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This approach was used to identify 70 bp is needed for efficient expression of most genes, potential introns using similarity searches with the EST even if they are stage-specific [38]. The S. Given that the transcriptional start site and are essential and suffi- only a few introns have been detected within the diplo- cient for promoter activity [39]. However, eukaryotic promoter motifs Figure 2A. However, AT-rich two introns were detected in one of the first genes stretches are found close to the translational start sites, or sequenced from Carpediemonas [44], an excavate taxon in the first 50 bp upstream Figure 2A and the intergenic and possible sister lineage to diplomonads.

Thus, introns regions are very short Figure 1. Thus, the S. A basic spliceosomal organisation is present in Gia- tion codon could be mapped in EST clones. The 3′ rdia suggesting that the spliceosome is ancestral to extant untranslated regions UTR of S. Although our analyses did not identify to be short; the average distance between the termination any proteins likely to be involved in splicing in S.

Such very short 3′ UTRs have pre- spliceosomal apparatus are present in S. In genes, distributed among all functional categories, repre- G. A: 5′ regions of full-length genes from the completely sequenced contigs. The initiation codons of nine genes in the contigs Figure 1 could be precisely identified based on sequence conservation com- pared to orthologs.

The 5′ sequences are aligned based on the initiation codons indicated by a box. Among these were genes involved in transla- tRNA. The identification of a putative Sec tRNA, which is tion: 74 genes encoding ribosomal proteins, many trans- a central component of selenoprotein biosynthesis [47], lation factors, and fifteen different tRNA synthetases were in our data set suggests that Spironucleus is able to use the identified Additional file 2.

Twenty tRNA genes were single stop codon UGA to incorporate this rare amino identified, which cover 24 of the 63 sense codons allow- acid into selenoproteins. Indeed, the usage of seleno- ing for the normal wobble rules for codon-anticodon cysteine seems to be widespread feature in protists; Sec pairing.

Interestingly, all 64 codons appear to have the The identified proteins within the cellular, environmental potential to code for incorporation of amino acids into information, and genetic information process categories proteins in S.

This is [28] and a cysteine desulfurase called Nifsp in Saccharo- indeed expected from the current view of diplomonad myces cerevisiae , a key enzyme of the Fe-S cluster synthesis phylogeny and cell biology see Background section. Orthologous proteins of Interestingly, only eight Rab genes were found in G.

Localization studies of these two proteins should more-complex endomembrane system in this parabasalid indeed be very useful to investigate whether S. Two of the five Rab proteins identified in our survey contains mitosomes.

The G. The function of the G. Yet, a For example, relatively few enzymes involved in amino single dynamin-like protein encoding gene is also found acid metabolism were detected Additional file 2.

Ami- in three kinetoplastid genomes two Trypanosoma and one noacyl-tRNA synthetases are classified into this category, Leishmania [55] and the microsporidium Encephalitozoon but they are atypical since they are essential for protein cuniculi [56], whereas most eukaryotic genomes encode synthesis. Furthermore, only a single protein malate several dynamin-like paralogues that function either in dehydrogenase associated with the tricarboxylic acid membrane trafficking or organelle division mitochon- TCA cycle was found.

However, malate dehydrogenase dria and plastids [57]. Interestingly, the single dynamin- actually functions in a pyruvate synthesis pathway in G. Hence the single ilar role in S. In contrast, several glycolytic dynamin-like proteins in Spironucleus, Giardia and proteins are present in our data set Additional file 2.

Encephalitozoon could all be involved in mitosome divi- This pattern of metabolic proteins is expected from a fer- sion.

The predicted amino acid sequences of genes were These similarities between S. Most of these were anno- such an environment in the two lineages. Indeed, a con- tated as conserved hypothetical proteins, although some siderable fraction of the metabolic proteins was found to could be assigned to a functional category Additional file be more closely related to prokaryotic rather than eukary- 2. Most of the cysteine residues were found as CXXC otic homologs in the phylogenomic analyses, suggesting motifs Figure 3 and data not shown.

Such an arrange- acquisition of these genes by gene transfer see further dis- ment is similar to the large protein family of variant-spe- cussion below. Does S. A new organelle, the mitosome, has recently been identi- salmonicida showed highest sequence similarities to G. Currently, the only known function of motifs of G. The phylogenetic relationship between G. Indeed, in similarity and S. These observa- present in all extent eukaryotes [10] strongly suggest that tions indicate that diplomonads vary greatly in their the common ancestor of the two species contained an cysteine-rich proteins; the gene families indeed appear to organelle with mitochondrial ancestry.

Among the cysteine-rich proteins coding sequences two are likely to represent full-length ORFs. These are In the laboratory, Spironucleus muris is transmitted as cysts strong candidate surface proteins since they possess a in fecal material like Giardia [66]. Cysts of S. A total of 14 even display the same immunostaining as Giardia cysts sequences encoding partial proteins with cysteine-rich when labeled with antibodies specific for Giardia cyst wall domains possess similar structural organization as the [66].

We identified one homolog of G. No TM domains could be found in the remaining phosphate isomerase and UDP-N-acetylglucosamine cysteine-rich proteins within our dataset. However, in sev- pyrophosphorylase — Additional file 2. The presence of eral cases these partial cysteine-rich proteins show high these three genes indicate that S. This initiates infection in fish.

These genes are stage-specifically observation is consistent with the genome of S. Characteristic features in G. None of the at a relatively high frequency, which is important for genes encoding disc-specific proteins in G. In contrast to G. Cysteine proteases are essential for pathogenicity pathogenicity and invasion of the intestine of E. There- Spironucleus species have been described as a cause of dis- fore the several cysteine proteases that were found in our ease in hosts that include birds, fish and mice [1]; S.

However, essentially nothing is known about viru- Secretion of cysteine proteases was indeed recently shown lence factors in Spironucleus. In healthy fish, the parasite to occur from trophozoites of G. Cysteine protease activities are stage trophozoites in the lumen of the upper intestine, also important for excystation and encystation of Giardia where it remains attached to the intestinal mucosa, con- [70,71], suggesting yet another potential role for these trolled by the mucosal immune system of the host.

This enzymes in S. Antigenic variation among the identified Genome-wide mutational processes have been identified cysteine-rich proteins in S. Nitric oxide NO and reactive oxygen tions within genomes may be explained by selection species ROS are important factors in the host’s protec- [72,73]. We analyzed sequences of genes to examine tion against mucosal pathogens [62]. The random sequencing of ESTs from a non- teins and arginine deiminase Additional file 2.

These normalized cDNA library provided a rough estimate of proteins, which are coded by genes putatively acquired the expression levels of S. The dia- gram shows the position of the seven furin-like domains SM — orange boxes relative the transmembrane domain blue box as determined by SMART4.

Due to the overlap between these infer- ences only the furin-like domains are shown for simplicity. This is consistent with the hypothesis that cysteine-rich domains are facing the extracellular milieu where they could interact with other proteins.

The alignment of the seven inferred furin-like domains as inferred by T-COFFEE [] is shown below the diagram, with the positions of the first and last residue of each domain indicated on the left.

Cysteines are highlighted in red and bold. No putative signal peptide was found with SignalP3. This protein has a similar structural organization as the one shown in A with three EGF-like domains inferred to face the external milieu. The three EGF-like domains were aligned manually. In addition, this sequence may have a signal peptide indicated by a red box since the S-score is positive in SignalP3.

C: Domain organization of G. All dia- grams are drawn to scale. A mates of expression levels were available for a few of the distinct pattern was found for genes never found in the genes. Our study provides, for a much larger dataset, a EST library “weakly expressed” , for which codon usage connection between the subset of genes with a strong is more uniform, with no codons strongly dominating.

To codon bias, and an indirect indication of expression levels compare our results with a previous study from Giardia cDNA abundance , corroborating the earlier conclusion [74], G. In salmonicida. The analysis Interestingly, the favored codons differed between S.

In nine of highly expressed genes, while the putatively weakly the cases where there is a difference between the two spe- expressed genes have a more uniform amino acid usage. The tenth case is a two-fold salmonicida genes degenerate amino acid where S. There is a mous position GC3s , and performed correspondence distinction between the base compositional biases in G. This is an expected pat- Figure 4A.

