Actinoplanes flavus sp. nov., a novel cellulase-producing actinobacterium isolated from coconut palm rhizosphere soil.

A novel cellulase-producing actinomycete, designated strain NEAU-H7T, was isolated from coconut palm rhizosphere soil collected from Wenchang City, Hainan Province, PR China. A polyphasic taxonomic study was carried out to establish the status of this strain. Results of 16S rRNA gene sequence analysis indicated that strain NEAU-H7T belonged to the genus Actinoplanes, with highest similarity to Actinoplanes hulinensis NEAU-M9T (99.2 % 16S rRNA gene sequence similarity). The diagnostic sugars in cell hydrolysates were determined to be ribose, galactose and mannose. The major fatty acids (>10%) were C16 : 0, C18 : 1 ω9c and C18 : 0. The predominant menaquinones were identified as MK-9(H4) and MK-9(H6). The major polar lipids were phosphatidylethanolamine, phosphatidylinositol and two phosphatidylinositol mannosides. The amino acid of the cell-wall peptidoglycan was determined to be meso-diaminopimelic acid. The DNA G+C content was 71.2 mol%. Phylogenetic analysis using 16S rRNA gene sequences showed that strain NEAU-H7T formed a stable phyletic line with A. hulinensis NEAU-M9T. However, whole-genome phylogeny showed strain NEAU-H7T formed a stable phyletic line with A. hulinensis NEAU-M9T (99.2%), Actinoplanes campanulatus DSM 43148T (98.6%), Actinoplanes capillaceus DSM 44859T (98.3%) and Actinoplanes lobatus DSM 43150T (97.6%). The digital DNA-DNA hybridization (dDDH) results between them were 53.6 (50.9-56.2), 54.1 (51.3-56.9), 53.1 (50.3-55.9) and 52.9 % (50.1-55.6 %), and whole-genome average nucleotide identity (ANI) values between them were 93.7, 93.6, 93.5 and 93.5 %. The low dDDH and ANI values demonstrated that strain NEAU-H7T could be distinguished from its reference strains. Moreover, genomic analysis indicated that the strain NEAU-H7T had the potential to decompose cellulose and produce bioactive compounds. On the basis of morphological, chemotaxonomic and phylogenetic characteristics, strain NEAU-H7T is proposed to represent a novel species of the genus Actinoplanes, with the name Actinoplanes flavus sp. nov. The type strain is NEAU-H7T (=CCTCC AA 2020034T=DSM 112042T).

The use of High-Pressure Processing (HPP) to improve the safety and quality of raw coconut (Cocos nucifera L) water.

This research investigated the use of high-pressure processing (HPP) for inactivating vegetative pathogens and spoilage microbiota in fresh unfiltered coconut water (Cocos nucifera L) from nuts obtained from Florida and frozen CW from Brazil with pH >5.0 and storage at 4 °C. Additionally, CW was evaluated to determine if it supported the growth and toxin production of Clostridium botulinum with or without the use of HPP when stored at refrigeration temperatures. Samples of fresh unfiltered CW were inoculated to 5.5 to 6.5 logs/mL with multiple strain cocktails of E. coli O157:H7, Salmonella spp. and Listeria monocytogenes and HPP at 593 MPa for 3 min at 4 °C. HPP and inoculated non-HPP controls were stored at 4 °C for 54 and 75 days for Florida CW and Brazil CW, respectively. Results of analyses showed HPP samples with <1 CFU/mL and no detection (negative/25 mL) with enrichment procedures for the 3 inoculated pathogens for all analyses. The non-HPP control samples did not show growth of the pathogens but a gradual decrease in levels to ca. 3-Logs/mL by day 54 in the fresh Florida CW and similarly in frozen Brazil CW by Day 75. Microbial spoilage of uninoculated samples was evaluated for normal spoilage microbiota through 120 days storage at 4 °C. Microbial counts remained at ca. 2-logs with no detectable signs of spoilage for HPP samples through 120 d. The non-HPP control samples spoiled within 2 weeks of storage at 4 °C with gas production, cloudiness, and off-odors. To evaluate if CW supports the growth and toxin production of C. botulinum, samples of unfiltered and filtered (0.2 µm) CW were inoculated with either proteolytic or non-proteolytic C. botulinum spores at 2 log CFU/mL that were processed at 593 MPa for 3 min and stored at 4 °C and 10 °C for 45 days. Inoculated positive and non-inoculated negative controls were prepared and stored as the HPP treated and non-HPP samples. No growth of C. botulinum or toxin production was detected in either the unfiltered or filtered CW regardless if products were HPP treated or not. All inoculated samples with C. botulinum spores were enriched at Day-45 in PYGS media to determine the viability of the inoculated spores at the end of shelf-life and screened for C. botulinum toxins. In all samples, C. botulinum toxin Types A, B and E were detected indicating spores were viable throughout the storage. Type F toxin was not detected possibly due to inherent conditions in the samples that may affected toxin screening.

