We conduct a cartography of rhodopsin-like non-olfactory G protein-coupled receptors in the Ensembl database. The most recent genomic data  are analyzed through the online interface and receptors mapped on phylogenetic guide trees that were constructed based on a set of ~14.000 amino acid sequences. This snapshot of genomic data suggest vertebrate genomes to harbour 142 clades of GPCRs without human orthologues. Among those, 69 have not to our knowledge been mentioned or studied previously in the literature, of which 28 are distant from existing receptors and likely new orphans. These newly identified receptors are candidates for more focused evolutionary studies such as chromosomal mapping as well for in-depth pharmacological characterization. Interestingly, we also show that 37 of the 72 human orphan (or recently deorphanized) receptors included in this study cluster into nineteen closely related groups, which implies that there are less ligands to be identified than previously anticipated. Altogether, this work has significant implications when discussing nomenclature issues for GPCRs. Their location as a cellular gateway makes them key targets for drug discovery and chemical biology6 connected by loops. In the rhodopsin family, a well conserved disulphide bridge often connects the second extracellular loop and the third transmembrane segment. This architecture is reflected at the sequence level by conserved sets of amino acids that serve structurally or as determinants of signal transduction, e.g. to name a few the E/DRY, CWxP, NPxxY motifs in the transmembrane segments 3, 6 and 7 of the rhodopsin family8. Additional family-specific motifs have been identified for examples in connecting loops9. Within each transmembrane segment, the most conserved amino acid will be referred to here as pivots . These amino acids are used as a basis of the widely used Ballesteros-Weinstein numbering10, where equivalent positions are reported by transmembrane segment number followed by relative distance to pivot, itself assigned the index 50.GPCRs are well known for sharing a three-dimensional architecture characterized by seven transmembrane α-helical segments13. The largest, Rhodopsin, in human ~700 members, subdivides into four branches  and 13 sub-branches11 and has originated through local duplications about 1400–1100 million years ago14. Families are further divided into subtypes15 that arouse, for many, 350–500 million years ago from two rounds of whole genome duplication (2R)22. In ray-finned fish (not in lobe-finned or jawless fishes), a third specific genome duplication took place about ~250 million years ago, leading to fish-specific duplicates25. In tunicates, GPCRs originated before the 2R and after the separation of invertebrates from chordates26. In lamprey, it remains unclear whether the cyclostome-gnathostome split at the origin of jawed vertebrates happened before or after the second round of genome duplication29.The International Union of Basic and Clinical Pharmacology (IUPHAR) is responsible for the international classification of GPCRs and issues regular recommendations. GPCRs have been divided into five main families based on phylogenetic analyses16, many ohnologues are simply equivalent to subtypes). Nonetheless, local duplications and deletions (pseudogenes)30 may make the current day picture complex and, consequently, the expect four/eight ohnologues are most often not seen30. Furthermore, ligand binding preferences do not necessarily indicates the closest possible evolutionary relationships; some receptors have acquired the same ligand specificity several times, for example α1-, α2-, and β- adrenoceptors that are parts of the amine family32, or e.g. the cannabinoid receptors CB1 and CB2, and the recently deorphanized GPR55 that are activated by the same endogenous ligand 2-Arachidonoylglyserol33.As consequence of the origin of vertebrate GPCRs through the 2R/3R, in an ideal scenario, for each ancestral receptor we expect four/eight paralogues .43 twice with a five-year interval: 2012–2013, Ensembl release 67 (referred to as Ensembl.R67); and 2017–2018, Ensembl.R90-Ensembl.R.92. The text is organized according to the Ensembl.R91 trees . Some of the groupings may slightly differ in future releases. Ensembl.R92 (April.2018) became available during the final preparation of this manuscript and was used to solve a few ambiguities, as mentioned in text. The proposed new gene symbols were submitted to HUGO (Human Genome Nomenclature Committee) for initial consideration; for this purpose the suggested genes symbols were reevaluated against the Ensembl.R.94 (December 2018) and Ensembl.R.95 (February 2019) releases. They will be submitted to the other relevant committees on publication . Therefore all gene symbols presented in this manuscript should be regraded as tentative until approved by the relevant committees.This study was conducted based on the Ensembl genomic dataAltogether, in Ensembl.R92 we identified 142 clusters of genes corresponding to non-olfactory rhodopsin-like GPCRs without human orthologues .  We encourage the readers to look at the most recent data and build an informed opinion.44), in particular, for dopamine receptors45.In the AMIN family Fig.  we sugge2,3,4 receptor family, there are two new fish-specific clades found in coelacanth and ray-finned fishes: see ENSDARP00000127653, named here D2L (gene: DRD2L)(not to be confused with the long splice variant of D2); and see ENSAMXP00000016690, named D4L. Previous work characterized the expression in zebrafish of a single D3 and three D2 receptors46, whereas this study suggest seven zebrafish receptors in the D2,3,4 family: D2a, D2b, D2L, D3, D4a, D4b, and D4L. A previously cloned and pharmacologically characterized α2-adrenoceptor D subtype (ADRA2D) subtype was also found47. Near the histamine receptor H2 (HRH2), rooted by two lampreys, a gene cluster from gar/coelacanth/amphibian/sauropsids(7), see ENSPMAP00000006842, was named GPR187.In the dopamine D7B (HTR7B) found in gar/fishes(10)/coelacanth, see ENSLACP00000008875, that has likely been cloned in zebrafish48; and a set of genes in gar/coelacanth/sauropsids(7), see ENSLACP00000011078, that we name 5-HT7C (HTR7C). A cluster of five genes from gar/coelacanth/birds(3), see ENSLOCP00000003684, may be orthologues of the mammalian 5-HT5B. The 5-HT5B is pseudogenic in humans, but well characterized in mice49.We identified at least two unannotated 5-hydroxytryptamine subtypes: 5-HT1,3,4 subtree, a set of genes from gar/fishes(2)/amphibian/sauropsids(7)/mammals, see ENSLOCP00000006664, was named H5 (HRH5). Near the muscarinic cholinergic receptors 4 and 2, a group of ray-finned fish genes (8), see ENSDARP00000128513, was named CHRM4B; these muscarinic receptors have been cloned from zebrafish50.In the histamine H4 and trace amine (TAAR) receptors, a monophyletic group containing fishes(9)/coelacanth/gar/amphibian, see ENSLACP00000014144, was left unnanotated. Near the 5-HT4 subtree, a set containing gar/fishes(10)/coelacanth genes, see ENSLACP00000004821, was named 5-HT4B . The TAAR have been studied in mouse, rat, human and chimpanzee51 and fishes, where their repertoire is substantially larger than in human53. A set of ray-finned fish genes, see ENSLOCP00000022119, may belong to a previously unannotated subtype.Equally distant from 5-HTIn the MECA family Fig.  we ident188, rooted by lamprey and ascidian sequences, see ENSCINP00000025465. The first set contains gar/coelacanth(duplicated)/fishes(9), see ENSLOCP00000022091; and the second includes fishes(2)/coelacanth/amphibian/reptiles(2), see ENSGMOP00000016757. The duplicated coelacanth sequences open the possibility of a third subtype, but not conclusively.Two sets of genes are likely subtypes of a new orphan receptor, GPR119 has a potential subtype named GPR119B, found in gar/sauropsids(5), see ENSLOCP00000002170.The orphan receptor GPR3, see ENSAMXP00000025994. Nearby, another set has genes of gar/fishes(9)/coelacanth/amphibian/sauropsids(3)/marsupial(1)/monotreme(1), see ENSLOCP00000021577, and corresponds to GPR185; this receptor has been cloned in Xenopus laevis . Two novel sphingosine-1-phosphate receptor subtypes have been previously reported in zebrafish55, which should correspond here to a set of fishes(17)/coelacanth/reptile(1) and to S1PR3a in fishes(10)/coelacanth.A cluster of ray-finned fishes (9) genes suggest a new receptor close to GPR56, UV-sensitive photoreceptors57, and melanopsins58. Zebrafish has 10 classical visual photo pigments and 32 non-visual opsins59.In the OPN family Fig. , one new8a, receptors from a set found in gar/fishes(7), see ENSLOCP00000020544, were named OPN8b. They come in addition to the OPN4–9 in zebrafish59.Near OPNIn the PTGER family, one new subtype was identified Fig. .4, a group of fishes(7)/coelacanth/spotted gar, see ENSLACP00000020254, is a probable new subtype, PTGER4D that has been previously characterized in zebrafish60. In addition, a group of gar/fishes(11)/coelacanth genes, rooted by lamprey, see ENSAMXP00000018684, suggests a new subtype PTGER4C.Near PTGERIn the melatonin tree, no new receptors could be identified Fig. .50 (data from R.95).  