The influence of meteorological parameters on <Emphasis Type="Italic">Alternaria</Emphasis> and <Emphasis Type="Italic">Cladosporium</Emphasis> fungal spore concentrations in Beja (Southern Portugal): preliminary results

Introduction Fungal spores constitute an important fraction of bioaerosols in the atmosphere. Objectives To analyse the content of Alternaria and Cladosporium spores in the atmosphere of Beja and the effect of meteorological conditions on their concentrations. Methodology The daily and hourly data of Alternaria and Cladosporium fungal spores concentration in the atmosphere of Beja were monitored from April 12, 2012 to July 30, 2014, based on the Portuguese Aerobiology Network methodology. The influence of meteorological conditions on the studied types of fungal spore concentrations was assessed through Spearman’s correlation analysis. Results During the study period, 20,741 Alternaria spores and 320,862 Cladosporium spores were counted. In 2013, there were 5,822 Alternaria spores and 123,864 Cladosporium spores. The absolute maximum concentrations of Alternaria and Cladosporium spores were recorded on November 8, 2013, with 211 and 1301 spores/m³, respectively. Temperature, insolation and wind direction parameters showed a positive correlation with Alternaria and Cladosporium spore levels, while relative humidity and precipitation presented a negative correlation, which is statistically significant. Wind speed only showed a statistically significant positive correlation in terms of Alternaria spore levels. Conclusion Alternaria and Cladosporium spores are present in the atmospheric air of Beja throughout the year, with the highest concentration period occurring during spring and autumn. There was a clear effect of meteorological parameters on airborne concentrations of these fungal spores.     

First volumetric record of fungal spores in the atmosphere of Montevideo City,Uruguay: a 2-year survey

In Uruguay, aeromycological studies are restricted to a gravimetric analysis performed from December 1942 to March 1944 in Montevideo where spores of Pucciniaceae, Alternaria and Helminthosporium were the only specimens identified. Daily monitoring of airborne fungal spores was carried out for the first time in Montevideo, from April 2012 to March 2014, using a Rotorod sampler in order to evaluate the seasonal variation and influence of meteorological parameters. A total of 548,309.68 spores/m³ were recorded which belong to anamorphs of Higher Fungi (69.18 %), Phyla Ascomycota (12.62 %), Basidiomycota (8.01 %), Oomycota (0.37 %) and Myxomycota (0.06 %). Airborne spores occurred in Montevideo throughout the whole year. However, a seasonal pattern was revealed, with the highest concentrations recorded in autumn and summer. The most abundant spore types were Cladosporium (53.22 %), Alternaria (6.62 %), Didymella Group (5.86 %), Leptosphaeria Group (4.37 %) and Coprinus (4.3 %). Temperature appeared to be the most influential meteorological factor correlating significantly and positively with total spore, Cladosporium, Alternaria and Didymella Group abundance. Relative humidity influenced positively total spore, Cladosporium and Didymella Group concentrations while a weak negative association was obtained for Alternaria. Wind speed correlated negatively with total spore, Cladosporium, Alternaria and Didymella Group. Precipitation showed a negative influence on Alternaria, while positive correlations were observed for Didymella Group. For the first time, fungal spores considered allergenic were recorded in Montevideo atmosphere and the risk of exposure would have been high from December to June. However, long-term sampling is needed to define seasonal prevalence patterns and the influence of meteorological conditions on spore abundance.     

Phenotypic and genotypic diversity of airborne fungal spores in Demänovská Ice Cave (Low Tatras,Slovakia)

