have demonstrated with clear and consistent evidence that it is neither. In addition, these dolphins have been ‘living under the eye’ in a well populated urban environment, have been the focus of multiple researchers and due to the multi-disciplinary approach taken in the manuscript we have been able to formally identify this species, thus highlighting the ‘need’ for other such studies to not ‘look in isolation’ of one line of evidence but to use a multiple disciplinary approach to assess the level of divergence.Whilst, as previously discussed, there is an urgent requirement to undertake a full review of the Delphinidae Family, this manuscript is an important step in this review. This new species has erroneously been ‘labelled’ T. australis would qualify for listing as a threatened species under the Australian Environment Protection and Biodiversity Conservation Act 1999 (EPBC Act) thus allowing immediate and directed conservation effort for further protection.The formal recognition of this new species is of great importance to correctly manage and protect this species, and has significant bearing on the prioritization of conservation efforts. This is especially crucial given it's endemism to a small region of the world, with only two small known resident populations and the proximity of those to major shipping ports, commercial and recreation fisheries, residential, industrial and agricultural stressors. Recognition of this new species opens the pathway that Figure S1Sampling locations of dolphins across Australia(TIF)Click here for additional data file.Figure S2Principal component analysis on cranial measures; scatter plot of Principal components 1 & 2(TIF)Click here for additional data file.Figure S3Principal component analysis on external morphology measures; scatter plot of Principal components 1 & 2(TIF)Click here for additional data file.Table S1Cranial measures(DOC)Click here for additional data file.Table S2External morphology measures(DOC)Click here for additional data file.Table S3Principal component analysis loadings of the first three Principal components (PC) on 34 cranial measures from 40 ‘bottlenose’ dolphin skulls(DOC)Click here for additional data file.Table S4Principal component analysis loadings of the first three Principal components (PC) for 11 external measures from 17 ‘bottlenose’ dolphins(DOC)Click here for additional data file.Table S5b gene and control region from Tursiops maugeanus syntype specimensPCR primers used to amplify mtDNA cytochrome (DOC)Click here for additional data file.Table S6GenBank accession numbers and species information for samples incorporated to phylogenetic analyses(DOC)Click here for additional data file.Table S7Discriminant function analysis loadings on 34 cranial measures from 40 ‘bottlenose’ dolphin skulls(DOC)Click here for additional data file.Table S8Discriminant function analysis loadings for 11 external measures from 17 ‘bottlenose’ dolphins(DOC)Click here for additional data file.Table S9Species classification overview based on different analyses and characters(DOC)Click here for additional data file.Table S10GenBank accession numbers of the sequences from this study(DOC)Click here for additional data file.Table S11Tursiops australis, Tursiops truncatus, Tursiops aduncus (current study) and from Tursiops aduncus holotype  and Tursiops adunucs Average cranial measures (mm) and tooth counts for (DOC)Click here for additional data file.


Laribacter hongkongensis is associated with community-acquired gastroenteritis and traveler's diarrhea. In this study, we performed an in-depth annotation of the genes in its genome related to the various steps in the infective process, drug resistance and mobile genetic elements.L. hongkongensis possessed a urease gene cassette, two arc gene clusters and bile salt efflux systems. For intestinal colonization, it possessed a putative adhesin of the autotransporter family homologous to those of diffusely adherent Escherichia coli (E. coli) and enterotoxigenic E. coli. To evade from host defense, it possessed superoxide dismutase and catalases. For lipopolysaccharide biosynthesis, it possessed the same set of genes that encode enzymes for synthesizing lipid A, two Kdo units and heptose units as E. coli, but different genes for its symmetrical acylation pattern, and nine genes for polysaccharide side chains biosynthesis. It contained a number of CDSs that encode putative cell surface acting (RTX toxin and hemolysins) and intracellular cytotoxins (patatin-like proteins) and enzymes for invasion (outer membrane phospholipase A). It contained a broad variety of antibiotic resistance-related genes, including genes related to β-lactam (n = 10) and multidrug efflux (n = 54). It also contained eight prophages, 17 other phage-related CDSs and 26 CDSs for transposases.For acid and bile resistance, L. hongkongensis genome possessed genes for acid and bile resistance, intestinal mucosa colonization, evasion of host defense and cytotoxicity and invasion. A broad variety of antibiotic resistance or multidrug resistance genes, a high number of prophages, other phage-related CDSs and CDSs for transposases, were also identified.The  Laribacter hongkongensis, a novel genus and species that belongs to the Neisseriaceae family of β-subclass of the Proteobacteria, was discovered from the blood and empyema pus of a patient with underlying alcoholic cirrhosis ..HelicobaL. hongkongensis strain HLHK9. They are related to β-lactam  (6 CDSs) of other bacterial species were found  that encodes the previously characterized class C β-lactamase [L. hongkongensis genome. They are both putative metallo-β-lactamases containing a metallo-β-lactamase superfamily domain which included two zinc ligand-binding sites essential for its hydrolytic function on the β-lactam ring  and one gene locus homologous to acrAD-tolC (LHK_02929-02931) of Escherichia coli were identified in the genome of L. hongkongensis. These three AcrRAB-TolC and the AcrAD-TolC multidrug efflux systems shared typical tripartite structure with other multidrug efflux systems in the RND family [acrR is a transcription regulator gene located upstream of the acrAB-tolC loci. As a multidrug efflux system with broad-substrate spectrum, AcrAB-TolC confers resistance to chloramphenicol, tetracyclines, erythromycin, trimethoprim, β-lactams, and other organic and inorganic