Another set includes lamprey/gar/fishes(9)/amphibian/sauropsids(2)/platypus, see ENSLOCP00000014362, was named MTNR1al. A third set of fishes (4), see ENSDARP00000070419, was named MTNR1Ba/1Bb. Three MLT subtypes have been previously cloned in zebrafish61.Ambiguities in this tree were lifted in Ensembl.R92. A complete set containing gar/fishes(10)/coelacanth/amphibian(duplicated)/sauropsids(7)/platypus, see ENSLOCP00000018152, clusters with GPRIn the PEP family Fig. , we iden39 and neurotensin receptors (gene: NTSR), rooted by lamprey, a set of gar/fishes(3)/coelacanth/amphibian genes, see ENSXETP00000007421, suggest a new orphan, GPR189.Near GPRB that contains coelacanth/amphibian/reptiles(2), see ENSLACP00000009190. A closely branched set of gar/fishes(9) genes, see ENSLOCP00000011352, may additionally belong to MLNRB.The motilin receptor (gene: MLNR) has one novel subtype, MLNR2; and a set of fishes , see ENSAMXP00000003803, also orthologous to human GHSR. Four goldfish receptors  have been previously suggested63.The growth hormone secretagogue receptor (GHSR) tree includes three groups of fishes: one containing gar/zebrafish, see ENSLOCP00000009072, likely orthologous to human GHSR; one containing gar/fishes(10)/coelacanth, see ENSLOCP00000000036, that was named GHSR1a, THR1b, THR2, THR3) have been previously characterized in medaka fish64 and three cloned in frog65.The thyrotropin-releasing hormone receptors 1–3 (TRHR) are resolved in Ensembl.R92 as one orthologue and two paralogues of the human TRHR receptor. Four receptors . Two sets of fish genes were assigned to NMUR1 (based on Ensembl.R92). A set of sequences found in lamprey (ENSPMAP00000007962) and gar/fishes(10)/coelacanth, see ENSLOCP00000004052, was named NMUR3.The neuromedin U receptors (NMUR) are divided into NMUR1: a gar/fishes(11) set, see ENSLOCP00000019807, named here NPFFR1L2/3; and gar/fishes(3), see ENSLOCP00000018374, named NPFFR1L1. Two reptile genes branch with either of these receptors.Two groups of genes are likely ohnologues of NPFFR2; and another set with gar/amphibian/sauropsids(7)/marsupials(2), see ENSLOCP00000022432, named PRLHR3. The other group is also rooted by lamprey (ENSPMAP00000008908) and comprises a gar/fishes(7)/coelacanth/amphibian/sauropsids(6) set, named PRLHR4; and another gar/fishes(9)/coelacanth set, see ENSLACP00000014694 (removed from Ensembl subsequent releases). Three homologues of the mammalian receptors have been cloned in chicken66, presumably PRLHR2–4.Two groups of two novel prolactin releasing hormone receptors (PRLHR) are found: the first group is rooted by lamprey (ENSPMAP00000011235) and comprised of one set comprising gar/fishes/coelacanth/amphibian/birds(3), see ENSLOCP00000018414, named here PRLHR6, NPY7, and NPY8 have been described previously70.In the neuropeptide Y receptor family, three unannotated receptors NPY2B; one comprised of fishes(4) genes, see ENSDARP00000119491, named AVPR2l; and one with gar/fishes(14)/coelacanth, see ENSLOCP00000013628, named AVPR2C. Presumably AVPR2l and AVPR2C have been previously reported and named respectively V2-like and V2B71.Three clusters of vasopressin (AVPR) receptor genes are found: one with gar/amphibian/sauropsids(6), see ENSLOCP00000001209, named AVPR72 and four zebrafish GNRHR previously cloned73.We found several interesting gonadotropin-releasing hormone receptors (GNRHR) groups, most including lamprey sequences at their root, whose annotation will require further study since Ensembl.R92 has clarified some of the branchings, but not all. The first contains gar/fishes(27)/coelacanth/amphibian, see ENSLOCP00000017682. The second includes fishes(16)/coelacanth/amphibian/sauropsids(6), see ENSLOCP00000010658. The remaining groups are coelacanth/amphibian/reptile(1), reptiles(5), and placental(31)/marsupial(3) mammals(see ENSMEUP00000014678). The evolution of GNRH receptors has been previously studiedB274.A group constituted of gar/fishes(8)/coelacanth/amphibian/sauropsids(7)/platypus, see ENSLOCP00000018439, likely corresponds to a previously reported endothelin receptor, EDNR15l as a new orphan present in many species: gar/fishes(8)/coelacanth/amphibian/sauropsids(6)/mammals(14 genes from rodents/rabbit), see ENSLACP00000018091. Ensembl.R95 reveals a new subtype of GPR83, annotated GPR83L, and Pgr15lto be a mammalian duplicate of GPR165.Ensembl.R92 resolves Pgr75.In CHEM, we report eight novel receptors, of which four are likely orphans, one divided into two subtypes Fig. . The ass4Rs) we suggest a new orphan receptor GPR190, composed of two (or three) subtypes. A set of gar(duplicated)/fishes(16)/coelacanth(duplicated)/sauropsids(7) genes, see ENSLOCP00000021707, was named GPR190A. The second subtype is composed of gar(duplicated)/fishes/marsupials(2), see ENSLOCP00000021705, and named GPR190B. A lonely platypus sequence (ENSOANP00000021820), complete with respect to its GPCR signature, may represent a third subtype. Two sets of genes near LTB4R1/2 (in Ensembl.R92) may be orthologues of existing human receptors (sets removed in E.95). Three LTB4R have been identified in zebrafish76.Near the leukotriene-4 receptors  sequences, see ENSSHAP00000001972, are likely orthologous to the FRP1,2,3 set. GPR32 is likely rooted by seven monotreme and marsupial genes, see ENSOANP00000022386; and the FRP1,2,3-GPR32 group is likely rooted by a sequence from turtle, see ENSPSIP00000009727.The branching near the formyl peptide receptors FRP1 (in Ensembl.R92), a complex cluster contains four sets of coelacanth genes (some duplicated) branching with two sets of fishes and one set (9 duplicates) of amphibian genes, see ENSLACG00000005198. This group was named GPR1B.Near GPR33 and the chemerin chemokine-like receptor 1 (CMKLR1), taking shape in Ensembl.R92, there is a set comprised of fishes(2)/amphibian/reptile(lizard) genes, see ENSXETP00000062533, that was named GPR191. Nearby is another orphan, with several subtypes, named GPR193 and composed of: coelacanth/reptile(lizard), see eg ENSLACP00000014297; gar/fishes, see ENSLOCP00000021780; isolated genes from lizard (ENSACAP00000004236) and coelacanth (ENSLACP00000021116). There is also a large set of gar/fishes  of ambiguous positioning. Nearby, a set of gar/fishes genes, see ENSLOCP00000022270, was named GPR192; GPR192 may be divided into subtypes since it is branched to coelacanth/sauropsid (5) and amphibian/reptiles(5) sets, indicating an internal duplication in tetrapods.Near GPR8, see ENSLOCP00000022371, as well as to a fish-specific clade containing gar/fishes(8), see ENSLOCP00000000990, named CCR8B. A third group comprising gar(duplicated)/fishes(14)coelacanth, see ENSLOCP00000022422, is located at the root of the mammalian CCR1,2,3,4,5 and CX3CR1 and probably correspond to orthologues of CCR2 and CCR5. We also identified already reported fish-specific receptors, CXCR3B characterized in common carp77 (ENSLACP00000016977) and CCR12A,B identified in orange-spotted grouper (see ENSLOCP00000022265)78.Three sets of fish genes were difficult to assign. Two of these sets are resolved in Ensembl.R92 and tentatively assigned to the fishes orthologous to CCR2) has an additional set gar/fishes(6) genes, probable result of a local duplication, see ENSLOCP00000021801. A set of gar/fishes(20)/coelacanth , see ENSLACP00000020198, may be paralogous to the apelin receptor (APLNR). A second group of fishes(11) nearby, see ENSAMXP00000021714, was assigned as APLNR2.The bradykinin receptor B2 (BDKRB3) has orthologues in fish (ENSLOCP00000021654); a paralogous group comprising gar/fishes(18)/coelacanth, see ENSLACP00000009554. Near RXFP4 (that has fish orthologues too) there is evidence of a new subtype comprised of gar/fishes/coelacanth/mammal sequences, see ENSLOCP00000022049), as well as a fish-specific set of duplicates . These new receptors have been named here RXFP3.# pending further studies and probably have been previously described as RXFP3–2 and RXFP3–380.Human relaxin/insulin-like family peptide receptor 3  group named SSTR2B, probable result of an internal duplication, see ENSLOCP00000021860, has been described previously as SSTR682. Nearby, a group of fishes and coelacanth represent a new SSTR subtype, SSTR1B, since SSTR4 has lost its fishes with the exception of coelacanth83.In the somatostatin (SSTR) receptor group, near SSTR2, there is a gar/fish(10)/coelacanth(duplicated) set, see ENSLACP00000004413. Near MCHR1, two new subtypes are suggested: a coelacanth/amphibian/reptile(1) set, see ENSLACP00000017963; and a larger set comprising gar/fishes/coelacanth/sauropsids(5), see ENSLACP00000009737. Mammals have five melanocortin receptors and one or two MCH receptors, zebrafish six melanocortin (extra MC5RB) and three MCH receptors; fugu four MCR (MC3R missing) and two MCHR; Two MCHR receptors have been cloned in goldfish and  four MCHR receptors have been cloned in Xenopus tropicalis84. No annotations are proposed due to the complexity of the Ensembl trees for this subfamily.Branching close to MCHR1), three unassigned sets of genes were detected. The first contains gar/amphibian, see ENSLOCP00000002856, and was named KISS1RA. The second set contains gar/fishes(10)/coelacanth/amphibian, see ENSLOCP00000003036, and was named KISS1RB; this receptor has likely been studied in ray-finned fishes85. A related set contains gar/coelacanth/reptiles(2), see ENSLOCP00000008391, and the tree furthermore includes fragmental sequences at the root of KISS1 (not discussed here).Near the Kisspeptin receptor 1 (KISS1) has two sets of paralogous fish genes (human GALR1 having its own fishes), rooted by a lamprey sequence (ENSPMAP00000000353). A set of gar/fishes(7)/coelacanth(2)/sauropsids(7) genes, see ENSLACP00000010695, was named GALR1B. Near GALR3, a paralogous branch with gar/fishes(10)/coelacanth/amphibian/sauropsids(4), see ENSLOCP00000001326, was named GALR2B. Avians have been reported to have two new types in addition to Galr1 and Galr2: GalR1, GalR1-like, GalR2-like; four galanin receptors have been isolated from European sea bass 87.In Ensembl.R92, the human galanin receptor 1 , and coelacanth, and was named as GPR196.Two new orphans were identified near the cholecystokinin (CCK) A and B receptors (grouped with PEP in Ensembl.R92). One is composed of fishes(8)/coelacanth(2)/sauropsids(6), see ENSLOCP00000021858, and was named GPR2R) subtypes have been named as UTS2R2–582. A set of gar/fishes(6)/coelacanth/amphibian/sauropsids(6), see ENSLACP00000012826, is orthologous to human UTS2R; A set from gar/fishes(9)/coelacanth/amphibian, see ENSLACP00000003013 is a probable new UTS2R subtype.The Ensembl.R95 gene tree suggest two subtypes of a new orphan receptor, GPR194: gar/fishes(2)/sauropsids/marsupials(2)/monotreme, see ENSLOCP00000022204, named GPR194A; and gar/fishes(10)/amphibian/birds(3), see ENSLOCP00000021948, named GPR194B.Previously four urotensin 2 receptor /coelacanth/amphibian/sauropsids(5)/marsupials(2) genes, see ENSLOCP00000018098, corresponds to the previously described RXFP1B79, annotated here RXFP2L . A fish-specific luteinizing hormone receptor (LH), see ENSLOCP00000020088, has also been reported88.The LGRs are rooted by a set of fish genes  likely orthologous to human LGR1; A group of placental and marsupial mammals, see ENSSHAP00000018195, named MRGPRH. Nearby is a cluster of reptile genes, see ENSACAP00000019338, which may be orthologous or may represent a new subtype. MAS receptors have been reported to arise in amphibians89, and the MAS family tree is rooted by a group of amphibians, MAS1L, see ENSXETP00000063951. Three clusters of fish genes were positioned in Ensembl.R91 with MAS/MRGPRD but in the newer Ensembl.R92 are aggregated with GPR1B .The evolution of the MAS1, MAS1 oncogene-like, and MAS-related GPR (MRG) families is complexified by numerous internal duplications. A large group of sauropsid sequences, locally duplicated, could be orthologous to the mammalian MRGPR, see ENSGALP00000047269, named here MRGPRThe PUR family is the most uncharted and we identified 27 new receptors, including 12 orphans Fig. .5, a set of gar(duplicated)/fishes(2)/coelacanth