Anopheles gambiae s.l. with separate parametrization schemes for Anopheles gambiae s.s. and Anopheles arabiensis. The characterizations were constructed based on literature from the past decades. This part of the study is focusing on the model’s ability to separate the mean state of the two species of the An. gambiae complex in Africa. The model is also evaluated with respect to capturing the temporal variability of An. arabiensis in Ethiopia. Before conclusions and guidance based on models can be made, models need to be validated.The first part of this study aimed to develop a model for An. gambiae s.s. and An. arabiensis was constructed. An additional 992 points were collected on the presence An. gambiae s.l.. These data were used to assess if the model could recreate the spatial distribution of the two species. The dataset is made available in the public domain. This is followed by a case study from Madagascar where the model’s ability to recreate the relative fraction of each species is investigated. In the last section the model’s ability to reproduce the temporal variability of An. arabiensis in Ethiopia is tested. The model was compared with data from four papers, and one field survey covering two years.The model used in this paper is described in part one . For the validation of the model, a data base of 5,935 points on the presence of An. gambiae s.s. and An. arabiensis in sub-Saharan Africa. This implies this model can be used for seasonal and long term predictions of changes in the burden of malaria. Before models can be used to improving human health, or guide which interventions are to be applied where, there is a need to understand the system of interest. Validation is an important part of this process. It is also found that one of the main mechanisms separating An. gambiae s.s. and An. arabiensis is the availability of hosts; humans and cattle. Climate play a secondary, but still important, role.Overall, the model has a realistic representation of seasonal and year to year variability in mosquito densities in Ethiopia. The model is also able to describe the distribution of  Anopheles gambiae s.s. and Anopheles arabiensis[Several attempts have been made to map the distribution of rabiensis-5, two orabiensis showed trabiensis and inserabiensis. Often, rabiensis.Anopheles arabiensis inhabits areas from South Africa in the south to Mauritania and Sudan in the north. In Central-West Africa there is a pocket with very few observations of An. arabiensis. The border of this pocket is formed by Angola, Zambia, Burundi, Rwanda, Uganda, South-Sudan, Central African Republic, Congo, Gabon, and Equatorial Guinea. Anopheles gambiae s.s. is currently separated into five chromosomal forms: Forest, Bamako, Savanna, Mopti and Bissau [d Bissau , and twod Bissau ,11. It id Bissau .An. gambiae s.s. and An. arabiensis[Recent studies have shown that interventions aimed to prevent malaria has an impact on balance between rabiensis. The relrabiensis. In Tanzrabiensis. Given tAn. gambiae s.s. and An. arabiensis The model, which is described in part one [This paper is the second of two describing and validating a new model of the dynamics of part one , is a bipart one .In addition to predicting the current distribution, these type of models can be used to project changes in the historical and future density and distribution of these species. They can describe changes from day-to-day, month-to-month, year-to-year, and decade-to-decade. The model, named Open Malaria Warning (OMaWa) , includeAn. gambiae s.s. and An. arabiensis, with simulations covering an entire continent. It is also the first time age dependent life history and mosquito dispersion (spread of mosquitoes) has been included in a continental analysis. The model is validated against 6,927 presence/absence points of the two species, and a more detailed analysis is carried out for Madagascar. The data is freely available to the public [This is the first time a biophysical model has been used to model the relative density of e public . This stAn. arabiensis and An. gambiae s.s.[et al[An. arabiensis, 1,813 for An. gambiae s.s., and 992 for An. gambiae. Merging these data with the three databases [An. arabiensis, 3,009 for An. gambiae s.s., and 992 for An. gambiae[To date there are three data sets describing the occurrence of biae s.s.,19,20. A.s.[et al. To comp. gambiae. Three m. gambiae, and occAn. gambiae was mainly used to estimate the occurrence of An. gambiae s.l. in Namibia, DRC, South Sudan, Angola, Congo, and northern South Africa. To classify the points the expert opinion polygons from Sinka et al[An. arabiensis polygon only was classified as An. arabiensis, points falling within the An. gambiae s.s. polygon only as An. gambiae s.s., and points falling within both polygons were assigned both species. To classify true presence/absence points the data described previously was used. Observations of An. gambiae s.s. were classified as presence for this species. Absence points for An. gambiae s.s. were those where An. arabiensis had been recorded, and no An. gambiae s.s. had been observed within a radius of 100 km. The same approach was used for An. arabiensis.The database containing nka et al was usedet al[et al[An. arabiensis and An. gambiae s.s.. To reproduce the images in the paper the figures were geo-referenced, and polygons were drawn based on the 0.65-1 probability. The selection was based on the colouring they used in the figure. Next a 50 by 50 km grid was overlaid with the polygons, and points falling within the polygons were classified as presence points. Points falling outside were classified as absence. The second paper is by Levine et al[et al[et al[R + 0.587 · G + 0.114 · B)/255. This new raster image was then gridded to a 50 by 50 km grid. Presence was defined as probability greater than approximately 0.4. As for Rogers et al[et al[et al[This model (OMaWa) was compared with species predictions from four other models, as well as the expert opinion from Sinka et al. The fir al[et al where thine et al. They us al[et al. Since t al[et al was used al[et al.An. gambiae s.s. and An. arabiensis data from Pock Tsy’s et al[To investigate if the model is able to estimate the relative fraction of y’s et al and Chauy’s et al article a) For each observation there are information about the month of collection as well as longitude and latitude. From the model data, covering the period 1990-2008, the closest point to each observation in the month of collection is selected, and the yearly monthly mean is calculated. These data were used to make box plots, weighting for the number of observations in each point, comparing the observations with the model. b) From the data produced in a, maps were created using a distance weighted kernel with cut off at 100 km. Hence observations further away than 100 km were not included, and closer points will be given more weight. c) The distance to the closest wrong (difference in fraction greater than 0.2) and correct  prediction will be indexes for the spatial accuracy. A non-parametric test like the Wilcoxon rank sum test with continuity correction (Mann-Whitney) test can then be used to test if the two indexes differ by a location shift of zero, and the alternative is that they differ by some other location shift.An. gambiae s.s. was to test if the model had a general validity, not limited to Ethiopia. Two high resolution runs only covering Ethiopia were done; one at 30 km, covering the period from 2000 to 2006 (Eth30), and one at 18 km (Eth18) covering the period from 2008 to 2011. These two runs differ from the one covering all of Africa in the way that the weather simulations were forced to follow the observed weather pattern. The technique used to accomplish this is called spectral nudging. In the African run (TC50) the intention was not to reproduce the exact year to year variability, but the interest was to reproduce reasonable weather in a reasonable climate, and thus no nudging was used. To validate the ability to reproduce seasonal variations data from Eth30 and Eth18 to drive OMaWa was used.In addition to looking at the spatial patterns, it is of interest how the model reproduce temporal variability in mosquito numbers. Originally, this model was developed to increase the understanding of malaria epidemiology in Ethiopia. The motivation of introducing For simulations driven by Eth30 the model was run without dispersion, BLL aquatic mortality, development rate with no species correction, default gonotrophic cycle, and AL adult mortality. TC50 and Eth18 were run with the following parametrization: with dispersion, KBLL aquatic mortality, development rate with species correction, default gonotrophic cycle, and BLLad adult mortality. All results are based on single realizations of the model, and error bars are therefore not reported.There are few papers describing the year to year, and seasonal variations in mosquito numbers in Ethiopia. In the validation process three papers were used, one master thesis, and field data from Chano Mille, Arba Minch describing mosquito seasonality.et al[An. arabiensis larva density in the vicinity of six villages in central Ethiopia, December 2007 to June 2008. The second paper is by Taye et al[An. arabiensis numbers in Sille (Southern Ethiopia). The third paper is by Yemane Ye-Ebiyo et al[The first, a paper by Kenea et al, is descaye et al and is riyo et al, where tAn. arabiensis in Awash Valley, Ethiopia, was described [To compare the absolute density, it would be required that the papers reported the larva/mosquito density per square kilometre over a larger area. Since this is not the case, scaling is necessary. The last study is by Balkew, where the seasonality of escribed . The stuIn addition to the published data, Fekadu Massebo collected one year (May 2009 to April 2010) of mosquito densities in Chano Mille, Ethiopia. The study site is described in ,31. To sAll correlations (Pearson) are calculated from the values reported in the papers -29, and S and 37°N, with 27 vertical levels, ranging from the surface to pressure p = 10 hPa. Inside the channel, a domain with 30 km resolution was set up. This domain has boundaries at 25.56°E, 53.18°E, 0.24°N, and 19.29°N. To ensure the model reproduced the observed year to year anomalies, the model was nudged, using spectral nudging, against waves  longer than 1,000 km in both domains. The Kain Frisch cumulus parametrization scheme was used [The simulations in this paper was driven by three different realizations of a limited area climate model. The first realization (Eth30), carried out in 2009, comes from WRF model version 3.1.1 . It was was used ,34.The second realization (TC50), carried out in 2011, had again a tropical channel set up. The model was run at 50 km resolution from January 1 1989 to January 1 2009. At the north and southern boundaries the model was driven by Era Interim. The Kain Frisch cumulus parametrization scheme was used ,34. No nN, 50.99°N, 1.45°S, and 18.97°E.In the third experiment (Eth18), done in 2012, WRF 3.3.1 was used with the Tiedtke cumulus parametrization scheme ,36. The The Regional Committee for Medical and Health Research Ethics, Western Norway, and the Ethical Committee of the Faculty of Medicine of Addis Ababa University and The National Health Research Ethics Review Committee (NERC) of Ethiopia granted ethical approval for the study.An. gambiae was focused on areas where little information about the occurrence of An. arabiensis and An. gambiae s.s. was available. Figure An. arabiensis and An. gambiae s.s.. The white contours are indicating the presence of each species. The pattern is consistent with the general perception of the species range [An. gambiae s.s. in Ethiopia. Still there are some unresolved issues. To date there are no records of An. arabiensis in Côte d’Ivoire; no models, this included, have been able to model the absence of this species in Côte d’Ivoire. A look at the figure also reveals some probable inconsistencies with respect to the species distribution in southern Chad where An. arabiensis should be dominating [An. quadriannulatus. There are however no recent available surveys of An. gambiae s.l. in the states of Gauteng, North West or South Western Limpopo. In Namibia, where An. gambiae s.l. has been observed as far south as -23.7°N[N. Since there are no available data on the recent distribution of this complex in Namibia, it is difficult to know whether the model is correct or wrong. The model also suggests An. gambiae is absent in large parts of Gabon. Previous studies have found An. gambiae in Lambarene [et al later found An. gambiae in Port-Gentil [et al[An. gambiae s.s. in Haut-Ogooué, which was also predicted by the model. In the north-eastern part of Gabon it has not been possible to find any recent mosquito surveys, and it is therefore hard to conclude if the predicted absence of An. gambiae in this region is correct.Figure es range . This ises range -4 has beminating . In Soutminating , althougs -23.7°N, the modambarene  and Moyeambarene , while Mambarene . It shout-Gentil , as predt-Gentil  did not il [et al also fouet al[et al[An. arabiensis is wrong over West Africa and Southern Cameroon. A mosquito survey in Namibia, and north-eastern Gabon, would also clarify the present-day species composition in these countries.To evaluate the quality of the model with respect to classifying the presence and absence of the species the methodology described previously was used. Table et al Sinka et al[et al are greaAn. arabiensis and An. gambiae s.s., the island is well suited to address whether the model is able to reproduce the relative fraction of each species.Three measures to evaluate the model was defined. For method a) the mean absolute error was 0.22. The box plot in Figure An. arabiensis from the model, grouped by the fraction in the observations. It is clear, while capturing the main tendencies well, the model has problems with the exact separation between the two species. In the mixed group, the model tends to let one species dominate over the other, possibly letting An. arabiensis dominate too easily.Since Madagascar has a sharp separation between An. arabiensis as modelled, and observed. An eyeball comparison shows the separation is shifted westward in the model, and a bias in the South-Eastern