Bacterial diversity using metagenomics of 16s rDNA in water kefir, an innovative source of probiotics for bee nutrition.

Water kefir is a sparkling, slightly acidic fermented beverage made from sugar, water, and water kefir grains, which are a mixture of yeast and bacteria. These grains produce a variety of fermentation compounds such as lactic acid, acetaldehyde, acetoin, ethanol and carbon dioxide. In this study, a high-throughput sequencing technique was used to characterize the bacterial composition of the original water kefir from which potential probiotics were obtained. We studied the bacterial diversity of both water kefir grains and beverages. DNA was extracted from three replicate samples of both grains and beverages using the Powerlyzer Microbial Kit. The hypervariable V1-V2 region of the bacterial 16S ribosomal RNA gene was amplified to prepare six DNA libraries. Between 1.4M and 2.4M base-pairs were sequenced for the library. In total, 28721971 raw reads were obtained from all the samples. Estimated species richness was higher in kefir beverage samples compared to grain samples. Moreover, a higher level of microbial alpha diversity was observed in the beverage samples. Particularly, the predominant bacteria in beverages were Anaerocolumna and Ralstonia, while in grains Liquorilactobacillus dominated, with lower levels of Leuconostoc and Oenococcus. Although the bacterial diversity in kefir grains was low because only three genera were the most represented, all of them are LAB bacteria with the potential to serve as probiotics in the artificial feeding of bees.

Microbiological and Chemical characterization of water kefir: An innovative source of potential probiotics for bee nutrition / Caracterización microbiológica y química del kéfir de agua: una fuente innovadora de potenciales probióticos para la nutrición de las abejas

Abstract We evaluated the microbial composition of water kefir grains and beverage overthe course of one year to determine whether the number and type of microorganisms changedover the time. Bacteria and yeast colonies with different morphologies were isolated fromwater kefir and their antimicrobial activity was evaluated against Paenibacillus larvae andAscosphaera apis. A chemical characterization of kefir was also carried out. Our results con-firmed that bacteria and yeasts were more numerous in kefir grains compared with those in thebeverage. The counts of microorganisms declined, although an important microbial community was still present in kefir after the long storage period. Eleven strains which inhibited bee pathogens were isolated from kefir. Genotypic results demonstrated that these isolates included Lentilactobacillus hilgardii, Lentilactobacillus buchneri and Saccharomyces cerevisiae. Thus, water kefir may be an innovative source of potential probiotic strains for bee nutrition in order to control honeybee diseases.

Resumen Evaluamos la composición microbiana del kéfir de agua durante un ano para determinar si la cantidad y el tipo de microorganismos cambiaban con el tiempo. Se aislaron colonias de bacterias y de levaduras con diferentes morfologías, y su actividad antimicrobiana se evaluó frente a Paenibacillus larvaey Ascosphaeraapis. También se realizó una caracterización química del kéfir. Nuestros resultados confirmaron que las bacterias y las levaduras eran más numerosas en los gránulos de kéfir en comparación con la parte líquida. Los recuentos de microorganismos disminuyeron, aunque una cantidad igualmente importante se encontró en el kéfir después de un año. Se aislaron del kéfir once cepas que inhibieron los mencionados patógenos de abejas. Los resultados genotípicos demostraron que estos aislamientos eran Lentilactobacillus hilgardii, Lentilactobacillus buchneri y Saccharomyces cerevisiae. Por lo tanto, el kéfir de agua podría ser una fuente innovadora de potenciales cepas probióticas para contribuir a la nutrición y sanidad de las abejas.

Microbiological and chemical characterization of water kefir: An innovative source of potential probiotics for bee nutrition.

We evaluated the microbial composition of water kefir grains and beverage over the course of one year to determine whether the number and type of microorganisms changed over the time. Bacteria and yeast colonies with different morphologies were isolated from water kefir and their antimicrobial activity was evaluated against Paenibacillus larvae and Ascosphaera apis. A chemical characterization of kefir was also carried out. Our results confirmed that bacteria and yeasts were more numerous in kefir grains compared with those in the beverage. The counts of microorganisms declined, although an important microbial community was still present in kefir after the long storage period. Eleven strains which inhibited bee pathogens were isolated from kefir. Genotypic results demonstrated that these isolates included Lentilactobacillus hilgardii, Lentilactobacillus buchneri and Saccharomyces cerevisiae. Thus, water kefir may be an innovative source of potential probiotic strains for bee nutrition in order to control honeybee diseases.