For G. However, a differences seem to have influenced the codon preferences separation of the genes into different categories can be in the two diplomonad lineages in the highly expressed observed: the vast majority of the weakly expressed genes genes as well. In the absence of information about codon have GC3s values between 0. At any rate, the usage of an the third synonymous position is the main determinant of alternative genetic code in Spironucleus but not Giardia the codon usage in both S.

Expres- glutamine codons. The canonical stop codon UAA is sion levels strongly influence the second axis in the corre- indeed the most common codon for glutamine in our spondence analysis for S. No such trend is observed for G. These weakly expressed genes in S. Unexpectedly, analyses clearly indicate that there is selection on codon the Nc’ values for genes with GC3s above 0.

Translation genes, the Nc’ values should be close to the maximum. Genes are represented by different colors and symbols according to the number of times they were sampled within the EST data.

Genes identified to be involved in gene transfer and are shared with G. If the observed codon usage pattern is more uniform than expected by chance, the Nc’ value is set to 61 [75, ].

The candidate LGTs unique to S. Also, the large spread of the genes to the Although some genes with high GC3s values do show left on axis 1 which is strongly correlated with GC3s — indications of gene transfer in the phylogenomic analyses, Figure 4C argues against selection for an alternative set the majority do not Figure 4A, C. In fact, many of these of optimal codons in these genes. Cases where two S. Thus, Additional Files 3, 4, 5, 6. Furthermore, on average five genes with unexpected codon usage i.

How- tion codons in all characterized prokaryotes [84] were ever, neither mutational or selection scenarios can easily detected in 21 of the remaining 27 putatively transferred explain their non-random codon usage, nor the fluctua- genes, strongly suggesting a true S.

Heterogeneity in for these genes. For S. We cannot formally siae a unicellular eukaryote with a compact genome, that exclude contamination on the basis of presence of alterna- is more directly comparable to S. At any rate, together these observations rience different chemical environments [see [78] and ref- indicate that the phylogenetic pattern we interpret as erences therein]. Longer genomic fragments, as well as putative LGT events are unlikely to result from substantial comparative data from more-closely related diplomon- contamination of our libraries.

Some Spironucleus genes were strongly associated Phylogenomic analysis reveals frequent lateral gene with genes from distantly related organisms. For example, transfer the S.

LGT in diplomonads on a single-gene basis [16,17,], Such anomalous positions of the diplomonads genes []. Here we present a systematic phylogenomic anal- strongly suggest gene transfer events.

Many datasets con- ysis of the complete EST and GSS data sets with the goal tain sequences from only one or a few eukaryotic taxa of identifying S. We used the PhyloGenie package [83] to assemble diplomonads are the only eukaryotes present Figure 5B— aligned amino acid data sets automatically from databases D.

Although the separation of diplomonad from the including both published sequences and data from ongo- other eukaryotic sequences is sometimes weak i. Figure ing eukaryotic genome projects see Methods. PCC Clostridium tetani Nostoc punctiforme Clostridium thermocellum 85 Synechococcus elongatus 64 84 Trichomonas vaginalis 84 Trichomonas vaginalis 73 Synechococcus sp.

WH 97 99 Prochlorococcus marinus subsp. Delta H 54 uncultured proteobacterium QS1 Methanococcus maripaludis Entamoeba histolytica 61 Nocardia farcinica unknown taxonomy Chromobacterium violaceum Wolinella succinogenes 61 Nostoc punctiforme Thermoanaerobacter tengcongensis 92 Trichodesmium erythraeum Archaeoglobus fulgidus 68 Thermococcus kodakaraensis Crocosphaera watsonii 70 Thermotoga maritima 60 Synechocystis sp.

PCC Clostridium acetobutylicum 96 Thermosynechococcus elongatus 95 Clostridium tetani Gloeobacter violaceus Clostridium thermocellum 62 64 Methanosarcina barkeri Prochlorococcus marinus Methanosarcina acetivorans Homo sapiens Archaeoglobus fulgidus 95 Macaca fascicularis Chlorobium tepidum 88 Tetraodon nigroviridis Magnetococcus sp. MC-1 Dehalococcoides ethenogenes Tetraodon nigroviridis Geobacter metallireducens Tetraodon nigroviridis 59 Methanothermobacter thermautotrophicus 86 Thalassiosira pseudonana 75 Methanosarcina acetivorans Tetrahymena thermophila 90 Methanococcoides burtonii 78 Desulfovibrio vulgaris 94 Thalassiosira pseudonana Geobacter sulfurreducens Anabaena variabilis Desulfitobacterium hafniense 60 Gloeobacter violaceus Desulfovibrio desulfuricans uncultured proteobacterium QS1 53 95 Synechococcus sp.

ML tree of conceptually translated, aligned amino acid positions of A arginine deiminase, B conserved hypothetical protein, C conserved hypothetical protein, D carotenoid isomerase, and E rubrerythrin. Details about the phylogenetic analyses are found in the Methods section, and complete accession numbers and complete species names are found in Addi- tional files 4 and 6.

The unrooted trees are arbitrarily rooted for the presentation. Eukaryotes are in boldface and labeled according to their classification into super-groups [18]: Opisthokonta orange , Amoebozoa purple , Chromalveolata red , Archaeplastida green , and Excavata brown. The genes for two enzymes in the pathway, arginine deiminase and ornithine transcar- There are indeed examples of inferences of LGT based on bamoylase, appear to have been acquired from prokaryo- unexpected patterns of phyletic distribution that have pre- tes in a common ancestor of diplomonads and viously been shown to be wrong.

Re-analyses of the puta- parabasalids, whereas carbamate kinase may have distinct tive bacteria-to-vertebrate gene transfer events in the LGT origins in the two groups Figure 5A and trees 10 human genome using phylogenetic analyses instead of and 30 in Additional file 4.

Similarly, S. Phylogenetic analyses may also defense see above [62,64,65] that are also found in lead to false positive interpretations of LGT. For example, anaerobic prokaryotes and eukaryotes including Enta- phylogenetic analysis indicated an LGT origin of the T. On the other hand, re- advantages to the recipient protist lineages.

Indeed, dis- analyses using extended taxon sampling and more tantly related lineages that live in the same environment advanced models of sequence evolution of four genes appear to frequently transfer genes [89].

Obviously, additional acquired metabolic genes Additional file 3. Therefore, it is inevitable that the list eral origin, and in previous phylogenomic studies 96 E. Nevertheless, our Cryptosporidium parvum and genes in anaerobic cili- phylogenomic analysis indicates that vertical inheritance ates were found to represent candidate LGT genes combined with gene duplication and gene loss is rather [51,55,91,92].

Thus, numerous inter-domain gene trans- unlikely to have produced the observed phylogenetic rela- fer events have been identified in four divergent protist tionships and patchy taxonomic distributions and that lineages — Spironucleus, Entamoeba, kinetoplastids gene transfer is the simplest hypothesis currently available Trypanosoma and Leishmania and anaerobic ciliates — to explain these trees.

Hence our analyses indicate that supporting the idea that gene acquisition from prokaryo- LGT has been an important mechanism in the evolution tic organisms is a common evolutionary mechanism in of the S.

In all these studies, metabolic genes have been found to be the most common Acquisitions of prokaryotic metabolic genes in the functional category among the genes implicated in LGT evolution of protist genomes genes.

This bias is not unexpected, as a higher rate of transfer for metabolic genes compared to informational genes has G. In the events, while no closely related G. For example, amino rdia lineages. The remaining 19 events could have taken acids have been identified to be a source of energy in place in the S. Among losses in the latter could also explain the observed pattern. Additional eukaryote-to-eukaryote other S.

Some in fact do show deviant transfers may actually have occurred among the genes that codon usage indices Figure 5A, B suggesting a more did not show any indication of prokaryote-to-eukaryote recent acquisition compared to the other cases.