tip of Madagascar. Whether this is a result of (climate) model resolution, failing to accurately separating the west/east gradient in topography, or the biological parametrization being inaccurate is hard to quantify. It is hoped this can be tested in a future analysis with higher model resolution.Figure p < .0001). Thus, although with biases, it is concluded that distance to closest correct prediction and closest wrong prediction are non-identical populations.Table An. arabiensis and An. gambiae s.s.. These results are meant to complement the continental analysis. Eth30 and Eth18 refers to the weather data used to drive OMaWa.It is important that mosquito models reproduce the seasonal cycle correctly, since this will be an indication of the sensitivity to climate. Here results from the model are compared to a number of observational studies. The comparison with each individual study might not have much information, but it is recommended that readers look at the results as a whole, having in mind the continental analysis showing the model is able to separate the distribution of et al reported the An. arabiensis larva density in six locations in central Ethiopia, December 2007 to June 2008. Five of the sites followed the same seasonality, while one had the highest density before the rainy season started. The model is not designed to capture such local variations, but is rather aiming to describe the median, or sometimes mean, state within a certain area. In their study all anopheline positive habitats present within a 500 m radius of each irrigated village/town and 700m along the major drainages (lake or river) were sampled. This means that the data should be comparable to what is modelled. The seasonality of larva density, Ap, is then calculated as Cl is a dimensionless constant. Correlations with the median relative seasonality, model vs. Kenea et al., is 0.97, and mean relative seasonality 0.92. The observations and modelled results can be seen in Figure In this study  Kenea etet al[An. arabiensis in Sille, Ethiopia. For simplicity it is assumed the human biting rate is independent of temperature and availability of breeding sites. This means the relative monthly mean sum of mosquitoes from the model should be directly comparable with the records from the paper. The model seems to under-predict the relative abundance of An. arabiensis in October 2001, and over-predict the rise in mosquito numbers in February. Otherwise the modelled number of mosquitoes seems comparable to what was observed by Taye et al. The correlation between observations and model (2001-2002) is 0.91. The observations and model results are shown in Figure In 2001-2002 Aseged Taye et al recordedC is a constant, and that the samples are representative for a larger area, the relative number of larva in that area can be estimated as LPD · Wa, where Wa is the mean water area in m2. This way it is assumed the number of puddles is constant from July to September, and that the puddles only change their surface area. These values are roughly comparable to the modelled number of larva. Since only the latitude (and not the longitude) is reported in the paper, and Zwai is not located at latitude 9°N, model data between longitudes 38.69 to 39.23°E and latitudes 7.88 to 8.42°N, an area covering Zwai, were selected. Using this method correlation is 0.99(0.321.00). Confidence interval is estimated using 1,000 random samples of the points within the bounding box, and the 2.5% and 97.5% quantiles of the correlations is reported. Since the sample size is small and the data might not be directly comparable, the correlation should be interpreted with care. The data from the observations and the model can be seen in Figure If it is assumed larva per dip has units E and latitudes 8.75 to 8.92°N. The data are based indoor space spray collections. Since the malaria model was not run for 1999, and 2000 is considered as a spin-up year, the multi-year monthly mean for the years 2001-2006, and 2008-2009 was used . The observations are compared to the scaled sum of mosquitoes of all age groups, which should be comparable to what was reported in the thesis. Correlations in Buse + Gelcha (two locations described in the thesis) was 0.75, 0.79 for Sugar Estate, and 0.76 for Metehara Town. Confidence intervals are not reported for Metehara Town since the number of observations are low. The data can be seen in Figure This study was carried out in 1999-2000 in Metehara at longitudes 39.50 to 40.00°An. arabiensis was very low (Figure As seen in Figure w Figure . It is iEach of the five case studies consist of short time series, with different observational methodologies. It was attempted to show how the model results can be compared to the different type of observations, and in general the model is in good agreement with the observations. Since none of the studies cover several years, it was only possible to validate whether the model captured the seasonal cycle in mosquito numbers. The good agreement with all of the five case studies, means the model probably responds correctly to the environment, and thus it is likely OMaWa can reproduce year-to-year variability as well.An. gamibiae s.s. and An. arabiensis across Africa, and that it is able to model the seasonal and year-to-year variations in mosquito densities. While the results are robust with respect to the mean distribution and density, there is a sampling bias related to the recent distribution in DR Congo, northern South Africa, southern Namibia, eastern Angola, Central African Republic, eastern Gabon, eastern Chad, South Sudan, and Somalia. This implies models can not be robustly validated in these regions, and that long term changes in the species composition can not be addressed. For the temporal variability, the model has only been validated for Ethiopia, using short time series. Although the model matches well with the observations, most of the time series are short, implying the ability to reproduce year-to-year variations has not been fully addressed.In this paper, the model has been validated using independent data. The model was designed to have a general validity, not being restricted to a specific locality. The study shows the model can capture the distribution and density of An. arabiensis. Similarly, the presence of An. gambiae s.s. is linked to the presence of humans, modulated by the density of An. arabiensis. Water and air temperature, and availability of breeding sites play secondary roles for the continental distribution of these species, but might be locally important in margin zones. The recent distribution shifts in species composition observed in Kenya [An. gambiae s.s. due to interventions like IRS and LLINs. An alternative explanation might be the competitive advantage of An. arabiensis efficiently feeding on cattle, and thus suppressing the number of An. gambiae s.s. through easier access to blood, and thus reproducing at a higher rate. Over time, these interventions mainly reduce the human biting rate, and not necessarily the longevity of mosquitoes; the most efficient measure in MacDonald’s formula of the basic reproductive number. Next, it can be challenged if a reduction of the number of breeding sites, lowering the number of adult mosquitoes per human, would be as efficient, and cost-effective, as IRS and LLINs over time. Studies on the long-term effect of interventions on the mortality rate of mosquitoes is needed to evaluate how these interventions work in practice. The large scale distribution of An. arabiensis, and its relation to cattle distribution, also rises the question of this species is using the odour of bovine to navigate, and if this causes of the observed coexistence of An. arabiensis and cattle. If this applies on large scales, there are reasons to believe the same mechanisms manifest themselves on small scales. In that case, keeping cattle separate from humans should further reduce the human biting rate in areas where An. arabiensis is the dominant species.The results suggest sufficiently high bovine density influences the large-scale distribution of in Kenya ,46 mightAn. gambiae s.s. and An. arabiensis, and how the species have spread, and is evolving, across Africa. In the model, which gives a good representation of the distribution of the two species, An. gambiae s.s. is spreading most efficient on surfaces with continuous human populations, while An. arabiensis disperse more easily on surfaces with continuous cattle populations. It is hypothesized the lack of such a human surface between Kenya and Ethiopia can explain the absence of An. gambiae s.s. in Ethiopia; - to spread to Ethiopia, there is a need of a more or less continuous human population cover from Lake Victoria to southern Ethiopia, sufficient breeding sites, and temperatures which are not too extreme. Thus, not only climate control the presence and absence of these species, but also the availability of hosts. This has implications for the ability to project the future distribution of the two species.Several studies have found out the gene flow of Before models can be related to improving human health, or guide which interventions are to be applied where, there is a need to understand the system of interest. Validation is an important part of this process. Concluding based on too little data, and basing projections of for example the effects of climate change on models which have not been validated, is dangerous , might mThe authors declare that they have no competing interests.An. gambiae s.l.. All authors have contributed to, seen and approved the manuscript.The work presented here was carried out in collaboration between all authors. BL and AS defined the research theme. TML designed methods and mosquito experiments, did the model runs, analysed the data, interpreted the results and wrote the paper. DK, AS and TML designed the regional climate simulations, and evaluated those. FM, MB and TGM collected data for validation, and contributed with comments on the biology of


Gaucher disease (GD) is the most frequent lysosomal storage disorder; type 1 is by far the most common form. It is characterized by variability in age of onset, clinical signs and progression. It is usually diagnosed in the first or second decade of life with the appearance of bone pains, splenomegaly and thrombocytopenia, but the disease may be diagnosed at any age between 1 and 73 years. In the present study, we report 3 cases with late onset of GD in whom the disease was a surprise finding including one patient with Parkinson disease. This late onset is described as an adult form of Gaucher disease.Molecular investigation showed mutational homogeneity in Tunisian adult patients suffering from GD. Indeed, all patients carry the p.N370S mutation: two patients at a homozygous state and one patient at compound heterozygous state.The p.N370S mutation presents a large variability in the onset of the disease and its clinical manifestation supporting the view that GD should be considered as a continuum phenotype rather than a predefined classification. GBA) on1q21 leading to enzymatic deficiency of the β-glucocerebrosidase and the accumulation of glycosylceramide substrate in the macrophage's lysosomes. GD is characterized by a considerable phenotypic and genotypic heterogeneity [GBA gene http://www.hgmd.org. In previous studies, investigation of GD showed that p.N370S, p.L444P and RecNciI mutations were relatively frequent in Tunisian patients [NciI mutations in GBA gene.Gaucher disease (GD) is the most frequent lysosomal storage disorder . It is aogeneity . More thpatients . In the Three patients diagnosed with GD from three unrelated families from southern Tunisia were investigated. The diagnosis was based on the occurrence of hepatosplenomegaly associated with hematological abnormalities and/or bone lesions and was confirmed in all patients by the presence of Gaucher cells.XhoI and NciI to screen for p.N370S and p.L444P respectively. Sequencing was performed on an ABI 3130 to detect the three mutations .After the patient's written informed consent, 5 ml of blood were collected. DNA was extracted from peripheral blood mononuclear cells using the standard salting-out procedure. DNA samples were then amplified by polymerase-chain-reaction (PCR) followed by enzymatic restriction using 3). Clinical examination showed a voluminous splenomegaly associated with pancytopenia. Neurological symptoms were absent. The diagnosis of GD was confirmed by bone marrow examination . Family history showed no particular pathologies. The diagnosis of GD was confirmed by a myelogram and bone marrow biopsy that revealed the presence of Gaucher cells  and splenomegaly. The diagnosis of GD was confirmed by myelogram  developed Parkinson disease at age 52. The co-morbidity of GD and Parkinson disease has been largely discussed -11. The borators . Whereasborators .NciI) is the best alternative to reduce the incidence of the severe form of the disease.It has been suggested that only one third of the patients who were homozygote for p.N370S mutation come to medical attention and that about two thirds remain asymptomatic throughout life . CarrierThere is a great variability in the clinical manifestation as well as in the onset and course of GD. This supports the need for a new classification and reinforces the hypothesis of the concept of a phenotypic continuum. Clinicians, especially in hematology and internal medicine, should suspect the diagnosis of GD in any adult patient suffering from abdominal and/or bone pain with unexplained hematological disorder especially in combination with organomegaly.GD: Gaucher disease; PCR: polymerase chain reaction; PD: Parkinson disease; RFLP: Restriction fragment length polymorphism.The authors declare that they have no competing interests.SA had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. Study concept and design: SA and SM. Acquisition of data: FBR, FK, RK, WC, MN, HA, IJ and ME. Analysis and interpretation of data: FBR, RK, WC, MN, HA, SA and SM. Drafting the manuscript: FBR. Critical revision of the manuscript for important intellectual content: SA, RK, HA, SM. Obtained funding: SA. Administrative, technical or material support: FBR, RK, WC, MN, and HA. Study supervision: SA, ME and SM. All authors read and approved the final manuscript.


