Exploring fungal diversity in Antarctic wildlife: isolation and molecular identification of culturable fungi from penguins and pinnipeds.

AIMS: To survey the diversity of fungal species that may be cultured from Antarctic penguins and pinnipeds, and to test the in vitro susceptibility to triazole drugs of any medically important Aspergillus spp. isolates. METHODS: During an expedition to Argentinean Antarctic research stations at Potter Peninsula (South Shetland Islands) and Primavera Cape (Antarctic Peninsula) in February 2019, samples (n = 212) were collected from fur seals (Arctocephalus gazella), elephant seals (Mirounga leonine), leopard seals (Hydrurga leptonyx), Weddell seals (Leptonychotes weddellii) and crabeater seals (Lobodon carcinophaga) and gentoo penguins (Pygoscelis papua). Oral, nasal and rectal swabs and skin/hair brushings were collected from pinnipeds, and skin/feather brushings, cloacal swabs and moulted feathers from penguins. Samples were cultured on Sabouraud dextrose agar and/or potato dextrose agar plates and fungal isolates identified by morphological criteria followed by PCR amplification and DNA sequencing. Antifungal susceptibility of Aspergillus spp. isolates to triazoles was tested. RESULTS: Fungi from 21 genera were isolated from 121/212 (57.1%) samples obtained from pinnipeds and penguins. Among pinnipeds from Potter Peninsula (fur seals and elephant seals), the most frequent fungal species were Debaryomyces hansenii and Rhodotorula mucilaginosa, isolated from the oral, nasal and/or rectal mucosa, and Antarctomyces psychrotrophicus isolated from the skin/hair of all sampled individuals. Among pinnipeds from Primavera Cape (leopard seals, Weddell seals and crabeater seals), the most frequent fungal species were Naganishia adeliensis and Cryptococcus neoformans var. uniguttulatus, isolated from the nasal/oral mucosa of 4/33 (15.2%) and 5/33 (12.1%) animals, respectively. The most frequently isolated fungal species from gentoo penguins (Potter Peninsula), were Pseudogymnoascus pannorum and A. pyschrotrophicus, which both were isolated from skin/feathers of 7/15 (46.7%) birds, and Thelebolus microsporus, isolated from the cloacal mucosa and skin/feathers of 5/15 (33.3%) and 2/15 (13.3%) birds, respectively. Fungi that are potentially pathogenic to both humans and animals (Aspergillus fumigatus, Asp. flavus, Asp. versicolor, Candida parapsilosis and Microsporum canis) were isolated from 4/38 (10.5%), 1/38 (2.6%), 2/38 (5.3%), 4/38 (10.5%) and 2/38 (5.3%) sampled pinnipeds, respectively. Only non-azole-resistant isolates of Asp. fumigatus and Asp. flavus were identified. CONCLUSIONS: The fungal biodiversity in Antarctic pinnipeds and gentoo penguins was explored using standard mycological culture followed by PCR and DNA sequencing. The frequency of fungal carriage varied among animal species, sample type and location. This study constitutes an epidemiologic approach to monitoring of these marine animals for emerging fungal pathogens.

Low levels of PAHs and organotin compounds in surface sediment samples from a broad marine area of 25 de Mayo (King George) Island, South Shetland Islands.

The Northern region of the Antarctic Peninsula constitutes the area with the highest human presence in West Antarctica. The human presence, with all the activities associated such as logistic, scientific and tourism operations, represents a potential risk of chemical pollution with both, organic and inorganic contaminants. Under these conditions knowledge about the presence and levels of the main persistent organic pollutants (POPs) is essential to evaluate the environmental status of this ecologically relevant and sensitive area. In this work, which complements our previous study regarding trace elements, we performed the first regional-scale monitoring of 24 PAHs (16 of them included in EPA list of primary pollutant), and organotin compounds (OTCs:TBT, DBT and MBT) in surface sediment from 68 sites comprising six different areas in Maxwell Bay, southeast coast of 25 de Mayo (King George) Island. POPs were quantified in surface sediment samples (20-30 m depth) obtained during two summer Antarctic expeditions by gas chromatography-mass spectrometry (GC-MS). The two most anthropized areas (South Fildes and Potter Cove) showed moderated evidence of pollution for PAHs and OTC. In some sampling sites the concentration of total PAHs was higher than 100 ng/g dw, while TBT was detected in only five samples, two of them located in Potter Cove (ranged between 14 and 18 ng/g dw), and three, located in South Fildes area (ranged between 118 and 416 ng/g dw). Although POPs contamination was evidenced in some samples close to scientific stations, a pollution pattern was not clearly identified.

