The nutritive value of Rhodes grass (Chloris gayana) when treated with CaO, NaOH or a microbial inoculant and offered to dairy heifers as big-bale silage

A series of laboratory and animal studies examined the use of chemical and biological agents to enhance the digestibility of Rhodes grass (grass) cut at 60 (young) and 100 (mature) days of regrowth and ensiled as big round bales. The treatments included an untreated control (C), a microbial inoculant (I), NaOH, CaO and NaOH plus inoculant (NaOH + I). Inoculant was grown anaerobically, using a starter culture of rumen fluid from cattle given Rhodes grass. Treatments C, 1, NaOH, NaOH + I, were offered separately to twelve dairy heifers, in a 3 X 4 randomized complete block design, repeated twice for each grass silage. C and I had substantial mould growth, compared with no visible mould in NaOH or NaOH + 1. CaO treatment was effective in preventing mould growth, but had little effect on the chemical composition and in sacco digestibility of mature grass silage. NaOH reduced NDF content and increased in sacco digestibility (P < 0.05) but not the in vivo digestibility (P > 0.05) of both mature- and young-grass silage. The effects of other treatments on nutritive value were non-significant at both stages of maturity. NaOH increased the intake of mature-grass silage by 24-26% (P < 0.05), but had little effect on the intake of young-grass silage (P > 0.05). Treatment I consistently reduced grass silage intake (P < 005) for young-grass silage. The findings of these studies show that treating mature Rhodes grass with NaOH will improve its nutritive value and reduce mould growth in conserved herbage. However none of the treatments in this study had any consistently positive effects on the in vivo nutritive value or storage quality of young-grass silage.

PLANT GROWTH-PROMOTING MICROBIAL INOCULANT FOR Schizolobium parahyba pv. parahyba

ABSTRACTSchizolobium parahyba pv. amazonicum (Huber ex Ducke) Barneby (paricá) occurs naturally in the Amazon and is significant commercial importance due to its rapid growth and excellent performance on cropping systems. The aim of this paper was to evaluate a microbial inoculants such as arbuscular mycorrhiza fungi (AMF) and Rhizobium sp. that promote plant growth. The inocula was 10 g of root colonized and spores of Glomus clarum and/or 1 mL of cell suspension (107 CFU/mL) of Rhizobium sp. and/or 100 g of chemical fertilizer NPK 20-05-20 per planting hole. The experimental design was complete randomized blocks with five replications and eight treatments (n = 800). Plant height, stem diameter and plant survival were measured. The results were tested for normality and homogeneity of variances and analyzed by ANOVA and Tukey test (p < 0.05). Rhizobium sp and AM fungi showed no effect on plant growth. Environmental factors probably influenced the effectiveness of symbiosis of both microorganisms and plant growth. The chemical fertilizer increased S. parahyba growth. During the first 120 days plants suffered with drought and frost, and at 180 days plants inoculated with microorganism plus chemical fertilizer showed higher survival when compared with control. The results showed that the microbial inoculants used showed an important role on plant survival after high stress conditions, but not in plant growth. Also was concluded that the planting time should be between November to December to avoid the presence of young plants during winter time that is dry and cold.

Estabilidade aeróbica de silagens de capim-elefante (Pennisetum purpureum, Schum) emurchecido e tratado com inoculante microbiano

Para avaliar o efeito de um inoculante microbiano sobre a estabilidade aeróbica de silagens de capim-elefante (Pennisetum purpureum, Schum) pré-seco, foi conduzido um experimento na UNESP, Campus de Jaboticabal. O capim foi cortado com 96 dias de crescimento, permanecendo sob ação do sol, por aproximadamente 5 horas para o emurchecimento. Foram confeccionados silagens, em barricas plásticas de 200 L de capacidade, que receberam (CI) ou não (SI) o inoculante no momento da ensilagem. Para o estudo da deterioração aeróbica, na abertura dos silos (T0), cada barrica foi subdividida em três profundidades e as silagens expostas ao ar em um sistema, usado para a determinação de CO2, por 2 (T2), 4 (T4), 6 (T6) e 8 (T8) dias. A silagem da camada superficial não apresentou diferenças no valor de pH e contagem de leveduras, em relação à da camada intermediária, porém ambas apresentaram valores superiores à camada mais profunda. O teor de N-NH3 da camada superior foi menor que o da intermediária e profunda, que não diferiram entre si. A interação significativa entre inóculo e tempo de exposição mostrou que nos primeiros tempos de aeração (T0; T2 e T4) as silagens inoculadas não diferiram das controle em termos de pH, teor de N-NH3 e contagem de bolores, mas com avanço do tempo (T6 e T8) passaram a apresentar menor valor de pH, menor contagem de fungos e tendência a menor produção de CO2, indicando menor velocidade de deterioração após seis dias de exposição aeróbica.