Pedaliodes poesiaChusquea scandens  is the larval food plant. Eggs are laid singly or in pairs on the bottom side of host plant leaves. The duration of the egg, larval, and pupal stages, combined, is 99–107 days.We describe the immature stages  Pedaliodes poesia .s poesia   and northern  Peru on the eastern Andean slope, throughout the eastern slope of Ecuador, in all three Cordilleras of Colombia, including the Pacific slope of the Western Cordillera, and in the El Tamá range of south-western Venezuelan  bands .nezuelan . Similarm) bands . Howeversemblage . In EcuaP. poesia was vaguely defined as “New Granada” , and the presence of red patches on the dorsal forewing (FW) ; to P. suspiro Adams and Bernard in the Sierra de Perijá (Colombia); and in the south to P. hewitsoni Staudinger, found in southeastern Peru  and northern Bolivia. All these taxa are morphologically and ecologically so closely-related, that they may eventually be considered a single species (Pyrcz pers. obs.). Sexual dimorphism in P. poesia is well developed, particularly when compared with other species of Pedaliodes. It is expressed mostly in the much lighter ventral HW pattern of the females, marked with wide, whitish or silvery patches and bands. Females are also slightly larger, and have somewhat more undulated outer margins of the HW. In some populations, females bear an orange or reddish dorsal FW patch, but this is never apparent in males. Sexual dimorphism in P. poesia and related species is so pronounced that it has previously led to the description of the two sexes as separate species. For example, Pronophila phanaraea, described by Hewitson (1868) is in fact a female of P. poesia from Ecuador  . Some of Ecuador . Similarewitsoni . Within Pedaliodes use bamboo species in the genus Chusquea (Poaceae) as their primary host plant . Despite the fact that over 270 species of Pedaliodes are distributed throughout the Andes (P.phoenissa (Hewitson)   and P. z Heredia . Here weWe carried out rearing and field investigations at the Yanayacu Biological Station and Center for Creative Studies , located in Napo Province, in the Andes of northeastern Ecuador. The study site is located approximately five kilometers west of the town of the town of Cosanga, adjacent to Cabañas San Isidro, and includes around 2000 hectares of primary cloud forest bordered by cattle pasture and other disturbed habitats , are siP. poesia and P. zingara is the light brown first instar head capsule of P. poesia, in contrast with the shining black head of first instar P. zingara. Also, while Heredia and Viloria (P. zingara, our observations suggest that P. poesia may take as much as a month longer to develop from egg to adult. Schultze did not specify the complete life-cycle duration of P. phoenissa, but the period from third instar to imago lasts roughly 50 days, which indicates a development time similar to that of P. poesia. Heredia and Viloria (P. zingara (when compared to Parapedaliodes parepa and P. phoenissa; Apart from variation in pupal shape, the most striking difference between  Viloria  report a Viloria  suggest P. zingara and P. poesia compared to those of P.parepa (P. parepa are rounder in cross-section, rather than flattened across the top, and are thus less trapezoidal in cross section than the two Pedaliodes spp. Similarly, the head scoli of Parapedaliodes appear to be somewhat more conical or pointed than the rounded, robust scoli of Pedaliodes. These caterpillars are similar in coloration, but Parapedaliodes exhibits a shining black head capsule like that of P. zingara. The strongly angular shape of Parapedaliodes pupae is similar to P. zingara. Comparing gross morphology of all three, it appears that the pupa of P. zingara is somewhat intermediate between P. poesia and Parapedaliodes in its “angularity.”We found few differences between larvae of P.parepa . It appeP. poesia larvae feed on Chusquea, a widespread bamboo integral to the Andean landscape. Nevertheless, our knowledge of pronophiline natural history is in its infancy. As the morphology and behavior of immature stages are known to provide important phylogenetic information in butterflies (e.g., Like most Pronophilina, es e.g., , we encoPaper copies of this article will be deposited in the following libraries. Senckenberg Library, Frankfurt Germany; National Museum of Natural History, Paris, France; Field Museum of Natural History, Chicago, Illinois USA; the University of Wisconsin, Madison, USA; the University of Arizona, Tucson, Arizona USA; Smithsonian Institution Libraries, Washington D.C. USA; The Linnean Society, London, England.


An. gambiae s.l populations in Mbita District and Suba District.Abundance and species composition of sympatric malaria vector species are the important factors governing transmission intensity. A widespread insecticidal bed net coverage may replace endophagic species with exophagic species. However, unique local environments also influence a vector population. This study examined the impacts of insecticidal bed nets on An. gambiae s.l. larvae were compared between 1997, 2009 and 2010 and between geographical areas. The abundance and species composition of An. gambiae s.l. females resting indoors were compared between 1999, 2008 and 2010 and between geographical areas. Bed net coverage was also examined temporally and spatially, and its relationships with vector abundance and species composition were examined.The species compositions of An. gambiae s.s. larvae was 31.4% in 1997, decreasing to 7.5% in 2008 and 0.3% in 2010. The density of indoor resting An. gambiae s.l. females decreased by nearly 95%, and the relative abundance of An. gambiae s.s. females decreased from 90.6% to 60.7% and 72.4% in 2008 and 2010, respectively. However, the species composition of indoor resting An. gambiae s.l. females changed little in the island villages, and An. gambiae s.s. remained dominant in the western part of the study area. The density of house resting females was negatively associated with the number of bed nets in a retrospective analysis, but the effect of bed nets on species composition was not significant in both retrospective and cross-sectional analyses.The relative abundance of An. gambiae s.l. populations varies spatially, and locally unique environments are likely to influence the species composition.An increase in bed net coverage does not necessarily replace endophilic species with exophilic species. The effect of bed nets on  Anopheles gambiae s.s. Giles and An. arabiensis Patton, which belong to the An. gambiae complex group (hereafter An. gambiae s.l.). Both species inhabit small sun-lit pools [An. gambiae s.s. is more anthropophagic and endophilic than An. arabiensis[An. gambiae s.s. is considered the most efficient malaria vector in Africa; an area with abundant An. gambiae may have a high transmission risk. However, an area with abundant An. arabiensis does not necessarily have a lower transmission risk, as insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS) are less effective against this exophilic species [Understanding the relative abundance of sympatric malaria vector species is important for vector control, because each species has different vectorial capacity, which reflects on local malaria transmission intensity -3. The mit pools ,5, and oit pools ,7. Howevrabiensis-10. Thus species .An. gambiae s.s. fell compared with An. arabiensis following the introduction of ITNs in Kenya and Tanzania [An. gambiae s.s. On the other hand, ITNs and IRS may not have reduced the abundance of An. gamibae s.s. as much as recent studies have suggested, as its feeding place may have shifted outdoors to avoid insecticides [Their ecological differences imply that an introduction of ITNs or IRS alters the vector species composition. Recent studies have reported that the relative abundance of indoor resting Tanzania ,12-16. ATanzania . Neverthcticides ,18,19.Anopheles arabiensis often dominates in drier seasons and areas [Moreover, this phenomenon requires further careful interpretation, as an environmental change can also influence local mosquito species composition. nd areas ,20-22, tAn. gambiae s.l. populations in Mbita District and Suba District in the Nyanza Province of western Kenya. In particular, we wanted to confirm if ITNs had reduced densities of the indoor resting malaria vectors and altered their species composition at both adult and larval stages, and to reveal if the effects of ITNs on their density and abundance were geographically heterogeneous.The main objective of this study was to reveal if an introduction of ITNs had altered 2, however, the hilly regions were excluded from this study as malaria vectors are not abundant in these areas. The study area was mostly deforested by the mid-1990s with the exception of a few small patches of forest remaining in the hills, the vegetation cover had remained mostly unchanged since then. The rainfall pattern in the area is bimodal, with a long rainy season occurring from March through May, and a short rainy season occurring around November.Mbita District and Suba District lie within the Kenyan part of the Lake Victoria basin. The study area included five inhabited islands, Kibuogi, Mfangano, Ngodhe, Rusinga and Takawiri. Both districts formed a single district (Suba District) prior to 2011, but the western region was later designated as Mbita District. The total area of both districts combined is approximately 1055 kmMost inhabitants belong to the Luo and Suba ethnic groups whose main socio-economic activities are traditional small-scale fishing and farming. Most houses are constructed of a stick framework, plastered with a mixture of mud and cow dung, commonly covered with a corrugated iron roof or, in few instances, with thatch. Kenya’s Ministry of Health, with assistance from the Global Fund, started a distribution of LLINs in this area in late 2006. However, malaria transmission remains high ,24. TherAn. gambiae s.s., An. arabiensis and An. funestus s.s. Giles. A study in 1997 reported that 65.5% of An. gambiae s.l. larvae sampled in the study area were An. arabiensis, and the species composition was spatially heterogeneous [An. gambiae s.s. dominated indoors [The main malaria vectors in the study area are ogeneous ,24,25. H indoors .In May 2009 (during the rainy season), we collected anopheline larvae in nine areas, Gembe, Luanda, Mbita, Nyandiwa, Ogongo, Owichi, Rusinga, Valley South, and Valley North, where a previous study surveyed in February and March, 1997 . Larvae An. gambiae s.l. under a dissecting microscope using morphological keys [An. gambiae s.l. larva were extracted using the ethanol precipitation method. The DNA samples were identified into An. gambiae s.s. or An. arabiensis using the rDNA-polymerase chain reaction (PCR) method [Collected larvae were identified into cal keys . DNAs of) method . In caseIndoor resting anopheline females were collected from 257 houses in 19 villages during May 2008, and from 470 houses in 31 villages during May 2010  and species composition within An. gambiae s.l. (the relative abundance of An. arabiensis) in the study area. A full model included all relevant covariates, this model was then simplified until the Akaike Information Criterion (AIC) was minimized following the stepwise removal of covariates. The significance (P > 0.05) of each fixed variable remaining in the optimal model was estimated using a log-likelihood ratio test. The Statistical Package R version 3.0 was used for all analyses [Maximum likelihood statistical models were used to reveal factors that affect the density of malaria vector species  (employing the R package ‘lme4’) was used to compare the