antiseptic agents in E. coli [E. coli [L. hongkongensis is homologous to MdtABC-TolC system . MdtABC-TolC system in E. coli confers at least novobiocin and bile salt resistance in the bacterium. A uniqueness of this system is that MdtB and MdtC will form a heterodimer as a membrane efflux component in cooperation with membrane fusion protein MdtA and outer membrane channel protein TolC. [For Gram-negative bacteria, the efflux pumps that are associated with most clinically significant resistance to antibiotics are those of the RND family. In this family, three gene loci homologous to  E. coli ,84. AcrA[E. coli ,86. Anotin TolC. ,87 MoreoE. coli belonging to MFS were found in the genome of L. hongkongensis. One of them had an additional transcription regulator emrR gene (LHK_01376) in its upstream sequence. EmrAB-TolC system in E. coli confers nalidixic acid and other toxic novobiocin substances resistance to bacterium [emrR gene has been shown to lead to over-expression of the EmrAB pump and increased resistance to antimicrobial agents [L. hongkongensis is yet to be investigated. There are five other multidrug efflux proteins belonging to MFS  in the L. hongkongensis genome. One of them (LHK_00743) is a homolog to mdfA gene while another (LHK_02975) has high identities to bcr gene. mdfA encodes an MF-related protein, MdfA, which results in resistance to a diverse group of cationic and zwitterionic lipophilic compounds and antibiotics such as chloramphenicol and erythromycin when over-expressed in E. coli [bcr gene codes for an efflux protein which is associated with bicyclomycin resistance in E. coli [Two loci  homologous to emrAB-tolC system of acterium . Moreovel agents . However E. coli . bcr gen E. coli .L. hongkongensis. They are homologous to mdtJI  genes in E. coli which confers resistance to spermidine and, deoxycholate and sodium dodecyl sulfate at low level [mdtJI have to be co-expressed for functionality and it is suggested that MdtJI may function as a heterodimer or heterooligomer [Two adjacently located multidrug efflux genes (LHK_01384 and LHK_01385) of the SMR family were identified in the genome of ow level ,93. mdtJoligomer -94.L. hongkongensis. One of them (LHK_02533) is a homolog of multidrug efflux protein NorA from Staphylococcus aureus, which confers resistance to antibiotics of the quinolone class and various organic compounds [norA gene in S. aureus has resulted in 5- to 30-fold increase in susceptibility to norfloxacin [Two multidrug efflux genes of the MATE family (LHK_00466 and LHK_02533) were also discovered in the genome of ompounds ,96. MutaL. hongkongensis genome. A tripartite multidrug efflux system of the ABC transporter family composed of membrane transporter (LHK_02239), MFP (LHK_02240), and OMP (LHK_02241) was identified in the genome of L. hongkongensis. This system of proteins probably functions as a complex with composition resembling to that of RND family. Five other standalone putative ABC transporter genes  coding for multidrug efflux proteins were scattered over the L. hongkongensis genome. One (LHK_02949) of them possessed homology to msbA from E. coli, which is responsible for mediating the transport of the lipid A core of LPS to the outer membrane [E. coli MsbA in Lactococcus lactis which lacks LPS has been shown to significantly increase resistance to erythromycin [Six CDSs of the ABC transporter family related to multidrug resistance were identified in the membrane ,98. InteL. hongkongensis genome also encodes a number of other possible multidrug resistance-related genes. Among these, there are five marC-like genes , the expression of which was once believed to be associated with multidrug efflux system MarRAB in E. coli [marC did not increase antibiotic susceptibility on E. coli [Yersinia pestis and possessing an AbgT family domain was also identified in the genome of L. hongkongensis. AbgT protein family includes two transporter members, AbgT protein of E. coli and MtrF of N. gonorrhoeae [mtrCDE gene homologs were found in the genome of L. hongkongensis, the role and function of the AbgT family protein in L. hongkongensis remains to be elucidated.In addition to these five major families, the  E. coli . However E. coli . Therefoorrhoeae ,102. Mtrorrhoeae . Since nL. hongkongensis genome (Table ksgA have been documented [bacA gene (LHK_02940) encoding putative bacitracin resistance protein BacA was also identified. BacA protein confers bacitracin resistance to E. coli by catalyzing the dephosphorylation of undecaprenyl diphosphate (C55-PP) into C55-P, which is important in peptidoglycan synthesis. The conversion of C55-PP into C55-P is normally catalyzed by a specific phosphatase which is inhibited by bacitracin leading to halted peptidoglycan synthesis [E. coli has been shown to protect the bacteria against chromosome decondensation by camphor [crcB genes in L. hongkongensis genome, but only one copy in the closely related bacterium, C. violaceum, and none in N. gonorrheae or N. meningitidis genomes suggested that this is an important defense mechanism in L. hongkongensis. Since the L. hongkongensis strain, HLHK9, used for genome sequencing is susceptible to tetracycline (MIC = 0.5 μg/ml), the tetA gene previously identified in L. hongkongensis strains resistant to tetracycline is not found in the present genome [L. hongkongensis isolates from aquatic products in Guangzhou city, China [Six other CDSs with homologies to other drug resistance genes were identified in the me Table . A putatcumented -106. A bynthesis . The oth camphor . The pret genome . Recently, China . HoweverL. hongkongensis genome (genome size 3.16 Mbp) contains a total of eight putative prophages named LhP1 to LhP8, the positions of which are shown in Figure C. violaceum (genome size 4.75 Mbp) (GenBank accession no. AE016825), 1 to 3 in N. meningitidis (genome size 2.14 to 2.27 Mbp)  and 6 in N. gonorrhoeae (genome size 2.15 to 2.23 Mbp)  using the same parameters for prophage prediction by Prophage Finder, suggested that this is an important mechanism for acquisition and exchange of genetic materials in L. hongkongensis. While N. meningitides and C. violaceum cause mainly meningitis and invasive infections respectively that can lead to fatal septicemia, N. gonorrheae and L. hongkongensis were mainly isolated from human genital and gastrointestinal tract respectively. Interestingly, the