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Fusarioid fungi, particularly Neocosmospora solani and Fusarium oxysporum, are emerging as significant human pathogens, causing infections ranging from localized mycoses to life-threatening systemic diseases. Accurate identification and preservation of these fungi in clinical laboratories remain challenging because of their diverse morphologies and specific growth requirements. This study evaluated a novel milk-honey and malt agar (MHM) against conventional media for cultivating and preserving 60 clinical fusarioid isolates, including Neocosmospora spp. (n = 47), Bisifusarium spp. (n = 5), and Fusarium spp. (n = 8). Compared with Sabouraud dextrose 2 % agar (SDA) and malt extract agar (ME2), MHM significantly increased conidia production (p < 0.0001, mean = 3.4 × 103, standard deviation (SD) = ±1.3 × 103), with results similar to those of carnation leaf agar (CLA). MHM facilitated superior preservation of fusarioid viability for up to one year at room temperature on slant cultures and over two years on swabs in Amies gel with charcoal, outperforming current methods such as Castellani (water) or cryopreservation. Morphological characterization of fusarioid fungi grown on MHM revealed distinct growth patterns and conidial structures for Neocosmospora, Bisifusarium, and Fusarium species, aiding in identifying these genera. The superior performance of MHM in stimulating conidiation, maintaining viability, and preserving morphology underscore its potential as a reference medium for medically relevant fusarioid fungi, with broad implications for clinical mycology laboratories and resource-limited settings.

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Introduction: Sporotrichosis is a subcutaneous mycosis caused by fungi of the genus Sporothrix sp. Phenotypic and genotypic differences have been associated with their geographic distribution, virulence, or clinical manifestation of sporotrichosis. In the past decade, the interest in identifying species of the Sporothrix sp. has been increasing, due to its epidemiological importance and, in consequence, is important to know how to preserve them for future studies, in culture collection. Aims: The purposes of this study were to analyze the global distribution of environmental isolates and/or causal agents of sporotrichosis identified by polyphasic taxonomy, with mandatory use of molecular identification, and to evaluate the percentages and distribution of isolates stored in culture collections. Methods: A systematic review of articles on animal and human sporotrichosis and/or environmental isolation of the fungus, from 2007 to 2023, was done. Results: Our results demonstrated that, S. globosa, S. schenckii, and S. brasiliensis were the most identified species. With respect to the deposit and maintenance of species, we observed that only 17% of the strains of Sporothrix sp. isolated in the world are preserved in a culture collection. Conclusions: This systematic review confirmed a difficulty in obtaining the frequency of Sporothrix species stored in culture collection and insufficient data on the molecular identification mainly of animal sporotrichosis and isolation of Sporothrix sp. in environmental samples.

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SUMMARY: Osteotechnics is one of the different anatomical preservation techniques and can be defined as the technique designed to prepare, clean, obtain and preserve bone structures that can be used in the teaching, museographic or research field. The osteotechnical technique procedure consists of the following phases: debulk and disjoint, maceration, cooking, cleaning, degreasing, bleaching, and labeling to obtain bone material. Seven phases will be explained in detail, as well as the materials, instruments, quantities of the substances used, and the time required to obtain human bone material. We consider that this article can serve as a guide, given that all the experimentation was carried out with human biological material. This methodological proposal could be consolidated and established based on the experience acquired during the creation of the contemporary skeletal collection of the department of innovation in human biological material (DIMBIH). Therefore, the purpose of our proposal is to provide tools that facilitate the work of those who carry out this work and fundamentally to avoid irreversible or irreparable damage to the osteological material, since it is of great value and difficult to acquire for disciplines as anatomy, veterinary, physical and forensic anthropology, medicine, dentistry and biology.

