Spencerozyma pingqiaoensis sp. nov., a yeast species isolated from the external surface of rice leaves in China.

Four strains (NYNU 178247, NYNU 178251, DMKU-PAL160 and DMKU-PAL137) representing a novel yeast species were isolated from the external surfaces of rice and pineapple leaves collected in China and Thailand. Phylogenetic analysis based on the concatenated sequences of the internal transcribed spacer (ITS) regions and the D1/D2 domains of the large subunit rRNA gene revealed that the novel species belonged to the genus Spencerozyma. The D1/D2 sequence of the novel species differed from its closest relative, Spencerozyma acididurans SYSU-17T, by 3.2 % sequence divergence. The species also differed from Spencerozyma crocea CBS 2029T and Spencerozyma siamensis DMKU13-2T, by 3.0-6.9 % sequence divergence in the D1/D2 sequences out of 592 bp. In the ITS regions, the novel species displayed 19.8-29.2% sequence divergence from S. acididurans SYSU-17T, S. crocea CBS 2029T and S. siamensis DMKU13-2T out of 655 bp. Furthermore, the novel species could also be differentiated from the closely related species by some physiological characteristics. The species name of Spencerozyma pingqiaoensis sp. nov. (Holotype CBS 15238, Mycobank MB 844734) is proposed to accommodate these four strains.

Impact of Pineapple Juice on Expression of CYP3A4, NAT2, SULT1A1 and OATP1B1 mRNA in HepG2 Cells.

<b>Background and Objective:</b> Pineapple (<i>Ananas comosus</i>) is a popular fruit worldwide with natural antioxidant properties. This study examined how pineapple modified the expression of drug-metabolizing enzymes (CYP1A2, CYP2C9, CYP3A4, UGT1A6, NAT2 and SULT1A1) and a drug transporter (OATP1B1) in human hepatocarcinoma (HepG2) cells. <b>Materials and Methods:</b> HepG2 cells (2.5×10⁵ cells/well in a 24-well plate) were incubated with pineapple juice extract (125-1,000 µg mL¹) for 48 hrs in phenol red-free medium. Resazurin reduction, ROS, AST and ALT assays were performed. The mRNA expression of target genes was determined by RT/qPCR. <b>Results:</b> Pineapple juice slightly reduced HepG2 cell viability to 80% of the control, while ROS, AST and ALT levels were not changed. Pineapple juice did not alter the expression of CYP1A2, CYP2C9 and UGT1A6 mRNA. All tested concentrations of pineapple juice suppressed CYP3A4, NAT2 and OATP1B1 expression, while SULT1A1 expression was induced. <b>Conclusion:</b> Though pineapple juice slightly decreased the viability of HepG2 cells, cell morphology and cell function remained normal. Pineapple juice disturbed the expression of phase I (CYP3A4) and phase II (NAT2 and SULT1A1) metabolizing genes and the drug transporter OATP1B1. Therefore, the consumption of excessive amounts of pineapple juice poses a risk for drug interactions.

An innovative strategy to rapidly inactivate 8.2-log Enterococcus faecalis in fresh pineapple juice using cold atmospheric plasma.

Enterococcus faecalis is a life-threatening bacterium that resists high levels of antibiotics or chemical preservatives. In this study, we aimed to investigate the inactivation of E. faecalis in fresh pineapple juice (FPJ) with two different cold atmospheric plasmas (CAP) reinforced by H2O2/H2O cold vapor: a plasma jet and a surface dielectric barrier discharge (SDBD). CAP treatments for 300 s with plasma jet and 420 s with SDBD caused an 8.2 log reduction of E. faecalis. The concentration of reactive oxygen and nitrogen species induced in FPJ by plasmas was also evaluated. In terms of quality attributes of FPJ, no noticeable color changes (ΔE) were observed. Furthermore, a trifle of loss of organic content such as sugars and organic acids was observed after treatments. These results suggest that our rapid CAP strategy effectively inactivated E. faecalis in FPJ with no change of color and negligible effects on other physicochemical properties.