was assigned to a new orphan probably with two subtypes, see ENSLOCP00000021495, named LPAR5A/B. Near PTAFR, a set of genes comprising gar/coelacanth(duplicated)/sauropsids(2)/mammalian , see ENSLOCP00000002041, was named GPR199.Near LPAR4/6, a set with gar/fishes(4)/coelacanth/reptile(1), see ENSLOCP00000021661, was named GPR202. Another orphan is composed two amphibian and two lamprey sequences, see ENSXETP00000036876, named GPR200. Near GPR55, a full set of genes from gar/fishes(2)/coelacanth(duplicated)/sauropsids(5)/monotreme(1), see ENSLOCP00000021497, was named GPR55B.Near LPAR4, GPR68, GPR65 and GPR132 branch together with a new orphan, named GPR201, divided into two subtypes. One of these subtypes contains gar(7 duplicates)/fishes(2)/reptile(1)/marsupials(2), see ENSLACP00000013584; the other contains gar/fishes(10)/coelacanth, see ENSLACP00000020885.The receptors GPR65 (GPR68 in E.95) there is a set of fishes(5)/coelacanth, see ENSLACP00000000626, was named GPR68B. Near GPR132 a set of gar/fishes(10)/coelacanth genes, see ENSLACP00000009633, was assigned to GPR184. Near P2RY10 and GPR174, a group of gar/fishes(3)/coelacanth genes, see ENSLACP00000004667, was named GPR203.Near GPR2RL3 is prone to local duplications, for example in spotted gar (see ENSLOCP00000014196) or coelacanth (see ENSLACP00000003039). Near F2RL3, a group of amphibian/birds(5), see ENSXETP00000005770, was named F2RL4. A set of fishes(18) paralogous to human F2R, see ENSAMXP00000020201, need further consideration for annotation.The coagulation factor II receptor F2RY8, a group constituted of gar/fish/coelacanth/amphibian/reptiles(2), see ENSLOCP00000021876, was named P2RY8B. Nearby a coelacanth duplicate (ENSLACP00000004454) could not be assigned. In P2RY10, a set of receptors resulting from a mammalian-specific internal duplication, see ENSMUSP00000133122, were assigned to P2RY10B.Near P20 we suggest a new orphan, divided into two subtypes: a group of constituted of gar/fishes(6)/reptiles(3)/monotreme(1), see ENSLOCP00000001967, named here GPR207A; and gar/coelacanth(duplicated), see ENSLOCP00000021450, named GPR207B. A set of fishes(21), see ENSLOCP00000021469, is probably orthologous to the mammalian oxoeicosanoid receptor 1 (OXER1); in such case a set of fish/amphibian(2) genes, see ENSLOCP00000020331, is paralogous. The subtypes 1–3 of the hydroxycarboxylic acid receptors HCA, found only in mammals, are closely related and thus the likely result of recent duplications. Two sets of fishes are branched to the mammalian subtypes, of which gar/fishes(8)/coelacanth, see ENSLOCP00000022252, was named HCAR1L.Near GPR2RY13 has an extra set of fishes(3), see ENSLOCP00000002272, indicating an internal duplication. Near GPR34 there are two groups of sequences: one gar/fishes, see ENSLOCP00000021478, named here GPR206; another set with gar/fish/amphibian/sauropsids(5), see ENSLOCP00000016952, was named GPR34B.P205C and GPR205B; the second gar/fishes(9)/coelacanth/amphibian/sauropsids(4), see ENSACAP00000013831, named GPR205A. Near P2RY1 (NPSR1 in R.95) there is a cluster of genes from gar/fishes(23)/coelacanth/amphibian(triplicates)/sauropsids(7), see ENSLOCP00000022457, that we named NPSR1L. Near P2RY6 there are two sets of unannotated genes: a complete group with gar/fishes(3)/coelacanth/reptile(1)/mammals(21), see ENSLOCP00000021492, named here P2RY16; and, distant and rooted by duplicated lampreys (see ENSPMAP00000011278), a cluster comprised of gar/fishes/coelacanth/amphibian, see ENSLOCP00000021662, that we named GPR204. A set of genes from ray-finned fishes(10)/coelacanth(duplicated)/sauropsids(5)/mammals(5), including one monotreme and one marsupial), see ENSLOCP00000021877, was assigned as GPR197. A gar/fishes(8) specific group of genes, see ENSLOCP00000014291, is branched near GPR17, and we named it GPR17B. Another group rooted by lamprey  comprised of gar/fishes(4)/coelacanth, see ENSPMAP00000011192, was named CYSLTR3A. Two other fishes  nearby open the possibility of another subtype CYSLTR3B.Branched distantly from others is a new orphan divided into three sets: two with genes from gar(2)/coelacanth/reptile, see ENSACAP00000008615, named GPR2#(annotation in process). A set of 32 genes from fishes, see ENSLOCP00000006360, is likely belonging to FFAR2# (Ensembl.R92).Free fatty acid receptors (FFAR) have generally undergone many internal duplications. A small gene cluster from coelacanth/amphibian/reptile, see ENSLACP00000002775, is likely a new new subtype FFAR141C. A set with coelacanth/sauropsids(4)/mammals, see ENSLACP00000005080, was named GPR141B. A set of fishes(10)/amphibian, see ENSXETP00000035434, was named GPR198.These two clusters of orphan receptors were initially aggregated with PURIN receptors but form independent groups in Ensembl.R92. A set containing gar/fishes(10)/coelacanth, see ENSLOCP00000014127, was named GPRWe grouped together receptors that we could not link to the main families, and in this group we identify seven novel receptors Fig. . Interes22L, which may divide further into different subtypes.A set from gar/fishes/coelacanth/amphibian/reptile/marsupial, see ENSLOCP00000017943, is a new subtype GPR210A; gar/coelacanth, see ENSLACP00000019189 (not in R.95); gar/fishes(2)/coelacanth, see ENSLACP00000015589, named GPR210B.Three sets of genes indicate an orphan divided into three subtypes: coelacanth/fishes(4), see ENSLACP00000014196, named here GPR90.A new subtype in ray-fin fishes (8) has been previously reported, which may be a fish-specific duplicate (3R)208; a set of fishes(10)/coelacanth/amphibian/sauropsids(7)/marsupial/monotreme genes, see ENSAMXP00000019172, is distant from GPR20’s placental mammals and may represent a new subtype. A group of gar/fishes(15) is also nearby in Ensembl.R92 (annotated lpar5b), see ENSLOCP00000021496.An independent group of fishes(8), see ENSAMXP00000025847, was named GPR151B.A set of fishes(9)/coelacanth/amphibian, see ENSLACP00000009338, was named GPR209. A second spotted gar, see ENSLOCP00000021505, may indicate another subtype.A set of fish(2)/sauropsids(5)/marsupial(1), see ENSLOCP00000021506, was named GPR62. A set coelacanth/ambiphian, see ENSLACP00000015922, is likely paralogous to GPR211.A set of gar/fishes(10)/coelacanth/amphibian/lizard, see ENSLACP00000013616, named GPR1 and CNR1 in MECA that share sequence identity above 90% towards their human orthologue . Conservation is also high in the AMIN family . In the δ-branch conservation is less . In the unassigned receptors . Not surprisingly, GPR85 , an orphan receptor closely related to the amine family and named "Super Conserved Receptor" shows a very high conservation too  . Generally, as expected the more “ancient” branches such as α-branch and β-branch are relatively conserved compared to more recent branches (χ-branch and δ-branch). In the α-branch Fig. , the mosily Fig.  where thuse Fig. . Few recuse Fig. , but thiess Fig. , and theors Fig. , GPER1 i1,2,3 and GPR32,33), and for distant sequences such as lamprey, have been the most rearranged over the different Ensembl releases. The automated rearrangements have been nonetheless parsimonous, ie groups have been reassigned as orthologues rather than paralogues of known receptors.The novel receptors are identified based on sequence data alone. The complete integrity of the sequences, validated by the conserved GPCR motifs, suggest that these are not pseudogenic. The groupings have been highly stable during the last five years, especially inside monophyletic groups, and most likely to remain so. Generally, the branchings in the fast-evolving PUR and CHEM families  and will continue to be improved following especially the sequencing of novel genomes and the increased coverage of existing genomes.This study maps the extent of orthologues of human GPCRs to vertebrate species. Furthermore, it explores the pool of yet-to-be-studied GPCRs, suggesting at minima 69 sets of novel receptors in vertebrates not orthologous to human. The clustering data allows to group orphan receptors and thus probably brings down the number of endogenous ligands yet to be identified. Nineteen of these groups are maintained in Ensembl.R92 and thus worthy of consideration: In MECA, GPR3/6/12; in PEP, GPR83/165, EDNR/GPR37/37L1, GPR39/NTSR; in CHEM, GPR15/25, GPR182/ACK3; in PUR, GPR20/35/55, GPR101/161, P2RY10/GPR174, GPR17/183, GPR87/P2RY14, GPR31/HCAR; and in other families GPR85/27/173, GPR45/63/135, GPR153/162, GPR139/142, GPR21/52, GPR61/62, GPR26/78; for a few of these receptors an endogenous ligand has been suggestedThe Ensembl.R90 includes 89 vertebrate genomes: eleven ray-finned fishes , one lobe-finned fish, one amphibian, five birds, two reptilians, three marsupial, one monotreme and 65 eutherian mammals, as well as early vertebrates  and invertebrates (one insect and one nematode). The most recent release, Ensembl.R92, includes 94 genomes.92). New orphan receptors were named in numerical order starting from 181, using the numbers left empty (for human genes) by the HUGO Gene Nomenclature Committee. New subtypes were identified either numerically or alphabetically.New receptors were identified based on the branching presented on the Ensembl-generated trees. Three genomes are at the cornerstone for assessing a new receptors: coelacanth, frog, and spotted gar, and we separate them (“spotted gar” referred to simply as “gar”) from the other fishes. Spotted gar diverged from the teleost lineage before the 3RThe data used to build the guide trees are based on an automatically extracted and curated set of 14.000 sequences is described as Supporting Information. This earlier data are used in this manuscript only to build guide trees and compute sequence identities in transmembrane regions.Supporting information


