The Evolution of Diet and Microhabitat Use in Pseudoboine Snakes

Reflecting their exceptional radiation, snakes occur in different habitats and microhabitats and are able to eat numerous types of prey. The availability of good and comprehensive phylogenies for different snake’s lineages together with natural history data provides an opportunity to explore how ecological traits diversified during their radiation. In the present study, we describe the diet and microhabitat variation (arboreal or non-arboreal) in the tribe Pseudoboini and explore how these traits evolved during the tribe’s diversification. We analyzed specimens deposited in scientific collections and gathered information on diet and microhabitat use available in the literature and provided by other researchers. We also mapped diet and microhabitat data onto a phylogeny of the tribe using the principle of parsimony. Pseudoboine snakes feed mainly on lizards and small mammals, and of the 22 species for which a minimum number of prey records was obtained, nine are diet generalists, six are lizard specialists, three are small mammal specialists, two are snake specialists, one is a lizard egg specialist, and one is a bird egg specialist. The highly diverse feeding habits of pseudoboines seem to have evolved mainly in the terminal taxa. Among those species that had enough microhabitat data (17 species), Drepanoides anomalus, Siphlophis cervinus, S. compressus, and S. pulcher frequently use the vegetation. Our results indicate that an increase in arboreality evolved several times during the diversification of the tribe, and that the Siphlophis clade seems to have maintained the high degree of arboreality from its ancestor. Species that frequently use vegetation are either lizard or lizard egg specialists, indicating that these habits might be associated in the evolution of pseudoboines.

Study of Italian isolates of Alternaria spp.: molecular and morphological characterization and pathogenesis on apple tree

It was decided to carry out a morphological and molecular characterization of the Italian Alternaria isolatescollected from apple , and evaluate their pathogenicity and subsequently combining the data collected. The strain collection (174 isolates) was constructed by collecting material (received from extension service personnel) between June and August of 2007, 2008, and 2009. A Preliminary bioassays were performed on detached plant materials (fruit and leaf wounded and unwounded), belonging to the Golden cultivar, with two different kind of inoculation (conidial suspension and conidial filtrate). Symptoms were monitored daily and a value of pathogenicity score (P.S.) was assigned on the basis of the diameter of the necrotic area that developed. On the basis of the bioassays, the number of isolates to undergo further molecular analysis was restricted to a representative set of single spore strains (44 strains). Morphological characteristics of the colony and sporulation pattern were determined according to previous systematic work on small-spored Alternaria spp. (Pryor and Michaelides, 2002 and Hong et al., 2006). Reference strains (Alternaria alternata, Alternaria tenuissima, Alternaria arborescens and four Japanese strains of Alternaria alternata mali pathotype), used in the study were kindly provided by Prof. Barry Pryor, who allows a open access to his own fungal collection. Molecular characterization was performed combining and comparing different data sets obtained from distinct molecular approach: 1) investigation of specific loci and 2) fingerprinting based on diverse randomly selected polymorphic sites of the genome. As concern the single locus analysis, it was chosen to sequence the EndoPG partial gene and three anonymous region (OPA1-3, OPA2- and OPa10-2). These markers has revealed a powerful tool in the latter systematic works on small-spored Alternaria spp. In fact, as reported in literature small-spored Alternaria taxonomy is complicated due to the inability to resolve evolutionary relationships among the taxa because of the lack of variability in the markers commonly used in fungi systematic. The three data set together provided the necessary variation to establish the phylogenetic relationships among the Italian isolates of Alternaria spp. On Italian strains these markers showed a variable number of informative sites (ranging from 7 for EndoPg to 85 for OPA1-3) and the parsimony analysis produced different tree topologies all concordant to define A. arborescens as a mophyletic clade. Fingerprinting analysis (nine ISSR primers and eight AFLP primers combination) led to the same result: a monophyleic A. arborescens clade and one clade containing both A. tenuissima and the A. alternata strains. This first attempt to characterize Italian Alternaria species recovered from apple produced concordant results with what was already described in a similar phylogenetic study on pistachio (Pryor and Michaelides, 2002), on walnut and hazelnut (Hong et al., 2006), apple (Kang et al., 2002) and citurus (Peever et al., 2004). Together with these studies, this research demonstrates that the three morphological groups are widely distributed and occupy similar ecological niches. Furthermore, this research suggest that these Alternaria species exhibit a similar infection pattern despite the taxonomic and pathogenic differences. The molecular characterization of the pathogens is a fundamental step to understanding the disease that is spreading in the apple orchards of the north Italy. At the beginning the causal agent was considered as Alteraria alternata (Marshall and Bertagnoll, 2006). Their preliminary studies purposed a pathogenic system related to the synthesis of toxins. Experimental data of our bioassays suggest an analogous hypothesis, considering that symptoms could be induced after inoculating plant material with solely the filtrate from pathogenic strains. Moreover, positive PCR reactions using AM-toxin gene specific primers, designed for identification of apple infecting Alternaria pathovar, led to a hypothesis that a host specific toxin (toxins) were involved. It remains an intriguing challenge to discover or not if the agent of the “Italian disease” is the same of the one previously typified as Alternaria mali, casual agent of the apple blotch disease.