Evolution of fungal community associated with ready-to-eat pineapple during storage under different temperature conditions.

The international market of fresh-cut products has witnessed dramatic growth in recent years, stimulated by consumer's demand for healthy, nutritious and convenient foods. One of the main challenging issues for the quality and safety of these products is the potential microbial spoilage that can significantly reduce their shelf-life. The complete identification of fresh-cut product microbiota together with the evaluation of environmental factors impact on microbial composition is of primary importance. We therefore assessed the fungal communities associated with the spoilage of ready-to-eat (RTE) pineapple using a metagenetic amplicon sequencing approach, based on the ITS2 region. Our results revealed a significant variability on fungal species composition between the different batches of RTE pineapple. The initial microbiota composition was the main influencing factor and determined the progress of spoilage. Temperature and storage time were the secondary factors influencing spoilage and their impact was depending on the initial prevalent fungal species, which showed different responses to the various modifications. Our results strongly suggest that further large-scale sampling of RTE pineapple production should be conducted in order to assess the full biodiversity range of fungal community involved in the spoilage process and for unravelling the impact of important environmental factors shaping the initial microbiota.

Gluconacetobacter dulcium sp. nov., a novel Gluconacetobacter species from sugar-rich environments.

A phylogenomic analysis based on 107 single-copy core genes revealed that three strains from sugar-rich environments, i.e. LMG 1728T, LMG 1731 and LMG 22058, represented a single, novel Gluconacetobacter lineage with Gluconacetobacter liquefaciens as nearest validly named neighbour. OrthoANIu and digital DNA-DNA hybridization analyses among these strains and Gluconacetobacter type strains confirmed that the three strains represented a novel Gluconacetobacter species. Biochemical characteristics and MALDI-TOF mass spectra allowed differentiation of this novel species from the type strains of G. liquefaciens and other closely related Gluconacetobacter species. We therefore propose to classify strains LMG 1728T, LMG 1731 and LMG 22058 in the novel species Gluconacetobacter dulcium sp. nov., with LMG 1728T (=CECT 30142T) as the type strain.

Gluconobacter aidae sp. nov., an acetic acid bacterium isolated from tropical fruits in Thailand.

Two bacterial strains, isolates AC10T and AC20, which were reported in a previous study on the diversity of acetic acid bacteria in Thailand, were subjected to a taxonomic study. The phylogenetic analysis based on the 16S rRNA gene sequences showed that the two isolates were located closely to the type strains of Gluconobacter oxydans and Gluconobacter roseus. However, the two isolates formed a separate cluster from the type strains of the two species. The genomic DNA of isolate AC10T was sequenced. The assembled genomes of the isolate were analysed for average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH). The results showed that the highest ANI and dDDH values between isolate AC10T and G. oxydans DSM 3503T were 91.15 and 68.2 %, which are lower than the suggested values for species delineation. The genome-based tree was reconstructed and the phylogenetic lineage based on genome sequences showed that the lineage of isolate AC10T was distinct from G. oxydans DSM 3503T and its related species. The two isolates were distinguished from G. oxydans and their relatives by their phenotypic characteristics and MALDI-TOF profiles. Therefore, the two isolates, AC10T (=BCC 15749T=TBRC 11329T=NBRC 103576T) and AC20 (=BCC 15759=TBRC 11330=NBRC 103579), can be assigned to an independent species within the genus Gluconobacter, and the name Gluconobacter aidae sp. nov. is proposed for the two isolates.

Diversity and Toxigenicity of Fungi that Cause Pineapple Fruitlet Core Rot.