Inactivation of Bacillus and Clostridium Spores in Coconut Water by Ultraviolet Light.

Bacterial spores are generally more resistant than vegetative bacteria to ultraviolet (UV) inactivation. The UV sensitivity of these spores must be known for implementing UV disinfection of low acid liquid foods. UV inactivation kinetics of bacterial spores in coconut water (CW) and distilled sterile water was studied. Populations of Bacillus cereus and Clostridium sporogenes dormant spores were reduced by more than 5.5 log10 at the UV-C photon fluence of 1142 µE·m-2 and 1919 µE·m-2 respectively. C. sporogenes spores showed higher UV-C resistance than B. cereus, with the photon fluence 300 µE·m-2 required for one log inactivation (D10) and 194 µE·m-2, respectively. No significant difference was observed in D10 values of spores suspended in the two fluid types (p > 0.05). The inactivation kinetics of microorganisms were described by log linear models with low root mean square error and high coefficient of determination (R2 > 0.98). This study clearly demonstrated that high levels of inactivation of bacterial spores can be achieved in CW. The baseline data generated from this study will be used to conduct spore inactivation studies in continuous flow UV systems. Further proliferation of the technology will include conducting extensive pilot studies.

Individual and combined efficacies of mild heat and ultraviolet-c radiation against Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in coconut liquid endosperm.

This study determined the inactivation kinetic parameters of selected pathogens in heat, ultraviolet-C and combined heat-UV-C treated coconut liquid endosperm. Separate cocktails of Escherichia coli O157:H7, Salmonella enterica serovars, and Listeria monocytogenes strains were inoculated into coconut liquid endosperm (pH 5.15, TSS 4.4oBx, TA 0.062% malic acid, extinction coefficient (ε) at 254 nm of 0.0154 cm-1) for inactivation studies. Result showed that all organisms generally exhibited a log-linear heat inactivation behavior (R2 0.81-0.99). The E. coli O157:H7 cocktail (D55 = 19.75 min, D57 = 10.79 min, D60 = 3.38 min, and D63 = 0.46 min) was found to be significantly more resistant (P > 0.05) than the tested cocktail of L. monocytogenes (D55 = 11.68 min, D57 = 4.53 min, D60 = 1.82 min and D63 = 0.26 min) and S. enterica cocktail (D55 = 3.08 min, D57 = 2.60 min, D60 = 0.89 min and D63 = 0.25 min). Despite the differences in DT values, computed z values for L. monocytogenes cocktail (5.12 ±â€¯0.43 °C) and E. coli O157:H7 cocktail (4.95 ±â€¯0.12 °C) were not significantly different (P > 0.05), but were both significantly (P < 0.05) lower than that of S. enterica cocktail (7.10 ±â€¯0.15 °C). All test organisms also exhibited a generally log-linear UV-C inactivation behavior (R2 0.90-0.99) with E. coli O157:H7 cocktail (DUV-C = 25.26 mJ/cm2) demonstrating greatest resistance to UV-C than S. enterica (DUV-C = 24.65 mJ/cm2) and L. monocytogenes (DUV-C = 17.30 mJ/cm2) cocktails. The D55 values of each organism cocktail were used to calculate for the 3-log reduction heating process schedules, during which UV-C treatments were simultaneously applied. Lethal rates (F values) calculations in the combined processes revealed that within the 3-log reduction heating processes, co-exposure of UV-C resulted in 5.62 to 6.20 log reductions in the test organism populations. Heating caused 69.3, 97.2, and 67.4% of the reduction in E. coli O157:H7, S. enterica and L. monocytogenes cocktails, respectively. These results can be used as baseline data in the establishment of mild heat treatment in combination with UV-C process schedules for coconut liquid endosperm and other similar products.