Degradation of macroalgal detritus in shallow coastal Antarctic sediments.

Glaciers along the western Antarctic Peninsula are retreating at unprecedented rates, opening up sublittoral rocky substrate for colonization by marine organisms such as macroalgae. When macroalgae are physically detached due to storms or erosion, their fragments can accumulate in seabed hollows, where they can be grazed upon by herbivores or be degraded microbially or be sequestered. To understand the fate of the increasing amount of macroalgal detritus in Antarctic shallow subtidal sediments, a mesocosm experiment was conducted to track 13C- and 15N-labeled macroalgal detritus into the benthic bacterial, meiofaunal, and macrofaunal biomass and respiration of sediments from Potter Cove (King George Island). We compared the degradation pathways of two macroalgae species: one considered palatable for herbivores (the red algae Palmaria decipiens) and other considered nonpalatable for herbivores (the brown algae Desmarestia anceps). The carbon from Palmaria was recycled at a higher rate than that of Desmarestia, with herbivores such as amphipods playing a stronger role in the early degradation process of the Palmaria fragments and the microbial community taking over at a later stage. In contrast, Desmarestia was more buried in the subsurface sediments, stimulating subsurface bacterial degradation. Macrofauna probably relied indirectly on Desmarestia carbon, recycled by bacteria and microphytobenthos. The efficient cycling of the nutrients and carbon from the macroalgae supports a positive feedback loop among bacteria, microphytobenthos, and meiofaunal and macrofaunal grazers, resulting in longer term retention of macroalgal nutrients in the sediment, hence creating a food bank for the benthos.

Element concentrations of environmental concern in surface sediment samples from a broad marine area of 25 de Mayo (King George) Island, South Shetland Islands.

Western Antarctica (WA) constitutes the area with the highest human presence in the white continent and also the region where the effects of global warming are more evident worldwide. Such human presence represents a potential risk of pollution with both, organic and inorganic contaminants. Global warming also could modify dynamics and transport of the pollutants, increasing summer water runoff, ice melting and iceberg scouring. Under this fast-changing scenario, knowledge about the concentration of contaminants is essential to evaluate the environmental status of this ecologically relevant area. In this work, we performed the first regional-scale monitoring of 9 trace elements (Cr, Co, Ni, Cu, Zn, As, Pb, Cd and Hg), as well as Fe and Mn, in surface sediment from 64 sites comprising six different areas in Maxwell Bay, 25 de Mayo (King George) Island. Target elements were quantified in surface sediment samples (20-30 m depth) obtained during two summer Antarctic expeditions: 2010/11 and 2011/12 by inductively coupled plasma linked to a quadrupole mass spectrometer (ICP-MS). Based on the average values observed for the reference areas, baseline values were defined for the studied region. A regional enrichment in Cu (compared with the global mean upper crust) was observed and related to the widespread mineralization of volcanic rocks. The most anthropized area (South Fildes) mainly showed sediment class 3 (moderately polluted) for Pb, Cd and Hg with a number samples revealing some highly contaminated hot spots. Although elemental contamination in some samples close to scientific stations or sites where logistic operations were evidenced, a pollution pattern was not clearly identified. The present work represents the first regional-scale attempt to define the baseline values and the anthropic impacts in this region of the WA and also provides the first data about Hg concentration in surface sediment of the study area.

Pelagic and benthic communities of the Antarctic ecosystem of Potter Cove: Genomics and ecological implications.