relative abundance of and 2010 . The yeaand 2010 .An. gambiae s.l., An. gambiae s.s. and An. arabiensis from the 11 villages studied between 1999, 2008 and 2010 using a Poisson GLMM. Year was a fixed factor, while village and house were assigned as random factors. In the analyses, two types of geographical variables were also included to explore spatial heterogeneity. We categorized the villages into “island” and “mainland” for the first geographical variable (Island/mainland). The second geographical variable was constructed by grouping the villages using a cluster analysis based on the geographical distance between each pair of villages (employing R package ‘stats’). In these analyses, we could not include the variables related to bed nets and residents, as they were not available for the 1999 data set. Furthermore, we compared the densities of each taxon between years for each village separately, assigning year as a fixed factor and house as a random factor, which would provide us a picture of how the temporal changes in density varied among villages .For indoor resting females, we compared the average numbers of An. arabiensis between years, incorporating the same fixed and random factors used in the analyses of density. Then, we examined temporal change in species composition within each village separately. When no mosquitoes were amplified by PCR for a single house, the house was excluded from the analysis.For adult species composition, we also used a binomial GLMM to compare the relative abundance of An. gambiae s.l. females with the data set from the larger area including 31 villages in 2010, applying the same statistical procedure mentioned above.Using the adult mosquito data set from the 19 villages studied during 2008 and 2010, we revealed the factors affecting the density of each taxon and species composition. As the bed net coverage peaked in the area during 2008, two years after the mass bed net distribution campaign and then began to decline, we wanted to know if this decline affected vector density and species composition. We were also interested in the spatial variations of vector density and species composition, and their changes between 2008 and 2010. For the analyses, we used a Poisson GLMM for density and a binomial GLMM for species composition including house and village as random factors. Fixed factors assigned were year, the number of bed nets, the number of residents and the number of residents that had slept under nets on the previous night in each house, and the two geographical variables used in the analysis for the 11 villages. Furthermore, we examined the spatial variations of vector density and species composition of This study was approved by the Ethical Committee of Kenya Medical Research Institute (KEMRI) (SSC No. 2131), Kenya. After the study was explained to the household heads in the local language, informed consent was obtained from them prior to sampling mosquitoes from their houses and interviewing them about bed net usage.An. gambiae s.l. larvae were collected in 2009. Of these, 28 (7.5%) were An. gambiae s.s., and 346 (92.5%) were An. arabiensis , area was not significant .Since we could not collect enough larvae in Ruri, Owichi and Rusinga in 2009 and in Ruri and Valley North in 2010, the data from these areas were not included in the analysis of species composition. A total of 374 et al. collected 4099 An. gambiae s.l. females from the 11 surveyed villages  were An. gambiae s.s. In 2008, 556 An. gambiae s.l. females were collected from the same 11 villages; 468 of these were used for PCR identification, of which 440 (94.0%) were successfully amplified, identifying 173 An. arabiensis (39.3%), and 267 An. gambiae s.s. (60.7%). In 2010, 865 An. gambiae s.l. were collected, and 730 individuals were used for species identification. Of these, 188 (27.6%) were An. arabiensis, and 492 (72.4%) were An. gambiae s.s.In 1999, Chen An. arabiensis females selected one fixed variable, year, and two random variables, house and village . The relative abundances in 2008 and 2010 were significantly greater than in 1999, and those in 2010 were significantly lower than in 2008.A cluster analysis based on geographical distance grouped the 11 surveyed villages into two separate regions, the eastern region and the western region Figure . The besAn. arabiensis was compared between the years for each village separately, the relative abundance did not differ significantly in Takawiri and Ragwe . The density of An. gambiae s.l. reduced by 93% in 2008 and 2010 when compared with 1999, and the density of An. gambiae s.s. had reduced by 96% in 2008 and 95% in 2010 when compared with 1999 . The density of An. arabiensis reduced by 59% in 2008 and 70% in 2010 compared with 1999 . The model for the number of bed nets also selected island/mainland; however, the fixed variable was not statistically significant . For the numbers of residents and residents having slept under bed nets, the best models also selected geographical region; however, the fixed variable was not statistically significant in either case . The number of bed nets per house decreased by approximately 48% between 2008 and 2010, the number of residents decreased by approximately 10% in 2010, and the number of residents having slept under bed nets also decreased by approximately 23% in 2010 , island/mainland was not statistically significant . The western region had about 1.5 times the number of residents when compared with the other regions .A cluster analysis grouped the 31 villages into three areas, western region, central region and eastern region Figure B. The beAn. gambiae s.l. adults were collected from the 19 villages in 2008  as An. gambiae s.s. In 2010, a total of 1712 An. gambiae s.l. were collected from the same 19 villages, and 1285 of these were used for PCR identification. PCR successfully amplified 1190 (92.6%) samples, of which 546 (45.9%) were An. arabiensis, and 644 (54.1%) were An. gambiae s.s.A total of 1238 An. arabiensis in the 19 villages selected three fixed variables, geographical region, island/mainland and year, and two random variables, house and village . The relative abundance of An. arabiensis was significantly higher in the island population when compared with the mainland population, and higher in the central region and eastern region when compared with the western region . Increasing bed nets by one decreased the average density of An. gambiae s.l. per house by 10%, and increasing residents by one increased it by 13% , the effects of island/mainland and year were not significant . Increasing bed nets by one decreased the average density of An. gambiae s.s. per house by 8%, and increasing residents by one increased it by 13% . Increasing bed nets by one decreased the average number of An. arabiensis by 13%, and increasing residents by one increased it by 13% . The relative abundance of An. arabiensis was higher in the eastern regions than in the western and central regions , the number of residents was significant . Increasing the number of residents by one increased the density of An. gambiae s.l. by 12%, the densities in the central and eastern regions were 42% greater and 30% lower than in the western region, respectively . Increasing the number of residents by one increased the density of An. gambiae s.s. by 11%, and the densities in the central and eastern regions were 19% and 87% lower than in the western region, respectively , the number of bed nets and region were not significant . Increasing the number of residents by one increased the density of An. arabiensis by 5%, and the densities in the central and eastern regions were 3.3 times greater than in the western region, respectively  suggests that the dominancy of An. gambiae s.s. is related to human populations [The dominancy of rabiensis-10, and ulations . AlthougIn addition to the availability of grazing area, the culture associated with the local ethnic groups may influence the abundance of livestock in the study area, and eventually influence the local vector species composition. The traditional lifestyle of the Luo is closely associated with livestock, while the focus of the Suba, who mainly inhabit the islands and the western region, is cultivating crops rather than pasturing livestock. As small boats are the only available transportation to the islands, the limited accessibility may also keep the abundance of livestock low on the islands.Anopheles gambiae s.s. has a lower desiccation tolerance than An. arabiensis, and An. gambiae s.s. tends to dominate in wetter areas when compared with An. arabiensis[An. gambiae s.s. may have higher humidity. Although a climate data set at a fine spatial scale is not available in the study area, the climate in the western region may be wetter than the eastern region. The residents in the western region often plant maize twice a year, but a two-crop system is not common in the eastern region, which suggests that the amount of rainfall is greater in the western region. The remaining patches of indigenous forests also suggest a wetter climate in the western region, and the forests further retain humidity in the area [Climate is another plausible factor for explaining the spatial variation of species composition. rabiensis,22,38. Tthe area . The lakAn. arabiensis was significantly lower in 2010 when compared with 2008. This was attributed to the significant reduction of An. arabiensis density in 2010, and a slight increase in An. gambiae s.s. density. Although the increase of An. gambiae s.s. was not statistically significant, overall An. gambiae s.l. significantly increased in 2010. The increase of vector population may be due to the decrease of bed nets in 2010 versus 2008. The decrease in bed net numbers reduces the selective pressure against An. gambiae s.s., in particular [An. gambiae s.s. [et al. reported that the An. gambiae s.s. had developed insecticide resistance related with a high frequency of knockdown resistance mutation, while An. arabiensis had a comparatively low frequency [An. arabiensis in 2010, as this species had acquired another type of pyrethorid insecticide resistance related to P450 by 2010 [Another interesting phenomenon observed in this study was that the relative abundance of rticular ,12-16. Tiae s.s. ,30,40-42An. arabiensis may simply be an annual variation. The long rainy season in 2008 was slightly drier than in 2010 due to the occurrence of La Niña Modoki [An. arabiensis in 2008. But the wetter rainy season in 2010 should not decrease the actual abundance of An. arabiensis, as its abundance has always been positively associated with rainfall [An. arabiensis is the development of behavioral resistance to ITNs [An. arabiensis is generally exophagic, the An. arabiensis population in the study area may have further enhanced its exophagic behavior, which may have consequently reduced the number of indoor catches of this species using PSC. The larval species composition changed little between 2009 and 2010, which may support this explanation, although larvae were not sampled in the same year.Otherwise, the decrease of a Modoki ,44, whicrainfall -48. Anot to ITNs ,19. AlthAnopheles gambiae s.s. has been replaced by An. arabiensis as the dominant malaria vector species, and has experienced a reduction in density of nearly 95% in the study area over 10 years. These phenomena are most likely explained by the dramatic increase in ITN coverage [An. gambiae s.s. was still dominant in the western part of the study area. The dominance of An. gambiae s.s. is not simply explained by bed net coverage, but rather by other environmental factors such as livestock availability or climate. This study also observed a