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Data collection and processing in the Azores was funded by Fundação para a Ciência e a Tecnologia (FCT) and Fundo Regional da Ciência e Tecnologia (FRCT), through research projects TRACE-PTDC/MAR/74071/2006 and MAPCET-M2.1.2/F/012/2011 (FEDER, the Competitiveness Factors Operational (COMPETE), QREN European Social Fund, and Pro convergencia Açores/EU Program). We also thank FCT for supporting MARE (UID/MAR/04292/2019) and OKEANOS (UIB/05634/2020), as well as for the research grants awarded to PR (SFRH/BPD/108007/2015) and CI (Project Awareness - PTDC/BIA-BMA/30514/2017). SMA is supported through project SUMMER (H2020-EU.3.2.3.1, GA 817806). Data collection by SECAC was funded by the EU LIFE programme—project LIFE INDEMARES (LIFE 07/NAT/E/000732)— and the Fundación Biodiversidad under the Spanish Ministry of Environment, Rural and Marine Affairs (project ZEC-TURSIOPS). EP was supported by a LLP/Erasmus grant 2010–2011 for collecting data in the Canary Islands. Background: Prioritizing groupings of organisms or ‘units’ below the species level is a critical issue for conservation purposes. Several techniques encompassing different time-frames, from genetics to ecological markers, have been considered to evaluate existing biological diversity at a sufficient temporal resolution to define conservation units. Given that acoustic signals are expressions of phenotypic diversity, their analysis may provide crucial information on current differentiation patterns within species. Here, we tested whether differences previously delineated within dolphin species based on i) geographic isolation, ii) genetics regardless isolation, and iii) habitat, regardless isolation and genetics, can be detected through acoustic monitoring. Recordings collected from 104 acoustic encounters of Stenella coeruleoalba, Delphinus delphis and Tursiops truncatus in the Azores, Canary Islands, the Alboran Sea and the Western Mediterranean basin between 1996 and 2012 were analyzed. The acoustic structure of communication signals was evaluated by analyzing parameters of whistles in relation to the known genetic and habitat-driven population structure. Results: Recordings from the Atlantic and Mediterranean were accurately assigned to their respective basins of origin through Discriminant Function Analysis, with a minimum 83.8% and a maximum 93.8% classification rate. A parallel pattern between divergence in acoustic features and in the genetic and ecological traits within the basins was highlighted through Random Forest analysis. Although it is not yet possible to establish a causal link between each driver and acoustic differences between basins, we showed that signal variation reflects fine-scale diversity and may be used as a proxy for recognizing discrete units. Conclusion: We recommend that acoustic analysis be included in assessments of delphinid population structure, together with genetics and ecological tracer analysis. This cost-efficient non-invasive method can be applied to uncover distinctiveness and local adaptation in other wide-ranging marine species. Publisher PDF

Data on Risso’s dolphin are very scarce, possibly due to its wide-ranging movements in the Mediterranean basin. Available data suggest the presence of this species in deep waters, shelf-break areas where the slope is steepest, around offshore islands and archipelagos. A total of eight sightings were recorded during vessel-based surveys carried out along opportunistic line transects from March 2017 to October 2019. The results of the monitoring activity showed that the occurrence of this species in the water surrounding Samos Island is unpredictable and that the characteristics of the environment of each sighting are various, as already shown by previous Mediterranean studies. Nonetheless, the results of this study are promising concerning Risso’s dolphin presence and conservation in the waters surrounding Samos Island.

The Marine Strategy Framework Directive (MSFD) aims to protect the marine environment and marine biodiversity across Europe more effectively. In the present study, the predicted distribution of striped dolphins of the Gulf of Corinth (Greece) is investigated for the first two periods of the MSFD (I: 2007-2012; II: 2013-2018). For the first time, anthropogenic variables are included in the striped dolphin distribution analysis, resulting from accounting for most of the variability. The results show that the predicted distribution is similar for both investigated periods. However, a small reduction of suitable areas in the Northern portion of the Gulf is observable for the second one, probably due to the expansion of fish farm activity in this zone. The MSFD and its consequent management actions didn’t appear to improve the protection of striped dolphins in the investigated area. Their review will be essential to assess which further management actions will be necessary to program to protect and conserve striped dolphins in the Natura 2000 area “Gulf of Corinth”. The present study shows that human activities are important for explaining stripe dolphins distribution and that their further development needs to be carefully taken into account while managing spatial planning.

