Immunization with V987H-stabilized Spike glycoprotein protects K18-hACE2 mice and golden Syrian hamsters upon SARS-CoV-2 infection.

Safe and effective severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines are crucial to fight against the coronavirus disease 2019 pandemic. Most vaccines are based on a mutated version of the Spike glycoprotein [K986P/V987P (S-2P)] with improved stability, yield and immunogenicity. However, S-2P is still produced at low levels. Here, we describe the V987H mutation that increases by two-fold the production of the recombinant Spike and the exposure of the receptor binding domain (RBD). S-V987H immunogenicity is similar to S-2P in mice and golden Syrian hamsters (GSH), and superior to a monomeric RBD. S-V987H immunization confer full protection against severe disease in K18-hACE2 mice and GSH upon SARS-CoV-2 challenge (D614G or B.1.351 variants). Furthermore, S-V987H immunized K18-hACE2 mice show a faster tissue viral clearance than RBD- or S-2P-vaccinated animals challenged with D614G, B.1.351 or Omicron BQ1.1 variants. Thus, S-V987H protein might be considered for future SARS-CoV-2 vaccines development.

Plitidepsin as an Immunomodulator against Respiratory Viral Infections.

Plitidepsin is a host-targeted compound known for inducing a strong anti-SARS-CoV-2 activity, as well as for having the capacity of reducing lung inflammation. Because IL-6 is one of the main cytokines involved in acute respiratory distress syndrome, the effect of plitidepsin in IL-6 secretion in different in vitro and in vivo experimental models was studied. A strong plitidepsin-mediated reduction of IL-6 was found in human monocyte-derived macrophages exposed to nonproductive SARS-CoV-2. In resiquimod (a ligand of TLR7/8)-stimulated THP1 human monocytes, plitidepsin-mediated reductions of IL-6 mRNA and IL-6 levels were also noticed. Additionally, although resiquimod-induced binding to DNA of NF-κB family members was unaffected by plitidepsin, a decrease in the regulated transcription by NF-κB (a key transcription factor involved in the inflammatory cascade) was observed. Furthermore, the phosphorylation of p65 that is required for full transcriptional NF-κB activity was significantly reduced by plitidepsin. Moreover, decreases of IL-6 levels and other proinflammatory cytokines were also seen in either SARS-CoV-2 or H1N1 influenza virus-infected mice, which were treated at low enough plitidepsin doses to not induce antiviral effects. In summary, plitidepsin is a promising therapeutic agent for the treatment of viral infections, not only because of its host-targeted antiviral effect, but also for its immunomodulatory effect, both of which were evidenced in vitro and in vivo by the decrease of proinflammatory cytokines.

Immunisation efficacy of a stabilised SARS-CoV-2 spike glycoprotein in two geriatric animal models.

Age is associated with reduced efficacy of vaccines and linked to higher risk of severe COVID-19. Here we determined the impact of ageing on the efficacy of a SARS-CoV-2 vaccine based on a stabilised Spike glycoprotein (S-29) that had previously shown high efficacy in young animals. Thirteen to 18-month-old golden Syrian hamsters (GSH) and 22-23-month-old K18-hCAE2 mice were immunised twice with S-29 protein in AddaVaxTM adjuvant. GSH were intranasally inoculated with SARS-CoV-2 either two weeks or four months after the booster dose, while all K18-hACE2 mice were intranasally inoculated two weeks after the second immunisation. Body weight and clinical signs were recorded daily post-inoculation. Lesions and viral load were investigated in different target tissues. Immunisation induced seroconversion and production of neutralising antibodies; however, animals were only partially protected from weight loss. We observed a significant reduction in the amount of viral RNA and a faster viral protein clearance in the tissues of immunized animals. Infectious particles showed a faster decay in vaccinated animals while tissue lesion development was not altered. In GSH, the shortest interval between immunisation and inoculation reduced RNA levels in the lungs, while the longest interval was equally effective in reducing RNA in nasal turbinates; viral nucleoprotein amount decreased in both tissues. In mice, immunisation was able to improve the survival of infected animals. Despite the high protection shown in young animals, S-29 efficacy was reduced in the geriatric population. Our research highlights the importance of testing vaccine efficacy in older animals as part of preclinical vaccine evaluation.

Novel Spike-stabilized trimers with improved production protect K18-hACE2 mice and golden Syrian hamsters from the highly pathogenic SARS-CoV-2 Beta variant.