slight increase in malaria vectors and an increase in the relative abundance of An. gambiae s.s. in 2010 compared with 2008. The decrease of bed net coverage and development of insecticide resistance may partially explain these changes.coverage . HoweverAn. arabiensis as the primary target in a high bed net coverage area. An adequate vector control tool and duration of intervention should be carefully determined for a targeted area [An. gambiae s.s. density in 2010 is alarming. Periodical monitoring of vector density, species composition and insecticide resistance is necessary to detect an unexpected resurgence in malaria vector population and parasite transmission [As this study demonstrated that an increase in bed net coverage does not necessarily replace endophilic species with exophilic species, verifying the target vector species is important in vector control rather than assuming ted area -52. The smission -55. Propsmission .AIC: Akaike Information Criterion; GLMM: Generalized linear mixed model; GPS: Global positioning system; IRS: Indoor residual spraying; ITN: Insecticide treated bed net; KEMRI: Kenya Medical Research Institute; PCR: rDNA-polymerase chain reaction; PSC: Pyrethrum spray catches.The authors declare that they have no competing interests.KF and NM conceived and designed this study. SN helped in designing and planning the study in Kenya. KF, GD, GS and PA collected the field data, and KF, MM, SW, JL and JK organized and conducted the laboratory work. KF performed the data analyses. KF drafted the first manuscript, and KF and NM finalized the manuscript. All authors have read and approved the final manuscript.An. arabiensis larvae. The larvae were sampled in 1997, 2008 and 2010. The parameters for 2009 and 2010 were estimated based on 1997, and the parameters for area was estimated based on island.Results of the best binomial GLMM for the relative abundance of Click here for fileAn. gambiae s.l. females sampled from each villages in 1999, 2008 and 2010.The numbers of Click here for fileAn. arabiensis females in 11 villages. The mosquitoes were sampled in 1999, 2008 and 2010. The parameters for 2008 and 2010 were estimated based on 1999.Results of the best binomial GLMM for the relative abundance of Click here for fileResults of the best binomial GLMMs for the densities of malaria vectors in 11 villages. The mosquitoes were sampled in 1999, 2008 and 2010. Parameters for 2008 and 2010 were estimated based on 1999, and those for the eastern region was estimated based on the western region.Click here for fileResults of the best GLMMs for bed nets, residents and residents slept under nets. The parameters for mainland were estimated based on island, those for the central and eastern regions were estimated based on the western region, and those for 2010 were estimated based on 2008.Click here for fileAn. arabiensis in 19 villages. The mosquitoes were sampled in 2009 and 2010. The parameters for mainland were estimated based on island, those for the central and eastern regions were estimated based on the western region, and those for 2010 were estimated based on 2008.Results of the best binomial GLMM for the relative abundance of Click here for fileResults of the best Poisson GLMM for the densities of malaria vectors in 19 villages. The mosquitoes were sampled in 2008 and 2010. The parameters for 2008 and 2010 were estimated based on 1999, and those for the eastern region was estimated based on the western region.Click here for fileAn. arabiensis in 31 villages. The mosquitoes were sampled in 2008 and 2010. The mosquitoes were sampled in 2010. The parameters for 2008 and 2010 were estimated based on 1999, and those for the eastern region were estimated based on the western region.Results of the best binomial GLMM for the relative abundance of Click here for fileResults of the best Poisson GLMM for the densities of malaria vectors in 31 villages. The mosquitoes were sampled in 2010. The parameters for the central and eastern regions were estimated based on the western region.Click here for file


To the Editor:Rickettsia felis is a bacterium transmitted by the cat flea , which also acts as a reservoir by means of transovarial transmission , identified by using accepted morphologic criteria, were obtained, with each animal yielding one to five fleas. DNA was extracted from each of the 110 flea pools by using a standard silica cartridge method  using the manufacturer’s instructions for tissue DNA extraction. The presence of rickettsial DNA was determined by using the polymerase chain reaction (PCR) with oligonucleotide primers that target rickettesial ompB . Similarity to published sequences was determined with the BLAST program (available from: URL: http://www.ncbi.nlm.nih.gov) hosted by the National Centre for Biotechnology Information.To determine whether ompB and gltA-targeting PCRs. The ompB and gltA DNA sequences of all PCR products were 100% identical to those published for R. felis, thereby providing evidence for the presence of R. felis in fleas collected from >16% of the animals surveyed. PCR-positive fleas were collected from 4 dogs and 14 cats from Bristol, Hampshire, Dorset, and Northern Ireland. Taking into account the number of fleas in each pool, we estimate that 6% to 12% of the fleas collected were infected with R. felis.Eighteen flea DNA pools were positive for spotted fever group rickettsia. All 18 yielded PCR products with both R. felis, has clear public health implications. The bacterium appears to be widely distributed within the country, infecting a geographically dispersed population of Ct. felis. Up to 12% of Ct. felis may be infected with R. felis, a flea that is by far the most common species of ectoparasite encountered on cats and dogs in the U.K. mainland. Furthermore, Ct. felis often feeds on humans.This study represents the first description of a spotted fever group rickettsia endemic to the United Kingdom. The species detected, R. felis. Laboratory confirmation of infection is not easy, but in vitro culture of R. felis, and hence material for a serologic assay for the diagnosis of human R. felis infections, has recently been described, and serology appears to be an accurate indicator of exposure , gentamicin, amoxicillin, and trimethoprim-sulfamethoxazole. Thus, infection with this bacterium should be considered in cases of antibiotic-insensitive fever with a rash, especially in young, old, and immunosuppressed persons. The organism is sensitive to doxycyclin, rifampicin, thiamphenicol, and fluoroquinolones (Clinicians encountering patients with fever or rash (or both) and a history of cat contact or flea bites should consider a diagnosis of


The fundamental need for environmental preservation has been calling, around the world, for the development and deployment of friendly industrial processes. Moreover, presently, development has to take into account sustainability. Within this context, enzyme technology is of outmost importance, having a major role for the achievement of the goals of sustainable development. This technology is of particular significance to Brazil due to the need to preserve Brazil's singular ecosystems, biodiversity, and quality and availability of the country's water resources. Furthermore, Brazil has an unrivalled availability of natural wealth, in both diversity and quantity, to be processed, via biocatalysis, into a wide range of diverse and innovative products as well as into biofules. Enzymes are the logical tool to process renewable resources as both of them have naturally evolved to match each other. It is now, therefore, time to develop dexterity and expand the use of enzymes in a fully efficient manner. As biochemical processes are cleaner than its chemical counterparts and generate higher-quality products, this shift would benefit the country's socioeconomic development. However, going from traditional chemistry into enzymatic processes is more than a technical and economical challenge, it is a change in the way we think technology—scientific and technical knowledge— and thoughtfulness are much needed. vis-à-vis the necessary move toward biocatalysis. Eight papers, which were selected from a wealth of submitted manuscripts, were accepted for publication in this special issue of Enzyme Research. They relate to fundamental areas of enzyme research and technology such as microbial screening, enzymes production and characterization, fine chemistry, biorefinery, and biofuels. The Brazilian Seminar on Enzyme Technology (ENZITEC) has been taking place every two years since 2003. Its main objective has been to further the knowledge on the wide range of industrial, technical, and specialty enzymes alongside promoting its economic viability. The event encompasses the worldwide interest in industrial enzyme research and uses Denise M. G. FreireDenise M. G. FreireElba P. S. BonElba P. S. BonGustavo Viniegra-GonzalezGustavo Viniegra-GonzalezFrancisco GirioFrancisco GirioRobert F. H. DekkerRobert F. H. Dekker


Ixodes ricinus (Acari: Ixodidae) in Europe, as well as in changes in the distribution within its prior endemic zones. This paper builds on published literature and unpublished expert opinion from the VBORNET network with the aim of reviewing the evidence for these changes in Europe and discusses the many climatic, ecological, landscape and anthropogenic drivers. These can be divided into those directly related to climatic change, contributing to an expansion in the tick’s geographic range at extremes of altitude in central Europe, and at extremes of latitude in Scandinavia; those related to changes in the distribution of tick hosts, particularly roe deer and other cervids; other ecological changes such as habitat connectivity and changes in land management; and finally, anthropogenically induced changes. These factors are strongly interlinked and often not well quantified. Although a change in climate plays an important role in certain geographic regions, for much of Europe it is non-climatic factors that are becoming increasingly important. How we manage habitats on a landscape scale, and the changes in the distribution and abundance of tick hosts are important considerations during our assessment and management of the public health risks associated with ticks and tick-borne disease issues in 21st century Europe. Better understanding and mapping of the spread of I. ricinus (and changes in its abundance) is, however, essential to assess the risk of the spread of infections transmitted by this vector species. Enhanced tick surveillance with harmonized approaches for comparison of data enabling the follow-up of trends at EU level will improve the messages on risk related to tick-borne diseases to policy makers, other stake holders and to the general public.Many factors are involved in determining the latitudinal and altitudinal spread of the important tick vector  Ixodes ricinus (Acari: Ixodidae) is a small hard tick that transmits a large variety of pathogens of medical and veterinary importance. Those commonly reported include Borrelia burgdorferi s.l. causing Lyme borreliosis, Anaplasma phagocytophilum causing human granulocytic anaplasmosis, Francisella tularensis causing tularaemia, Rickettsia helvetica and Rickettsia monacensis causing spotted fever rickettsiosis, Babesia divergens and Babesia microti responsible for babesiosis, Neoehrlichia mikurensis, tick-borne encephalitis virus, Louping ill virus and Tribec virus.I. ricinus, and/or changed activity that bring people into contact with ticks.The most prevalent tick-borne infection of humans in the northern hemisphere is Lyme borreliosis, whose incidence has increased in at least nine European countries over