Loggerhead turtles (Caretta caretta) spend most of their life in large marine areas occupying a variety of habitats where they are exposed to different types of threats. Among these, marine litter is known to pose a risk of entanglement or ingestion. Areas of risk exposure can be identified where the species overlap with litter accumulations, but gathering data on this highly mobile species and marine litter, especially in high sea areas, is challenging. Here we analysed five years of sea turtle and marine litter data collected by a network of research bodies along fixed trans-border transects in the Mediterranean Sea. Ferries were used as observation platforms to gather systematic data on a seasonal basis using standard protocols. The Sightings Per Unit of Effort was used as an index to compare loggerhead turtle sightings over time and space, and risk exposure areas were assessed based on seasonal overlap of species hot spots and high-density plastic areas revealed by Kernel analysis. In almost 180,000 km surveyed, 1258 sea turtles were recorded, concentrated mostly during all seasons in the central Adriatic Sea and Sardinia-Sicilian channels, and during spring in central Tyrrhenian. Plastic was the highest fraction of litter items detected. Several areas of higher risk exposure, both permanent and seasonal, were identified, mainly in the Adriatic Sea and during the spring-summer seasons. Records of both species and floating litter were highly variable, underlying the need for continuous long-term monitoring to develop sound conservation and management measures, especially in the identified areas of risk exposure.

Background The Mediterranean subpopulation of fin whale Balaenoptera physalus (Linnaeus, 1758) has recently been listed as Vulnerable by the IUCN Red List of threatened species. The species is also listed as species in need of strict protection under the Habitat Directive and is one of the indicators for the assessment of Good Environmental Status under the MSFD. Reference values on population abundance and trends are needed in order to set the threshold values and to assess the conservation status of the population. Methods Yearly summer monitoring using ferries as platform of opportunity was performed since 2008 within the framework of the FLT Med Network. Data were collected along several fixed transects crossing the Western Mediterranean basin and the Adriatic and Ionian region. Species presence, expressed by density recorded along the sampled transects, was inspected for assessing interannual variability together with group size. Generalized Additive Models were used to describe density trends over a 11 years’ period (2008–2018). A spatial multi-scale approach was used to highlight intra-basin differences in species presence and distribution during the years. Results Summer presence of fin whales in the western Mediterranean area showed a strong interannual variability, characterized by the alternance of rich and poor years. Small and large groups of fin whales were sighted only during rich years, confirming the favorable feeding condition influencing species presence. Trends highlighted by the GAM can be summarized as positive from 2008 to 2013, and slightly negative from 2014 to 2018. The sub-areas analysis showed a similar pattern, but with a more stable trend during the second period in the Pelagos Sanctuary sub-area, and a negative one in the other two sub-areas. Our findings further confirm the need for an integrated approach foreseeing both, large scale surveys and yearly monitoring at different spatial scales to correct and interpret the basin wide abundance estimates, and to correlate spatial and temporal trends with the ecological and anthropogenic drivers.

The Mediterranean endemic fan mussel Pinna nobilis is suffering an ongoing basin-scale mass mortality event (MME) since 2016. As most Mediterranean populations have collapsed, the species has been declared as Critically Endangered in the IUCN Red List of threatened species. In an effort to track the progress of the MME and provide updated information on the status of the species in the Greek seas, we compiled data collected through dedicated surveys and opportunistic assessments during 2019 and 2020. A total of 14589 fan mussel individuals, of which 81.1% dead, were recorded in 258 site surveys. Of the remaining 2762 live individuals, 256 were juveniles. Two marine areas which still sustain living populations were identified, namely Kalloni Gulf (Lesvos Island), and Laganas Bay (Zakynthos Island). The inner part of Kalloni Gulf appears to maintain the largest surviving population of the species in the eastern Mediterranean, with an abundance estimate of 684000 individuals (95% confidence interval: 322000-1453000). Solitary, potentially resistant, scattered individuals were recorded in several sites. Other previously abundant populations that had been assessed in the past, specifically those of Lake Vouliagmeni (Korinthiakos Gulf), Souda Bay (Crete) and Gera Gulf (Lesvos Island), and which collectively summed up to ~350,000 individuals, have now been wiped out. Our results document the collapse of most P. nobilis populations throughout the Greek seas. The MME has substantially progressed between early 2019 and mid-2020, as indicated by the increase of mortality at sites consecutively monitored multiple times. This work highlights the urgent need for continuous monitoring of surviving populations and calls for the immediate implementation of an effective protection and management strategy that will ensure the persistence of surviving individuals and the production of resistant offspring.