These compounds enable the filling of large air voids in a structure, thus increasing compactness. Therefore, FGG is a more effective repair material to stabilize structures and can function as a sealing and insulating layer. This study used FGG as the repair material for concrete with ground-granulated blast-furnace slag (GGBFS) as the main cement material. The bond strength of the repair was discussed from different aspects, including for fly-ash substitution rates of 0%, 10%, 20%, and 30% and for liquid–solid ratios of 0.4 and 0.5. The slant shear test, and the split tensile test were employed in this analysis. Moreover, acoustic emission (AE) and scanning electron microscopy were used to confirm the damage modes and microstructural characteristics of these repairs. The results revealed that when the liquid–solid ratio increased from 0.4 to 0.5, the slant shear strength of the repaired material decreased from 36.9 MPa to 33.8 MPa, and the split tensile strength decreased from 1.97 MPa to 1.87 MPa. The slant shear test and split tensile test demonstrated that the repair material exhibited the highest effectiveness when the fly-ash substitution was 10%, and revealed that the repair angle directly affected the damage modes. The AE technique revealed that the damage behavior pattern of the FGG repair material was similar to that of Portland concrete. The microstructural analysis revealed that the FGG–concrete interphase contained mostly hydration products, and based on energy-dispersive X-ray spectroscopy (EDX), the compactness in the interphase and bond strength increased after the polymerization between the geopolymer and concrete. This indicated that the geopolymer damage mode was highly related to the level of polymerization.Fly ash/ground-granulated blast-furnace slag geopolymer (FGG) contains reaction products with a high volume of Ca, hydrated CaSiO Scanning electron microscopy (SEM) results demonstrated that repair using fly-ash mortar did not exhibit characteristics of calcium hydroxide or ettringite in the interphase, suggesting that fly ashes alleviate unfavorable microstructural characteristics in the interphase. The concrete–FGG interphase comprised compact C–S–H, and its strength increased slowly in the early stage but significantly in the long term. When fly ash was added, the structure exhibited its highest bond strength, and the decreased number of cracks and amount of carbonization caused dry shrinkage [2 /Na2 base activator to find the optimum substitution ratio, the concrete substrate bonded with the geopolymer slurry. Compressive strength tests have shown that repair rates of up to 120% can be achieved. By comparison with Portland cement, it can be proved that the slag/fly ash base polymer slurry has a good potential for further engineering development in the future. To analyze the damage differences between FGG-repaired concrete and common concrete, the acoustic emission (AE) technique can be used as a monitoring tool in destructive tests. The AE technique is widely applied in nondestructive tests of metal materials, nonmetal materials, and engineering structures, in addition to being used to monitor the crack propagation properties in the fracture process of concrete [Repair materials play a crucial role in extending the lifespan and durability of concrete structures to ensure environmental protection, cost reduction, and enhanced safety. During use, repair materials should be able to sustain the stress change induced by changes in volume in addition to chemical and electrochemical factors, thereby preventing stress relaxation or deterioration and providing the structure with longer durability . The inthrinkage ,6,7. Geohrinkage  used GGBhrinkage . Studieshrinkage  and demohrinkage . The subhrinkage  can be ohrinkage  identifihrinkage  used tunhrinkage  used a mhrinkage  used a hhrinkage  using blconcrete ,19. In tconcrete . AccordiThe results of the aforementioned studies have demonstrated the potential of geopolymers to be used as repair materials. This study examined the effectiveness of FGG as a repair material, and various fly-ash substitution rates were used in FGG for these tests. The different combinations were then tested using slant shear and split tensile tests to establish a reference for using FGG as a repair material in the future.The production process of the specimens comprised two stages: the concrete was first produced and left to stand for 28 days and then repaired using FGG. The specimens were produced in molds and cured in water the day after the molds were removed. For the first part of the concrete specimen to be repaired, the model used in the pouring was the model that the institute built according to the specifications, and the changes were made at different angles. In the second stage, the specimen to be repaired was placed. The traditional model was carried out for repair. The compositions and properties of the concrete are displayed in 2/g and a specific gravity of 2.29 g/cm3, meeting CNS 12549 of the Chinese National Standards.The GGBFS materials used in this study were provided by CHC Resources Corporation . The material had a fineness of 4207 cm2/g and a specific gravity of 2.16 g/cm3.The class F fly ashes used in this study were produced by a Taiwan Power Company thermal power plant . The fly ashes exhibited a fineness of 3818 cmThis study used the type Ⅰ cement produced by the Southeast Cement Corporation , and it met the ASTM C150 Portland cement type I quality standard.3. Before the experiment, the sheet-shaped NaOH was mixed with deionized water to prepare a NaOH solution with a concentration of 14 M. The solid NaOH used in this study was produced by the Zhong Ke company , and exhibited a specific gravity of 0.598 g/cmThis study used the number three sodium silicate solution (37 °Bé) provided by Rong Xiang Industrial Co., Ltd. .3 and a water absorption capacity of 1.93%; moreover, the 3/8” rubble exhibited an SSD specific gravity of 2.65 g/cm3, water absorption capacity of 1.76%, and maximum grain size of 19 mm.The study used natural rubble collected from Ligang, Pingtung in Taiwan. The 3/4” (9.6 mm) rubble exhibited a saturated-surface-dry (SSD) specific gravity of 2.71 g/cm3, and a water absorption capacity of 1.93%.The study used natural sand collected from Ligang, Pingtung in Taiwan. The sand exhibited a fineness modulus of 3.06, an SSD specific gravity of 2.65 g/cmThe FGG specimen production process was as follows. First, the GGBFS was dry-mixed with fine aggregate for 30 s, and then the dry mixture was mixed with an alkali activator for 120 s. After the mixture was left to stand for 15 s, the inner surface of the mixing container was scraped to move the mixed material to the center of the mixing container, and the components were subsequently mixed for 90 s. Next, the FGG was placed in the mold and then rammed and knocked to reduce air voids. After this step, the specimen mold was covered with cling film to reduce water loss. The specimen was demolded after it was air-hardened for 24 h, and then the specimen was cured in a water tank until the desired age was reached.2).A cylindrical specimen with a respective diameter and height of φ10 cm × 20 cm was produced based on ASTM C882 , cured iA cylindrical specimen with a diameter and height of φ10 cm × 20 cm was produced based on Alanazi  and ASTMThe AE technique was incorporated in the split tensile test to monitor the loading process of the FGG-repaired concrete specimen. The establishment of the relationship among the data of event, energy, and indirect tension strength facilitated real-time monitoring of the energy release during the fracture process. As a result, the fracture severity could be quantified.The study used ESEM  to observe the FGG–concrete interphase between the original and the new material, which could be used to predict the different factors that influenced the test. − ions in the alkali solution, impeding the formation of C–S–H gels, and decreasing the strength of the repaired material. A similar trend can be observed in The results of the slant shear test after the repair using FGG are presented in Regarding the angles , the sheThe results of the split tensile test after the repair using FGG are presented in When the fly-ash substitution rate was 0%, the repair strength was favorable in the early stage, whereas the rate of strength growth was decreased in the later stage. When the fly-ash substitution rate was 10%, although the rate of strength growth in the first 7 days was slow, it exhibited a higher strength in the later stage. This was because fly-ashes increase the compactness of a specimen and alleviate the dry shrinkage caused by hot fissures. When the fly-ash substitution rate was above 20%, the strength growth of the repaired structure was slow and substantially decreased. This is because fly-ashes exhibit low activity and slow developmental activity.This study incorporated AE in the split tensile strength test to analyze the process from loading to damage, and the results were divided into three stages for analyses. The frontend stage ranged from the beginning to the first noticeable energy peak, the middle stage ranged from the first energy peak to 80% of the maximum load, and the backend stage was after the end of the middle stage. The test was used to record the energy release during the damage and was one of the methods used to test damage modes. The results are presented in Using the same analysis methods applied by Geng et al. , the preThis study employed a repair angle of 45° and a slant shear test to determine the damage behavior of FGG. The results of the study conducted by Lin  revealed4+ and Al3+, the bond mechanism reacted with the Ca(OH)2 on the surface of the concrete. The increase in Ca2+ counterbalanced the negative charge of Al3+, thus resulting in an increase in hydration products in the FGG–concrete interphase and an increase in bond strength.This study used SEM to observe the interface between the concrete and the FGG, that is, the FGG–concrete interphase, as shown in 1.The slant shear strength and split tensile strength results indicated that the bond strength of the FGG repair material decreased with an increase in the liquid–solid ratio, and a favorable repair effectiveness could be achieved when the fly-ash substitution rate was 10%.2.The damage modes revealed that greater cracking noise was produced when the geopolymers were damaged, and these polymers became sheet-shaped after damage was inflicted; therefore, FGG has low brittleness and fatigue resistance.3.This study incorporated the AE technique into monitoring the damage behavior of the repaired structure, and demonstrated that a higher bond strength produced a higher energy activity in the middle stage, confirming that AE energy is correlated with bond strength.4.Using SEM to observe the microstructural characteristics of this structure, this study revealed that the hydration products in the interfacial interphase were more noticeable when the fly-ash substitution rate was 10% compared with when it was 30%, revealing that the influence of bond strength was highly related to the hydration products present in the interfacial interphase.5.FGG through the slant shear test, split tensile test found that the L/S increased bond strength will drop, and fly-ash amount of 10% can get better repair effect.6.From the failure mode, it is known that there is a large burst of sound when the geopolymer is destroyed, and the failure form is tile-like, so the FGG has the characteristics of low brittleness and poor fatigue resistance.7.We introduced AE to monitor the damage behavior. When the bond strength was better, the energy of the middle part of the sound signal was obviously more active, and it was found that the acoustic energy had a correlation with the bond strength.8.It is found that the influence of bond strength is closely related to the product of the interface interphase when the hydration product of the interfacial interphase is more obvious when the fly-ash amount is 10%.