Molecular technologies are more frequently applied in Antarctic ecosystem research and the growing amount of sequence-based information available in databases adds a new dimension to understanding the response of Antarctic organisms and communities to environmental change. We apply molecular techniques, including fingerprinting, and amplicon and metagenome sequencing, to understand biodiversity and phylogeography to resolve adaptive processes in an Antarctic coastal ecosystem from microbial to macrobenthic organisms and communities. Interpretation of the molecular data is not only achieved by their combination with classical methods (pigment analyses or microscopy), but furthermore by combining molecular with environmental data (e.g., sediment characteristics, biogeochemistry or oceanography) in space and over time. The studies form part of a long-term ecosystem investigation in Potter Cove on King-George Island, Antarctica, in which we follow the effects of rapid retreat of the local glacier on the cove ecosystem. We formulate and encourage new approaches to integrate molecular tools into Antarctic ecosystem research, environmental conservation actions, and polar ocean observatories.

Bioremediation of hydrocarbon-contaminated soils in cold regions: Development of a pre-optimized biostimulation biopile-scale field assay in Antarctica.

Bioremediation proved to be an effective approach to deal with soil contamination, especially in isolated, cold environments such as Antarctica. Biostimulation, involving the addition of macronutrients -mainly nitrogen and phosphorous- is considered the simplest and cheapest bioremediation process. Optimizing the levels of these nutrients is a key step prior to the application of a biostimulation strategy. In this work, N and P levels, optimized by Response Surface Methodology (RSM) at lab-scale, were applied to an Antarctic hydrocarbon contaminated soil. The process was performed on-site, using high density polyethylene geomembranes (800µm) to isolate treated soil from the surroundings and under environmental conditions at Carlini station (Antarctica) during 50days. Two 0.5ton biopiles were used as experimental units; a control biopile (CC), and a biostimulated system (BS), amended with N and P. At the end of the assay, hydrocarbon removal was significantly higher in BS system compared to CC (75.79% and 49.54% respectively), showing that the applied strategy was effective enough to perform a field-assay in Antarctica that significantly reduce soil contamination levels; and proving that RSM represents a fundamental tool for the optimization of nutrient levels to apply during bioremediation of fuel contaminated cold soils.

Fungal Planet description sheets: 154-213.

Novel species of microfungi described in the present study include the following from South Africa: Camarosporium aloes, Phaeococcomyces aloes and Phoma aloes from Aloe, C. psoraleae, Diaporthe psoraleae and D. psoraleae-pinnatae from Psoralea, Colletotrichum euphorbiae from Euphorbia, Coniothyrium prosopidis and Peyronellaea prosopidis from Prosopis, Diaporthe cassines from Cassine, D. diospyricola from Diospyros, Diaporthe maytenicola from Maytenus, Harknessia proteae from Protea, Neofusicoccum ursorum and N. cryptoaustrale from Eucalyptus, Ochrocladosporium adansoniae from Adansonia, Pilidium pseudoconcavum from Greyia radlkoferi, Stagonospora pseudopaludosa from Phragmites and Toxicocladosporium ficiniae from Ficinia. Several species were also described from Thailand, namely: Chaetopsina pini and C. pinicola from Pinus spp., Myrmecridium thailandicum from reed litter, Passalora pseudotithoniae from Tithonia, Pallidocercospora ventilago from Ventilago, Pyricularia bothriochloae from Bothriochloa and Sphaerulina rhododendricola from Rhododendron. Novelties from Spain include Cladophialophora multiseptata, Knufia tsunedae and Pleuroascus rectipilus from soil and Cyphellophora catalaunica from river sediments. Species from the USA include Bipolaris drechsleri from Microstegium, Calonectria blephiliae from Blephilia, Kellermania macrospora (epitype) and K. pseudoyuccigena from Yucca. Three new species are described from Mexico, namely Neophaeosphaeria agaves and K. agaves from Agave and Phytophthora ipomoeae from Ipomoea. Other African species include Calonectria mossambicensis from Eucalyptus (Mozambique), Harzia cameroonensis from an unknown creeper (Cameroon), Mastigosporella anisophylleae from Anisophyllea (Zambia) and Teratosphaeria terminaliae from Terminalia (Zimbabwe). Species from Europe include Auxarthron longisporum from forest soil (Portugal), Discosia pseudoartocreas from Tilia (Austria), Paraconiothyrium polonense and P. lycopodinum from Lycopodium (Poland) and Stachybotrys oleronensis from Iris (France). Two species of Chrysosporium are described from Antarctica, namely C. magnasporum and C. oceanitesii. Finally, Licea xanthospora is described from Australia, Hypochnicium huinayensis from Chile and Custingophora blanchettei from Uruguay. Novel genera of Ascomycetes include Neomycosphaerella from Pseudopentameris macrantha (South Africa), and Paramycosphaerella from Brachystegia sp. (Zimbabwe). Novel hyphomycete genera include Pseudocatenomycopsis from Rothmannia (Zambia), Neopseudocercospora from Terminalia (Zambia) and Neodeightoniella from Phragmites (South Africa), while Dimorphiopsis from Brachystegia (Zambia) represents a novel coelomycetous genus. Furthermore, Alanphillipsia is introduced as a new genus in the Botryosphaeriaceae with four species, A. aloes, A. aloeigena and A. aloetica from Aloe spp. and A. euphorbiae from Euphorbia sp. (South Africa). A new combination is also proposed for Brachysporium torulosum (Deightoniella black tip of banana) as Corynespora torulosa. Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Molybdate reduction to molybdenum blue by an Antarctic bacterium.