We are grateful to the ACCOBAMS Secretariat for their support in this work, including a small grant for the analysis. The Mediterranean population of Cuvieŕs beaked whale (Ziphius cavirostris), a deep-diving cetacean, is genetically distinct from the Atlantic, and subject to a number of conservation threats, in particular underwater noise. It is also cryptic at the surface and relatively rare, so obtain robust knowledge on distribution and abundance presents unique challenges. Here we use multiplatform and multiyear survey data to analyse the distribution and abundance of this species across the Mediterranean Sea. We use a novel approach combining heterogeneous data gathered with different methods to obtain a single density index for the region. A total of 594,996 km of survey effort and 507 sightings of Cuvier’s beaked whales, from 1990 to 2016, were pooled together from 24 different sources. Data were divided into twelve major groups according to platform height, speed and sea state. Both availability bias and effective strip width were calculated from the sightings with available perpendicular distance data. This was extrapolated to the rest of the sightings for each of the twelve groups. Habitat preference models were fitted into a GAM framework using counts of groups as a response variable with the effective searched area as an offset. Depth, coefficient of variation of depth, longitude and marine regions (as defined by the International Hydrographic Organization) were identified as important predictors. Predicted abundance of groups per grid cell were multiplied by mean group size to obtain a prediction of the abundance of animals. A total abundance of 5799 (CV = 24.0%) animals was estimated for the whole Mediterranean basin. The Alborán Sea, Ligurian Sea, Hellenic Trench, southern Adriatic Sea and eastern Ionian Sea were identified as being the main hot spots in the region. It is important to urge that the relevant stakeholders incorporate this information in the planning and execution of high risk activities in these high-risk areas. Postprint

To date, the principal assessment for the Loggerhead sea turtle (Caretta caretta) distribution in the Mediterranean Basin is given by beaches nests along the coasts. The Eastern Mediterranean Sea has been revealed an important foraging area for this protected species but temporal trends of its spatial distribution and anthropogenic threats are lacking. This study shows that the employment of ferry as platform of observation would represent a valid tool for monitoring Loggerhead sea turtle in remote marine area, and to pinpoint important areas for the conservation and assessment of the species. The results of the present study confirm the previous hypothesis of the areas selected by the species, which are neritic areas in the northern/middle Adriatic Sea and oceanic areas in the Ionian Sea.

The striped dolphin (Stenella coeruleoalba) is a cosmopolitan species common worldwide in tropical and temperate pelagic waters. The conservation status of the Mediterranean striped dolphin is defined as "vulnerable" [1]. Despite its large diffusion, the behaviour of striped dolphin is still not sufficiently known. The investigation of striped dolphin behaviour and its interaction with human activity in a protected area, as the Gulf of Corinth (Greece), would facilitate implementing measure for its conservation. The application of traditional behavioural sampling methods, the development of a dedicated ethogram and the use and customization of the software BORIS for sampling striped dolphin surface behaviour, from land and at sea, is discussed in the present manuscript. The description of behaviour observed for the first time and the difference among behaviours observed from land and at sea and in presence/absence of boat traffic is also presented. The results of this study show that measured behaviours can be considered sensors to monitor the effect of human activity, as boat traffic, on a cetacean’s population.

The relationship between a species and its habitat is complex but it is essential to study the distribution and habitat use of a protected species in order to identify regions where human activities and disturbances are likely to impact its population. The Marine Strategy Framework Directive informs that studies focusing on spatial and temporal species distribution and ecosystem variations (including links between habitats and species of marine mammals) are recommended to achieve Good Environmental Status (GES) of the EU’s marine water by 2020. However, the current level of information on the spatial-temporal distribution of marine mammals is often lacking. The striped dolphin is the most abundant cetacean of the Mediterranean Sea; nevertheless its Mediterranean population is listed as "vulnerable" by IUCN experts. In order to investigate the spatial distribution of striped dolphin within the Adriatic-Ionian Region, a cooperative project among different research groups was set up and a multi-source data approach was employed. Presence data collected during monitoring activity carried out from 2008 to 2018 were analysed by means of Maxent. Main topographic variables as bathymetry, slope and distance from coastline were considered and five distribution models were performed to highlight suitable areas for striped dolphin within the entire study area. Models are presented and discussed. The results of this study show that to investigate the distribution of a species at a regional scale it is necessary to monitor area with different environmental characteristics. In this regards, cooperative regional projects represent an opportunity for pooling together data that taken singularly would not necessarily reflect the distribution of species at a larger scale, but that analysed together would allow to acquire information on suitable habitats at a regional level.