Polyphasic characterization of a novel species in the Lactobacillus casei group from cow manure of Taiwan: Description of L. chiayiensis sp. nov.

Two Gram-stain-positive, rod-shaped, non-motile, catalase-negative and facultative anaerobic strains, NCYUAST and BCRC 18859 (=NRIC 1947), were isolated from cow manure of Taiwan and coconut juice of Philippines, respectively. Comparative sequence analysis of 16S rRNA gene revealed that the novel strains were members of the genus Lactobacillus. These two strains had 100% of 16S rRNA gene sequence similarity and 98.6% of average nucleotide identity (ANI) value based on whole genome sequences. On the basis of 16S rRNA gene sequence similarity, the type strains of Lactobacillus casei (99.6% similarity), Lactobacillus paracasei subsp. paracasei (99.1%), L. paracasei subsp. tolerans (99.1%), Lactobacillus rhmnosus (99.0%) and 'Lactobacillus zeae' (99.7%) were the closest neighbors to these novel strains. The results of phenotypic and chemotaxonomic characterization, multilocus sequence analysis (MLSA) based on the sequences of three housekeeping genes (dnaK, pheS and yycH), whole-genome sequence (WGS)-based comparison by ANI and in silico DNA-DNA hybridization (isDDH), species-specific PCR and whole-cell MALDI-TOF MS spectral pattern analyses demonstrated that the novel two strains represented a single, novel species within the L. casei group, for which the name Lactobacillus chiayiensis sp. nov., is proposed. The type strain is NCYUAST (=BCRC 81062T=NBRC 112906T).

Microbial inactivation and cytotoxicity evaluation of UV irradiated coconut water in a novel continuous flow spiral reactor.

A continuous-flow UV reactor operating at 254nm wave-length was used to investigate inactivation of microorganisms including bacteriophage in coconut water, a highly opaque liquid food. UV-C inactivation kinetics of two surrogate viruses (MS2, T1UV) and three bacteria (E. coli ATCC 25922, Salmonella Typhimurium ATCC 13311, Listeria monocytogenes ATCC 19115) in buffer and coconut water were investigated (D10 values ranging from 2.82 to 4.54mJ·cm-2). A series of known UV-C doses were delivered to the samples. Inactivation levels of all organisms were linearly proportional to UV-C dose (r2>0.97). At the highest dose of 30mJ·cm-2, the three pathogenic organisms were inactivated by >5 log10 (p<0.05). Results clearly demonstrated that UV-C irradiation effectively inactivated bacteriophage and pathogenic microbes in coconut water. The inactivation kinetics of microorganisms were best described by log linear model with a low root mean square error (RMSE) and high coefficient of determination (r2>0.97). Models for predicting log reduction as a function of UV-C irradiation dose were found to be significant (p<0.05) with low RMSE and high r2. The irradiated coconut water showed no cytotoxic effects on normal human intestinal cells and normal mouse liver cells. Overall, these results indicated that UV-C treatment did not generate cytotoxic compounds in the coconut water. This study clearly demonstrated that high levels of inactivation of pathogens can be achieved in coconut water, and suggested potential method for UV-C treatment of other liquid foods. INDUSTRIAL RELEVANCE: This research paper provides scientific evidence of the potential benefits of UV-C irradiation in inactivating bacterial and viral surrogates at commercially relevant doses of 0-120mJ·cm-2. The irradiated coconut water showed no cytotoxic effects on normal intestinal and healthy mice liver cells. UV-C irradiation is an attractive food preservation technology and offers opportunities for horticultural and food processing industries to meet the growing demand from consumers for healthier and safe food products. This study would provide technical support for commercialization of UV-C treatment of beverages.

