Phylogenomics of Neogastropoda: the backbone hidden in the bush.

The molluscan order Neogastropoda encompasses over 15,000 almost exclusively marine species playing important roles in benthic communities and in the economies of coastal countries. Neogastropoda underwent intensive cladogenesis in early stages of diversification, generating a 'bush' at the base of their evolutionary tree, that has been hard to resolve even with high throughput molecular data. In the present study to resolve the bush, we use a variety of phylogenetic inference methods and a comprehensive exon capture dataset of 1,817 loci (79.6% data occupancy) comprising 112 taxa of 48 out of 60 Neogastropoda families. Our results show consistent topologies and high support in all analyses at (super)family level, supporting monophyly of Muricoidea, Mitroidea, Conoidea, and, with some reservations, Olivoidea and Buccinoidea. Volutoidea and Turbinelloidea as currently circumscribed are clearly paraphyletic. Despite our analyses consistently resolving most backbone nodes, three prove problematic: First, uncertain placement of Cancellariidae, as the sister group to either a Ficoidea-Tonnoidea clade, or to the rest of Neogastropoda, leaves monophyly of Neogastropoda unresolved. Second, relationships are contradictory at the base of the major 'core Neogastropoda' grouping. Third, coalescence-based analyses reject monophyly of the Buccinoidea in relation to Vasidae. We analysed phylogenetic signal of targeted loci in relation to potential biases, and we propose most probable resolutions in the latter two recalcitrant nodes. The uncertain placement of Cancellariidae may be explained by orthology violations due to differential paralog loss shortly after the whole genome duplication, which should be resolved with a curated set of longer loci.

Coupling DNA barcodes and exon-capture to resolve the phylogeny of Turridae (Gastropoda, Conoidea).

Taxon sampling in most phylogenomic studies is often based on known taxa and/or morphospecies, thus ignoring undescribed diversity and/or cryptic lineages. The family Turridae is a group of venomous snails within the hyperdiverse superfamily Conoidea that includes many undescribed and cryptic species. Therefore 'traditional' taxon sampling could constitute a strong risk of undersampling or oversampling Turridae lineages. To minimize potential biases, we establish a robust sampling strategy, from species delimitation to phylogenomics. More than 3,000 cox-1 "barcode" sequences were used to propose 201 primary species hypotheses, nearly half of them corresponding to species potentially new to science, including several cryptic species. A 110-taxa exon-capture tree, including species representatives of the diversity uncovered with the cox-1 dataset, was build using up to 4,178 loci. Our results show the polyphyly of the genus Gemmula, that is split into up to 10 separate lineages, of which half would not have been detected if the sampling strategy was based only on described species. Our results strongly suggest that the use of blind, exploratory and intensive barcode sampling is necessary to avoid sampling biases in phylogenomic studies.

Collaborative Expression: Transcriptomics of Conus virgo Suggests Contribution of Multiple Secretory Glands to Venom Production.

Venomous marine gastropods of the family Conidae are among the most diversified predators in marine realm-in large due to their complex venoms. Besides being a valuable source of bioactive neuropeptides conotoxins, cone-snails venoms are an excellent model for molecular evolution studies, addressing origin of key innovations. However, these studies are handicapped by scarce current knowledge on the tissues involved in venom production, as it is generally assumed the sole prerogative of the venom gland (VG). The role of other secretory glands that are present in all Conus species (salivary gland, SG) or only in some species (accessory salivary gland, ASG) remains poorly understood. Here, for the first time, we carry out a detailed analysis of the VG, SG, and ASG transcriptomes in the vermivorous Conus virgo. We detect multiple transcripts clusters in both the SG and ASG, whose annotations imply venom-related functions. Despite the subsets of transcripts highly-expressed in the VG, SG, and ASG being very distinct, SG expresses an L-, and ASG-Cerm08-, and MEFRR- superfamily conotoxins, all previously considered specific for VG. We corroborate our results with the analysis of published SG and VG transcriptomes from unrelated fish-hunting C. geographus, and C. striatus, possibly fish-hunting C. rolani, and worm-hunting Conus quercinus. In spite of low expression levels of conotoxins, some other specific clusters of putative venom-related peptides are present and may be highly expressed in the SG of these species. Further functional studies are necessary to determine the role that these peptides play in envenomation. In the meantime, our results show importance of routine multi-tissue sampling both for accurate interpretation of tissue-specific venom composition in cone-snails, and for better understanding origin and evolution of venom peptides genes.

