AbstractSternolophus Solier, 1834 was examined in this study using 60 morphological adult characters, eight of them continuous and 52 discrete. The cladistic analysis resulted in a single most parsimonious tree with two major subclades corresponding, respectively, to species previously assigned to the subgenera Sternolophus s. str. Solier and Neosternolophus Zaitzev, although they are not re-instated. The species groups S.angolensis  and S.solieri Castelnau, 1840 are recovered as monophyletic. The biogeography and diversification of the species of Sternolophus are briefly discussed.The phylogeny of the hydrophilid genus  Sternolophus Solier, 1834 is widely distributed in the tropics of the Old World, with only few species occurring in the temperate zones. In a recent taxonomic revision of the genus by The genus PageBreakSternolophus has been poorly studied. Sternolophus s. str. Solier, 1834 and Neosternolophus Zaitzev, 1909. His classification was based on the absence or presence of an emargination on the anterior clypeal margin. Although this subdivision was accepted by Neosternolophus to generic rank based on the emargination of the anterior clypeal margin, but this change was later opposed by Sternolophus species were also studied by Sternolophus in his analysis of the subtribe Hydrophilina which resulted in the monophyly of the subgenusSternolophus s. str. and the lack of resolution for species of Neosternolophus.The phylogeny of S.angolensis  and S.solieri Castelnau, 1840) based on highest morphological similarity and without including a phylogenetic approach. These authors considered S.angolensis, S.inconspicuus , S.mundus  and S.solitarius Nasserzadeh and Komarek, 2017 as members of the angolensis group, and placed S.angustatus , S.elongatus Schaufuss, 1883, S.mandelai Nasserzadeh and Komarek, 2017, S.rufipes , and S.solieri in the solieri group. They left the remaining species , and S.prominolobus Nasserzadeh and Komarek, 2017) ungrouped.Sternolophus is provided, based on a cladistics analysis of adult morphological characters. Considering the phylogenetic results, the biogeography and diversification of the species are briefly discussed.Here the first comprehensive phylogenetic analysis of the genus Taxon sampling. More than 4000 specimens in all the 17 species of Sternolophus were studied as ingroup, and Hydrocharaflavipes, belonging to the tribe Hydrophilini, was included as outgroup. A total of 271 specimens were measured. The specimens were obtained on loan from the following institutions and collections:AEZS coll. A. Short, University of Kansas, Lawrence, KS, USACBSU Collection of Department of Biology, Shiraz University, IranHMIM Hayek Mirzayans Insect Museum, Tehran, IranPageBreakISNB Institut Royal des Sciences Naturelles de Belgique, Bruxelles, BelgiqueMNHN Muséum National d’Histoire Naturelle, Paris, FranceMNHUB Museum der Alexander Humboldt Universität, Berlin, GermanyNHML Natural History Museum, London, UKNMB Naturhistorisches Museum Basel, Basel, SwitzerlandNMW Naturhistorisches Museum Wien, Vienna, AustriaNRM Swedish Museum of Natural History, Stockholm, SwedenOUMNH Oxford University Museum of Natural History, UKSAMA South Australian Museum, Adelaide, AustraliaSMTD Staatliches Museum für Tierkunde, Dresden, GermanyZMUC Zoological Museum University of Copenhagen, DenmarkThe examined specimens are listed in Appendix Preparation for morphological studies. To study the male genitalia, the aedeagus was extracted and macerated in lactic acid for at least four days to become hydrated and cleared before examination. Bursa copulatrix, spermatheca, and spermathecal gland were also dissected    with ‘trThe synapomorphic characters and character states are mapped on the single most parsimonious cladogram . Branch support was calculated by bootstrap , jack-knThe consistency and retention indices  of discrPageBreakPageBreakPageBreakof discrete characters only  resulted in 36 most parsimonious trees of 110 steps. The consensus trees using both strict and majority-rule methods were different from previous trees in the position of the species in clade B  resulted in a single most parsimonious tree of 146.130 steps Fig. . When sue B Fig. . AnalysiSternolophus species are divided into two major monophyletic clades, B and G, with 6 and 11 species respectively. Clade B contains S.decens as sister to clade C that is composed of five species, S.solieri, S.rufipes, S.angustatus, S.mandelai, and S.elongatus. Clade B is supported by five characters , although it is weakly supported statistically. Except for the elongated spine on the metaventrite (37: 1), the characters sustaining this clade were homoplastic. The topology of clade B was slightly different in analysis C .As shown in the single most parsimonious tree obtained with analysis A Fig. , the exas C Fig. , and thes C Fig. , whereass C Fig.  S.decenPageBreakS.australis is sister to clade I, whereas S.immarginatus is sister to the clade formed by S.australis and clade I. In all analyses, clades K, L, M, and N were found to be monophyletic with the same configuration. These clades are supported by one, two, three, and three synapomorphies, respectively  (Nasserzadeh & Komarek 2017). In all analyses, the topology of clade G remained consistent except for slight changes in clade M and variable support for clades G and I  Unreliable topology of clade B in different analyses and absence of support for its monophyly as well as monophyly of the subclades. ii) Questionable position of S.decens within clade B. Sternolophusdecens was included in the subgenusSternolophus s. str. by S.rufipes and S.solieri by PageBreakHowever, it was recovered in a monophyletic clade together with S.marginicollis (and some unidentified Sternolophus species) by subgenusNeosternolophus by S.decens was recovered as sister to clade C. The species of this clade   Fig.  were groHydrophilina based on adult-morphological characters, found evidence for monophyly of the subgenusSternolophus s. str., but the species formerly grouped in the subgenusNeosternolophus were unresolved and formed a basal polytomy within the genus. In our analysis, on the contrary, strong evidence was found for monophyly of Neosternolophus, whereas monophyly of Sternolophus s. str. is more questionable for the reasons mentioned above.S.solitarius, S.mundus, S.inconspicuus and S.angolensis) grouped by angolensis species group based on morphological similarities, are resolved here as clade M confirming their close relationship, although weakly supported (Fig. Finally, the four species (ted Fig. .Biogeography and diversification. In Figure solieri species group distributed in the Afrotropical, Palaearctic and Oriental regions. Distribution of S.decens overlaps with those of clade D. On the other hand, most members of clade G have an Oriental-Australasian distribution. The exceptions are representatives of the angolensis species group, with S.solitarius, S.mundus, and S.angolensis restricted to the Afrotropical Region whereas S.inconspicuus is widely distributed in the Oriental Region to the eastern boarder of the Palaearctic Region. Sternolophusinsulanus and S.jaechi are two sister species with insular distribution in the Malay Archipelago (see Appendix Sternolophus, dispersing toward Australia in the Oligocene/Miocene. There are many New Cenozoic fossil findings of taxa closely related to Sternolophus in Europe and North America (e.g. S.rufipes from the Early Pliocene of the Tsubusagawa Formation in Japan (Sternolophus in the Old World, i.e. without protruding into northern Asia, Europe, Tasmania and New Zealand (ica e.g. , 2010b, in Japan . The cur Zealand , which wPageBreak


Cronobacter species was incorrectly described as ten. Actually, Cronobacter pulveris, Cronobacter helveticus and Cronobacter turicensis were removed from the genus in 2014.In the first paragraph in the “Introduction” of this article, the number of Cronobacter spp. is currently considered to consist of 7 species: Cronobacter sakazakii, C. malonaticus, C. universalis, C. dublinensis, C. muytjensii, C. condiment, and C. zurichensis (Stephan et al., C. sakazaii is one of the two group 1 clinically relevant species that form the majority of the clinical isolates. The authors regret this error.The original sentence should be corrected as follows: 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.


Helicobacter pylori is a causative pathogen of chronic gastritis, gastric ulcer disease, and gastric cancer. Humans are known to be a natural host for H. pylori and tend to acquire the pathogen before the age of 5 years. The infection may then persist lifelong if eradication therapy is not applied. One of the modes of transmission of H. pylori is between family members, and therefore, the presence of infected family members is an important risk factor in children. However, other environmental factors have not been fully analyzed. The present study was performed to clarify whether and to what extent intestinal microbiota affect H. pylori intrafamilial infection. The fecal specimens from H. pylori-infected infants and H. pylori-infected and non-infected family members were collected in cohort studies conducted by Sasayama City, Hyogo Prefecture from 2010 to 2013. In total, 18 fecal DNA from 5 families were analyzed. Samples were amplified using 16S rRNA universal primers, and the amplicons were sequenced using the Ion PGM system. Principal-coordinate analysis demonstrated that there was no difference in intestinal microbiota between H. pylori-positive and H. pylori-negative groups. In intrafamilial comparison tests, the Manhattan distance of intestinal microbiota between the H. pylori-infected infant proband and H. pylori-negative mother was nearest in the family with low intestinal microbial diversity. However, in the family with the highest intestinal microbial diversity, the nearest Manhattan distance was shown between the H. pylori-infected infant proband and H. pylori-infected mother. The results in this study showed that the composition of the intestinal microbiota was very similar between members of the same family, and as such, colonization with organisms highly similar to the infected parent(s) may be a risk factor for H. pylori infection in children. Helicobacter pylori and more than half of the world’s population are infected with this microorganism. Long-term infection with H. pylori significantly increases the risk of developing site-specific diseases, such as peptic ulcer disease  and real-time polymerase chain reaction detection of H. pylori DNA by the 16S rRNA gene of H. pylori-targeted primers were used  according to the manufacturer’s instructions with some modification . Two hunEach DNA specimen was amplified using the Ion 16S™ Metagenomics Kit . The amplicons were purified and prepared for the sequencing library by using the Ion Plus Fragment Library Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. The library was sequenced by using the Ion PGM system (Thermo Fisher Scientific) and the Ion PGM Hi-Q sequencing kit following the protocol of the kit , 18.The operational taxonomic unit of each sequence was determined by the Ion torrent server with the Greengenes database . The min2 test was performed. A probability value  less than 0.05 was considered statistically significant.Statistical analysis was performed using the software KaleidaGraph . Mean values were compared among subjects using the Kruskal–Wallis test between all groups. For comparing categorical data, the χH. pylori-infected children and their family members were tested for microbiota analysis. The details are shown in Table H. pylori-infected mothers and three H. pylori-infected fathers, four H. pylori-uninfected siblings, and one each uninfected father and mother, as well as five index children were tested. All five H. pylori-infected index children had a H. pylori-infected mother and/or father but no H. pylori-infected sibling.The 18 fecal specimens from 5 H. pylori infection and age, the subjects were divided into H. pylori infected and non-infected, and the subgroups were also divided into parent and child groups. In these four groups of subjects, the abundance of Bacteroidetes or Firmicutes and the ratios of Firmicutes/Bacteroidetes were compared . H. pylori was not detected in H. pylori-negative and -positive subjects at all from fecal microbiota.The average relative abundance of each bacterial family, genus, and species was compared among the four groups. At the family level, abundances of  Figures A–C. At tn Figure D. Higher Figures E,F. At tn Figure G. Highers Figure H, and Cln Figure I. TuriciH. pylori status and age groups. As shown in Figure H. pylori-negative parents than in those from H. pylori-positive parents. These results remained significant even when the comparisons were performed at family and genus levels  of Manhattan distances highlighting similarities between fecal specimens of the 16 family members is shown in Figure PC1 and PC2 represent the first two highest discriminating axes. At both levels of species and family, family 2 was divided and separated from other families Figure . PCoA plHelicobacter positive index children in the five families. However, one (family 1) of four mothers was H. pylori negative, and the other four members were H. pylori positive.Manhattan distances at the bacterial family level between family members in each family are shown in Table H. pylori-positive parent to children in family 1, the distance between the H. pylori-positive parent to the index child  of human and animal subjects and that the eradication of H. pylori infection induces an increase in BMI  were shown between index and H. pylori-infected mothers (three cases) and father (only one case). These data showed that the similarity of microbiota was related to transmission of H. pylori infection.In this study, we also focused on the similarity of the index child and other family members, especially H. pylori-negative siblings and H. pylori-positive parents were compared to those between index children and H. pylori-positive parents and shown to be shorter with the exception of family 2, where all three family members were similar to each other. In family 5, the H. pylori-positive index child was similar to the H. pylori-positive father, and the H. pylori-negative sibling showed a similar microbiome to that of the infected mother implying father-to-child transmission.The Manhattan distances between H. pylori and that similarity of microbiota could be considered a risk factor. However, it is possible that the similarity in microbiota is also an effect of H. pylori infection. This possibility is worthy of further exploration.In conclusion, our data support the idea that the intestinal microbiota may contribute to intrafamilial transmission of This study was carried out in accordance with the recommendations of ethical guidelines for clinical research in Japan. All experiments were performed in accordance with the principles of Declaration of Helsinki with written informed consents. The study protocol (H22-047-02) was approved by the ethics committees of Kyorin University.Conceived and designed the experiments: TO, SKikuchi, SKamiya, and MO. Wrote the paper: TO, CZ, YL, and SKamiya. Metagenome analysis: TO, EN, FH, and HY. Analyzed the data: TO and HY. Contributed reagents/materials/analysis tools: SKurata and TH.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.