La osteotecnia es una de las técnicas diferentes de conservación anatómica y puede definirse como la técnica destinada a preparar, limpiar, obtener y conservar estructuras óseas que pueden ser utilizadas en el ámbito docente, museográfico o de investigación. El procedimiento de la técnica osteotécnica consta de las siguientes fases: descarnado y desarticulado, maceración, cocción, limpieza, desengrase, blanqueo y marcaje para la obtención de material óseo. Se explicarán en detalle siete fases, así como los materiales, instrumentos, cantidades de las sustancias utilizadas y el tiempo necesario para obtener material óseo humano. Consideramos que este artículo puede servir de guía, dado que toda la experimentación se realizó con material biológico humano. Esta propuesta metodológica pudo consolidarse y establecerse a partir de la experiencia adquirida durante la creación de la colección esquelética contemporánea del Departamento de Innovación en Material Biológico Humano (DIMBIH). Por lo tanto, el propósito de nuestra propuesta es brindar herramientas que faciliten el trabajo de quienes realizan este trabajo y fundamentalmente evitar daños irreversibles o irreparables en el material osteológico, ya que es de gran valor y difícil adquisición para las disciplinas como la anatomía, veterinaria, antropología física y forense, medicina, odontología y biología.

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The efficiency of alternative preservation techniques for Xanthomonas arboricola pv pruni was studied. The preservation methods in sunflower seeds, glass beads and sterile soil were suitable for maintaining viability and productive capacity of xanthan pruni.

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Different regions have different environmental conditions, which may be unfavorable for the preservation of the quality of stored soybean seeds over time. Thus, it is necessary to adopt specific technologies to control the storage environment conditions. Big raffia bags are widely used for the storage of soybean seeds, however these consist of a porous, permeable material that allows the exchange of gases between the packaging and the storage environment. In an effort to find a solution to this problem, in this study we evaluated low cost big bag coating alternatives, in order to minimize the effects of temperature and intergranular humidity on stored seeds. Thus, the aim of this work was to evaluate the quality of soybean cultivars subjected to different temperature and storage duration conditions and stored in raffia bags with or without internal coating. We used a completely randomized, three-factor (10 × 6 × 5) experimental design. We assessed 10 soybean cultivars, six storage environments, and five evaluation periods. Our results showed that seeds of the M-SOY 8866, M7110 IPRO, CD 2737 RR, and BMX DESAFIO 8473 RSF soybean cultivars preserved their physiological quality better in different storage environments. The storage duration had a cumulative effect on the negative factors that favor the deterioration of the quality of the stored seeds. The storage temperature was the main factor that affected the physiological quality of the stored seeds. The use of coated packaging was beneficial in preserving the physiological quality of stored soybean seeds; however, its effect was greater at ambient temperature than in a cold environment. The best storage environment for the preservation of the quality of the seeds was characterized by 10°C temperature conditions and the use of coated packaging, while the worst storage environment was characterized by ambient temperature conditions without the use of coated packaging. Thus, it was concluded that the use of coatings in raffia big bags can be an alternative for maintaining the quality of seeds of different soybean cultivars during storage in seed processing units.

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Background: Freeze-drying is known as one of the best methods to preserve bacterial strains. Protectant is the key factor affecting the survival rate of freeze-dried strains. In addition, salinity, bacterial suspension concentration, drying time, and other factors can also affect the survival rate of strains to varying degrees. At present, there are relatively few studies on freeze-drying preservation of marine bacteria. In the present study, we performed the freeze-drying protectant screening and optimized the preservation conditions for Pseudoalteromonas nigrifaciens, which is widely distributed in marine environment. The protective effects of the screened protectants were verified by 18 other marine bacterial strains. Results: The results indicated that the combination of 5.0% (w/v) lactose, 5.0% (w/v) mannitol, 5.0% (w/v) trehalose, 10.0% (w/v) skim milk powder, 0.5% (w/v) ascorbic acid and 0.5% (w/v) gelatin was the best choice for the preservation of P. nigrifaciens. The suggested salinity and concentration of initial cell suspension were 10 g/L NaCl and 1.0 × 109 CFU/mL, respectively. Furthermore, stationary-phase cells were the best choice for the freeze-drying process. The highest survival rate of P. nigrifaciens reached 52.8% when using 5­10% (w/v) skim milk as rehydration medium. Moreover, the other 18 marine strains belonging to Pseudoalteromonas, Vibrio, Photobacterium, Planomicrobium, Edwardsiella, Enterococcus, Bacillus, and Saccharomyces were freezedried under the abovementioned conditions. Their survival rates were 2.3­95.1%. Conclusion: Collectively, our results supported that the protectant mixture and parameters were beneficial for lyophilization of marine bacteria

