On page 7 of the PDF, the first two paragraphs of the Acknowledgments section should read as follows.Volume 3, no. 2, e00095-18, 2018, We acknowledge discussions with the Terence Hwa lab at the University of California, San Diego, where this work was initiated.This work was supported by the National Natural Science Foundation of China (grants 31700039 to M.Z. and 31700089 to X.D.), by startup funding from the Central China Normal University, and by U.S. National Institutes of Health grant R01GM095903 through Terence Hwa. M.Z. and X.D. especially thank the Institute of Science and Technology Development of the Central China Normal University for funding support.


Studies on malaria vectors in the Pantanal biome, Central Brazil, were conducted more than half a century ago.To update anopheline records and assess receptivity and vulnerability to malaria transmission.Five-day anopheline collections were conducted bimonthly in Salobra, Mato Grosso do Sul state, for one year. Indoors, mosquitoes were collected from their resting places, while in open fields, they were captured using protected human-baited and horse-baited traps near the house and at the Miranda River margin, respectively. Hourly biting activity outdoors was also assessed. Secondary data were collected on the arrival of tourists, economic projects, and malaria cases.Anopheles darlingi was the predominant species, followed by An. triannulatus s.l. Hourly variation in anopheline biting showed three main peaks occurring at sunset, around midnight, and at sunrise, the first and last being the most prominent. The highest density of all species was recorded near the river margin and during the transition period between the rainy and early dry seasons. This coincides with the time of main influx of outsider workers and tourists, whose activities mostly occur in the open fields and frequently start before sunrise and last until sunset. Some of these individuals originate from neighbouring malaria-endemic countries and states, and are likely responsible for the recorded imported and introduced malaria cases.A total of 24,894 anophelines belonging to 13 species were caught. The main Brazilian malaria vector Pantanal is a malaria-prone area in Brazil. Surveillance and anopheline control measures must be applied to avoid malaria re-emergence in the region. The main malaria-endemic area in Brazil lies in the Amazon Region; this region accounts for 99% of Brazil's  annual malaria cases , b. Howehttp://www.ibge.gov.br/home/), the Migratory Efficiency Index of MS in 2009 was 0.0712, which is considered high migratory turnover. The main economic activities of MS are agriculture (including livestock farming) and tourism. The tourist attractions in the Pantanal area are seasonal and bring in high revenues.The influx of malaria-infected people from malaria-endemic regions is a key contributing factor to MS and Pantanal's vulnerability. According to the IBGE  (t biome) .2, with extensively flooded areas and lowheight shrubs characteristic of Cerrado (Savannah-like vegetation) subject to well-defined periods of flooding and drought. Ecotourism, fishing, hunting, and free-range cattle breeding attract tourists from other parts of Brazil and abroad. There is also a great influx of people into Pantanal for work .The Pantanal biome is a vast flat marshland of 150,355 kmsalobra means ‘brackish’ in Portuguese and refers to the salinity of the river water . Approximately 37% of Miranda's population lives in rural areas (IBGE - Available from: http://www.ibge.gov.br/home/). The human population of Salobra is mainly concentrated on the banks of the Miranda River , a village situated at the margin of the Miranda River, in the municipality of Miranda, MS, Brazil . The coler water . The muner water . Mirandada River . The Panda River . In Pantda River  and the da River . The dryda River . RecordsAnopheline collections - Mosquitoes were collected in September and November 2010, and January, March, May, and July 2011 , and larval chaetotaxy. Some of the mosquito samples transferred live to the laboratory were used to obtain offspring for confirmation of species identification using morphological characters .undation   from September 2010 to July 2011 ; among tAnopheles deaneorum Rosa-Freitas and An. oryzalimnetes Wilkerson and Motoki. Regarding the Oswaldoi complex, only An. konderi s.l. Galvão and Damasceno could be confirmed to occur in the area. As expected, the species comprising the Triannulatus complex were detected: An. triannulatus s.s. (Neiva & Pinto), An. halophylusAn. triannulatus C An. arthuri Unti and An. galvaoi. Since only a few mosquitoes could be analysed by molecular methods, the results concerning species belonging to these complexes are hereafter referred to as An. albitarsis s.l., An. oswaldoi s.l. + An. konderi s.l., An. triannulatus s.l., and An. galvaoi + An. arthuri (http://cculi.fiocruz.br/).Morphological and molecular identifications revealed the presence of 13 species or species complexes in Salobra –III. Mol arthuri –IV. VoucAn. darlingi Root and An. triannulatus s.l., which together constituted 82.9% of the total specimens captured in Salobra. Species captures at lower frequencies were An. albitarsis s.l. (4.5%), An. oswaldoi s.l. + An. konderi s.l. (2.1%), An. mattogrossensis Lutz and Neiva (1.4%), An. galvaoi + An. arthuri (0.7%), and An. rondoni (Neiva & Pinto) (0.6%).The most frequent species or species complexes were An. darlingi was the predominant species from the late rainy to early dry season, while An. triannulatus s.l. was the prevalent anopheline from the late dry season to the beginning of the rainy period , while the most frequent species at peridomicile were An. darlingi and An. triannulatus s.l. .Among the 12,283 anophelines caught in the sunset collections, the largest portion was caught extradomicile , 60% of which were . at 28% . Only 0.An. darlingi, An. triannulatus s.l., and An. albitarsis s.l. - displayed some differences in their peaks of hematophagic activity among species as well as according to the season. For some anopheline species, the number of specimens captured did not exceed 25, making it difficult to assess their peaks of hematophagic activity.A total of 10,686 anophelines was caught in the three 18-h collections performed to characterise the biting peak in March, May, and July 2011. Hourly variation in anopheline collections essentially showed three main peaks of hematophagy, occurring at sunset, around midnight, and at sunrise . HoweverAn. darlingi and An. triannulatus s.l. in these two samplings varied from 38 to 40% and 33 to 60.2%, respectively.Ovaries of a total of 309 anophelines collected extradomicile were dissected in May 2011  and July 2011 . The proportions of parous females of Table I). In 2010, almost 70,000 tourists visited Brazil, with MS being the entryway (Table II), 39 of which were autochthonous or introduced cases in or near Miranda, the municipality where Salobra is located; 433 were imported malaria cases   , b. Most52 years .An. darlingi, as well as species belonging to the Albitarsis, Triannulatus, and Oswaldoi complexes , the landscape and the dry/rain regimen result in flooding of vast lowlands at the riverbanks, creating abundant and diverse breeding sites for mosquitoes, especially anophelines. This condition favours the diversity of anopheline fauna, as revealed by the detection of 13 species in this study; among these, An. darlingi. Coincidently, the number of tourists/visitors and outsider workers attracted by temporary farming and other activities peaks in the region in this period  in the transition between the rainy and dry seasons (May) as well in the early dry season (July). This biting pattern was previously observed elsewhere (All anopheline species, most markedly lsewhere . Notablylsewhere . This palsewhere .An. darlingi, considered the main malaria vector in Brazil, other anophelines such as An. triannulatus s.l. and An. albitarsis s.l. were observed at relatively high frequencies in Salobra. These species are considered putative secondary vectors and can sustain malaria transmission, especially when present at high densities (Besides ensities .In conclusion, in Pantanal, a malaria-prone area in Brazil, the high density of anophelines that are competent malaria vectors combined with the seasonal influx and movement of people increase the possibility of malaria-infected individuals moving from malaria-endemic regions to this receptive area and reinforce the vulnerability of this large area to malaria transmission. Continuous epidemiological surveillance should be carried out to prevent the recurrence of malaria outbreaks and the reestablishment of endemic malaria in Pantanal.