Marine litter is a major source of pollution in the Mediterranean basin, but despite legislative requirements, scant information is available for the ongoing assessment of this threat. Using higher size classes as proxy for litter distribution, this study gave a synoptic estimation of the amount, composition, and distribution of floating macro-litter in the Mediterranean. The average amount of macro-litter was in a range of 2-5 items/km2, with the highest in the Adriatic basin. Seasonal patterns were present in almost all study areas and were significant in the Ligurian Sea, Sardinian-Balearic basin, and Central Tyrrhenian Sea. Plastic accounted for > 80% of litter in all areas and seasons, with the highest proportion in the Adriatic Sea, Ligurian Sea, and Sicilian-Sardinian Channels; in the Bonifacio Strait, Tyrrhenian Sea, and Sardinian-Balearic basin, litter composition was instead more diverse. Spatial analysis suggested an almost homogeneous distribution of litter without evident regular aggregation zones.

Acoustic observation can complement visual observation to more effectively monitor occurrence and distribution of marine mammals. For effective acoustic censuses, calibration methods must be determined by joint visual and acoustic studies. Research is still needed in the field of acoustic species identification, particularly for smaller odontocetes. From 1994 to 2012, whistles of four odontocete species were recorded in different areas of the Mediterranean Sea to determine how reliably these vocalizations can be classified to species. Recordings were attributed to species by simultaneous visual observation. The results of this study highlight that the frequency parameters, which are linked to physical features of animals, show lower variability than modulation parameters, which are likely to be more dependent on complex eco-ethological contexts. For all the studied species, minimum and maximum frequencies were linearly correlated with body size. DFA and Classification Tree Analysis (CART) show that these parameters were the most important for classifying species; however, both statistical methods highlighted the need for combining them with the number of contour minima and contour maxima for correct classification. Generally, DFA and CART results reflected both phylogenetic distance (especially for common and striped dolphins) and the size of the species.

To improve our understanding of the complex genetic and ecological structure of bottlenose dolphin (Tursiops truncatus) populations, we examined the acoustic features of communication signals from two geographically contiguous areas: the Central–Eastern North Atlantic and the Mediterranean Sea. Variations in the whistles were evaluated for four locations. Ten signal parameters were measured and used to statistically differentiate between the areas. Over 79 % of sightings were correctly classified by discriminant function analysis, confirming an acoustic differentiation between the two basins. The results of cluster analysis using the mean values of the parameters for each sighting showed that the three easternmost sightings from the Mediterranean and one sighting from the Canary archipelago formed a separate cluster from the rest of the Atlantic. The two sightings from the Alboran Sea in the west Mediterranean were grouped with the Atlantic recordings. There was more variability in whistles from the Atlantic Ocean consistent with data from genetic and photo-identification studies that document resident and non-resident animals in the area. The results suggest that the Alboran area may be inhabited by animals differentiated from the rest of the Mediterranean basin as a result of habitat features.

Genetic studies have shown that there are small but significant differences between the short-beaked common dolphin populations in the Atlantic Ocean and those in the Mediterranean Sea. The short-beaked common dolphin is a highly vocal species with a wide sound production repertoire including whistles. Whistles are continuous, narrowband, frequency-modulated signals that can show geographic variation in dolphin species. This study tests whether the differences, highlighted by genetic studies, are recognisable in the acoustic features of short-beaked common dolphin’s whistles in the two adjacent areas of the Atlantic Ocean and the Mediterranean Sea. From a selected sample of good quality whistles (514 recorded in the Atlantic and 193 in the Mediterranean) 10 parameters of duration, frequency and frequency modulation were measured. Comparing data among basins, differences were found for duration and all frequency parameters except for minimum frequency. Modulation parameters showed the highest coefficient o...

The striped dolphin is a cosmopolitan species distributed worldwide. Morphological and genetic studies strongly suggest that the Mediterranean and eastern North Atlantic populations are isolated from each other. The Mediterranean population is considered a distinct conservation unit by International Union for the Conservation of Nature experts, classified as "vulnerable." This study describes the geographical variation of the striped dolphin whistles within the Mediterranean Sea. Recordings were collected from 1996 to 2003 throughout the basin, employing multiple platforms. Thirty-seven independent sightings with acoustic data collection were made, and 599 whistles were extracted and considered for statistical analysis. Whistle analysis enabled the identification of sub-populations of striped dolphins within the Mediterranean Sea. The acoustic diversity observed reflects the genetic differences recently found among striped dolphins inhabiting different Mediterranean regions. The results of this study support the hypothesis that gene flow reduction plays an important role in determining variation in whistle duration and frequency parameters, while ecological and social factors influence parameters of the modulation domains. The ability to acoustically identify distinct geographic sub-populations could provide a useful tool for the management of this protected species.