The identity of the fungi responsible for fruitlet core rot (FCR) disease in pineapple has been the subject of investigation for some time. This study describes the diversity and toxigenic potential of fungal species causing FCR in La Reunion, an island in the Indian Ocean. One-hundred-and-fifty fungal isolates were obtained from infected and healthy fruitlets on Reunion Island and exclusively correspond to two genera of fungi: Fusarium and Talaromyces. The genus Fusarium made up 79% of the isolates, including 108 F. ananatum, 10 F. oxysporum, and one F. proliferatum. The genus Talaromyces accounted for 21% of the isolated fungi, which were all Talaromyces stollii. As the isolated fungal strains are potentially mycotoxigenic, identification and quantification of mycotoxins were carried out on naturally or artificially infected diseased fruits and under in vitro cultures of potential toxigenic isolates. Fumonisins B1 and B2 (FB1-FB2) and beauvericin (BEA) were found in infected fruitlets of pineapple and in the culture media of Fusarium species. Regarding the induction of mycotoxin in vitro, F.proliferatum produced 182 mg kg⁻1 of FB1 and F. oxysporum produced 192 mg kg⁻1 of BEA. These results provide a better understanding of the causal agents of FCR and their potential risk to pineapple consumers.

Microbial quality assessment of minimally processed pineapple using GCMS and FTIR in tandem with chemometrics.

Microbial quality is the critical parameter determining the safety of refrigerated perishables. Traditional methods used for assessing microbial quality are time consuming and labour intensive. Thus rapid, non-destructive methods that can accurately predict microbial status is warranted. Models using partial least square regression (PLS-R) from chemical finger prints of minimally processed pineapple during storage obtained by Headspace Solid Phase Microextraction Gas Chromatography Mass Spectrometry (HS-SPME-GCMS), Fourier Transform Infrared (FTIR) spectroscopy and their data fusion are developed. Models built using FTIR data demonstrated good prediction for unknown samples kept under non-isothermal conditions. FTIR based models could predict 87 and 80% samples within ±1 log CFU/g for TVC and Y&M, respectively. Analysis of PLS-R results suggested the production of alcohols and esters with utilization of sugars due to microbial spoilage.

Culture medium for improved production of conidia for identification and systematic studies of Fusarium pathogens.

Fusarium guttiforme and Fusarium ananatum are the etiological agents of fusariosis and fruitlet core rot in pineapple, respectively, producing mycotoxins that are harmful to the health of consumers. These two fungi are morphologically similar and difficulty in obtaining macroconidia of the species limits their identification. Different types of media are available for the culture of these pathogens, but not all of them favor F. ananatum and F. guttiforme macroconidia production. Therefore, the objective of this study was to develop a simple culture medium to improve rapid macro- and microconidia formation in both F. guttiforme and F. ananatum to facilitate taxonomic, pathogenicity and mycotoxin studies. In vitro analysis showed that basal medium with carboxymethyl cellulose (CMC) was better than other media tested with the highest macroconidia production at 7 days of incubation. The highest production of microconidia was with synthetic nutrient medium (SN) at 7 days. F. ananatum produced a relatively high number of microconidia with one septum in comparison to F. guttiforme when cultured in CMC, which suggests an additional character useful for Fusarium taxonomy. CMC medium may serve as an improved alternative to culture media currently used in Fusarium research and contribute to further knowledge of the taxonomy and mycotoxins of Fusarium species.

Mycotoxins from Fusarium proliferatum: new inhibitors of papain-like cysteine proteases.

Papain-like cysteine proteases (PLCPs) in plants are essential to prevent phytopathogen invasion. In order to search for cysteine protease inhibitors and to investigate compounds that could be associated to pineapple Fusarium disease, a chemistry investigation was performed on Fusarium proliferatum isolated from Ananas comosus (pineapple) and cultivated in Czapek medium. From F. proliferatum extracts, nine secondary metabolites were isolated and characterized by nuclear magnetic resonance spectroscopy and mass spectrometry experiments: beauvericin (1), fusaric acid (2), N-ethyl-3-phenylacetamide (3), N-acetyltryptamine (4), cyclo(L-Val-L-Pro) cyclodipeptide (5), cyclo(L-Leu-L-Pro) cyclodipeptide (6), cyclo(L-Leu-L-Pro) diketopiperazine (7), 2,4-dihydroxypyrimidine (8), and 1H-indole-3-carbaldehyde (9). Compounds 1, 3, and 6 showed significant inhibition of papain, with IC50 values of 25.3 ± 1.9, 39.4 ± 2.5, and 7.4 ± 0.5 µM, respectively. Compound 1 also showed significant inhibition against human cathepsins V and B with IC50 of 46.0 ± 3.0 and 6.8 ± 0.7 µM, respectively. The inhibition of papain by mycotoxins (fusaric acid and beauvericin) may indicate a mechanism of Fusarium in the roles of infection process.