Comparison of in vivo toxicity, antioxidant and immunomodulatory activities of coconut, nipah and pineapple juice vinegars.

BACKGROUND: Vinegar is widely used as a food additive, in food preparation and as a food supplement. This study compared the phenolic acid profiles and in vivo toxicities, and antioxidant and immunomodulatory effects of coconut, nipah and pineapple juice vinegars, which were respectively prepared via a two-step fermentation using Saccharomyces cerevisiae 7013 INRA and Acetobacter aceti vat Europeans. RESULTS: Pineapple juice vinegar, which had the highest total phenolic acid content, also exhibited the greatest in vitro antioxidant capacity compared to coconut juice and nipah juice vinegars. Following acute and sub-chronic in vivo toxicity evaluation, no toxicity and mortality were evident and there were no significant differences in the serum biochemical profiles between mice administered the vinegars versus the control group. In the sub-chronic toxicity evaluation, the highest liver antioxidant levels were found in mice fed with pineapple juice vinegar, followed by coconut juice and nipah juice vinegars. However, compared to the pineapple juice and nipah juice vinegars, the mice fed with coconut juice vinegar, exhibited a higher population of CD4+ and CD8+ T-lymphocytes in the spleen, which was associated with greater levels of serum interleukin-2 and interferon-γ cytokines. CONCLUSIONS: Overall, the data suggested that not all vinegar samples cause acute and sub-chronic toxicity in vivo. Moreover, the in vivo immunity and organ antioxidant levels were enhanced, to varying extents, by the phenolic acids present in the vinegars. The results obtained in this study provide appropriate guidelines for further in vivo bioactivity studies and pre-clinical assessments of vinegar consumption. © 2017 Society of Chemical Industry.

Physicochemical profile of microbial-assisted composting on empty fruit bunches of oil palm trees.

This study was carried out to investigate the physicochemical properties of compost from oil palm empty fruit bunches (EFB) inoculated with effective microorganisms (EM∙1™). The duration of microbial-assisted composting was shorter (∼7 days) than control samples (2 months) in a laboratory scale (2 kg) experiment. The temperature profile of EFB compost fluctuated between 26 and 52 °C without the presence of consistent thermophilic phase. The pH of compost changed from weak acidic (pH ∼5) to mild alkaline (pH ∼8) because of the formation of nitrogenous ions such as ammonium (NH4 (+)), nitrite (NO2 (-)), and nitrate (NO3 (-)) from organic substances during mineralization. The pH of the microbial-treated compost was less than 8.5 which is important to prevent the loss of nitrogen as ammonia gas in a strong alkaline condition. Similarly, carbon mineralization could be determined by measuring CO2 emission. The microbial-treated compost could maintain longer period (∼13 days) of high CO2 emission resulted from high microbial activity and reached the threshold value (120 mg CO2-C kg(-1) day(-1)) for compost maturity earlier (7 days). Microbial-treated compost slightly improved the content of minerals such as Mg, K, Ca, and B, as well as key metabolite, 5-aminolevulinic acid for plant growth at the maturity stage of compost. Graphical Abstract Microbial-assisted composting on empty fruit bunches.

