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Bee pollen is consumed as a dietary supplement. Its quality parameters are regulated by the Argentine Food Code (AFCode). The present study characterized 10 commercial dehydrated samples from five Argentine provinces to provide information on hygienic quality and health safety. We assessed their microbiological quality, including potential mycotoxins. We also determined their botanical origin and moisture. Results showed that seven out of ten samples presented lower counts of culturable heterotrophic mesophilic bacteria than those allowed by the AFCode. In contrast, all samples showed higher filamentous fungi and yeast counts than the level approved by the AFCode. No fumonisin or deoxynivalenol was detected; we observed only aflatoxin B2 in one sample and ochratoxin A in two. The results of the botanical origin of samples showed that all samples had a predominant pollen type, except one. The microbiological quality of all samples agreed with that required by the AFCode, except filamentous fungi and yeasts. In addition, we found variations among samples from the same province. Therefore, if pollen is intended for human consumption, appropriate hygiene standards must be applied to all bee pollen production operations. Highlights: • The microbiological quality of bee pollen samples, regardless of the province or botanical origin, agreed with the requirements of the AFCode, except for filamentous fungus and yeast. • All samples showed filamentous fungus and yeast counts higher than the permitted level in the AFCode. • Mycotoxin levels found in bee pollen were far from the limits allowed in humans (10 µg.kg-1) by regulations for other foods. • All samples analyzed were pollen mixes verified by identifying numerous botanical species. • The characterization of Argentinian commercial bee pollen showed that it was suitable for human consumption.

The antimicrobial activity of four concentrations of hydroalcoholic extracts from honeybee propolis and Scaptotrigona jujuyensis geopropolis was screened in vitro against five tomato pathogenic bacteria. The agar-well diffusion method was used and the tested bacteria were Clavibacter michiganensis subsp. michiganensis, Xanthomonas gardneri, Xanthomonas vesicatoria, Pseudomonas corrugata, and Pseudomonas mediterranea. The main chemical characteristics of propolis and geopropolis, including the polyphenol profile through HPLC-DAD, were also determined. Geopropolis raw sample presented higher values of moisture (7.78%), waxes (50.79%) and ashes (3.69%) than propolis (4.59%, 31.16% and 2.42% respectively). The total polyphenol content and the dry extract were higher in propolis hydroalcoholic extract (3.83 mg eq galic acid mL-1 and 7.87%, respectively) than in the extract of geopropolis (0.16 mg eq galic acid mL-1 and 0.15%, respectively). Chromatographic analysis confirmed the presence of caffeic acid, quercetin, 1,5,7-trihydroxy-flavanone, apigenin, pinobanksin, chrysin, pinocembrin, and galangin in both extracts. The antimicrobial assay showed significant differences between the hydroalcoholic extract activities, as well as between the sensitivity of the tested bacteria. Propolis hydroalcoholic extract dilutions had an inhibitory effect over four of the five tested bacteria, while geopropolis hydroalcoholic extract dilutions were only effective against C. michiganensis subsp. michiganensis, and to a lesser extent. The sequence of bacteria sensitivity to propolis treatments was: C. michiganensis subsp. michiganensis > X. gardneri > X. vesicatoria > P. corrugata. Pseudomonas mediterranea was not sensitive to any of the hydroalcoholic extracts. The antimicrobial activity of both extracts was dose-dependent where the most concentrated treatments were the most effective (15.0 mg mL?1 of geopropolis and 78.7 mg mL?1 of propolis dry extract, respectively). The polyphenol content and the HPLC-DAD profile of the hydroalcoholic extracts disclosed differences in chemical composition that helped to explain the outcomes of the in vitro assay. These results are a contribution to the characterization of bee bioactive products, specifically to propolis and geopropolis. This study indicates the likelihood of using propolis as a non-conventional strategy to control tomato bacterial diseases.