Systematics and Distributions of Upper Bathyal Species in Bathyancistrolepis, a Deep-Sea Whelk Genus Endemic to the Northwest Pacific (Gastropoda: Buccinidae).

The deep-sea buccinid snail genus Bathyancistrolepis is redefined based on the reconstruction of a molecular phylogeny and morphological examination of shell and radular characters. This genus is distinguished from other genera of the subfamily Parancistrolepidinae with a combination of shell traits, including (1) a low spire, (2) sharp, carinate spiral cords or keels and (3) a long, curved siphonal canal, but not with a difference in radular morphology as suggested by previous authors. Three allopatric or parapatric species are recognized in the upper bathyal (447-2057 m) waters around Japan and Taiwan: B. tokoyodaensis from off Hokkaido to Sagami Bay in the Northwest Pacific, B. trochoidea off Kumano-nada to Miyazaki in the Northwest Pacific and along Nansei Islands in the East China Sea, and B. taiwanensis sp. nov. in the South China Sea. These species bear large paucispiral protoconchs that are indicative of benthic early development without a pelagic larval period, and hence low dispersal capability. Seafloor topography seems to have acted as a barrier for their dispersal; the range of B. tokoyodaensis supports the previous finding that Izu Peninsula delimits westward distribution of bathyal gastropod species of boreal origins.

Exon-Capture-Based Phylogeny and Diversification of the Venomous Gastropods (Neogastropoda, Conoidea).

Transcriptome-based exon capture methods provide an approach to recover several hundred markers from genomic DNA, allowing for robust phylogenetic estimation at deep timescales. We applied this method to a highly diverse group of venomous marine snails, Conoidea, for which published phylogenetic trees remain mostly unresolved for the deeper nodes. We targeted 850 protein coding genes (678,322 bp) in ca. 120 samples, spanning all (except one) known families of Conoidea and a broad selection of non-Conoidea neogastropods. The capture was successful for most samples, although capture efficiency decreased when DNA libraries were of insufficient quality and/or quantity (dried samples or low starting DNA concentration) and when targeting the most divergent lineages. An average of 75.4% of proteins was recovered, and the resulting tree, reconstructed using both supermatrix (IQ-tree) and supertree (Astral-II, combined with the Weighted Statistical Binning method) approaches, are almost fully supported. A reconstructed fossil-calibrated tree dates the origin of Conoidea to the Lower Cretaceous. We provide descriptions for two new families. The phylogeny revealed in this study provides a robust framework to reinterpret changes in Conoidea anatomy through time. Finally, we used the phylogeny to test the impact of the venom gland and radular type on diversification rates. Our analyses revealed that repeated losses of the venom gland had no effect on diversification rates, while families with a breadth of radula types showed increases in diversification rates, thus suggesting that trophic ecology may have an impact on the evolution of Conoidea.

Delimiting species of marine gastropods (Turridae, Conoidea) using RAD sequencing in an integrative taxonomy framework.

Species delimitation in poorly known and diverse taxa is usually performed based on monolocus, DNA-barcoding-like approaches, while multilocus data are often used to test alternative species hypotheses in well-studied groups. We combined both approaches to delimit species in the Xenuroturris/Iotyrris complex, a group of venomous marine gastropods from the Indo-Pacific. First, COI sequences were analysed using three methods of species delimitation to propose primary species hypotheses. Second, RAD sequencing data were also obtained and a maximum-likelihood phylogenetic tree produced. We tested the impact of the level of missing data on the robustness of the phylogenetic tree obtained with the RAD-seq data. Alternative species partitions revealed with the COI data set were also tested using the RAD-seq data and the Bayes factor species delimitation method. The congruence between the species hypotheses proposed with the mitochondrial nuclear data sets, together with the morphological variability of the shell and the radula and the distribution pattern, was used to turn the primary species hypotheses into secondary species hypotheses. Allopatric primary species hypotheses defined with the COI gene were interpreted to correspond to intraspecific structure. Most of the species are found sympatrically in the Philippines, and only one is confidently identified as a new species and described as Iotyrris conotaxis n. sp. The results obtained demonstrate the efficiency of the combined monolocus/multilocus approach to delimit species.