Phylogeny and historical biogeography of the Australian Camphorosmeae (Chenopodiaceae)

Diese Studie befasst sich mit der Phylogenie und Biogeographie der australischen Camphorosmeae, die ein wichtiges Element der Flora arider Gebiete Australiens sind. Die molekularen Phylogenien wurden mit Hilfe Bayes’scher Statistik und „maximum likelihood”berechnet. Um das Alter der Gruppe und interner Linien abzuschätzen, wurden die Methoden „Nonparametric rate smoothing” und “penalized likelihood” benutzt. Morphologische Merkmale wurden nach Kriterien der Parsimonie auf den molekularen Baum aufgetragen. „Brooks parsimony analysis”, „cladistic analysis of distributions and endemism”, „dispersal-vicariance analysis”,„ancestral area analysis” und „weighted ancestral area analysis” wurden angewandt, um Abfolge und Richtungen der Ausbreitung der Gruppe in Australien zu analysieren.Von sieben getesteten Markern hatten nur die nukleären ETS und ITS genügend Variation für die phylogenetische Analyse der Camphorosmeae. Die plastidären Marker trnL-trnF spacer,trnP-psaJ spacer, rpS16 intron, rpL16 intron und trnS-trnG spacer zeigten kein ausreichendes phylogenetisches Signal. Die gefundenen phylogenetischen Hypothesen widersprechen der jetzigen Taxonomie der Gruppe. Neobassia, Threlkeldia, Osteocarpum und Enchylaena sollten den Gattungen Sclerolaena bzw. Maireana zugeordnet werden. Die kladistische Analyse der Fruchtanhängsel unterstützt die taxonomischen Ergebnisse der auf DNA basierenden Phylogenie. Allerdings hat die Behaarung, die bei anderen Gruppen der Chenopodiaceae als wichtiges taxonomisches Merkmal herangezogen wird, die Phylogenie nicht unterstützt. Vorfahren der heutigen Camphorosmeen sind im Miozän, vor ca. 8-14 Millionen Jahren, durch Fernausbreitung vermutlich aus Asien in Australien eingewandert. Anfängliche Diversifizierung fand während des späten Miozäns bis in das frühe Pliozän vor ca. 4-7 Millionen Jahren statt. Am Ende des Pliozäns existierten schon 45% - 72% der Abstammungslinien der jetzigen Camphorosmeen. Dies weist auf eine schnelle Ausbreitung hin. Das Alter stimmt mit dem Einsetzen der Aridisierung Australiens überein, und deutet darauf hin, dass die Ausbreitung der ariden Gebiete eine große Rolle bei der Diversifizierung der Gruppe spielte. Die Vorfahren der australischen Camphorosmeae scheinen die Südküste Australiens zuerst besiedeln zu haben. Dies geschah vor dem Einsetzen der Aridisierung des Kontinents. Die anschließende Ausbreitung erfolgte in verschiedene Richtungen und folgte der fortschreitenden Austrocknung im späten Tertiär und im ganzen Quartär. Durch ihre Anpassung an Trockenheit ist der Erfolg der Camphorosmeae in den ariden Gebieten zu erklären.Die Abwesenheit von klaren phylogenetischen und artspezifischen Signalen zwischen Arten der australischen Camphorosmeae ist auf das junge Alter und die schnelle Diversifizierung der Gruppe zurückzuführen, welche die Häufung von Mutationen und eine starke morphologische Differenzierung nicht zugelassen haben.