Manchurochelys manchoukuoensis. Several juvenile features are uncovered, such as a small and circular carapace , wide vertebral scales, and lateral carapacial fontanelles. In contrast to the adult morphology, which has an oval carapace, closed lateral fontanelles, and longer vertebrals 2–4, the juvenile of M. manchoukuoensis is more comparable to that of Sinemys lens, except for earlier occurrence of the well-ossified carapace of the latter. Differs from Changmachelys bohlini, and Ordosemys liaoxiensis, in which the circular carapace is relatively independent of ontogenetic age, and the lateral fontanelles are only closed in adult stage of O. liaoxiensis. Therefore, the trajectory of ontogenetic change appears to be highly diversified in the sinemydids.A small juvenile turtle is described from the Early Cretaceous Jehol Biota, shedding light on the juvenile morphology and ontogeny of  Manchurochelys manchoukuoensis, Ordosemys liaoxiensis, Liaochelys jianchangensis, Xiaochelys ningchengensis) have been discovered from the Jehol Biota and a large number of well-preserved fossils have been described , however, the neurals are varied in shape and size . The lasFollowing the neural row, two suprapygals are developed. The first suprapygal is distinctly smaller than the second one. However, it is proportionately smaller than the suprapygal I of PMOL-AR00008 . The pygAs a juvenile specimen, the costal plates are not fully ossified at their distal ends. Therefore, the lateral fontanelles are present between the costals and peripherals. In contrast, the fontanelles are closed in the adults , in which the vertebrals 2–4 are longer than wide . The firThe articulated cervicals form a S-shaped cervical series – 3. The The third cervical is relatively shorter than the axis. The neural spine of the third cervical is also broken off. Based on its remains, the neural spine is relatively reduced, and only present on the anterior half of the neural arch. As in the axis, the shelf-like ridge is developed which is parallel to the posterior margin of the neural arch. In the fourth cervical, the left side is somewhat damaged and the left prezygapophysis and postzygapophysis are broken off. The neural spine is low. The posterior margin of the neural arch is V-shaped as a result of the deeply divergent postzygapophyses. The V-shaped posterior margin is highlighted in the fifth and sixth cervicals . Due to Seven caudal vertebrae are preserved in articulation extending from the carapace. The posterior half of the first caudal is exposed. Its anterior part is still overlapped by the carapace. The transverse process extends more laterally than posteriorly. The second caudal is slightly displaced from its original position, and exposed in posterior view. The posterior articular facet of the centrum is a shallow concavity, implying a presence of an opisthocoelous or amphicoelous centrum. The neural canal is relatively smaller than the centrum. The third caudal is wholly exposed in dorsal view, although the posterior part of the neural arch is broken off. The prezygapophyses are nearly paralleled to each other. The articular facet is strongly oblique. The transverse process is positioned more posteriorly than on the first caudal. The process extends more posteriorly than laterally, and well beyond the level of the vertebral body. The transverse processes reduce their size gradually in the succeeding caudals, and appear to be absent in the sixth or seventh caudals. The neural spine is well preserved in the sixth caudal. The spine is low, broad and rounded.Another segment of the caudal series includes 12 vertebrae, possibly representing the distal caudals. This segment is displaced from the anterior portion. The first vertebra of the segment is strongly reduced, and is roughly half the length of the sixth caudal of the anterior portion. The complete number of caudals is therefore uncertain now. These caudals are also more or less damaged. They are exposed in lateral view, and have a distinctly curved ventral margin.In the forelimb, humerus bears an expanded distal end. The ectepicondyle foramen is open. The ulna and radius are slender and rod-like. They are subequal with a length of 10.4 mm. The ulna is relatively stronger than the radius. The manus is poorly preserved on both sides.The hindlimb is well preserved on the right side. The tibia and fibula are about 13.4 mm long. The tarsals are positioned between the tibia and fibula and metatarsus. However, a further identification of the tarsal elements is difficult. Metatarsals I-IV are rod-like. The metatarsal I is stout with a length of 5.7 mm. The metatarsals II–IV are slender and long. Of these, the metatarsal III with a length of 8.1 mm, is the longest. The ansulate bone is large and hook-like. The pedal formula is 2-3-3-3-3. The digits gradually elongate from the digit I to reach the maximum length (16.2 mm) in the digit III, and then digits IV and V become shorter. In contrast, the digit V is 12 mm long and much more slender than other digits. In each digit, the phalanges decrease distally in size. The claw is well developed in anterior four digits. However, the ultimate phalanx is needle-like in the digit V.Sinemys lens, Changmachelys bohlini and Ordosemys liaoxiensis. In contrast to PMOL-AR00007, the juvenile carapace of S. lens is well developed even in the smallest individual (less than 50 mm in carapace length), while the carapace of C. bohlini retains large lateral fontanelles throughout the whole growth series , a circular carapace, lateral carapacial fontanelles, and wide vertebral scales. Few juveniles of sinemydids are known, except for h series . TherefoManchurochelys manchoukuoensis in having a short midline contact of the prefrontals, a postorbital-squamosal separation, a relatively elongated crista supraoccipitalis, and a smaller anterior suprapygal. Also, the specimen is excluded from Sinemys spp. in having a medial prefrontal contact, an elongate crista supraoccipitalis, presence of a cervical scale, a pygal, and eight neurals, and absence of the lateral spine on peripheral 7; Dracochelys bicuspis in having a short medial contact of the prefrontals, a shallow nuchal emargination, eight neurals, a contact of the first peripheral and the first costal, and upturned peripherals. Moreover, the fossil differs from Ordosemys spp., Liaochelys jianchangensis, Xiaochelys ningchengensis, C. bohlini, Kirgizemys spp., Judithemys sukhanovi, and Macrobaena mongolica in not having a postorbital-squamosal contact. In particular, the specimen is distinct from Ordosemys spp. in lacking a preneural plate, but having a long supraoccipital crest, a postorbital-squamosal separation, a broad contact of the first peripheral and the first costal, and a small first suprapygal; L. jianchangensis in having a long supraoccipital crest, and a postorbital-squamosal separation; X. ningchengensis in having a postorbital-squamosal separation, and a small first suprapygal; C. bohlini, Kirgizemys spp., J. sukhanovi, and M. mongolica lacking a broad medial contact of the prefrontals, broad interorbital roof (M. mongolica), and two subequal suprapygals (unknown in C. bohlini). Therefore, it is reasonable to assign the juvenile specimen (PMOL-AR00007) to M. manchoukuoensis.Besides these ontogenetic features, PMOL-AR00007 shows a high similarity with M. manchoukuoensis. Besides the small size, the juvenile features are remarkable in carapace morphology, such as a circular outline, open lateral fontanelles, and wider vertebral scales. In the adult specimen (PMOL-AR00008), the carapace is oval and longer than wide; the lateral fontanelles are completely closed in the well-ossified carapace; among the vertebral scales, vertebrals 2–4 are longer than wide  provides an opportunity to understand the juvenile morphology and ontogeny of han wide . The simhan wide , which imys lens . HoweverSinemys lens, the type species of the family Sinemydidae. All individuals of S. lens come from a single formation and greatly differ in size, and are interpreted to represent an ontogenetic series . The comparative study of M. manchoukuoensis and other sinemydids indicates the trajectory of ontogenetic change appears to be highly diversified in the sinemydids. Of these, the ontogenetic pattern of M. manchoukuoensis is more comparable to that of Sinemys lens, except for earlier occurrence of the well-ossified carapace of the latter. In Changmachelys bohlini and Ordosemys liaoxiensis, the different ontogenetic patterns are also known, such as the circular carapace is relatively independent of ontogenetic stage, and the lateral fontanelles are only closed in adult stage of O. liaoxiensis. In contrast to other taxa poorly-known in ontogenetic change, this diversification of the sinemydids is preliminary in our study and will be highlighted by future studies.Based on the juvenile specimen, our study firstly reveals the juvenile morphology and ontogeny of


This study was conducted in order to understand the implementation quality of the Secondary 1 Program of the Tier 1 Program of the Project P.A.T.H.S.  in the first year of the Full Implementation Phase. Classroom observations of 137 units in 85 schools were conducted under the Co-Walker Scheme. Results showed that the overall level of program adherence was generally high, with an average of 86.57%. Thirteen aspects concerning program delivery were significantly correlated. Multiple regression analyses revealed that (1) overall implementation quality was significantly predicted by interactive delivery method, use of positive and supportive feedback, opportunity for reflection, degree of achievement of the objectives, and lesson preparation; whereas (2) success of implementation was significantly predicted by student interest, interactive delivery method, use of positive and supportive feedback, opportunity for reflection, and degree of achievement of the objectives. In general, the present study suggests that the implementation quality of the Project P.A.T.H.S. is good.