A molybdenum-reducing bacterium from Antarctica has been isolated. The bacterium converts sodium molybdate or Mo6⁺ to molybdenum blue (Mo-blue). Electron donors such as glucose, sucrose, fructose, and lactose supported molybdate reduction. Ammonium sulphate was the best nitrogen source for molybdate reduction. Optimal conditions for molybdate reduction were between 30 and 50 mM molybdate, between 15 and 20°C, and initial pH between 6.5 and 7.5. The Mo-blue produced had a unique absorption spectrum with a peak maximum at 865 nm and a shoulder at 710 nm. Respiratory inhibitors such as antimycin A, sodium azide, potassium cyanide, and rotenone failed to inhibit the reducing activity. The Mo-reducing enzyme was partially purified using ion exchange and gel filtration chromatography. The partially purified enzyme showed optimal pH and temperature for activity at 6.0 and 20°C, respectively. Metal ions such as cadmium, chromium, copper, silver, lead, and mercury caused more than 95% inhibition of the molybdenum-reducing activity at 0.1 mM. The isolate was tentatively identified as Pseudomonas sp. strain DRY1 based on partial 16s rDNA molecular phylogenetic assessment and the Biolog microbial identification system. The characteristics of this strain would make it very useful in bioremediation works in the polar and temperate countries.

Contact dermatitis caused by dimethylfumarate in Argentina.

For the first time in Argentina, we describe an outbreak of contact dermatitis. New pairs of shoes caused intense pruritus, pain, and eruption, followed by edema, blisters, and a severe negative impact on the epidermal barrier of the feet. We identify dimethylfumarate as the causal agent and suggest an analytical method for its fast identification.

Influence of nutrients addition and bioaugmentation on the hydrocarbon biodegradation of a chronically contaminated Antarctic soil.

Complexity involved in the transport of soils and the restrictive legislation for the area makes on-site bioremediation the strategy of choice to reduce hydrocarbons contamination in Antarctica. The effect of biostimulation (with N and P) and bioaugmentation (with two bacterial consortia and a mix of bacterial strains) was analysed by using microcosms set up on metal trays containing 2.5 kg of contaminated soil from Marambio Station. At the end of the assay (45 days), all biostimulated systems showed significant increases in total heterotrophic aerobic and hydrocarbon-degrading bacterial counts. However, no differences were detected between bioaugmented and nonbioaugmented systems, except for J13 system which seemed to exert a negative effect on the natural bacterial flora. Hydrocarbons removal efficiencies agreed with changes in bacterial counts reaching 86 and 81% in M10 (bioaugmented) and CC (biostimulated only) systems. Results confirmed the feasibility of the application of bioremediation strategies to reduce hydrocarbon contamination in Antarctic soils and showed that, when soils are chronically contaminated, biostimulation is the best option. Bioaugmentation with hydrocarbon-degrading bacteria at numbers comparable to the total heterotrophic aerobic counts showed by the natural microflora did not improve the process and showed that they would turn the procedure unnecessarily more complex.