Most COVID-19 vaccines are based on the SARS-CoV-2 Spike glycoprotein (S) or their subunits. However, S shows some structural instability that limits its immunogenicity and production, hampering the development of recombinant S-based vaccines. The introduction of the K986P and V987P (S-2P) mutations increases the production and immunogenicity of the recombinant S trimer, suggesting that these two parameters are related. Nevertheless, S-2P still shows some molecular instability and it is produced with low yield. Here we described a novel set of mutations identified by molecular modeling and located in the S2 region of the S-2P that increase its production up to five-fold. Besides their immunogenicity, the efficacy of two representative S-2P-based mutants, S-29 and S-21, protecting from a heterologous SARS-CoV-2 Beta variant challenge was assayed in K18-hACE2 mice (an animal model of severe SARS-CoV-2 disease) and golden Syrian hamsters (GSH) (a moderate disease model). S-21 induced higher level of WH1 and Delta variants neutralizing antibodies than S-2P in K18-hACE2 mice three days after challenge. Viral load in nasal turbinate and oropharyngeal samples were reduced in S-21 and S-29 vaccinated mice. Despite that, only the S-29 protein protected 100% of K18-hACE2 mice from severe disease. When GSH were analyzed, all immunized animals were protected from disease development irrespectively of the immunogen they received. Therefore, the higher yield of S-29, as well as its improved immunogenicity and efficacy protecting from the highly pathogenic SARS-CoV-2 Beta variant, pinpoint the S-29 mutant as an alternative to the S-2P protein for future SARS-CoV-2 vaccine development.

Contamination of As, Cd, Cr, Hg and Pb in soils in Arica commune (Chile).

The objectives of this study are (a) to determine the background concentration of As, Cd, Cr, Hg and Pb in Arica commune; (b) to determine the degree of soil contamination in Arica city using environmental indices and (c) to evaluate the human health risk of these potentially toxic elements. In the rural area of Arica commune, 169 samples were taken and 283 samples were taken in the urban area of Arica city. Total concentrations of Cd, Pb and Cr were determined by EPA 3052 and EPA 6010 C. Mercury was determined by EPA 7473. Arsenic was determined by EPA 7061A. The available concentrations of As and Cr were determined by dilute hydrochloric acid and EPA 6010C. Environmental indices were applied for pollution and US EPA model was used to evaluate human health risk. Background concentrations were As 18.2, Cd 1.12, Cr 73.2, Hg 0.02 and Pb 11.8 mg kg-1, respectively. Environmental indices show that soil samples are located between slightly contaminated to extremely contaminated. Human health risk analysis shows that children have higher levels of risk than adults. The analysis with available concentrations of As and Cr shows no carcinogenic risk for adults and children, but 81% and 98% of the samples were between 10-6 and 10-4, that means intermediate risk.

ß-Cyclodextrins as affordable antivirals to treat coronavirus infection.

The SARS-CoV-2 pandemic made evident that there are only a few drugs against coronavirus. Here we aimed to identify a cost-effective antiviral with broad spectrum activity and high safety profile. Starting from a list of 116 drug candidates, we used molecular modelling tools to rank the 44 most promising inhibitors. Next, we tested their efficacy as antivirals against α and ß coronaviruses, such as the HCoV-229E and SARS-CoV-2 variants. Four drugs, OSW-1, U18666A, hydroxypropyl-ß-cyclodextrin (HßCD) and phytol, showed in vitro antiviral activity against HCoV-229E and SARS-CoV-2. The mechanism of action of these compounds was studied by transmission electron microscopy and by fusion assays measuring SARS-CoV-2 pseudoviral entry into target cells. Entry was inhibited by HßCD and U18666A, yet only HßCD inhibited SARS-CoV-2 replication in the pulmonary Calu-3 cells. Compared to the other cyclodextrins, ß-cyclodextrins were the most potent inhibitors, which interfered with viral fusion via cholesterol depletion. ß-cyclodextrins also prevented infection in a human nasal epithelium model ex vivo and had a prophylactic effect in the nasal epithelium of hamsters in vivo. All accumulated data point to ß-cyclodextrins as promising broad-spectrum antivirals against different SARS-CoV-2 variants and distant alphacoronaviruses. Given the wide use of ß-cyclodextrins for drug encapsulation and their high safety profile in humans, our results support their clinical testing as prophylactic antivirals.

Preclinical evaluation of a COVID-19 vaccine candidate based on a recombinant RBD fusion heterodimer of SARS-CoV-2.

