Reproductive isolation of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) in southern Spain

The phenology, reproductive cycle and cone crop of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) were studied during three consecutive years in southern Spain. The flowering pattern of each subspecies was constant during this period: inJ. phoenicea subsp.turbinata flowering was always in October–November, whileJ. phoenicea subsp.phoenicea flowered in February–March. There was no overlap between the flowering periods of the two taxa. The reproductive cycle of the two subspecies was similar, though there were some important differences because of their flowering times. Male cone development in both subspecies occurred in autumn, but the male cones ofJ. phoenicea subsp.phoenicea did not shed pollen until the end of February. InJ. phoenicea subsp.turbinata, the gap between pollination and fertilisation was seven months while inJ. phoenicea subsp.phoenicea it was only three months, fertilisation taking place at the same time in the two taxa. Post-fertilisation events were similar in the two subspecies, leading to a cycle of two years inJ. phoenicea subsp.turbinata and a shorter one of about 20 months inJ. phoenicea subsp.phoenicea. Alternating good and slight seed cone crops were found in both taxa during the three years studied, the good crops of one subspecies coinciding with the slight crops of the other one. Although the proximity of the two subspecies and method of pollination could make hybridisation between the taxa possible, the different flowering seasons prevent hybridisation isolating them reproductively.     

Aeropalynological and phenological study in two different Mediterranean olive areas: Cordoba (Spain) and Perugia (Italy)

ABSTRACT

An aerobiological and phenological investigation on the olive tree was carried out during three years in two areas: Cordoba (Spain) and Perugia (Italy). In these countries, this species is economically important and those areas were chosen because of the long series of aeropalynological data (1982–1998) available, obtained by means of identical volumetric pollen traps. The aim of this study was to use phenological observations to prove the real contribution to the pollen curves in different cultivated areas. Results show that in Cordoba province (302.152 ha) the pollen curve is characterised by different peaks because of the pollination of different cultivated crops. In some cases, these crops are located far from the pollen trap (50 km) but pollen is transported thanks to favourable winds during the flowering period. In Perugia (750 ha) the pollen curve is characterised by only one peak; it is very concentrated because of the proximity of the investigated crops. The objective of this research was to obtain information on this species in order to elaborate statistical models aimed at forecasting the potential fruit production based on the amount of pollen released into the atmosphere.     

Phenological olive chilling requirements in Umbria (Italy) and Andalusia (Spain)

Phenological observations in olive tree (Olea europaea L.), on the first reproductive phase (budburst) and the winter chilling temperatures required for its onset, were analysed over a 4-year period (1998 - 2001). Research was carried out on two different cultivars growing in two Mediterranean olive-growing areas: 'Ascolana' in central Italy and 'Picudo' in southern Spain. The two main objectives of the study were: (1) to evaluate the different amounts of winter chilling, and their relationship with the budburst dates in outdoor olive plantations; and (2) to test the validity in the study areas of two chilling calculation methods, the Aron and the Utah method. Results show that for the Spanish cultivar the average chilling requirement of 997 h was approximately half of that recorded for the Italian cultivar (1848 h). Also, the year-to-year variability in chilling accumulation and the reproductive development date was observed to be higher in the Spanish area than in the Italian one, indicating a more consistent and predictable winter chilling response in the latter. As regards the validity of the methods, the Aron method seems to be more appropriate for use in warmer places, since it yielded better results in the Spanish site.     

Comparison of Alternaria spore levels between two areas within the same city (Salamanca,Middle West Spain)

Aerobiologia - The purpose of this study is to contribute to the knowledge about fungal spores in the atmosphere of the city of Salamanca (Middle West Spain), through the comparative study of...     

Comparison of methods used in zooplankton sampling and counting in the joint Russian-Finnish evaluation of the trophic state of Lake Ladoga

The results of a joint Russian-Finnish investigation on zooplankton in Lake Ladoga are presented, and a comparison is made between two sampling techniques, tube sampler and plankton net, and between two counting methods. The precision of subsampling and sampling is further discussed on the basis of zooplankton data gathered in Lake Saimaa, Finland. The comparison clearly indicates that a tube sampler is required for reliable sampling of small-sized animals, while a plankton net saves time and is a more economical sampler for large, rare or active animals. The comparison between the results obtained by Finnish and Russian workers, using different counting procedures, shows that the main groups of crustacean zooplankton are similarly counted and identified in the two laboratories.     