Cichorium intybus) is a multi-purpose crop and one of the most promising bioactive forages in temperate regions, and numerous in vivo trials have explored its potential against parasitic nematodes in livestock. However, it is unclear whether chicory can induce a direct and broad activity against various GI parasites in different livestock species, and the levels of chicory in the diet that are required to exert an efficient antiparasitic effect. Moreover, the mechanisms leading to the reported parasiticidal activity of chicory are still largely unknown, and its bioactive phytochemicals have only recently been investigated. In this review, we summarise the progress in the study of the antiparasitic activity of chicory and its natural bioactive compounds against GI parasites in livestock, through examination of the published literature. The available evidence indicates that feeding chicory can reduce faecal egg counts and/or worm burdens of abomasal nematodes, but not infections with intestinal worms, in ruminants. Highly chicory-rich diets (≥ 70% of chicory dry matter in the diet) may be necessary to directly affect abomasal parasitism. Chicory is known to synthesise several bioactive compounds with potential antiparasitic activity, but most research has been devoted to the role of sesquiterpene lactones (SL). Recent in vitro studies have confirmed direct and potent activity of SL-rich extracts from chicory against different GI helminths of livestock. Chicory SL have also been reported to exhibit antimalarial properties and its potential antiprotozoal activity in livestock remains to be evaluated. Furthermore, the detailed identification of the main antiparasitic metabolites of chicory and their pharmacokinetics need further confirmation. Research gaps and perspectives on the potential use of chicory as a nutraceutical forage and a source of bioactive compounds for parasite control in livestock are discussed.Increasing drug resistance in gastrointestinal (GI) parasites of livestock and concerns about chemical residues in animal products and the environment are driving the development of alternative control strategies that are less reliant on the use of synthetic drugs. An increasingly investigated approach is the use of bioactive forages with antiparasitic properties as part of the animal’s diet  or as potential sources of novel, natural parasiticides. Chicory ( Infections with gastrointestinal (GI) parasites are ubiquitous in grazing livestock worldwide, inducing subclinical and clinical diseases that can markedly impair animal health, food production and agricultural economies –3. In ruin vitro and in vivo activity under experimental and farming conditions   , 20. Howprograms . In addiIn this review, we summarise the progress in the study of the antiparasitic activity of chicory and its natural bioactive compounds against GI parasites in livestock, through examination of the published literature. In addition, we identify gaps for future research to advance our understanding on the dietary modulation of parasitism using chicory as a model plant, and perspectives on the practical use of chicory as a nutraceutical forage and a potential source of novel plant compounds with broad antiparasitic activity.Chicory is a perennial, deep-rooting herb that can be found as a wild plant in natural grasslands and roadsides and as cultivated varieties in most temperate areas of the world, including Northern Europe , 31. In in vivo effects of dietary forage chicory against GI nematode infections in ruminants. Table Haemonchus contortus and Teladorsagia circumcincta in sheep and Ostertagia ostertagi in cattle), but not infections with intestinal worms in ruminants [T. circumcincta [O. ostertagi [H. contortus FEC [Over the last two decades, several peer-reviewed studies have explored the uminants –56. Secoumcincta , 52 and rtus FEC . The lowrtus FEC , whereasrtus FEC , 58.TablT. circumcincta and Trichostrongylus colubriformis. Subsequently, Scales et al. [Teladorsagia spp. but not intestinal Trichostrongylus spp. in lambs grazing forage chicory, as compared to lambs on control ryegrass (Lolium perenne), cooksfoot (Dactylis glomerata) or tall fescue (Festuca arundinacea). Later studies were in agreement with these findings; e.g. Marley et al. [O. ostertagi/Cooperia spp. third-stage larvae (L3) proportions in faecal cultures, serum pepsinogen or O. ostertagi-antibodies levels, when compared to infected controls grazing ryegrass. A recent study described two independent trials exploring the anthelmintic effects of chicory-rich diets (≥ 70% forage chicory in the DM intake) against experimental infections with GI nematodes in cattle [O. ostertagi and Cooperia oncophora and fed chicory silage diets had a largely similar FEC pattern throughout the trial, in comparison with infected control calves fed a balanced protein/energy diet. However, post-mortem nematode recovery revealed that calves fed chicory silage had significantly lower worm burdens of O. ostertagi (geometric mean reduction of 60%) in comparison with control animals. In contrast, adult counts of C. oncophora in the small intestine were not statistically different between groups. In a second (grazing) experiment, calves mono-infected with O. ostertagi and grazed without possibilities of reinfections on a pure-chicory field (90% chicory DM in the sward), and later supplemented with chicory silage, had similar FEC in comparison with animals grazing a ryegrass/clover pasture until Day 20 post-infection. However, from Day 22 post-infection onwards a rapid and significant reduction in faecal egg excretion was observed in chicory-fed calves, with a 65% lower mean FEC compared with control animals at the end of the trial. Post-mortem worm recovery revealed that calves fed chicory had highly significant reductions in the adult worm burdens of O. ostertagi (geometric mean reduction of 66%) in comparison with control animals [Grazing trials in New Zealand were the first to explore the anthelmintic effects of chicory in livestock , 59, 60.s et al.  reportedy et al.  reportedy et al. –53. In cy et al.  reportedn cattle . In the  animals . These rvs 11.3% in clover-grass) [Furthermore, some evidence of potential indirect anthelmintic effects of chicory has been described , 62. Tzar-grass) . Athanasr-grass)  or by inr-grass) –65. HowePlasmodium falciparum [Oesophagostomum spp. and reduced larval counts of Ascaris suum [Chicory has previously been referred to as a “tanniniferous forage”, although only low levels of condensed tannins (≤ 0.8% in DM) are commonly detected in chicory leaves , 66, 67.lciparum , as welllciparum  and antilciparum  propertilciparum –84. Chiclciparum , 82, 85.ris suum , 87. Theris suum , 89. ThuArtemisia annua  and artemisinin-derivatives  are currently the front-line antimalarial drugs worldwide [Cichorieae tribe (including chicory) around 360 SL have been reported, all part of the groups of guaianolides (~243 SL), eudesmanolides (~73 SL) or germacranolides (~44 SL) [The SL are a group of extremely diverse natural terpenoids, including around 5000 different molecules, which are mainly found in Asteraceae plants . The SL orldwide , 97. Natorldwide , 98. In (~44 SL) . In chicnolides ~43 SL, eu(~44 SL) , 99–102.(~44 SL) , but notDictyocaulus spp. first-stage larvae (L1) incubated in rumen and abomasal fluid from deer grazing chicory. The authors attributed the observed effects to chicory SL in the digestive fluids, although no detection of SL was reported [in vitro effects of water/ethyl acetate crude SL-extracts from chicory roots on the motility of Dictyocaulus spp. L1 and L3 and of GI nematode L3 from deer. However, no chemical identification of SL in the tested chicory extracts was performed [in vitro assays and high-throughput chemical profiling of the tested chicory extracts. Foster et al. [H. contortus by SL-rich extracts from two forage chicory cultivars (“Grasslands Puna” and “Forage Feast”). The authors identified the main guaianolide SL of chicory in the tested extracts  and observed an increased ovicidal activity of the Grasslands Puna-extract, which correlated with its higher content of 8-DOL [in vitro activity of SL-containing chicory extracts against parasitic stages of cattle nematodes (adult O. ostertagi and C. oncophora), which are expected to be main targets of dietary SL in the host [in vitro against the phylogenetically distinct pig nematodes A. suum and Oesophagostomum dentatum, observing also a potent activity on these helminths [50 values obtained in in vitro assays with parasitic stages of cattle and pig GI nematodes exposed to similar chicory SL-extracts. These results indicate that chicory SL-extracts have potent and broad-spectrum activity against parasitic stages of GI nematodes.The anthelmintic potential of chicory SL was first suggested by Schreurs et al.  in a stureported . Furtherreported  describeerformed . Stronger et al.  describeof 8-DOL . Recent the host , 106. Thelminths . Chemicaelminths . Intereselminths , 107. Tain vivo experiments in which chicory diets (with SL profiling) induced a reduction in adult O. ostertagi counts in cattle, the estimated daily consumption of total chicory SL was 27.3 g SL/animal  and 128.3 g SL/animal , equivalent to 222 and 761 mg total SL/kg of body weight in animals from Exp. 1 and 2, respectively [in vivo did not describe the content of SL in the diets, although it is expected that the concentrations of total and individual chicory SL vary depending on cultivar, crop age, season of the year and cultivation site [Aucklandia lappa  [in vitro and did not detect metabolisation of artemisinin to DHA by the rumen microbiota. They also reported a good artemisinin stability at pH 3.0 and 6.8, with a tendency of artemisinin to bind to rumen content and poor solubility in rumen fluid medium [Matricaria recutita) and other Asteraceae plants, is partly metabolised into the anti-inflammatory compound chamazulene carboxylic acid  in artificial gastric fluid, but not in artificial intestinal fluid [Based on two ectively . Howeverion site , 108. Neion site . Yet, thion site , has