Avaliação microbiológica de cookies elaborados com adição do resíduo seco da casca de abacaxi / Evaluation microbiological of cookies prepared with added of residue dry from pineapple peels

A produção de alimentos utilizando subprodutos agroindustriais como a casca de abacaxi enquanto uma das matérias-primas é uma alternativa que proporciona seu aproveitamento, além de contribuir para o valor nutricional do produto. O objetivo do presente estudo foi avaliar a qualidade microbiológica de cookies elaborados com adição do resíduo seco da casca do abacaxi, verificando se o produto obedece aos parâmetros microbiológicos exigidos na legislção vigente. Foram elaborados cookies em 4 formulações distintas, variando a quantidade de resíduo empregada. Através da avaliação microbiológica, quantificou-se os microrganismos do grupo Coliformes, Staphylococcus spp., Salmonella sp., bolores e leveduras. Constatou-se que a manipulação, conservação e higiene durante o preparo dos cookies, foi adequada com relação aos padrões estabelecidos pela legislação.

Caracterização microbiológica de geleias de abacaxi preparadas com açúcar cristal e açúcar demerara / Microbiological characterization of abacaxi jellies prepared with crystal sugar and sugar demerara

O abacaxi é uma das principais frutas cultivadas no Brasil. Diante disso, objetivou-se com esta pesquisa elaborar e avaliar as características microbiológicas de geleias de abacaxi preparadas com açúcar cristal e açúcar demerara. Para a elaboração de cada geleia de abacaxi foi utilizada a proporção de 60 partes da mistura do suco extraído do abacaxi para 40 partes de açúcar e elaborada três formulações, variando a concentração de açúcar cristal e demerara. Após o processamento, foram realizadas as determinações de Coliformes totais e termotolerantes, Salmonella spp., mesófilos, bolores e leveduras. Os resultados indicaram que as geleias elaboradas não apresentaram contaminação por bolores e leveduras, Salmonella spp., coliformes e mesófilos estando em consonância com os parâmetros exigidos pela legislação para geleias.

Characterization and safety evaluation of partially purified bacteriocin produced by Escherichia coli E isolated from fermented pineapple Ananas comosus (L.) Merr.

Antibacterial activity of cell-free supernatant from Escherichia coli E against selected pathogenic bacteria in food and aquaculture was the highest against Edwardsiella tarda 3, a significant aquaculture pathogen. Biochemical properties of the bacteriocins were studied and bacteriocin was found to be sensitive to proteinase K, demonstrating its proteinaceous nature. In addition, pH and temperature affected bacteriocin activity and stability. The bacteriocins were partially purified by ammonium sulfate precipitation. The antibacterial activity was only detected in 20% ammonium sulfate fraction and direct detection of its activity was performed by overlaying on the indicator strains. The inhibition zone associated with the antibacterial activity was detected in the sample overlaid by E. tarda 3 and Staphylococcus aureus DMST8840 with the relative molecular mass of about 27 kDa and 10 kDa, respectively. Bacteriocin showed no cytotoxic effect on NIH-3T3 cell line; however, two virulence genes, aer and sfa, were detected in the genome of E. coli E by PCR. The characteristics of bacteriocins produced by E. coli E exhibited the antibacterial activity against both Gram-positive and Gram-negative pathogenic bacteria and the safe use determined by cytotoxicity test which may have interesting biotechnological applications.