Phylogeny of Western Palaearctic long-eared bats (Mammalia, Chiroptera, Plecotus) - a molecular perspective

Die Phylogenie der Westpaläarktischen Langohren (Mammalia, Chiroptera, Plecotus) – eine molekulare Analyse Die Langohren stellen eine Fledermausgattung dar, die fast alle westpaläarktischen Habitate bist zum Polarkreis hin besiedeln und in vielerlei Hinsicht rätselhaft sind. In der Vergangenheit wurden zahlreiche Formen und Varietäten beschrieben. Trotzdem galt für lange Zeit, dass nur zwei Arten in Europa anerkannt wurden. Weitere Arten waren aus Nordafrika, den Kanaren und Asien bekannt, aber auch deren Artstatus wurde vielfach in Frage gestellt. In der vorliegenden Dissertation habe ich mittels molekularer Daten,der partiellen Sequenzierung mitochondrialer Gene (16S rRNA und ND1), sowie der mitochondrialen Kontrollregion, eine molekular Analyse der phylogenetischen Verwandtschaftsverhältnisse innerhalb und zwischen den Linien der westpaläarktischen Langohren durchgeführt. Die besten Substitutionsmodelle wurden berechnet und phylogenetische Bäume mit Hilfe vier verschiedener Methoden konstruiert: dem neighbor joining Verfahren (NJ), dem maximum likelihood Verfahren (ML), dem maximum parsimony Verfahren (MP) und dem Bayesian Verfahren. Sechs Linien der Langohren sind genetisch auf einem Artniveau differenziert: Plecotus auritus, P. austriacus, P. balensis, P. christii, P. sardus, und P. macrobullaris. Im Falle der Arten P. teneriffae, P. kolombatovici und P. begognae ist die alleinige Interpretation der genetischen Daten einzelner mitochondrialer Gene für eine Festlegung des taxonomischen Ranges nicht ausreichend. Ich beschreibe in dieser Dissertation drei neue Taxa: Plecotus sardus, P. kolombatovici gaisleri (=Plecotus teneriffae gaisleri, Benda et al. 2004) and P. macrobullaris alpinus [=Plecotus alpinus, Kiefer & Veith 2002). Morphologische Kennzeichen, insbesondere für die Erkennung im Feld, werden hier dargestellt. Drei der sieben Arten sind polytypisch: P. auritus (eine west- und ein osteuropäische Linie, eine sardische Linie und eine aktuell entdeckte kaukasische Linie, Plecotus kolombatovici (P. k. kolombatovici, P. k. gaisleri und P. k. ssp.) und P. macrobullaris (P. m. macrobullaris und P. m. alpinus). Die Verbreitungsgebiete der meisten Arten werden in dieser Arbeit erstmals ausschließlich anhand genetisch zugeordneter Tiere dargestellt.Die Untersuchung der ökologischen Einnischung der nun anerkannten Formen, insbesondere in Gebieten sympatrischer Verbreitung, bietet ein spannendes und lohnendes Feld für zukünftige Forschungen. Nicht zuletzt muss sich die Entdeckung eines beachtlichen Anteils kryptischer Diversität innerhalb der westpaläarktischen Langohren auch bei der Entwicklung spezieller Artenschutzkonzepte widerspiegeln.

Revision of Trogulidae Sundevall, 1833 (Arachnida: Opiliones)