Current COVID-19 vaccines have been associated with a decline in infection rates, prevention of severe disease, and a decrease in mortality rates. However, SARS-CoV-2 variants are continuously evolving, and development of new accessible COVID-19 vaccines is essential to mitigate the pandemic. Here, we present data on preclinical studies in mice of a receptor-binding domain (RBD)-based recombinant protein vaccine (PHH-1V) consisting of an RBD fusion heterodimer comprising the B.1.351 and B.1.1.7 SARS-CoV-2 variants formulated in SQBA adjuvant, an oil-in-water emulsion. A prime-boost immunisation with PHH-1V in BALB/c and K18-hACE2 mice induced a CD4+ and CD8+ T cell response and RBD-binding antibodies with neutralizing activity against several variants, and also showed a good tolerability profile. Significantly, RBD fusion heterodimer vaccination conferred 100% efficacy, preventing mortality in SARS-CoV-2 infected K18-hACE2 mice, but also reducing Beta, Delta and Omicron infection in lower respiratory airways. These findings demonstrate the feasibility of this recombinant vaccine strategy.

Agreement and differential use of laboratory methods for the detection and quantification of SARS-CoV-2 in experimentally infected animals.

Rodents are widely used for the development of COVID-19-like animal models, the virological outcome being determined through several laboratory methods reported in the literature. Our objective was to assess the agreement between methods performed on different sample types from 342 rodents experimentally infected with SARS-CoV-2 (289 golden Syrian hamsters and 53 K18-hACE2 mice). Our results showed moderate agreement between methods detecting active viral replication, and that increasing viral loads determined by either RT-qPCR or infectious viral titration corresponded to increasing immunohistochemical scores. The percentage of agreement between methods decreased over experimental time points, and we observed poor agreement between RT-qPCR results and viral titration from oropharyngeal swabs. In conclusion, RT-qPCR and viral titration on tissue homogenates are the most reliable techniques to determine the presence and replication of SARS-CoV-2 in the early and peak phases of infection, and immunohistochemistry is valuable to evaluate viral distribution patterns in the infected tissues.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and humoral responses against different variants of concern in domestic pet animals and stray cats from North-Eastern Spain.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus disease 2019 (COVID-19) pandemic in humans, is able to infect several domestic, captive and wildlife animal species. Since reverse zoonotic transmission to pets has been demonstrated, it is crucial to determine their role in the epidemiology of the disease to prevent further spillover events and major spread of SARS-CoV-2. In the present study, we determined the presence of virus and the seroprevalence to SARS-CoV-2, as well as the levels of neutralizing antibodies (nAbs) against several variants of concern (VOCs) in pets (cats, dogs and ferrets) and stray cats from North-Eastern of Spain. We confirmed that cats and dogs can be infected by different VOCs of SARS-CoV-2 and, together with ferrets, are able to develop nAbs against the ancestral (B.1), Alpha (B.1.1.7), Beta (B.1.315), Delta (B.1.617.2) and Omicron (BA.1) variants, with lower titres against the latest in dogs and cats, but not in ferrets. Although the prevalence of active SARS-CoV-2 infection measured as direct viral RNA detection was low (0.3%), presence of nAbs in pets living in COVID-19-positive households was relatively high (close to 25% in cats, 10% in dogs and 40% in ferrets). It is essential to continue monitoring SARS-CoV-2 infections in these animals due to their frequent contact with human populations, and we cannot discard the probability of a higher animal susceptibility to new potential SARS-CoV-2 VOCs.

Susceptibility of Domestic Goat (Capra aegagrus hircus) to Experimental Infection with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) B.1.351/Beta Variant.

A wide range of animal species are susceptible to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Natural and/or experimental infections have been reported in pet, zoo, farmed and wild animals. Interestingly, some SARS-CoV-2 variants, such as B.1.1.7/Alpha, B.1.351/Beta, and B.1.1.529/Omicron, were demonstrated to infect some animal species not susceptible to classical viral variants. The present study aimed to elucidate if goats (Capra aegagrus hircus) are susceptible to the B.1.351/Beta variant. First, an in silico approach was used to predict the affinity between the receptor-binding domain of the spike protein of SARS-CoV-2 B.1.351/Beta variant and angiotensin-converting enzyme 2 from goats. Moreover, we performed an experimental inoculation with this variant in domestic goat and showed evidence of infection. SARS-CoV-2 was detected in nasal swabs and tissues by RT-qPCR and/or immunohistochemistry, and seroneutralisation was confirmed via ELISA and live virus neutralisation assays. However, the viral amount and tissue distribution suggest a low susceptibility of goats to the B.1.351/Beta variant. Therefore, although monitoring livestock is advisable, it is unlikely that goats play a role as SARS-CoV-2 reservoir species, and they are not useful surrogates to study SARS-CoV-2 infection in farmed animals.