Chaeropus ecaudatus, is one of the most enigmatic Australian marsupials, which went extinct in the late 1950s probably as a result of European colonization. It is unusual in being the only marsupial to have evolved reduction of digits on both fore and hind feet, with the forefeet being pig-like (two toes) and the hind feet being horse-like (one toe). According to molecular phylogenetic analyses, Chaeropus diverged from other bandicoots (Peramelidae), and the bilbies (Thylacomyidae) by the mid-Late Oligocene. This is considerably earlier than suggested by the fossil record, with the current oldest specimens being Late Pleistocene in age. Here, I report the oldest fossils of Chaeropus, representing a new species, Chaeropus baynesi from the Late Pliocene to Early Pleistocene (2.47–2.92 Ma) Fisherman's Cliff Local Fauna, Moorna Formation, New South Wales, Australia, and extending the fossil record of the genus and family by at least 2 million years. Chaeropus baynesi is less high crowned than C. ecaudatus and lacks lateral blade development on lower molars, suggesting that it was unlikely to be grazing. This suggests that Chaeropus must have adapted rapidly to the drying conditions and changes in environments, and would have become a grazer in a very short period of time.The pig-footed bandicoot,  Left m2 ; left worn m2 Type locality and age. Fisherman's Cliff Local Fauna, Moorna Formation, New South Wales, Australia. It is Late Pliocene to Early Pleistocene in age (2.47–2.92 Ma).Diagnosis. The metaconule of the M3 of C. baynesi is less distinct as a cusp than that of C. ecaudatus, but the postmetaconule crista ends more posteriorly. The cristid obliqua on the m2 of C. baynesi differs from that of C. ecaudatus in being less developed , and ending in the centre of the tooth, instead of the lingual margin of the tooth. Overall, the teeth of C. baynesi are smaller in size and less high crowned than C. ecaudatus.Etymology. Named in honour of Dr Alexander Baynes for his contribution to the fossil record of Chaeropus.Description. The M3 is wider than it is long and distinctively high crowned, but less so compared with C. ecaudatus . StA is worn on the tooth. Both the parastylar and metastylar shelves are short but wide. The postparacrista ends at the base of an oval shaped StB. StD is also oval in shape and connects to the premetacrista lingually. A small StE is present posterior to StD, but no stylar crest is present. The protocone is low on the crown. The preprotocrista ends at the anterolingual base of the paracone. The postprotocrista joins the premetaconule crista posterobuccally. A distinct crest runs buccally from the junction of the postprotocrista and premetaconule crista on the talon between the paracone and metacone, and may be the continuation of the postprotocrista. The metaconule is not distinct and the postmetaconule crista continues to descend the talon posteriorly to end at the lingual base of the metacone. Tooth dimensions are 2.41 mm in length and 3.08 mm in width for NMV P38490.Chaeropus, but unlike Isoodon, the enamel does not cover part of the root. The anterior cingulum is short but wide. The trigonid is short, with the paraconid–metaconid being shorter than the protoconid–paraconid and protoconid–metaconid distances. The talonid is wider than the trigonid, with the hypoconid posterobuccal to the protoconid. Cristid obliqua is poorly developed compared with C. ecaudatus, and ends in the centre of the tooth. The posthypocristid is straight and ends at the posterior most corner of the tooth, where the small hypoconulid is located. The entoconid is large, almost triangular in shape, with buccally orientated preentocristid. Tooth dimensions are 2.71 mm in length, 1.69 mm in trigonid width, and 1.89 mm in talonid width for NMV P38492, and 2.75 mm in length, 1.72 mm in trigonid width, and 2.04 mm in talonid width for NMV P38496.The m2 is short but wide. The crown is higher on the buccal side than the lingual side, as is typical of 3.C. baynesi described here shows some of the typical synapomorphies for Chaeropus [. The upper and lower molars are high crowned, though less so that in C. ecaudatus. The upper molar lacks anterior and posterior cingula as in C. ecaudatus but this feature is also found in Perameles. In the lower molars of C. baynesi, high crown is achieved by the increase in height of the buccal cuspids, as in C. ecaudatus, unlike Isoodon or Macrotis, which achieve high crown by having enamelled roots. Another Chaeropus synapomorphy found in C. baynesi is the triangular/tear-shaped entoconid , which is conical in Perameles, Isoodon and Macrotis, and oval in New Guinea bandicoots . The only synapomorphy that C. baynesi is lacking is the development of the cristid obliqua which is lower and ends further buccal.The material of haeropus . The uppC. baynesi are also slightly smaller than C. ecaudatus, suggesting that it was a smaller animal. For example, the length and width of the M3 ranges between 2.8–3.31 mm and 3.82–4.44 m, respectively, in C. ecaudatus , significantly larger than the 2.41 mm in length and 3.08 mm in width of C. baynesi. The same is true for the m2 of C. ecaudatus  which is also larger than the m2 of C. baynesi . The exact body mass of C. ecaudatus is unknown, although it has been estimated at between 200 g [Chaeropus baynesi is likely to have been below 200 g considering it has smaller teeth than C.ecaudatus and therefore a smaller body mass.The teeth of en 200 g  and 300 C. ecaudatus have been conflicting, some anecdotes reporting flesh [C. ecaudatus may have been the smallest mammalian grazer to have ever existed. Chaeropus baynesi, however, is unlikely to have been a grazer, but more likely to be a mix-feeder or even omnivorous. Not only it is smaller than C. ecaudatus, it also has shorter crowns, suggesting that it would wear its teeth down faster under the same diet, and therefore has a shorter life. The poor development of the cristid obliqua on lower molars of C. baynesi provides one less blade for shearing of food, one of the important features found in C. ecaudatus to be able to reduce grasses to fine particles [Reports of the diet of ng flesh  or insecng flesh , and othng flesh ,17. Thoung flesh . Small-bng flesh , or evenng flesh . With itarticles .Chaeropus baynesi is a member of the Fisherman's Cliff Local Fauna in the Moorna Formation, Late Pliocene to Early Pleistocene in age [C. ecaudatus is also known from [C. baynesi is especially valuable for the understanding of the evolution of this group. With only 2 million years separating C. baynesi and C. ecaudatus, it is clear that evolution of herbivory in this group was quite rapid, and probably linked climate change during the Pleistocene, though, it is quite extraordinary for such a small mammal to have become a grazer.e in age . The loce in age . This isown from , suggestown from . While town from ,13. With


Moschus. Here we present a comprehensive description of cranial material, including the ear region and the dentition, of fossil moschid material from the Middle Miocene locality Steinheim am Albuch . This study provides the first exhaustive dataset for the cranial osteology of Micromeryx flourensianus, the most likely oldest true moschid. It furthermore reveals the presence of a second, so far undescribed moschid species, we here name Micromeryx? eiselei, in the abundant material from the locality. The two taxa can be clearly distinguished by characters of the skull, the ear region, the dentition, as well as by size. This evidences the sympatric occurrence of two moschid species in the locality Steinheim am Albuch.Moschids are enigmatic pecoran ruminants whose phylogeny is still not fully understood. So far we know only little of the family’s early evolutionary history and the origin of the modern genus,  Moschus Linnaeus, 1758. Moschidae are small pecoran ruminants with elongated upper canines in males lacking any cranial appendages. Moschus remains the only surviving genus of the family. It is restricted to montane woodlands in Asia 50], for Nomenclatural actshttp://zoobank.org/". The LSID for this publication is: urn:lsid:zoobank.org:pub:86EDF102-759B-4A77-83C7-7FAE99074C5A. The electronic edition of this work was published in a journal with an ISSN, and has been archived and is available from the following digital repositories: PubMed Central, LOCKSS.The electronic edition of this article conforms to the requirements of the amended International Code of Zoological Nomenclature, and hence the new names contained herein are available under that Code from the electronic edition of this article. This published work and the nomenclatural acts it contains have been registered in ZooBank, the 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 "MAMMALIA Linnaeus, 1758CETARTIODACTYLA Montgelard, Catzeflis and Douzery, 1997RUMINANTIA Scopoli, 1777MOSCHIDAE Gray, 1821Micromeryx Lartet, 1851Genus Micromeryx flourensianus Lartet, 1851Type species: Holotype: hitherto not determined  MNHN Sa 2957); material from Sansan  under revision; partly figured in Filhol ; stored at MNHN.NMB Sth.834 (compressed skull of skeleton); Sth.804 (fragments of upper and lower jaw of skeleton), NMB Sth.811 and Sth.812 ; NMB Sth.828a, NMB Sth.865, NMB Sth.866, SMNS 40500–1 , SMNS 46123 , SMNS 15776 , SMNS 12966 (fragmented mandibula with worn and fragmented post canine toothrow and incisor fragments), SMNS 46116 (right lower jaw fragment), SMNS 40252 , SMNS 42920 , SMNS 43062 , SMNS 46083 (maxilla fragment).NMB Sth.834 and NMB Sth.865, NMB Sth.866, SMNS 40500–1, SMNS 46123 comprise adult specimens. NMB Sth.828a and NMB Sth.811 were juveniles or at young adults. The more complete skull NMB Sth.834 was enclosed obliquely and was strongly fractured and fragmented.Micromeryx flourensianus from Steinheim a. A. possessed an elongated and low skull, with a broadened cranium and a facial part distinctly narrowing to anterior, similar to the modern genus Moschus. As the best preserved skull (NMB Sth.834) is strongly compressed estimations on the skull size are enhanced, but it appears to have been only slightly smaller than the modern Moschus moschiferus. A roughly rounded shape can be reconstructed for the orbita (observable in NMB Sth.834 and NMB Sth.811). Its relative size (in ratio to skull size) appears a little larger than in the modern Moschus.Generally Occipital Figs  and 4. The partes laterales and the squama occipitalis of the occipital can be observed in NMB Sth.834, where they are obliquely compressed. A narrow, but distinct crista nuchae  appearsx daamsi . The areC. elaphus.The pars basilaris shows a smooth surface, except of the two concave surfaces for the muscle insertion with the distinct posteromedial tuberculi musculares and the posterior protuberantia at the suture to the partes laterales . We coulSphenoid Figs  and 5. In NMB Sth.811 the ala ossis presphenoidalis forms the lower part of the medial wall of the orbita and meets the frontal here in an s-shaped suture line with the frontal reaching more ventral anteriorly. As in Moschus the ala ossis presphenoidalis is dorsoventrally higher than in C. elaphus, where the palatine more strongly intrudes into the medial wall