There is an error in affiliation 3 for author Darren P. Baker. Affiliation 3 should be: Sanofi Genzyme, Cambridge, Massachusetts, United States of America.There is an error in the caption for


Babesiosis is a globally important zoonotic disease caused by tick-borne intraerythrocytic protozoan of the genus Babesia (phylum apicomplexa). In China, there are five species that infect cattle buffalo and cause great economic loss, which include Babesia bigemina, B. bovis, B. major, B. ovata, and B. orientalis. Among them, B. orientalis is the most recently identified new Babesia species epidemic in China. This review summarized the work done in the past 33 years to give an overview of what learned about this parasite. This parasitic protozoan was found in 1984 in Central and South China and then named as B. orientalis in 1997 based on its differences in transmitting host, morphology, pathogenicity and characteristics of in vitro cultivation when compared with B. bigemina and B. bovis. It was found that Rhipicephalus haemaphysaloides is the transmitting vector and water buffalo is the only reported host. Phylogenetic analysis based on the 18S rRNA gene also confirmed that B. orientalis is a new species. After species verification, four diagnostic methods including semi-nest PCR, loop-mediated isothermal amplification assay, reverse line blot hybridization assay, and real-time PCR were established for lab and field use purposes. Genomic sequencing was conducted and the complete genomes of mitochondria and apicoplast were annotated. Future work will be focused on developing effective vaccines, identifying drug targets and screening useful drugs for controlling B. orientalis in water buffalo. Babesia orientalis is a tick-borne apicomplexan parasite infecting red blood cells and causing water buffalo babesiosis. It is one of the most important diseases of water buffalo in central and south China, resulting in huge economy loss annually . Phylogenetic analysis based on the nuclear 18S rRNA genes, the amino acid sequences of mitochondrial cox1 and cob genes and heat shock protein 70, confirmed that B. orientalis is a new species which is distinct from B. bigemina and B. bovis , possibly by convenient transportations of water buffalo from south to north because more and more bridges built on the river make the river no longer a natural barrier. In 2011, 14% (16/114) field samples collected north of Yangtze River were positive by real-time PCR   and health water buffalo RBCs (donor of RBC) were needed. According to the record, infected water buffalo blood could be stored for 20 days in 4°C, the parasites still alive for setting up cultivation. Whereas health water buffalo RBC could be only stored for 6 days in 4°C before use ( in 1991 . The medn (MASP) . The parn (MASP) . For setfore use ,b.Babesia and Theileria. Thus, low sensitivity and specificity is one of the most important reasons for developing other testing method. Since 2002, useful serological and molecular methods have been developed for B. orientalis. The first and only serological method was latex agglutination test (LAT) which was established based on in vitro culture excreted antigens of B. orientalis . The sequencing also demonstrated that B. orientalis harbors two extranuclear organelles, mitochondrion and a semi-autonomous plastid–like organelle named apicoplast, which is similar to the other members of apicomplexa parasites. The entire mitochondrial genome is a linear form with 5996 bp in length. It contains three protein-coding genes, cox1, cob, and cox3, in accordance with the mitochondrial genomes of other apicomplexa parasites. Multiple sequence alignment showed that B. orientalis mitochondrial genome is similar to that of the related apicomplexa parasites  pathway. There are seven enzymes work in MEP pathway, including DXS, DXR/IspC, IspD, IspE, IspF, IspG, and IspH, which is similar to previous reported apicomplexan MEP pathway . The results demonstrated that the existence of MEP pathway in B. orientalis and could be a potential drug target for controlling water buffalo babesiosis.The first reported genome of  8.6 Mbp . After tectively . The genarasites . The apiarasites . Further pathway . It was  pathway . The genB. orientalis include three elements, water buffalo, the vector R. haemaphysaloides and the parasites. Control the ticks will be the most efficient way for controlling tick-borne diseases. There are variety of chemicals can reduce the tick populations directly. However, it is impossible to eradicate all the ticks in nature. The other strategy is to control B. orientalis in water buffalo. It was reported that the secretory antigens derived from in vitro cultivation could induce protection to the virulent of B. orientalis, and could be used as a vaccine to prevent the disease  diminazene aceturate is the recommended treatment.Targets for controlling the transmission of  disease , 2002. W disease . AnotherB. orientalis in 1997. Many studies have been done, such as diagnostic methods, vaccine development, genome sequencing, and most importantly, in vitro cultivation was successfully established as it is the basic and core technique for Babesia research. However, little is known about the biology and pathogenesis of the parasite at molecular level. What is the molecular mechanism of R. haemaphysaloides transmits B. orientalis to water buffalo? How does B. orientalis interact with the host, invade red blood cells, grow and proliferate? Several antigens, such as AMA1, RAP1, BoP34, etc., have been identified and presumed to play important roles in B. orientalis invasion of RBC (in vitro cultivated parasites, is expensive and difficult to obtain. Further research will be focused on the understanding of the invasion functions, developing effective vaccines, identifying drug targets and screening useful drugs for controlling B. orientalis in water buffalo, and also the biology and molecular mechanism of vector transmission.It has been more than 30 years since the parasite was discovered in water buffalo in Hubei province, China, and 20 years since it was named as n of RBC ,b. ThereB. orientalis research. QL was a Ph.D. student in the lab from 2001 to 2006. She did the first phylogenetic study and established semi-nested PCR. BY worked in the lab before he retired, he investigated the epidemic areas, studies life cycle and pathogenicity of B. orientalis. All authors read and approved the final manuscript.LH wrote the draft of the manuscript. JZ and MH revised the manuscript. LH, YZ, MH, RF, and JZ are presently working in Parasitology lab, Huazhong Agricultural University, China. They contribute the basic need for 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. The reviewer ME-G and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


Sorex genus are an evolutionarily successful group that includes more than 77 species widely distributed in Eurasia and North America. The genus is one of the rare cases where karyotypic changes reflect well the evolutionary relationships among its species. The taxa showing the greatest variation in karyotype are usually classified into the Sorex araneus group. Its evolution was associated with chromosomal rearrangements, which could have promoted fast diversification of this group into many chromosomal races and species. These processes were additionally complicated by introgressions of mitochondrial DNA, which made the evolutionary history of this group quite complex and difficult to infer. To tackle the problem, we performed multi-method phylogenetic analyses based on mitochondrial cytochrome b that is considered a good molecular marker available for many representatives of Sorex. The results were compared with phylogenies based on chromosomal rearrangement data and put into temporal and spatial context using molecular dating and historical biogeography methods. We complemented the study with the estimation of diversification rates within the S. araneus group as well as comparing the results with paleontological records and climatic oscillations within the last 4 million years. Based on the gathered data, we proposed a hypothetical scenario for the evolution and geographic dispersion of species belonging to the S. araneus group. The shrews began to diversify about 2.7 million years ago in Eurasia and then migrated at least twice to North America. The evolution of shrews was driven by Pleistocene glacial and interglacial cycles, which increased their speciation rate and the emergence of new lineages. The migrations of populations were accompanied by introgressions of mitochondrial DNA into native shrews and occurred at least twice.Shrews of the  Sorex genus, though very common, are unusual because their complex evolution can be deduced from analyses of their karyotypes. This genus has had great evolutionary success and currently includes more than 77 species widely distributed in Eurasia and North America Sorex by e et al. , i.e. 0.e et al. , phylogee et al.  to the me et al. .18O curve [18O records within the same overlapping periods.The maximum clade credibility tree calculated in Beast with associated branching times was used to estimate the diversification rate in R package with LASER 2.4 . To test8O curve , which iSorex: S. araneus Cordon race, S. antinorii, S. coronatus, S. daphaenodon, S. granarius, S. maritimensis, S. samniticus, S. satunini, S. tundrensis from Kemerovo and Tomsk were compared in terms of chromosomal whole-arm rearrangements. The Cordon race was selected as the representative of S. araneus because it shows the most primitive karyotype, which is characterized by polymorphic metacentric fusion j:l and more acrocentric chromosomes than other races [Ten representatives of er races , 35. Theer races , 77–79, er races . We analS. araneus taxa, we carried out likelihood analysis of geographical range evolution (Lagrange) using the dispersal-extinction cladogenesis (DEC) model [S. granarius was placed as the basal lineage to the other members of S. araneus, i.e. before the divergence of S. daphaenodon, as was indicated by the phylogeny based on chromosomal rearrangements. In the model, we determined ten biogeographic regions which are inhabited currently by shrews of the S. araneus group: Eastern, Central and Western America, Northern and Central Asia, Eastern and Western Europe, Caucasus as well as Apennine and Iberian Peninsula. We assumed up to three areas in the ancestral distributions. Range and dispersal constraints were set according to the adjacency of these regions.In order to reconstruct the ancestral geographic distributions of C) model  implemenC) model . The stuS. araneus group, we applied seven approaches based on a popular mitochondrial marker cytochrome b. All methods produced very similar and resolved trees with moderately to very well-supported clades  S. maritimensis and S. arcticus, as well as (2) S. asper and S. tundrensis. Both of them are significantly supported by all methods.To determine phylogenetic relationships within the d clades . The earS. araneus and closely related species, which is very significantly supported by the Bayesian approach and moderately by other methods  and the clade comprising other S. araneus samples (topology t1) were rejected by two and four test, respectively. The location of this haplotype before the divergence of S. coronatus, S. antinorii, S. granarius and S. araneus (topology t5) is less likely and rejected by five tests. Only two positions were not significantly worse than the best tree topology: the grouping of the haplotype B with the whole clade of S. granarius and