the last decade-3. One pI. ricinus is known to be changing in Europe, both at extremes of altitude and latitude, as well as within its prior range. The reasons for these changes are manifold, and this paper aims to review the driving forces for changes in the geographical distribution of this tick species in Europe. Before we consider these changes, however, it is useful to summarise the biological and ecological aspects of the life history of I. ricinus that underpin the impact of so many different drivers on its survival, seasonal and diel activity and distribution.The distribution of Each active life stage attaches to a single host and feeds on blood for a period of days before detaching and then moulting (larvae and nymphs) or producing eggs . Larvae I. ricinus quest for hosts using an ‘ambush’ technique whereby they climb up vegetation and wait for a host to brush past. During questing, the tick loses moisture so has to descend the vegetation into the litter/mat layer to rehydrate. Moving back into the litter/mat layer reduces the probability of coming into contact with a host and uses up energy stores so is detrimental to tick survival, the risk factors can be divided generally into 1) those directly related to climatic change , 2) those related to changes in the distribution of tick hosts (which may be a direct or indirect effect of human intervention), and 3) other ecological changes . Drivers of change in distribution are discussed under the following headings: climatic effects at high altitude and latitudes (including impacts of tick habitats and tick hosts), habitat patchiness/connectivity and urban green corridors, expansion of tick hosts, and anthropogenic-specific factors, as well as evidence of expansion to new territories. The challenges in overcoming lack of historical data to assess changes are also discussed.Following expert opinion from tick researchers within VBORNET, a network of entomologist and public health experts supported by the European Centre for Disease Prevention and Control, and a comprehensive literature review [Searches were made of PubMed Google Scholar and Science Direct, with search terms including, I. ricinus with increasing altitude, as documented from vertical transects carried out between 620–1270 m.a.s.l (metres above sea level) during the period 2002–2006 in a mountainous region in Czech Republic[et al.[There is a clear biological basis for the reduced abundance of Republic,31. The c[et al. and therc[et al.. There wI. ricinus survival. Studies in Switzerland[I. ricinus is reported up to 1450 m.a.s.l in the study sites in Switzerland. However, the density of nymphs differs between south-facing and north-facing slopes. Density decreased with altitude on south-facing slopes[I. ricinus on south-facing slopes[I. ricinus were more common on east and west facing slopes[Additional effects of microclimate variables and the diversity of ecological factors add to the complexity of the impact that altitude might have on the vertical limit of tzerland-36 have g slopes, but incg slopes. This hag slopes, and thiI. ricinus is clearly dynamic, as altitude acts principally as a determinant of climate, and therefore thresholds vary with latitude across Europe. In Italy, the tick becomes far less abundant above 1300 m.a.s.l[I. ricinus ticks have been found below 600 m.a.s.l[I. ricinus[The altitudinal limit of  m.a.s.l. Further m.a.s.l, 1560 m. m.a.s.l, 1080–12 m.a.s.l. At the  m.a.s.l. Of the . ricinus. With reI. ricinus from studies in Bosnia & Herzegovina, the Czech Republic and Slovakia. Evidence from Omeragic[I. ricinus in Bosnia & Herzegovina in the 1950s was <800 m.a.s.l, and this increased to 900 m.a.s.l by the 1960s, and to 1190 m.a.s.l by 2010. Similar historical data also exists for the Czech Republic, from two separate mountain ranges. In Sumava, the highest limit of tick survival in studies in 1957 was 700 m.a.s.l. By the 1990s, ticks were being found up to and above 700 m.a.s.l in areas previously thought to be tick free, and by 2001, this had increased to 1100 m.a.s.l[I. ricinus.There is good field-based evidence of an expansion in the altitudinal range of Omeragic suggests m.a.s.l. Similar m.a.s.l, the alt m.a.s.l,45. In S m.a.s.l. AlthougI. ricinus to higher altitudes was due to increased annual and seasonal temperatures and rainfall, and consequently an extended period to allow for tick development. Danielová et al.[I. ricinus populations. They suggest that increased temperatures during January and February will also influence tick host survival, providing ample small mammal hosts for immature stages. In Krkonose, Danielová et al.[Changes in climate over recent years have been recorded throughout Europe, including at high altitude in the Czech Republic. Daniel et al. suggest vá et al.,48 reporvá et al. reportedoN so far examined – Sarah Randolph unpublished data); large inter-annual differences in the length of the seasons and the abundance of ticks will still be expected.Increasing temperatures will have a disproportionate effect on tick populations under the cooler and limiting conditions at higher altitudes because of the non-linear positive relationship between rates of development and temperature,50. At tI. ricinus, exemplified by studies in Greece that report a lower altitudinal limit of 600 m.a.s.l.[At high altitudes, snow cover during winter may enhance tick survival by minimizing repeated freezing and thawing in litter layers  to spread to and inhabit previously inhospitable areas of the Alps and Scandinavia. Such deer movements have allowed I. ricinus to be transported northwards on the Scandinavian Peninsula, resulting in a significantly increased tick range during the last 30 years[I. ricinus[Climate cannot act alone; furthermore, survival. Jaensonl[et al.,56 sugge30 years. Further. ricinus. The mov. ricinus.I. ricinus habitat at high altitude, particularly changes in forest and wildlife management[I. ricinus. With respect to animals, there has been a significant increase and re-population of the Alps by roe deer since the 1940s[I. ricinus feeding on mammal species that inhabit traditionally unfavourable conditions for ticks  may suggest the possibility for alternative tick-host interaction cycles .In Italy there have been similar reports of increased temperature at high altitude since the 1980s, but other factors evidently contribute to suitability for nagement. For exahe 1940s. The recI. ricinus (Maria Kazimirova pers. comm.), with ticks found in new regions of Slovakia where populations of I. ricinus were previously unrecorded[I. ricinus at these altitudes. Furthermore, at newly colonised sites for ticks, the local small mammal population would assist in sustaining immature I. ricinus stages.Similar habitat and tick-host changes have been reported in Slovakia and the Czech Republic. In Slovakia, there is evidence over the last 20 years that areas of formerly cultivated land at high altitude have been left uncultivated, possibly offering additional habitat both for wild rodents and recorded. In the recorded. This inI. ricinus occurs in northern parts of the UK[With regard to the northern extent of the range of this tick, f the UK and DenmI. ricinus in Sweden[I. ricinus to spread so rapidly and extensively to many previously tick-free localities of Northern Sweden[Various studies have monitored and explained the northward spread of n Sweden,56,63-66n Sweden. The shiI. ricinus was commonly encountered in areas where the VP exceeded 180 days, but rarely where it remained below 160 days. There was also a reduced duration of snow cover. With fewer than 125 days of snow cover, I. ricinus was consistently present, but above 175 days of snow cover it was consistently absent[A generally warmer climate can exert an effect in ways already outlined above with respect to warming at high altitudes. y absent. A proloy absent,65.I. ricinus are not as clearly defined and need further study[I. ricinus distribution in Fennoscandia will expand as winter seasons become shorter and milder, VP increases and deciduous woodland expands[I. ricinus activity is expected in southernmost regions and abundance is expected to increase in most of the regions by 2071–2100[Similar sorts of abiotic and biotic factors may also apply to Norway, although the driving forces behind an observed 400km northward expansion of er study. Various expands,64,68,69071–2100.I. ricinus range expansion in some parts of Europe, is supported by the results of a questionnaire survey in the UK[I. ricinus. The reasons for this are largely anecdotal, but are thought to be due to a combination of events: a behavioural adaptation in roe deer in relation to their tolerance of human disturbance, ongoing afforestation of the country by 10-25%, changes in agricultural practice (including the feeding of deer), and a decline of predators (due to scabies in foxes). Similarly, their expansion to all regions may be limited by the occurrence of sandy soils in some regions of the country[The above argument, that deer expansion appears to be a major driving force for n the UK and fieln the UK, and dee country.I. ricinus populations, other vertebrate species may play that role in some localities. In certain parts of Germany, where roe deer populations have reportedly remained stable, large increases in populations of wild boar have been linked with increases in I. ricinus abundance[I. ricinus. On some small isolated islands in the Baltic Sea, such as Gotska Sandön and other small islands along the Fennoscandian coastline, cervids are absent, but the I. ricinus populations can be maintained by mountain hares (Lepus timidus) that are the only host for all active stages of the tick populations[I. ricinus[Although deer are commonly the most important wild animal in sustaining bundance. There aulations. In othe. ricinus.I. ricinus can survive and spread to new regions. While an extended VP and expansion of deciduous woodland is important in northern latitudes, there are also many afforestation and habitat connectivity initiatives within other parts of Europe, well within the prior range of I. ricinus. In Spain, there is evidence that long-term and short-term changes in climate suitability are enabling some areas of the country to become more favourable and others less favourable  . Habitat configuration, that is the degree of connectivity between habitats, is important in determining tick presence/absence and abundance through its effect on tick hosts[I. ricinus associated with greater connectivity between patches, any reduction in the distance between patches (thus promoting de-fragmentation of habitats) may promote the probability of successful invasion and establishment of I. ricinus in new areas[Habitat structure clearly plays an important part in determining whether ck hosts,74. Withew areas.I. ricinus. There appears to have been an expansion in the range of I. ricinus in south-west England during the last decade[I. ricinus are exploiting ‘field margins’ (i.e. margins of arable land left for wildlife), although the occurrence of ticks is influenced by the neighbouring habitat (e.g. woodland versus arable), and the local activities/movements and daily resting behaviours of deer. Woodlands are also being managed as mosaic habitats, with active management of track-side grassland (known in the UK as a ride) within forests generating compartmentalised woodland, specifically for the benefit of butterflies. This provides sunny path-side vegetation for a range of animals and plants. Various factors, such as leaf litter, sward height and occurrence of bracken and bramble, along with a favourable aspect (i.e. orientation) appear to favour tick abundance[In