The relationship and phylogeny of the western Palearctic harvestmen family Trogulidae is investigated. The traditional system of seven genera and approximately 40 species appeared to be artificially composed but a phylogenetic approach and a comprehensive revision has long been sought after. Species are poorly characterised due to their uniform morphology and species evaluation is furthermore complicated by the variability of the few characters used for species delineation. To meet these demands a molecular genetic analysis is accomplished using the nuclear 28S rRNA gene and the mitochondrial cytochrome b gene. This analysis incorporates most genera and species of Trogulidae as well as a comprehensive set of Nemastomatidae and Dicranolasmatidae as outgroup taxa. Phylogenetic results of Bayesian analysis, Maximum Parsimony, Maximum Likelihood and Neighbor Joining are compared with distributional data, morphological characters and results of canonical discriminant analysis of morphometric characters and general congruence of these data sets is shown. To demonstrate the applicability of this method the revision of two species-groups within Trogulus is set out in detail. The Trogulus hirtus species-group and the Trogulus coriziformis species-group are revised. The former is in the central and north-western Balkan Peninsula. T. tricarinatus ssp. hirtus is raised to species level and four new species are described (T. karamanorum [man.n.], T. melitensis [man.n.], T. pharensis [man.n]; T. thaleri [man.n.]). The Trogulus coriziformis species-group is confined to the western Mediterranean area. T. coriziformis, T. aquaticus are re-described, T. cristatus and T. lusitanicus are re-established and four species are described as new (T. balearicus, T. huberi, T. prietoi, T. pyrenaicus). In both species-groups two further cryptic species probably exist but were not described. The species groups are shown to represent different phylogenetic levels and this information is used for the revisional work on the genus Trogulus as well as for the generic system of Trogulidae. Family status of Dicranolasmatidae is rejected and Dicranolasma is shown to be best incorporated within Trogulidae. Calathocratus, Platybessobius and Trogulocratus appear to be polyphyletic and are best to be united within Calathocratus, the oldest name of this set. The cryptic diversity within Trogulidae, especially in Trogulus and the composed genus Calathocratus rates to 150-235% and is thereby remarkably high for a group of the generally well researched European fauna. Genetic features of the group such as heteroplasmy, the possibility of major gene rearrangements and usability of the cytochrome b gene for phylogenetic studies in Opiliones are outlined.

Hydraulic evaluation of the gyro electric wave energy converter

Motivation Thanks for a scholarship offered by ALma Mater Studiorum I could stay in Denmark for six months during which I could do physical tests on the device Gyro PTO at the Departmet of Civil Engineering of Aalborg University. Aim The goal of my thesis is an hydraulic evaluation of the device: Gyro PTO, a gyroscopic device for conversion of mechanical energy in ocean surface waves to electrical energy. The principle of the system is the application of the gyroscopic moment of flywheels equipped on a swing float excited by waves. The laboratory activities were carried out by: Morten Kramer, Jan Olsen, Irene Guaraldi, Morten Thøtt, Nikolaj Holk. The main purpose of the tests was to investigate the power absorption performance in irregular waves, but testing also included performance measures in regular waves and simple tests to get knowledge about characteristics of the device, which could facilitate the possibility of performing numerical simulations and optimizations. Methodology To generate the waves and measure the performance of the device a workstation was created in the laboratory. The workstation consist of four computers in each of wich there was a different program. Programs have been used : Awasys6, LabView, Wave lab, Motive optitrack, Matlab, Autocad Main Results Thanks to the obtained data with the tank testing was possible to make the process of wave analisys. We obtained significant wave height and period through a script Matlab and then the values of power produced, and energy efficiency of the device for two types of waves: regular and irregular. We also got results as: physical size, weight, inertia moments, hydrostatics, eigen periods, mooring stiffness, friction, hydrodynamic coefficients etc. We obtained significant parameters related to the prototype in the laboratory after which we scale up the results obtained for two future applications: one in Nissun Brending and in the North Sea. Conclusions The main conclusion on the testing is that more focus should be put into ensuring a stable and positive power output in a variety of wave conditions. In the irregular waves the power production was negative and therefore it does not make sense to scale up the results directly. The average measured capture width in the regular waves was 0.21 m. As the device width is 0.63 m this corresponds to a capture width ratio of: 0.21/0.63 * 100 = 33 %. Let’s assume that it is possible to get the device to produce as well in irregular waves under any wave conditions, and lets further assume that the yearly absorbed energy can be converted into electricity at a PTO-efficiency of 90 %. Under all those assumptions the results in table are found, i.e. a Nissum Bredning would produce 0.87 MWh/year and a North Sea device 85 MWh/year.

The evolution of antennal courtship in diplazontine parasitoid wasps (Hymenoptera, Ichneumonidae, Diplazontinae)