presence of apparently high number of prophages also in N. gonorrhoeae is in line with our previous observation that horizontal gene transfer was particularly frequent among bacteria residing in human gastrointestinal and probably genital tract [The al tract , suggestL. hongkongensis genome. LhP1 contains 34 phage-related CDSs. Analysis of these CDSs indicated that LhP1 is likely a P2-like phage, as 29 of its 34 phage-related CDSs were most similar to CDSs in P2-like prophages  in the order Caudovirales (tailed phages) [Myoviridae and Siphoviridae (phages with long non-contractile tails).Bacteriophage LhP1 is composed of 47 CDSs, accounting for 31,318 bp with G+C content 63.07%, close to the G+C content of the s Figure . A P2-li phages) . Other CC. violaceum (CvP1), Haemophilus influenzae and N. meningitidis. There are also other CDSs similar to other phage genes of lambda- and P2-like phages.Bacteriophage LhP2 is composed of 32 CDSs, accounting for 26,141 bp with G+C content 64.81%. Analysis of its CDSs indicated that LhP2 is likely a Mu-like phage, with 10 of the 25 phage-related CDSs most similar to CDSs in Mu-like phages of L. hongkongensis genome. It is composed of 19 CDSs, accounting for 11,169 bp with G+C content 58.70%, lower than that of the host genome (62.35%), reflecting its heterologous origin. Of the 19 CDSs, 14 were phage-related CDSs with similarity to genes of BPP-1-, lambda- and epsilon15-like phages and other unclassified phages, indicating its genetic complexity. Further studies are required if this relatively small prophage is a functional tailed phage.Bacteriophage LhP3 is the smallest prophage in the Bordetella phage BPP-1. Other phage related genes resemble those of P4-, P2-, P22- and episolon15-like phages and unclassified phages of Siphoviridae and Myoviridae.Bacteriophage LhP4 is composed of 36 CDSs, accounting for 34,375 bp with G+C content 58.78%, also lower than that of the host genome, indicating its heterologous origin. Of the 23 phage-related CDSs, 14 possessed similarity to genes of L. hongkongensis genome. Composed of 64 CDSs, it accounts for 43,997 bp with G+C content 59%, lower than that of the host genome. Of the 32 phage-related CDSs, 9 possessed homologies to genes of Mu-like phages, 7 even possessed homologies to genes of lambda-like phages. The other phage-related CDSs are most closely related to those of various phages including those belonging to Podoviridae (phages with short tails), Myoriviridae and Siphoviridae.Bacteriophage LhP5 is the largest prophage identified in the Bordetella bronchiseptica and Bordetella avium. Of these 25 CDSs, 12 possessed homologies to genes of unclassified phages belonging to Siphoviridae and 5 to lambda-like phages.Bacteriophage LhP6 is composed of 31 CDSs, accounting for 21,918 bp with G+C content of 62.04%. The 25 phage-related CDSs exhibit similarity to phage genes of N. meningitidis, Burkholderia, and C. violaceum genes of Mu-like phages, and others to those of unclassified phages, lambda-, P22-, and BPP-1-like phages.Bacteriophage LhP7 is composed of 31 CDSs, accounting for 19,992 bp with the lowest G+C content of 55.59% among the eight prophages, suggesting a heterologous origin. Of the 18 phage-related CDSs, 4 exhibits similarity to phage genes of Myoviridae, Siphoviridae and Mu-like phages. In fact, LhP1 and LhP8 are highly similar with the exception of a few CDSs, with most of their CDSs exhibiting similarity to phage proteins found in other gram-negative bacteria including Salmonella, Burkholderia, Yersinia, and Shigella species. Their gene organizations are also highly similar to P2 phage  and LHK_01281 (ORFa), with 38-bp inverted repeats with six mismatches. The second insertion sequence is relatively short in length, with 603 bp containing two ORFs, LHK_02311 and LHK_02312 (ORFa) and 50-bp inverted repeats with ten mismatches. The G+C content of both putative insertion sequences are lower (57.4% and 54.89% respectively) than that of the L. hongkongensis genome, suggestive of heterologous origin.There are 26 CDSs coding for transposases in the me Table . Fourteeiolaceum . The traL. hongkongensis genome possessed genes and gene cassettes for acid and bile resistance, colonization of the intestinal mucosa, evasion of host defense and cytotoxicity and invasion. In addition, a broad variety of antibiotic resistance or multidrug resistance genes, a high number of prophages, together with other phage-related CDSs and CDSs coding for transposases, were also identified.The L. hongkongensis genome were annotated as described in our previous publication and classified functionally according to the Clusters of Orthologous Groups (COG) methodology [http://bioinformatics.uwp.edu/~phage/ searches [L. hongkongensis HLHK9 against the ISfinder database http://www-is.biotoul.fr/is.html[CDSs identified in the hodology . CDSs behodology  and keywsearches . The genr/is.html and inver/is.html. Manual r/is.html.Chromobacterium violaceum phage; DAEC: Diffusely adherent Escherichia coli; DNA: Deoxyribonucleic acid; dTDP: Deoxythymidine diphosphate; EMBOSS: European Molecular Biology Open Software Suite; ETEC: Enterotoxigenic Escherichia coli; IS: Insertion sequence; Kdo: Keto-deoxyoctulonate; LhP: Laribacter hongkongensis prophage; LPS: Lipopolysaccharide; MATE: Multidrug and Toxic compound Extrusion; Mbp: Mega base pairs; MF: Major facilitator; MFP: Membrane fusion protein; MFS: Major Facilitator Superfamily; MIC: Minimum inhibitory concentration; OMP: Outer membrane (channel) protein; OMPLA: Outer membrane phospholipase A; ORF(s): Open reading frame(s); PBP(s): Penicillin-binding protein(s); RND: Resistance nodulation division; RTX: Repeats in toxin; SMR: Small Multidrug Resistance family.ABC: ATP-Binding Cassette; ATP: Adenosine triphosphate; BLAST: Basic Local Alignment Search Tool; bp: Base pair; C55-P: Undecaprenyl pyrophosphate; C55-PP: Undecaprenyl diphosphate; CD14: Cluster of differentiation 14; CDS(s): Coding sequence(s); COG(s): Clusters of orthologous group(s); CvP: The authors declare that they have no competing interests.PCYW, KYY and SKPL designed and supervised the study. GKMW, AKLT JLLT, and RYYF annotated the genome. HT performed bioinformatics analysis. SKPL, GKMW, AKLT and PCYW drafted the manuscript. All authors corrected the manuscript. All authors read and approved the final manuscript.