A multilocus molecular phylogeny of Fasciolariidae (Neogastropoda: Buccinoidea).

The neogastropod family Fasciolariidae Gray, 1853 - tulips, horse-conchs, spindles, etc., comprises important representatives of tropical and subtropical molluscan assemblages, with over 500 species in the subfamilies Fasciolariinae Gray, 1853, Fusininae Wrigley, 1927 and Peristerniinae Tryon, 1880. Fasciolariids have had a rather complicated taxonomical history, with several genus names for a long time used as waste baskets to group many unrelated species; based on shell characters, recent taxonomic revisions have, however, began to set some order in its taxonomy. The present work is the first molecular approach to the phylogeny of Fasciolariidae based on a multigene dataset, which provides support for fasciolariids, an old group with a fossil record dating back to the Cretaceous. Molecular markers used were the mitochondrial genes 16S rRNA and cytochrome c oxidase subunit I, and the nuclear genes 18S rRNA, 28S rRNA and histone H3, sequenced for up to 116 ingroup taxa and 17 outgroups. Phylogenetic analyses revealed monophyly of Dolicholatirus Bellardi, 1884 and Teralatirus Coomans, 1965, however it was not possible to discern if the group is the sister clade to the remaining fasciolariids; the latter, on the other hand, proved monophyletic and contained highly supported groups. A first split grouped fusinines and Pseudolatirus Bellardi, 1884; a second split grouped the peristerniine genera Peristernia Mörch, 1852 and Fusolatirus Kuroda and Habe, 1971, while the last group comprised fasciolariines and the remaining peristerniines. None of these clades correspond to the present-day accepted circumscription of the three recognized subfamilies.

Lost and found: the Eocene family Pyramimitridae (Neogastropoda) discovered in the recent fauna of the Indo-Pacific.

Most neogastropod families have a continuous record from the Cretaceous or Paleogene to the Recent. However, the fossil record also contains a number of obscure nominal families with unusual shell characters that are not adequately placed in the current classification. Some of these are traditionally regarded as valid, and some have been "lost" in synonymy. One such "lost" family is the Pyramimitridae, established by Cossmann in 1901 for the Eocene genus Pyramimitra, and currently included in the synonymy of Buccinidae. Examination of several species of inconspicuous, small turriform gastropods has revealed a radula type so far unknown in Neogastropoda, and their shell characters identify them as members of the "extinct" family Pyramimitridae. Neither the radular morphology nor the anatomy reveal the relationships of this enigmatic, "living fossil" family. Molecular data (12S, 16S, 28S, COI) confirm the recognition of Pyramimitridae as a distinct family, but no sister group was identified in the analysis. The family Pyramimitridae Cossmann, 1901, is thus restored as a valid family of Neogastropoda that includes the genera Pyramimitra Conrad, 1865, Endiatoma Cossmann, 1896, Vaughanites Woodring, 1928, Hortia Lozouet, 1999, and Teremitra new genus. Pyramimitrids occur in the Recent fauna at bathyal depths of the Indo-Pacific from Taiwan to Madagascar and New Zealand, with three genera and nine species (all but one new).

Main patterns of radula formation and ontogeny in Gastropoda.

Gastropoda is morphologically highly variable and broadly distributed group of mollusks. Due to the high morphological and functional diversity of the feeding apparatus gastropods follow a broad range of feeding strategies: from detritivory to highly specialized predation. The feeding apparatus includes the buccal armaments: jaw(s) and radula. The radula comprises a chitinous ribbon with teeth arranged in transverse and longitudinal rows. A unique characteristic of the radula is its continuous renewal during the entire life of a mollusk. The teeth and the membrane are continuously synthesized in the blind end of the radular sac and are shifted forward to the working zone, while the teeth harden and are mineralized on the way. Despite the similarity of the general mechanism of the radula formation in gastropods, some phylogenetically determined features can be identified in different phylogenetic lineages. These mainly concern shape, size, and number of the odontoblasts forming a single tooth. The radular morphology depends on the shape of the formation zone and the morphology of the subradular epithelium. The radula first appears at the pre- and posttorsional veliger stages as an invagination of the buccal epithelium of the larval anterior gut. The larval radular sac is lined with uniform undifferentiated cells. Each major phylogenetic lineage is characterized by a specific larval radula type. Thus, the docoglossan radula of Patellogastropoda is characterized by initially three and then five teeth in a transverse row. The larval rhipidoglossan radula has seven teeth in a row with differentiation into central, lateral, and marginal teeth and later is transformed into the adult radula morphology by the addition of lateral and especially marginal teeth. The taenioglossan radula of Caenogastropoda is nearly immediately formed in adult configuration with seven teeth in a row.