Eco-epidemiologic study of emerging fungi related to the work of babacu coconut breakers in the State of Maranhao, Brazil

Introduction: There are more than 300,000 extractors using the babaçu coconut as a source of income in the States of Maranhão, Pará, Tocantins and Piauí, and this activity is associated with fungal infections. The objective of this study was to examine the occurrence of emergent fungi in the conjunctiva, nails and surface and subcutaneous injuries of female coconut breakers in Esperantinópolis, Maranhão. Additionally, soil samples and palm structures were collected. Methods: The obtained samples were cultured in Petri dishes containing potato-dextrose-agar and chloramphenicol. The etiological agent was confirmed by a direct mycological exam and growth in culture. Results: In total, 150 domiciles were visited, and samples were collected from 80 patients. From the ground, the most frequently isolated fungus was Aspergillus niger (53. 8%). the most frequently detected fungus in babaçu coconut was Aspergillus niger (66.7%). Conjunctival fungal growth occurred in 76.3% of the women. The ocular fungal microbiota consisted of filamentous fungi (80.6%), and yeasts were present in 19.4% of cases. Onychomycosis was diagnosed in 44% (11/25) of the women. Conclusions: The identification of the genera Neosartorya, Rhizopus and Curvularia in onychomycoses shows that emergent filamentous fungi can be isolated. Aspergillus sp., Penicillium sp. and Scedosporium sp. were the predominant genera found in the babaçu coconut. From ocular conjunctiva, Candida spp. were the most prevalent species isolated, and Fusarium sp. was present only in one woman. The nearly permanent exposure of coconut breakers to the external environment and to the soil is most likely the reason for the existence of a mycotic flora and fungal infections, varying according to the individual's practices and occupation. .

Caracterização química de águas de coco de três variedades / Characterization chemical of coconut water for three varieties

O presente trabalho teve como objetivo avaliar as características físicas e químicas do coco e das águas de coco de três variedades: Anão Verde, Híbrido e Anão Amarelo. Nas determinações: peso do fruto, comprimento longitudinal e transversal, volume da água de coco, rendimento da polpa, sólidos solúveis totais, acidez total titulável, pH e condutividade elétrica, apenas esta última apresentou diferença estatisticamente significante (p=0,05) entre as variedades estudadas. Na análise dos resultados, no que diz respeito às características físicas, a variedade Híbrido apresentou maior peso e volume de água, enquanto a Anão Verde revelou maior rendimento de polpa. Em relação aos açúcares redutores totais e sacarose, a variedade Híbrido apresentou maiores teores que Anão Amarelo e Anão Verde (p 0,05). Dentre os minerais (Na, K, Ca, P e Mg), apenas o sódio apresentou diferença significante (p 0,05) entre as variedades pesquisadas.

Avaliaçäo da eficiência de diferentes tratamentos térmicos na estabilidade microbiológica do leite de coco / Evaluation of the effectiveness of different heating treatments on the microbiological stability of coconut milk

As indústrias de leite de coco no Brasil utilizam como método de conservaçäo deste produto a pasteurizaçäo acompanhada da adiçäo de conservadores químicos do tipo metabissulfito de sódio, sorbato de potássio e benzoato de sódio. Devido às características do produto (pH 6,0 e atividade de água maior que 0,85), esse tratamento näo é efetivo para garantir sua estabilidade microbiológica. Estudou-se entäo a estabilidade microbiológica do leite de coco, comercializado à temperatura ambiente, através da avaliaçäo da eficiência dos processos termoquímicos mais comumente empregados pelas indústrias, comparando-os ao processo de esterilizaçäo em autocalve rotativa. O leite de coco foi previamente formulado e acrescido de esporos de Clostridium sporogenes (PA 3679). Para cada processo utilizaram-se dois níveis de inoculaçäo, um de 2x dez ao quadrado e outro de 2x dez a sexta esporos/200ml. Os resultados obtidos demonstraram que existe a possibilidade do desenvolvimento de clostrídios anaeróbicos putrefativos, indicando inclusive o risco de desenvolvimento de Clostridium botulinum quando näo se utiliza a esterilizaçäo do leite de coco em autoclave rotativa. Portanto, o tratamento térmico à pressäo atmosférica praticado pela indústria implica em risco potencial à saúde pública