Background As predicted by theory, traits associated with reproduction often evolve at a comparatively high speed. This is especially the case for courtship behaviour which plays a central role in reproductive isolation. On the other hand, courtship behavioural traits often involve morphological and behavioural adaptations in both sexes; this suggests that their evolution might be under severe constraints, for instance irreversibility of character loss. Here, we use a recently proposed method to retrieve data on a peculiar courtship behavioural trait, i.e. antennal coiling, for 56 species of diplazontine parasitoid wasps. On the basis of a well-resolved phylogeny, we reconstruct the evolutionary history of antennal coiling and associated morphological modifications to study the mode of evolution of this complex character system. Results Our study reveals a large variation in shape, location and ultra-structure of male-specific modifications on the antennae. As for antennal coiling, we find either single-coiling, double-coiling or the absence of coiling; each state is present in multiple genera. Using a model comparison approach, we show that the possession of antennal modifications is highly correlated with antennal coiling behaviour. Ancestral state reconstruction shows that both antennal modifications and antennal coiling are highly congruent with the molecular phylogeny, implying low levels of homoplasy and a comparatively low speed of evolution. Antennal coiling is lost on two independent occasions, and never reacquired. A zero rate of regaining antennal coiling is supported by maximum parsimony, maximum likelihood and Bayesian approaches. Conclusions Our study provides the first comparative evidence for a tight correlation between male-specific antennal modifications and the use of the antennae during courtship. Antennal coiling in Diplazontinae evolved at a comparatively low rate, and was never reacquired in any of the studied taxa. This suggests that the loss of antennal coiling is irreversible on the timescale examined here, and therefore that evolutionary constraints have greatly influenced the evolution of antennal courtship in this group of parasitoid wasps. Further studies are needed to ascertain whether the loss of antennal coiling is irreversible on larger timescales, and whether evolutionary constraints have influenced courtship behavioural traits in a similar way in other groups.

Factor Structure of the Neck Disability Index

BACKGROUND CONTEXT: The Neck Disability Index frequently is used to measure outcomes of the neck. The statistical rigor of the Neck Disability Index has been assessed with conflicting outcomes. To date, Confirmatory Factor Analysis of the Neck Disability Index has not been reported for a suitably large population study. Because the Neck Disability Index is not a condition-specific measure of neck function, initial Confirmatory Factor Analysis should consider problematic neck patients as a homogenous group. PURPOSE: We sought to analyze the factor structure of the Neck Disability Index through Confirmatory Factor Analysis in a symptomatic, homogeneous, neck population, with respect to pooled populations and gender subgroups. STUDY DESIGN: This was a secondary analysis of pooled data. PATIENT SAMPLE: A total of 1,278 symptomatic neck patients (67.5% female, median age 41 years), 803 nonspecific and 475 with whiplash-associated disorder. OUTCOME MEASURES: The Neck Disability Index was used to measure outcomes. METHODS: We analyzed pooled baseline data from six independent studies of patients with neck problems who completed Neck Disability Index questionnaires at baseline. The Confirmatory Factor Analysis was considered in three scenarios: the full sample and separate sexes. Models were compared empirically for best fit. RESULTS: Two-factor models have good psychometric properties across both the pooled and sex subgroups. However, according to these analyses, the one-factor solution is preferable from both a statistical perspective and parsimony. The two-factor model was close to significant for the male subgroup (p<.07) where questions separated into constructs of mental function (pain, reading headaches and concentration) and physical function (personal care, lifting, work, driving, sleep, and recreation). CONCLUSIONS: The Neck Disability Index demonstrated a one-factor structure when analyzed by Confirmatory Factor Analysis in a pooled, homogenous sample of neck problem patients. However, a two-factor model did approach significance for male subjects where questions separated into constructs of mental and physical function. Further investigations in different conditions, subgroup and sex-specific populations are warranted.

Impact of antibiotic use on carbapenem resistance in Pseudomonas aeruginosa: is there a role for antibiotic diversity?

In this study, we aimed to evaluate the relationship between the rates of resistance of Pseudomonas aeruginosa to carbapenems and the levels and diversity of antibiotic consumption. Data were retrospectively collected from 20 acute care hospitals across 3 regions of Switzerland between 2006 and 2010. The main outcome of the present study was the rate of resistance to carbapenems among P. aeruginosa. Putative predictors included the total antibiotic consumption and carbapenem consumption in defined daily doses per 100 bed days, the proportion of very broad-spectrum antibiotics used, and the Peterson index. The present study confirmed a correlation between carbapenem use and carbapenem resistance rates at the hospital and regional levels. The impact of diversifying the range of antibiotics used against P. aeruginosa resistance was suggested by (i) a positive correlation in multivariate analysis between the above-mentioned resistance and the proportion of consumed antibiotics having a very broad spectrum of activity (coefficient = 1.77; 95% confidence interval, 0.58 to 2.96; P < 0.01) and (ii) a negative correlation between the resistance and diversity of antibiotic use as measured by the Peterson homogeneity index (coefficient = -0.52; P < 0.05). We conclude that promoting heterogeneity plus parsimony in the use of antibiotics appears to be a valuable strategy for minimizing the spread of carbapenem resistance in P. aeruginosa in hospitals.