of the orbita in dorsal direction. A relatively large foramen opticum  is located posterior to two smaller foramina. The anteroposteriorly oriented foramen orbitorundum is observable in NMB Sth.811 . The lamina vaginalis encloses the processus styloideus at least proximally. Thus the thympanohyal vagina is not fully visible from lateral view. However, as the lamina is fragmented the full extent cannot be reconstructed. The processus styloideus is separated from the processus jugularis of the occipital, comparable to the situation in Moschus. In C. elaphus this feature was variable in the studied specimens. There appears to be no contact between the tympanic bulla and the occipital in M. flourensianus from Steinheim a. A.x daamsi . The bulPars petrosa and bony labyrinth Figs  and 8. The petrosal and bony labyrinth of M. flourensianus from Steinheim a. A. were both described in detail by M. floure unclear ,53). M. ocristid . M. sorihe genus . M. flouonoideus .M. flourensianus from Steinheim a. A. and M.? eiselei in the cranium, mandibula, dentition, and especially in the ear region are too large to ascribe them to intraspecific variability. Both taxa are furthermore represented by both sexes and different ontogenetic stages. These differences are sufficient to clearly distinguish between the two species.In summary the morphologic differences between The main differences are:M.? eiselei but absent in M. flourensianus from Steinheim a. A.crest on the pars basilaris of the occipital present in M.? eiselei than in M. flourensianus from Steinheim a. A.processus jugularis more slender in M.? eiselei than in M. flourensianus from Steinheim a. A.tympanic bulla less rounded in M.? eiselei than in M. flourensianus from Steinheim a. A.petrosal less wide in ventromedial-dorsolateral direction in M.? eiselei than in M. flourensianus from Steinheim a. A.posterior bulla articulation in petrosal larger in M.? eiselei than in M. flourensianus from Steinheim a. A.anterior process of the tegmen tympani more delicate in M.? eiselei than in M. flourensianus from Steinheim a. A.tegmen tympani dorsomedially-ventrolaterally wider in M.? eiselei, divergent in M. flourensianus from Steinheim a. A.course of the vestibular aqueduct with respect to the common crus parallel in M.? eiselei than in M. flourensianus from Steinheim a. A.vestibular aqueduct longer in M. flourensianus from Steinheim a. A. in contrast to straight long endolymphatic sac in M.? eiseleishort pouch shaped endolymphatic sac triangular in M.? eiselei than in M. flourensianus from Steinheim a. A.general tooth size larger in M.? eiselei than in M. flourensianus from Steinheim a. A.posterior part of the upper P3 slightly more elongated in M.? eiselei than in M. flourensianus from Steinheim a. A.labial wall of upper P4 with stronger relief in M.? eiselei than in M. flourensianus from Steinheim a. A.labial wall of upper molars more pillar-like in M.? eiselei than in M. flourensianus from Steinheim a. A. (less distinct feature)p3 slightly more slender with a more open anterior valley and a less bulky mesolingual conid in M.? eiselei than in the ones of M. flourensianus from Steinheim a. A. (less distinct feature).metastylid and anterior cingulids stronger in the lower molars of Moschus in:Both taxa differ from the genus the arrangement of the foramina for the nervus hypoglossusthe course of the suture line between maxilla and palatinethe morphology of the petrosalthe position of the foramina mentaliaa less shortened premolar rowa simpler morphology in the p4the presence of an external postprotocristidless increase in tooth crown height.M.? eiselei and Moschus distinguishing both from M. flourensianus from Steinheim a. A. These are:There are however characters shared by the crest on pars basilaris of the occipitalthe parallel course of the vestibular aqueduct with respect to the common crusthe longer vestibular aqueductan elongated endolymphatic sac.Micromeryx? eiselei from Steinheim a. A. cannot be assigned to any known Micromeryx- or Hispanomeryx-species. It possesses a unique combination of characters in cranial, mandibular, and dental morphology: large size of teeth, the presence of a strongly pronounced external postprotocristid, a distinct lateral extent of the processus coronoideus, and a wider p4 than that observed in M. styriacus. Therefore, we consider it reasonable to erect a new species, although the postcranial material has not been analysed yet. M.? eiselei clearly differs from Moschus in the morphology of the cranium and the dentition. Interestingly, the characters shared by M.? eiselei and Moschus are only characters of the ear region and the basicranium, which are considered to be less plastic than e.g., dental data. However, considering the evolution of Pecora, some characters of the ear region are plesiomorphic [M. flourensianus from Steinheim a. A. and M.? eiselei might prove less stable than it now appears. However, the revision of the type material of M. flourensianus as well as a full analyses of the postcranial and the ear regions of more fossil Moschidae is in progress, but not yet completed. Therefore, we can assign M.? eiselei to a new species but not to a new genus, and thus decided with reservations to leave it in the genus Micromeryx at present.omorphic , and thuMoschus. Dental and cranial characters as well as features of the ear region reveal the presence of at least two species at the locality.The small ruminant remains from the Middle Miocene locality Steinheim a. A. can be assigned to Miocene Moschidae closely related to the living genus Micromeryx flourensianus from Steinheim a. A., is similar to the type material of M. flourensianus from Sansan, but differs by the presence of an “evolved” dentition. The larger taxon, Micromeryx? eiselei clearly differs from all Miocene Moschidae and is therefore referred to a new species.The smaller of the two, Hispanomeryx due their size and described as sympatric with Micromeryx flourensianus by Aiglstorfer et al. [The two moschid taxa described here from Steinheim a. A. originate from the same layer in the stratigraphic column based on the biostratigraphy of the embedded gastropods. The fossil assemblage shows a high percentage of semi-articulated partial skeletons and is embedded in lacustrine sediments, it did not result from a fluvial accumulation and represents an autochthonous taphocoenosis. Thus, independent of the further taxonomic status, there are two sympatric moschid species recorded from Steinheim a. A. This confirms once more that the co-occurrence of two moschid species is rather a more common than an exceptional phenomenon for the Miocene. However, it is the first time that this observation is made with unambiguous remains for European material outside of Spain  fragmentary teeth were tentatively assigned to? r et al. ; with thM. flourensianus and the Central/Western European record of the family. Combining the data published here with the study of the type material, and postcranial remains, as well as with data for the ear region of more Miocene Moschidae will improve the knowledge on the early evolutionary history of the family and the origin of the recent genus Moschus.The exhaustive description of the morphology of the ear region combined with data on cranial, mandibular, and dental material represented here is the first step towards a better understanding of the possibly oldest moschid species, S1 Text(DOCX)Click here for additional data file.S1 File(NEX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.


Improving research practices is a community project, and as such we are excited to have received many responses to our paper . Overallcan still be published, right alongside pre-registered analyses. They are an important tool for generating new hypotheses, but they should be distinguished because they are, by definition, influenced by viewing the data. Lastly, researchers do not need to worry about protecting their ideas: preregistrations can be embargoed for up to 4 years on the Open Science Framework or as long as desired on AsPredicted.org. Besides, opening up preregistration (or part of it) right at conception could promote collaboration, mirroring the approach taken in epidemiology . As lonM.I. was funded by a grant from the Natural Environment Research Council  awarded to T. Burke. I.S.W. was funded by an ERC grant to B. Pujol (ANGI 681484).


Helicobacter pylori (H. pylori) is a bacterium infecting 28% to 84% of subjects, depending on the population tested. However, the implication of H. pylori in cardiac arrhythmia is poorly understood. We performed this meta-analysis with an aim to identify the association between arrhythmia and H. pylori. We searched PubMed, Embase, Web of Science, and the Cochrane library databases to select studies on the association between arrhythmia and H. pylori. In the arrhythmia group, 392 (58.1%) were H. pylori-positive and in the control group 640 (47.8%) were H. pylori-positive. Compared to the controls, the infection rate of H. pylori was higher in patients with arrhythmia than in controls (odds ratio (OR) = 1.797, 95% confidence interval (CI): 1.081–2.988, p < 0.05). Subgroup analysis indicated that H. pylori infection was a risk factor for atrial fibrillation in Asia and Africa. Therefore, a correlation between H. pylori infection and arrhythmia may exist and H. pylori eradication may decrease the occurrence of arrhythmia, especially in Asia and Africa.Arrhythmia is a common disease around the world and  Helicobacter pylori (H. pylori) infection has been noted to be associated in previous studies. Therefore, we aimed to analyze the relationship within H. pylori infection and arrhythmia.Arrhythmias are common and are increasing, particularly atrial fibrillation. Arrhythmias may be divided into different categories based on various criteria. For instance, based on the beating rate, arrhythmias include bradyarrhythmia and tachyarrhythmia. In addition, arrhythmias can be divided into supraventricular and ventricular arrhythmias depending on the pathogenic area . The lifH. pylori is a Gram-negative microaerophilic bacterium and has been identified as the main pathogen for gastritis, peptic ulcer, and gastric cancer [H. pylori is also found to be associated with arrhythmia. In 2005, Montenero et al. revealed for the first time a highly significant correlation between H. pylori infection and atrial fibrillation [H. pylori strains were related to idiopathic dysrhythmias [H. pylori infection [H. pylori is implicated in the pathogenesis of arrhythmia is inconclusive. Recent studies have suggested that some microorganisms’ infection could contribute to the development of arrhythmia, including atrial fibrillation, through the inflammation cascade . H. pyloc cancer ,6,7. Besillation . Other ghythmias . Howevernfection . Thus, wH. pylori. The findings strengthening the association between arrhythmia and H. pylori infection could provide a new approach for treatment of stubborn arrhythmia.We conducted this meta-analysis to obtain a more comprehensive estimate of the association between arrhythmia and The meta-analysis was carried out according to the guidelines of the PRISMA statement and MOOSE ,12. We