beeion site –112. Pooion site . Similarlide/kg) . Previoulide/kg) . The samd medium . Similard medium  reportedal fluid . The met2) group attached to the γ-lactone in the SL molecule  at 20% of DM intake, induced a significant reduction in H. contortus worm burdens and FEC [H. contortus in small ruminants. Furthermore, all the available in vivo studies to date testing the antiparasitic activity of forage chicory diets have been performed in ruminants. Promising in vitro activity of SL-extracts of forage chicory against pig nematodes [in vivo, and potentially also explored in other livestock species.The available evidence has confirmed that chicory diets selectively affect abomasal nematodes in infected ruminants, particularly ed lambs . In rela and FEC . Therefoematodes  must be P. falciparum (Honduras-1 strain) in vitro, with total inhibition of parasite growth at 50 and 10 μg/ml of LCP and LAC, respectively [Leishmania spp. has been demonstrated [Eimeria tenella [Neospora caninum intracellular multiplication [in vivo anticoccidial effects of extracts from SL-containing plants  and/or single SL (e.g. artemisinin) have been reported in goats [At present, the antiparasitic activity of chicory has been primarily evaluated in GI helminths and substantial research is needed to explore the potential effects of chicory and its bioactive metabolites against other major GI parasites, such as protozoans. In relation, a previous study confirmed direct antiprotozoal effects of purified SL (LCP and LAC) from chicory roots against the malarial parasite ectively . Besidesnstrated –133. Art tenella  and inhilication . Therefoin goats  and poulin goats –140. TheOstertagia specific antibodies in cattle [in vivo must be elucidated, as previous research has suggested a rapid activity of chicory compounds against parasitic nematodes in vitro [O. ostertagi [in vitro, linked to distinct SL profiles between different varieties [in vivo. Whether sufficiently high concentrations of chicory can be achieved in commercial farming systems needs additional research, though the use of pure chicory swards for livestock nutrition has been explored [In a nutraceutical approach, the existing evidence sustains the use of chicory-rich diets to target adult abomasal nematode infections in ruminants, which are amongst the most pathogenic and economically important parasitic infections in sheep and cattle , 5. Addin cattle , and whein vitro  and a sistertagi . Moreovearieties , 107, reexplored , 47, 144To further validate the practical use of chicory, the economic benefits of including chicory-rich diets in production systems  must be assessed and compared with the extra costs associated with the growth and/or access to chicory. These benefits are expected to vary according to management systems  and in relation to different livestock age groups (e.g. infected with different parasite species). In a nutraceutical approach, and as any other bioactive forage, chicory is also a feed and therefore both its nutritional and antiparasitic value should be evaluated . In addiArtemisia annua and artemisinin [In a future prospect, the comprehensive screening and identification of the main antiparasitic phytochemicals in chicory could help the selection of cultivars with enhanced concentration of these metabolites for use as a nutraceutical forage, or as source of natural parasiticidal compounds. These bioactive metabolites could be selectively isolated to prepare purified extracts that can be used as feed additives for livestock. Such natural antiparasitic feed additives could be given to infected animals in both outdoor (e.g. grazing ruminants) and indoor (e.g. intensive pig) systems. For this purpose, it would be important to define the chicory cultivar(s) and the most suitable part of the plant  for the extraction of active molecules, as well as the standardisation of the extraction technique to efficiently obtain the desired phytochemicals. In addition, lead parasiticidal molecules identified in chicory could be used to develop novel antiparasitic compounds, as previously discovered with emisinin . Once a in vitro. The available evidence has confirmed the in vivo effects of chicory diets against abomasal nematodes in ruminants, and substantial research is needed to elucidate the potential broad antiparasitic activity of chicory against protozoans and in other livestock species. Additional knowledge of chicory’s antiparasitic metabolites and mechanism(s) of action may indicate ways of improving its efficacy, e.g. by selection of cultivars with increased activity and/or the isolation of leading parasiticides. In addition, future research on chicory may also open new paths for exploring other SL-containing plants and improve our basic understanding of the interactions between bioactive phytochemicals, GI parasites and their hosts.Chicory is a bioactive forage with reported antiparasitic activity in ruminants when animals are fed with chicory-rich diets (≥ 70% of chicory DM in the diet). This antiparasitic activity is likely related with its content of bioactive compounds, particularly SL, which have been reported to exert potent effects against helminths and protozoa


QIIME 2 microbiome bioinformatics platform that facilitates access, reproducibility, and interpretation of supervised learning (SL) methods for a broad audience of non-bioinformatics specialists.q2-sample-classifier is a plugin for the  Microbiome studies often aim to predict outcomes or differentiate samples based on their microbial compositions, tasks that can be efficiently performed by SL methods . The goaq2-sample-classifier, a QIIME 2 plugin to support SL tools for pattern recognition in microbiome data. This plugin provides several SL methods, automatic parameter tuning, feature selection, and various learning algorithms. The visualizations generated provide portable, shareable reports, publication-ready figures, and integrated decentralized data provenance. Additionally, integration as a QIIME 2 plugin streamlines data handling and supports the use of multiple user interfaces, including a prototype graphical user interface (q2studio), facilitating its use for non-expert users. The plugin is freely available under the BSD-3-Clause license at https://github.com/qiime2/q2-sample-classifier.We describe The q2-sample-classifier plugin is written in Python 3.5 and employs pandas  and numpThe standard workflow for classification and regression in q2-feature-classifier is shown in RFECV to select the features that maximize predictive accuracy. Hyperparameter tuning is automatically performed using a cross-validated randomized parameter grid search via scikit-learn’s RandomizedSearchCV to find hyperparameter permutations (within a sensible range) that maximize accuracy.The user can enable automatic feature selection and hyperparameter tuning, and can select the number of cross-validations to perform for each (default = 5). Feature selection is performed using cross-validated recursive feature elimination via scikit-learn’s The following scikit-learn  SL estim


The proposed method of preparation of concrete mix using only pre-soaked aggregate (with no extra water) proved to be feasible.The presented research program is focused on the design of a structural lightweight fiber-reinforced concrete harnessing an internal curing process. Pre-soaked waste red ceramic fine aggregate and pre-soaked artificial clay expanded coarse aggregate were utilized for the creation of the mix. Copper-coated steel fiber was added to the mix by volume in amounts of 0.0%, 0.5%, 1.0%, and 1.5%. Test specimens in forms of cubes, cylinders, and beams were tested to specify the concrete characteristics. Such properties as consistency, compressive strength, splitting tensile strength, static and dynamic modulus of elasticity, flexural characteristics, and shear strength were of special interest. The achieved concrete can be classified as LC12/13. A strength class, according to  Usually, the density threshold for concrete to be classified as lightweight is defined as ρ < 1800 kg/m3. The reduction of concrete density can be achieved in different ways: by using porous lightweight aggregates, by introducing a large volume of voids within the concrete mass  and different amount of steel fiber without using additional water. Determined material characteristics of the fresh mix and hardened concrete would be influenced solely by water trapped in both porous aggregates and the process of internal curing [In recent decades, there has been a growing interest in the field of sustainability. In terms of civil engineering, two of the many problems within this field are the massive amounts of demolished brickwork and concrete and the manufacturing of new concrete and bricks, which cause the reduction of natural resources. Therefore, the recycling and reusing waste or unsuitable building materials as a total or partial replacement of traditional concrete ingredients is of highest interest. The ceramic industry produces large amounts of waste each year. The specific characteristics of red ceramic utilized as aggregate could improve the properties of concrete through harnessing the process of internal curing. Lightweight concrete is characterized by density (ρ) significantly lower than the density of ordinary concrete circa 2400 kg/m400 kg/m3concrete ,6. The aThe suitability of lightweight concrete for construction applications is conducted by the needed properties: density, strength, and thermal conductivity. Nevertheless, other properties, such as workability, water absorption, drying shrinkage and moisture movement should also be considered. The porous structure of lightweight aggregates causes high and rapid water absorption during preparation of the fresh mix. Thus, if dry porous aggregate is used for the creation of a mixture, it rapidly absorbs free water, and the workability of the fresh concrete mix is significantly affected . AlthougAn important aspect of the research program is to deepen the knowledge about the usage of waste red ceramic aggregate. The recycling and usage of waste material in the construction field have been discussed by many research groups ,9,10,11,d ≤ 2 mm) were used. Harnessing coarse particles of the waste aggregate for the creation of different concrete mixes was described in multiple previous publications [WRCFA a was solications ,7. One sCommercially available artificial lightweight aggregate was used as coarse aggregate. The expanded clay coarse aggregates ECCA, b are proProperties of both aggregates were tested before the very creation of the concrete mixes. Sieve analysis was conducted according to EN 933-2:1996  using reCopper-coated crimped steel fibers a were utψ and cross-sectional area A were calculated. The straightened length L of every fiber was calculated based on the mean value of the ratio of the wave curve length and the crimped fiber length of 5 fibers. The fiber intrinsic efficiency ratio was determined according to Equation (1) [The fiber cross-section is circular b, so thetion (1) .