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Reference fungal cultures (RFCs) are essential for the internal quality control of laboratories. The production of these cultures requires standardized procedures (IRAM 14950:2016 and ISO 17034:2016 standards) carried out by a recognized and accredited laboratory. The aim of this work was to produce RFC in paper disks of autochthonous strains, characterized by two, homogeneous and stable reference methods traceable at species level. RFC were produced using 14 regional species (7 yeasts and 7 filamentous fungi) from the fungal culture collection (DMic). Paper disks were impregnated with a culture suspension, dried and packed. Homogeneity, viability, identity and purity were verified. Short-and long-term stability at different temperatures and storage times were studied. Characterization of each strain allowed to confirm its identity and to ensure its traceability at international level. Produced batches were homogeneous and stable at -20 ±5 °C for 30 months. This method of production was adequate to produce homogeneous and stable RFC with phenotypic and genotypic characteristics correctly defined and internationally traceable. Standardized procedures were developed for the production of certified RFC that could be transferred to other microorganisms. Providing RFC that represent regional strains allows laboratories to produce more reliable results with a favorable impact on medical diagnosis, the environment or the food industry.

Los cultivos microbianos de referencia (CR) son imprescindibles para el control de calidad interno de los laboratorios. Asegurar su producción requiere de procedimientos estandarizados (IRAM 14950:2016 e ISO 17034:2016) realizados en un laboratorio reconocido y acreditado. El objetivo de este estudio fue producir cultivos fúngicos de referencia en discos de papel, a partir de un panel de cultivos autóctonos caracterizados por dos métodos de referencia, trazables a nivel taxonómico de especie, homogéneos y estables. Se produjeron CR de 14 especies circulantes en Argentina (7 de levaduras y 7 de hongos miceliales), depositadas en la colección de hongos de interés médico (DMic). Los discos de papel fueron embebidos con una suspensión del cultivo por producir, secados y envasados. Se verificó la homogeneidad, viabilidad, identidad y pureza de cada lote. Se evaluó la estabilidad a corto y largo plazo a distintas temperaturas y tiempos de almacenamiento. La caracterización de cada CR nos permitió confirmar su identidad y asegurar su trazabilidad a nivel internacional. Los lotes producidos fueron homogéneos y estables durante 30 meses conservados a -20 ±5 °C. Este método resultó adecuado para producir CR homogéneos y estables, con características fenotípicas y genotípicas correctamente definidas y trazables a nivel internacional. Los procedimientos estandarizados desarrollados en este trabajo pueden ser transferidos para producir CR certificados de otros microorganismos. La provisión de CR que represente cepas regionales permite a los laboratorios producir resultados más confiables con un impacto favorable en el diagnóstico médico, los estudios ambientales y la industria alimenticia.

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Frente a necessidade de preservação e a manutenção de materiais biológicos, dentre eles fungos com potencial para controle biológico, para o desenvolvimento biotecnológico e científico, que vêm ganhando destaque no cenário mundial. Sendo necessário a adequação de métodos de preservação que além de garantia a sobrevivência destes microrganismos permitam a conservação de suas características morfológicas, fisiológicas e genéticas, no entanto, não existe um método ideal ou universal para a conservação de materiais biológicos. Diante desta necessidade o presente trabalho teve como objetivo de avaliar a eficácia e viabilidade de três métodos de preservação de isolados do fungo Phomadimorpha (repicagens periódicas, Castellani e fragmentos de papel-filtro), em dois períodos de avaliação, seis e doze meses após o armazenamento. Estudou-se a eficácia e viabilidade, através do crescimento micelial do fungo em meio de cultivo contendo batata-dextrose-ágar. Houve variabilidade entre os métodos de preservação do isolado do fungo P. dimorpha para o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial, nos dois períodos de avaliação, após seis e doze meses de armazenamento. O método de preservação em fragmento de papel filtro mostrou-se como o mais eficaz na preservação do isolado do fungo P. dimorpha nos dois períodos de avaliação, após seis e doze meses de armazenamento, sendo ideal para obter o maior o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial.