A Rapid and Reliable Method for Molecular Detection of Fusarium guttiforme, the Etiological Agent of Pineapple Fusariosis

Abstract Pineapple (Ananas comosus var. comosus) fusariosis is an economically important fungal disease affecting the plant and its fruit. A rapid and reliable diagnosis is the base of integrated disease management practices. Fusariosis has resulted in quarantines for pineapple products in Central America, Africa and Asia. Difficulties diagnosing and correctly identifying the fungus Fusarium guttiforme, agent of the pineapple fusariosis, have led to the search for new methodologies, and for this we developed a new reliable molecular method to detect it. For diagnostic purposes, real-time PCR of elongation factor gene 1-α (ef1) was used to rapidly, specifically and sensitively diagnose F. guttiforme. A pathogenicity test was conducted with slips of the pineapple cultivar Pérola, a multiplex PCR was run, and the results compared with those obtained with real-time PCR. The real-time PCR assay with its specific primer set could readity distinguish F. guttiforme from other Fusarium species known to occur on pineapple. The real-time PCR test had 95% sensitivity and 100% specificity with a significance level p<0.0001. For field samples the test had 100% sensitivity and specificity. Thus, this new test is fit for use in serial analyses of pineapples, and may have application in the evaluation of propagation materials and making quarantine decisions. The ability to rapidly and specifically detect F. guttiforme in plant samples will facilitate monitoring of the pathogen and improve disease management.

Investigation of damage to Escherichia coli, Listeria monocytogenes and Salmonella Enteritidis exposed to Mentha arvensis L. and M. piperita L. essential oils in pineapple and mango juice by flow cytometry.

The effects of Mentha arvensis L. (MAEO; 0.625 µL/mL) and M. piperita L. (MPEO; 1.25 µL/mL) essential oils on viable cell counts and physiological functions in Escherichia coli, Listeria monocytogenes and Salmonella enterica Serovar Enteritidis in pineapple and mango juice after a 15 min-exposure under refrigeration were evaluated in this study. The physiological functions of the bacterial cells were assessed by flow cytometry using the fluorochromes thiazole orange, propidium iodide, bis-1,3-dibutylbarbutiric acid, ethidium bromide, and 5-cyano-2,3-ditolyl tetrazolium chloride to investigate membrane integrity, membrane potential, efflux activity, and respiratory activity. MAEO and MPEO sharply reduced (>5 log10 CFU/mL cycles) the counts of E. coli, L. monocytogenes and Salmonella Enteritidis in pineapple juice, and caused smaller reductions (0.61-1.58 log10 CFU/mL cycles) in mango juice. Bacterial cells exposed to MAEO and MPEO in pineapple and mango juice showed increased membrane permeability, membrane depolarization and changes in efflux pump and respiratory activity. More physiological damage occurred in bacterial cell populations exposed to MAEO or MPEO in pineapple juice than in mango juice. These results indicate that MAEO and MPEO inactivate E. coli, L. monocytogenes and Salmonella Enteritidis cells in pineapple and mango juice through a multi-target action mode that disrupts cytoplasmic membranes, increases permeability and potential depolarization, as well as inhibits efflux pump and respiratory activity.

Inactivation of the microbiota and effect on the quality attributes of pineapple juice using a continuous flow ultrasound-assisted supercritical carbon dioxide system.

Supercritical carbon dioxide inactivation technology represents a promising nonthermal processing method, as it causes minimum impact on the nutritional food properties. The aim of this study was to analyze the combined effect of supercritical carbon dioxide and high-power ultrasound on the inactivation of natural microbiota and the quality attributes of pineapple juice treated in a continuous flow system. Different juice residence times (3.06-4.6 min), at 100 bar and 31.5 ℃, were used. The results indicated that the microbiota inactivation was complete and the differences obtained in the quality attributes (2.2% for pH, 4.8% for °Brix, 2% for vitamin C) were minimal. During storage, microorganisms were not able to recover and the vitamin C decrease could be limited to 8.2% after four weeks. The results demonstrated that the supercritical carbon dioxide-high-power ultrasound technique could be an excellent alternative for the cold pasteurization of pineapple juice.

Characterization of highly branched dextran produced by Leuconostoc citreum B-2 from pineapple fermented product.