sH. pylori infection and arrhythmia; (3) H. pylori infection was diagnosed through the urea breath test (UBT) and/or serology and/or histology and/or culture in both case and control groups; (4) the studies were published in English. Articles were excluded if they were any of the following: (1) review articles, meta-analyses, letters, commentaries, or abstracts presented in conferences; (2) duplicates or continued work of previous publications; (3) studies without complete data; (4) not in English.Two reviewers (Jin Yan and Qiang She) checked the abstracts of the studies and read the full-texts if necessary to identify the final included studies. When disagreement appeared, we discussed and consulted the third reviewer (Yifeng Zhang). In addition, we turned to the original authors for more detailed data if necessary. Eligible studies strictly met the following criteria: (1) the studies utilized a case-control, prospective, or cross-sectional study design; (2) the studies contained sufficient data for investigating the association between To ensure the quality of the meta-analysis, the Newcastle-Ottawa quality assessment scale (NOS) was used and each item in the NOS was assessed for all included studies. The final score ranged from zero to nine stars for each study and a score of five stars or more was regarded to be optional ,14. Two H. pylori infection detection, arrhythmia group with H. pylori (+/−), control group with H. pylori (+/−), clinical characteristics of the populations, and other substantial information.After quality assessments, all authors extracted information from each paper and organized the data in Excel  independently. The following data were retrieved from each study: the first author, the year of publication, the country and continent, study design, study size, type of arrhythmia, diagnosis method for H. pylori infection and arrhythmia. Heterogeneity was assessed by the test of inconsistency index (I2) and the Cochran-Q method. I2 value more than 50% or p value less than 0.1 suggested the presence of heterogeneity and the random effects model was applied. We also performed subgroup analyses to seek the sources of heterogeneity. In addition, sensitivity analysis was conducted to evaluate whether omitting one study in each turn substantially altered the main results. Finally, the publication bias was analyzed by the funnel plots . Statistical analysis was conducted utilizing Stata 12.0  software.In this meta-analysis, we calculated dichotomous variables using the odds ratio (OR) with the 95% confidence interval (CI) to measure the strength of the association of Helicobacter cinaedi with atrial arrhythmias, and in the other paper we failed to extract sufficient data [The review process is shown in ent data ,15. Thusent data ,18,19,20H. pylori infection, and the number of H. pylori positive and negative subjects in arrhythmia and control groups. Demographic characteristics including age and gender are shown in The eligible articles were published from 2006 to 2015 accumulating 675 arrhythmia patients and 1339 controls. All the included studies were conducted using a case-control study design. The studies’ characteristics are presented in H. pylori infection (58.1%), compared to 47.8% in the control group (640/1339). There was significant heterogeneity in the included studies . Therefore, we used the random-effects model. The obtained pooled OR was 1.80 (95% CI: 1.08–2.99) (p = 0.024). Thus, we conclude that there may be a relationship between H. pylori infection and risk of arrhythmia.In the seven eligible studies, 392/675 patients in the arrhythmia group were positive for 08–2.99)  and the H. pylori infection correlated with idiopathic dysrhythmias . In the other six studies investigating the relationship between H. pylori infection and atrial fibrillation, the pooled OR was 1.53 . We also found that the subgroup of the region played an important role in the heterogeneity of the results. Compared to those studies in America  and Europe , investigations in Asia  and Africa  revealed that H. pylori was significantly related to atrial fibrillation. In addition, there was no significant heterogeneity in separated sub-groups   . We furtp > 0.1) , indicatp value was 0.23. Thus, there was a publication bias in this meta-analysis.To investigate the potential publication bias present in this study, the included studies were evaluated using Begg’s test. As shown in H. pylori infection and arrhythmia with a total of 2014 subjects. Overall, the H. pylori infection rate in arrhythmia patients  was significantly higher than that in control individuals  , indicating that H. pylori may be related with arrhythmia. In these seven studies, six studies investigated the association between H. pylori infection and atrial fibrillation, a common type of arrhythmia. However, the result was not significant . Then we divided the studies according to their origin and found that H. pylori infection was significantly associated with atrial fibrillation in Asia and Africa, but not in Europe and North America. This result suggested that H. pylori infection might play a role in the development of atrial fibrillation in the Asian and African population. In this study, a total of seven case-control studies published between 2006 and 2015 that met our study criteria were included in this meta-analysis. We provided an overview of the relationship between H. pylori infection can induce arrhythmia via activating inflammatory process. H. pylori is a bacterium that colonizes in the gastric epithelium and has been identified as a group 1 carcinogen for gastric cancer [H. pylori infection, the correlation between H. pylori and arrhythmia has begun to emerge, but the conclusion remains under debate. For example, in Italy, Montenero et al. have shown that atrial fibrillation significantly correlates with H. pylori infection; however, Platonov et al. did not find such a correlation [H. pylori infection, inflammation, and arrhythmia, and they have found that tumor necrosis factor-α, interleukin-6 and C-reactive protein, which are markers reflecting the degree of inflammation, were closely related to the type and the duration of atrial fibrillation [H. pylori virulence factor cytotoxin-associated gene A showed ability to cross-react with the antigens of endothelial cell membranes [H. pylori is both an inflammatory agent and an immune regulator, with virulence factor vacuolating cytotoxin A (VacA) efficiently blocking proliferation of T cells by inducing a G1/S cell cycle arrest [H. pylori might modulate a T helper (Th) 1/Th2 balance and H. pylori VacA to induce Th1 suppression and might abrogate T regulatory cell functions, subsequently resulting in an increase in the risk of atrial fibrillation [Arrhythmia is a common disease that badly influences the quality of life . Researcc cancer . With threlation ,10. In oillation ,26. In aembranes . On the e arrest ,28,29. HH. pylori. In Asia and Africa, the H. pylori infection rate was higher than that in western countries, which could be related to the different standards of hygiene and socioeconomics [H. pylori infection in Asia and Africa, but not in developed countries in Europe or North America.We also found heterogeneity in our analysis. Among these seven studies, six studies were about atrial fibrillation and one study investigated idiopathic dysrhythmias. The result for atrial fibrillation was not significant. Then, we also applied subgroup analysis based on origin and found that the continent where the study was conducted also presents a source of heterogeneity. The results were consistent with the different infection statuses of conomics . TherefoH. pylori infection and therefore the control group should be socioeconomically matched to the arrhythmia group. Thirdly, when serological tests of H. pylori infection are applied, it was difficult to distinguish between past and active infection conditions. Fourthly, although we conducted an intense search in Pubmed, Embase, Web of Science, and the Cochrane Library, there might still be published studies in other databases that we missed. Also, we only reviewed papers written in English. A publication bias was found in the meta-analysis. Thus, a further systematic review extended to other languages should also be considered. In addition, some studies with negative results might not have been published, which could cause unavoidable publication bias in our study.Our meta-analysis also has some limitations. Firstly, all included studies were case-control studies and the quality of this type of study was not as high as that of randomized clinical trials. Secondly, the socioeconomic status of the subject has been identified as a high risk factor for H. pylori should be considered as one of the risk factors for atrial fibrillation in Asia and Africa and may also be related to idiopathic dysrhythmia. However, large-scale and prospective studies examining the precise role of H. pylori in the development of arrhythmia will still be needed to further corroborate our conclusions. Our findings suggest that in Asia and Africa, eradication of H. pylori should be considered as an appropriate therapeutic regimen for some intractable atrial fibrillation cases.In conclusion, our study demonstrates that


Carex (Cyperaceae). In four important sections of the northern regions , the frequent occurrence of hybrids often renders the identification of “pure” species and hybrids difficult. In this study we address the origins and taxonomic rank of two taxa of section Vesicariae: Carex rostrata var. borealis and C. stenolepis. The origin and taxonomic status of C. stenolepis has been the subject of substantial debate over the years, whereas C. rostrata var. borealis has received very little attention in the years since its first description in the 19th century. By performing an extensive sampling of relevant taxa from a broad distribution range, and analyzing data from fifteen microsatellite loci developed specifically for our study together with pollen stainability measures, we resolve the hybrid origins of C. rostrata var. borealis and C. stenolepis and provide new insights into this taxonomically challenging group of sedges. Our results are in accordance with previous findings suggesting that C. stenolepis is a hybrid between C. vesicaria and C. saxatilis. They are also in accordance with a previous proposition that C. rostrata var. borealis is a hybrid between C. rostrata and C. rotundata, and furthermore suggest that both hybrids are the result of multiple, recent  hybridization events. We found little evidence for successful sexual reproduction within C. rostrata var. borealis and C. stenolepis, but conclude that the common and recurrent, largely predictable occurrence of these taxa justifies accepting both hybrids as hybrid species with binomial names. There are, however, complications as to types and priority names, and we therefore choose to address these problems in a separate paper.Hybridization is frequent in the large and ecologically significant genus  Carex (Cyperaceae). As an example, among the more than one hundred Carex species present in Norway, approximately 60% partake in some degree of