Facing the need for preservation and maintenance of biological materials, among them fungi with potential for biological control, for biotechnological and scientific development, which are gaining prominence in the world scenario. It is necessary to adapt preservation methods that besides guaranteeing the survival of these microorganisms allow the conservation of their morphological, physiological and genetic characteristics, however, there is no ideal or universal method for the conservation of biological materials. In view of this need, the present work had the objective of evaluating the efficacy and feasibility of three methods for the preservation of Phoma dimorpha (periodic transfer, Castellani and filter paper fragments) isolates in two evaluation periods, six and twelve months after the storage. Efficacy and viability were studied by mycelial growth of the fungus in a culture medium containing potato-dextrose-agar. There was variability between the preservation methods of the P. dimorpha fungus isolate for mycelial growth, method efficacy and mycelial growth rate index, in the two evaluation periods, after six and twelve months of storage. The filter paper fragment preservation method was the most effective in reserving the P. dimorpha fungus isolate in the two evaluation periods, after six and twelve months of storage, being ideal to obtain the highest mycelial growth, efficacy of the method and mycelial growth rate index.

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Frente a necessidade de preservação e a manutenção de materiais biológicos, dentre eles fungos com potencial para controle biológico, para o desenvolvimento biotecnológico e científico, que vêm ganhando destaque no cenário mundial. Sendo necessário a adequação de métodos de preservação que além de garantia a sobrevivência destes microrganismos permitam a conservação de suas características morfológicas, fisiológicas e genéticas, no entanto, não existe um método ideal ou universal para a conservação de materiais biológicos. Diante desta necessidade o presente trabalho teve como objetivo de avaliar a eficácia e viabilidade de três métodos de preservação de isolados do fungo Phomadimorpha (repicagens periódicas, Castellani e fragmentos de papel-filtro), em dois períodos de avaliação, seis e doze meses após o armazenamento. Estudou-se a eficácia e viabilidade, através do crescimento micelial do fungo em meio de cultivo contendo batata-dextrose-ágar. Houve variabilidade entre os métodos de preservação do isolado do fungo P. dimorpha para o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial, nos dois períodos de avaliação, após seis e doze meses de armazenamento. O método de preservação em fragmento de papel filtro mostrou-se como o mais eficaz na preservação do isolado do fungo P. dimorpha nos dois períodos de avaliação, após seis e doze meses de armazenamento, sendo ideal para obter o maior o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial.(AU)

Facing the need for preservation and maintenance of biological materials, among them fungi with potential for biological control, for biotechnological and scientific development, which are gaining prominence in the world scenario. It is necessary to adapt preservation methods that besides guaranteeing the survival of these microorganisms allow the conservation of their morphological, physiological and genetic characteristics, however, there is no ideal or universal method for the conservation of biological materials. In view of this need, the present work had the objective of evaluating the efficacy and feasibility of three methods for the preservation of Phoma dimorpha (periodic transfer, Castellani and filter paper fragments) isolates in two evaluation periods, six and twelve months after the storage. Efficacy and viability were studied by mycelial growth of the fungus in a culture medium containing potato-dextrose-agar. There was variability between the preservation methods of the P. dimorpha fungus isolate for mycelial growth, method efficacy and mycelial growth rate index, in the two evaluation periods, after six and twelve months of storage. The filter paper fragment preservation method was the most effective in reserving the P. dimorpha fungus isolate in the two evaluation periods, after six and twelve months of storage, being ideal to obtain the highest mycelial growth, efficacy of the method and mycelial growth rate index.(AU)

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The effects of full-fat goat's milk and/or inulin and/or oligofructose, as carrier agents, were investigated to improve the survival rates of Bifidobacterium BB-12, and the physical properties of the microcapsules under storage conditions. On the day of their manufacture, the microcapsules were evaluated for morphology, particle size, and distribution of fat and bifidobacteria. The viability of the bifidobacteria, moisture and fat content, water activity, solubility, bulk and tapped density, flowability, cohesiveness and color properties were evaluated for 120 days at 4 °C and 25 °C. The full-fat goat's milk powder with or without inulin as encapsulating agents showed the highest survival rates of Bifidobacterium BB-12 after spray drying and storage. Considering the bifidobacteria survival, both of these spray-dried powders showed the most desirable physical properties, i.e., lowest water activity and solubility, respectively. Both properties are highlighted for better stability of spray-dried powders, with microcapsules, during storage time. These results are credited to full-fat goat's milk (200 g L-1) and the presence of inulin (100 g L-1), besides the fat content showing a high correlation with the solubility values. The lowest volume occupied by the spray-dried powders was noted when oligofructose was used as the carrier agent. The samples that showed the presence of cracks influenced negatively on the bifidobacteria viability. These cracks were responsible by the greater water escape, resulting in powders with more desirable lower water activity. In relation to the color parameters, lower stability was noted when oligofructose was used, while the best stability was also noted for the powders with full-fat goat's milk and/or inulin. During storage time, the best performance was achieved by the microencapsulation process that used only full-fat goat's milk and/or inulin and storage at 4 °C.