This paper is the first aero-mycological report from Demänovská Ice Cave. Fungal spores were sampled from the internal and external air of the cave in June, 2014, using the impact method with a microbiological air sampler. Airborne fungi cultured on PDA medium were identified using a combination of classical phenotypic and molecular methods. Altogether, the presence of 18 different fungal spores, belonging to 3 phyla, 9 orders and 14 genera, was detected in the air of the cave. All of them were isolated from the indoor samples, and only 9 were obtained from the outdoor samples. Overall, airborne fungal spores belonging to the genus Cladosporium dominated in this study. However, the spores of Trametes hirsuta were most commonly found in the indoor air samples of the cave and the spores of C. herbarum in the outdoor air samples. On the other hand, the spores of Alternaria abundans, Arthrinium kogelbergense, Cryptococcus curvatus, Discosia sp., Fomes fomentarius, Microdochium seminicola and T. hirsuta were discovered for the first time in the air of natural and artificial underground sites. The external air of the cave contains more culturable airborne fungal spores (755 colony-forming units (CFU) per 1 m³ of air) than the internal air (from 47 to 273 CFU in 1 m³), and these levels of airborne spore concentration do not pose a threat to the health of tourists. Probably, the specific microclimate in the cave, including the constant presence of ice caps and low temperature, as well as the location and surrounding environment, contributes to the unique species composition of aeromycota and their spores in the cave. Thus, aero-mycological monitoring of underground sites seems to be very important for their ecosystems, and it may help reduce the risk of fungal infections in humans and other mammals that may arise in particular due to climate change.     

Molecular characterization and expression analysis of the Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> exchanger gene family in <Emphasis Type="Italic">Medicago truncatula</Emphasis>

One important mechanism plants use to cope with salinity is keeping the cytosolic Na⁺ concentration low by sequestering Na⁺ in vacuoles, a process facilitated by Na⁺/H⁺ exchangers (NHX). There are eight NHX genes (NHX1 through NHX8) identified and characterized in Arabidopsis thaliana. Bioinformatics analyses of the known Arabidopsis genes enabled us to identify six Medicago truncatula NHX genes (MtNHX1, MtNHX2, MtNHX3, MtNHX4, MtNHX6, and MtNHX7). Twelve transmembrane domains and an amiloride binding site were conserved in five out of six MtNHX proteins. Phylogenetic analysis involving A. thaliana, Glycine max, Phaseolus vulgaris, and M. truncatula revealed that each individual MtNHX class (class I: MtNHX1 through 4; class II: MtNHX6; class III: MtNHX7) falls under a separate clade. In a salinity-stress experiment, M. truncatula exhibited ~?20% reduction in biomass. In the salinity treatment, sodium contents increased by 178 and 75% in leaves and roots, respectively, and Cl^? contents increased by 152 and 162%, respectively. Na⁺ exclusion may be responsible for the relatively smaller increase in Na⁺ concentration in roots under salt stress as compared to Cl^?. Decline in tissue K⁺ concentration under salinity was not surprising as some antiporters play an important role in transporting both Na⁺ and K ⁺ . MtNHX1, MtNHX6, and MtNHX7 display high expression in roots and leaves. MtNHX3, MtNHX6, and MtNHX7 were induced in roots under salinity stress. Expression analysis results indicate that sequestering Na⁺ into vacuoles may not be the principal component trait of the salt tolerance mechanism in M. truncatula and other component traits may be pivotal.     

Dynamics and diversity of a microbial community during the fermentation of industrialized <Emphasis Type="Italic">Qingcai</Emphasis> paocai,a traditional Chinese fermented vegetable food,as assessed by Illumina MiSeq sequencing,DGGE and qPCR assay

Paocai is a traditional Chinese fermented food and typically produced via spontaneous fermentation. We have investigated the microbial community utilized for the fermentation of industrialized Qingcai paocai using the combination of Illumina MiSeq sequencing, PCR-mediated denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative PCR (qPCR) assay. Three main phyla, namely Firmicutes, Proteobacteria and Bacteroidetes, were identified by both MiSeq sequencing and PCR-DGGE. The dominant genera observed in the fermentation were Lactobacillus, Pseudomonas, Vibrio and Halomonas. Most genera affiliated with Proteobacteria or Bacteroidetes were detected more often during the earlier part of the fermentation, while Lactobacillus (affiliated with Firmicutes) was dominant during the later fermentation stages. Fungal community analysis revealed that Debaryomyces, Pichia and Kazachstania were the main fungal genera present in industrialized Qingcai paocai, with Debaryomyces being the most dominant during the fermentation process. The quantities of dominant genera Lactobacillus and Debaryomyces were monitored using qPCR and shown to be 10⁹–10¹² and 10⁶–10¹⁰ copies/mL, respectively. During the later fermentation process of industrialized Qingcai paocai, Lactobacillus and Debaryomyces were present at 10¹¹ and 10⁸ copies/mL, respectively. These results facilitate further understanding of the unique microbial ecosystem during the fermentation of industrialized Qingcai paocai and guide future improvement of the fermentation process.     