Geographic variation in the acoustic features of whistles emitted by the striped dolphin (Stenella coeruleoalba) from the Atlantic Ocean (Azores and Canary Islands) and the Mediterranean was investigated. Ten parameters (signal duration, beginning, end, minimum and maximum frequency, the number of inflection points, of steps, of minima and maxima in the contour and the frequency range) were extracted from each whistle. Discriminant function analysis correctly classified 73% of sounds between Atlantic Ocean and Mediterranean Sea. A cline in parameters was apparent from the Azores to the Mediterranean, with a major difference between the Canaries and the Mediterranean than between Azores and Canaries. Signal duration, maximum frequency, and frequency range measured in the Mediterranean sample were significantly lower compared to those measured in the Atlantic. Modulation parameters played a considerable role in area discrimination and were the only parameters contributing to highlight the differences within the Atlantic Ocean. Results suggest that the acoustic features constrained by structural phenotype, such as whistle's frequency parameters, have a major effect on the Atlantic and Mediterranean separation while behavioral context, social, and physical environment may be among the main factors contributing to local distinctiveness of Atlantic areas. These results have potential passive acoustic monitoring applications.

One goal of the LIFE project “Del.Ta.” (NAT/IT/000163) was the preparation of an Action Plan to protect the bottlenose dolphin community in the Pelagie Archipelago (Sicily, Italy). It stressed the importance of regular monitoring of the spatial and temporal distribution of dolphins in order to evaluate the impact of local activities. This study assesses whether land-based surveys could be an effective alternative to vessel-based surveys. During the summer of 2006, both surveys’ methodologies were used at Lampedusa, with 35 sightings recorded from land and 31 from a boat. Comparison was based on the assessment of the type of information they provided in relation to the presence of the animals and their behavior. Both methodologies were applicable, but there were differences in their requirements, potential information generated, costs, and sensitivity to weather conditions. Vessel-based surveys require well trained observers and enable photo-identification and observation of social interaction and morphology. Animal movements, interactions with anthropogenic elements and group dynamics are better collected from land but spatial data can be documented up to 1 nautical mile from the coast. Weather conditions have a significant platform specific effect on sighting frequencies. The high sighting frequency during land surveys provides support for the development of zero-impact land-based dolphins watching activity.

Acoustic signals are expressions of phenotypic diversity and their variation could provide important information on differentiation patterns within species. Due to a number of selective pressures acting on signals, the contribution of genetic drift is often complex to outline. This study aims at evaluating if an examination of the acoustic structure of communication signals can allow the identification of evolutionary units in species capable of vocal learning. We quantified and compared parameters of whistles emitted by three dolphin species (Stenella coeruleoalba, Delphinus delphis, and Tursiops truncatus) to examine the hypothesis that acoustic signals permit the recognition of differentiation between populations from the Atlantic Ocean and the Mediterranean Sea. In the three species, recordings were correctly assigned to their basin of origin with a percentage higher than 82% by DFA. Frequency parameters were the most stable within each species. Where gene flow has been shown, i.e., within Atlantic Oc...

Exhaled nitiric oxide (NO) is increased in exhaled breath of asthmatic patients. The aim of this study was to investigate the longitudinal changes of exhaled NO outside and during the pollen season in pollen-allergic asthmatic children. Twenty-one children (age 6 to 16 yr), with a seasonal allergic asthma sensitive to grass pollen, underwent measurements of exhaled NO and pulmonary function before (March), during (May), and after (November) the pollen season. Exhaled NO was measured by a tidal breathing method with a chemiluminescence analyzer and NO steady-state levels were recorded. The timing of the measurements during the pollen season was based on the atmospheric pollen count. Exhaled NO values of asthmatic children were compared with those of 21 sex- and age-matched healthy children. Pulmonary function and symptoms of asthma were also evaluated at each visit. The mean value of exhaled NO before the grass season was 12.7 ± 5.1 ppb (mean ± SD), significantly higher when compared with controls (7.8 ± 2...

Nitric oxide (NO) can be detected in human exhaled air, and its endogenous production is increased in patients with asthma. It may provide a noninvasive means for measuring airway inflammation. The aim of this study was to establish reference values for exhaled NO concentrations in a large number of healthy school-age children. We measured exhaled NO levels in 159 white healthy children (88 girls, 71 boys, age range 6-15 years) recruited from two public schools of Padua, Italy. Exhaled NO levels in exhaled gas were measured by a tidal breathing method with a chemiluminescence analyzer, and NO steady-state levels were recorded. Nasal NO levels were measured by direct sampling from the nose during mouth breathing. The mean concentration of endogenous NO in orally exhaled gas was 8.7 parts per billion (ppb) (95% confidence interval (C.I.), 8.1-9.2 ppb) and sampled data followed a log-normal distribution (Kolmogorov-Smirnov d = 0.77, P > 0.2). No difference was found between boys (mean value, 8.4 ppb; 95% C.I., 7.3-9.4 ppb) and girls (mean value, 8.9 ppb; 95% C.I., 7.9-9.9 ppb). No significant correlation was found between age, height, or spirometric data and exhaled NO levels (r < 0.2). The mean value of nasal NO concentrations was 216 ppb (95% C.I., 204-228 ppb). There was no correlation between exhaled and nasal NO values (r = 0.16, P = ns). In conclusion, this study establishes a reference range for exhaled NO values measured by a tidal breathing method in children between age 6-15 years. The observed levels are independent of age, gender, and lung function, and can be used to monitor airway inflammation in asthmatic children.