However, transfer. A few examples of gene transfers between unicel- there are many other S. The 13 putative intra-domain ure 4A, B , suggesting that codon usage pattern alone is a transfers identified here further suggest that LGT between poor indicator of LGT in S. In 12 cases where both Spironucleus and Gia- neglected. This Conclusion pattern has previously been observed for candidate later- The vast majority of eukaryotic diversity is represented by ally transferred genes [16,17,80] and almost certainly protists [18], yet only a few protist genome sequencing reflects a common ancestry of diplomonads and paraba- projects have been published.

Our sequence survey study salids to the exclusion of other sampled eukaryotic line- has indicated that a combined approach using both ran- ages, although the possibility of gene transfer between the dom sampling from the genome GSSs and ESTs is suc- two lineages cannot be formally excluded. However, a cessful in identifying genes Table 1. We identified diplomonad-parabasalia relationship has recently been genes from the GSS sequences, while genes with shown to be robust in several phylogenetic analyses of homologies in other organisms were detected among the concatenated protein sequences [].

As some EST sequences. As we have collected more than twice as acquired genes are unique to S. However, EST LGT in these excavate lineages continuously throughout sequencing is biased towards identification of highly their evolutionary history. Further characterization of the expressed genes, such as genes involved in genetic infor- phylogenetic distribution of these genes may provide mation processing, especially translation, while GSS information about organismal relationships within Exca- sequencing detects a more random selection of genes vata [99].

Table 1. This functional bias of the genes detected in EST surveys may be an advantage if the objective of the study In the four previous phylogenomic studies of LGT in pro- is mainly to identify genes previously identified in other tists [51,55,91,92] only prokaryote-to-eukaryote transfer organisms using the same approach, but is a limitation if events were examined.

Our selection procedure allowed sampling gene diversity is the aim. In combination with ads and parabasalids, and then transferred to the lineage complete sequencing of a few contigs, this approach was leading to Dictyostelium Figure 5A.

Seventeen genes were efficient in revealing much about the S. Although we could identify more than genes to 13 unique events Additional file 3. Twelve of these with annotated functions, conserved hypothetical pro- cases are exchanges between the lineage leading to S. Furthermore, with the Entamoeba lineage, and a single case with the 13 among the 38 genes identified within the contigs did Mastigamoeba lineage Figure 5A, Additional files 3, 4, 5, not show any significant sequence similarity to genes in 6.

This suggests that a large fraction with genome sequences, and several diverse and com- of the genes in S. It is plete G. Sequencing identified individual S. A studied eukaryotes, including its closest intensively stud- genomic DNA library was constructed for GSS sequenc- ied relative, the diplomonad G. Mass in are diverse. For example, in the S. Cells were picked and the plasmids S. Roughly half of the sequencing was enteromonads and retortamonads, organisms with dis- done from this library.

However, since it was revealed that tinct morphological features [23,24,]. The DNA was physically G. One Shot of sequence divergence between members of the group, chemically competent cells Invitrogen were transformed and, as indicated here, large variation in the genomic with an aliquot of the ligation mix and spread on LB structure and content. The plasmids were quantified and sequenced genome sequencing project. Such an effort would yield as described above using T3 and T7 primers.

Messenger RNA from approximately macia. The ligations were transformed into XL2-Blue cells of S. Infection of SolR cells was carried out and Sequencing reactions were initially screened manually for plated on selective LB media containing carbenicillin, iso- exclusion of unsuccessful reactions. Positive colonies were respectively, passed this initial screen. The average read picked manually and plasmid DNA was purified using lengths of these were , and bp, respectively.

This procedure resulted in assembler [] with the default settings. The contigs were eight continuous stretches of genomic sequence between quality trimmed manually, while the singletons were and bp in length. Together with the three trimmed using the Phred software [] with the default sequenced lambda clones these yielded bp of fin- quality cut-off. Hybrid clones were identified as contain- ished genomic sequence in eleven contigs which were cov- ing stretches of nine or more As within the insert, and only ered by at least one-fold high quality sequence in each the 5′ end of the hybrid was retained.

The remain- in the wrong direction to known genes 12 ; these were ing sequences were treated as single reads, and reversed. Finally, nine EST sequences were trimmed covered bp of non-unique sequence after quality according to putative frame-shifts detected in similarity trimming.

After removal of contigs shorter than bases sequences contigs and singletons corre- To estimate the amount of unique high-quality sequence sponding to unique bp and based on clones within our survey project, we performed assemblies with remained for further analysis. The GSS clones yielded bp in contigs see below , respectively, and were excluded, leav- contigs with one or more sequences, and the addi- ing ESTs for further analyses. Lander and Waterman proposed that ESTs ended with a stretch of eight or more As, indica- the number of “islands” E in a genomic project with per- tive of mRNA polyadenylation, were identified.

All , the genome of S. The GSS chromatograms that passed this screen no host contamination was detected , and repre- Databases sented sequences longer than bp, were included in All databases used in the analyses were downloaded in the assembly.

The average read lengths of the clones from January The Phrap assembler was used with the from various nearly completed genome sequencing default settings, yielding contigs consisting of two or projects of diverse eukaryotes were downloaded: T.

Some of the longer nalis parabasalid , Tetrahymena thermophila ciliate , contigs were selected for complete sequencing. Regions Trypanosoma brucei, and Trypanosoma cruzi kinetoplastids that required additional sequencing were identified from the Institute for Genomic Research TIGR []; within these contigs, and amplified from genomic DNA Dictyostelium discoideum mycetozoan from dictyBase using PCR with primers designed based on the GSS assem- []; Chlamydomonas reinhardtii green alga , Phytoph- bled sequence.

To extend the contigs, sequence gaps covered by purposes. Thus, in total unique protein- nucleotide sequences from the single-read GSS and the coding genes were identified and included in further anal- EST contigs against the amino acid sequence databases yses.

Using the indication of the frame and direction bp or longer and were included in the codon-usage from the alignment between the query and database analysis.

Codon-usage indices were calculated for each sequence in the BLAST result files, the putative coding gene. Nc is a measure of the effective number of codons DNA and amino acid sequences were extracted from these used in a gene []. A modified version, Nc’, has been sequences. The single-read GSS sequences and developed which also account for the background nucle- EST contigs that failed to yield a match better than e-5 were otide composition of the gene; this is advantageous in sit- not analyzed further.

The Nc’ values were calculated using the To identify coding sequences in the finished contigs, sim- software INCA []. Met, Trp and the EST sequences were performed using the same and termination codons are excluded — were calculated cutoff as above. Similarity searches on the nucleotide level using the program CodonW []. The variation of codon against the non-redundant nucleotide databases were per- usage among genes was explored using the correspond- formed at NCBI to identify non-protein coding sequences.

To avoid identifica- Finally, open reading frames longer than bp with the tion of trends in codon usage due to biased amino acid expected GC pattern in the three coding positions were usage among the genes, the correspondence analysis was annotated as putatively coding and included in subse- performed on the relative synonymous codon usage quent analyses.

This procedure identified 38 protein-cod- RSCU values for each gene. The RSCU value for a codon ing genes in the eleven finished contigs. This should remove the effects of amino canSE version 1. For 19 of these, both codon, respectively.

The correspondence analysis plots programs assigned the same anticodon, while one dif- genes according to their RSCU values in a dimensional fered in anticodon identity see Results. Two tRNAs were space, and then identifies the major trends as the axes found to encode putative introns.

No tmRNA was found. To examine the variation of codon usage within diplo- monads, similarity searches of the S. These were considered homologs not necessarily genes that contained only the longest amino acid orthologs and the nucleotide sequences were obtained sequence from each gene among the GSS, EST, finished for codon-usage analysis as described above.

Three of the 30 previously published genes were represented by long ESTs The number of times each gene was detected in the EST and were excluded. This procedure resulted in unique library was used as an indication of expression level in the protein-coding genes. Codon-usage tables bp. Furthermore, of the remaining genes were calculated for each of these groups, both for S.

The matches with e values Gene annotation below e-5 were searched against the total yeast proteome The Kyoto Encyclopedia of Genes and Genomes KEGG with the same cutoff, which identified 71 reciprocal best project [], a bioinformatic knowledge base for system- matches between the datasets.