Vibrio fluvialis among 400 nonagglutinating Vibrio spp. isolated from patients with diarrhea in Kolkata, India. For 43 patients, V. fluvialis was the sole pathogen identified. Most strains harbored genes encoding hemolysin and metalloprotease; this finding may contribute to understanding of the pathogenicity of V. fluvialis.We identified 131 strains of  Vibrionaceae cause diarrheal disease; among these, Vibrio cholerae O1/O139 and V. parahaemolyticus are responsible for several epidemics and pandemics  vibrios.Many members of the family V. fluvialis has caused sporadic cases and outbreaks of diarrhea in several countries , heat-stable enterotoxin (stn), type 3 secretion system , cholera toxin (ctxA), toxin co-regulated pilus (tcpA), thermostable direct-hemolysin (tdh), TDH-related hemolysin (trh), V. fluvialis hemolysin (VFH), and metalloproteases, according to published methods , according to Clinical and Laboratory Standards Institute criteria . The isolation rate of V. fluvialis gradually increased from 0.7% in 2002 to 2.2% in 2009  were identified as the sole pathogen; the remaining 88 (67%) were isolated as a co-pathogen with either V. cholerae, V. parahaemolyticus, E. coli, Shigella spp., parasites, or enteric viruses (data not shown). Among the mixed infections, V. fluvialis with V. cholerae was isolated most often (17%), followed by V. fluvialis and V. parahaemolyticus (8%). The presence of Vibrio spp. as mixed pathogens indicates that these patients likely acquired the infection from contaminated water or food. We analyzed the date of admission and place from where the patients resided and found no evidence for clusters of infection or small outbreaks caused by V. fluvialis.Among the 400 isolates presumptively identified NAG vibrios, multiplex PCR confirmed 131 and 269 strains (each strain representing a case) as  in 2009 . Of the V. fluvialis infection was much more often detected in adults (73%) than in children <5 years of age (27%). Clinical symptoms of sole infection caused by V. fluvialis were similar to that of cholera: watery diarrhea (86%), severe dehydration status (28%), and abdominal pain (12%)  . SeveralV. fluvialis strains were negative for the virulence genes commonly reported in V. cholerae and V. parahaemolyticus, but >90% were positive for genes encoding VFH and metalloproteases. More than 80% of the strains expressed hemolysin against rabbit and sheep red blood cells. Hemolysin is a widely distributed virulence factor in most pathogenic Vibrio spp. Metalloprotease produced by V. fluvialis is related to hemagglutination proteases of V. vulnificus, which enhances permeability and hemorrhagic activities , streptomycin (85%), furazolidone (85%), and sulfamethoxazole/trimethoprim (70%) -Ib-cr), which reduces ciprofloxacin activity  . About hV. fluvialis strains exhibited distinct NotI restriction profiles in the denrogroam analysis, at least 4 major clades were identified (Although the entified . Clades entified .V. fluvialis among patients with acute diarrhea patients in Kolkata. The expression of cytotoxic activity and hemolysin may contribute to understanding the pathogenicity of V. fluvialis. Further epidemiologic studies are necessary to elucidate the public health importance of V. fluvialis–mediated diarrhea.Our results demonstrate an emerging trend of prevalence of Vibrio fluvialis strains to determine ability to lyse rabbit erythrocytes and cytotoxic effect on Chinese hamster ovary and HeLa cells and antimicrobial drug resistance of V. fluvialis.Results of assays of clinical


Pregabalin is used in the treatment of postherpetic neuralgia, diabetic neuropathic pain, partial seizures, anxiety disorders and fibromyalgia. Recognized adverse effects associated with its use include cognitive impairment, somnolence and dizziness. Heart failure associated with pregabalin has been described, however the strength of this association has not been well characterized. To examine this further, we will conduct a systematic review of the risk of heart failure and edema associated with use of pregabalin.We will include all studies  that examine the use of pregabalin compared to placebo, gabapentin or conventional care. Our primary outcome is heart failure and the secondary outcomes include edema and weight gain. We will search electronic databases , and grey literature sources  to identify relevant studies. To ensure literature saturation, we will contact drug manufacturers, conduct forward citation searching, and scan the reference lists of key articles and included studies. We will not restrict inclusion by language or publication status.Two reviewers will screen citations (titles and abstracts) and full-text articles, conduct data abstraction, and appraise risk of bias. Random-effects meta-analysis will be conducted if the studies are deemed heterogeneous in terms of clinical, statistical and methodological factors but still suitable for meta-analysis.The results of this review will assist physicians to better appreciate pregabalin’s risk for edema or congestive heart failure and will be pertinent to the thousands of patients worldwide who are administered this medication.Our protocol was registered in the PROSPERO database (CRD42012002948). Pregabalin, a structural analogue to gamma-aminobutyric acid (GABA), is a new medication that is widely prescribed for chronic pain syndromes such as postherpetic neuralgia and diabetic neuropathic pain, as well as partial seizures, anxiety disorders and fibromyalgia . It is aPregabalin’s known adverse effects include cognitive impairment, somnolence and dizziness . Post-maThe risks associated with pregabalin may be better studied by examining observational studies, which include greater numbers of patients with comorbid conditions – studies not included in previous reviews . We hypoThis research protocol was developed by our team with expertise in geriatric