hybridization, producing 113 hybrid combinations C. rostraHylander  providesC. stenolepis samples included in the study of Ford et al. [C. stenolepis (incl. C. grahamii) consists of largely sterile hybrids and should not be regarded as a “good” species . Furthermore, considering the use of potential pollen stainability measures, they pointed out that “while this technique allows for the detection of obviously sterile pollen grains it cannot be considered a precise estimate of fertility since stained grains are not necessarily viable”, thereby regarding pollen stainability as a rough measure of pollen sterility rather than fertility. Ford et al. [Even though some of the d et al.  had quitd et al.  found thC. rostrata var. borealis and C. stenolepis can exhibit moderately high levels of pollen stainability, this does not necessarily indicate that the taxa are able to reproduce sexually. Sexual reproduction depends, in the final instance, not on production of pollen grains but on production of seeds. According to P.W. Ball in [C. stenolepis but was unable to find plants with well-formed and mature achenes. Hylander [C. stenolepis plants with seemingly empty perigynia, but argued that this was not necessarily a sign of hybridity, as failed fruit set could be due to external factors such as frost or flooding, and that fruit set frequently fails also in other species of section Vesicariae. An additional factor may be failing pollination due to weather conditions in the comparatively short period with male anthesis.While it is clear that individual plants of both  Ball in , B.A. FoHylander  also fouC. rostrata, C. rotundata, C. saxatilis, and C. vesicaria), the hybrid index results and the data on pollen stainability reported in both this study and the study of Ford et al. [C. rostrata var. borealis and C. stenolepis are sexually reproducing species. It is therefore very unlikely that many if any of the stands have undergone significant evolutionary changes after the hybridization events. The very low number of private alleles in the two hybrid taxa compared to the high number of alleles shared with their respective parents indicates that the observed stands are based on single hybridization events, with little subsequent introgression with their parents and little or no stabilization by independent sexual processes. The STRUCTURE analysis clearly showed that both C. rostrata var. borealis and C. stenolepis combine approximately equal amounts of genetic material from their parents; again signifying recently formed hybrids.Although we agree that fruit set often can fail even in the four primary species  ranged from a rather modest 17 to 154 years old, the age of two clones of C. ensifolia Turcz. ex Ledeb. subsp. arctisibirica Jurtz. (= C. bigelowii Torr. subsp. ensifolia (Turcz. ex V. Krecz.), on the authority of [Vesicariae are largely clonal and able to form widespread and dense mats perhaps consisting of only a single genet . This is true also for the hybrids C. rostrata var. borealis and C. stenolepis.The frequent and extended stands of both our hybrids are due to frequent co-occurrence of the parents, frequent hybridization, and extensive clonal growth of the hybrids. One explanation why r et al.  estimateority of ) was estC. rostrata var. borealis as an understorey in swards of C. rostrata, is a common situation, and C. stenolepis is rarely found within or close to stands of either C. saxatilis or C. vesicaria. However, it is important to note that the dispersal ability of such hybrids is very limited. The perhaps only possible diaspores are rhizome fragments, and the most likely agents of dispersal are water or birds.If the predominant mode of reproduction in these plants is asexual , their success does not rely much on production of viable pollen or successful fruit set. One might argue for the treatment of these taxa as hybrid species as they are widespread and form constant “populations”  over a long period of time, certainly centuries and probably millennia. Given the long period of time involved, the parental species may have disappeared whereas the hybrids have remained, especially as the mires change during paludification and acidification. It is unlikely that competition plays any significant role. Co-occurrences, e.g., with Carex rostrata var. borealis and C. stenolepis have been treated rather different taxonomically. Whereas the latter has been extensively discussed in numerous floras and other publications, the former has received almost no attention in the years since its description. Carex rostrata var. borealis has been, and still is, regarded merely as a variety of C. rostrata by the majority of authors, the exception being Elven [C. stenolepis has rather focused on whether it should be accepted as a sexually reproducing species  or  or Carexs . As theC. rostrata var. borealis and C. stenolepis are swarms of clonal hybrids that have been formed multiple times across various locations. Contrary to Hylander’s [C. stenolepis plants are (at least) vegetatively very similar to one another, we found that the morphology of C. stenolepis varied quite a lot between locations, more so than the morphology of C. rostrata var. borealis did (field observations by A.T.M. Pedersen and R. Elven). Nevertheless, it is fairly easy to correctly identify both hybrids and to distinguish them from their parents. Also, C. rostrata var. borealis and C. stenolepis are much more common than other hybrids in section Vesicariae, and unlike these, they are significant constituents of the Fennoscandian mire vegetation, with regular and to a large degree predictable occurrence within consistent ranges, preferring habitats overlapping with, but not identical to, those of any of the four parents , but not in the swamps that C. vesicaria prefers).The results from this study affirm that both lander’s  claim thC. rostrata var. borealis together with a morphological review of this hybrid, and a reassessment of the nomenclature of what currently passes as C. stenolepis, will be presented elsewhere .Cayouette and Catling  wrote: “S1 Fig(PDF)Click here for additional data file.S2 FigMarkers Carspe_6867s and Carspe_2310c were excluded from this analysis as they did not amplify in all samples.(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.


Taenia taeniaeformis (T. taeniaeformis). Cysticercus fasciolaris (C. fasciolaris), the larval stage of T. taeniaeformis, develops in small rodents which act as intermediate host. The aim of this study was to estimate the prevalence of C. fasciolaris in brown rats (Rattus norvegicus) in the densely human populated parishes, St. George's and St. David's of Grenada, West Indies. One hundred and seventy rats were trapped near the residential areas from May to July, 2017 and examined for C. fasciolaris in their liver. Of the 170 rats 115  were positive for the larval stage of T. taeniaeformis. One to three cysts were observed in each liver, containing a single larva in each cyst. The prevalence was 77.9% in St. George and 59.1% in St. David which is a significant difference (p < 0.05) between the two parishes under study. Based on gender, prevalence in males was 60.9% and females 74.7%. Significant difference was observed between young and adult rats (p = 0.03). Prevalence in young rats was 45.0% compared to adults (70.7%). Further study of risk assessment in the cat population in areas of the present research is strongly suggested.Cat is the definitive host of Taenia taeniaeformis is a cestode parasite found in the intestine of cats as final host. Wild rodents, mainly mice, various species of rats, and voles act as intermediate host for the parasite. The intermediate hosts get infected through ingestion of contaminated feed, water, and beddings from eggs of the parasite voided by cats. Eggs develop into larval form (metacestodes) in the liver of intermediate host. The larval form of T. taeniaeformis is called C. fasciolaris. Taenia crassicollis, Hydatigera fasciolaris, Strobilocercus, and bladder worm are synonyms of Cysticercus fasciolaris [ C. fasciolaris develops mainly in the liver of rodents and contains larval stages of the parasite. Occasionally cysts also develop in the abdominal wall and kidney, filled with purulent exudate without larvae [ T. taeniaeformis have been reported [ C. fasciolaris. Although rare, humans get infected with eggs of T. taeniaeformis from cats [ciolaris . C. fasct larvae . A smallreported –5. Cats rom cats .T. taeniaeformis has been reported in rodents and cats worldwide. The report of C fasciolaris particularly, in brown rats (R. norvegicus), is from India [ Angiostrongylus cantonensis (A. cantonensis) in lung/heart of R. norvegicus [ C. fasciolaris in the liver of (29.6%) rats were also reported. As far as authors are aware, there is no published report of C. fasciolaris in brown rats in other Caribbean nations. The aim of this report is to estimate the prevalence of C. fasciolaris in brown rats from Grenada and compare with the previous report.om India , 8, Koreom India , Malaysiom India , Serbia om India , and USAom India . In Grenrvegicus , lesionsThe project (Detection of zoonotic pathogens in brown rats in Grenada) was approved by the Institutional Animal Care and Use Committee (IACUC # 16009-R) of St. George's University Grenada.2. The country with low hills, small trees and shrubs, and tropical climate is most suitable for the existence of brown rats. The country is divided into six parishes. The parishes of St. George and St. David were selected for sampling because of their dense human population compared to the other four parishes.Grenada is the southernmost country in the Caribbean Sea with an area of 348.5 Km (R. norvegicus) belong to genus Rattus under the family Muridae [Brown rats or Norway rats Muridae . They ar Muridae . l × 15 cm w × 15 cm h) with cheese and or various local fruits as bait. Attempts were made to trap the rats near the residential buildings. Trapping in both parishes was conducted near 10-meter periphery of human dwellings. Traps were placed in the evening and visited next day during the morning. Traps with rats were covered with black cloth and transported to the necropsy laboratory of the school of veterinary medicine, St. George's University, Grenada, and transferred to the anesthesia machine. Rats were anesthetized using isoflurane in oxygen via anesthesia machine (portable vet anesthesia machine isoflurane vaporizer VET CE), manufacturer DRE .One hundred and seventy rats were collected live from 1st May to 14th July, 2017, using live traps .The data was analyzed by the statistical analysis: Fisher's exact test, using graphical statistical software  positivity. The results for the prevalence are included in  C. fasciolaris by parish was statistically significant (p < 0.05). Risk factors being similar in both parishes, this difference in prevalence is not well explained. Further research involving more number of rats is suggested to answer the difference. Previous researchers reported in brown rats a prevalence of 100% in the Philippines [ A. cantonensis in brown rats, researchers found 29.6% rats also infected with C. fasciolaris. Variations in the prevalence of C. fasciolaris