Based on the quality of the atic analysis of gene functions [], was used for gene BLASTp hit or protein length, to short to give any confi- annotation. One goal of this project was to identify genes in the S.

It takes amino acid sequences in standard fasta and an additional had non-best hits with assigned KO format, and performs similarity searches BLAST against numbers. Functional annotations were extracted from the a protein sequence database consisting of publicly availa- KO database for these genes. Similarity searches against all selected threshold are included in the dataset. Finally, an databases were performed for the remaining genes, alignment of these sequences is made using the profile.

No the datasets. In addition, the genes which had nomic analysis, since they were found to be problematic best hits with E values between e and e-5 were annotated to align probably due to their frequent possession of as conserved hypothetical proteins, and the remaining 13 highly repetitive sequences. PhyloGenie was run on the genes that did not have any match better than e-5 were annotated non-cysteine-rich genes with homologs annotated as hypothetical proteins.

The annotations for all proteins 1 since coverage is meaningless with partial gene mentioned in the text have been refined manually. Where more than amino acid sequences was found to vary considerably sequences pass the E-value cutoff, the with the lowest between genes. While the majority of genes would encode E-values in the HMM-based search will be retained. These were annotated as cysteine-rich pro- sequences from the dataset based on the taxonomic level teins. Bacillus sequences above a set identity.

DSH grew the These settings yielded datasets with four sequences or organism, purified nucleic acids, constructed the EST more, among which included or more aligned library, and carried out some of the initial EST sequencing. Only these were analyzed further, to avoid mis- carried out the majority of the EST sequencing, the initial leading results based on too-short alignments. A Perl GSS sequencing and the sequencing and assembly of three script provided in the PhyloGenie package [83] was mod- lambda clones.

SGS provided advice on the analyses of version 2. Production of gold and ruby-red lustres in Gubbio Umbria , Italy during the Renaissance period. The aim of this work is to gain a further insight into the knowledge of the production process of lustre-decorated ancient majolicas.

Lustre is a sophisticated technique employed in the decoration of majolicas as used in central Italy during the Renaissance period. It consists of a beautiful iridescent gold or ruby-red thin metallic film, containing silver, copper and other substances and obtained in a reducing atmosphere on a previously glazed ceramic. Nowadays, it is not possible to replicate the outstanding results obtained by the ancient ceramicists, since the original recipes were lost.

In this work, we have focussed our attention on ceramic fragments decorated with both gold and ruby-red lustres, which were difficult to obtain due to complex reduction conditions required and which were a prerogative of Gubbio production. The two lustre colours differ in their chemical composition as well in their nanostructure. The presence of bismuth was disclosed and it was ascertained to be a distinctive feature of the Italian production.

An integrated bio-chemostratigraphic framework for Lower Cretaceous Barremian-Cenomanian shallow-water carbonates of the Central Apennines Italy. Shallow-water carbonate platform sections are valuable archives for the reconstruction of deep-time environmental and climatic conditions, but the biostratigraphic resolution is often rather low. Moreover, chemostratigraphic correlation with well-dated pelagic sections by means of bulk carbonate carbon-isotope stratigraphy is notoriously difficult and afflicted with large uncertainties, as shallow-water sections are particularly prone to the impact of diagenesis.

In the current study, an integrated biostratigraphic-chemostratigraphic approach is applied to southern Tethyan Lower Cretaceous carbonate platform deposits Santa Lucia, Monte La Costa sections situated in the Central Apennines in Italy. The m thick Santa Lucia section, representing an open lagoonal inner carbonate platform setting, provides a characteristic carbon- and oxygen-isotope pattern that allows for correlation with pelagic composite reference curves Vocontian and Umbria Marche basins.

Calibrated by means of foraminiferal biostratigraphy and rudist bivalve strontium-isotope stratigraphy, the section serves as local chemostratigraphic shallow-water reference for the Barremian to Cenomanian.

The m thick Monte La Costa section comprises predominantly coarse grained biostromal and often strongly cemented shelf margin deposits. Although benthic foraminifera are scarce and the carbonates evidently suffered strong diagenetic alteration, high-resolution rudist shell strontium-isotope stratigraphy in combination with superimposed carbon-isotope trends and biological-lithological changes e.

At both localities, chemostratigraphy indicates a major gap covering large parts of the Lower and middle Cenomanian. After having considerably improved the stratigraphic resolution of the studied sections, selected best-preserved rudist shells are going. Causal agents of Fusarium head blight of durum wheat Triticum durum Desf. Durum wheat samples harvested in central Italy Umbria were analyzed to: evaluate the occurrence of the fungal community in the grains, molecularly identify the Fusarium spp.

The Fusarium genus was one of the main components of the durum wheat fungal community. In vitro characterization of secondary metabolite biosynthesis was conducted for a wide spectrum of substances, showing the mycotoxigenic potential of the species complex.

Production of aurofusarin, butenolide, gibepyrone D, fusarin C, apicidin was also reported for the analyzed strains. Characterisation and reproduction of yellow pigments used in central Italy for decorating ceramics during Renaissance. This study presents the characterisation of prototypical yellow pigments used during the Renaissance period in Italy and the successful reproduction of homologous materials in accordance with the ancient recipes.

Moreover, a large number of yellow decorative layers of Sicilian ceramic artefacts dated back from 13th to the 19th century have been selected and the main chemical, structural and minero-petrografic features have been studied by X-ray diffraction, optical microscopy and scanning electron microscopy-energy dispersive spectrometry. These results have been compared with literature data of some yellow decorations of Renaissance ceramics made in central Italy.

Comparative arguments have shown some intriguing differences that are indicators of both technological transfer processes between central and southern Italy as well as of some local implementations likely due to specific raw materials locally available. Geomorphic evidence of active faults growth in the Norcia seismic area central Apennines, Italy.

Moreover the sub-basins represent the surface evidence of traits belonging to the Norcia seismogenic structure, which have repeatedly caused earthquakes in the past, thus determining similar geological, structural and morphostructural features within the wider Norcia area, without causing the whole structure to rupture. The size of these sub-basins and, thus, the size of the relevant seismogenic segments, allows to calculate a maximum magnitude for the three sub-basins and for the seismogenic area as a whole.

References Aringoli D. Materazzi M, Nibbi L. Morphotectonic characterization of the quaternary intermontane basins in the Umbria -Marche Apennines Italy. Lincei 25 Suppl 2 , SS DOI Tondi, E. Journal of Geodynamics, 36, Two cases of acute chest discomfort and the Central Italy earthquake. We present the cases of two postmenopausal women presenting to our emergency department with acute chest discomfort soon after the Central Italy earthquake.

Different diagnoses were made in the two patients. The role of the earthquake as a stressful event triggering diverse chest pain syndromes is discussed. Ethnobotanical remarks on Central and Southern Italy.

Background The present paper is a brief survey on the ethnobotanical works published by the Authors since , concerning the research carried out in some southern and central Italian regions.

Before Roman domination these territories were first inhabited by local people, while the southern areas were colonized by the Greeks. These different cultural contributions left certain traces, both in the toponyms and in the vernacular names of the plants and, more generally, in the culture as a whole. Methods Field data were collected through open interviews, mainly of farmers, shepherds and elderly people, born or living in these areas for a long time.

Voucher specimens of collected plants are preserved in the respective herbaria of the Authors and in the herbarium of “Roma Tre” University. Important contributions have been made by several students native to the areas under consideration. A comparative analysis with local specific ethnobotanical literature was carried out.

Results The paper reports several examples concerning human and veterinary popular medicine and in addition some anti-parasitic, nutraceutic, dye and miscellaneous uses are also described.

Moreover vernacular names and toponyms are cited. Eight regions of central and southern Italy particularly Latium, Abruzzo, Marche and Basilicata were investigated and the data obtained are presented in 32 papers. Most of the species of ethnobotanical interest have been listed in Latium species , Marche and Abruzzo The paper also highlights particularly interesting aspects or uses not previously described in the specific ethnobotanical literature.