Rapid screening for carriage of methicillin-resistant Staphylococcus aureus by PCR and associated costs

PCR tests for the rapid and valid detection of methicillin-resistant Staphylococcus aureus (MRSA) are now available. We evaluated the costs associated with contact screening for MRSA carriage in a tertiary-care hospital with low MRSA endemicity. Between 1 October 2005 and 28 February 2006, 232 patients were screened during 258 screening episodes (644 samples) for MRSA carriage by GenoType MRSA Direct (Hain Lifescience GmbH, Nehren, Germany). Conventional culture confirmed all PCR results. According to in-house algorithms, 34 of 258 screening episodes (14.7%) would have qualified for preemptive contact isolation, but such isolation was not done upon negative PCR results. MRSA carriage was detected in 4 (1.5%) of 258 screening episodes (i.e., in four patients), of which none qualified for preemptive contact isolation. The use of PCR for all 258 screening episodes added costs (in Swiss francs [CHF]) of CHF 104,328.00 and saved CHF 38,528.00 (for preemptive isolation). The restriction of PCR screening to the 34 episodes that qualified for preemptive contact isolation and screening all others by culture would have lowered costs for PCR to only CHF 11,988.00, a savings of CHF 38,528.00. Therefore, PCR tests are valuable for the rapid detection of MRSA carriers, but high costs require the careful evaluation of their use. In patient populations with low MRSA endemicity, the broad use of PCR probably is not cost-effective.

Decreasing risk of hepatitis a infection in león, nicaragua: evidence from cross-sectional and longitudinal seroepidemiology studies

BACKGROUND AND OBJECTIVES Nicaragua is highly endemic for hepatitis A. We aimed to provide an estimate of the change in the age-specific risk of hepatitis A virus (HAV) infection based on serological data from cross-sectional and longitudinal samples collected in León, Nicaragua, in 1995/96 (n = 979) and 2003 (n = 494). METHODS The observed age-specific prevalence of anti-HAV antibodies was correlated to the age-specific risk of infection by calculating the probability of freedom from infection at a specific age. RESULTS The proportion of seropositive children aged 1.5 to 6 years was 42% in 2003 compared to 67% in 1995/96. Estimated annual risk of infection for a 3-year old child was 30% (95% CI: 27.0%, 33.1%) in 1995 and 15.5% (95% CI: 12.4%, 19.0%) in 2003. There was good agreement between estimates based on cross-sectional and longitudinal data. The age-specific geometric mean of the quantified anti-HAV antibody levels assessed in 2003 was highest at age 4 and decreased steadily up to age 40. CONCLUSIONS The substantially lower risk of HAV infection in 2003 than in 1995 for young children indicates a beginning transition from high to intermediate endemicity in León, Nicaragua. Consecutive age-stratified serosurveys are useful to assess changes in risk of infection following public health interventions. The decreasing age-specific GMC of anti-HAV antibodies during adulthood in a country with endemic HAV indirectly suggests that ongoing HAV exposure in the community has marginal boosting effect on antibody levels once protective immunity has been established by natural infection.

THEORETICAL STUDIES ON THE METHODS OF RECONSTRUCTING PHYLOGENETIC TREES FROM DNA SEQUENCE DATA (MOLECULAR EVOLUTION, HOMINOID EVOLUTION, COMPUTER SIMULATION)