The rhipidoglossan radula: Radular morphology and formation in Nerita litterata Gmelin, 1791 (Neritimorpha, Neritidae).

The rhipidoglossan radula, consisting of numerous teeth in each transverse row, is characteristic of phylogenetically distant groups of gastropods, including Vetigastropoda, Neritimorpha and 'lower' Heterobranchia. Previous studies have revealed the main patterns in the formation of the rhipidoglossan radula of vetigastropods, the main feature of which is the division of the formation zone into two horns, where marginal teeth are formed by a multilayered epithelium (odontoblasts). This work is devoted to the study of the formation of the rhipidoglossan radula of Nerita litterata using light and electron microscopy. The data obtained show that, despite the different external morphology of the radular sac of neritids and vetigastropods, the radular sac of N. litterata, like that of vetigastropods, is divided into two parts, in which the marginal teeth are similarly formed by odontoblasts located in more than one layer. It seems probable that this complex, three-dimensional structure of the formation zone is associated with a broad radula with numerous elongate marginal teeth and could be characteristic of other gastropods with this type of radula. Additional supporting rods located along the odontoblasts and consisting of vacuolated cells were first discovered in Nerita.

Neogastropod (Mollusca, Gastropoda) phylogeny: A step forward with mitogenomes.

The Neogastropoda (Mollusca, Gastropoda) encompass more than 15,000 described species of marine predators, including several model organisms in toxinology, embryology and physiology. However, their phylogenetic relationships remain mostly unresolved and their classification unstable. We took advantage of the many mitogenomes published in GenBank to produce a new molecular phylogeny of the neogastropods. We completed the taxon sampling by using an in-house bioinformatic pipeline to retrieve mitochondrial genes from 13 transcriptomes, corresponding to five families not represented in GenBank, for a final dataset of 113 taxa. Because mitogenomic data are prone to reconstruction artefacts, eight different evolutionary models were applied to reconstruct phylogenetic trees with IQTREE, RAxML and MrBayes. If the over-parametrization of some models produced trees with aberrant internal long branches, the global topology of the trees remained stable over models and softwares, and several relationships were revealed or found supported here for the first time. However, even if our dataset encompasses 60% of the valid families of neogastropods, some key taxa are missing and should be added in the future before proposing a revision of the classification of the neogastropods. Our study also demonstrates that even complex models struggle to satisfactorily handle the evolutionary history of mitogenomes, still leading to long-branch attractions in phylogenetic trees. Other approaches, such as reduced-genome strategies, must be envisaged to fully resolve the neogastropod phylogeny.

The rhipidoglossan radula: Formation and development in Margarites helicinus Phipps, 1774 (Trochoidea, Vetigastropoda).

The gastropod radula exhibits exceptional morphological variability. Despite this enormous diversity, the main patterns of synthesis of the teeth and radula membrane, characteristic of different groups can be identified. The rhipidoglossan radula contains numerous teeth in each transverse row and was found in phylogenetically distant groups of Gastropoda. Studying radula formation through stages of ontogeny in species with this type of radula is important in determining the main patterns of radula formation in gastropods. We studied the formation of the radula during development of one species of trochid vetigastropod, Margarites helicinus, using light and electron microscopy as well as confocal laser scanning microscopy. The adult radula is formed in the blind end of the radular sac, which bifurcates into two horns in vetigastropods. The numerous marginal teeth are synthesized in these horns while the central and lateral teeth form in the region where the horns fuse. This morphology of the formation zone appears to be a common pattern for all vetigastropods. The juvenile radula of M. helicinus consists of seven teeth per transverse row and its formation in the radular sac differs significantly from that in the adult. In the juvenile, the formation zone of the radular sac is not split into two horns, and the teeth and radular membrane are synthesized by relatively few, uniform cells. This organization of the larval radular sac is thought to represent a widely occurring larval pattern potentially present in all groups of gastropods. It is associated with early formation of all organs by few cells rather than representing a phylogenetic trait.