Distribution of potentially harmful elements in attic dust from the City of Coronel (Chile).

Attic dusts provide an indirect measure of airborne pollutants deposited in the urban environment. The objectives of this study are: (1) to determine the concentrations of As, Ba, Cd, Co, Cr, Cu, Mn, Ni, Pb, Sr, V and Zn in attic dust in the City of Coronel, (2) to evaluate the source apportionment of PHE and (3) to assess the risk of health effects from exposure in adults and children. In the City of Coronel, attic dust samples were collected in 19 houses. The concentrations of As, Ba, Cd, Co, Cr, Cu, Mn, Ni, Pb, Sr, V and Zn were measured in ICP-OES after Aqua Regia digestion of < 75 µm dust sample. The median (and the range) concentration (mg kg-1) of potentially harmful elements was: As 16 (7-72), Ba 154 (53-251), Cd 0.8 (0.25-14.5), Co 12, (8-22), Cr 38 (22-482), Cu 107 (44-1641), Mn 698 (364-1245), Ni 51 (24-1734), Pb 66 (18-393), Sr 131 (52-252), V 129 (57-376) and Zn 815 (107-9761). The exploratory data analysis shows that Ni, Cu, Cr, Zn, Pb and As distribution is dominated by anthropogenic sources and characterized by high extreme values. Principal component analysis shows four factors. One factor is geogenic, while the other three factors are associated with transport emissions and the industrial park. The resulting median of cumulative noncarcinogenic risk (HIs) value for attic dust was 3.49 for children. This is significant, as any value greater than one indicates an elevated risk.

Potentially toxic elements concentrations in schoolyard soils in the city of Coronel, Chile.

Urban areas are constantly growing. By 2050, the urban world population, it is predicted to reach 6 billion. Being component of cities environment, urban soils have elevated levels of potentially toxic elements from anthropogenic action. The aims of this study are (1) to establish background levels of potentially toxic element in soils in the city of Coronel and (2) to assess the pollution and identify its origin. Samples (129 in total) were collected in Coronel, from 43 sites in schoolyards. Three samples were taken at each site: 0-10 cm, 10-20 cm and 150 cm depth. Principal component analysis (PCA), cluster analysis (CA) and depth ratios were applied to distinguish the origin of the contamination. The geoaccumulation index, contamination factor and the integrated pollution index were used to estimate the pollution. The median concentration of the chemical elements in 0-10 cm depth was Ba 38 mg kg-1; Co 15 mg kg-1; Cr 18 mg kg-1; Cu 22 mg kg-1; Mn 536 mg kg-1; Ni 35.5 mg kg-1; Pb 6 mg kg-1; V 94 mg kg-1; Zn 65 mg kg-1. Principal component analysis and CA suggested that Co, Ni and Mn were mainly derived from geogenic origin, while Ba, Cr, Cu, Pb, V and Zn from anthropic origin. Contamination factor indicated that some soil samples were classified as considerable contaminated to very highly contaminated by Ba, Pb, Zn and V.

First Detection of SARS-CoV-2 Delta (B.1.617.2) Variant of Concern in a Dog with Clinical Signs in Spain.

Several cases of naturally infected dogs with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been reported despite the apparently low susceptibility of this species. Here, we document the first reported case of infection caused by the Delta (B.1.617.2) variant of concern (VOC) in a dog in Spain that lived with several household members suffering from Coronavirus Infectious Disease 2019 (COVID-19). The animal displayed mild digestive and respiratory clinical signs and had a low viral load in the oropharyngeal swab collected at the first sampling. Whole-genome sequencing indicated infection with the Delta variant, coinciding with the predominant variant during the fifth pandemic wave in Spain. The dog seroconverted, as detected 21 days after the first sampling, and developed neutralizing antibodies that cross-neutralized different SARS-CoV-2 variants. This study further emphasizes the importance of studying the susceptibility of animal species to different VOCs and their potential role as reservoirs in the context of COVID-19.

Monitoring Natural SARS-CoV-2 Infection in Lions (Panthera leo) at the Barcelona Zoo: Viral Dynamics and Host Responses.