Conclusion Phyto-therapy in central and southern Italy is nowadays practised by a few elderly people who resort to medicinal plants only for mild complaints on the contrary food uses are still commonly practised.

Nowadays therapeutic uses, unlike in the past, are less closely or not at all linked to ritual aspects. Several plants deserve to be taken. A nearly complete and undisturbed Miocene carbonate sequence is present in the easternmost part of the Umbria -Marche basin, Italy , which is ideal for detailed and integrated stratigraphic investigations of the Miocene Epoch.

In this study, we were trying to obtain evidence for the presence or absence of distal ejecta from the 15 Ma Ries impact structure in southern Germany, located about km to the north-northwest of the Umbria -Marche basin.

The first step is to find coeval strata in the Umbria -Marche sequence. Together with detailed geologic and stratigraphic information about the Aldo Level, the resulting age can be used confidentially to calibrate the Langhian stage. Besides providing new constraints on Miocene geochronology, this age can now be used for impact stratigraphic studies.

To directly correlate the biotite ages of the La Vedova section with rocks from the Ries impact event, Ries impact glass was also analyzed and found to be coeval. Although unrelated to this impact event, the biotite-rich clay layer should help in the search for evidence of distal ejecta related to the Ries crater. We present a seismic source model for Central Europe Belgium, Germany, Switzerland, and Austria and Italy , as part of an overall seismic risk and loss modeling project for this region.

A separate presentation at this conference discusses the probabilistic seismic hazard and risk assessment Williams et al. Where available we adopt regional consensus models and adjusts these to fit our format, otherwise we develop our own model.

Our seismic source model covers the whole region under consideration and consists of the following components: 1. A subduction zone environment in Calabria, SE Italy , with interface events between the Eurasian and African plates and intraslab events within the subducting slab. The subduction zone interface is parameterized as a set of dipping area sources that follow the geometry of the surface of the subducting plate, whereas intraslab events are modeled as plane sources at depth; 2.

The main normal faults in the upper crust along the Apennines mountain range, in Calabria and Central Italy. Dipping faults and sub- vertical faults are parameterized as dipping plane and line sources, respectively; 3. The Upper and Lower Rhine Graben regime that runs from northern Italy into eastern Belgium, parameterized as a combination of dipping plane and line sources, and finally 4. Background seismicity, parameterized as area sources. The fault model is based on slip rates using characteristic recurrence.

The modeling of background and subduction zone seismicity is based on a compilation of several national and regional historic seismic catalogs using a Gutenberg-Richter recurrence model.

Merging the catalogs encompasses the deletion of double, fake and very old events and the application of a declustering algorithm Reasenberg, The resulting catalog contains a little over events, has an average b-value of Abstract We present the cases of two postmenopausal women presenting to our emergency department with acute chest discomfort soon after the Central Italy earthquake. Cross-correlation analysis of seismic events in Central -Northern Italy : insights from the geochemical monitoring network of Tuscany.

Since late , a geochemical monitoring network is operating in Tuscany, Central Italy , to collect data and possibly identify geochemical anomalies that characteristically occur before regionally significant i. The network currently consists of 6 stations located in areas already investigated in detail for their geological setting, hydrogeological and geochemical background and boundary conditions. All these stations are equipped for remote, continuous monitoring of selected physicochemical parameters temperature, pH, redox potential, electrical conductivity , and dissolved concentrations of CO2 and CH4.

Additional information are obtained through in situ discrete monitoring. Field surveys are periodically performed to guarantee maintenance and performance control of the sensors of the automatic stations, and to collect water samples for the determination of the chemical and stable isotope composition of all the springs investigated for seismic precursors.

Geochemical continuous signals are numerically processed to remove outliers, monitoring errors and aseismic effects from seasonal and climatic fluctuations. The elaboration of smoothed, long-term time series more than data available today for each station allows for a relatively accurate definition of geochemical background values. Geochemical values out of the two-sigma relative standard deviation domain are inspected as possible indicators of physicochemical changes related to regional seismic activity.

Cross-correlation analysis indicates an apparent relationship between the most important seismic. An assessment of the feasibility of the use of satellite-only rainfall estimates for the hydrological monitoring in central Italy. The need for accurate distributed hydrological modelling has constantly increased in last years for several purposes: agricultural applications, water resources management, hydrological balance at watershed scale, floods forecast.

The main input for the hydrological numerical models is rainfall data that present, at the same time, a large availability of measures in gauged regions, with respect to other micro-meteorological variables and the most complex spatial patterns.

While also in presence of densely gauged watersheds the spatial interpolation of the rainfall is a non-trivial problem, due to the spatial intermittence of the variable especially at finer temporal scales , ungauged regions need an alternative source of rainfall data in order to perform the hydrological modelling.

Such source can be constituted by the satellite-estimated rainfall fields, with reference to both geostationary and polar-orbit platforms. In this work the rainfall product obtained by the Aqua-AIRS sensor were used in order to assess the feasibility of the use of satellite-based rainfall as input for distributed hydrological modelling.

In particular three experiments were carried on: a hydrological simulation with the use of rain-gauges data, b simulation with the use of satellite-only rainfall estimates, c simulation with the combined use of the two sources of data in order to obtain an optimal estimate of the actual rainfall fields.

The domain of the study was the central Italy. Several critical events occurred in the area were analyzed. A discussion of the results is provided. Case ascertainment was based on the presence of a primary nodular lesion in the lung and cytological or histological documentation of cancer from a primary or metastatic site. Outcome measures Sensitivity, specificity and positive predictive value PPV for Results cases and 94 non-cases were randomly selected from each database and the corresponding medical charts were reviewed.

Most of the diagnoses for lung cancer were performed in medical departments. True positive rates were high for all the three units.

Conclusions Case ascertainment for lung cancer based on imaging or endoscopy associated with histological examination yielded an excellent sensitivity in all the three administrative databases. We defined a seismogenic source model for central Italy and computed the relative forecast scenario, in order to submit the results to the CSEP Collaboratory for the study of Earthquake Predictability, www.

The goal of CSEP project is developing a virtual, distributed laboratory that supports a wide range of scientific prediction experiments in multiple regional or global natural laboratories, and Italy is the first region in Europe for which fully prospective testing is planned. This case-referent study evaluated cancer risks among farmers in central Italy.

Farmers had a decreased risk of lung and bladder cancer and melanoma and nonsignificant excess risks for stomach, rectal, kidney, and nonmelanoma skin cancer. Possible relationships emerged between specific crops and cancer: fruit and colon and bladder cancer, wheat and prostate cancer, olives and kidney cancer, and potato and kidney cancer. The results regarding stomach, pancreatic, lung, bladder, and prostate cancer and melanoma congrue with earlier results.

The kidney cancer excess, the association of colon and bladder cancer with orchard farming, and the excess of rectal cancer among licensed farmers are new and unexpected findings. Volatile components of horsetail Hippuris vulgaris L.

Hippuris vulgaris, also known as horsetail or marestail, is a freshwater macrophyte occurring in lakes, rivers, ponds and marshes. In the present study, we analysed for the first time the volatile composition of H. For the purpose, the essential oil was obtained by hydrodistillation and analysed by GC-MS. The chemical composition was dominated by aliphatic compounds such as fatty acids These compounds are here proposed as chemotaxonomic markers of the species.

Hydrogeochemical response of groundwater springs during central Italy earthquakes 24 August and October Co-seismic hydrological and chemical response at groundwater springs following strong earthquakes is a significant concern in the Apennines, a region in central Italy characterized by regional karstic groundwater systems interacting with active normal faults capable of producing Mw 6. These aquifers also provide water supply to major metropolitan areas in the region.

On August 24, , a Mw 6. The epicenter of the event was located at the segment boundary between the Mt. Vettore and Mt. Laga faults. On October 26, and on October 30, , three other big shocks Mw 5. Immediately after Aug. Thermal springs connected with deep groundwater flowpaths were also sampled.

These springs, sampled previously in and , provide some pre-earthquake data. Moreover, we sampled 4 springs along the Mt. Vettore fault system: 3 small springs at Forca di Presta, close to the trace of the earthquake surface ruptures, and two in Castel Sant’Angelo sul Nera. The latter are feeding the Nera aqueduct and the Nerea S.