(1)FIER=The tensile strength of the fiber was tested on 15 randomly chosen fibers. the behavior of fiber during the tensile test is presented in The main purpose of the mix proportioning is to create a formula or a recipe fulfilling a certain requirement . The requirements for the mixture in question were very specific and at the same time quite unconventional: using only pre-soaked lightweight aggregates , good workability of the fresh mix, minimum voids on the surface of samples (closed structure).fV) of concrete 0.0%, 0.5%, 1.0%, and 1.5%. The volumes of added fiber represent the most common additions of steel fiber to concrete. Adding less than 0.5% of fiber does not influence concrete in a noticeable way. On the other hand, using more than 1.5% of fiber would require utilization of an admixture or special mixing and compaction techniques. There are two methods for designing lightweight structural concrete—a weight method and a volumetric method, but it is recommended to use proportions previously established for a similar concrete mixture ,23. ThusAll mixes were prepared in a rotary drum mixer. The mixing procedure lasted 3 min and consisted of 1 min of mixing the aggregate and cement, 1 min of slowly adding the fibers while continuing mixing, and 1 min of mixing all ingredients. After casting the fresh mix into molds, the compaction was performed externally, utilizing a vibrating table for 30 s. Adopted mixing and casting procedures resulted in random 3D spacing of added fiber. The procedures were successfully utilized in previous research programs focused on SFRC based both on natural and waste aggregates.Immediately after the mixing, the slump test (according to EN 12350-2:2009 ) was con3.The specimens were cast in a form of cubes (for compressive and splitting tensile strength), cylinders (for static and dynamic modulus of elasticity) and beams . For the first 24 h, the specimens were kept in their molds tightly covered with polyethylene sheets. Then, the specimens were removed from the molds and put into a water tank. The specimens were cured in the water tank for the next 27 days at a temperature of +21 °C. After the 28-day curing process was finished, all specimens were measured and weighted. The determined density of hardened concretes is presented in 6, the number of pixels representing ECCA in the cross-section is equal to 1.003 × 106, and the amount of pixels representing mortar is equal to 2.035 × 106, which closely corresponds to the ratio of the mix ingredients given in With the help of the open-source graphic software , a simplcf [F at the failure point in kN. The compression strength is given by Equation (2).F—the maximum force; cA—the area.To test the compressive strength fc  three cut,splf [F—the maximum force; L—contact length of the specimen; d—transverse dimension of the specimen.To test the splitting tensile strength ft,spl , three cThe average results of the splitting tensile strength are given in aσ—upper load stress; bσ—lower load stress; εa—mean value of upper load stress strain; εb—mean value of lower load stress strain.Testing the static modulus of elasticity  was execfV = 1.0%, the static modulus of elasticity increased by 9%.The average results of static modulus of elasticity are presented in cuE) was conducted using an ultrasound method. A commercially available ultrasonic testing instrument, usually harnessed for non-destructive testing of concrete strength, was utilized [L—length of the specimen, T—time of the single pulse pass.ρ—the density of the material.The testing of the dynamic modulus of elasticity  is usually assessed by residual flexural tensile strength values that are calculated based on load-crack mouth opening displacement curve or load–deflection curve . A threejF—the load corresponding with CMOD = jCMOD or δ = jδ .Residual flexural strength was calculated using Equation (9).Flexural characteristics of tested beams are shown in R1f and the R3/fR1f ratio (see fib Model Code 2010 [a stands for “pure” softening and d stands for “pure” hardening). Fiber reinforcement can be used also as a substitution of conventional reinforcement to avoid brittleness if the following assumptions are fulfilled [The properties computed based on the bending test and the classification of the SFRC are given in ode 2010 ) was useulfilled .(10)fR3/F—the maximum force, A—an area of the cross-section of the specimen. Shear strength was tested on the half-beam leftovers after conducted flexural tests. The specimens were loaded using shear apparatus continuously, without impact. The values of shear strength were calculated by using Equation (12) .(12)fsheThe average results of the shear strength are shown in 3 and from 14.7 to 17.1 MPa, respectively. These are common values achieved by commercially available lightweight concretes. The uniqueness of the concretes in question is associated with the method of their creation and achieved mechanical characteristics. Creating a lightweight concrete mix is much more difficult than creating an ordinary concrete mix, especially in situ. Most technological problems are usually associated with high water absorption by lightweight aggregate. The aggregate soaks up significant amounts of water, influencing the consistency and workability of the fresh mix. This phenomenon forces producers to use excessive amounts of water or superplasticizer. Both methods have their weaknesses, and they do not guarantee good quality of either the fresh mix or, subsequently, the hardened concrete. In the authors’ opinion, using fully pre-soaked lightweight aggregate makes the process of the creation of lightweight concrete easier and much more predictable. The proposed procedure is feasible to execute even in very basic in situ conditions. Additionally, the created composite benefits from the internal curing process. Due to limited mechanical characteristics, lightweight concretes are commonly used for secondary construction applications. Reinforcing the lightweight concrete in question through the addition of steel fiber enhanced some of its mechanical properties. On the other hand, fiber addition did not influence in a statistically significant way the compressive strength and dynamic modulus of elasticity of the lightweight concrete. This phenomenon is well known, tested, and described in the case of steel-fiber-reinforced concretes based on ordinary aggregate. A similar behavior of fiber-reinforced concretes was noted by multiple researchers [All planned mixes were feasible to create and cast. They were workable and characterized by density and compressive strength ranging from 1366.7 to 1484.9 kg/mearchers ,39 when fV = 1.5%). Due to fiber addition, the lightweight concrete gained quasi-plastic flexural characteristics. According to fib Model Code 2010 [fV = 0.5%, fV = 1.0%, and fV = 1.5%, respectively. In all three cases, the substitution of traditional reinforcement is enabled. The conducted research program proved that pre-soaking lightweight aggregate  enables the reasonably easy production of lightweight steel-fiber-reinforced concrete. This non-conventional concrete is in need of further lab and application tests. It creates a realistic opportunity for harnessing huge volumes of waste red ceramics.Fiber addition influenced splitting tensile strength, static modulus of elasticity, and shear strength by 30%, 17%, and 18%, respectively (for ode 2010 , the tes-It is feasible to create concrete using only pre-soaked porous aggregates and no extra water;-The proposed method of the creation of lightweight concretes with no extra water is easy to execute both in laboratory and in situ;-Achieved density and basic compressive strength enable created concretes to be classified using strength classes according to EN 206-01;-fib Model Code 2010 [Achieved flexural characteristics enabled using ode 2010  strength-All cast SFRC are enabled for reinforcement substitution;-The mechanical properties of cast SFRC enable using them for the chosen structural applications;-Additional tests associated with the durability and thermal conductivity of concretes in question should be conducted.The research program allows drawing the following conclusions:


The use of self-consolidating concrete and advanced pumping system enables efficient construction of super high-rise buildings; however, risks such as clogging or even bursting of pipeline still exist. To better understand the fresh concrete pumping mechanisms in detail, the discrete element method is employed in this paper for the numerical simulation of local pumping problems. By modeling the coarse aggregates as rigid clumps and appropriately defining the contact models, the concrete flow in representative pipeline units is well revealed. Important factors related to the pipe geometry, aggregate geometry and pumping condition were considered during a series of parametric studies. Based on the simulation results, their impact on the local pumping performance is summarized. The present work demonstrates that the discrete element simulation offers a useful way to evaluate the influence of various parameters on the pumpability of fresh concrete. The growing peak of landmark buildings, such as the Shanghai Tower (632 m high), Burj Khalifa (828 m) and the Jeddah Tower (planned to be over 1000 m), is attributable to the continuous theoretical research and technical development particularly in the field of civil engineering. In high- and super high-rise building projects, concrete pumping has become a very important part of construction technology to guarantee the project schedule. The smooth pumping of fresh concrete directly from the ground surface up to several hundreds of meters height demands an optimal combination of material, equipment and process. The concrete material itself should be highly flowable, which is now generally available thanks to the invention of self-consolidating concrete (SCC) in the late 1980s. The pumping apparatus consists mainly of the pipeline connected to the pump. The pipes should be tough enough to sustain the pressure and abrasion, and the pump should provide sufficient power for the concrete to overcome the gravity and friction. Nevertheless, despite the general success in the completed projects, unexpected