It has been hypothesized that concentrations of exhaled nitric oxide (NO) may be related to the extent of cytokine-mediated airway inflammation. Recent findings indicate the nasal airways as an important site of NO production. Our objective was to evaluate whether children with allergic rhinitis show different nasal NO levels when compared with normal healthy subjects and the effect of topical steroids and anti-histamine therapy. We have measured the concentration of NO drawn from the nose of 21 children (5-17 years old) affected by perennial allergic rhinitis (house dust mite) out of therapy for at least 3 weeks. Thirteen children were then treated with nasal beclomethasone dipropionate (BDP) (400 micrograms daily) and eight subjects with nasal anti-histamine levocabastine (200 micrograms daily). Measurements were performed before and after 10 days of treatment. As a control group we evaluated 21 healthy children aged 5-15 years. To measure NO we used a chemiluminescence analyser. Before treatment the whole group of children with allergic rhinitis showed a mean (+/- SEM) nasal NO concentration of 267 +/- 18 ppb, significantly higher (P < 0.01) than the control group (186 +/- 15 ppb). The group of children treated with BDP showed, after 10 days of therapy, a significant (P < 0.05) decrease of nasal NO concentration (271 +/- 21 ppb vs. 212 +/- 20 ppb). Indeed, in the group treated with levocabastine, nasal NO concentrations did not present a significant difference (P not significant) compared with baseline (261 +/- 33 ppb and 252 +/- 31 ppb, respectively). These data suggest that (1) children with allergic rhinitis have higher levels of nasal NO than non-atopic controls and (2) intranasal steroid therapy significantly reduces nasal NO production in children with allergic rhinitis. We speculate that the allergic inflammatory response may influence the nasal NO levels and that NO measurements may be a useful marker of nasal inflammation.

Recently, it has been demonstrated that paranasal sinuses are an important site of nitric oxide (NO) production in the upper airways. The aim of this study was to evaluate the NO nasal concentration in children with acute maxillary sinusitis before and after treatment with antibiotic therapy. We performed NO nasal measurements in 16 children 4 to 13 yr of age with acute maxillary sinusitis and compared values with 16 age- and sex-matched healthy control subjects. The diagnosis of acute sinusitis was done by clinical signs and symptoms in addition to radiographic examination. NO nasal concentrations were measured by a chemiluminescence analyzer. Nasal NO steady state during oral breathing was recorded. The mean +/- SEM NO nasal concentration in children with sinusitis was 70 +/- 8.7 parts per billion (ppb) and increased significantly to 220 +/- 15 ppb (p0.001) after antibiotic therapy (amoxicillin/clavulanate). NO values after recovery from sinusitis were similar to those of healthy control subjects (245 +/- 15 ppb, p = NS). NO nasal measurements were also performed before and after antibiotic treatment in nine children 4 to 12 yr of age with symptoms of upper respiratory tract infection but no symptoms of sinusitis. In these children NO nasal levels were 249 +/- 32 ppb and did not change (p = NS) after antibiotic therapy. We conclude that during acute maxillary sinusitis the concentration of nasal NO is largely decreased, probably because of an impaired flow of NO from the paranasal sinuses, and that NO returns to normal levels after antibiotic therapy.