(1) A mathematical theory for computing the probabilities of various nucleotide configurations is developed, and the probability of obtaining the correct phylogenetic tree (model tree) from sequence data is evaluated for six phylogenetic tree-making methods (UPGMA, distance Wagner method, transformed distance method, Fitch-Margoliash's method, maximum parsimony method, and compatibility method). The number of nucleotides (m*) necessary to obtain the correct tree with a probability of 95% is estimated with special reference to the human, chimpanzee, and gorilla divergence. m* is at least 4,200, but the availability of outgroup species greatly reduces m* for all methods except UPGMA. m* increases if transitions occur more frequently than transversions as in the case of mitochondrial DNA. (2) A new tree-making method called the neighbor-joining method is proposed. This method is applicable either for distance data or character state data. Computer simulation has shown that the neighbor-joining method is generally better than UPGMA, Farris' method, Li's method, and modified Farris method on recovering the true topology when distance data are used. A related method, the simultaneous partitioning method, is also discussed. (3) The maximum likelihood (ML) method for phylogeny reconstruction under the assumption of both constant and varying evolutionary rates is studied, and a new algorithm for obtaining the ML tree is presented. This method gives a tree similar to that obtained by UPGMA when constant evolutionary rate is assumed, whereas it gives a tree similar to that obtained by the maximum parsimony tree and the neighbor-joining method when varying evolutionary rate is assumed. ^

Phylogenetic Relationships of Clawed Lobster Genera (Decapoda : Nephropidae) Based on Mitochondrial 16S rRNA Gene Sequences

Approximately 350 base pairs (bp) of the mitochondrial 16S rRNA gene were used to study the phylogenetic relationships among 5 genera of the clawed lobster family Nephropidae (infraorder Astacidea), including Homarus, Homarinus, Metanephrops, Nephrops, and Nephropsis. Maximum-parsimony analysis, using a hermit crab, Pagurus pollicaris (infraorder Anomura), as an outgroup. produced a tree topology in which Homarus and Nephrops formed a well-supported clade that excluded Homarinus. The same tree topology was obtained from both neighbor-joining and maximum-likelihood analyses, Some morphological characters that appear synapomorphic for Nephrops and Metanephrops may be due to convergence rather than symplesiomorphy. The current taxonomy, therefore, does not reflect the phylogeny of this group as suggested by the molecular data. More molecular data and studies using homologous morphological characters me needed to reach a better understanding of the phylogenetic history of clawed lobsters.

Comparative assessment of ELISAs using recombinant saposin-like protein 2 and recombinant cathepsin L-1 from Fasciola hepatica for the serodiagnosis of human Fasciolosis.

Two recombinant Fasciola hepatica antigens, saposin-like protein-2 (recSAP2) and cathepsin L-1 (recCL1), were assessed individually and in combination in enzyme-linked immunosorbent assays (ELISA) for the specific serodiagnosis of human fasciolosis in areas of low endemicity as encountered in Central Europe. Antibody detection was conducted using ProteinA/ProteinG (PAG) conjugated to alkaline phosphatase. Test characteristics as well as agreement with results from an ELISA using excretory-secretory products (FhES) from adult stage liver flukes was assessed by receiver operator characteristic (ROC) analysis, specificity, sensitivity, Youdens J and overall accuracy. Cross-reactivity was assessed using three different groups of serum samples from healthy individuals (n=20), patients with other parasitic infections (n=87) and patients with malignancies (n=121). The best combined diagnostic results for recombinant antigens were obtained using the recSAP2-ELISA (87% sensitivity, 99% specificity and 97% overall accuracy) employing the threshold (cut-off) to discriminate between positive and negative reactions that maximized Youdens J. The findings showed that recSAP2-ELISA can be used for the routine serodiagnosis of chronic fasciolosis in clinical laboratories; the use of the PAG-conjugate offers the opportunity to employ, for example, rabbit hyperimmune serum for the standardization of positive controls.

Q fever outbreak in the terraced vineyards of Lavaux, Switzerland

Coxiella burnetii infection (Q fever) is a widespread zoonosis with low endemicity in Switzerland, therefore no mandatory public report was required. A cluster of initially ten human cases of acute Q fever infections characterized by prolonged fever, asthenia and mild hepatitis occurred in 2012 in the terraced vineyard of Lavaux. Epidemiological investigations based on patients' interviews and veterinary investigations included environmental sampling as well as Coxiella-specific serological assay and molecular examinations (real-time PCR in vaginal secretions) of suspected sheep. These investigations demonstrated that 43% of sheep carried the bacteria whereas 30% exhibited anti-Coxiella antibodies. Mitigation measures, including limiting human contacts with the flock, hygiene measures, flock vaccination and a public official alert, have permitted the detection of four additional human cases and the avoidance of a much larger outbreak. Since November 2012, mandatory reporting of Q fever to Swiss public health authorities has been reintroduced. A close follow up of human cases will be necessary to identify chronic Q fever.