situations  can still occur in reality, which may significantly influence the project schedule and even cause hazards to the people on site.Substantial understanding of the flow behavior of fresh concrete during pumping will provide a good foundation for the solution of existing and potential problems in construction projects. Although fresh concrete is relatively less studied as compared to concrete materials and structures in hardened state, scientists and engineers have made considerable efforts in terms of experimental tests, theoretical analyses and numerical simulations. From the mechanical point of view, fresh concrete is nothing but a flowable material similar to smog, plasma, petroleum, debris flow, etc., and can be studied using classical or modern methods for fluids .The flowability, more specifically the rheological property, of fresh concrete is dependent on various factors such as the mix design, internal action and environmental condition, and also complex mechanisms between them . Fresh cThe rapid development of the numerical simulation of concrete flow in recent years has brought a variety of theory and modeling techniques  can be treated as continuum at the macroscopic point of view, because the characteristic length of material ingredient is very small as compared to the problem domain. If one focuses on the local behavior of material at a length scale where the microstructure is no longer negligible, discontinuous modeling should be considered . The comConcerning the specific concrete pumping problem, few publications report its numerical simulation. As a first choice, CFD is used to simulate the concrete flow as viscous fluid in the pipe; as a result, the total pressure-flow rate relationship is obtained and compared with a real-size pumping experiment ,43). In This article reports a systematic DEM study of local behavior of fresh concrete during pumping . We invediscrete element method specifically employed in this paper was initially referred to as the distinct element method and was proposed for the study of granular assemblies [The semblies ,29. In tsemblies ,49,50,51particles , and can also be influenced by the changing rate of gap . A contact model consisting of the normal and shear components can be used for both the particle-particle and particle-boundary contacts  and to calculate contact forces contacts a. In theConcrete is a heterogeneous material composed of coarse aggregates, fine aggregates, dry cement, water and other chemical additives. In a DEM simulation of concrete flow, the goal is to capture the motion of aggregates that directly reflects the flow behavior of fresh concrete. In the DEM model, an individual particle represents a specific piece of aggregate. Usually in the model, a particle is simply a ball (a disc in 2D), which allows efficiently determining the geometrical relations between different entities during computation. If more attention is paid to the influence of irregular shape of aggregate, non-spherical particles can be generated via specific algorithms , which are already or potentially in contact with the fresh concrete, are usually modeled as rigid walls, either stationary or kinetic according to the given boundary conditions.Fresh concrete is often treated as single phase non-Newtonian fluid from the macroscopic point of view. In the framework of rheology theory, concrete material parameters such as the yield stress and plastic viscosity can be determined based on specific laboratory tests. These parameters are used as input for a typical CFD simulation of concrete flow ,11,22,57In DEM, the bulk material is described at the meso-scale as a collection of particles. During DEM simulations, a particle itself is merely a rigid body; it is the contact model and model parameters that govern the motion of every individual particle. Before conducting the DEM simulation of concrete flow, the definition of contact model and model parameters is a necessary step. In the literature, different models exist. Some researchers use experimental methods to investigate the aggregate–aggregate and aggregate–boundary contact behaviors, from which new contact laws are proposed ,30,58,59linear parallel bond model, which belongs to the collection of built-in contact models in PFC 5.0 [linear model mainly describes the contact behavior under compression; therefore, it can be used for dry materials. The linear contact bond model has a pair of tensile and shear springs, which, when active, preclude the slip and friction at contact. Other models contain more complex components such as nonlinear spring, nonlinear viscoelastic element, a planar interface, rolling resistance, Kelvin and Maxwell models in series, etc. The fundamental formulations of the linear parallel bond model are briefly provided in the following , and commercial programs including UDEC , EDEM software and PFC (Particle Flow Code). In the present research, simulations were conducted using PFC 3D 5.0 (Particle Flow Code 3D Version 5.0) . We used PFC 5.0 ,65. In fentation ).As illustrated in For the parallel bond, the force and moment are calculated as followsThe maximum normal and shear stresses at the contact are calculated asAs mentioned previously, a DEM model describes the material at the meso-scale; therefore, the defined contact laws and parameters are not equivalent to the conventional macroscopic material property, which can be directly measured with standard experiments. For the determination of mesoscopic model parameter values, a number of methods can be considered. For example, the contact spring stiffness can be derived from the Young’s modulus of material ; the friIn the present work, a “standard” self-consolidating concrete was assumed, without referring to a particular concrete mix. For the DEM modeling of this virtual SCC, the particles included equal volume of four types of clump , and theThe parameter values in In modern construction projects, fresh concrete is usually provided by mixing plants and pumped through pipelines that can reach a total length of several hundred meters. However, the goal of present work was not the DEM simulation of whole pumping system; instead, we limited our scope to the local simulation of concrete flow through representative pipe units, with the focus on the different performance in various pumping cases characterized by different parameter setting in terms of model geometry, material property and pumping condition. As demonstrated below, the simulation of pumping units can already be used to quantitatively evaluate the pumping performance of whole system. Furthermore, the local simulation results reveal how a specific parameter influences the pumping resistance, and thus provide optimization directions for the pumping program. Nevertheless, the realistic situation in a construction site can be very complicated; there are many influencing factors such as the change of concrete workability, environmental conditions, the assembly of pipeline, the actual pressure and velocity at the outlet of pump. It would be a meaningful and challenging topic to identify all the relevant factors, introduce them appropriately into the numerical model and assess their impact on the pumping performance.d; the elbow radius was R. The geometrical model of pipe was built in CAD software and exported as STL-file, which was later imported by the DEM software to form the boundary .As illustrated in In the DEM simulations, the reference concrete model was first adopted, as described in P induced by the gravity, friction and deformation effects.A unit length (1 m) of fresh concrete was modeled. The flow of fresh concrete was driven by a displacement-controlled wall that was perpendicular to and moved along the pipe axis . A “plugv; and the second was the “stable pumping stage” when the wall moved along the pipeline with constant v. Since the DEM essentially deald with dynamics, the stability of numerical scheme was ensured by limiting the time step size The pushing of wall consisted of two stages: a first “acceleration stage” in which the velocity of wall was linearly increased until reaching the nominal pumping rate We started with the situations of concrete pumped through different pipe units. As a reference setup, the following condition was considered: pipe diameter P) vs. displacement (u) relationship from each case is plotted in P was calculated as the reaction force on the wall divided by the cross sectional area of pipe, and the displacement u was the wall displacement along the axis of pipeline. The vertical gray line at displacement The thrusting pressure  reduced the pumping difficulty marginally.  after the concrete is freshly produced at the mixing plant; otherwise, the workability of concrete deteriorates with time due to several reasons such as hydration. Here, we conducted the DEM simulation to test different hydration states, without referring to any exact value in reality. We randomly selected a proportion of particle contacts and assigned higher values to the bond stiffness portions , right. It was interesting to look into the extreme situation of g to 20% . ClearlyThe present paper reports the DEM modeling and simulation of the pumping behavior of self-consolidating concrete at a local level, considering different situations of concrete pushed through a pipeline unit. The flow of concrete is reflected by the motion of particles that represent coarse aggregates, and the mortar phase is physically taken into account as the contact behavior. A series of parametric studies was carried out, with respect to the influence of pipe geometry, material property and pumping condition. The simulation results suggest that the gravitational effect takes a big part of the pumping resistance; an elbow does not cause significant increase of pressure level; smaller pipe diameter or larger aggregate size leads to dramatic pressure growth in a nonlinear manner with increasing slope; the aggregate shape factor or the pumping rate has a linear influence on the pumping resistance; and the hydration of fresh concrete should be controlled. In general, the DEM simulation of concrete flow not only helps to understand the details of pumping behavior, but also inspires new concrete pumping techniques that can be considered to eliminate pumping risks and to confront more challenging circumstances such as record-breaking pumping height in the future.Limitations have also been noticed during the present numerical study, from which subsequent research is being considered. The mesoscopic model of fresh concrete should be improved, including more accurate geometrical representation of aggregates , more so