The aim of this study is to evaluate the strong earthquake sequence effects on the hydrochemistry and flow paths of groundwater from different aquifer settings based on analysis before and after seismic events. The comparison between the responses of springs ca.

Epiphytic lichens as sentinels for heavy metal pollution at forest ecosystems central Italy. The results of a study using epiphytic lichens Parmelia caperata as sentinels for heavy metal deposition at six selected forest ecosystems of central Italy are reported.

The woods investigated are characterized by holm oak Quercus ilex , turkey oak Quercus cerris and beech Fagus sylvatica and represent the typical forest ecosystems of central Italy at low, medium and high elevations, respectively. The results showed that levels of heavy metals in lichens were relatively low and consequently no risk of heavy metal air pollution is expected for the six forest ecosystems investigated.

However, for two of them there are indications of a potential risk: the beech forest of Vallombrosa showed signs of contamination by Pb as a consequence of vehicle traffic due to the rather high touristic pressure in the area, and the holm oak forest of Cala Violina showed transboundary pollution by Mn, Cr and Ni originating from the steel industry in Piombino.

Epiphytic lichens proved to be very effective as an early warning system to detect signs of a changing environment at forest ecosystems. Microstructural investigations on carbonate fault core rocks in active extensional fault zones from the central Apennines Italy. The study of the microstructural and petrophysical evolution of cataclasites and gouges has a fundamental impact on both hydraulic and frictional properties of fault zones.

In the last decades, growing attention has been payed to the characterization of carbonate fault core rocks due to the nucleation and propagation of coseismic ruptures in carbonate successions e. Among several physical parameters, grain size and shape in fault core rocks are expected to control the way of sliding along the slip surfaces in active fault zones, thus influencing the propagation of coseismic ruptures during earthquakes.

Nevertheless, the role of grain size and shape distribution evolution in controlling the weakening or strengthening behavior in seismogenic fault zones is still not fully understood also because a comprehensive database from natural fault cores is still missing. In this contribution, we present a preliminary study of seismogenic extensional fault zones in Central Apennines by combining detailed filed mapping with grain size and microstructural analysis of fault core rocks.

Field mapping was aimed to describe the structural architecture of fault systems and the along-strike fault rock distribution and fracturing variations. In the laboratory we used a Malvern Mastersizer granulometer to obtain a precise grain size characterization of loose fault rocks combined with sieving for coarser size classes. In addition, we employed image analysis on thin sections to quantify the grain shape and size in cemented fault core rocks. The studied fault zones consist of an up to m-thick fault core where most of slip is accommodated, surrounded by a tens-of-meters wide fractured damage zone.

Fault core rocks consist of 1 loose to partially cemented breccias characterized by different grain size from several cm up to mm and variable grain shape from very angular to sub. Design A diagnostic accuracy study comparing melanoma ICDCM codes index test with medical chart reference standard. Case ascertainment was based on neoplastic lesion of the skin and a histological diagnosis from a primary or metastatic site positive for melanoma.

Participants , and cases subjects with melanoma were randomly selected from Umbria , NA and FVG, respectively; 94 non-cases subjects without melanoma were randomly selected from each unit. The mean age of the patients ranged from 60 to 61 years. Most of the diagnoses were performed in surgical departments.

Conclusions The case definition for melanoma based on clinical or instrumental diagnosis, confirmed by histological examination, showed excellent sensitivities and good specificities in the three operative units. Administrative databases from the three operative units can be used for epidemiological and outcome research of melanoma. A diagnostic accuracy study comparing melanoma ICDCM codes index test with medical chart reference standard. The most common melanoma subtype was malignant melanoma of skin of trunk, except scrotum ICDCM code: The case definition for melanoma based on clinical or instrumental diagnosis, confirmed by histological examination, showed excellent sensitivities and good specificities in the three operative units.

No commercial use is permitted unless otherwise expressly granted. The L’Aquila earthquake Mw 6. Since then, guidelines for microzonation were drew up that take into consideration the problem of surface faulting in Italy , and laying the bases for future regulations about related hazard, similarly to other countries e.

More specific guidelines on the management of areas affected by active and capable faults i. As such, the guidelines arise the problem of the time interval and general operational criteria to asses fault capability for the Italian territory. As for the chronology, the review of the international literature and regulatory allowed Galadini et al.

As for the operational criteria, the detailed analysis of the large amount of works dealing with active faulting in Italy shows that investigations exclusively based on surface morphological features e. A test area for which active and capable faults can be first mapped based on such a classical but still effective methodological approach can be the central Apennines.

Reference Galadini F. The activation of Late Quaternary faults in the Central Apennines Italy could generate earthquakes with magnitude of about 6. The collection of radar echoes in three dimensions allows to map both the vertical and lateral continuity of shallow geometries of the fault zone Fz , imaging features with high resolution, ranging from few metres to centimetres and therefore imaging also local variations at the microscale.

Several geophysical markers of faulting, already highlighted on this site, have been taken as reference to plan the 3-D survey. In this paper, we provide the first 3-D subsurface imaging of an active shallow fault belonging to the Umbria -Marche Apennine highlighting the subsurface fault geometry and the stratigraphic sequence up to a depth of about 5 m.

From our data, geophysical faulting signatures are clearly visible in three dimensions: diffraction hyperbolas, truncations of layers, local attenuated zones and varying dip of the layers have been detected within the Fz. The interpretation of the 3-D data set provided qualitative and quantitative geological information in addition to the fault location.

Chemical analysis of essential oils from different parts of Ferula communis L. Ferula communis is a showy herbaceous plant typical of the Mediterranean area where it is used as a traditional medicine. The plant is a source of bioactive compounds such as daucane sesquiterpenes and prenylated coumarins.

In Italy , most of phytochemical studies focused on Sardinian populations where poisonous and nonpoisonous chemotypes were found, while investigations on peninsular populations are scarce. In this work, we report the chemical characterisation of the essential oils obtained from different parts of F. The volatile profiles detected did not allow to classify the investigated central Italy population into the poisonous and nonpoisonous chemotypes previously described in Sardinia.

First evaluation of endotoxins in veterinary autogenous vaccines produced in Italy by LAL assay. Endotoxin contamination is a serious concern for manufacturers of biological products and vaccines in terms of not only quality but also safety parameters. According to the 3Rs principles Replace, Reduce, Refine , which aim to progressively reduce animal use in the quality control process, we tested the vaccines obtained from gram-negative bacteria and adjuvants by the limulus amebocyte lysate LAL assay.

The results revealed low endotoxin concentrations compared to available data in the literature and represent the first report of the application of the 3Rs principles to veterinary autogenous vaccines production in Italy. Published by Elsevier Ltd. Towards the application of seismogeodesy in central Italy : a case study for the August 24 Mw 6.

Dense strong motion and high-rate Global Navigation Satellite Systems GNSS networks have been deployed in central Italy for rapid seismic source determination and corresponding hazard mitigation.

Different from previous studies for the consistency between two kinds of sensor at collocated stations, here we focus on the combination of high-rate GNSS displacement waveforms with collocated seismic strong motion accelerators, and investigate its application to image rupture history. Taking the August 24 Mw 6. Our results reveal that strong motion data alone can overestimate the magnitude and mismatch the GNSS observations, while 1 Hz sampling rate GNSS is insufficient and the displacement is too noisy to depict rupture process.

By contrast, seismogeodetic data enhances temporal resolution and maintains the static offsets that provide vital constraint to the reliable estimation of earthquake magnitude. The obtained model is close to the jointly inverted one. Our work demonstrates the unique usefulness of seismogeodesy for fast seismic hazard response.

Morphology and evolution of sulphuric acid caves in South Italy. D’Angeli, Ilenia M. Sulphuric acid speleogenesis SAS related to the upwelling of acid water enriched in H2S and CO2 represents an unusual way of cave development. Since meteoric infiltration waters are not necessarily involved in speleogenesis, caves can form without the typical associated karst expressions i. The main mechanism of sulphuric acid dissolution is the oxidation of H2S Jones et al.