Characterization of biofilm-forming marine bacteria and their effect on attachment and germination of algal spores

In this work, 37 bacterial strains isolated from biofouling of marine organisms and from the Museum of Heterotrophic Bacteria of the National Scientific Center of Marine Biology were studied. The strains were identified based on their phenotypic characteristics and on the fatty acid composition of their cell wall lipids. Members of the genus Pseudoalteromonas prevailed both in associated microflora of two dinoflagellate clones and in the biofilms from marine hydrobionts. Associated microflora included also members of the CFB cluster, Bacillus, Sulfitobacter, Acinetobacter, Shewanella, and Psychrobacter. A considerable portion of strains (48.6%) exhibited antimicrobial activity. Antifouling activity against algal spores was studied using single-species bacterial biofilms and the spores of Ulva lactuca и Undaria pinnatifida, the algae most common in the Sea of Japan. Strong inhibitory effect on attachment of Ulva and Undaria spores was observed for 75 and 51% of the strains, respectively. Attached spores were, however, less sensitive to the inhibitory action of biofilms. Species specificity of algal response to bacteria was shown, with a strain having different effect on the spores of different algal species. Biotechnologically promising strains were determined, which exhibited high activity against the spores of macroalgae and could probably be used as producers of antifouling substances and as components of antifouling coatings. No relation was found between antifouling activity of bacteria and the source of their isolation. Our results indicate wide occurrence of bacteria with antifouling activity among associated microflora of marine hydrobionts and demonstrate the extent of complexity and diversity of relations between bacterial biofilms and algal spores.     

Airborne fungal spore distribution in Bangkok,Thailand: correlation with meteorological variables and sensitization in allergic rhinitis patients

Information about airborne fungal spore is crucial for health risk assessment and management, especially for patients with allergy and asthma. Nonetheless, such data are rarely available from certain areas of the world, including Southeast Asia. The aim of this study was to gain updated information about airborne fungal spore in Bangkok, the capital city of Thailand. A survey was conducted at five sampling sites in Bangkok, using the Rotorod Sampler^® for a period of 1 year. High concentrations of spores were found all year with the peak between August and November. The most prominent spore types were Cladosporium, Nigrospora, Puccinia, Aspergillus/Penicillium, and Fusarium. The spore concentrations were positively and significantly correlated with the amount of rainfall and relative humidity, reaching the maximum level in September. Sensitization rates to Cladosporium, Penicillium, and Aspergillus among Thai atopic patients were approximately 16.6, 13.6, and 13.0%, respectively.     

Pollen analysis of spider web samples from Korba District,Chhattisgarh (central India): an aerobiological aspect

Pollen analyses of spider web samples, collected from the recently planted Tectona grandis (T. grandis, teak)-dominated tropical deciduous forest of Bhulsidih Village, Korba District (Chhattisgarh, central India), shed light on the relationship between the extant vegetation and pollen rain. The study revealed the dominance of pollen of herbs and trees, whereas shrubs, fern spores and algal remains are meagre. Among the tree taxa, Sapotaceae, Syzygium, Holoptelea, Lannea coromandelica, Shorea robusta and Grewia are dominating with moderate to low and intermittent presence of Madhuca indica, Terminalia, Mitragyna, Schleichera, Anacardiaceae, Diospyros, Emblica officinalis and Flacourtia. However, the rest of the forest constituents are either not represented at all despite their presence in the floristics, which could be attributed to their low pollen productivity owing to entomophily as well as their poor pollen preservation pattern. On the other hand, the ground vegetation is represented by the very high frequency of grasses (Poaceae) along with Tubuliflorae, Chenopodiaceae/Amaranthaceae and Cerealia, however, Artemisia, Xanthium, Malvaceae, Caryophyllaceae and Justicia in moderate to lower values. Ferns, which occur abundantly along the adjoining stream banks, are marked by the sporadic retrieval of trilete spores that could be ascribed to the prevailing damp condition around the sampling provenance. The study, in addition to understanding the pollen–vegetation relationship, could also be helpful in aerobiological study, especially in assessing the allergenicity of various pollen grains/spores in the area of investigation, causing bronchial asthma, hay fever (allergic rhinitis/pollinosis), naso-bronchial allergy and other respiratory disorders along with conjunctivitis, contact dermatitis, eczema, food allergies and other health disorders.     