medicine, clinical pharmacology, systematic review methodology, statistics and library science. Our protocol was registered in the PROSPERO database (CRD42012002948).e.g., language publication status or year of dissemination)We will include any study report that examines congestive heart failure, edema or weight gain among adult patients (age ≥18 years) newly prescribed pregabalin compared to gabapentin, placebo or standard medical care. Studies will be included regardless of publication status or language of dissemination. We will exclude qualitative studies but all other study designs will be included. The PICOST criteria are as follows:Patients – : Adults ≥18 years; Intervention – : Pregabalin (any dosage and duration of use); Comparator – : Gabapentin  placebo or conventional medical care; Outcome – : Heart failure (primary) edema weight gain alone or with edema (secondary); Studies – : Clinical trials , quasi-experimental studies (controlled before-after studies and interrupted time series), observational studies , case series/reports, safety bulletins and primary surveillance data from drug regulatory agencies; Time – : No restrictions will be imposed based upon study duration; Other – : No other restrictions will be imposed  and conference abstracts  for difficult to locate or unpublished  literature. The following Regulatory Authority safety alerts will be searched: Food and Drug Administration MedWatch (United States), European Medicines Evaluation Agency's European Public Assessment Reports, Medicines and Healthcare Products Regulatory Agency (United Kingdom), Australia Adverse Drug Reactions Bulletin, Health Canada MedEffect and the Canadian Adverse Drug Information System. We will also search the World Health Organization Uppsala Monitoring Centre.An experienced librarian (LP) will conduct the literature search and the search strategy will be peer reviewed by another librarian using the Peer Review of Electronic Search Strategies (PRESS) checklist . We willThe eligibility criteria will be pilot-tested on a random sample of 50 citations, which will be screened by the entire team. A kappa statistic will be calculated to measure inter-rater reliability and screening will only commence when ≥60% agreement is achieved . Using ti.e., companion reports). To identify studies that generate multiple reports (duplication bias), we will record the authors' names, study location and setting, dose and frequency of pregabalin administration (intervention), number of participants and their baseline demographic data, and date and duration of the study. Once identified, we will link these reports. We will consider the report with the longest duration of follow-up or primary outcome of interest as the major publication and the rest will be considered companion reports which provide supplementary information.Some of the included study reports might be studies examining pregabalin among the same patient population , we wilA data abstraction form will be developed and amended following a pilot-test on a 5% random sample of the included studies by all reviewers. Two reviewers will subsequently perform all data abstraction in duplicate. Conflicts will be resolved by discussion or, if necessary, a third reviewer will be involved. We will attempt to contact study authors to verify data, as necessary. The anticipated data that will be collected are included in Appendix B.For randomized clinical trials, we will use the Cochrane Collaboration's 5.1.0 risk of bias tool. This 7-item tool assesses for selection bias, performance bias, detection bias, attrition bias, and reporting bias .For non-randomized studies and observational studies, a single risk of bias tool has not been validated . As suchThe following are variables that are confounders for pregabalin and the risk for edema or heart failure:1. Demographics: Age, gender, sociodemographic status/income2. Comorbidities: Diabetes, hypertension, dyslipidemia, congestive heart failure, cardiovascular disease, cerebrovascular disease, renal disease, seizure disorder, number of medications per year, burden of illness scales 3. Medications that affect fluid balance: Loop diuretics4. Cardiovascular medications: Renin-angiotensin antagonists , negative chronotropes , statins, antiarrhythmics, antiplatelet agents, anticoagulants5. Medications that can exacerbate or trigger congestive heart failure: Non-steroidal anti-inflammatory drugs, steroids, thiazolidinediones6. Medications associated with severe neuropathic pain: Opioids7. Medications associated with other indications for pregabalin: Anticonvulsants, benzodiazepines, antidepressantse.g., odds ratio, relative risk).We will record which studies adjusted for which variables, and their crude and adjusted measures of effect size :1. gamma-Aminobutyric Acid/aa [Analogs & Derivatives]2. pregabalin.tw.3. lyrica.tw.4. "3-isobutyl GABA".tw.5. pregablin.mp.6. CI-1008.tw.7. S1731_Selleck.tw.8. "3-(aminomethyl)-5-methylhexanoic acid".tw.9. 148553-50-8.rn. [CAS Registry Number]10. "3 isobutyl 4 aminobutyricacid".tw.11. "3 isobutylgaba".tw.12. "4 amino 3 isobutylbutyric acid".tw.13. "pd 144723".tw.14. or/1-1315. exp Adult/ 16. adult.mp.17. Middle Aged/18. age$.tw.19. or/15-1820. 14 and 1921. exp Animals/ not  [ removing animal studies ]22. 