La transformación de los compuestos de fósforo orgánico (Po) a fósforo inorgánico (Pi) soluble, es denominada mineralización y es llevada a cabo por un grupo de enzimas conocidas como fosfatasas. En este trabajo, se estudió la actividad fosfatasa de cinco lotes de la región pampeana norte del área sojera argentina, mediante: la evaluación de la actividad fosfatasa del suelo, y el recuento de las comunidades bacterianas y fúngicas con esa actividad, y de esta manera se obtuvo información sobre el potencial de los mismos para movilizar el Po. Se determinó el número de bacterias aeróbicas heterotróficas cultivables (BAHC) así como el de hongos cultivables (HC), además del número de productores de fosfatasas ácidas y alcalinas. El número de bacterias con actividad fosfatasa fue en promedio 6,85 10(5) UFC g-1 de suelo para la fosfatasa ácida; mientras que para la fosfatasa alcalina fue en promedio 5,80 10(5) UFC g-1 de suelo. En cambio, el valor medio de hongos con actividad fosfatasa ácida fue 1,78 10³ UFC g-1 de suelo y para la enzima alcalina 1,77 10³ UFC g-1 de suelo. No se encontraron diferencias estadísticamente significativas entre el número de bacterias y el de hongos con fosfatasa ácida o alcalina entre los distintos suelos. Por otro lado, el nivel de actividad de la fosfatasa alcalina osciló entre 5,72 y 15,5 mg p-nitrofenol kg-1 suelo h-1, mientras la ácida varió entre 27,4 y 105 mg p-nitrofenol kg-1 suelo h-1. Se encontraron diferencias significativas en la actividad enzimática entre los cinco suelos, siendo mayor la actividad ácida que la alcalina. Los resultados de este trabajo demostraron que los suelos estudiados presentan una actividad mineralizadora de fuentes de Po que coincide con otros trabajos de suelos cultivados y que el recuento de los organismos productores de fosfatasas complementa la información obtenida a partir de la determinación de la actividad fosfatasa del suelo. Mediante la utilización de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po.Transformation of organic phosphorus (Po) into soluble inorganic phosphorus (Pi) is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB) and cultivated fungi (CF) was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5) CFU g-1 soil, while alkaline activity was 5.80 10(5) CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5) mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

En el presente trabajo se estudia la habilidad fisiológica para solubilizar fosfato inorgánico de diferentes grupos bacterianos así como de cepas de Bradyrhizobium sp. aislados de suelos sojeros. Se recolectaron muestras de suelo y se determinó el número de la microflora total así como el total de bacterias, hongos y de solubilizadores. La capacidad de solubilizar fosfato tricálcico de los grupos de bacterias predominantes en el suelo, de 250 cepas de Bradyrhizobium sp. y de 10 cepas de colección se probó en placa de Petri conteniendo el medio NBRIP con 5 g L -1 de fosfato tricálcico. Se midió el tamaño de los halos de solubilización y se aislaron aquellas colonias que mostraban halos mayores a los 4 mm. El fósforo solubilizado se estimó cuantitativamente en medio líquido con y sin agregado de solución tampón y se comparó con una cepa comercial. El valor medio de bacterias totales fue 5,1 106 (0,06% solubilizadoras) mientras que el valor medio de hongos totales alcanzó a 3.3 104 (9.70% solubilizadores). No se encontraron diferencias significativas en el número de bacterias solubilizadoras de fosfato en los distintos suelos. Se obtuvieron 14 aislamientos solubilizadores: 10 de la microflora y 4 de Bradyrhizobium sp. y sus halos oscilaron entre 4 y 15 mm. Todas las cepas de colección excepto MSDJ G 49 solubilizaron fosfato en placa de Petri. Las cantidades de fosfato solubilizadas por las bacterias de la microflora variaron entre 3% y 24,1% en un medio líquido sin solución tamponada mientras que en un medio tamponado variaron entre 0,07% y 4,82%. En un medio líquido sin solución reguladora, los aislamientos de Bradyrhizobium sp. solubilizaron en porcentajes que variaron entre 7,1% y 8,5% mientras que en un medio tamponado oscilaron entre 0,1% y 0,16%. No se observaron diferencias significativas en las cantidades de fosfato solubilizadas por los aislamientos de Bradyrhizobium en un medio