S. araneus (topology t3), and its clustering with the latter clade, including in addition, S. antinorii (topology t4). Both hypotheses assuming the monophyly of the two S. satunini haplotypes (topologies t6 and t7) were significantly rejected by all tests applied.The placement of haplotype B as a sister clade to the group of S. araneus group was revised several times. S. tundrensis was initially considered a subspecies of S. arcticus and obtained species status after morphometric [S. maritimensis was included within S. arcticus as a subspecies until it was elevated to species status [S. antinorii was also formerly known as the Valais chromosome race of S. araneus and the species rank was obtained after Brunner et al. [S. araneus group. It may also help to recognize the introgression of mtDNA.The taxonomic status of members within the hometric  and genehometric , 23. Sims status , 38. S. r et al.  revision−5, respectively) than the null model assuming that the entire sample comes from a single species with uniform branching. The single GMYC method recognized 17 potential species, the multiple-threshold GMYC method 16 species and the ABGD method found 12 species. The results of various ABGD analyses were consistent and did not depend on an a priori threshold value on the maximal distance.We applied ABGD and GMYC methods . Both GMS. samniticus, S. asper, S. daphaenodon and the S. satunini haplotype A were also recognized as separate clusters. The ABGD and single GMYC methods separated S. arcticus and S. maritimensis but the multiple GMYC considered them as one species, which agrees with the former classification, when S. maritimensis was the subspecies of S. arcticus. The recognition of species within S. araneus and closely related taxa is the most variable. The ABGD produced one cluster including S. coronatus, the haplotype B of S. satunini, S. antinorii, S. granarius and S. araneus. The GMYC methods discriminated S. granarius together with S. araneus Carlit race as one cluster and the other S. araneus as the second one. The S. satunini haplotype B and S. antinorii were also separated by the single GMYC method but the multiple GMYC clustered them into one group. The single GMYC method assigned all samples of S. coronatus to the separate cluster, whereas the multiple GMYC separated the most diverged basal sequence as a singleton.Three outgroup shrews were identified as separate species in all three approaches . Well esS. tundrensis was not recognized as one group but was split into several clusters , a decrease in this rate transpired (to 1.433), which further declined ca. 48 kya (to 0.375)  gives no evidence for the significant slowdown in the diversification. Among tested diversification models, yule4rate turned out the best-fitting with the lowest AIC . The mod375) see . Interes375) see  correspo375) see . In factSorex, we reconstructed their relationships using Bayesian (BA) and maximum parsimony (MP)  and lacks t+u fusion, which is a common feature of S. tundrensis and all other S. araneus representatives. The two S. tundrensis differ by three unique fusions c+l, h+m, and h+p [Using chromosomal whole-arm rearrangement data available for ten representatives of ony (MP)  methods.ntatives , 24–26. -banding . In one ne clade . S. mari and h+p .S. araneus and closely related taxa is occupied by S. granarius, which besides the t+u fissions does not have any other chromosome rearrangements (e.g. a+f fusion) typical of subsequently diverged members. The common grouping of these members is supported by BA and all consensus MP trees. The next separated lineage is S. daphaenodon, which apart from the a+f and t+u fusions, has three other unique ones . All trees with moderate support clustered S. satunini and S. coronatus sharing a unique j+n fusion. The position of S. antinorii is less stable. In BA and two MP trees, S. antinorii groups with the Cordon race. However, in the MP trees assuming ordered characters S. antinorii clusters with the clade S. satunini + S. coronatus. This inconsistency may result from the exclusive presence of l+o fusion in S. satunini and S. coronatus in a monomorphic state and in S. antinorii in a polymorphic state. Simultaneously, the S. araneus, S. antinorii and S. satunini are characterized by b+c fusion, which is absent from S. coronatus.In contrast to the mitochondrial phylogenetic tree, the basal position to the other members of S. araneus lineages were widespread in the whole of Eurasia, from Northern Asia to Western Europe, as these regions were selected as the most probable for the basal node (S. maritimensis and S. arcticus) branches off from this node. A common ancestor of these shrews probably lived in Northern Asia, from where it migrated to Western and Central America across a then-existing land bridge in the place of the current Beringia Strait. The whole Eurasian region was also a diversification centre for further emerged lineages. After the separation of S. tundrensis and S. asper, the ancestors of other current Sorex lineages occupied the region of Europe.The historical biogeography analysis implies that the ancestor(s) of present sal node . The nexS. araneus group, we gathered all the available data and studied it using different methods. The phylogenetic tree based on cytochrome b, provided a well-resolved topology recovered by many methods used. Our results are generally in agreement with other phylogenies based on a smaller number of sequences or simpler approaches [Sorex [In order to reconstruct the evolutionary history within the proaches , 34, 36.s [Sorex , and as S. samniticus to the members of the S. araneus group in the mtDNA tree agrees with its placement in the trees based on karyotypic data. Chromosomes of this species are fully acrocentric, have not been subjected to rearrangements and show substantial homologies to the members of S. araneus group, which implies that S. samniticus represents the ancestral state in relation to the other taxa [S. araneus, S. antinorii and S. coronatus in mandibular characters as well as body mass and measures [The basal position of her taxa , 24–26. her taxa , 37, andmeasures .S. granarius. This species is grouped with significant support with the Carlit race of S. araneus in the mtDNA tree, whereas in the karyotypic trees, it represents a lineage that evolved at the base of the S. araneus group. Assuming that the karyotypic trees represent true relationships between Sorex lineages, the best explanation for this inconsistency is the introgression of mtDNA from the subpopulation of S. araneus to S. granarius [S. granarius from the other members of the S. araneus group in the karyotypic trees well agrees with its clear difference in morphology [S. araneus group and also varies from them in many characteristics of skull and mandible. Discriminant and principal component analyses based on these features clearly separated S. granarius and S. coronatus, which live in sympatry [The subsequently diverging lineages also show the same branching order in the mtDNA and karyotypic trees with the exception of ranarius , 37. Theranarius  and Taberanarius  for a smrphology . It is tsympatry , 85.S. araneus to the Northern Ciscaucasian population of S. satunini [S. satunini sequences are separated in the mtDNA tree into two clades corresponding to the haplotype A and B  [S. satunini is a relatively small population characterized by low polymorphism in microsatellite and morphometric mandibular traits, which is a consequence of its small population size, partial isolation, and genetic drift [Another introgression of mtDNA was suggested to have gone from an ancestor of satunini , 36. In  A and B . The hap A and B . Consequistocene . In agreic drift . The samS. granarius and S. araneus Carlit race were recognized as one cluster by two GMYC methods and the S. satunini haplotype B and S. antinorii were grouped together in the multiple GMYC approach. Interestingly, the ABGD method did not separate S. coronatus, the S. satunini haplotype B, S. antinorii, S. granarius and S. araneus into individual clusters. This may result from the quite recent differentiation of these taxa and might suggest that some hybridization potential between them was retained. Actually, such a phenomenon was reported for S. antinorii and S. araneus [S. coronatus to S. granarius was also proposed [The two cases of introgression are confirmed by the species delimitation analyses.  araneus –44 and tproposed .Sorex araneus taxa  to the Late Pleistocene in Europe and Caucasus Mountains [Sorex form Pliocene of Europe , resembling S. runtonensis [S. subaraneus from the Early Pleistocene deposits  of France or even Pliocene  of Slovakia [S. runtonensis and S. subaraneus are highly polymorphic and could be regarded as a morphometrical continuum of the same species. They may represent ancestors or old remains of the lineages leading to current species. Other detailed studies are required to solve the taxonomic position of these two taxa [Based on available data as well as performed phylogeographic analyses and molecular dating estimates, we decided to propose a hypothetical scenario for the evolution and geographic dispersion of axa Figs  and 9. Mex genus , which iex genus . The emendidates . The oldountains , 89, 90,tonensis . AlternaSlovakia . Under tS. araneus group were probably widespread in the whole of Eurasia. A relict lineage that diverged as the first about 2.7 Mya and survived in the Apennine Peninsula up to the present is represented by S. samniticus (S. runtonensis and S. subaraneus known since the Early (ca. 1.1 Mya) and Late Biharian (ca. 0.75 Mya) of Italy, [S. samniticus lineage implying its continuity in the Apennine Peninsula.The ancestral populations of the mniticus . Unfortuf Italy, , represeS. arcticus and S. maritimensis, lineages emerged also within the Early Pleistocene S. runtonensis or S. subaraneus forms, likely in North-eastern Asia and then migrated across the Beringia Strait into North America about 2.7 Mya  of Colorado and Virginia in USA [The extant  2.7 Mya . This ana in USA , 94—Fig a in USA , which cSorex differentiated into the west and east lineages , with which it shares similar mandibular size and proportions [S. tundrensis is an eurytopic taxon and has diversified into several lineages since ca. 0.870 Mya, which corresponds well with its fossil records known from Moneron Island and dated to the Early Pleistocene (ca. 0.700 Mya) [S. tundrensis several clusters. Although some cytogenetic studies proposed the existence of several species within the Holarctic shrew [S. tundrensis was widely distributed also across Europe during the Last Glacial Maximum [About 2.2 Mya, Eurasian lineages . The extportions . S. tund700 Mya) , 97. The700 Mya) , 39. In ic shrew , additioic shrew , 39. Eco Maximum .S. daphaenodon that spread across Central and Eastern Asia. It must have reached the Far East before 900 kya because its oldest remains are dated to this time in Kolyma Lowland deposits [S. satunini and Western European S. coronatus  deposits from the Baccano maar area in central Italy [S. araneus should be most probably assigned to S. antinorii [S. antinorii lineage. It agrees with our estimation of S. antinorii differentiation dated to about 0.56 Mya.The European populations separated then into the ancestors of  990 kya . Some Mincestors . S. antial areas , 101. Thal Italy . Howeverntinorii . Consequntinorii  can in fS. araneus could occupy Eastern Europe as it is suggested by the basal placement of Russian sequences in the phylogenetic trees Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.