The survival of micromycetes exposed to space conditions

Original data on the survival of fungal spores exposed to space conditions are presented. The experiment was carried out on the Earth-orbiting Russian satellite Foton-M4. The flight duration of the satellite was 45 days. Thirteen fungal species (hyaline as well as pigmented) from 10 genera recovered from destructed stone materials were studied. Sterile quartz sand was inoculated by the fungal spores and was placed into Eppendorf tubes. During the space flight, the Eppendorf tubes with fungal spores were kept inside the Foton descent capsule in the “Biokont” containers and on the external surface of the capsule in the “Exobiofrost” containers exposed to the open space as well. Spores of ten species (77% of all tested species), i.e. Acremonium charticola, Aspergillus niger, Aspergillus versicolor, Chaetomium globosum, Cladosporium sphaerospermum, Penicillium chrysogenum, Penicillium verrucosum, Purpureocillium lilacinum, Sarocladium kiliense, and Trichoderma harzianum, survived after the flight both inside and outside the descent capsule. Only three species (23% of all tested species), i.e. Acremonium furcatum, Engyodontium album and Verticillium zaregamsianum, failed to survive outside as well as inside the capsule. Spore viability differed depending on the fungal species. Thus, spores of some fungal species are able to survive under the complex of stress factors such as low temperature values, radiation, etc. We have shown that micromycetes can be used as a model group for study of eukaryotic organisms’ resistance to stress factors, due to their high tolerance not only to extreme terrestrial environments, but to the extraterrestrial ones as well.     

Survey of Philippine coffee beans for the presence of ochratoxigenic fungi

In 2012 to 2014, Philippine green coffee beans from Coffea arabica in Benguet and Ifugao; Coffea canephora var. Robusta in Abra, Cavite, and Ifugao; and Coffea liberica and Coffea excelsea from Cavite were collected and assessed for the distribution of fungi with the potential to produce ochratoxin A (OTA). The presence of fungal species was evaluated both before and after surface sterilization. There were remarkable ecological and varietal differences in the population of OTA-producing species from the five provinces. Aspergillus ochraceus, A. westerdijkiae, and Penicillium verruculosum were detected from Arabica in Benguet and Ifugao while Aspergillus carbonarius, Aspergillus niger, and Aspergillus japonicus were isolated in Excelsa, Liberica, and Robusta varieties from Abra, Cavite, and Davao. Contamination by Aspergillus and Penicillium species was found on 59 and 19 %, respectively, of the 57 samples from five provinces. After disinfection with 1 % sodium hypochlorite, the levels of infection by Aspergillus and Penicillium fell to 40 and 17 %, respectively. A total of 1184 fungal isolates were identified to species level comprising Aspergillus sections Circumdati (four species), Clavati (one), Flavi (one), Fumigati (one), Nigri (three), and Terrie (one). Within section Circumdati, 70 % of A. ochraceus produced OTA as high as 16238 ng g^?1 while 40 % of A. westerdijkiae produced maximum OTA of 36561 ng g^?1 in solid agar. Within section Nigri, 16.76 % of A. niger produced OTA at the highest 18439 ng g^?1, 10 % of A. japonicus at maximum level of 174 ng g^?1, and 21.21 % of A. carbonarius yielded maximum OTA of 1900 ng g^?1. Of the 12 species of Penicillium isolated, P. verruculosum was ochratoxigenic, with a maximum OTA production of 12 ng g^?1.     