20 not 21The following data categories will be collected:1. Patient characteristics:1. Total number 1. Setting1. Diagnostic criteria1. Age 1. Gender 1. Country1. Co-morbidities1. Socio-demographics1. Ethnicity1. Date of study2. Study characteristics:1. Report ID (created by review author)1. Citation and contact details1. Confirm eligibility for review1. Reason for exclusion1. Study design1. Country1. Setting 1. Publication status1. Intervention and comparator descriptions 1. Allocation to groups vs. components)1. Prospective 1. Funding source1. Key conclusions of the study authors1. Miscellaneous comments from study authors3. Quality/Risk of bias:1. Cochrane 5.1 collaboration's tool for assessing risk of bias categories 1. Newcastle-Ottawa Scale for assessing risk of bias categories (cohort and case–control studies)1. Effective Practice and Organization of Care (EPOC) for assessing risk of bias 1. Naranjo Adverse Drug Reporting Probability Scale (case reports/series)1. McMaster Quality Assessment Scale for Harms Study (McHarm)4. Adverse events 1. Diagnostic criteria1. Number of events in intervention and control/comparison groups1. Sample size1. Frequency1. Relative risk 1. Odds ratio (cohort studies)1. Severity1. Resulted in withdrawals1. Collected at follow-up (frequency of follow-up)1. Collected by patient diary or checklist or spontaneous reportingvs. late withdrawal1. Early 1. Other adverse events reportedThe authors declare that they have no conflicts of interest.JMH conceived the study, designed the study, helped develop the search strategies, registered the protocol and helped write the draft protocol. ACT helped design the review methods, and edited the draft protocol. LP helped develop the search strategies and edited the draft protocol. MHC provided input to the review conceptualization and edited the draft protocol. DNJ helped conceive the study and edited the draft protocol. SES helped conceive and design the study, and edited the draft protocol. All authors read and approved the final protocol.JMH is a practicing internist, geriatrician and clinical pharmacologist, and trainee with the University of Toronto Eliot Phillipson Clinician Scientist Training Program and Li Ka Shing Knowledge Institute. ACT is a systematic review methodologist and co-directs a knowledge synthesis center with SES. DNJ is a practicing internist and clinical pharmacologist, and Division Head of Clinical Pharmacology at the University of Toronto. SES is a practicing internist and geriatrician, and Division Head of Geriatric Medicine at the University of Toronto. MC is a biostatistician with the Li Ka Shing Knowledge Institute. LP is an information specialist with the Li Ka Shing Knowledge Institute and the University of Toronto.


Distatrix Mason from Central and South America, D. loretta, D. xanadon, D. vigilis, D. pitillaensis, D. pandora Grinter, n. sp., and D. antirrheae Whitfield & Grinter, n. sp., are described from large-scale caterpillar inventory endeavors, mostly from the larvae of geometrid moths. Biological information and diagnostic features that distinguish these species from other previously described Distatrix, especially those from the Neotropical region, are provided; and the first key to New World species is presented. The new discoveries expand our knowledge of the World's widespread Distatrix fauna by about a third, suggesting that similar survey efforts in other poorly sampled regions will reveal numerous additional undescribed species.Six new species of the genus  Distatrix Mason have recently been reared from biological inventories in the Neotropics. This unusual genus of braconid wasps (Microgastrinae) has often been confused with the superficially similar but much more common and diverse genus Glyptapanteles Ashmead, resulting in few reliable records of its distribution and biology. Mason  that the genus might be related to another of his newly erected genera, Rasivalva Mason. More recent combined analyses of molecular and morphological data  (Noctuidae) on Missouri Ozark oaks. Thirteen other species have been described worldwide  . Each of these species comes from a distinct ecosystem, ranging from 285 meters (D. vigilis) to 1220 meters (D. xanadon), and has a specific and different caterpillar host species. Since the establishment of the ACG inventory there have only been seven specimens of Distatrix reared from approximately 390,000 wild-caught caterpillars (Janzen pers. comm.). A leg from each of these specimens has been DNA barcoded by the BOLD initiative (http://www.barcodinglife.org) (labeled with DHJPAR#). Of the six specimens sampled only four have produced viable DNA . Distance based analysis of the sequences show substantial nucleotide differences , that in conjunction with diagnostic morphological and biological characters, establishes new species status (Four of the six new species were reared from specimens collected for the caterpillar and parasitoid inventory of the Area de Conservación Guanacaste (ACG), north-western Costa Rica (s status .www.caterpillars.org) and by related rearing projects in Panama (Barro Colorado Island) and Costa Rica , Distatrix antirrheae has only once been reared from eastern Ecuador. This second species of Distatrix is known from the rarely collected Morphinae butterfly, Antirrhea adoptiva porphyrosticta . This host preference, along with several morphological characters, strongly corresponds to many of the eleven Old World Distatrix and is unique amongst all known New World taxa.The remaining two new species were reared from the Caterpillars and Parasitoids of the Ecuadorian Andes project ( Station ). While Distatrix pandora were collected in larger numbers due in part to a concentrated search for the host caterpillar Eois (Geometridae). Because of the small and cryptic nature of the Eois caterpillar (Eois caterpillars (72 species), and reared out over 400 parasitic Hymenoptera and Diptera. This focused collection of Eois yielded higher diversity of parasitoids than the assemblage reared from Eois in Panama and Costa Rica  and included multiple species of Braconidae , Ichneumonidae (27%), Tachinidae (22%), Pteromalidae (2%), and Eulophidae (1%). If D. pandora is specialized on Eois then it is surprising that this rich yield of parasitoids did not include more specimens of Distatrix. Nevertheless, based on the high levels of Eois parasitism and the high densities of Eois , the Ecuador site will certainly prove to be an important location for future work on Distatrix-Eois