The rhipidoglossan radula: Formation and morphology of the radula in Puncturella noachina (Linnaeus, 1771) (Fissurellidae, Vetigastropoda).

The rhipidoglossan radula, which is characterized by presence of a central tooth, several lateral teeth, and numerous (more than 10) long marginal teeth in each transverse row, is found in three different subclasses, that is, Vetigastropoda, Neritomorpha and "lower" Heterobranchia. Details of radula formation and its ultrastructure have not been studied in any species with a rhipidoglossan radula. For the first time, we present such data for one vetigastropod species, Puncturella noachina. The radula itself and the radula formation zone were studied using light and electron microscopy (scanning and transmission), as well as confocal laser scanning microscopy. We identify the major features of Vetigastropoda rhipidoglossan radula formation, that is: the posterior bifurcation of the radula formation zone, creating paired horns into which the zones of formation of the marginal teeth extend; the supporting structure in the radula formation zone extends ventrally to strengthen this division; the odontoblasts of the marginal teeth form a multi-layered epithelium; membranoblasts do not differ from odontoblasts in ultrastructure; in some membranoblasts and cells of the sub- and supraradular epithelium basal bodies were found in the apical regions of the cells.

Radula formation in two species of Conoidea (Gastropoda).

The radula is the basic feeding structure in gastropod molluscs and exhibits great morphological diversity that reflects the exceptional anatomical and ecological diversity occurring in these animals. This uniquely molluscan structure is formed in the blind end of the radular sac by specialized cells (membranoblasts and odontoblasts). Secretion type, and the number and shape of the odontoblasts that form each tooth characterize the mode of radula formation. These characteristics vary in different groups of gastropods. Elucidation of this diversity is key to identifying the main patterns of radula formation in Gastropoda. Of particular interest would be a phylogenetically closely related group that is characterized by high variability of the radula. One such group is the large monophyletic superfamily Conoidea, the radula of which is highly variable and may consist of the radular membrane with five teeth per row, or the radular membrane with only two or three teeth per row, or even just two harpoon-like teeth per row without a radular membrane. We studied the radulae of two species of Conoidea (Clavus maestratii Kilburn, Fedosov & Kantor, 2014 [Drilliidae] and, Lophiotoma acuta (Perry, 1811) [Turridae]) using light and electron microscopy. Based on these data and previous studies, we identify the general patterns of the radula formation for all Conoidea: the dorsolateral position of two groups of odontoblasts, uniform size, and shape of odontoblasts, folding of the radula in the radular sac regardless of the radula configuration. The morphology of the subradular epithelium is most likely adaptive to the radula type.

Venom Diversity and Evolution in the Most Divergent Cone Snail Genus Profundiconus.

Profundiconus is the most divergent cone snail genus and its unique phylogenetic position, sister to the rest of the family Conidae, makes it a key taxon for examining venom evolution and diversity. Venom gland and foot transcriptomes of Profundiconus cf. vaubani and Profundiconusneocaledonicus were de novo assembled, annotated, and analyzed for differential expression. One hundred and thirty-seven venom components were identified from P. cf. vaubani and 82 from P. neocaledonicus, with only four shared by both species. The majority of the transcript diversity was composed of putative peptides, including conotoxins, profunditoxins, turripeptides, insulin, and prohormone-4. However, there were also a significant percentage of other putative venom components such as chymotrypsin and L-rhamnose-binding lectin. The large majority of conotoxins appeared to be from new gene superfamilies, three of which are highly different from previously reported venom peptide toxins. Their low conotoxin diversity and the type of insulin found suggested that these species, for which no ecological information are available, have a worm or molluscan diet associated with a narrow dietary breadth. Our results indicate that Profundiconus venom is highly distinct from that of other cone snails, and therefore important for examining venom evolution in the Conidae family.

Alpha-conopeptides specifically expressed in the salivary gland of Conus pulicarius.