Bacterial community dynamics during bioremediation of diesel oil-contaminated Antarctic soil.

The effect of nutrient and inocula amendment in a bioremediation field trial using a nutrient-poor Antarctic soil chronically contaminated with hydrocarbons was tested. The analysis of the effects that the treatments caused in bacterial numbers and hydrocarbon removal was combined with the elucidation of the changes occurring on the bacterial community, by 16S rDNA-based terminal restriction fragment length polymorphism (T-RFLP) typing, and the detection of some of the genes involved in the catabolism of hydrocarbons. All treatments caused a significant increase in the number of bacteria able to grow on hydrocarbons and a significant decrease in the soil hydrocarbon content, as compared to the control. However, there were no significant differences between treatments. Comparison of the soil T-RFLP profiles indicated that there were changes in the structure and composition of bacterial communities during the bioremediation trial, although the communities in treated plots were highly similar irrespective of the treatment applied, and they had a similar temporal dynamics. These results showed that nutrient addition was the main factor contributing to the outcome of the bioremediation experiment. This was supported by the lack of evidence of the establishment of inoculated consortia in soils, since their characteristic electrophoretic peaks were only detectable in soil profiles at the beginning of the experiment. Genetic potential for naphthalene degradation, evidenced by detection of nahAc gene, was observed in all soil plots including the control. In treated plots, an increase in the detection of catechol degradation genes (nahH and catA) and in a key gene of denitrification (nosZ) was observed as well. These results indicate that treatments favored the degradation of aromatic hydrocarbons and probably stimulated denitrification, at least transiently. This mesocosm study shows that recovery of chronically contaminated Antarctic soils can be successfully accelerated using biostimulation with nutrients, and that this causes a change in the indigenous bacterial communities and in the genetic potential for hydrocarbon degradation.

Extracellular proteases from the Antarctic marine pseudoalteromonas sp. P96-47 strain.

The extracellular protease-production capacity of 33 bacterial isolates taken from marine biotopes in King George Island, Antarctica, was evaluated in liquid cultures. The P96-47 isolate was selected due to its high production capacity and was identified as Pseudoalteromonas sp. The optimal growth temperature was 20 degrees C and the optimal for protease production was 15 degrees C. Proteases were purified from culture supernatants, developing a multiple-band profile in zymograms. They were classified as neutral metalloproteases and worked optimally at 45 degrees C with an Eact of 47 kJ/mol. Their stability was higher at neutral pH, retaining more than 80% of activity at pH 6-10 after 3 h incubation at 4 degrees C. After 90 min incubation at 40 and 50 degrees C, the percentages of residual activities were 78% and 44%. These results contribute to the basic knowledge of Antarctic marine proteases and also help evaluate the probable industrial applications of P96-47 proteases.

Extracellular proteases from the Antarctic marine Pseudoalteromonas sp. P96-47 strain / Proteasas extracelulares de la cepa marina antártica Pseudoalteromonas sp. P96-47

The extracellular protease-production capacity of 33 bacterial isolates taken from marine biotopes in King George Island, Antarctica, was evaluated in liquid cultures. The P96-47 isolate was selected due to its high production capacity and was identified as Pseudoalteromonas sp. The optimal growth temperature was 20 °C and the optimal for protease production was 15 °C. Proteases were purified from culture supernatants, developing a multiple-band profile in zymograms. They were classified as neutral metalloproteases and worked optimally at 45 °C with an Eact of 47 kJ/ mol. Their stability was higher at neutral pH, retaining more than 80% of activity at pH 6-10 after 3 h incubation at 4 °C. After 90 min incubation at 40 and 50 °C, the percentages of residual activities were 78% and 44%. These results contribute to the basic knowledge of Antarctic marine proteases and also help evaluate the probable industrial applications of P96-47 proteases.