Ganoderma sessile is a fast polysaccharide producer among Ganoderma species.

The selection of fast-growing and high-yield-producing strains is required to satisfy the market demand on fungal food supplements. To that aim, three strains deposited in our collection as G. lucidum and G. oregonense were screened for polysaccharide production and biomass yield. Ganoderma strains deposited as G. lucidum were identified as G. sessile and G. lingzhi by nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 (ITS) and translation elongation factor 1-α (TEF1-α) phylogenies. The identity of G. oregonense was confirmed by molecular phylogeny and biogeography. Additionally, mycelial antagonism confirmed species differentiation, and strains were further distinguished by morphology and protein profiles. Biomass and polysaccharide yields of G. sessile were clearly different from those of G. lingzhi and G. oregonense in both liquid culture and solid-state fermentation. The maximum polysaccharide yield (4.52 ± 0.83 g L-1) for G. sessile was obtained from submerged cultures at day 9. G. sessile also achieved the highest linear growth in lignocellulosic solid substrates. Consequently, basidiomata were successfully obtained by solid-state fermentation in polypropylene bags, whereas G. lingzhi and G. oregonense mushrooms were not produced in artificial solid substrates. G. sessile, a species frequently collected in America, showed to be a promising polysaccharide producer for the manufacture of dietary supplements.

Transcriptomic response of durum wheat to cold stress at reproductive stage.

Understanding the genetic basis of cold tolerance is a key step towards obtaining new and improved crop varieties. Current geographical distribution of durum wheat in Argentina exposes the plants to frost damage when spikes have already emerged. Biochemical pathways involved in cold tolerance are known to be early activated at above freezing temperatures. In this study we reported the transcriptome of CBW0101 spring durum wheat by merging data from untreated control and cold (5 °C) treated plant samples at reproductive stage. A total of 128,804 unigenes were predicted. Near 62% of the unigenes were annotated in at least one database. In total 876 unigenes were differentially expressed (DEGs), 562 were up-regulated and 314 down-regulated in treated samples. DEGs are involved in many critical processes including, photosynthetic activity, lipid and carbohydrate synthesis and accumulation of amino acids and seed proteins. Twenty-eight transcription factors (TFs) belonging to 14 families resulted differentially expressed from which eight families comprised of only TFs induced by cold. We also found 31 differentially expressed Long non-coding RNAs (lncRNAs), most of them up-regulated in treated plants. Two of these lncRNAs could operate via microRNAs (miRNAs) target mimic. Our results suggest a reprogramming of expression patterns in CBW0101 that affects a number of genes that is closer to the number reported in winter genotypes. These observations could partially explain its moderate tolerance (low proportion of frost-damaged spikes) when exposed to freezing days in the field.

Exploring the life cycles of three South American rusts that have potential as biocontrol agents of the stipoid grass Nassella neesiana in Australasia.

Three rusts, Puccinia nassellae, Uromyces pencanus, and Puccinia graminella, are being studied as potential biological control agents for Nassella neesiana (Chilean needle grass) in Australia and New Zealand. An understanding of the life cycle of a pathogen is desirable before its release as a biocontrol agent is considered, to enable the assessment of the risks involved in such a release. Field observations and experiments have been carried out to elucidate the life cycles of these three pathogens. Puccinia nassellae cycles as urediniospores and produces dormant teliospores. Dormancy of teliospores has been broken through manipulation in the laboratory, but resulting basidiospores have failed to infect N. neesiana plants under the conditions tested. Uromyces pencanus cycles as urediniospores and its telia appear to have lost the capacity to produce basidiospores. Aecia have been reported for this rust in the literature. However, evidence is provided that these aecia in fact belong to the life cycle of P. graminella. Puccinia graminella produces only aecia and telia. The aeciospores have been shown to be repetitive (aecidioid urediniospores). Teliospores germinate directly without a dormant phase, but resulting basidiospores failed to infect N. neesiana plants under the conditions tested. The role of teliospores in the life cycle of all three rusts remains unknown. Although the autoecious nature of their life cycles has not been proven experimentally, neither is there any evidence that they are heteroecious. Practical and theoretical implications of these findings are discussed.