To date, studies conducted on cone snail venoms have attributed the origins of this complex mixture of neuroactive peptides entirely to gene expression by the secretory cells lining the lumen of the venom duct. However, specialized tissues such as the salivary glands also secrete their contents into the anterior gut and could potentially contribute some venom components injected into target animals; evidence supporting this possibility is reported here. Sequence analysis of a cDNA library created from a salivary gland of Conus pulicarius revealed the expression of two transcripts whose predicted gene products, after post-translational processing, strikingly resemble mature conopeptides belonging to the alpha-conotoxin family. These two transcripts, like alpha-conotoxin transcripts, putatively encode mature peptides containing the conserved A-superfamily cysteine pattern (CC-C-C) but the highly conserved A-superfamily signal sequences were not present. Analysis of A-superfamily members expressed in the venom duct of the same C. pulicarius specimens revealed three putative alpha-conotoxin sequences; the salivary gland transcripts were not found in the venom duct cDNA library, suggesting that these alpha-conotoxins are salivary gland specific. Therefore, expression of conotoxin-like gene products by the salivary gland could potentially add to the complexity of Conus venoms.

Morphological proxies for taxonomic decision in turrids (mollusca, neogastropoda): a test of the value of shell and radula characters using molecular data.

The state of the art of turrid (=Turridae s. l.) systematics is that shells - when they include the protoconch - are reliable species-level identifiers, but inadequate proxies for allocation to genera or subfamilies. Generally, the radula is used for allocation to a (sub)family, but the hypothesis that the radula is a more adequate proxy than the shell for relationships has not yet been tested by molecular data. Species of Xenuroturris may have drastically different radulae, with either "semi-enrolled" or "duplex" marginal teeth, although their shells are very similar or even almost indistinguishable. Molecular data confirm that specimens with different types of radulae constitute different species, but two species of a pair with respectively semi-enrolled and duplex teeth end up being not closely related. However, it is still unresolved whether species with semi-enrolled (= Iotyrris ) and duplex teeth (= Xenuroturris ) form two supported monophyletic clades. Iotyrris devoizei n.sp. and I. musivum n.sp. are described from Vanuatu, where they occur sympatrically with I. cingulifera and Xenuroturris legitima .

Quid est Clea helena? Evidence for a previously unrecognized radiation of assassin snails (Gastropoda: Buccinoidea: Nassariidae).

The genus Clea from SE Asia is from one of only two unrelated families among the megadiverse predatory marine Neogastropoda to have successfully conquered continental waters. While little is known about their anatomy, life history and ecology, interest has grown exponentially in recent years owing to their increasing popularity as aquarium pets. However, the systematic affinities of the genus and the validity of the included species have not been robustly explored. Differences in shell, operculum and radula characters support separation of Clea as presently defined into two distinct genera: Clea, for the type species Clea nigricans and its allies, and Anentome for Clea helena and allies. A five-gene mitochondrial (COI, 16S, 12S) and nuclear (H3, 28S) gene dataset confirms the placement of Anentome as a somewhat isolated offshoot of the family Nassariidae and sister to the estuarine Nassodonta. Anatomical data corroborate this grouping and, in conjunction with their phylogenetic placement, support their recognition as a new subfamily, the Anentominae. The assassin snail Anentome helena, a popular import through the aquarium trade so named for their voracious appetite for other snails, is found to comprise a complex of at least four species. None of these likely represents true Anentome helena described from Java, including a specimen purchased through the aquarium trade under this name in the US and one that was recently found introduced in Singapore, both of which were supported as conspecific with a species from Thailand. The introduction of Anentome "helena" through the aquarium trade constitutes a significant threat to native aquatic snail faunas which are often already highly imperiled. Comprehensive systematic revision of this previously unrecognized species complex is urgently needed to facilitate communication and manage this emerging threat.

Cheсklist of gastropod molluscs in mangroves of Khanh Hoa province, Vietnam.

Gastropod molluscs are one of the most important components of mangrove ecosystem. Mangroves in Central Vietnam have a rather limited distribution due to peculiarities of the coastline morphology and presently their fauna remains understudied. Extensive surveys were conducted in both natural vegetation and artificial mangrove plantations in several localities in Nha Trang Bay from 2005 to 2015. In total 65 species of gastropod molluscs were found alive, 17 of which can be considered as predominantly mangrove-associated. An illustrated guide is provided, with short synonymies and data on ecology and distribution. The recorded molluscan diversity is compared with published data on mangrove gastropods in different regions of the Indo-Pacific. Total species number and the proportion of mangrove-associated species are similar to studied faunas in Hong Kong, Malaysia and Thailand, but the diversity is much lower than that of the mangal fauna of the Philippines.