Atmospheric concentrations and intradiurnal pattern of <Emphasis Type="Italic">Alternaria</Emphasis> and <Emphasis Type="Italic">Cladosporium</Emphasis> conidia in Tétouan (NW of Morocco)

Fungal spores of Alternaria and Cladosporium are ubiquitous components of both indoor and outdoor air samples and are the main causes of human respiratory allergies. Monitoring these airborne fungal spores during 2009–2014 was carried out by means of Hirst-type spore trap to investigate their airborne spore concentrations with respect to annual load, seasonality and overall intradiurnal pattern. Alternaria and Cladosporium spores are present throughout the year in the atmosphere of Tétouan, although they show seasonal variations. Despite important differences between years, their highest levels presented a first peak during spring and a higher second peak in summer or autumn depending on the year. The spore concentrations were homogeneously distributed throughout the day with slight increase of 7.6 and 3.7% on average between 12–14 and 14–16 h for Alternaria and Cladosporium, respectively. The borderline of 3000 sp/m³ of Cladosporium linked to the occurrence of allergic diseases was exceeded between 13 and 31 days. Airborne spores of Alternaria overcame the threshold value of 100 sp/m³ up to 95 days, suggesting that Cladosporium and Alternaria could be clinically significant aeroallergens for atopic patients.     

Aseptic rearing procedure for the stinkbug <Emphasis Type="Italic">Plautia stali</Emphasis> (Hemiptera: Pentatomidae) by sterilizing food-derived bacterial contaminants

The stinkbug Plautia stali Scott is a notorious agricultural pest whose posterior midgut hosts specific bacteria essential for its growth and survival, highlighted as an experimental model for symbiosis studies. Some symbiotic bacteria of P. stali are cultivable, found free-living in and acquired from the environment, and, furthermore, some free-living environmental bacteria are potentially capable of establishing symbiotic association with P. stali. In this context, it is expected that such environmental bacteria may occasionally contaminate and infect the experimental insects maintained in the laboratory, which could potentially affect the functional analyses of the symbiosis. Here we report that such contamination events do occur under a laboratory rearing conditions for P. stali. When symbiont-deprived newborn nymphs from surface-sterilized eggs were reared in sterilized plastic containers with autoclaved water, most of them died as nymphs presumably as a result of aposymbiosis, but only a small fraction could attain adulthood and the adult insects were all infected with γ-proteobacteria allied to Pantoea and Enterobacter. A variety of bacteria, mainly Bacillus and also Pantoea and Enterobacter, were detected from peanuts and soybeans provided as food for P. stali. Autoclaving of peanuts and soybeans eradicated these bacteria but negatively affected the host survival, whereas ethanol sterilization of peanuts and soybeans removed Pantoea and Enterobacter, but not Bacillus, without negative effects on the host survival. On the basis of these results, we established a practical procedure for aseptic rearing of P. stali, which will enable reliable and strict analyses of host–symbiont interactions in the model symbiotic system.     

Toxin Gene Contents and Activity of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Strains Against Two Sugarcane Borer Species, <Emphasis Type="Italic">Diatraea saccharalis</Emphasis> (F.) and <Emphasis Type="Italic">D. flavipennella</Emphasis> (Box)

Bacillus thuringiensis (Berliner) bears essential characteristics in the control of insect pests, such as its unique mode of action, which confers specificity and selectivity. This study assessed cry gene contents from Bt strains and their entomotoxicity against Diatraea saccharalis (F.) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). Bioassays with Bt strains were performed against neonates to evaluate their lethal and sublethal activities and were further analyzed by PCR, using primers to identify toxin genes. For D. saccharalis and D. flavipennella, 16 and 18 strains showed over 30% larval mortality in the 7th day, respectively. The LC₅₀ values of strains for D. saccharalis varied from 0.08 × 10⁵ (LIIT-0105) to 4104 × 10⁵ (LIIT-2707) spores + crystals mL^?1. For D. flavipennella, the LC₅₀ values of strains varied from 0.40 × 10⁵ (LIIT-2707) to 542 × 10⁵ (LIIT-2109) spores + crystals mL^?1. For the LIIT-0105 strain, which was the most toxic to D. saccharalis, the genes cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, cry8, and cry9C were detected, whereas for the strain LIIT-2707, which was the most toxic to D. flavipennella, detected genes were cry1Aa, cry1Ab, cry1Ac, cry1B, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, and cry9. The toxicity data and toxin gene content in these strains of Bt suggest a great variability of activity with potential to be used in the development of novel biopesticides or as source of resistance genes that can be expressed in plants to control pests.     