To date, no evidence supports the fact that animals play a role in the epidemiology of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus infectious disease 2019 (COVID-19). However, several animal species are naturally susceptible to SARS-CoV-2 infection. Besides pets (cats, dogs, Syrian hamsters, and ferrets) and farm animals (minks), different zoo animal species have tested positive for SARS-CoV-2 (large felids and non-human primates). After the summer of 2020, a second wave of SARS-CoV-2 infection occurred in Barcelona (Spain), reaching a peak of positive cases in November. During that period, four lions (Panthera leo) at the Barcelona Zoo and three caretakers developed respiratory signs and tested positive for the SARS-CoV-2 antigen. Lion infection was monitored for several weeks and nasal, fecal, saliva, and blood samples were taken at different time-points. SARS-CoV-2 RNA was detected in nasal samples from all studied lions and the viral RNA was detected up to two weeks after the initial viral positive test in three out of four animals. The SARS-CoV-2 genome was also detected in the feces of animals at different times. Virus isolation was successful only from respiratory samples of two lions at an early time-point. The four animals developed neutralizing antibodies after the infection that were detectable four months after the initial diagnosis. The partial SARS-CoV-2 genome sequence from one animal caretaker was identical to the sequences obtained from lions. Chronology of the events, the viral dynamics, and the genomic data support human-to-lion transmission as the origin of infection.

Protection against reinfection with D614- or G614-SARS-CoV-2 isolates in golden Syrian hamster.

Reinfections with SARS-CoV-2 have already been documented in humans, although its real incidence is currently unknown. Besides having a great impact on public health, this phenomenon raises the question of immunity generated by a single infection is sufficient to provide sterilizing/protective immunity to a subsequent SARS-CoV-2 re-exposure. The Golden Syrian hamster is a manageable animal model to explore immunological mechanisms able to counteract COVID-19, as it recapitulates pathological aspects of mild to moderately affected patients. Here, we report that SARS-CoV-2-inoculated hamsters resolve infection in the upper and lower respiratory tracts within seven days upon inoculation with the Cat01 (G614) SARS-CoV-2 isolate. Three weeks after the primary challenge, and despite high titres of neutralizing antibodies, half of the animals were susceptible to reinfection by both identical (Cat01, G614) and variant (WA/1, D614) SARS-CoV-2 isolates. However, upon re-inoculation, only nasal tissues were transiently infected with much lower viral replication than those observed after the first inoculation. These data indicate that a primary SARS-CoV-2 infection is not sufficient to elicit a sterilizing immunity in hamster models but protects against lung disease.

Accumulation of potentially toxic elements by plants of North Caucasian Alyssum species and their molecular phylogenetic analysis.

Accumulations of potentially toxic metals were investigated in soils and five North Caucasian Alyssum species from metalliferous areas and non-metalliferous areas in Karachay-Cherkessia, Kabardino-Balkaria, Dagestan and the Krasnodar region. Analyses of field samples showed that chemical features of the soils significantly affected the concentrations of Ni, Co, Zn, but had less effect on Cu and Pb concentrations in the shoots of Alyssum. Variations in the degree of accumulating ability were found in the studied species, including hyperaccumulation of Ni in Alyssum murale (up to 12,100 mg kg-1), and significant accumulation of Zn in A. gehamense (up to 1700 mg kg-1). A comparative molecular genetic analysis of two A. murale populations, both Ni-hyperaccumulating population from Karachay-Cherkessia and non-hyperaccumulating population from Dagestan, indicated considerable genetic difference between them. This result supports the hypothesis that the selection of metal hyperaccumulator species with enhanced phytoremediation efficiency should be considered at the population level.

Element Accumulation Patterns of Native Plant Species under the Natural Geochemical Stress.

A biogeochemical study of more than 20,000 soil and plant samples from the North Caucasus, Dzungarian Alatau, Kazakh Uplands, and Karatau Mountains revealed features of the chemical element uptake by the local flora. Adaptation of ore prospecting techniques alongside environmental approaches allowed the detection of geochemical changes in ecosystems, and the lessons learned can be embraced for soil phytoremediation. The data on the influence of phytogeochemical stress on the accumulation of more than 20 chemical elements by plants are considered in geochemical provinces, secondary fields of deposits, halos surrounding ore and nonmetallic deposits, zones of regional faults and schist formation, and over lithological contact lines of chemically contrasting rocks overlain by 5-20 m thick soils and unconsolidated cover. We have corroborated the postulate that the element accumulation patterns of native plants under the natural geochemical stress depend not only on the element content in soils and the characteristics of a particular species but also on the values of ionic radii and valences; with an increase in the energy coefficients of a chemical element, its plant accumulation decreases sharply. The contribution of internal factors to element uptake from solutions gives the way to soil phytoremediation over vast contaminated areas. The use of hyperaccumulating species for mining site soil treatment depends on several external factors that can strengthen or weaken the stressful situation, viz., the amount of bedrock exposure and thickness of unconsolidated rocks over ores, the chemical composition of ores and primary halos in ore-containing strata, the landscape and geochemical features of sites, and chemical element migration patterns in the supergene zone.