Nosema ceranae infection in honeybee samples from Tuscanian Archipelago (Central Italy) investigated by two qPCR methods

Nosema apis and Nosema ceranae are microsporidian parasite worldwide spread causing an emerging infectious disease of European honeybee Apis mellifera. The Nosema presence was deeply investigated in several countries but low information are presents about islands. In this investigation was evaluated the presence N. ceranae and N. apis in apiaries located in Tuscanian Archipelago islands (Central Italy). For N. ceranae detection, two different Real-Time PCR (qPCR) methods, the 16S rRNA and Hsp70 gene amplification qPCR, were performed on honey bee samples; while, for N. apis only the 16S rRNA qPCR amplification was performed. On all islands, only N. ceranae was present, while N. apis was not found in the samples. The two qPCR showed significant difference (p < 0.040) in N. ceranae spores quantification. The single-copy Hsp70 gene method qPCR assay systematically detected a lower amount of N. ceranae copies compared to the multi-copy 16S rRNA gene method.     

Reproductive biology ofFrangula alnus (Rhamnaceae) in southern Spain

Various aspects of the reproductive biology of the perennial hermaphroditeFrangula alnus Mill. were studied in two populations located in the province of Cádiz, southern Spain. Flowering extends from the second week of May to early July. The small, whitegreenish, entomophilous flowers are incompletely protandrous and last 8–10 days, but pollen transfer takes place only on days 1–3. A very weak, pleasant odour along with nectar and pollen attract over 45 insect species, of which 21 are probable pollinators. These are mainlyDiptera but also includeHymenoptera and the unusual (as flower visitors)Mecoptera. Reproduction is exclusively sexual and strictly xenogamous, with pollen transfer depending solely on insect vectors. Although flower morphology and individual flower phenology do not fully prevent self-pollination, and geitonogamy can easily take place, the level of autogamy was negligible. Therefore, some self-incompatibility mechanism is operative in this species. Only 2.8% of open-pollinated flowers set fruit. At the flower level, fruit initiation was apparently limited by availability of cross-pollen, which in turn seemed influenced by the structure of the population. The seed/ovule ratio in ripe fruits was ca. 50%. Predispersal (maternal) reproductive success (percent ovules becoming filled seeds) was 1.42%. For mature individuals this corresponded to ca. 430 to 1560 potential offspring per year.     

Hybrid zones between two genetically differentiated forms of the pond frog Rana lessonae in southern Italy

Two distinct population groups of the pond frog Rana lessonae were detected in peninsular Italy and Sicily by multilocus electrophoresis: one group inhabits the peninsula down to northern Calabria, the second occurs in southern Calabria and on Sicily. Fixed alternative alleles distinguish the two groups at 5 of the 25 loci examined; marked allele frequency differences were observed at two additional loci. On average, the two groups differ by a Nei's standard genetic distance of 0.4. A wide hybrid zone (about 120 km) occurs between the two groups, with high genotypic diversity and absence of pure parental genotypes in central Calabria. Patterns of allozyme variation suggest that at least two distinct contact and hybridization events occurred, one in the Catanzaro, the other in the Crati-Sibari plains, about 70 km to the north. Geological evidence indicates that these areas correspond to two main marine-flooded grabens that would have repeatedly interrupted or reduced genetic exchange during Plio-Pleistocene times. The finding of a fixed difference at the Mdhp-1 locus between Sicilian and Calabrian R. lessonae witnesses their continuing differentiation following their last separation by definitive opening of the Strait of Messina, about 50 000 years ago. The wide hybrid zone, the diversity of genotypes and the agreement with Hardy-Weinberg expectations suggest complete hybrid fertility. Different patterns of introgression were observed at the various loci. The pattern of allelic variation at loci in R. lessonae is paralleled by the pattern of variation in lessonae genomes of the sympatric hemiclonal hybrid Rana esculenta, into which lessonae genomes are introduced each generation as a result of hybridogenesis.     

Comparison of the checkerboard and E-test methods used for the analysis of two antibiotics combination

The aim of this study was to compare checkerboard method with E-test assay for interaction analysis of aminoglycosides in combination with other antibiotics on selected clinical bacterial strains. In the first step, MIC values of analysed antibiotics, against particular bacterial strain were established. In the next step, antibiotics interaction was analysed by checkerboard technique and E-test. We found some difficulties while comparing these two methods. The checkerboard and E-test results corresponded in about 55%. Twenty-one percentage of results obtained by both methods showed some discrepancies. In 15% of cases, because of high MICs values, comparison of the results was impossible. Some investigators declare FIC indexes, from over 0.5 up to 4, for neutral effects. Sharing this point of view, above 21% of discrepancies results agreed. In such situation, definite disagreement was observed only in 8% of obtained results. In this investigation, additive and neutral effects were dominant. The E-test technique is less-laborous than standard agar method. In this study the E-test assay indicated synergy in only one case. Because of the manner in which the E-test strips were placed on the agar (scales intersecting at the MICs) only dramatic cases of antagonism were detected. Mild cases were undetected because the inhibition zone run under the crossed strips and was therefore unreadable and interpreted as indifference. On the basis of these results, examination of interactions between antibiotics by E-test, appears to be possible alternative to checkerboard method with mentioned limitations.     