Successful disinfection of femoral head bone graft using high hydrostatic pressure

The current standard for sterilization of potentially infected bone graft by gamma irradiation and thermal or chemical inactivation potentially deteriorates the biomechanical properties of the graft. We performed an in vitro experiment to evaluate the use of high hydrostatic pressure (HHP); which is widely used as a disinfection process in the food processing industry, to sterilize bone grafts. Four femoral heads were divided into five parts each, of which 16 were contaminated (in duplicate) with 10⁵–10⁷ CFU/ml of Staphylococcus epidermidis, Bacillus cereus, or Pseudomonas aeruginosa or Candida albicans, respectively. Of each duplicate, one sample was untreated and stored similarly as the treated sample. The remaining four parts were included as sterile control and non-infected control. The 16 parts underwent HHP at the high-pressure value of 600 MPa. After HHP, serial dilutions were made and cultured on selective media and into enrichment media to recover low amounts of microorganism and spores. Three additional complete femoral heads were treated with 0, 300 and 600 MPa HHP respectively for histological evaluation. None of the negative-control bone fragments contained microorganisms. The measured colony counts in the positive-control samples correlated excellent with the expected colony count. None of the HHP treated bone fragments grew on culture plates or enrichment media. Histological examination of three untreated femoral heads showed that the bone structure remained unchanged after HHP. Sterilizing bone grafts by high hydrostatic pressure was successful and is a promising technique with the possible advantage of retaining biomechanical properties of bone tissue.     

Effect of cumulative polymery of tomato keeping life genes

Results from works involving the study of hetero-and homozygous interaction of the keeping quality genes alc, nor, and rin in tomato plants are presented. It is shown that cumulative polymery leading to the formation of a new “long ripening” phenotype is observed in the double heterozygotes alc/alc ⁺//nor/nor ⁺, nor/nor ⁺//rin/rin ⁺, and alc/alc ⁺//rin/rin ⁺. Strong inhibition of processes of ripeness and synthesis of carotenoids occurs with nonallelic interaction in the double homozygotes alc/alc//rin/rin, nor/nor//rin/rin, and alc/alc//nor/nor. This promotes the formation of a new “non-ripening” phenotype with the absence of visual signs of ripeness, i.e., a whitish-green coloring of the fruit.     

Substantial,and significant,expansion of ant hosts range for <Emphasis Type="Italic">Myrmicinosporidium</Emphasis> Hölldobler, 1933 (Fungi)

Six new genera, three tribes and one subfamily are added to the list of known hosts for the enigmatic endoparasitic fungus Myrmicinosporidium Hölldobler. Aphaenogaster senilis, Cataglyphis hispanica, Crematogaster auberti, Goniomma hispanicum, Messor barbarus, Tapinoma nigerrimum and Tapinoma simrothi were collected from olive groves and detected as infected with spores of the fungus. Pheidole pallidula and Tetramorium semilaeve were also found to be infected. The finding of seven hosts (seven genera, three subfamilies) from a single olive grove is an evidence that the fungus has a phylogenetically wide host spectrum and is, therefore, a generalist microparasite. Portugal is also a new country for Myrmicinosporidium.     

Intestinal enterobacteria of the hibernating <Emphasis Type="Italic">Apis mellifera mellifera</Emphasis> L. bees