the UK and in other European countries, agri-environment schemes encourage de-fragmentation of habitats by providing habitat corridors between ‘patches’ of extant habitat. This provides a corridor for animal dispersal, refuges for their browsing and laying up (e.g. by deer) and consequently will affect the dispersal of t decade. This mabundance,76. In tbundance. Tick pobundance. NeverthI. ricinus is whether the species is a generalist, adapted to a very wide range of climate conditions, or exists as a series of dynamic populations in different regions within Europe, adapted to local prevailing conditions. The only phenotypic analysis on several populations of the species involved the use of cuticular hydrocarbon composition to characterize the chemical profile of different specimens[A further complication to the argument of dispersal and local establishment of pecimens,78. SuchI. ricinus in Europe lead to contradictory opinions; from the lack of genetic structure of the species in the target area[I. ricinus is linked to the type of host used, which in turn is an indicator of the type of habitat used by the species[Studies of allozymic data or DNA sequences of get area,80 to thget area, to a laget area. Some st species. In any I. ricinus is present throughout the country except at mid- and high altitude. There have been no specific studies tracking its spread, in common with almost all countries apart from those described above. It is, however, generally acknowledged that there has been an increase in abundance of I. ricinus in France . Natural, social and anthropogenic factors are generally accepted by French tick experts as contributing to a de-stabilisation of the previous status quo that may have led to an increase in tick abundance[I. ricinus in France are too fragmentary to make any firm assessments of species expansion. Similarly, many unsolicited casual reports from members of the general public, comparing the lack of a tick problem when they were young with the need to protect the current generation of children from exposure to ticks during recreation, all point to a more widespread distribution of ticks in the UK, especially in peri-domestic habitats . This is presumably due to a range of causes, some of which may have an anthropogenic originIn France, bundance. Much ofI. ricinus, but this does not allow a quantification of tick expansion, or indeed constitute evidence of species expansion. There are anthropogenic drivers in Portugal, such as changes in land management, changes in pest control strategies, extensive destruction of habitats by fire and an increase in hunting, all of which could contribute to any real change (both increase and decrease) in the abundance and distribution of I. ricinus[I. ricinus to central and northern parts of the country.In Portugal, the recent increase in studies on the epidemiology of tick-borne disease has led to a greater awareness of the occurrence and spatial distribution of . ricinus. Coupled. ricinus. The latIn central and eastern European countries, the reform of agricultural practices after the fall of Soviet rule has led to significant changes in land cover and land use, most of which act synergistically to improve the habitat for ticks and increase human contact with ticks. National herds of cattle and sheep have declined , and the area under cultivation of field crops has diminished, both allowing natural regeneration of herbaceous and woody vegetation and re-iI. ricinus, and in some cases we have field-based evidence for such effects. Often our opinion on whether ticks are increasing in their range is based largely on anecdotal evidence, which is often hard to quantify. Recently published maps from the UK[st century sources including: incidence data on Lyme borreliosis and bovine babesiosis, and observational data of tick occurrence from veterinary surgeons, from the public via media sources and from cervid hunters[I. ricinus is known to be patchy, although a Portuguese surveillance programme for ticks and tick-borne diseases of public health concern (REVIVE) was implemented from 2011[So far we have presented several theories on the driving forces for change in distribution of m the UK have comm the UK. A multi hunters. In Portrom 2011. It is eI. ricinus[Reliable, historical tick distribution data are valuable for assessing changes in the populations of . ricinus. Unfortu. ricinus. Possibl. ricinus and in m. ricinus.I. ricinus within countries, there is recent evidence of expansion of I. ricinus into new off-shore territories. The first record of I. ricinus in the Faroe Islands came from a dog in 1999. This was followed by records of a nymph on a wheatear in 2000, an engorged female on a cat in 2004, an engorged larva on a chiffchaff in 2004 and a nymph on a human in 2005 . Ticks I. ricinus distribution. In Estonia for example, there have been extensive collections of I. ricinus for pathogen detection since the 1980s. Although fluctuations in tick abundance were noted, no specific studies on changing abundance or distribution have been conducted . In Latvia, I. ricinus is considered largely absent from eastern parts of the country (where I. persulcatus is more common) and there is no evidence of recent spread . However the above-mentioned landscape changes (both newly re-generated woodlands and within existing forests) in Latvia could continue to affect tick abundance and distribution in the future.For some countries there is no reported evidence of an expansion in I. ricinus based on the publication records of the last 10 years and will update these maps based on a compilation of existing data from various sources provided and shared by the members of the network. This unique map resource will form the basis for the better documenting the range expansion of I. ricinus and other tick species relevant to public health.At the European level, VBORNET has started to map the distribution of I. ricinus as well as in changes in the distribution within its prior endemic zones. The drivers can be divided into those directly related to climatic change, those related to changes in the distribution of tick hosts particularly roe deer and other cervids, or other ecological changes and anthropogenically induced changes (TableI. ricinus (and changes in its abundance) are, however, essential to assess the risk of the spread of infections transmitted by this vector species. Enhanced tick surveillance with harmonised approaches will enable better comparisons to be made of change in trends in distribution at an EU level. This will help improve the messages on the public health risk of tick-borne diseases to policy makers, other stakeholders and to the general public.Many factors are involved in the latitudinal and altitudinal spread of es Table. These fThe author(s) declare that they have no competing interests.JMM and KMH conducted the literature review and were responsible with WVB for collating and writing the paper. All other authors provided expert information and comment relevant to their area of expertise and geographic region to ensure that the paper was not geographically biased. The VBORNET project is led by GH, in coordination with WVB and HZ. The work-package responsible for this review is coordinated by JMM. All authors read and approved the final version of the manuscript.


Yersinia. In this issue, the focus was on yersiniosis-related foodborne illnesses, behavior of Yersinia in foods, incidence, persistence, survival, or growth, outbreaks and surveillance, zoonosis virulence and pathogenesis, detection/identification, mechanisms to grow in foods, and public health. Yersinia belongs to the Enterobacteriaceae family and is often isolated from clinical specimens. Three Yersinia strains, namely, Y. enterocolitica, Y. pseudotuberculosis, and Y. pestis, are pathogenic to humans and are widespread among various animal species and in the environment. They are transmitted to humans by the oral route and cause intestinal symptoms such as abdominal pain, diarrhea, and fever. These species are found all over the world, with a higher incidence in temperate and cold environments. This special issue of Journal of Pathogens was designed to share some of the interested scientific studies published on yersiniosis, a foodborne outbreaks associated with consumption of food contaminated with Yersinia enterocolitica in foods, their incidences, possible route of contamination, persistence, factors that influence the survival, or growth in food, soil, and water are reviewed by Bari et al. In this special issue, behavior of Yersinia spp. and their current status in different foods and environments are discussed by A. Rahman et al. The epidemiology, outbreaks and surveillance, and zoonosis of Yersinia enterocolitica, mode of transmission of virulence, and their factors are covered by Y. Sabina et al. The molecular insight of virulence of Yersinia enterocolitica and Y. pseudotuberculosis in human yersiniosis, their genomics, mechanisms of infection, and host responses including the current state of surveillance, detection, and prevention of yersiniosis, are presented by C. L. Galindo et al. The pathogenesis of Yersinia species and procedure to monitor the presence of virulence plasmid in Y. Pestis during storage and a convenient culture method for monitoring the presence of virulent plasmid in food are discussed by S. Bhaduri and J. L. Smith. The virulence plasmid (pYV) associated with the expression of phenotypic virulent in pathogenic Y. enterocolitica from pig tonsils was discussed by M. Denis et al. A highly sensitive, specific, and accurate selective chromogenic culture plate method that has been developed for detecting pathogenic Y. enterocolitica and Y. pseudotuberculosis in foods. H. Fukushima et al. reviewed and discussed the commercially available conventional and PCR-based procedures for specific detection of pathogenic Yersinia in foods. J. Gui and I. R. Patel reviewed and discussed the recent advances in molecular technologies and their application in detecting pathogenic Yersinia frederiksenii and their genomic analysis, and they found that enzyme might be responsible for the transposition of this novel retron element. R. Das et al. reported in their research article the presence of a novel single-stranded DNA in Yersinia enterocolitica infections in foods.In the last reviewed article, S. N. Aziz and K. M. S. Aziz discussed the theoretical modeling to avoid exposure of Latiful BariLatiful BariDike O. UkukuDike O. UkukuKenji IsshikiKenji IsshikiRamesh C. RayRamesh C. RayDidier MontetDidier Montet