La capacidad productora de proteasas extracelulares de 33 aislamientos bacterianos tomados de biotopos marinos en la Isla Rey Jorge, Antártida, fue evaluada en cultivo líquido. El aislamiento P96-47 fue seleccionado debido a su alta capacidad productora y fue identificado como Pseudoalteromonas sp. La temperatura óptima de crecimiento fue de 20 °C y la de producción de 15 °C. Las proteasas fueron purificadas a partir del sobrenadante de cultivo, y en los zimogramas desarrollaron un perfil de múltiples bandas. Estas proteasas fueron clasificadas como metaloproteasas neutras y se observó que trabajan óptimamente a 45 °C, con una Eact de 47 kJ/ mol. Su estabilidad fue superior a pH neutro y retuvieron más del 80% de su actividad a pH 6-10 después de 3 h de incubación a 4 °C. Luego de 90 min de incubación a 40 y 50 °C, las actividades residuales fueron 78% y 44%, respectivamente. Los resultados que se presentan en este trabajo contribuyen al conocimiento básico de las proteasas marinas antárticas y también a evaluar las probables aplicaciones industriales de las proteasas de P96-47.

[Changes in viability of two Antarctic marine bacteria exposed to solar radiation in the water column: influence of vertical mixing]. / Cambios en la viabilidad de dos bacterias marinas antárticas expuestas a la radiación solar en la columna de agua: influencia de la mezcla vertical.

The effect of UV radiation on two Antarctic marine bacterial strains (UVps and UVvi) was studied in the water column of Potter Cove (South Shetland, Antarctica). Quartz flasks were filled with the bacterial suspensions and exposed to solar radiation at 0 m, 1 m and 3 m depth. Assays using flasks exposed to direct solar radiation and others using flasks covered with/by interferential filters which discriminate between UVA and UVB, were performed. In other assays, a vertical mixing of 4 m/h was simulated. Both strains showed a significant decrease in viability (expressed as colony - forming units) when exposed to a surface UVB dose of 8.4 kJ m(-2). Studies with interferential filters showed a significant decrease at 0 and 1 m depth under both UV treatments. The UVps strain appeared to be more sensitive to UVB than to UVA. Damage produced by UVB was attenuated by the vertical mixing when the surface UVB dose was 4.8 kJ m(-2). This effect was not observed when surface UVB dose was 7.7 kJ m(-2). These results show that the negative effect caused by UVB radiation on the bacterioplankton would be significant only in the first meter of water column of the Antarctic coastal waters with high levels of suspended particulate material.

Cambios en la viabilidad de dos bacterias marinas antárticas expuestas a la radiación solar en la columna de agua: influencia de la mezcla vertical / Changes in viability of two Antarctic marine bacteria exposed to solar radiation in the water column: influence of vertical mixing

Se estudió el efecto de la radiación ultravioleta (RUV) sobre dos cepas bacterianas marinas antárticas (UVps y UVvi) en la columna de agua de la caleta Potter (Shetland del Sur, Antártida). Frascos de cuarzo con las cepas en estudio fueron expuestos a la radiación solar en superficie, a 1 m y a 3 m de profundidad. Se realizaron ensayos con exposición directa y con filtros interferenciales que discriminaron la radiación UVA y la UVB. En otros ensayos se simuló una mezcla vertical de 4 m/h. Ambas cepas mostraron una disminución significativa del número de unidades formadoras de colonias, tanto en superficie como a 1 m de profundidad, luego de exponerlas a dosis superficiales de UVB de 8,4 kJ m-2. El estudio con filtros interferenciales mostró una disminución significativa de la viabilidad en ambos tratamientos UV en superficie y a 1 m. La cepa UVps mostró mayor sensibilidad a la UVB que a la UVA. La mezcla vertical amortiguó el daño causado por la UVB cuando la dosis en superficie fue de 4,8 kJ m-2. Este efecto amortiguador no se observó cuando la dosis en superficie fue de 7,7 kJ m-2. Estos resultados muestran que el efecto negativo de la RUV sobre el bacterioplancton sería particularmente importante en el primer metro de profundidad de las aguas costeras antárticas con abundante material particulado en suspensión.