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RATIONALE: Distinct techniques employed to preserve different types of tissues may affect stable isotope analyses conducted on samples, and this is critical when field work takes place in remote areas. To investigate this, the stable isotope ratios (δ13 C and δ15 N values) obtained using two methods commonly used to preserve humpback whale (and other cetaceans) skin samples were compared. METHODS: A total of 54 pairs of skin samples of humpback whales from the southern Baja California Peninsula, Mexico, were preserved in ethanol (90%) and by freezing, between 2007 and 2009. The δ13 C and δ15 N values were determined using a PDZ Europa ANCA-GSL elemental analyzer interfaced to a PDZ Europe 20-20 isotope ratio mass spectrometer. Parametric and nonparametric tests were used to compare the isotopic results. RESULTS: A significant (t = 4.93; p = 0.000003) variation of −0.92‰ was found between the mean δ13 C values in ethanol (from −19.38‰ to −16.07‰; mean = −17.86‰) and freezing (from −20.67‰ to −16.44‰; mean = −18.78‰) techniques. No significant (U = 1314, p = 0.38) differences were observed in the δ15 N values. The δ13 C values were compared between preservation methods for each of the three years under analysis. Significant differences were observed in 2007 (t = 3.45; p = 0.0012) and 2008 (t = 3.13; p = 0.0030), but not for 2009 (t = 1.66; p = 0.12). CONCLUSIONS: Based on the results of this study, the use of ethanol to preserve humpback whale skin samples collected for stable isotope analysis is not recommended, particularly regarding the analysis of δ13 C values. This study serves as a point of reference for future research on humpback whales or other whales involving skin samples preserved by freezing or in ethanol.

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The oral administration of non-toxigenic strains of Clostridioides difficile (NTCD) is currently showing promising results for the prevention of Clostridioides difficile infection (CDI) in humans and animals, and is being considered as a possible commercial product to be used in the near future. The aim of this work was to evaluate five culture media for the growth and sporulation of one NTCD (Z31) and evaluate the viability of a lyophilized spore solution of NTCD Z31 stored at 4 °C or at 25 °C for 2 years. Reinforced clostridial medium (RCM) and brain heart infusion broth (BHI) provided the highest production of NTCD Z31 spores. In the first 6 months of the storage of the lyophilized solution, a reduction in spore count of approximately 0.3 Log10 CFU/mL was observed; however, no further significant reduction in spore count was observed up to 24 months. No difference in spore concentration was found between the two storage temperatures from 6 to 24 months of storage. The present work showed BHI and RCM to be the best choices for the growth and sporulation of NTCD Z31 and suggested that the spores of NTCD Z31 are stable for up to 2 years under both temperature conditions.

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Reference fungal cultures (RFCs) are essential for the internal quality control of laboratories. The production of these cultures requires standardized procedures (IRAM 14950:2016 and ISO 17034:2016 standards) carried out by a recognized and accredited laboratory. The aim of this work was to produce RFC in paper disks of autochthonous strains, characterized by two, homogeneous and stable reference methods traceable at species level. RFC were produced using 14 regional species (7 yeasts and 7 filamentous fungi) from the fungal culture collection (DMic). Paper disks were impregnated with a culture suspension, dried and packed. Homogeneity, viability, identity and purity were verified. Short- and long-term stability at different temperatures and storage times were studied. Characterization of each strain allowed to confirm its identity and to ensure its traceability at international level. Produced batches were homogeneous and stable at -20±5°C for 30 months. This method of production was adequate to produce homogeneous and stable RFC with phenotypic and genotypic characteristics correctly defined and internationally traceable. Standardized procedures were developed for the production of certified RFC that could be transferred to other microorganisms. Providing RFC that represent regional strains allows laboratories to produce more reliable results with a favorable impact on medical diagnosis, the environment or the food industry.