One might say that history repeats itself. The best confirmation of that fact is the 18th International Symposium on Advances in Extraction Technologies—ExTech’2016 and the 22nd International Symposium on Separation Science—ISSS’2016 which took place at the Culture and Congress Center in Toruń, 3–6 July 2016. One more time, over 350 specialists from 41 countries specialising in analytical chemistry, particularly the physicochemical methods of separation, met in the Gothic city of Toruń to summarise their achievements and to present new concepts both in the theory and practice of separation, enrichment, isolation and determination of the entire range of chemical substances. For the first time in history, the two great symposia ExTech and ISSS have been combined as one event. The former pertains to the methods of sample preparation based mainly on the extraction techniques. The latter deals with the physicochemical methods of separation and determination of organic and inorganic substances of different molecular mass and structure, both individual substances and their derivatives occurring in different matrices. The symposia were organised by the Committee of Analytical Chemistry of the Polish Academy of Sciences, Chair of Analytical Chemistry, Faculty of Chemistry of the Nicolaus Copernicus University and the Central European Group for Separation Science (CEGSS). The honorary patronage of this scientific event was assumed by the Minister of Science and Higher Education, Marshal of Kuyavian-Pomeranian Province, Mayor of Toruń, and His Magnificence Rector of Nicolaus Copernicus University in Toruń.The program of the symposium was exceptionally rich. It included 49 plenary and panel lectures as well as 22 oral announcements mostly presented by young scientists. Moreover, during the four subject sessions, there were 258 poster presentations. An integral part of the conference were workshops and the exhibition of 17 manufacturers and distributors of equipment, reagents and accessories. During the symposium, particular attention was drawn to the accomplishments of the founders and pioneers of both extraction science  and chromatography (Prof. Mikhail Tswett). The first session was devoted to the theoretical achievements and fundamental aspects in the field of separation methods. It began with an insightful lecture by Prof. J. Pawliszyn (Canada) concerning the accomplishments and the future of sample preparation technology. Professor P. Jandera (Czech Republic), laureate of the European Tswett-Nerst award, presented the advances which have been made in multi-dimensional separation techniques. A new concept in characterising stationary phases and columns used in the so-called fast chromatography was presented by Prof. A. Felinger (Hungary). Professor O. J. Schmitz (Germany) presented new methodological solutions in the use of coupled separation techniques (LC–MS) for determination of biologically active substances.Similarly, the other sessions brought numerous innovative and interesting solutions and proposals. They concerned such issues as new advances in sample preparation for the needs of biological and environmental analysis including analytics of food and natural products . A particular emphasis was placed on the application of multi-dimensional systems and coupled and combined systems in the determination of biologically active substances on the cellular level (metabolism and omics)  and monitoring changes which take place in the metabolic pathways and biogenic changes .The research paper speeches concerned miniaturisation and robotisation both in the sample preparation and determination of analytes on the sub-ultra traces . These subjects well correspond with the issues connected with developing new generation of materials such as packings, analytical columns and columns used in micro- or nano-scale . The so-called chip laboratory (pill-lab) is a challenge not only for researchers studying miniaturisation and robotisation of the combined process of sample preparation and final determination . This issue was superbly presented in the research paper presented by Prof. Z. Brzóska (Poland), who pointed out the possibilities of using these solutions in modern analytics.The solutions aimed at the application of these achievements for the determination of toxic substances of endo- and exogenic origin or xenobiotics in different matrices, particularly in water, were presented by Prof. H. Lee (Singapore), Prof. M. Bruzzonitti , Prof. D. Knapp (Germany), Dr. V. Comann (Romania) or Dr. M. Gerstiuk (Ukraine), in cosmetics (Prof. M. Llompart from Spain) or monitoring of technological processes (Prof. E. Rosenberg from Austria). All this was perfectly complemented by works devoted to the theoretical considerations containing description of separation mechanisms presented by: Prof. S. Pedersen-Bjergaard from Norway, Prof. I. Malinowska (Poland), Dr. T. Edge (UK), Prof. H. Bagheri (Iran), Dr. J. Weiss from Germany, Dr. D. Berek from Slovakia and Prof. J. Koziel (USA).J. Mazina, A. Tretjakova, P. Saar-Reismaa, J. Gorbatsova, M. Kulp, M. Vaher, E. Erme & M. Kaljurand, Technical University, Tallin (Estonia), poster S1-P71,M. Gładysz, M. Woźniakiewicz, P.M. Nowak & P. Kościelniak, UJ Kraków (Poland), poster S1-P34,T. Hajek, P. Jandera & M. Stankova, University of Pardubice (Czech Rep.), poster S2-P17,M. Skoczylas, Sz. Bocian & B. Buszewski, UMK Toruń (Poland), poster S2-P35,M. Matczuk, J. Legat & M. Jarosz, Politechnika Warszawska (Poland), poster S3-P17,C. Molins-Legua, P. Camping-Falco, Y. Moliner-Martinez, J. Vordu-Andres, J. Pla-Tolos & M. Munnos-Ortuno, University of Valencia (Spain), poster S3-P19,E. Gijonfriddo, E.A. Souza-Silva, S. De Grazia, X. Li & J. Pawliszyn, University of Waterloo (Canada), poster S4-P17,L. Rivoira, S. Studzińska, M. Szultka-Młyńska, M. C. Bruzzonitti, M. Ligor & B. Buszewski, University of Torino /UMK(NCU) Toruń (Poland), poster S4-P33.The presented subject matter was also well suited by short oral announcements made by young researchers (20 papers) and research news presented in the form of posters (258). The latter were also accompanied by short oral presentation in front of a panel of judges headed by Prof. T. Górecki. The jury evaluated the scientific innovation, the form and manner of the presentation. The winners/laureates of the contest were:The material and book prizes were funded by: Springer, Wiley–VCH, Polygen company, Polish Chemical Society and the Committee of Chemical Analytics of the Polish Academy of Sciences.M. Buszewska-Forajta & R. Kaliszan, Gdański Medical University (Poland), OP-7,G. A. Gomez-Rios, N. Reyes-Garces, E. Boyaci & J. Pawliszyn, Waterloo University (Canada), OP-12.The jury also awarded two prizes for the oral presentations of young researchers:The prizes in the form of the covered registration fee at the next conferences of ExTech 2017  and ISSS 2017  were sponsored by the organising committees of these symposia. Moreover, the organising committee of ITP 2017 in Gdańsk awarded a prize to L. Rivoira  by financing the participation in this conference.During ExTech 2016 & ISSS 2016, apart from the up-to-date and abundant scientific program, the organisers provided the participants with an extensive and attractive cultural offer and the presentation of the traditions of the Kuyavian-Pomeranian region and the city of Toruń. The opening concert by Sławek Wiercholski and Nocna Zmiana Bluesa band at the Jordanki Congress and Culture Center does not require any comment. It was an excellent prelude to the working sessions of the symposium. Similarly, the performance given by the Culture Center from Świecie presenting the folk dance, songs and customs of Poland and the region in the setting of the open-air Ethnographic Museum showed the richness and diversity of Polish culture.The show also included a performance by a traditional gypsy band as well as young musical and vocal talents during a common feast, which warmed up the atmosphere of the meeting. Similarly, the participants enthusiastically received the performance of the Toruń singer M. Lubomski and his band given after the official gala dinner at Jordanki Congress and Culture Center. The participants thoroughly enjoyed the delicious dishes served by Gołębiewscy catering company and the excellent music by Rumińscy band. An important element of the program was the post-conference sightseeing tour of Toruń Along the Gothic Trail including a visit to the Live Museum of Gingerbreads. Despite the rainy weather, the participants enjoyed the tour which was confirmed by numerous letters and expressions of thanks for the organisation and the warm familiar atmosphere. This gives us enormous satisfaction of the well-done important tasks that we were entrusted with and makes us committed to live up to the expectations of the future.All this would not have been possible without the invaluable financial support of the government institutions, local and regional authorities and commercial companies (Ministry of Science and Higher Education, Marshal of Kuyavian-Pomeranian region, Mayor of Toruń, Polish Academy of Sciences, companies such as: LECO-Polska, Perlan Technologies, Shim-Pol, AlChem, VWR, TZMO, La Rive and many others. The organisers and the participants express their gratitude for this support on numerous occasions.Prof. Dr. Bogusław Buszewski, DSc.Chairman of the Scientific and Organising CommitteeToruń, 15 July 2016ExTech’2016 & ISSS’2016


Nuttallides truempyi, Oridorsalis umbonatus, Bulimina trinitatensis, the Bulimina simplex group, the Anomalinoides spissiformis group, pleurostomellids, uniserial lagenids, stilostomellids and lenticulinids were ubiquitous during the early Eocene (lower-middle Ypresian). Aragonia aragonensis, the Globocassidulina subglobosa group, the Cibicidoides eocaenus group and polymorphinids became ubiquitous during the middle Ypresian. The most abundant early Ypresian taxa were tolerant to stressed or disturbed environments, either by opportunistic behavior  and/or the ability to calcify in carbonate-corrosive waters (N. truempyi). Nuttallides truempyi, T. selmensis and other buliminids  were markedly abundant during the middle Ypresian. Contrary to the long-lived, highly diverse and equitable Velasco fauna, common and abundant taxa reflect highly perturbed assemblages through the earliest Ypresian, with lower diversity and equitability following the PETM extinction. In contrast, the middle Ypresian assemblages may indicate a recovering fauna, though to some extent persistently disturbed by the lower-amplitude Eocene hyperthermals . We propose the name ‘Walvis Ridge fauna’ for future reference to these Ypresian deep-sea benthic foraminiferal assemblages.The early Eocene greenhouse world was marked by multiple transient hyperthermal events. The most extreme was the Paleocene-Eocene Thermal Maximum , linked to the extinction of the globally recognised deep-sea benthic foraminiferal Velasco fauna, which led to the development of early Eocene assemblages. This turnover has been studied at high resolution, but faunal development into the later early Eocene is poorly documented. There is no widely accepted early Eocene equivalent of the Late Cretaceous-Paleocene Velasco fauna, mainly due to the use of different taxonomic concepts. We compiled Ypresian benthic foraminiferal data from 17 middle bathyal-lower abyssal ocean drilling sites in the Pacific, Atlantic and Indian Oceans, in order to characterise early Eocene deep-sea faunas by comparing assemblages across space, paleodepth and time.  