Potential pathogens carried by Spanish ibex (Capra pyrenaica hispanica) in southern Spain

The Spanish ibex (Capra pyrenaica hispanica) population of southern Spain was surveyed for potential pathogens associated with the conjunctiva, external ear canal, as well as reproductive and upper respiratory tracts. We sampled 321 ibex (131 adult males, 100 adult females, and 90 yearlings); these included 271 apparently healthy animals and 50 that were naturally infected with Sarcoptes scabiei. A total of 688 bacterial isolates were identified (377 gram-negatives, 225 gram-positives, and 86 Mycoplasma spp.); sex, age, location, infection with S. scabiei, and disposition of the animal (free-ranging versus captive) were evaluated as risk factors for infection. Infections with Mycoplasma agalactiae and Mycoplasma arginini were associated with age, having a higher frequency of isolation in young animals. With Escherichia coli, Mannheimia haemolytica, Pasteurella multocida biotype A, and Staphylococcus aureus, significantly higher isolation rates were associated with adults. The isolation frequency for E. coli was higher in females, whereas Moraxella bovis isolations were mostly associated with males. The presence of mange increased the risk of infection with both Streptococcus equi subsp. zooepidemicus and M. haemolytica. The geographic origin of sampled animals was related to the isolation of Branhamella ovis, M. agalactiae, and all Pasteurella sp. Isolations of M. haemolytica, P. multocida biotype A, E. coli, and B. ovis were more prevalent in samples from free-ranging rather than captive animals. Of the gram-positive bacteria, S. aureus represented the predominant species isolated from nasal, vaginal, and ocular samples. Mycoplasma agalactiae and M. arginini were the predominant Mycoplasma spp., and both were associated most often with the external ear canal. The most frequently isolated gram-negative bacteria included E. coli, M. haemolytica, P. multocida biotype A, and B. ovis. Isolation rates of gram-negative species varied by source. In nasal samples, M. haemolytica and P. multocida biotype A were isolated most frequently, whereas in ocular and vaginal samples, B. ovis and E. coli, respectively, were most frequently isolated.     

Thelazia gulosa Railliet & Henry, 1910 and T. skrjabini Erschow, 1928 infection in southern Europe (Italy)

The occurrence of Thelazia gulosa and T. skrjabini in cattle in Italy, together with an update of the prevalence of Thelazia spp. in the southern Italy, is reported. On 764 slaughtered native cattle, fifty-five (7.2%) bovines were infected by Thelazia spp: T. rhodesi was found in 44 (80%) animals, T. gulosa was found in 19 (34.5%) cattle and only one (1.8%) cattle harboured T. skrjabini. This is the first report of T. gulosa and T. skrjabini in Southern Europe.     

Comparison between tracheal ratio methods used by three observers at three occasions in English Bulldogs

Background

Tracheal hypoplasia is a congenital condition described in mainly brachycephalic breeds and is one component of the brachycephalic obstructive airway syndrome (BOAS). Two radiographic methods have been described to evaluate the dimensions of the tracheal diameter in dogs and to distinguish between hypoplastic and non-hypoplastic tracheas: the tracheal lumen diameter to thoracic inlet distance ratio (TD/TI) and the ratio between the thoracic tracheal luminal diameter and the width of the proximal third of the third rib (TT/3R). The purpose of this study was to compare these two published radiographic methods between observers, different measuring occasions and to investigate the effect on classification of dogs as having hypoplastic or non-hypoplastic tracheas using four previously published mean ratios as cut-offs (<0.11, <0.127 and <0.144 for the TD/TI and <2.0 for the TT/3R method).Three observers evaluated right and left lateral recumbent radiographs from 56 adult English Bulldogs independently on three different occasions. TD/TI and TT/3R ratios were calculated and correlated between measuring occasions. Kappa, observed, positive, and negative agreements were calculated between observers and measuring occasions. Number of hypoplastic and non-hypoplastic dogs for each method and occasion was determined using <0.11, <0.127 and <0.144 as cut-offs for TD/TI and <2.0 for TT/3R.