Estabilidade aeróbia de silagens de planta e de grãos úmidos de milho

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bioremediation of Herbicide Velpar K (R) In Vitro in Aqueous Solution with Application of EM-4 (Effective Microorganisms)

This work assessed the bioremediation of herbicide Velpar K (R), in vitro in aqueous solution, used against weeds in sugar cane in Sao Paulo state. The herbicide contained Hexazinone and Diuron. It was used the microbial inoculant denominated Effective Microorganisms (EM-4), pool of microorganisms from soil that contained lactic and photosynthetic bacteria, fungi, yeasts and actinomycetes for bioremediation. Results for the depth of cultivation on agar-agar inoculated with EM-4 showed the microorganisms growth in the concentrations between 0.2% and 1.0% of the Velpar K (R) in the gel. The analysis of high performance liquid chromatography ( HPLC) showed that the EM-4 was effective for the bioremediation of the herbicide, which reached the values of 80% for diuron and 70% for hexazinone after 21 days in solution of 2:1 of Velpar K (R)/EM-4 ratio. These results could be useful for planning the bioremediation of contaminated areas with Velpar K (R).

Bioremediation of herbicide velpar K® in vitro in aqueous solution with application of EM-4 (effective microorganisms)

This work assessed the bioremediation of herbicide Velpar K®, in vitro in aqueous solution, used against weeds in sugar cane in São Paulo state. The herbicide contained Hexazinone and Diuron. It was used the microbial inoculant denominated Effective Microorganisms (EM-4), pool of microorganisms from soil that contained lactic and photosynthetic bacteria, fungi, yeasts and actinomycetes for bioremediation. Results for the depth of cultivation on agar-agar inoculated with EM-4 showed the microorganisms growth in the concentrations between 0.2% and 1.0% of the Velpar K®in the gel. The analysis of high performance liquid chromatography (HPLC) showed that the EM-4 was effective for the bioremediation of the herbicide, which reached the values of 80% for diuron and 70% for hexazinone after 21 days in solution of 2:1 of Velpar K®/EM-4 ratio. These results could be useful for planning the bioremediation of contaminated areas with Velpar K®.

Persistence of a biocontrol Pseudomonas inoculant as high populations of culturable and non-culturable cells in 200-cm-deep soil profiles

Little is known about the ecology of soil inoculants used for pathogen biocontrol, biofertilization and bioremediation under field conditions. We investigated the persistence and the physiological states of soil-inoculated Pseudomonas protegens (previously Pseudomonas fluorescens) CHA0 (108 CFU g−1 surface soil) in different soil microbial habitats in a planted ley (Medicago sativa L.) and an uncovered field plot. At 72 days, colony counts of the inoculant were low in surface soil (uncovered plot) and earthworm guts (ley plot), whereas soil above the plow pan (uncovered plot), and the rhizosphere and worm burrows present until 1.2 m depth (ley plot) were survival hot spots (105-106 CFU g−1 soil). Interestingly, strain CHA0 was also detected in the subsoil of both plots, at 102-105 CFU g−1 soil between 1.8 and 2 m depth. However, non-cultured CHA0 cells were also evidenced based on immunofluorescence microscopy. Kogure's direct viable counts of nutrient-responsive cells showed that many more CHA0 cells were in a viable but non-culturable (VBNC) or a non-responsive (dormant) state than in a culturable state, and the proportion of cells in those non-cultured states depended on soil microbial habitat. At the most, cells in a VBNC state amounted to 34% (above the plow pan) and those in a dormant state to 89% (in bulk soil between 0.6 and 2 m) of all CHA0 cells. The results indicate that field-released Pseudomonas inoculants may persist at high cell numbers, even in deeper soil layers, and display a combination of different physiological states whose prevalence fluctuates according to soil microbial habitats.

Corn silage inoculated with microbial additives

Pós-graduação em Zootecnia - FCAV

Microbial Evaluation Of Traumatized Teeth Treated With Triple Antibiotic Paste Or Calcium Hydroxide With 2% Chlorhexidine Gel In Pulp Revascularization.

Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments. Fifteen patients (7-17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA). Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features. The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.

Phylogenetic Analysis Of The Microbial Community In Hypersaline Petroleum Produced Water From The Campos Basin.

In this work the archaea and eubacteria community of a hypersaline produced water from the Campos Basin that had been transported and discharged to an onshore storage facility was evaluated by 16S recombinant RNA (rRNA) gene sequence analysis. The produced water had a hypersaline salt content of 10 (w/v), had a carbon oxygen demand (COD) of 4,300 mg/l and contains phenol and other aromatic compounds. The high salt and COD content and the presence of toxic phenolic compounds present a problem for conventional discharge to open seawater. In previous studies, we demonstrated that the COD and phenolic content could be largely removed under aerobic conditions, without dilution, by either addition of phenol degrading Haloarchaea or the addition of nutrients alone. In this study our goal was to characterize the microbial community to gain further insight into the persistence of reservoir community members in the produced water and the potential for bioremediation of COD and toxic contaminants. Members of the archaea community were consistent with previously identified communities from mesothermic reservoirs. All identified archaea were located within the phylum Euryarchaeota, with 98 % being identified as methanogens while 2 % could not be affiliated with any known genus. Of the identified archaea, 37 % were identified as members of the strictly carbon-dioxide-reducing genus Methanoplanus and 59 % as members of the acetoclastic genus Methanosaeta. No Haloarchaea were detected, consistent with the need to add these organisms for COD and aromatic removal. Marinobacter and Halomonas dominated the eubacterial community. The presence of these genera is consistent with the ability to stimulate COD and aromatic removal with nutrient addition. In addition, anaerobic members of the phyla Thermotogae, Firmicutes, and unclassified eubacteria were identified and may represent reservoir organisms associated with the conversion hydrocarbons to methane.