Benthic foraminifera may constitute more than 50% of the total eukaryotic biomass at oceanic depths >1000 m , and areAnomalinoides capitatus and Hanzawaia ammophila; ),Benthic foraminiferal taxa were assigned to microhabitats based on their test morphology: epifaunal species are characterised by milioline coiling, planoconvex and biconvex trochospiral tests, biconvex planispiral, rounded and flat tests, tubular, and leaf-like forms, and infaunal taxa mainly have cylindrical, spherical, ovoid, unilocular globose, rounded planispiral and streptospiral shapes, and also include taxa with a neck e.g., –104). Ho. Ho104])Multivariate analyses, including cluster and non-metric multidimensional scaling (NMDS), have been performed to assess the early Eocene benthic foraminiferal assemblages, using PAST software v. 3.04 . These tBoth analyses were carried out for the intervals corresponding to the lower and middle Ypresian. The lower Ypresian dataset includes 29 species groups from 14 sites , and theThe taxonomy of some of the most common early Eocene species Figs –6, whichBolivinoides and Bolivina , Bolivinoides decoratus  and Bolivinoides cf. decoratus . According to the type description, B. decorata has an elongated test, broad at the apertural end and tapering to rounded point at the aboral extremity, and a surface ornamented with prominent, oblong tubercles arranged in oblique rows [The que rows .Bolivina and Bolivinoides are included in the B. huneri group  [Bolivina huneri (at Site 550), Bolivinoides huneri (at Sites 1262 and 1263) and Bolivinoides crenulata/Bolivina huneri (at Site 401).D’haenens et al.  suggesteNP10-11) , and morB. huneri and B. cf. decoratus groups are somewhat similar, but can be distinguished based on the typical ornamentation of their tests. Although the ornamentation may be slightly blurred in some cases, B. decoratus does not show the pattern of irregularly rounded depressions as in B. crenulata, neither the irregularly anastomosing costae of B. huneri. For this reason, we allocated these species to different groups.The species included in the Virgulina fusiformis by Cushman [This group includes species origninally recognised as  Cushman . The typVirgulina was deemed invalid because it was used to define a genus of a trematode worm, and Fursenkoina was proposed as the new name for this genus [Stainforthia, possibly based on the arrangement of the earlier chambers. Fursenkoina is described as biserial throughout [V. fusiformis. Stainforthia has chambers that are triserially arranged in the early stage, at least in the microspheric generation, followed by a twisted biserial arrangement [Fursenkoina fusiformis and Stainforthia fusiformis were included in this group.The name of the genus is genus . Speciesroughout , but thiangement . SpecimeBulimina elongata and B. thanetensis , however, showed that this twist is not clearly present in some specimens, which thus can be easily confused with B. elongata. Bulimina thanetensis may have become extinct during the early Ypresian, slightly after the start of the PETM , w, wBulimiB. thanetensis  is long and slender, with a markedly twisted test. The first chambers are very small, increasing slowly in size towards the middle part of the test, where they start to increase in size more rapidly, and are more inflated, thus the sutures are also more marked. The holotype slightly differs from deep-sea forms in its more rounded, squat chambers, and less-prominent sutures [B. thanetensis have been found at Maud Rise , whr e.g., , 118), a, aB. thaB. elongata are deposited at the AMNH or NMNH, but we observed diverse Eocene specimens identified as B. elongata . We found a large variety in the width of the test, and in the length-width ratio. Only five specimens from slides CC 59031 and CC 59026 show truly long and slender tests with nearly parallel lateral sides, slightly inflated chambers and slightly depressed sutures. We also checked some hypotypes and plesiotypes from younger time intervals, but these specimens have a more ovate form and flush sutures, in contrast with the Eocene specimens. Nørvang [B. elongata occurs in the microspheric form, and thus these specimens do not correspond to the genera Caucasina as suggested by others [Neither the holotypes nor other primary types of  Nørvang  suggestey others .Bulimina cf. simplex  at Site 401 (B. elongata), but do not have a truly long test. However, they do not have the bulging and protruding chambers which are typical for B. simplex (see B. simplex group).Based on the specimen figured in D’haenens et al. , specimeSite 401  were incBulimina simplex, B. bradburyi and B. tuxpamensis), and without ornamentation or other specific features .According to the original description of otruding . On the chambers , and the sutures . This auB. bradburyi and B. tuxpamensis during the examination of the paratypes of the first species  and the plesiotypes of the second one . However, the plesiotypes of B. simplex  are in general more slender than B. bradburyi and B. tuxpamensis, and show a broad variety of test-shapes. Some have a narrow test at the initial end, with greatest width at the apertural end, whereas others have a more fusiform test. The shape of the chambers varies, from square-rounded and inflated, to longer and more inflated.We noted the above differences between Bulimina prolixa and Pyramidina rudita. Other Ypresian buliminid species such as B. semicostata, B. virginiana, B. farafraensis and B. alazanensis also have bluntly-edged triangular cross-sections, but they can be easily distinguished by their costate ornamentation.The group contains triserial species with blunt-edged triangular cross-section, without costae, e.g., B. prolixa is longer and narrower than P. rudita, and its sutures are slightly depressed. We examined several paratypes of B. prolixa . The specimens have a slender test with triangular cross-section, although they may vary from a typical buliminid shape (largest width close to last chamber) to somewhat fusiform (largest width closer to mid-test).Based on the original description of Cushman and Parker , B. prolP. rudita has a rougher surface but smaller test, with distinct, depressed and sigmoidal sutures [Pseudouvigerina. According to these authors, this genus includes turrilinids with pore mound ornamentation, but their illustrated specimens have a more elevated and pyramidal apertural face than the specimens in Ypresian samples. Moreover, P. cristata [Pseudouvigerina, has an aperture surrounded by a neck , in con Cushman  points od Tappan  mention r et al. . We did u et al. , and theu et al. , and retBulimina semicostata and B. jarvisi, which have a test with coarse perforations and irregularly-shaped, in some cases anastomosing costae  and cotypes of B. semicostata , B. jarvisi is narrower and longer than B. semicostata, and has more inflated chambers and more depressed sutures, in agreement with the original description of Cushman and Parker [B. jarvisi proportionally cover a larger area of the test than in B. semicostata. This ornamentation is finer in B. jarvisi than in B. semicostata [The typical chambers . Based od Parker . The cosicostata .B. asperoaculeata and B. glomarchallengeri. Bulimina asperoaculeata  shows less distinctive costae and deeper sutures than B. jarvisi [B. semicostata may be confused with B. glomarchallengeri, which also slightly resembles the juvenile specimens of B. callahani, but B. glomarchallengeri has more inflated chambers. According to Olshanetskiy [B. glomarchallengeri had its first appearance during the late Eocene, later than the range of B. semicostata.Additional species that may be related to this group include  jarvisi . The micanetskiy , B. glomB. virginiana, B. farafraensis and B. alazanensis) are not included in this group because they can be easily distinguished from B. semicostata or B. jarvisi based on the general morphology of their test. For example, the hypotype of B. virginiana  has a small, narrow test, clearly longer than wide, triangular in cross-section with concave sides and fine costae over the whole test. Some specimens of B. virginiana from ODP Site 1051 and DSDP Site 550 have flat rather than concave sides. Both B. alazanensis and B. farafraensis have a somewhat larger width/length ratio, sometimes with a small spinose projection at the initial end, but the former species has prominent, continuous and more pronounced longitudinal costae, whereas B. farafraensis .Other Ypresian buliminids with costate tests  shows n et al.,  pointed to LeRoy , B. faraPraebulimina reussi (more inflated form) and Bulimina kugleri  was common in the Maastrichtian and became extinct at or close to the Cretaceous/Paleogene boundary globally [Praebulimina and Bulimina, however, have not been clearly described in the literature, and not been observed by all authors.This group has a complex record in the taxonomic literature, and includes buliminids with a typical fusiform shape, smooth test, inflated chambers and depressed sutures, resembling globally , 136, alglobally  reportedB. kugleri is characterised by a marked fusiform shape, with long inflated chambers, and the greatest width in the middle part of the test, with an aperture characterised by a high, arched, curved opening at the base of the inner margin of the last chamber [B. kugleri, but that they are smaller than the type specimens.The species  chamber . The hol chamber . Some of chamber . This auP. reussi rapidly flare from the pointed initial part, and the test has its broadest part in the last whorl [Praebulimina reussi has a subterminal, comma-shaped aperture. We observed three plesiotypes, one of which  has a markedly pointed initial part with small chambers that increase gradually in size, but the chambers rapidly increase in size at approximately one third of the test, resulting in very inflated chambers. The initial part of the other two plesiotypes  is less pointed, the chambers do not abruptly increase in size, and the greatest width occurs in the middle part of the test. We observed that the last chambers of P. reussi are occasionally elongate and inflated, resembling B. kugleri. Similarly, Alegret and Thomas [P. reussi may vary in elongation of the test, ranging from low and globular to elongate and fusiform. In the latter case, these two species may be distinguished because B. kugleri does not show the small, low and compressed early chambers of P. reussi.Chambers of st whorl . Praebuld Thomas  suggesteP. reussi as the species Praebulimina carseyae in their Maastrichtian material [B. ovula Reuss were later renamed P. reussi (Morrow)  e.g., , 30) bec becP. re’Orbigny . There tntricosa  as synoni .P. reussi and B. kugleri, the fusiform buliminid group includes similar Ypresian specimens that have been assigned different names, such as Bulimina sp. 1 (Bulimina kugleri (ovula), Bulimina trihedra, Fursenkoina sp. 2 and Fursenkoina sp. 3.In addition to na sp. 1 , BuliminGlobocassidulina subglobosa  at Site 752 by Nomura [G. subglobosa.The group includes specimens identified as y Nomura , which ay Nomura : Plate 5Pullenia jarvisi, P. subcarinata and P. americana, although their original descriptions indicate they are different species. Pullenia jarvisi has a planispiral test, which is completely involute, biumbilicate and somewhat depressed, with a lobulate outline and about five slightly inflated chambers, separated by depressed sutures. The aperture consists of a low slit extending from one umbilicus to the other, with a strongly convex apertural face [Pullenia subcarinata has a planispiral convex test with a non-lobate outline, slightly keeled, and convex umbilicus [P. americana, which is actually a junior synonym of Pullenia quinqueloba Cushman (not Reuss), has a planispiral, completely involute, much compressed and slightly umbilicate test, peripherally rounded. Pullenia americana has 5 to 6 somewhat inflated chambers, separated by slightly depressed and curved sutures, and an elongate aperture at the base of the apertural face, low at the sides and considerably higher in the middle [P. subcarinata and P. americana have commonly been confused and misidentified in the literature, despite these differences. For example, the SEM images of P. subcarinata in Nomura and Takata [P. jarvisi because of their slightly lobate outline, somewhat depressed sutures and a last whorl with five chambers. Additionally, the upper part of the last chamber is not acute, as in the type figure of P. subcarinata. On the other hand, we expected to observe a higher apertural face than depicted in the SEM image of the specimens named P. americana by Nomura [P. jarvisi. Due to inconsistencies in the identification of these specimens, we decided to include them into a single group, but other flat Pullenia species such as P. quinqueloba (Reuss) and P. salisburyi are excluded as they may be distinguished from this group mostly by their higher apertural face. Likewise, synonyms of the species that we considered misidentified should be detached from this group (e.g. P. quinqueloba).We included into this group the species ral face . Pullenimbilicus . This spe middle . We thind Takata  resembley Nomura , and we P. jarvisi in Arreguín-Rodríguez and Alegret [Pullenia sp. 1 in Alegret and Thomas [Pullenia, because they are not truly planispiral, but should be assigned to Clinapertina subplanispira. Our data from DSDP Site 550 have been modified accordingly in the In addition, we found that many specimens assigned to  Alegret , and Puld Thomas  do not bAbyssamina incisa and A. poagi , with the last chamber more pronounced than in the holotype of A. poagi  but less than in the holotype of A. incisa , and with the degree of depression of the sutures intermediate between both holotypes.According to Schnitker and Tjalsma , A. inciAbyssamina quadrata is somewhat similar to the species of this group, but it can be easily distinguished by its almost planispiral test. Abyssamina quadrata has slightly depressed sutures, as does A. poagi [A. quadrata  than in A. poagi, but it does not look as pronounced as in A. incisa, because all chambers in A. quadrata are inflated. The globular outline of A. quadrata is clearly observed in the images shown by Holbourn et al. [Abyssamina poagi differs from A. quadrata in its longer and narrower chambers, which tend to overlap in the umbilical region, by its more strongly curved umbilical sutures and by its shorter, wider aperture [A. quadrata slightly resembles small and rounded specimens of Pullenia jarvisi, but the aperture in A. quadrata is asymmetrical, whereas it runs from umbo to umbo in P. jarvisi.A. poagi , and hasn et al. . Abyssamaperture . Schnitkaperture  pointed Quadrimorphina profunda also resembles the abyssaminid species, although this species has a low trochospiral test, compressed laterally, with 31/2–41/2 chambers in the last whorl. Its involute side greatly resembles a four-chambered Abyssamina. The main differences with the Abyssamina species are that Q. profunda is trochospiral, with a strongly evolute side (absent in the abyssaminids) that exposes earlier whorls, it has a peripheral aperture with a tendency for extending somewhat onto the evolute side, and it does not show an extension through the umbilical side, as in the abyssaminids. Thus, we did not include it into this group. The figured A. poagi specimen in Holbourn et al. [Q. profunda, in view of the shape of its last chamber extending over the evolute side.n et al.  might, iEilohedra weddellensis and Alabaminella weddellensis are generic assignments used for the species originally described as Eponides weddellensis   . The ori sutures . Our obs sutures  mention A. praeacutus, its periphery is rounded, but the figure of the apertural view in the original description of Vasilenko [A. praeacutus and A. praespissiformis by Nomura and Takata [A. spissiformis. Due to the lack of consistent identification of these species, we included them in one group.According to the type description of asilenko  shows a d Takata  and D’had Takata , respectA. ammonoides, which has been commonly identified in the Cretaceous and Paleocene , whereal . Due tol  is a junior synonym of Anomalinoides rubiginosus [A. rubiginosus ranges from the Upper Cretaceous to the upper Paleocene, and Alegret et al. [Specimens called iginosus , 10, chaiginosus  and Bolliginosus  pointed t et al. , 39 suggt et al.  and Holbt et al. , howeverAnomalinoides capitatus differs from A. rubiginosus in the imperforate periphery of the early chambers, fewer chambers (5–6) in the final whorl, which increase gradually in size, and in the marked sutures varying from thin and depressed in the final chambers to thick and raised in the earlier ones. Van Morkhoven et al. [A. rubiginosus, A. capitatus, A. semicribratus, and A. globosus represent a chronocline from the Late Cretaceous to the present. Thus, the specimens called A. capitatus/danicus by Nomura [n et al.  considery Nomura  may corry Nomura  recognisCibicidoides and Anomalinoides, typically with an unequally biconvex to nearly planoconvex test with numerous  chambers in the last whorl. After careful examination of the data from the study sites, we included Cibicidoides micrus, Anomalinoides cf. acutus, Cibicidoides subcarinatus and C. ungerianus into this group  and two paratypes  of C. micrus, and observed a depressed spiral suture that delimits a circular protuberance formed by a small edge-like extension at the base of the chambers. This feature is more clearly seen in the holotype, but difficult to observe in one of the paratypes, and it was not observed in the specimens identified by Arreguín-Rodríguez and Alegret [According to the description of Bermúdez , C. micr Alegret  at Site C. micrus at DSDP Site 550 and ODP Site 1051 closely resemble those called A. cf. acutus at ODP Sites 1262, 1263 and 690 [Anomalinoides sp. B in Cretaceous-Paleogene sediments at ODP Site 1262 [A. acutus [C. ungerianus and C. subcarinatus respectively, are more similar to C. micrus or A. cf. acutus than to the type figures and descriptions of these taxa. Cibicidoides ungerianus has an acute and carinate periphery, but the specimens illustrated in D’haenens et al. [Cibicidoides subcarinatus has an equally biconvex test with a subcarinate periphery and limbate, comma-shaped sutures, whereas the specimens figured by Nomura and Takata [After reviewing our material, we recognise that specimens called  and 690 , 62 and ite 1262 , but the. acutus . Further. acutus  and Nomu. acutus , we suggs et al.  do not sd Takata  show a mCibicidoides by the elevations of shell material on the dorsal side, and the occurrence of an umbo on the ventral side. Cibicidoides alleni and C. dayi are included in this group.Species of this group are distinguished from other biconvex C. dayi has a strongly depressed spiral suture around a prominent umbo instead of a true elevation of shell material at the base of the chambers on the dorsal side; in addition, its umbo on the ventral side is rather indistinct. In the photographs of type specimens of C. dayi , an umbo on the ventral side may be observed only on specimens with a marked spiral suture, whereas the umbo is indistinct in other specimens.The general morphology of both species is similar, but according to Alegret and Thomas  C. dayi Anomalinoides trinitatensis by Takeda and Kaiho [C. alleni and C. dayi). In the original description of Nuttall [A. trinitatensis also shows a growth of shell material on the dorsal side and a boss on the ventral side, but it has a more planoconvex test. However, specimens in Takeda and Kaiho [A. trinitatensis, in having fewer but more inflated chambers, than those in the type figure, and there is no umbo on the ventral side.Specimens identified as nd Kaiho  are incl Nuttall , A. trinnd Kaiho  differ fCibicidoides micrus also has a depressed spiral suture on the dorsal side, but it is not included into this group because of its smaller size and flatter test, and it was included in the group of small Cibicidoides/Anomalinoides.Cibicidoides mundulus, C. praemundulus, C. pseudoperlucidus and C. proprius  is not a differentiating feature of these two species. However, Alegret and Thomas [C. alleni also differs from C. proprius in the calcite ridge along the spiral side, and in the lack of an acute periphery as in the latter species. For the purpose of this study, we consider these two species  sufficiently distinct to be separated into different groups .The species  Brotzen  described Aubert  suggested Thomas  documentCibicidoides eocaenus in D’haenens et al. [Cibicidoides howelli , deposited at the NMNH, should be included in C. proprius, because they lack the rounded last portion of the test and have an acute periphery. Based on the SEM images in D’haenens et al. [C. eocaenus) should be included in C. proprius, due to their resemblance to the paratypes of C. howelli described by Alegret and Thomas [C. mundulus group.Specimens called s et al.  were re- howelli  Tables. M . AlegreM , its umbilical side is somewhat cone-shaped, and the spiral side is somewhat less convex. The chambers (12–15) are inflated, separated by curved and limbate sutures. Additionally, C. eocaenus has a distinct spiral suture line and frequently a prominent umbilical umbo [C. eocaenus is not included in the Cibicidoides with an umbo group because of its characteristic cone-shaped umbilical side, which makes them easy to distinguish.The species cal umbo . In spitC. eocaenus and C. tuxpamensis may be conspecific. The latter species is very similar to C. perlucidus, characterised by its umbilical plug and acute periphery, but they differ because C. tuxpamensis has a more rounded periphery. According to these authors, there are gradational variations between these two species, which may be considered as ecophenotypic variants and the names perlucidus and tuxpamensis may be used to refer to the periphery of each morphotype. Following this criterion, Arreguín-Rodríguez and Alegret [C. eocaenus perlucidus and C. eocaenus tuxpamensis to designate specimens with an acute and rounded periphery respectively at Site 550 , which has an unequally biconvex test, which is more convex on the ventral side, with an acute and slightly lobulate periphery [The original description of ral side  , but in which the first chambers of the last whorl do not have a clearly trapezoidal shape on the ventral side, as in the type specimens, and with more chambers. These specimens have been called Osangularia sp. 1 at DSDP Site 550 [N. umbonifera .Additionally, we observed specimens that resemble Site 550  and at Obonifera . AccordiT-shaped , but Nutral face . The apeNuttallinella may be distinguished from Nuttallides based on the presence of an umbilicus—rather than a boss—on the ventral side [N. umbonifera, however, have been identified as Nuttallinella ripleyensis or Nuttallinella sp. at ODP Site 1262 [N. umbonifera, and their sutures on the dorsal side are less oblique. Neither of these two species are included in this group because they are not present in the early Eocene.The genus ral side . Cretaceite 1262 . These sNuttallides umbonifera? , Nuttales sp. 2 , Osangulia sp. 1 , Osangularia sp.  and Eponelevatus  should bHanzawaia mantaensis at the AMNH (No. 19954), and were able to identify the holotype shown in the type figure, which is stored in the same slide as several syntypes .In the studied material, this species has been found to be common only at ODP Site 1051. We examined several type specimens of The holotype has a planoconvex test with an acute periphery. The dorsal side is flat and evolute. It shows numerous  narrow chambers in the last whorl, separated by curved, limbate sutures. The ventral side is convex and involute, and the sutures are curved and depressed. According to the original description  and to HH. mantaensis, and we consider the specimens from Site 1051 also as juvenile forms.The specimens identified at ODP Site 1051 do not exactly agree in all features with the holotype, but are more similar to the smaller syntypes in the slide, with a somewhat more convex ventral side, somewhat wider chambers, less raised sutures and a less pronounced umbilicus on the dorsal side, although the proloculus is pronounced . ConsideN. truempyi and Oridorsalis umbonatus, and the B. simplex group, pleurostomellids and uniserial lagenids are ubiquitous , whereas Bulimina trinitatensis and the A. spissiformis group, the P. jarvisi group, lenticulinids and stilostomellids are present in all regions, although not at all sites  are abundant or very abundant, and only 5 ~17%) show a common relative abundance. These 29 taxa are represented in 14 ocean drilling sites from the Pacific, Atlantic and Indian Oceans  and Q. profunda (taxon 1) are abundant at the SE Atlantic sites , but common or present at the seamount sites.Three clusters  are recognised in two separate branches in the dendrogram . One cluQ. profunda and N. truempyi are markedly more abundant than all other species included in the 95% confidence ellipse . Nuttallides truempyi is very abundant in the East Pacific Rise sites, and abundant at Shatsky Rise (Pacific), SE Atlantic and Indian sites. The species distribution across coordinates 2 and 3  and at the East Pacific Rise sites. On the other hand, S. brevispinosa is very abundant at Site 690 (Southern Ocean) and abundant at the other Southern Ocean site, as well as at most Shatsky Rise sites . Inside the confidence