relationships and the importance of Distatrix pandora in geometrid parasitoid communities.Specimens of erpillar  special Distatrix parasitize a wide range of hosts: Papilionidae, Nymphalidae, Noctuidae and Arctiidae  and D. solanae Whitfield the behavior of producing stalked cocoons on nearby substrate of the host.Below we provide several biological details and morphological notes that will help distinguish all known Neotropical species of ws Mason  for bodyws Mason  for wingby Mason  possesseby Mason . Five ofHolotype female. — Body length 2.65mm; forewing length 2.96mm.Head: black except yellow-orange palpi, honey-orange labrum, mandibles, clypeus and frontoclypeal suture. Scape mottled black and brown, flagellomeres dark brown. Frons barrel-shaped and medially taller than broad due to size of eyes, weakly punctate. Eyes extremely large, covering most of lateral view of head. Interior margin of eyes narrowing slightly towards clypeus then tapering off ventrally. Antennae longer than forewing length and tapering gradually towards apex. Ocelli in an equilateral triangle, distance to eye-margin from lateral ocellus equal to one of its longer diameters. All but distal flagellomeres with two ranks of placodes.Mesosoma: . The caterpillar was described as “green with two white lines down the back, with dots on them”. Cocoons formed on 18 August 2005, the wasps eclosed on 22 August 2005. Two Distatrix escaped during specimen transfer and two died and rotted after eclosion (Five wasp larvae emerged from the single undetermined Geometridae caterpillar 98-SRNP-7305 found as a penultimate instar feeding on eclosion .loretta to honor his mother Loretta Grinter.The senior author chose the name Distatrix pandora in morphology and resembles Distatrix xanadon and D. antirrheae in having a higher proportion of black coloration. It has the same enlarged eyes as displayed by D. belliger (Wilkinson) and D. solanae Whitfield . With D. teapae Nixon and at least eight other Distatrix species, D. maia Nixon, D. formosus (Wesmael) and D. solanae Whitfield, and all herein described new species (but not D. belliger), it shares a modified distal front tarsomere which is excavated apically on ventral side and bears a strongly curved, modified, spine.This species is similar to pandora or teapae. As with most other Neotropical species of this genus, the eyes of at least the females are enlarged, possibly suggesting a nocturnal or crepuscular host searching period; moderate xanthic coloration also suggests the latter. This species also shares the trait of gregariousness (having more than one parasitoid per host caterpillar) with D. antirrheae and these Old World species: D. cerales Nixon, D. geometrivorus (de Saeger), D. anthedon Nixon, D. gratiosus (Wilkinson) and D. pallidocinctus (Gahan); this is the only known species of gregarious Distatrix to spin pedunculate cocoons (other gregarious species spin loose cocoon masses or the cocoons stand up among host hairs) (Mason 1965).The raised medial area of metasomal tergum II is defined by divergent groves that fade posteriorly. This, and a lighter and reduced yellow coloration, will differentiate this species from Holotype female. Body length 3.09mm; forewing length 3.27mm.Head: dark orange except for light yellow palpi, labrum and mandibles; interior margins of scape black, margins around ocelli darkened, black flagellomeres. Frons broad, barrel-shaped, slightly diverging both dorsally and ventrally due to shape of eyes, only slightly taller than broad, forming almost a square. Eyes of normal microgastrine size. Antennae slightly longer than forewing length, all but distal flagellomere with two ranks of placodes. Lateral ocellus separated from eye-margin by a distance equal to 1.5x its longer diameter.Mesosoma: . Caterpillar found as a second instar larva with “dark coffee color and coffee color head” (http://janzen.sas.upenn.edu). Cocoons formed on 11 June 2006 and eclosion on 25 June 2007.Reared from a single undetermined geometrid feeding on xanadon is derived from Orson Welles's Xanadu; due to the unusual construction of the cocoon and high elevation from which it is known.The name Distatrix is substantially different from other known Neotropical species and has normal microgastrine eye size in common with D. antirrheae and all eleven species of Old World described Distatrix. This species otherwise appears similar in morphology to D. loretta as described above. The overall black coloration, extremely arched profiles of propodeum and tergite I and larger setae along the ovipositor sheaths make this Distatrix unmistakable in comparison with other known Neotropical species. The normal microgastrine eye size suggests a closer relationship with Old World species of Distatrix or quite simply a diurnal host searching period. The superficial resemblance to D. antirrheae may also serve to predict the cocoon type  for D. xanadon.This species of Holotype female. Body length 2.91mm; forewing length 3.22mm.Head: deep brown-black except for honey-orange mandibles, clypeus, scape and pedicel. Palpi pale yellow to whitish. Flagellomeres honey-brown and darkening apically, all but distal flagellomeres with two ranks of placodes; antennae slightly longer than forewing length. Frons barrel-shaped due to size of eyes, dorsal crease notched apically. Eyes extremely large, forming most of visible portions of head in lateral view. Lateral ocellus separated from eye margin by extremely slim strip, nearly touching.Mesosoma: , found feeding on Celtis trinervia Lam. (Celtidaceae) as a pen-pen-penultimate instar. Cocoons formed on 20 June 2004 and eclosion on 23 June 2004.Reared from a single Noctuidae, This species is named for its unusually large eye size.Distatrix teapae Nixon, sharing with it and D. belliger (Wilkinson), D. loretta, xanadon, pitillaensis, pandora, the enlarged eyes . With D. teapae and at least all