Dynamics of enterobacteria of normal intestinal microflora was studied in Apis mellifera mellifera L. bees hibernating under snow in the Western Urals. The cell numbers (N) of the predominant species Klebsiella oxytoca increased from 10-10⁶ CFU/bee in November 2004 to 10⁴-10⁷ CFU/bee in March 2005; its frequency of occurrence (P) increased from 92 to 100%. Increase of Providencia rettgeri (11.2004: N up to 10⁶, P 25%; 03.2005: N 10²-10⁶, P 80%) was accompanied by the substitution of Morganella morganii (11.2004: N up to 10⁶, P 25%) with Proteus vulgaris (03.2005: N up to 10⁵, P 8%). By spring, Hafnia alvei and Citrobacter sp., which are pathogenic to bees, disappeared (11.2004: N up to 10⁵, P 13 and 10%, respectively). Endophytic species Pantoea agglomerans, Leclecria sp., and other representatives of the “Enterobacter agglomerans” group were present in November and after the first emergence in spring (N up to 10⁵; November: P 15%; April: P 23%). In April, the number of enterobacteria decreased to 10⁵, and P. rettgeri became the predominant species (P 54%) instead of K. oxytoca (P 43%).     

Atmospheric concentrations of selected allergenic fungal spores in relation to some meteorological factors,in Timişoara (Romania)

Present investigation was undertaken to study the dynamics of relationships between atmospheric fungal spores and meteorological factors in western Romania. The airborne spore sampling was carried out by employing volumetric sampling. A total of nine meteorological parameters were selected for this investigation. During 2008–2010, it was found the same pattern of behaviour in the atmosphere for selected spore types (Alternaria, Cladosporium, Pithomyces, Epicoccum and Torula). The spores occurred in the air throughout the whole year, but maximum concentrations were reached in summer. Cladosporium and Alternaria peak levels were observed in June. Epicoccum peak value was found in September. The relationships between airborne spore concentrations and environmental factors were assessed using the analysis of Spearman’s rank correlations and multiple linear regressions. Spearman’s rank correlation analysis revealed that maximum, minimum and mean temperature, and number of sunshine hours were strongly (p < 0.01) and directly proportional to the concentration of all analysed fungal spores. Negative and significant correlations were with daily mean relative humidity. The variance explained percentage by regression analyses varied between 30.6 and 39.6 % for Alternaria and Cladosporium airborne spores. Statistical methods used in this study are complementary and confirmed stable dependence of Alternaria and Cladosporium spore concentrations on meteorological factors. The climate change parameters either increased temperatures, changed precipitation regimes or a combination of both affected allergenic fungal spore concentrations in western Romania. This study demonstrates the need for investigations throughout the year, from month to month, regarding the correct interpretation of airborne spore relationships with meteorological parameters.     

Inflammation in Lafora Disease: Evolution with Disease Progression in Laforin and Malin Knock-out Mouse Models

Lafora progressive myoclonus epilepsy (Lafora disease, LD) is a fatal rare autosomal recessive neurodegenerative disorder characterized by the accumulation of insoluble ubiquitinated polyglucosan inclusions in the cytoplasm of neurons, which is most commonly associated with mutations in two genes: EPM2A, encoding the glucan phosphatase laforin, and EPM2B, encoding the E3-ubiquitin ligase malin. The present study analyzes possible inflammatory responses in the mouse lines Epm2a ^?/? (laforin knock-out) and Epm2b ^?/? (malin knock-out) with disease progression. Increased numbers of reactive astrocytes (expressing the GFAP marker) and microglia (expressing the Iba1 marker) together with increased expression of genes encoding cytokines and mediators of the inflammatory response occur in both mouse lines although with marked genotype differences. C3ar1 and CxCl10 messenger RNAs (mRNAs) are significantly increased in Epm2a ^?/? mice aged 12 months when compared with age-matched controls, whereas C3ar1, C4b, Ccl4, CxCl10, Il1b, Il6, Tnfα, and Il10ra mRNAs are significantly upregulated in Epm2b ^?/? at the same age. This is accompanied by increased protein levels of IL1-β, IL6, TNFα, and Cox2 particularly in Epm2b ^?/? mice. The severity of inflammatory changes correlates with more severe clinical symptoms previously described in Epm2b ^?/? mice. These findings show for the first time increased innate inflammatory responses in a neurodegenerative disease with polyglucosan intraneuronal deposits which increase with disease progression, in a way similar to what is seen in neurodegenerative diseases with abnormal protein aggregates. These findings also point to the possibility of using anti-inflammatory agents to mitigate the degenerative process in LD.