A strain Leuconostoc citreum B-2 was isolated from homemade fermentation product of pineapple and its polysaccharide yield was 28.3g/L after cultivating the strain in Man-Rogosa-Sharpe (MRS) medium with 75g/L sucrose. The exopolysaccharide (EPS) was characterized by gas chromatography (GC), Fourier-transform infrared (FT-IR) spectra, high-performance size-exclusion chromatography (HPSEC), nuclear magnetic resonance (NMR) spectroscopy and scanning electron microscope (SEM) analysis in present study. The monosaccharide composition of EPS was glucose and molecular weight was 3.77×106Da. FT-IR and NMR spectra revealed that the B-2 EPS was composed of 75% α-(1→6) linked d-glucopyranose units existing in the main chain with 19% α-(1→3) branching and only a few α-(1→2) branching. The SEM of the dried EPS appeared irregular sheets with glittering surface and compact structure. Water solubility index and water holding capacity of B-2 EPS were 80% and 450%, respectively. All the mentioned characteristics suggested that the EPS has a potential application in the food, cosmetic and pharmaceuticals industry.

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.

Evaluating the response to Fusarium ananatum inoculation and antifungal activity of phenolic acids in pineapple.

Fusarium ananatum causes fruitlet core rot (FCR) in pineapple (Ananas comosus var. comosus) when the fruit reaches maturity. Hidden symptoms make it difficult to assess the disease, regardless of its stage, and basic questions concerning the involvement of the phenolic compounds in response to infection remain unknown. A direct inoculation method of F. ananatum in pineapple fruitlets was developed to monitor the growth of black spots and the changes in phenolic acids and ascorbic acid concentration under controlled conditions. After inoculation, infection began with a flesh discolouration at the inoculation point and then spread in a darker shade to form a black spot. Coumaroyl-isocitric and caffeoyl-isocitric acids levels respectively showed a 150- and 200-fold increase in infected fruitlet when compared to healthy fruitlet. These hydroxycinnamic acids increased minimally in the adjacent fruitlet and remained stable in the other parts of the fruit. By contrast, sinapic acid and hydroxybenzoic acid isomers (HBA) decreased after F. ananatum inoculation in the infected fruitlet, whereas they remained stable in the adjacent and healthy fruitlets. Ascorbic acid decreased to zero in the infected fruitlet. The antifungal activity of phenolic compounds and ascorbic acid was evaluated against the mycelial growth of F. ananatum. p-Coumaric acid exhibited a total inhibition of the mycelial growth at 1000 µg g-1. Ferulic acid inhibited 64 % of mycelial growth at a concentration of 1000 µg g-1. Caffeoylquinic acid, sinapic acid, and ascorbic acid also showed significant antifungal activity, but to a lesser extent. Finally, coinoculation of the hydroxycinnamic acids with the pathogen restrains its development in the fruit. This is the first study to highlight the involvement of phenolic compounds in the pineapple FCR disease.

Análise microbiológia da casca do abacaxi (Ananás comosus) sob diferentes condições de higienização para obtenção de suco e chá / Microbiological analysis of pineapple bark (Ananás comosus) under different hygienic conditions to obtain juice and tea

A casca do abacaxi é um resíduo da agroindústria que pode auxiliar na dieta humana de forma complementar, permitindo sua utilização em alimentos de baixo valor nutricional. Percebendo que a casca do abacaxi é desprezada pelas indústrias de alimentos e devido à grande preocupação global atual de se minimizar os resíduos buscando uma melhoria do meio ambiente e de geração de lucros para empresas, neste estudo teve-se por objetivo avaliar os micro-organismos existentes na casca do abacaxi, antes e após a higienização e sanitização para o preparo de sucos e chás. Na higienização foi utilizada água corrente e na sanitização solução clorada a 200 ppm. Com os dados obtidos concluiu-se que os cuidados com a higienização adequada da casca são importantes, que a lavagem somente com água não é eficaz para a eliminação total dos micro-organismos presentes na casca do abacaxi, já a higienização com solução clorada é amplamente recomendada para retardar ou eliminar o crescimento microbiano.