ellipse the distribution of the species is mostly associated to the geographic regions where they are abundant, at least considering those species abundant in one region only  than the lower Ypresian, with 42 species or taxa groups. The proportion between abundant (or very abundant) and common species is more balanced than in the lower Ypresian, with 25 taxa (>59%) abundant or very abundant, and 17 (~40%) common. The species ll sites .The dendrogram shows three clusters and two separate branches . One braQ. profunda, pleurostomellids and N. truempyi  than thi; e.g., ), and ali; e.g., ).The fact that the ‘seamount cluster’ includes sites at great distance from each other (Atlantic and Pacific Oceans) suggests that these assemblages may have been controlled by parameters specific to the seamount setting , independent of their geographic location. Such assemblages are strongly dominated by cylindrical infaunal taxa (such as stilostomellids), which seem to prefer relatively high organic carbon flux regularly delivered to the seafloor in non-seamount settings . HoweverEpistominella exigua . For this time interval, we have no data from other North Atlantic sites to test hypotheses about the parameters controlling the benthic assemblages.North Atlantic Site 401  does not cluster with other Atlantic sites, and has an about equal abundance of infaunal and epifaunal taxa , in contd supply , 175), tQuadrimorphina profunda, N. truempyi, T. selmensis and S. brevispinosa are the most notable species, as inferred from the NMDS plots , 36Quadr regions , which ae waters , 39.T. selmensis and S. brevispinosa may have behaved as opportunistic taxa, blooming in response to environmental perturbations associated to the PETM. Both species tolerate stressed/disturbed environments  may have been an opportunistic species, reported from a wide paleodepth range and dependent on food supply, and commonly abundant just after the Paleocene-Eocene extinction [Q. profunda is an opportunistic species that may have become more tolerant to stressed conditions after the PETM, thus accounting for its location relatively close to the other opportunistic species  peaked in abundance together with other opportunistic taxa across the H2 event at DSDP Site 550 [On the other hand, s e.g., , 176) an anT. sels , althouee e.g., , 178). O ellipse . Sprong tinction , 139. Adtinction  suggeste species . FollowiSite 550 .Siphonodosaria subspinosa, Strictocostella pseudoscripta [Assemblages in the middle Ypresian interval have a higher diversity  than those from the lower Ypresian, and higher equitability, i.e., a similar number of abundant and common taxa despite the smaller number of studied sites (9), and the occurrence of several lesser hyperthermal events . The hioscripta ). This soscripta , 62, 63.N. umbonifera group and the G. subglobosa group, but Site 550 does not have abundant B. virginiana, and neither other species such as E. exigua, the B. elongata group or the C. mundulus group, probably due to its greater depth (see below). Despite their seamount setting location, Site 690 is separated from Site 865 in the middle Ypresian  , possibly indicating that Site 690 and Site 550 (which with it clusters) were more susceptible to environmental perturbations during hyperthermal events, with more pronounced effects at great depths (Site 550) and higher latitudes (Site 690). This relation between fluctuations in abundance of opportunistic taxa and hyperthermals is inferred, for example, from major assemblage changes documented during the ETM2 and H2 events (including variations in relative abundances and decreased number of specimens) at Site 550 [As in the lower Ypresian, the sites generally cluster largely by geographical location rather than bathymetry, with a North Atlantic and an Atlantic cluster, and separate branches for Indian and Pacific Oceans. The exception appears to be the clustering of North Atlantic Site 550 with Southern Ocean Site 690 (high latitude cluster), whereas Sites 550 and 401 are clustering far apart, though they are in relatively close geographic proximity  . Site 69Site 550 . FurtherThe middle Ypresian assemblage in the Indian Ocean differs from the other assemblages, as in the lower Ypresian, possibly because the site is much shallower , with a higher food supply and more stable seawater carbonate saturation.B. virginiana, N. truempyi, T. selmensis and the B. cf. decoratus group, with the first species showing palaeogeographic and paleobathymetric restrictions. Bulimina virginiana has been documented from shallower depths along the Northeast coast of the United States , a, aB. virB. virginiana might indicate higher food supply at North Atlantic Sites 401 and 1051, considering that it is a buliminid taxon that lived at shallower, more eutrophic depths. This interpretation is supported by the opposite location of N. truempyi and B. virginiana in the NMDS plot .Abundant MDS plot , which mnditions , 182. Th species , becauses , which s , but its abundance in assemblages with abundant oligotrophic taxa at Site 550 [B. crenulata and B. huneri (which resemble B. decoratus) might indicate a lower food flux than other buliminids. The proximity with the opportunistic T. selmensis [B. cf. decoratus group might have had an opportunistic behavior. These taxa, like T. selmensis, are biserial with an ornamented wall.The Site 550  suggestsSite 550 , 185 argelmensis  in the Nsensu lato , in the uniserial groups , or the pleurostomellids. The dominance of infaunal groups, which has been linked to some combination of high food—low oxygen conditions  inX model, , 187), bX model, , 189). OX model, , carbonaX model, , or chanX model, , 185).Despite these complexities, food supply to the deep sea overall is expected to have been lower in warmer than in cooler periods of Earth history , due toThe high abundance of infaunal taxa in the warm Paleogene oceans has been linked to lower oxygen conditions rather than to a high food supply , 191, duIn addition, the occurrence of multiple short-lived hyperthermal events during the early Eocene , associAnother common feature in the assemblages across both time intervals is the fact that neither the species (and/or groups of species) nor the number of taxa with a global distribution  differ significantly: only 4 taxa that were not ubiquitous in the lower Ypresian became ubiquitous in the middle Ypresian, and 1 group from the lower Ypresian was no longer widely distributed in the upper interval. We suggest that the palaeogeographic distribution of benthic assemblages did not change significantly within the early Eocene (lower and middle Ypresian), although we have data on fewer sites for the middle Ypresian interval . Our present dataset indicates that the relative abundance of benthic foraminifera in the Ypresian was linked mainly to geographic regions , and to a lesser extent to specific environmental settings (seamounts). Sites on depth transects  generally group closely together, whereas sites at similar depths in different regions  do not cluster together . This obN. truempyi, abyssaminids and/or buliminids  as some of the most important species. We also confirm the abundance of buliminid taxa . Furthermore, the abyssal Barbados fauna  from the middle Eocene-Oligocene contrasts to both Ypresian assemblages by its decreasing abundance of N. truempyi (which disappeared at the end of the Eocene together with Abyssamina species), lower abundance of buliminids, and dominance of G. subglobosa [Our assessment of benthic foraminifera across the two Ypresian intervals provides new insights into the main structure and composition of early Eocene global assemblages. Most published compilations have mainly focused on the major extinction across the Paleocene-Eocene boundary e.g., , 27, 31), 3127, 3s e.g., , 139). O. ON. truaxa Figs . In addibglobosa . TherefoBenthic foraminiferal assemblages from Walvis Ridge across the lower and middle Ypresian intervals may be considered as representative deep-sea fauna Figs  and 9, rN. truempyi, Q. profunda, T. selmensis, A. aragonensis, O. umbonatus, N. robusta, pleurostomellids, lenticulinids, and those species comprised into the informal taxonomic groups called fusiform buliminids, B. simplex, G. subglobosa, and A. spissiformis. Other common taxa included in this fauna are C. subplanispira, C. complanata, C. inflata, A. quadrata, E. exigua, S. rugosa, B. trinitatensis, uniserial lagenids, stilostomellids, and the species included in some informal groups such as N. umbonifera, N. havanense, A. incisa-poagi, B. cf. decoratus, B. elongata, costate buliminds, F. fusiformis, and S. brevispinosa. Species such as T. brevispira, B. grata, H. ammophila, A. dissonata, polymorphinids, and those included in the informal groups called P. jarvisi, flat Gyroidinoides, C. eocaenus, C. mundulus, small Cibicidoides/Anomalinoides, E. weddellensis, B. beaumonti, and B. huneri are considered as accessory taxa in the Walvis Ridge fauna.Specifically, the most common taxa that characterise the Walvis Ridge fauna include Not all species included into this fauna shown a global distribution at lower bathyal-abyssal paleodepths (1500–3500 m), but similar assemblages can be recognised in other Atlantic regions and in the equatorial Pacific during the lower-middle Ypresian (∽56–50.7 Ma ).Our study synthesizes and evaluates, for the first time, early Eocene (Ypresian) benthic foraminiferal assemblages living in a greenhouse world with superimposed, short-lived warming events, after a significant extinction. Thus, this taxonomic and paleoecologic compilation constitutes a tool for reconciling different taxonomic concepts and provides an analysis of benthic foraminiferal diversity variability in space and time, and recovery and diversification from PETM extinction.Variations in early Eocene benthic foraminiferal assemblages at the studied sites in the Pacific, Atlantic and Indian Oceans are mainly correlated to differences in palaeogeography , and to a lesser degree to paleobathymetry.The lower Ypresian assemblages are characterised by lower diversity and equitability, i.e., high dominance of abundant species (or groups of taxa) that reflect a perturbed fauna, which resulted from the global extinction associated to the PETM. The most abundant species in this interval may have had an opportunistic behavior and were tolerant to disturbed environments. In the middle Ypresian interval, benthic foraminiferal assemblages reflect a recovering fauna, as expressed in higher diversity and increasing numbers of common taxa, and less affected by environmental perturbations despite the repeated occurrence of hyperthermal events of lesser magnitude than the PETM. The increased diversity and equitability resulted from migration from shallower waters, return of Lazarus taxa from refugia, and evolution of new taxa. We propose the name ‘Walvis Ridge fauna’ for future reference to the early Eocene (Ypresian) deep-sea benthic foraminiferal assemblages, which lived in a greenhouse world with superimposed hyperthermal events, during a phase of recovery from the PETM extinction.S1 AppendixMost common benthic foraminifera during the early Eocene and other species mentioned in the text.(DOCX)Click here for additional data file.S1 Fig(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.


A total of 52 schools participated in the Experimental Implementation Phase of the Project P.A.T.H.S. After completion of the Tier 1 Program, 8,057 students responded to a Subjective Outcome Evaluation Form (Form A) to assess their views of the program, instructors, and perceived effectiveness of the program. Based on the schools' evaluation reports, results of secondary data analyses on four open-ended questions showed that: (a) students felt that they had learned things at the personal, interpersonal, familial, and societal levels; (b) they appreciated the program design, instructors' performance, learning process, and program effectiveness; (c) they generally had positive comments on instructors attitude and teaching process; and (d) they made some suggestions on how the program and its implementation could be improved. The present study, based on qualitative data of subjective outcome evaluation, provides additional support for the effectiveness of the Tier 1 Program of the Project P.A.T.H.S. in Hong Kong.