in different countries indicate infection risk factors, including seasonal variation in the infection pressure on the intermediate hosts [Out of 170 brown rats examined, 115 showed lesions oflippines , 33.3% ilippines , and 29.te hosts . PrevaleOn gross examination of liver of infected rats, one to three cysts were found in each liver . Size of C. fasciolaris. The wall of cysts varied in thickness from thin connective tissue capsule in mature C. fasciolaris and thick wall of connective tissue in juvenile C. fasciolaris. These findings are consistent with Lee et al. [Histopathology of the liver showed minimal pathological lesion in the liver parenchyma, except in and around the cysts. The cysts had a central lumen which containede et al. . Similare et al.  we also  C. fasciolaris according to gender in the present study is included in  C. fasciolaris in 66.7% male and 89.5% female in St George's parish compared to 56.3% male and 62.2% female in St David's parish. In our study, there was no significant statistical difference between male and female. Lee et al. [ C. fasciolaris among male and female rats. However, contrary to our findings Rodríguez-Vivas et al. [The prevalence ofe et al.  also reps et al.  found hi C. fasciolaris in young and adult rats are tabulated in p < 0.05). Our observation is in accordance with previous researchers [The results for prevalence ofearchers , 16, 18.earchers  indicate T. taeniaeformis, there is a need of risk assessment of the rat as well as cat population in these two parishes. This study has found strong evidence to educate the community regarding proper maintenance of hygienic conditions in and around their dwellings to prevent the survival and proliferation of the rat population.The Grenadian community likes cats as a pet. However, these pets are not always confined inside the home resulting in roaming behavior near and around the residential areas. The population of cats in the study areas of St. George's and St. David is not known. Since rats are the intermediate host and are the final host for


Haemonchus contortus, have caused serious losses in livestock production, particularly after the emergence of resistance to conventional anthelmintics. The search for new anthelmintic agents, especially those of botanical origin, has grown substantially due to the perspective of less contamination of meat and milk, as well as other advantages related to their cost and accessibility in less developed countries. The aim of this study was to evaluate the in vitro anthelmintic activity of essential oils of the plant species Citrus aurantifolia, Anthemis nobile and Lavandula officinalis against the main developmental stages of the parasite H. contortus.Infections of sheep with gastrointestinal parasites, especially C. aurantifolia, A. nobile and L. officinalis, respectively. Different concentrations of each oil were tested in vitro for their capacity to inhibit egg hatching (EHT), larval development (LDT) and adult worm motility (AWMT) using a multidrug-resistant strain of H. contortus (Embrapa 2010). The IC50 values obtained for the oils of C. aurantifolia, A. nobile and L. officinalis were 0.694, 0.842 and 0.316 mg/ml in the EHT and 0.044, 0.117 and 0.280 mg/ml in the LDT, respectively. The three oils were able to inhibit adult worm motility completely within the first 8–12 h of observation in the AWMT.Plant species were selected based on substantial ethnopharmacological information. Analysis of the composition of each oil by gas chromatography coupled to mass spectrometry (GC-MS) demonstrated the presence of limonene (56.37%), isobutyl angelate (29.26%) and linalool acetate (35.97%) as the major constituents in H. contortus. Furthermore, this study is of ethnopharmacological importance by validating the anthelmintic activity of the oils studied. Although new experiments are necessary, these data contribute to the development of pharmaceutical-veterinary products for sheep farming by opening up new therapeutic possibilities against gastrointestinal infections caused by H. contortus.The present results demonstrate significant anthelmintic activity of the three oils against the different developmental stages of  Haemonchus contortus , completely inhibiting motility within the first 8–12 h of observation. In addition, a marked overall inhibition of motility of 50% or higher was observed for all oils within the first 2–4 h of the experiment , is advantageous for the choice of substances to be tested in vivo . These aontortus .Haemonchus contortus strain, Embrapa 2010, is known to be resistant to the main classes of commercially available anthelmintic drugs, such as benzimidazoles, macrocyclic lactones and imidazothiazoles [For the development of new anthelmintic agents, it is important that the parasite species, and even its strain, be the most representative of the livestock in reality, especially when this type of economical activity faces different challenges . Haemonchiazoles . In facthiazoles .Essential oils consist of a mixture of volatile and lipophilic compounds with a strong and characteristic odor. Terpenes and terpenoids are their main chemical constituents. These compounds differ by the presence of an oxygen atom in terpenoids and a methyl radical in terpenes . In viewH. contortus, have been described for a large number of essential oils such as those of Croton zehntneri [Eucalyptus staigeriana [Cymbopogon martinii and Mentha piperita [Lippia sidoides [Thymus vulgaris [Human knowledge of essential oils originated in the pre-Christian era . Anthelmehntneri , Eucalypigeriana , Cymbopopiperita , Lippia sidoides  and Thymvulgaris .L. officinalis was more effective in the EHT than the oils of C. aurantifolia and especially of A. nobile. In fact, the IC50 value of the L. officinalis oil was about 2.0–2.5 times lower than that of the other two oils tested . Limonene is characteristic of species of the genus Citrus [L. officinalis (0.52%)  Table . Myrceneis Table . Borneolis Table . Except Lavandula [L. officinalis   and isoamyl angelate (15.27%) (Table Linalool acetate and trans-sabinene hydrate are the major constituents of plants of the family Lamiaceae, especially in the genus avandula . These wH. contortus. For example, limonene is found in the oils of different plants with anthelmintic activity, such as Lippia sidoides, Cymbopogon martinii, Mentha piperita and Eucalyptus staigeriana [Citrus sinensis, causing 100% inhibition of H. contortus eggs and larvae in vitro at most concentrations tested [Ascaris lumbricoides [A. nobilis and L. officinalis. This compound is well known to be effective against the egg and larval stage of H. contortus [In addition to the representative importance of major constituents of each oil studied, some of these compounds possess known anthelmintic properties, highlighting their potential importance in final anthelmintic activity. The evidence for the activity of such compounds can be direct or indirect, as observed for igeriana , 29, 34.s tested . Linalooontortus .H. contortus because of the advantages of these agents, such as low cost and easy access, especially in less developed countries with biodiversity [In view of the economic impacts of gastrointestinal infections on sheep farming, which are increasingly aggravated by anthelmintic resistance, the identification and development of new therapeutic options for prevention, control or eradication of this problem is essential , 36. In iversity . Since biversity . Finallyiversity .C. aurantifolia, A. nobile and L. officinalis as anthelmintics. This is the first study testing the essential oils of these plants against H. contortus, highlighting their ethnopharmacological use and pharmaceutical-veterinary value.The application of ethnopharmacological/chemotaxonomic-based strategies is important to optimize studies of botanical anthelmintic agents in order to increase the success of developing new plant-derived or herbal agents of interest for the pharmaceutical-veterinary industry . AlthougC. aurantifolia, A. nobile and L. officinais essential oils against the main developmental stages of the parasite H. contortus. It highlights the importance of promising medicinal plants and theirs chemical compounds for the development of new potential anthelmintic agents.This study reveals the in vitro effects of


Mathematical modeling has become a valuable tool for the analysis of dynamics of infectious disease and for the support of control strategies development in recent years. This work highlights the conceptual ideas and mathematical tools needed for infectious diseases modeling. The main convergence of this was on the dynamics of infectious diseases, the analysis of transmission patterns in various populations, and methods to assess the effectiveness of control strategies such as HIV, childhood infections, influenza, and vector borne infections. It was concerned with qualitative behaviors of infectious disease model. The qualitative behavior of model includes positivity, uniqueness, local stability, global stability, bifurcation analysis, control of diseases, and existence of solutions. This study provided a platform for the discussion of the major research challenges and achievements on qualitative behaviors of infectious diseases and their control. Due to the availability of a lot of applications of this study, many authors contributed.G. R. Phaijoo and D. B. Gurung demonstrated that dengue is spreading in new areas due to people movement. They considered a multipatch model to assess the impact of temperature and human movement on the transmission dynamics of dengue disease. Dynamics of vector and host populations are investigated with different human movement rates and different temperature levels.N. Pipatsart et al. discussed adaptive random network models to describe human behavioral change during epidemics and performed stochastic simulations of SIR epidemic models on adaptive random networks.C. Burgess et al. derived risk-based immunization by deployment to polio-endemic regions, which is sufficient to prevent transmission among both deployed and nondeployed US military populations.T. Tilahun et al. analyzed a compartmental nonlinear deterministic mathematical model for the typhoid fever outbreak and optimal control strategies in a community with varying population.L. Worden et al. extended an approximation technique of the long-term behavior of a supercritical stochastic epidemic model, using the WKB approximation and a Hamiltonian phase space, to the subcritical case.S. Anis et al. discussed some of the genetic properties on the basis of algebra.Finally, A. Miao et al. proposed a stochastic SIR model with vertical transmission and vaccination. They showed that large noise can lead to the extinction of infectious diseases, which is conducive to epidemic diseases control.Gul ZamanIl H. JungDelfim F. M. TorresAnwar Zeb