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RESUMEN: El auge experimentado en los últimos años en la aplicación de las técnicas anatómicas para la conservación de muestras anatómicas está directamente relacionado con la necesidad de preservación de los escasos especímenes con que cuentan las instituciones universitarias en relación a aumentar el tiempo de utilización del mismo. En este sentido, la plastinación es la técnica anatómica que más se destaca y que permite preservar por tiempo indeterminado, sin toxicidad, las preparaciones anatómicas. Presentamos el protocolo modificado de plastinación a temperatura ambiente con silicona, desarrollado en el Laboratorio de Plastinación y Técnicas Anatómicas de la Universidad de La Frontera, con el objetivo de aplicarla a la conservación de una placenta humana, la cual posteriormente fue pigmentada para otorgarle un aspecto más cercano a lo real.

SUMMARY: The surge experienced in recent years in the application of anatomical techniques for the conservation of anatomical samples is directly related to the need to preserve the few specimens that university institutions have in relation to increase the time of use of the same. In this sense, the plastination is the anatomical technique that stands out and that allows to preserve indefinitely, without toxicity, the anatomical preparations. We present the modified plastination protocol at room temperature with silicone, developed in the Laboratory of Plastination and Anatomical Techniques of the University of La Frontera, with the aim of applying it to the conservation of a human placenta, which was subsequently pigmented to give it an appearance closer to the real.

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SummaryIn this study we analyzed whether the in vivo storage of oocytes (time after ovulation until fertilization) affects the survival and the ploidy status of the yellowtail tetra Astyanax altiparanae. Fish were induced to spawn and, after ovulation, a small aliquot was stripped and immediately fertilized (positive control group). Subsequently, aliquots (~150 oocytes) were stripped and fertilized at various time points of 60, 120, 180 or 240 min. Developmental stages, abnormalities, survival and the ploidy status of the hatched larvae were examined. As expected, in the control group, 100% of the larvae were diploid. Conversely, triploid individuals were observed just at the 60 min treatment time point (0.6%). In vivo storage of oocytes also influenced the survival rates (P < 0.05); the 180 and 240 min samples, respectively, presented lower survival rates at gastrula (50.10±6.26% and 40.92±5.32%), and somite (17.80±5.14% and 4.41±2.76%) stages and lower hatching rates (12.01±4.04% and 4.41±2.76%). A higher percentage (99.27±0.40%) of normal larvae and only a few abnormal larvae (0.73±0.40%) were observed in the control group (P = 0.0000). This observation did not differ from that observed at the 60 min treatment point (P = 0.9976). A significant increase in the percentage of abnormalities was observed in the other treatments, and, after 240 min, the highest percentage of abnormal larvae was seen (P=0.0024; 83.33±16.67%). In conclusion, we showed that oocyte ageing had a significant effect on survival and may affect the ploidy status in A. atiparanae.

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Preservation methods for entomopathogenic fungi (EPF) require effective protocols to ensure uniform processes and to avoid alterations during storage. The aim of this study was to preserve Beauveria bassiana, Metarhizium acridum, M. anisopliae, M. rileyi, Isaria javanica, Hirsutella thompsonii, H. citriformis and Lecanicillium lecanii in mineral oil (MO), sterile water (SW), silica gel (SG), lyophilisation (L), ultracold-freezing at -70 °C, and cryopreservation at -196 °C. The viability and purity of the fungi were then verified: phenotypic characteristics were evaluated qualitatively at 6, 12 and 24 m. Genetic stability was tested by amplified fragment length polymorphisms (AFLP) analysis at 24 m. Of the eight species of EPF, three remained viable in SW, five in MO and L, six at -70 °C, seven in SG, and eight at -196 °C. No significant changes were observed in AFLP patterns at 24 m of storage. The most effective preservation methods for EPF were SG, L, -70 and -196 °C. Beauveria bassiana, M. acridum, M. anisopliae, M. rileyi and I. javanica remained stable with all methods, while the remaining species were less compatible. The optimisation of preservation methods for EPF facilitates the development of reliable protocols to ensure their inherent characteristics in culture collections.