tamponado con respecto a la mayoría de los aislamientos de la microflora. No se destacó la capacidad de solubilización de fosfato de la cepa comercial con respecto a lo observado en los aislamientos de este trabajo. Nuestros resultados permiten concluir que existen aislamientos de bacterias del suelo con diferente capacidad solubilizadora de fósforo inorgánico y que las cepas de Bradyrhizobium sp. muestran capacidades de solubilización comparables a las de otros géneros bacterianos.The present research focuses on the study of the ability to solubilize inorganic phosphate of different bacterial groups as well as of Bradyrhizobium sp. strains isolated from soybean soils. Soil samples were collected and the microflora as well as the total number of bacteria, fungi, and solubilizers was assessed. The phosphate solubilization ability of the predominant bacterial groups, of 250 Bradyrhizobium sp. strains, and of 10 collection strains was tested in Petri dishes containing NBRIP supplemented with 5 g L-1 of tricalcium phosphate. The size of solubilizing halos was measured and those colonies showing halos larger than 4 mm were isolated. Solubilized phosphate was quantitavely estimated in broth with and without buffer in these strains and it was compared with a commercial strain. The total bacteria mean values reached 5.1 106 (0.06% solubilizers) while the total fungi reached 3.3 104 (9.70% solubilizers). No significant differences in the number of solubilizing bacteria were observed in the different soils. Fourteen solubilizing strains were obtained: 10 from microflora and 4 Bradyrhizobium sp. strains whose halos ranged from 4 to 15 mm. All collection strains except MSDJ G49 solubilized phosphate in the plate assay. The amounts of solubilized phosphate by the strains of microflora varied from 3% to 24.1% in a liquid medium without buffer while with buffer varied from 0.07% to 4.82%. In a liquid medium without buffer, the strains of Bradyrhizobium sp. solubilized in percentages ranging from 7.1% to 8.5% while with buffer, the percentages ranged from 0.1% to 0.16%. No significant differences were observed in the amounts of solubilized phosphate in a liquid medium with buffer between strains of Bradyrhizobium sp. and the majority of the microflora strains. The phosphate solubilization ability of the commercial strain studied was not as relevant as that of the other strains. Our results lead to conclude that there are bacterial strains with different abilities to solubilize inorganic phosphate and that the strains of Bradyrhizobium sp. show relevant solubilization ability with respect to other bacterial genera.

Dekkera bruxellensis is one of the most important contaminant yeasts of alcoholic fermentation. The use of propolis, which can selectively target contaminating yeasts without affecting the starter one, Saccharomyces cerevisiae, could be a useful nonconventional strategy for controlling the growth of contaminant yeasts. The objective of this research was to evaluate four samples of propolis produced by Apis mellifera honeybees from different regions of Argentina as antimicrobial agents against the growth of D. bruxellensis and S. cerevisiae. Hydroalcoholic extracts of propolis were prepared with ethanol:water (70:30 v/v), and the specific absorbance and final concentration of the samples were evaluated. A qualitative in vitro assay in solid medium was performed with different propolis concentrations, and the evaluation of yeast growth was based on a qualitative scale. A quantitative in vitro assay in liquid medium was also performed to assess the yeast cell number, using two different propolis concentrations. The cell number of D. bruxellensis decreased 1.52 and 1.85 log cycles with the two propolis extracts utilised at a concentration of 4.5 mg mL-1 while the cell number of S. cerevisiae decreased 0.48 and 0.76 log cycles with the same samples of propolis. The results from both assays demonstrated the selectivity of propolis use on the yeast species, leading to a higher inhibition of D. bruxellensis growth. This indicates a good potential for using propolis at the concentration of 4.5 mg mL-1, as a nonconventional strategy to control the growth of D. bruxellensis without significantly affecting S. cerevisiae, the yeast starter of ethanol fermentation.