We would like to issue a corrigendum concerning the following points. Please note that these corrections are not related to the analysis of the data and the presentation of the results, nor they affect in any way the conclusions of our article. We deeply apologize for any inconvenience caused.1. The following authors with the affiliations shown below should be added to the author list of the paper, between C. Sanz (first author) and E. Tagliazucchi (last author) in the following order:federicozamberlan@hotmail.comFederico Zamberlan. Affiliation: Departamento de Física, Universidad de Buenos Aires, Argentina. Email: research@erowid.orgEarth Erowid. Affiliation: Erowid Center, Grass Valley, CA, USA. Email: research@erowid.orgFire Erowid. Affiliation: Erowid Center, Grass Valley, CA, USA. Email: Thus, the author list of the article should read as follows:“C. Sanz, F. Zamberlan, E. Erowid, F. Erowid, E. Tagliazucchi”The reason for this correction is the re-evaluation of the merits of the contributors that were originally mentioned in the “Acknowledgments” section of the manuscript. After friendly discussions with E. Erowid and F. Erowid, we reached the agreement that their role in the creation, curation and interpretation of the online database, as well as in the ensuing scientific discussion, should be acknowleged with co-authorship. The addition of F. Zamberlan to the author list is based on similar contributions, related to our local version of the database (creation and curation) and on his scientific input, in all cases fundamental for the research conducted.2. The following sentence should be added in the Materials and Methods” section, “Corpora Selection” sub-section, right before the sentence: “We discarded reports that resulted…”:https://erowid.org/general/about/about_copyrights.shtml).”“Our research relied upon Erowid's reviewed and edited collection of experience reports, and followed Erowid's terms of use that require researchers to coordinate with Erowid's research team in order to avoid misinterpretations of their data (The reason for this correction is to encourage future researchers to comply with Erowid's terms of use. Such compliance is necessary to acknowledge the efforts of the Erowid team, and to avoid misinterpretations of the reports, thus improving the overall scientific accuracy of the resulting manuscripts.3. The “Author contribution statement” section should be changed as follows to reflect the changes in authorship:“CS and ET analyzed the data. ET designed the study and wrote the paper. EE and FE designed the Erowid experience report collection system and have managed the collection of psychoactive-related experience reports since 1995. FZ contributed to data analysis, data interpretation, and to the creation and curation of a local database of experience reports.”4. The “Acknowledgments” section should be changed as follows to reflect the changes in authorship:www.erowid.org and www.dreamjournal.net.”“ET was supported by a Marie Skłodowska-Curie individual fellowship. FZ was supported by a CONICET doctoral fellowship. We acknowledge insightful discussions with Facundo Carrillo, Mariano Sigman, and Diego Fernandez Slezak. We also thank the curators and contributors of We apologize for these mistakes, which do not change the scientific conclusions of the article in any way. All authors approved this corrigendum.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Oral metastases occur more commonly in bone, but can also manifest in soft tissues and eventually resemble a reactive lesion. Few cases of oral metastases mimicking reactive lesions in soft tissues have been reported to date.We report a metastasis of gastric carcinoma (GC) to the oral mucosa without bone involvement in a 43 yom clinically and microscopically mimicking a reactive lesion. The patient related that the lesion had 1 month of evolution, and the ulcerated area suggested the lesion was related to trauma.The histopathological examination of the lesion revealed an exuberant granulation tissue with few neoplastic cells, and the initial diagnosis of pyogenic granuloma was considered. In a second analysis, clusters of clear cells morphologically similar to degenerating mucous cells or macrophages, positive for Cytokeratin (CK)-20, and CDX2 were found. At the moment, it was confirmed the presence of a primary GC in the patient.A palliative radiotherapy/chemotherapy was started.However, the patient died 3 months after the diagnosis of oral metastasis.This report highlights the importance of careful clinical and microscopic examinations in cases of oral metastasis that may mimic a reactive lesion. It is also important to consider that approximately 25% of the oral metastases comprise the first evidence of an undiscovered malignancy at a distant site.,4Metastatic tumors of the mouth represent only 1% of all malignancies affecting this region. Usually oral metastases involve the jawbones and more rarely the soft tissues. Metastases in oral soft tissues usually manifest as ulcerated lesions or masses causing swellings. In the mouth, a few cases of metastases resembling pyogenic granuloma were reported, and it seems that this type of presentation is more common in the skin.,6Concerning the oral mucosa, the most common sites for metastasis are the gingiva, followed by the tongue and with less frequency the remaining oral soft tissues.,2 Oral metastases from gastric adenocarcinoma (GC) are rare, although this malignancy represents the fourth most common cancer in man and the second most frequent cause of human cancer death.,8The major primary sites presenting metastases to the mouth include lungs, kidney, liver, and prostate for men, and breast, uterus, ovaries, kidney, and colorectum for women.In this report, we describe a metastatic oral mucosa lesion from gastric adenocarcinoma, clinically and microscopically resembling a pyogenic granuloma.2The authors read the Helsinki Declaration and followed its guidelines in this study.Our service received a biopsy of a 43-year-old male for evaluation of an exophytic ulcerated mass involving the posterior region of the right mandible, with clinical hypothesis of a pyogenic granuloma or peripheral giant cell lesion. According to the patient, the lesion had 1 month of evolution, and the ulcerated area suggested the lesion was related to trauma -7, CK20, and CDX2, many positive for Ki-67, and negative to CD68 Fig. . After a3; however, it uncommonly causes distant metastases, usually involving bones of the vertebrae, pelvis ribs, and skull,,10 and rarely the gnathic bones. In this report, we describe a metastasis of a gastric adenocarcinoma into the buccal mucosa of a 43 yom that initially was considered as pyogenic granuloma. The lesion was ulcerated and exophytic involving the molar area of the right mandible. Histopathological examination of the biopsy disclosed an exuberant granulation tissue covered by fibrinopurulent membrane, with few clear cells considered as macrophages. A second analysis was performed, and small clusters of clear cells presenting a degenerative aspect were found. By immunohistochemistry, it was concluded that these clear cells represented metastasis of a GC. Isolated clear cells of the first analysis also proved to be neoplastic gastric cells. The diagnosis was clinically confirmed as the patient presented a primary adenocarcinoma of the stomach. This case illustrates that oral metastases may mimic reactive lesions both clinically and microscopically urging the necessity of careful examination. The first analysis displayed mainly granulation tissue, as the lesion was ulcerated due to dental trauma and this is common in many lesions of the mouth, indicating necessity of deeper biopsies. In fact, small and superficial biopsies are common for oral lesions, frequently difficulting the correct diagnosis.GC is the fourth most common cancer in humans and the second cause of cancer death,12 but glandular pattern and nests of epithelial gastric cells also have been reported.–15 However, on the present case predominated an exuberant granulation tissue, suggesting a reactive lesion as pyogenic granuloma, and only on second analysis clusters of clear cells were more evident that represented metastasis of a gastric adenocarcinoma.Metastatic GC to the oral cavity tends to present signet ring cell, These lesions showed no bone involvement, as the present case, indicating a probable predilection of GC to metastasize to soft tissues of the oral cavity. Sauerborn et al have revised the cases of gastric metastatic carcinoma to oral soft tissues. In 18 cases reported, 10 occurred in the mandibular region, whereas 6 involved the maxilla, one each the tongue and soft palate. However, it was not reported that these lesions resembled benign reactive lesions. Furthermore, the histologic aspect of these reported cases was all compatible with adenocarcinomas, whereas in the present case only few cells were initially found immersed in a granulation tissue. To our knowledge, this is the first report of a metastatic GC to oral cavity mimicking a pyogenic granuloma.Metastatic GC to the oral cavity was previously reported including gingiva, soft palate, and soft tissues of the mandible. In the oral soft tissues, angiogenesis associated with chronic inflammation is very common, and this can facilitate the metastatic process, particularly the gingiva and adjacent tissues. Other studies correlated tooth extractions as a promoting factor in the metastatic process, supported by the same hypothesis of chronic inflammation caused by this procedure.The mechanisms of the metastatic process to the oral soft tissues are not fully understood. The ability of the primary tumor to stimulate angiogenesis and invade adjacent vessels has been considered crucial for spreading and distant colonization by the neoplastic cells, resulting in metastasis. However, this process comprises a complex biological cascade as the neoplastic cells acquire high ability for motility and invasion, survival, and proliferation.Overall, a bad prognosis is expected for metastatic neoplasms to the oral cavity, and death is common after a few months of the initial diagnosis. However, palliative treatment is important to improve life quality and pain relief.In conclusion, oral metastasis of gastric adenocarcinoma is rare, and it may eventually mimic a reactive process, difficulting the diagnosis as illustrated in this report.Conceptualization: C.D. Soares, L.M.R. Paranaiba, M.G.F. Carvalho.Data curation: C.D. Soares, M.G.F. Carvalho.Formal analysis: M.G.F. Carvalho.Funding acquisition: J. Jorge, O.P. Almeida.Investigation: C.D. Soares, M.G.F. Carvalho.Methodology: C.D. Soares, L.M.R. Paranaiba, M.G.F. Carvalho.Project administration: J. Jorge, M.G.F. Carvalho, O.P. Almeida.Resources: M.R. Melo-Filho.Supervision: B.A. Rocha.Writing – original draft: C.D. Soares, M.G.F. Carvalho.Writing – review & editing: J. Jorge, M.G.F. Carvalho, O.P. Almeida.


The institutional affiliation number 4 for the sixth author, Tero A. H. Järvinen, is incorrect. The correct affiliation is: Departments of Orthopedics & Traumatology and Anatomy, Medical School, University of Tampere and Tampere University Hospital, Finland.In addition, funding organizations for the sixth author were incorrectly omitted from the Funding Statement. The following sentence should be included with the Funding Statement: "T.A.H.J. is funded by the Academy of Finland, Instrumentarium Research Foundation, Sigrid Juselius Foundation and Finnish Medical Foundation."