In these conditions, carbonate dissolution associated with gypsum replacement, is generally believed to be faster than the normal epigenic one De Waele et al. Other preliminary studies have been carried out in Calabria Galdenzi, and Sicily De Waele et al. Using both limestone tablet weight loss Galdenzi et al. Geomorphological observations, landscape analysis using GIS tools, and the analysis of gypsum and other secondary minerals alunite and jarosite stable isotopes and dating will help to reconstruct the speleogenetic stages of cave formation.

Preliminary microbiological analysis will determine the microbial diversity and ecology in the biofilms. References Engel S. In order to study the main features of the MRA reform we used a methodological approach based on three steps: i first we applied a desk analysis to sketch the…. Relationship between work-related accidents and hot weather conditions in Tuscany central Italy. Nowadays, no studies have been published on the relationship between meteorological conditions and work-related mortality and morbidity in Italy.

The aim of this study was to evaluate the relationship between hot weather conditions and hospital admissions due to work-related accidents in Tuscany central Italy over the period Apparent temperature AT values were calculated to evaluate human weather discomfort due to hot conditions and then tested for work accident differences using non-parametric procedures. Present findings showed that hot weather conditions might represent a risk factor for work-related accidents in Italy during summer.

In particular early warming days during June, characterized by heat discomfort, are less tolerated by workers than warming days of the following summer months. The peak of work-related accidents occurred on days characterized by high, but not extreme, thermal conditions. Workers maybe change their behaviour when heat stress increases, reducing risks by adopting preventive measures.

Results suggested that days with an average daytime AT value ranged between Chemical weathering of palaeosols from the Lower Palaeolithic site of Valle Giumentina, central Italy. The evolution of the major and trace element composition suggests that the palaeosols were mainly formed by in situ weathering of the parent material. The major phases of soil weathering occurred during the MIS 13 interglacial period S8 and S6 as well as during episodes of rapid environmental change associated with millennial climatic oscillations during the MIS 14 and 12 glaciations S11 and S2, respectively.

Although global forcing such as orbital variations, solar radiation, and greenhouse gas concentrations may have influenced the pedogenic processes, the volcanism in central Italy , climate change in the central Mediterranean, and tectono-sedimentary evolution of the Valle Giumentina basin also impacted and triggered the formation of most palaeosols, which provided subsistence resources for the Lower Palaeolithic human communities.

This study highlights the importance of having high-resolution palaeoenvironmental records with accurate chronology as close as possible to archaeological sites to study human-environment interactions.

Combining historical and geomorphological information to investigate earthquake induced landslides. Landslides are caused by many different triggers, including earthquakes. In Italy , a detailed new generation catalogue of information on historical earthquakes for the period B. The catalogue lists earthquakes and provides information on a total of about mass-movements triggered by seismic events. The information on earthquake-induced landslides listed in the catalogue was obtained through the careful scrutiny of historical documents and chronicles, but was rarely checked in the field.

We report on an attempt to combine the available historical information on landslides caused by earthquakes with standard geomorphological techniques, including the interpretation of aerial photographs and field surveys, to better determine the location, type and distribution of seismically induced historical slope failures.

We present four examples in the Central Apennines. The landslide is the oldest known earthquake-induced slope failure in Italy. The landslides were subsequently largely modified by mass movements occurred on making the recognition of the original seismically induced failures difficult, if not impossible. In the third example we present the geographical distribution of the available information on landslide events triggered by 8 earthquakes in Central Valnerina, in the period to A comparison with the location of landslides triggered by the September-October Umbria -Marche earthquake sequence is presented.

The fourth example describes the geographical distribution of the available. Six components observations of local earthquakes during the Central Italy seismic sequence. For many years the seismological community has looked for a reliable, sensitive, broadband three-component portable rotational sensor. In this preliminary study, we show the possibility of measuring and extracting relevant seismological information from local earthquakes.

We employ portable three-component rotational sensors, insensitive to translations, which operate on optical interferometry principles Sagnac effect. Multiple sensors recording redundantly add significance to the measurements. We present here the six-component observations, with analysis of rotational three redundant components and translational three components ground motions, generated by earthquakes at local distances.

For each seismic event, we compare coherence between rotational sensors and estimate a back azimuth consistent with theoretical values. We also estimate Love and Rayleigh wave phase velocities in the 5 to 10 Hz frequency range. Source characteristics of small earthquakes nucleating on the Alto Tiberina fault system central Italy. The ATF has been imaged by analyzing the active source seismic reflection profiles, and the instrumentally recorded persistent background seismicity.

The present study is an attempt to separate the contributions of source, site, and crustal attenuation, in order to focus on the mechanics of the seismic sources on the ATF, as well on the synthetic and the antithetic structures within the ATF hanging-wall i. Colfiorito fault, Gubbio fault and Umbria Valley fault. In order to compute source spectra, we perform a set of regressions over the seismograms of small earthquakes Orain, R.

The palaeobotanical record of early Palaeolithic sites from Western Europe indicates that hominins settled in different kinds of environments. During the “mid-Pleistocene transition MPT “, from about 1 to 0. Starting from the MPT the more favourable climate of central and southern Italy provided propitious environmental conditions for long-term human occupations even during the glacial times.

In fact, the human strategy of territory occupation was certainly driven by the availabilities of resources. Prehistoric sites such as Notarchirico ca. Faunal remains indicate that human subsistence behaviours benefited from a diversity of exploitable ecosystems, from semi-open to closed environments.

In central and southern Italy , several palynological records have already illustrated the regional- and local-scale vegetation dynamic trends. During the Middle Pleistocene climate cycles, mixed mesophytic forests developed during the interglacial periods and withdrew in response to increasing aridity during the glacial episodes.

In this basin the persistence of high edaphic humidity, even during the glacial phases, could have favoured the establishment of a refuge area for the arboreal flora and provided subsistence resources for the animal and hominin communities during the Middle. Hominin responses to environmental changes during the Middle Pleistocene in Central and Southern Italy.

Starting from the MPT the more favorable climate of central and southern Italy provided propitious environmental conditions for long-term human occupations even during the glacial times. Faunal remains indicate that human subsistence behaviors benefited of a diversity of exploitable ecosystems, from semi-open to closed environments. In central and southern Italy , several palynological records have already illustrated the regional and local scale vegetation dynamic trends.

New pollen data from the Boiano basin Molise, Italy , attest to the evolution of vegetation and climate between OIS 13 and 9 ca. In this basin, the persistence of high edaphic humidity, even during the glacial phases, could have favored the establishment of a refuge area for the arboreal flora and provided subsistence resources for the animal and hominin communities during the Middle.

During a landslide triggering event, the tens to thousands of landslides resulting from the trigger e. Here, we present initial results from a semi-stochastic model we are developing to evaluate the probability of landslides intersecting a road network and the network-accessibility implications of this across a region.

This was performed in the open source GRASS GIS software, where we took ‘model’ landslides and dropped them on a 79 km2 test area region in Collazzone, Umbria , Central Italy , with a given road network major and minor roads, km in length and already determined landslide susceptibilities. Landslide areas AL were randomly selected from a three-parameter inverse gamma probability density function, consisting of a power-law decay of about After a point was accepted, it was assigned a landslide area AL and length to width ratio.

Landslide intersections with roads were then assessed and indices such as the location, number and size of road blockage recorded. Initial results show that for a landslide triggering event of 1 landslide km-2 over a 79 km2 region with km of road, the number of road blockages. The main goal of the EUROBIS is to change the pendency of slope of the actual trend towards the increase in the yearly rates of childhood overweight and obesity in Umbria and to improve healthy lifestyles of children and their parents.

The aims of the Italian EUROBIS study are: 1 a community-based intervention program CBP carrying out activities in all primary schools of the Umbria Region and family settings as first step, to reverse the current obesity trend on a long-term basis, and 2 a clinical care program for childhood and adolescent by C. The community-based intervention and clinical trial provide an innovative valuable model to address the childhood obesity prevention and treatment in Italy. Vaccine storage in the community: a study in central Italy.