Since 2003, as part of the LIFE project ‘Del.Ta.’ (NAT/IT/000163) a bottlenose dolphin community has been studied in order to assess the impact of increasing boat traffic in the Pelagie Archipelago (Sicily, Italy). During the summer of 2006 and 2008, land-based surveys were carried out at Lampedusa, one of the Archipelago's Islands, at set hours from 6 locations (7 in 2008) in standard weather conditions. Continuous horizon scanning and focal group sampling methods were used to study dolphin presence and behaviour. Dolphin–boat interactions were recorded for each sighting. A total of approximately 559 hours were spent monitoring and 83 sightings were recorded. With regard to the different types of vessels, interaction with sailing boats was always neutral, whereas fast boats caused the interruption of all activities and avoidance behaviour. Positive interaction occasionally occurred with bottom trawling fishing boats. Interactions recorded when fishing and powered engine boats approached within 200 m predominantly resulted in a negative reaction from the dolphins (approximately 70%) and dolphin behaviour was affected in terms of occurrence and patterns. Behavioural analysis showed that the animals reduced the frequency of behaviours related to feeding, social and relaxing activities. The dolphins also increased information-gathering behaviour such as spyhopping. The use of Markovian chain models revealed evidence of disruptions in sub-sequences made up of behaviours often associated with feeding. After boat departures, the animals rarely restarted their previous activity and persistence of this effect could have long-term consequences as a result of non-habituation. Sighting duration also decreased in the presence of vessels. The possible negative long-term effect of the presence of boats may be indicated by the lower mean sighting duration in 2008, when the highest presence of powered engine boats was recorded. In view of the scenario outlined above, local authorities need to enforce the requirement of maintaining a distance of over 200 m away from the animals for all types of boat and also promote regular land-based surveys of vessel impacts for tourist boats and dolphin watching cruises.

Exhaled nitric oxide (ENO) has been proposed as a marker of airway inflammation in asthma and could be useful to evaluate the response to anti-inflammatory treatment. We investigated the effect of budesonide and nedocromil sodium on ENO levels and lung function in asthmatic children. Twenty stable steroid-naı̈ve asthmatic children were randomized in a single blind, cross-over study to receive inhaled budesonide (group A) or nedocromil sodium (group B) for 6 weeks. ENO was measured with a chemiluminescence analyser at baseline and at the end of each treatment period. Repeated-measures ANOVA was carried out. In asthmatic baseline ENO levels [mean 32·5 ppb, 95% confidence interval (CI) 26·4 to 38·7] were significantly higher compared to reference values (8·7 ppb, 95% CI 8·1 to 9·2,P

Nitric oxide (NO) can be detected in human exhaled air, and its endogenous production is increased in patients with asthma. It may provide a noninvasive means for measuring airway inflammation. The aim of this study was to establish reference values for exhaled NO concentrations in a large number of healthy school-age children. We measured exhaled NO levels in 159 white healthy children (88 girls, 71 boys, age range 6-15 years) recruited from two public schools of Padua, Italy. Exhaled NO levels in exhaled gas were measured by a tidal breathing method with a chemiluminescence analyzer, and NO steady-state levels were recorded. Nasal NO levels were measured by direct sampling from the nose during mouth breathing. The mean concentration of endogenous NO in orally exhaled gas was 8.7 parts per billion (ppb) (95% confidence interval (C.I.), 8.1-9.2 ppb) and sampled data followed a log-normal distribution (Kolmogorov-Smirnov d = 0.77, P0.2). No difference was found between boys (mean value, 8.4 ppb; 95% C.I., 7.3-9.4 ppb) and girls (mean value, 8.9 ppb; 95% C.I., 7.9-9.9 ppb). No significant correlation was found between age, height, or spirometric data and exhaled NO levels (r0.2). The mean value of nasal NO concentrations was 216 ppb (95% C.I., 204-228 ppb). There was no correlation between exhaled and nasal NO values (r = 0.16, P = ns). In conclusion, this study establishes a reference range for exhaled NO values measured by a tidal breathing method in children between age 6-15 years. The observed levels are independent of age, gender, and lung function, and can be used to monitor airway inflammation in asthmatic children.

While it is known that exhaled nitric oxide (ENO) is increased in adults and school children with asthma exacerbation probably as an expression of disease activity, no studies have investigated whether this phenomenon also occurs in infants and young children with recurrent wheeze exacerbation. We measured ENO in 13 young children (mean age 20.2 mo) with recurrent wheeze (Group 1) during an acute episode and after 5 d of oral prednisone therapy. ENO was measured also in nine healthy control subjects (Group 2) (mean age 16.9 mo) and in six children with a first-time viral wheezy episode (Group 3) (mean age 11 mo). To measure ENO, infants inhaled NO-free air via a face mask from a reservoir and, through a nonrebreathing valve, exhaled in a collecting bag that was analyzed by chemiluminescence. To address the question of whether the levels of ENO collected in the bag are a reflection of the pulmonary airway, ENO determinations were performed in two healthy infants before and after tracheal intubation for elective surgery. During the acute episode of wheezing the mean (+/- SEM) value of ENO in children with recurrent wheeze (Group 1) was 14.1 +/- 1.8 ppb, almost threefold higher than in healthy control subjects (5.6 +/- 0.5 ppb, p0.001). After steroid therapy we found a mean fall of 52% in ENO (5.9 +/- 0.7 ppb, p0.01) compared with baseline values. ENO values measured before and after intubation in two infants were 6 ppb and 5 ppb in one child and 7 ppb and 6 ppb in the other one. The mean value of ENO of children with first-time wheeze (Group 3) was 8.3 +/- 1.3 ppb, significantly lower (p0.05) than the value of children with recurrent wheeze (Group 1). In conclusion, we describe a method to measure ENO in young children and show that infants with recurrent wheeze have elevated levels of ENO during exacerbation that rapidly decrease after steroid therapy. This suggests that, in these children, airway inflammation could be present at a very early stage.