Mycobacterium abscessus, a rapid-growing non-tuberculous mycobacterium, has been the cause of sporadic and outbreak infections world-wide. The subspecies in M. abscessus complex  are associated with different biologic and pathogenic characteristics and are known to be among the most frequently isolated opportunistic pathogens from clinical material. To date, the evolutionary forces that could have contributed to these biological and clinical differences are still unclear. We compared genome data from 243 M. abscessus strains downloaded from the NCBI ftp Refseq database to understand how the microevolutionary processes of homologous recombination and positive selection influenced the diversification of the M. abscessus complex at the subspecies level. The three subspecies are clearly separated in the Minimum Spanning Tree. Their MUMi-based genomic distances support the separation of M. massiliense and M. bolletii into two subspecies. Maximum Likelihood analysis through dN/dS  identified distinct genes in each subspecies that could have been affected by positive selection during evolution. The results of genome-wide alignment based on concatenated locally-collinear blocks suggest that (a) recombination has affected the M. abscessus complex more than mutation and positive selection; (b) recombination occurred more frequently in M. massiliense than in the other two subspecies; and (c) the recombined segments in the three subspecies have come from different intra-species and inter-species origins. The results lead to the identification of possible gene sets that could have been responsible for the subspecies-specific features and suggest independent evolution among the three subspecies, with recombination playing a more significant role than positive selection in the diversification among members in this complex. Mycobacterium abscessus is a rapidly growing species of non-tuberculous mycobacterium (NTM). In 2011, this species was divided into two subspecies, M. abscessus subsp. abscessus and M. abscessus subsp. bolletii . We included strains from different parts of the world to increase the likelihood of identifying subspecies-specific characteristics instead of just clonal traits.Of the 243 genomes examined, 12 were from clinical strains isolated between 2009 and 2011 in the Clinical Microbiology Laboratory of University of Malaya Medical Centre (UMMC), Kuala Lumpur, Malaysia. These strains were subjected to whole genome shotgun sequencing using the Illumina Genome Analyzer IIX platform, as described previously . For thein silico inter-subspecies and intra-subspecies comparisons, a single pseudo-contig for each of the 243 genomes was generated by using “NNNNNCACACACTTAATTAATTAAGTGTGTGNNNNN” as a linker between contigs. The insertion of this 36 bp pseudo-molecule introduces a stop codon in any of the six open reading frames, and hence will not affect the annotation of each genome , assuming Chi-square distribution with degree of freedom of 1  is greater than one. The genes with significant traces of positive selection were further analyzed with the Bayes Empirical Bayes (BEB) method, to retrieve the positions of amino acids on the genes that were substituted owing to selection forces  in CodeML of PAML on the dataset . In the dom of 1 . Genes wn forces . The genn forces .2 (Max χ2)  , and Pai(Max χ2) . NSS is M. abscessus and M. massiliense were selected as representatives for comparisons. The selection was based on the mean MUMi (indicating the average genomic distance between paired strains in a group), with five genomes each from the strains with the highest, median and lowest mean MUMi values. The intention of the selection was to maximize the inclusion of strains with high genetic diversities. Since there were only 16 M. bolletii strains, all M. bolletii genomes were included for the genome-wide alignments (Supplementary Table S1). (2) Only the LCBs with a minimum size of 500 bp which were shared by all the strains were used for the analysis. An in-house Biopython script was written to separate each LCB from the XMFA alignment file. (3) The sequence of each LCB was then identified from the genomes that were not used in the Mauve alignment. Each of the resulted LCB was manually curated to remove artifacts, and concatenated to form super-sequences. The super-sequences and a PhyML generated super-sequences SNPs tree were subjected to recombination study in the ClonalFrameML  as defined by the progressiveMauve algorithm . Owing tlFrameML .M. abscessus complex isolated from multiple countries. The subspecies for each of the 243 strains was identified in silico using the concatenation of the 50 proposed informative genes previously reported . The MST illustrates the relationships of data based on the shortest possible distance between two data points. Strains from the same subspecies would have low dissimilarities and short genetic distances from each other, and hence, are expected to form a cluster. Based on this classification, 16 strains in our study clustered as M. bolletii, 138 as M. massiliense, and 89 as M. abscessus.The genomes used in this study comprised the three known subspecies of reported . The resM. massiliense strains compared to M. abscessus and M. bolletii. These observations suggest that the evolution of the M. abscessus complex is not clonal but is driven by the genetic characteristics of each subspecies within the complex.On the MST, the size of each circle corresponds to the number of genomes within respective nodes. Generally, strains within the same subspecies were clustered together without passing through any node from other subspecies. The MST was constructed with 100 bootstrap replications and each bootstrapping tree was consistently observed to have 110 nodes. All edges were supported with high bootstrap replication values at a minimum of 90% (represented in solid lines), except for 11 edges with lower bootstrap values of 50–70% (represented in dotted lines). The lower confidence edges were distant from the edges connecting the different subspecies. Hence, there were no strains with questionable subspecies status. No sign of geographical clustering among the strains was observed. The MST also highlighted the greater diversity among Supplementary Table S2). The inter-subspecies interquartile range for M. bolletii – M. massiliense, M. bolletii – M. abscessus, and M. abscessus – M. massiliense were 0.2326, 0.2072, and 0.2222, respectively. Hence, the genomic distances between M. massiliense and M. bolletii were longer than the distances between M. massiliense and M. abscessus or M. bolletii and M. abscessus. The taxonomic separation of M. massiliense from M. bolletii has been proposed by several research groups based on the analysis of core sequences with ANI as well as linear and network phylogenies , the MUMi of M. bolletii skewed toward the higher values of the box, reflecting a relatively conserved genetic content in this subspecies. In contrast, the MUMi of M. abscessus was skewed toward the lower values in the boxplot, indicating that the majority of the M. abscessus strains have longer genomic distances from each other. The MUMi of M. massiliense, however, showed a symmetrical distribution, indicating the presence of roughly equal numbers of conserved and diverse strains.The intra-subspecies interquartile range of MUMi was 0.0998 in M. massiliense, 14 in M. abscessus and 18 in M. bolletii, that could have undergone positive selection , during the course of divergence among the three subspecies.Our Maximum Likelihood analysis through dN/dS calculation on each of the 1,090 non-paralogous core genes in our dataset supports the presence of genes that have undergone selective pressure. It has been said that the dN/dS ratio is not sensitive enough to detect gene sets that have undergone positive selection among closely related species . However2) and a p-value cutoff at 0.01, only four out of 18 positively selected genes in M. bolletii (Supplementary Table S3) and 2 out of 14 genes in M. abscessus (Supplementary Table S4) were identified as having undergone recombinations. None of the positively selected genes in M. massiliense (Supplementary Table S5) showed evidence of recombination.Based on the results of the three independent recombination methods  in positive selection analyses . On the Acinetobacter baumannii , and probable endonuclease activity .In each M. abscessus complex, suggesting the occurrence of recombination events  were detected from the 1,407 LCBs generated by progressiveMauve. The lengths of recombinations ranged from 2 to 34,082 bp. The ratio of rates at which polymorphisms were introduced by recombination and mutation was R/theta = 0.14, whereas the ratio of recombination and mutation (r/m) was 1.99. Hence, recombination caused two times more polymorphism than mutation. From the illustration in Figure 3, extensive recombination was observed in the three putative ancestral nodes leading to the three subspecies. Since there are long evolutionary distances separating the three putative ancestral nodes, it is not known whether the accumulation of the recombinations occurred after the subspeciation event or was responsible for the independent emergence of the three subspecies.Apart from Darwinian evolution, recombination has been proposed as the alternative event for evolution . Recent n events . In thisM. massiliense (r/m = 2.17), with almost equal effect of substitutions and mutations in M. abscessus (r/m = 1.41) and M. bolletii (r/m = 0.95). We predicted 675 recombination events in M. abscessus, 387 in M. bolletii and 786 in M. massiliense. The analysis on recombination segments identified the origins of approximately 23% of total recombination segments in M. abscessus , 16% in M. bolletii  and 17% in M. massiliense . There were 33 segments in M. abscessus, 4 in M. bolletii and 11 in M. massiliense that could not be confirmed to be from an intra- or inter- subspecies origin. The segments that could not be found within the M. abscessus complex might have been acquired from foreign origins. However, we did not find insertion sequences (ISs) that could have contributed to the recombination events found in the M. abscessus complex. Neither did we find any CRISPRs that could have prevented the entry of foreign genetic material at the recombination sites. It is possible that novel genetic mechanisms are involved in these recombination events.Besides computing the whole population genome-wide recombination structures, subspecies- specific recombinations were also investigated by r/m estimations. The results suggest a higher recombination impact among Salmonella variants (Mycobacterium tuberculosis strains (M. abscessus complex which differ in their drug susceptibility and invasiveness, with M. massiliense reported to be the least associated with antibiotic resistance but the most frequently isolated from clinical material, and M. bolletii being the most frequently associated with multiple antibiotic resistance but the least frequently isolated from clinical specimens (Speciation genomics has contributed immensely to the understanding of bacterial evolution and diversity . It has variants  and drug strains . In thispecimens . We trieM. massiliense cluster, there is a wider spread of genotypes than in the other two subspecies. This feature indicates higher allelic variability among M. massiliense genomes. In contrast, M. bolletii showed the least allelic variation. Supported by our data on MUMi evaluation of both core and accessory genomes, the low median intra-subspecies index indicated that M. bolletii is conserved compared to M. massiliense and M. abscessus. This could be reflected in its low prevalence in infections. Using the same data, the inter-species distance appears longer between M massiliense and M. bolletii than between either of them and M. abscessus. This provides another reason to consider M. massiliense and M. bolletii as distinct subspecies.The MST showed the strains to be clustered according to the subspecies they belong to rather than by their countries of origin. Within the M. abscessus complex has been discussed by other researchers (The impact of the accessory genome on the evolution of the earchers . In thisM. massiliense (M. abscessus complex.From the ratio of synonymous and non-synonymous mutations and the evaluation of amino-acid changes based on BEB, we speculated site-specific positive selection resulting in amino acid substitutions. The biological significance of these substitutions is not known, but two proteins involved, the 30S ribosomal protein S3 and thymidylate kinase, were previously found to contain specific signatures for the classification of siliense . We woulM. abscessus complex is driven more by recombination than positive selections. Interestingly, our data revealed three independent recombination landscapes in the three subspecies. In a previous study (M. abscessus was the highest among the three subspecies. In the current study, M. massiliense appeared to be the most highly affected by recombination, followed by M. abscessus and M. bolletii, in descending order. The difference could be due to the use of different datasets. M. massiliense compared to that in M. abscessus is also supported by our previous network phylogenomics analysis (M. abscessus complex evolution for pathoadaptation. M. massiliense strains isolated from different countries (M. massiliense strains could also be the result of horizontal transfer and recombination involving common genetic determinants for infectivity. This could illustrate the advantage conferred by a high recombination rate on the pathoadaptation of M. massiliense.Our overall results suggest that the evolution of the us study , the impanalysis . Neverthountries  and thisountries . On the M. abscessus complex has been shaped by recombination more than Darwinian selection, and that each subspecies of the complex appears to have evolved independently. The proteins we identified to have undergone positive selection and recombination are expected to facilitate future studies on the functional significance of proteins in the M. abscessus complex.Our findings suggest that the diversity of the Planned and conceived the experiments: JLT and YFN. Performed the experiments: JLT and YFN. Analyzed the experiments: JLT, KPN, CSO, and YFN. Drafted the manuscript and contributed to the data interpretation: JLT, KPN, CSO, and YFN. All authors read, critically revised and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.