The effect of UV radiation on two Antarctic marine bacterial strains (UVps and UVvi) was studied in the water column of Potter Cove (South Shetland, Antarctica). Quartz flasks were filled with the bacterial suspensions and exposed to solar radiation at 0 m, 1 m and 3 m depth. Assays using flasks exposed to direct solar radiation and others using flasks covered with/by interferential filters which discriminate between UVA and UVB, were performed. In other assays, a vertical mixing of 4 m/h was simulated. Both strains showed a significant decrease in viability (expressed as colony - forming units) when exposed to a surface UVB dose of 8.4 kJ m-2. Studies with interferential filters showed a significant decrease at 0 and 1 m depth under both UV treatments. The UVps strain appeared to be more sensitive to UVB than to UVA. Damage produced by UVB was attenuated by the vertical mixing when the surface UVB dose was 4.8 kJ m-2. This effect was not observed when surface UVB dose was 7.7 kJ m-2. These results show that the negative effect caused by UVB radiation on the bacterioplankton would be significant only in the first meter of water column of the Antarctic coastal waters with high levels of suspended particulate material.

Factors influencing protease production by two Antarctic strains of Stenotrophomonas maltophilia.

The influence of culture medium buffer capacity, the supplementation of culture medium with L-ala and the requirement of calcium for exoprotease production by Antarctic psychrotrophic Stenotrophomonas maltophilia strains ANT-1-1 and ANT-7-1 were examined. When increasing concentrations of calcium chloride (0 to 0.3 g l-1) were added to culture media, maximum protease production yields increased 70-75% (ANT-1-1) and 50% (ANT-7-1), while biomass levels showed little difference. Calcium was also necessary for optimal activity of proteases. L-ala had no effect on protease production. The reduction in buffer capacity, with the consequent change in external pH, had a positive effect, enhancing protease yields. Secretion of proteases into the medium started at the beginning of the stationary phase, corresponding with a rise in pH values up to pH 8.7 and was maximal at 36 h of culture. These results indicate that the regulation of calcium concentration and buffer capacity and also pH monitoring are factors to be considered when the design of an industrial culture medium and the optimisation of protease production processes using these Antarctic strains are concerned.

Factors influencing protease production by two Antarctic strains of Stenotrophomonas maltophilia

The influence of culture medium buffer capacity, the supplementation of culture medium with L-ala and the requirement of calcium for exoprotease production by Antarctic psychrotrophic Stenotrophomonas maltophilia strains ANT-1-1 and ANT-7-1 were examined. When increasing concentrations of calcium chloride (0 to 0.3 g l-1) were added to culture media, maximum protease production yields increased 70-75 (ANT-1-1) and 50 (ANT-7-1), while biomass levels showed little difference. Calcium was also necessary for optimal activity of proteases. L-ala had no effect on protease production. The reduction in buffer capacity, with the consequent change in external pH, had a positive effect, enhancing protease yields. Secretion of proteases into the medium started at the beginning of the stationary phase, corresponding with a rise in pH values up to pH 8.7 and was maximal at 36 h of culture. These results indicate that the regulation of calcium concentration and buffer capacity and also pH monitoring are factors to be considered when the design of an industrial culture medium and the optimisation of protease production processes using these Antarctic strains are concerned.

Factors influencing protease production by two Antarctic strains of Stenotrophomonas maltophilia

The influence of culture medium buffer capacity, the supplementation of culture medium with L-ala and the requirement of calcium for exoprotease production by Antarctic psychrotrophic Stenotrophomonas maltophilia strains ANT-1-1 and ANT-7-1 were examined. When increasing concentrations of calcium chloride (0 to 0.3 g l-1) were added to culture media, maximum protease production yields increased 70-75 (ANT-1-1) and 50 (ANT-7-1), while biomass levels showed little difference. Calcium was also necessary for optimal activity of proteases. L-ala had no effect on protease production. The reduction in buffer capacity, with the consequent change in external pH, had a positive effect, enhancing protease yields. Secretion of proteases into the medium started at the beginning of the stationary phase, corresponding with a rise in pH values up to pH 8.7 and was maximal at 36 h of culture. These results indicate that the regulation of calcium concentration and buffer capacity and also pH monitoring are factors to be considered when the design of an industrial culture medium and the optimisation of protease production processes using these Antarctic strains are concerned.(AU)