Nitric oxide (NO) produced in human airways seems to have both homeostatic and proinflammatory actions in the respiratory system. NO production has been shown to be higher in the exhaled air of asthmatic adults than in normal subjects. The aim of this study was to evaluate exhaled NO production during asthma exacerbation in children and the effect of a rescue course of oral steroid therapy.We measured NO in the exhaled air of 16 children (8 girls and 8 boys, aged 6 to 13 years) with an acute asthmatic episode before and after 5 days of therapy with prednisone, and in 16 healthy children. To measure NO, children inhaled NO-free air and, breathing at tidal volume, exhaled in a circuit from which a chemiluminescence analyzer sampled continuously. To assess the effect of acute changes in bronchial caliber on exhaled NO levels, we measured NO before and after a positive bronchodilation test result with albuterol in seven children with asthma whose disease was stable.In the group with acute asthma (forced expiratory volume in 1 second 62% +/- 4.4% predicted, mean +/- SEM), NO levels were significantly higher (31.3 +/- 4.2 parts per billion [ppb]) than in healthy children (5.4 +/- 0.4 ppb, p0.001). Administration of prednisone (1 mg/kg per day orally) for 5 days resulted in a mean decrease of 46% +/- 4% in exhaled NO concentrations (16.5 +/- 2.3 ppb, p0.001) compared with baseline, accompanied by a significant improvement in lung function (forced expiratory volume in 1 second 90.7% +/- 4.3% predicted). However, in patients with asthma exhaled NO levels remained significantly higher than in control children (p0.001) after steroid treatment. When exhaled NO was measured before and after a positive result after bronchodilator reversibility testing, we found no difference in exhaled NO levels (24 +/- 3.8 ppb vs 23.8 +/- 3 ppb; difference not significant). This demonstrates that inhaled albuterol and acute changes in bronchial caliber do not affect exhaled NO measurement.These data show that children with asthma exacerbation have high levels of exhaled NO that rapidly decrease with oral steroid therapy. We suggest that measurement of exhaled NO may represent a noninvasive method of monitoring airway inflammation in children with asthma.

* Abbreviations: HPS = : hepatopulmonary syndrome • NO = : nitric oxide • ppb = : parts per billion The hepatopulmonary syndrome (HPS) has been defined as the triad of liver disease, arterial hypoxemia, and intrapulmonary vascular dilatation that causes right-to-left shunting of blood flow.1 ,2 As in adults, HPS in children also occurs in many types of liver diseases and the prevalence of this complication may range from 5% to 29%.3 The clinical findings of the syndrome include varying signs and symptoms of liver disease, finger clubbing, shortness of breath, cyanosis, dyspnea, platypnea, and orthodeoxia. Arterial hypoxemia may be so severe (Pao 2 = 50 mm Hg while breathing room air) that continuous oxygen supplementation is required and is associated with poor outcome.2 ,4 Several reports suggest that orthotopic liver transplantation can lead to complete resolution of severe hypoxemia.3 ,5 ,6 Adult patients with HPS have been reported to have significantly higher exhaled nitric oxide (NO) levels compared with healthy controls and with cirrhotics without hypoxemia.7 ,8 The mechanisms that lead to an increased production of exhaled NO are still unknown but this vasoactive gas seems to be involved in the pathogenesis of the hemodynamic abnormalities of this syndrome. To our knowledge this phenomenon has not been observed in children. The purpose of this report was to evaluate exhaled NO levels in a 6-year-old boy affected by HPS. A 6-year-old boy was referred to our hospital for the evaluation of liver transplantation attributable to HPS. At the age of 1 year the patient had growth, speech, and motor development delay. At 30 months he presented with altered liver function tests (serum glutamic–oxaloacetic transaminase 84 U/L, alkaline phosphatase 872 U/L, … Address correspondence to Eugenio Baraldi, MD, Department of Pediatrics, University of Padova, Via Giustiniani 3, 35128 Padova, Italy. E-mail: eugi{at}child.pedi.unipd.it