Maintaining the vaccine cold chain is an essential part of a successful immunization programme, but in developed countries faulty procedures may occur more commonly than is generally believed. A survey was conducted in a health district in central Italy to assess the methods of vaccine transportation and storage. Of 52 primary vaccination offices inspected, 39 None of the seven main offices selected for monitoring had a maximum and minimum thermometer and none monitored the internal temperature of the refrigerator.

Moreover, other faulty procedures, such as the storage of food and laboratory specimens in vaccine refrigerators and the storage of vaccines on refrigerator door shelves, indicated that the knowledge and practice of vaccine storage and handling were often inadequate. Fishes and their parasites in the water district of Massaciuccoli Tuscany, Central Italy. This study has been conducted in the district of Massaciuccoli lake, marsh and reclaimed areas with drainage channels in Tuscany region Central Italy.

The aim of the research was to detect the presence of parasites in fishes, in particular of Opisthorchis felineus, which causes an important zoonosis. Between , the health status of fishes was monitored, morphometric characteristics were determined, and parasites were searched for and identified. Of the examined fishes, were free of parasites while were infected, among them 91 presented multiple infections.

Opisthorchis felineus was not found in any of the examined fishes. Multiple source components in gas manifestations from north- central Italy. Gas manifestations in north- central Italy consist of CO2-rich gases with minor N2-rich emissions and discharge either along with thermal springs or into cold and stagnant waters.

The variable composition of ‘thermal’ gases is caused by differences in the thermal regime and lithology of the ascent paths, where there is mixing of gases from multiple sources, such as N2-rich atmospheric and deep CO2-rich metamorphic end-members. Elevated concentrations of CH4 and H2 in these gases are generally related to the presence of active geothermal systems at shallow depth, such as the Larderello-Travale field in Tuscany.

C hydrothermal systems. In some cases, they can be scrubbed or oxidized especially H2 , while the residual rising gas becomes indirectly enriched in N2 and CO2. Carbon dioxide is also enriched in some discharged gases because it is produced at shallow depth in lower temperature conditions central Italy. A significant component of mantle 3He is only found in the geothermal areas of Larderello where the large regional thermal anomaly. Ethnophytotherapeutical research in the high Molise region Central -Southern Italy.

Background In the years — research was carried out concerning ethno-medicine in the high Molise central – southern Italy , a region that has been the object of very little investigation from the ethnobotanical point of view.

Upper Molise is a continuation of the mountain profiles of the Abruzzi Appenines: a series of hills, steep slopes and deep fluvial valleys making communications difficult. Primordial traditions e. Methods Field data were collected through open interviews in the field.

These were conducted on both an individual and group level, fresh plants gathered from surrounding areas being displayed. In other cases, individual interviews were conducted by accompanying the person involved to the places where they perform their activities for example, in the woods with woodcutters, kitchen gardens and fields with housewives, pastures with shepherds, etc. In total 54 individuals were interviewed. Results Data of 70 taxa belonging to 39 families were gathered.

Among the species, 64 are used in human therapy, 5 as insect repellents, 11 in veterinary medicine, 1 to keep eggs and cheeses and 4 for magic purposes. The most important findings in ethno-medicine relate to the lichen Lobaria pulmonaria L. Also worthy of note, given the isolation of the area, is the number of plants used to protect foodstuffs from parasites, among which Allium sativum L.

Conclusion The research revealed a deep-rooted and widespread habit of husbanding the family’s resources. Whilst isolation and snowfalls contributed to the widespread knowledge of means. Persistency of rupture directivity in moderate-magnitude earthquakes in Italy : Implications for seismic hazard. A simple method based on the EGF deconvolution in the frequency domain is applied to detect the occurrence of unilateral ruptures in recent damaging earthquakes in Italy.

The spectral ratio between event pairs with different magnitudes at individual stations shows large azimuthal variations above corner frequency when the target event is affected by source directivity and the EGF is not or vice versa. The analysis is applied to seismograms and accelerograms recorded during the seismic sequence following the 20 May , Mw 5.

Events of each seismic sequence are selected as having consistent focal mechanisms, and the station selection obeys to the constraint of a similar source-to-receiver path for the event pairs. The analyzed data set of L’Aquila consists of broad-band seismograms relative to 69 normal-faulting earthquakes 3. Also the Mw 5. According to the reconstructed geometry of the trust-fault plane, the inferred directivity direction suggests top-down rupture propagation. The analysis over the Emilia aftershock sequence is in progress.

The third seismic sequence, dated , occurred in the northern Apennines and, similarly. Environmental radioactivity in four national parks of the Abruzzo region central Italy. Caporale’ in Teramo is conducting radioecological surveys in the Abruzzo region Central Italy , to acquire knowledge on the geochemical and biological mobility of radionuclides derived from the Chernobyl accident.

To this end, samples of grasses, fungi, mosses and soils were collected in four national parks Sirente-Velino, Abruzzo Lazio and Molise, the Gran Sasso and the park of Monti della Laga and Maiella. The results show that the Chernobyl fallout is still detectable in the samples collected in the four parks but the Cs concentrations are present in the semi-natural environments in quantities that do not create concerns from a radiological point of view.

Assessment of local seismic response of the Stracciacappa maar Central Italy. In this work, we face the definition of a subsoil model aimed at the local seismic response assessment of the Stracciacappa maar Sabatini Volcanic District, central Italy e. The pyroclastic succession of Stracciacappa records two main hydromagmatic eruptive phases ended about 0. The preserved crater, with a diameter of about meters and a crater floor of about m, hosted a lake until it was drained in AD New geological map and cross sections illustrate the complex geometric relationships between the thick pyroclastic surge succession, showing diffuse sandwave structures, and even meter-sized lava ballistic.

A composite interdigitation between lacustrine and epiclastic debris sediments fills the crater floor. A continuous coring borehole was drilled inside the crater, 45 meters deep from the wellhead, with sampling of undisturbed samples. This Vs profile was then extended at higher depths by using the results of four 2D seismic passive arrays. Finally, three electrical resistivity tomography tests, with a total length of about meters, were carried out with the purpose of constraining the subsoil model.

Regarding the non linear. The Central Italy Earthquake: an Overview. The M6 central Italy earthquake occurred on the seismic backbone of the Italy , just in the middle of the highest hazard belt. The shock hit suddenly during the night of August 24, when people were asleep; no foreshocks occurred before the main event.

The earthquake ruptured from 10 km to the surface, and produced a more than 17, aftershocks Oct. It is geologically very similar to previous recent events of the Apennines. Both the L’Aquila earthquake to the south and the Colfiorito to the north, were characterized by the activation of adjacent fault segments.

Despite its magnitude and the well known seismic hazard of the region, the earthquake produced extensive damage and fatalities.

The town of Amatrice, that paid the highest toll, was classified in zone 1 the highest since , but the buildings in this and other villages revealed highly vulnerable. In contrast, in the town of Norcia, that also experienced strong ground shaking, no collapses occurred, most likely due to the retrofitting carried out after an earthquake in The first field teams reached the epicentral area at 7 am with the portable seismic stations installed to monitor the aftershocks; other teams followed to map surface faults, damage, to measure GPS sites, to install instruments for site response studies, and so on.

DPC , the media and the web. Several tens of reports and updates have been delivered in the first month of the sequence to DPC. Also due to the controversial situation arisen from the L’Aquila earthquake and trials, particular attention was given to the communication: continuous and timely information has been released to. These phenomena generally present evident geomorphological markers e.

To understand which is the most influential factor in DSGSDs’ activity is rarely an easy task because this can vary from case to case.

This work illustrates the outcomes provided by a monitoring activity conducted along the Mt. Frascare slope Fiastra Lake, Marche region, Italy. The monitoring system is composed by 4 low cost GPS stations, based on single-frequency receivers, and 2 double-frequency GPS stations, aimed to cross-check the surface deformations measured by the two types of monitoring stations.

The 6 GPS stations have been operated in place starting from October grounded on the base of a geomorphological field survey of the investigated phenomenon. Two stations have been equipped with both receiver types in order to facilitate the comparison of the results.

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