Gender-specific smoking cessation strategies have rarely been developed. Evidence of effectiveness of physical activity (PA) promotion and intervention in adjunct to smoking cessation programs is not strong. SPRINT study is a randomized controlled trial (RCT) designed to evaluate a counselling intervention on smoking cessation and PA delivered to women attending the Italian National Health System Cervical Cancer Screening Program. This paper presents study design and baseline characteristics of the study population.Among women undergoing the Pap examination in three study centres , participants were randomized to the smoking cessation counselling [S], the smoking cessation + PA counselling [S + PA], or the control [C] groups. The program under evaluation is a standard brief counselling on smoking cessation combined with a brief counselling on increasing PA, and was delivered in 2010. A questionnaire, administered before, after 6 months and 1 year from the intervention, was used to track behavioural changes in tobacco use and PA, and to record cessation rates in participants.p < 0.001), smoked more cigarettes per day , and were more likely to have already done 1 or more quit attempts . The approach of SPRINT study appeared suitable to enrol less educated women who usually smoke more and have more difficulties to quit.Out of the 5,657 women undergoing the Pap examination, 1,100 participants (55% of smokers) were randomized in 1 of the 3 study groups . The three arms did not differ on any demographic, PA, or tobacco-use characteristics. Recruited smokers were older, less educated than non-participant women, more motivated to quit (33% vs.9% in the Preparation stage, ISRCTN52660565ISRCTN:  Smoking is the leading cause of death and of many diseases for both men and women . Since 1The National Health System Cervical Cancer Screening Program (NHS-CCSP) in Italy is a beneficial setting to deliver smoking cessation counselling to women aged 25-64 years attending the Pap test examination. Smoking is an important co-factor for the development of cervical cancer, being human papilloma-virus (HPV) the principal causal factor . HPV-posThe linkage between smoking cessation and the fear to gain weight is well documented. Women's concern about their weight has been hypothesized to be a factor in preventing smoking cessation efforts, increasing smoking relapse rates among quitters, and encouraging smoking initiation. Women are more likely than men to believe that smoking helps control body weight and are more likely to report actually using cigarettes as a means to control their weight . BecauseThe SPRINT Study was designed in order to verify the effectiveness of a standard counselling intervention on smoking cessation delivered by trained midwives in a gender-specific setting , and whether the adjunct of PA counselling to the standard smoking cessation counselling might increase quit rates among women undergoing the NHS-CCSP. This paper describes the characteristics of the female smokers enrolled in the study in comparison to those registered in non-participant female smokers.The Sprint intervention is a standard brief counselling on smoking cessation combined with a brief counselling on increasing PA. The smoking cessation counselling corresponds to the first two phases of the brief intervention for smoking cessation ("Ask" and "Advice") . It is tIT is a three-arm randomized controlled trial. Smokers attending the NHS-CCSP consulting rooms who decided to participate in the study, were the units of analysis and were randomly assigned to one of the two intervention arms  or the control [C] arm Figure .The study population, consisting of women 25-64 years of age attending some NHS-CCSP consulting rooms, was expected to be enrolled in three Italian areas . Women waiting to perform the test were asked to fill in the study questionnaire and, if participating in the study, the informed consent.Assuming a six-month cessation rate in the S + PA arm of about 15%, and a cessation rate in the C group of about 8% (relative risk of 1.8), with a significant level α = 0.05, and a power of 0.80, a sample size of about 430 women per arm, and about 1,300 women overall, were estimated to be necessary to conduct the study. Cessation rates used in the power calculation derived from a pilot study previously conducted in one NHS-CCSP consulting room in Florence . KnowingAll smoking women, aged 25-64 years and attending the NHS-CCSP consulting rooms in the study areas, irrespective of their health status, were eligible if they consented to participate in the study. Participating women were randomly assigned to one of the 3 arms of the study by the midwives, using a predefined list of casual numbers.Participants had to fill in a questionnaire before to perform the test. Questions covered demographic characteristics, lifetime and current use of cigarettes, frequency of previous quit attempts, the Fagerstrom Tolerance Questionnaire (FTQ) , and intA telephone follow-up to all participants was scheduled after six months and one year from the intervention, in order to know how many women successfully quitted after the intervention. The same questionnaire for tobacco use and PA levels was used in the follow-up interviews, in order to assess the Di Clemente-Prochaska stages of change for smoking cessation and increasing PA after the intervention. The analysis of these data are ongoing.The SPRINT Study was submitted and approved by the Ethics Committee of the Local Health Authority of Florence, Italy.U test was used to test for difference in continuous variables, since data showed a non-normal distribution.We conducted a descriptive analysis of the baseline characteristics of non-participant female smokers vs. recruited female smokers and across the study arms and centres. Differences in proportions were analyzed using the Chi-squared test. The non-parametric Mann-Whitney A total of 5,647 women who underwent the Pap smear test filled in the study questionnaire; 2,002 of them (35.4%) were current smokers, and 1,100 of them (54.9% of smokers) decided to participate to the study. Then, 363, 366, and 371 participants were randomly assigned to the S, S + PA, and C arms, respectively , and were more likely to have already done ≥ 1 quit attempts than non-participants . Finally, looking at the Di Clemente-Prochaska stages of change for smoking cessation, recruited women were more likely to be in the preparation and contemplation stages, whereas non-participating women were more likely to be in the precontemplation stage ,  and 19.6The three study centres  recruited the expected study samples, and recorded a participation rate from 50% in Turin to 65% in Mantua, higher than the participation rate of 31.5% recorded in the pilot study previously conducted . This hiOther strengths of the study are the large sample size and the measurement of variations from baseline stage of self-reported behavioural change for smoking cessation and increasing PA after the intervention.Recruited women recorded peculiar tobacco-use characteristics in comparison to non-participant women: they were more motivated to quit (about one third was in the preparation stage), smoked more cigarettes per day, and had already attempted to quit. Eighty-one percent of smokers in the preparation stage for smoking cessation, 64% in the contemplation stage, and only 38% in the pre-contemplation stage participated voluntarily to the study. Thus, this distribution by motivation level among participants could determine an unexpectedly high cessation rate in the control group. Anyhow, this bias is unavoidable in intervention studies where the intervention cannot be imposed and the subjects under study must be necessarily volunteers.Another limit of the study is that all the outcome measures on smoking cessation and PA are self-reported, even though self-reports are considered a low-cost approach to obtaining sufficiently accurate information on tobacco use and PA ,28.Regarding the motivational phase, it must be outlined that for PA the percentage of sedentary women in pre-contemplation (those who does not want even to think to change their habit) is similar in recruited and in non participant women ; on the contrary pre-contemplating smokers among non participants are twice the participants (61.6% vs 30.7%). These data reflect the high attention of women on their body weight and at the same time the resistance to quit. Smoking appeared to attenuate weight gain over time and this is also a reason of female smokers' resistance to quit smoking, even if the average weight of women who are current smokers is modestly lower than that of women never smoked or who are former smokers -29.In conclusion, smoking is actually the predominant health problem in developed countries, accounting for about 19,000 attributable deaths in Italian women in 2010 . PromotiThe authors declare that they have no competing interests.EC conceived of the study, participated in the design of the study, coordinated the multicentric study group and draft the manuscript. GG participated in the design and analysis of the study, and contributed to the manuscript draft. GC analysed the data and contributed to the manuscript draft. LG participated in the design of the study, was responsible of the study in Piedmont, and contributed to the manuscript draft. EA participated in the design of the study, was responsible of the study in Lombardy, and contributed to the manuscript draft. AI participated in the design of the study, was responsible of the study in Tuscany, and contributed to the manuscript draft. CB participated in the design of the study and trained the personnel involved in recruitment. EG participated in the design of the study, trained the personnel involved in recruitment, and revised the manuscript. AC participated in the design of the study, carried out the study in Piedmont and revised the manuscript. FT participated in the design of the study, carried out the study in Lombardy and revised the manuscript. MRG participated in the design of the study, carried out the study in Tuscany and revised the manuscript. All authors read and approved the final manuscript.The SPRINT Working Group includes the previous mentioned authors and all the midwives who enrolled the population under study: E. Amadori ; E. Anghinoni ; A.M. Badiali ; B. Baldini ; B. Baluga ; A.M. Barbi ; M.C. Barbieri ; C. Bellati ; B. Benatti ; E. Berini ; L. Boldrini ; P. Breviglieri ; C. Bronchi ; V. Cacciarini ; L. Calabrese ; L. Campitelli ; L. Caraffa ; N. Casoni ; G. Catelani ; D. Casi ; P. Cavini ; E. Chellini ; S. Clara ; A. Cerchi ; N. Colledan ; A. Coppo ; L. Cozzi ; C. Danielis ; C. Di Pierro ; F. Di Stefano ; C. Ferrari ; M. Ferri ; T. Ferri ; L. Fiaccadori ; F. Florio ; C. Forlucci ; E. Galanti ; M. Gialdini ; L. Giordano ; M.R. Giovacchini ; G. Gorini ; E. Grechi ; L. Grossi ; S. Guzzo ; G. Innocenti ; A. Iossa ; S. Labardi ; C. Lepri ; D. Montovanelli ; G. Mantovani ; A. Marchi ; S. Mazzoni ; M. Molinari ; M. Morandini ; R. Nidiaci ; P. Noli ; A.M. Notarangelo ; S. Oliveri Del Castillo ; R. Pasini ; F. Perfetti ; C. Prati ; G. Pria ; S. Raffanini ; N. Rigoni ; E. Russo ; R. Sgarbi ; M.E. Siliprandi ; D. Simoncelli ; F. Talassi ; C. Ticci ; S. Toffalini ; C. Tovagliari ; M. Vaccari ; C. Vair , M. Zambello .The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/11/906/prepub


Thermomonospora curvata Henssen 1957 is the type species of the genus Thermomonospora. This genus is of interest because members of this clade are sources of new antibiotics, enzymes, and products with pharmacological activity. In addition, members of this genus participate in the active degradation of cellulose. This is the first complete genome sequence of a member of the family Thermomonosporaceae. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 5,639,016 bp long genome with its 4,985 protein-coding and 76 RNA genes is a part of the  Genomic  Encyclopedia of  Bacteria and  Archaea  project. T (= DSM 43183 = ATCC 19995 = JCM 3096) is the type strain of Thermomonospora curvata, which in turn is the type species of the genus Thermomonospora . Thrmed 4B2 . T. curv type IV ,45 are,  Genomic  Encyclopedia of  Bacteria and  Archaea  project [This organism was selected for sequencing on the basis of its phylogenetic position , and is  project . The gen project  and the T. curvata B9T, DSM 43183, was grown in DSMZ medium 550  [et al. [l, 1973)  at 45°C. [et al. . DNA is  [et al. ,50.The genome of was sequenced using a combination of Sanger and 454 sequencing platforms. All general aspects of library construction and sequencing can be found at the JGI website . PyroseqGenes were identified using Prodigal  as part The genome consists of a 5,639,016 bp long chromosome with a 71.6% GC content . Of the