Results

Intraobserver agreement varied with kappa between 0.45-0.94 for the TD/TI and 0.20-0.86 for the TT/3R method. Interobserver kappa varied between 0.27-0.70 for the TD/TI method and between 0.05-0.57 for the TT/3R method. There was poor agreement in classifying English Bulldogs as tracheal hypoplastic or non-hypoplastic, depending on measuring method, cut-off value and observer.

Conclusions

The diagnostic value of both the TD/TI and TT/3R methods with such poor agreement is questionable, and significantly impacts their reliability for both clinical evaluation of dogs and use in health screening programs.

     

Inequality in mortality in pre-industrial southern Europe during an epidemic episode: socio-economic determinants (eighteenth - nineteenth centuries Spain)

The objective of this study is to gain more comprehensive knowledge about social inequality in mortality in pre-industrial periods. With this aim, we have reconstructed the life courses of the inhabitants of the town of Vera in south-east Spain for the period 1797–1812 in order to estimate the influence of socio-economic status on ordinary and extraordinary mortality, given that, during this period, the town suffered from several epidemic outbreaks of yellow fever. As a result of these outbreaks, around a quarter of the town’s population died. The results obtained indicate social inequality in mortality at least from the end of the eighteenth century. Although the differences are higher in mortality caused by non-infectious diseases or ill-defined causes, the coefficients also show a certain social gradient in mortality derived from infectious diseases. However, with respect to this latter type of mortality, the place of residence - seems to have a greater influence on the chances of survival than socio-economic status.     

两种粪便保存方法对肠道菌群结构研究的影响

目的比较2种粪便保存方法（室温法和Invitek公司的粪便稳定剂保存法）对菌群结构研究的影响。方法应用PCR-变性梯度凝胶电泳技术（PCR-DGGE）方法,对用2种方法保存的3位志愿者粪便样品进行菌群结构的分析。结果室温法保存粪便样品24h后,S1个体菌群结构与原始样品的菌群结构相似度为83％,S2和S3的菌群结构与其原始样品的相似度仅为77％。而使用粪便稳定剂保存1d,期间各时间点样品菌群结构与原始样品相比变化较小,相似度在80％-90％。结论粪便稳定剂具有一定的稳定样品菌群结构的作用,在新鲜粪佰样品不能寺刻讲行深冻的情况下,使用粪便稳定剂是一种较好的样品保存方法。     

Bioclimatic analysis in a region of southern Italy (Calabria)

In this study, an analysis of precipitation and temperature data has been performed over 67 series observed in a region of southern Italy (Calabria). At first, to detect possible trends in the time series, an analysis was performed with the Mann–Kendall non-parametric test applied at monthly and seasonal scale. An additional investigation, useful for checking the climate change effects on vegetation, has also been included analysing bioclimatic indicators. In particular, Emberger, Rivas-Martinez and De Martonne indices were calculated by using monthly temperature and precipitation data in the period 1916–2010. The spatial pattern of the indices has been evaluated and, in order to link the vegetation and the indices, different indices maps have been intersected with the land cover data, given by the Corine Land Cover map. Moreover, the temporal evolution of the indices and of the vegetation has been analysed. Results suggest that climate change may be responsible for the forest cover change, but, given also the good relationship between the various types of bioclimate and forest formations, human activities must be considered.     

Olive flowering trends in a large Mediterranean area (Italy and Spain)

The aim of this study was to investigate the main climatic and biological trends related to olive flowering in central-southern Italy compared to those in Andalusia, Spain. Results since 1982 were compared for the two long-series monitoring areas of Cordoba and Perugia, and since 1992–1999 for the short-series areas. The relationship between climatic trends and the biological response of the olive, a widespread culture in the Mediterranean basin, were investigated. An aerobiological method involving capturing pollen released into the atmosphere was utilised as a bioindicator of flowering phenology. The study results confirm the strong relationship between flowering periods and spring temperature trends for the olive. Temperature during March, April and May was the parameter most related to flowering date in the study areas, particularly in Italy. In some cases we found a significant correlation between flowering and past autumn temperatures, probably due to their effect on floral bud dormancy induction, but this phenomenon appeared to be of minor importance in the studied areas. The phenological trend results show the continuous advance of flowering dates to the late 1990s, followed by a relatively stationary time series related to a short-term temperature fluctuation in the Mediterranean area. This latter period probably represents a mesoscale event forced by a macroscale event—the North Atlantic Oscillation. The results reveal that the trend towards increased temperatures, and the consequent flowering advance of some species, indicated some years ago is nowadays not as clear as was expected and should be confirmed over the next few years in the Mediterranean areas under investigation.