other described species herein and D. maia (Nixon) and D. formosus (Wesmael) (but not D. belliger), it shares a modified distal front tarsomere, which is excavated apically on the ventral surface and bears a strongly curved, modified, spine.This species closely resembles D. teapae Nixon, D. pitillaensis and D. pandora most notably in wing venation, coloration, propodeal morphology and placement of antennae on the head. In the new species the wing venation is reduced in both structure and coloration with the absence of 2Cua and 3RSb and nearly white regions of M+CU, 1-1A, C+SC+R and R1 , found feeding on Siparuna thecophora Poepp. & Endl (Monimiaceae) on 22 May 2004, cocoon formed on 28 May 2004 and eclosed on 6 June 2004. Male reared from a single indeterminate Geometridae “same as 98-SRNP-11500” feeding as pen-penultimate instar on Croton morifolius (Euphorbiaceae) on 23 July 1998.Female reared from a single The name of this species is derived from the type locality in Costa Rica.Distatrix teapae in coloration, sharing with it and D. belliger (Wilkinson), D. loretta, xanadon, vigilis and pandora the enlarged eyes . With D. teapae and at least the other species described herein, and excluding D. belliger, it shares a modified distal front tarsomere, which is excavated apically on the ventral side and bears a strongly curved, modified, spine.This species most closely resembles D. teapae most strikingly in the hind coxae of the female, showing a vastly reduced length that barely extends beyond tergite I . D. pitillaensis is also darker in coloration with a more dark honey-orange mesosoma and metasoma with dark brown margins.This new species differs from Holotype female. — Body length 2.62mm; forewing length 2.90mm.Head: . 1 male, ECUADOR: Napo Province, Yanayacu Biological Station and Center for Creative Studies, 5 km W. Cosanga, 2100m, 0°42′01.33″S, 77°44′00.00″W, 16 March 2002. 1 male, similar data except 3 June 2001.Holotype female: PANAMA: Barro Colorado Island. 9°09′N, 90°51-W, artificial island made up of 15km2 of lowland moist forest located in the Panama Canal (Gatun Lake), 2003. Paratypes: 9 females, 10 males, similar data except emergence and pupation dates. 9 females, 10 males, similar data except 2004. 1 female, similar data except 11 June 2001. 1 male, 1 female, similar data except 22 July 2003. 3 females, 3 males, similar data except 23 July 2003. 1 female, similar data except 25 June 2005. . 3 females, 7 males: COSTA RICA: Heredia Province. La Selva Biological Reserve, located at 100m on the Caribbean slope, 10°26′N 83°59′W  in Ecuador at 2100m, indicating a surprisingly diverse and large range extension.Eois nympha (Geometridae) feeding on Piper cenocladum C. DC. (Piperaceae). Hosts reared from Panama are from an undetermined species of Eois feeding on Piper aequale Vahl and Piper scheideanum Steud.. Two male specimens from Ecuador reared from an undetermined Geometridae.Single holotype female were reared from pandora because of the confusion between this and a similar species of Diolcogaster for which it may be mistaken.This species is named Distatrix teapae (Nixon), both in morphology and coloration, diverging only with the size of the lateral metapleural pit and angle of the lines on metasomal tergum II; sharing with it and D. solanae Whitfield, D. xanadon, pitillaensis and D. belliger (Wilkinson) the enlarged eyes . With D. teapae and at least D. maia (Nixon), D. formosus (Wesmael), D. loretta, xanadon, vigilis, pitillaensis, antirrheae (but not D. belliger), it shares a modified distal front tarsomere, which is excavated apically on the ventral side and bears a strongly curved modified spine.This species is almost identical to D. teapae in having an overall smaller body size, lighter orange coloration, and these slight morphological differences: Metasomal tergum II with anterior medial area defined by grooves diverging at an angle roughly 90°, posteriorly converging and forming a weak diamond shape, whereas in D. teapae the angle is greater than 120° , 00° 35.949S 77°53.403W, 2100m, rearing number 26548. Paratypes: 34 other females and 2 males, same data. Holotype and 20 paratypes deposited in U. S. National Museum, Washington, DC; other paratypes in Illinois Natural History Survey, Champaign, IL.Antirrhea adoptiva in Colombia; and quite possibly in other species throughout the genus Antirrhea for which the biology is poorly known .Known only from type locality. Parasitoid may be present in populations of the nominate subspecies of Antirrhea adoptiva porphyrosticta (Watkins), a rarely collected nymphalid feeding on Chusquea scandens Kunth (Poaceae). For additional details on this host and its biology, see Greeney et al. (this issue). In addition to the holotype and paratypes mentioned above, approximately 10 other adult wasps failed to eclose from the cocoon mass, so the brood size appears to have been around 45.The gregarious brood of parasitoids emerged from a caterpillar of The species is named after the genus of its host caterpillar.This species can be easily distinguished from its other known congeners in the New World by its combination of a whitish and posteriorly evenly narrowing first metasomal tergite and otherwise quite dark body, along with the normal-sized eyes in both sexes and the suggestion of a forewing areolet. The gregarious habit, host family and unstalked, wooly white cocoons are also distinctive among New World species, but would not be out of place among some Old World species.Paper copies of this article will be deposited in the following libraries. Senckenberg Library, Frankfurt Germany; National Museum of Natural History, Paris, France; Field Museum of Natural History, Chicago, Illinois USA; the University of Wisconsin, Madison, USA; the University of Arizona, Tucson, Arizona USA; Smithsonian Institution Libraries, Washington D.C. USA; The Linnean Society, London, England.