Microbial Lipid Production: Screening With Yeasts Grown On Brazilian Molasses.

Rhodotorula glutinis CCT 2182, Rhodosporidium toruloides CCT 0783, Rhodotorula minuta CCT 1751 and Lipomyces starkeyi DSM 70296 were evaluated for the conversion of sugars from Brazilian molasses into single-cell oil (SCO) feedstock for biodiesel. Pulsed fed-batch fermentations were performed in 1.65 l working volume bioreactors. The maximum specific growth rate (µmax), lipid productivity (Pr) and cellular lipid content were, respectively, 0.23 h(-1), 0.41 g l(-1) h(-1), and 41% for Rsp. toruloides; 0.20 h(-1), 0.27 g l(-1) h(-1), and 36% for Rta. glutinis; 0.115 h(-1), 0.135 g l(-1) h(-1), and 27 % for Rta. minuta; and 0.11 h(-1), 0.13 g l(-1) h(-1), and 32% for L. starkeyi. Based on their microbial lipid productivity, content, and profile, Rsp. toruloides and Rta. glutinis are promising candidates for biodiesel production from Brazilian molasses. All the oils from the yeasts were similar to the composition of plant oils (rapeseed and soybean) and could be used as raw material for biofuels, as well as in food and nutraceutical products.

Comparative Metagenomic Analysis Of Coral Microbial Communities Using A Reference-independent Approach.

By comparing the SEED and Pfam functional profiles of metagenomes of two Brazilian coral species with 29 datasets that are publicly available, we were able to identify some functions, such as protein secretion systems, that are overrepresented in the metagenomes of corals and may play a role in the establishment and maintenance of bacteria-coral associations. However, only a small percentage of the reads of these metagenomes could be annotated by these reference databases, which may lead to a strong bias in the comparative studies. For this reason, we have searched for identical sequences (99% of nucleotide identity) among these metagenomes in order to perform a reference-independent comparative analysis, and we were able to identify groups of microbial communities that may be under similar selective pressures. The identification of sequences shared among the metagenomes was found to be even better for the identification of groups of communities with similar niche requirements than the traditional analysis of functional profiles. This approach is not only helpful for the investigation of similarities between microbial communities with high proportion of unknown reads, but also enables an indirect overview of gene exchange between communities.

Microbial distribution in the root canal system after periapical lesion induction using different methods

The aim of this study was to evaluate the microbial distribution in the root canal system after periapical lesion induction in dogs' teeth using different methods. Fifty-two root canals were assigned to 4 groups (n=13). Groups I and II: root canals were exposed to the oral cavity for 180 days; groups III and IV: root canals were exposed for 7 days and then the coronal openings were sealed for 53 days. The root apices of groups I and III were perforated, while those of groups II and IV remained intact. After the experimental periods, the animals were euthanized and the anatomic pieces containing the roots were processed and stained with the Brown & Brenn method to assess the presence and distribution of microorganisms. The incidence of microorganisms at different sites of the roots and periapical lesions was analyzed statistically by the chi-square test at 5% significance level. All groups presented microorganisms in the entire root canal system. A larger number of microorganisms was observed on the root canal walls, apical delta and dentinal tubules (p<0.05), followed by cementum and cemental resorption areas. In spite of the different periods of exposure to the oral environment, the methods used for induction of periapical periodontitis yielded similar distribution of microorganisms in the root canal system.

Anticariogenic Properties of ent-Pimarane Diterpenes Obtained by Microbial Transformation

In the present work, the anticariogenic activities of three pimarane-type diterpenes obtained by fungal biotransformation were investigated. Among these metabolites, ent-8(14), 15-pimaradien-19-ol was the most active compound, displaying very promising MIC values (ranging from 1.5 to 4.0 mu g mL(-1)) against the main microorganisms responsible for dental caries: Streptococcus salivarius, S. sobrinus, S. mutans, S. mitis, S. sanguinis, and Lactobacillus casei. Time kill assays performed with ent-8(14), 15-pimaradien-19-ol against the primary causative agent S. mutans revealed that this compound only avoids growth of the inoculum in the first 12 h (bacteriostatic effect). However, its bactericidal effect is clearly noted thereafter (between 12 and 24 h). The curve profile obtained by combining ent-8(14), 15-pimaradien-19-ol and chlorhexidine revealed a significant reduction in the time necessary for killing S. mutans compared with each of these two chemicals alone. However, no synergistic effect was observed using the same combination in the checkerboard assays against this microorganism. In conclusion, our results point out that ent-8(14), 15-pimaradien-19-ol is an important metabolite in the search for new effective anticariogenic agents.