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Formaldehyde has been prominent in preserving biological tissues since the nineteenth century. Despite being admittedly harmful to health and to the environment, it is still widely used. The Morphology Department of the University of Brasília - Brazil, applied the rethink, reduce, reuse, recycle and responsibility methodology to their activities in an effort to protect the health of laboratory workers and users, save resources and reduce damage to the environment. Here we evaluate the results obtained a decade after the implementation of this proposal (2005-2015). Formaldehyde was replaced by alcohol and glycerol solutions in corpse conservation. Over five thousand dollars in public funds that would have been destined to buying preserving substances were saved annually, and over a hundred thousand liters of water that would have been contaminated and thrown into the sewage system were spared. The environment used to implement the study was improved and anatomical parts kept for study had their lifespan extended. It is noteworthy that such simple adjustments could cause pronounced changes in laboratory activities. We would avoid contaminating billions of liters of water and it would be possible to save millions if similar practices were implemented in all educational institutions having similar routines.

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Strains of Agaricus brasiliensis require special procedures for conservation. Thus, the objective of this research was to establish conservation and maintenance procedures A. brasiliensis strain PSWC838 from the University of Pennsylvania (ABWC838) and an A. brasiliensis strain from the Fujian Agriculture and Forestry University (ABC). The medium in which mycelia grew the quickest for both strains was selected using a multifactorial design with 2 strains, 4 culture media, and incubation for 5 different times; the growth rate (mm/day) was the response variable. Analysis of variance showed that the potato dextrose agar (PDA) medium and potato extract did not display a significantly different growth rate, and PDA was selected for safety reasons. We also evaluated the viability of the strains grown on PDA and 0.2% activated carbon after 3 months of storage in sterile distilled water. A factorial design was applied with 2 factors, the strain and incubation for 10 different times. The Tukey post hoc test indicated that ABC showed quicker and more homogeneous growth than ABWC838. Finally, the results showed that pieces of mycelium of ABC and ABWC838 strains inoculated on the PDA medium with 0.2% activated carbon and preserved in sterile distilled water at 18 ± 1°C showed 100% viability after 3 months of storage. Moreover, the results of semiquantitative biochemical tests confirmed that the production of laccases and amylases was preserved in these strains after storage in sterile water, enhancing their ability to degrade substrates containing lignin and starchy waste.

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Anaerobic ammonium oxidation (anammox) bacteria have peculiar characteristics that make them difficult to cultivate. The conservation of these microorganisms in culture collections or laboratories requires successful preservation and reactivation techniques. Furthermore, studies have shown that successful reactivation may be preservative dependent. Considering this, the present study aimed to evaluate the preservation and reactivation of anammox consortia enriched from swine manure treatment lagoons, by using different preservative agents at different temperatures: KNO3 (at 4 °C), glycerol (-20 °C, -80 °C), and skimmed cow milk (-20 °C, -80 °C, -200 °C). After 4 months, the biomass was thawed (except for KNO3), and the reestablishment of anammox activity was evaluated by stoichiometric coefficients. Microbial community transformation during the reactivation process was also studied by 16S rDNA sequence analysis. The results showed that the anammox biomass preserved with glycerol or skimmed cow milk at -80 °C recovered activity, while the biomass preserved with other methodologies did not reestablish activity during the studied time (90 days). The bacterial community from the biomass with anammox activity was characterized and showed the presence of Candidatus Brocadia anammoxidans, Candidatus Jettenia asiatica, and Candidatus Anammoxoglobus propionicus. Preservation with skimmed cow milk at -80 °C favored the selection of Candidatus Anammoxoglobus propionicus, while preservation with glycerol at -80 °C was successful for Candidatus Jettenia asiatica. The present study was effective on anammox sludge preservation and reactivation using low-cost processes for anammox cultures preservation, which is important for biomass transport and deammonification reactor start up.

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Phlebotomine (Diptera, Psychodidae, Phlebotominae) taxonomy has been studied extensively, primarily due to the role of these flies as vectors of various parasites, including species of Leishmania, Bartonella and arboviruses that cause diseases in humans and other vertebrates. We present some topics discussed at a round-table on phlebotomine taxonomy held at the Ninth International Symposium on Phlebotomine Sandflies (ISOPS IX) in Reims, France, in June 2016. To date, approximately one thousand phlebotomine species have been described worldwide, although in varying languages and mostly without standardization of characters and terminology. In the interest of standardization, we list the characters that should minimally be considered in the description of new phlebotomine taxa as well as annotated illustrations of several characters. For these characters, multiple illustrations are provided to show some of the variations. The preferred terms for all pertinent characters are listed as well as their synonyms in English, Portuguese, and French. Finally, we offer an updated list of abbreviations to be used for generic and subgeneric names.

Asunto(s)