Relationship of the mobile forms of calcium and strontium in soils with their accumulation in meadow plants in the area of Kashin-Beck endemia.

This study was aimed at assessment of strontium and calcium mobility in soils and their accumulation with plants in the areas endemic for Kashin-Beck disease in Eastern Transbaikalia. The strontium and calcium mobility levels were determined using the method of sequential chemical extraction for 7 samples of meadow soils collected from the endemic region and 7 soil samples taken from conditionally control sites. To measure the Ca and Sr levels in the soil and plant samples, XRF analysis and AAS were used. The increased strontium level in the meadow soils of the endemic areas is accompanied by the element's higher mobility. The highest strontium yield was observed in the course of soil extraction using 1 M ammonium acetate, while the soils taken from the control sites gave lower amounts of the trace element. Furthermore, there is a positive correlation between the amount of the strontium extracted and its content in plants (r = + 0.86 - 0.98). At the sequential chemical extraction of calcium from the soils using the above method, the calcium yield was maximal in the ammonium acetate fraction (background sites) and in ammonium acetate and 6 M HCl fractions (endemic areas). The correlation between the amount of the calcium extracted in 1 M ammonium acetate and the macroelement levels found in plants was + 0.968. In addition, a peculiarly high accumulation of strontium in various willow species as compared to other meadow plants was revealed for the first time ever. Thus, the work introduces new data into the trace element biogeochemistry and environmental monitoring.

The influence of the industrial area on the pollution outside its borders: a case study from Quintero and Puchuncavi districts, Chile.

The specific objectives of this study were: (1) to establish background content levels of potentially harmful elements in topsoil of Quintero and Puchuncavi districts; (2) to assess the degree of topsoil pollution and identify the local sources of pollution; and (3) to assess the health risk of metal exposure to population. Data from a Chilean Environmental Ministry report were analyzed. Multivariate analysis suggests that the potentially harmful elements are split into two main groups, which can be explained by controlling geogenic (Ni, V and Zn) and anthropogenic (As, Cd, Cu and Pb) factors. The spatial distribution of As, Cd, Cu and Pb displays a spatial pattern around the Puchuncavi-Ventanas industrial park. The background values determined with MAD (median absolute deviation) method include As 31.6 mg kg-1, Cd 0.7 mg kg-1, Cu 296 mg kg-1, Ni 16.9 mg kg-1, Pb 48.6 mg kg-1, V 199 mg kg-1 and Zn 192 mg kg-1. The pollution indices reported considerable contamination to very high contamination in some soil samples. Hazard quotient values were higher for children than for adults, due to the ingestion and dermal absorption, while the cancer risk index values were in the acceptable range.

Prospective application of melanized fungi for the biofiltration of indoor air in closed bioregenerative systems.

Cultures of melanized fungi representative of the black yeast orders Capnodiales (Cladosporium cladosporioides and Neohortaea acidophila) and Chaetothyriales (Cladophialophora psammophila) were confined with indoor air from the laboratory during 48 h. Volatile organic compounds (VOCs) from the headspace were analyzed by thermal desorption gas chromatography time-of-fly mass spectrometry (TD-GC-ToFMS, detection threshold 0.1 µg m-3) and compared against an abiotic control. A mixture of 71 VOCs were identified and quantified in the indoor air (total concentration 1.4 mg m-3). Most of these compounds were removed in the presence of fungal biomass, but 40 newly formed putative volatile metabolites were detected, though at comparatively low total concentrations (<50 µg m-3). The VOCs emission profile of C. cladosporioides, a ubiquitous and well-known species often associated to the sick building syndrome, was consistent with previous literature reports. The specialized C. psammophila and N. acidophila, isolated respectively from gasoline polluted soil and from lignite, displayed rather specific VOCs emission profiles. Mass balances on the fungal uptake and generation of VOCs resulted in overall VOCs removal efficiencies higher than 96% with all tested fungi. Applied aspects and biosafety issues concerning the suitability of black yeasts for the biofiltration of indoor air have been discussed.