RESUMO
The paper presents data on phytosanitary monitoring of garden cenoses for fire blight in the Turkestan, Zhambyl, and Almaty regions of Kazakhstan. The purpose of this study is to assess the phytosanitary situation in various regions of Kazakhstan, determine the extent of fire blight spread, and isolate and identify the fire blight pathogen. During the study, methods such as hypersensitivity, pathogenicity, and fluorescent simplification-based specific hybridization polymerase chain reaction (FLASH-PCR) were used. It was found that in all the surveyed areas, disease foci were identified. For the first time, the fire blight pathogen was detected on fruit crops such as plum, peach, cherry plum, and quince, as well as on wild apricots. 274 plant samples were collected from which microorganisms were isolated. Isolates related to the fire blight pathogen Erwinia amylovora were identified by methods of hypersensitivity, pathogenicity, and FLASH-PCR diagnostics. Of the 156 isolates of microorganisms isolated from apple tree plant samples, 21 inhibited the in vitro growth of E. amylovora to varying degrees. Isolates 16.2 and 19.2 with maximum antagonistic activity were selected, where the pathogen growth inhibition zones were 52.2 ± 2.58 mm and 45.6 ± 0.55 mm, respectively. Based on the obtained sequence of nucleotides of the 16SpRNA gene site, it was found that the selected isolates with high antagonistic activity belonged to the Pseudomonas genus. In the future, based on these isolates, a new biological product for fire blight control can be created and adapted to the natural and climatic conditions of Kazakhstan.
Assuntos
Erwinia amylovora , Doenças das Plantas , Cazaquistão , Doenças das Plantas/microbiologia , Erwinia amylovora/isolamento & purificação , Reação em Cadeia da Polimerase , Malus/microbiologiaRESUMO
Alicyclobacillus spp. is the cause of great concern for the food industry due to their spores' resistance (thermal and chemical) and the spoilage potential of some species. Despite this, not all Alicyclobacillus strains can spoil fruit juices. Thus, this study aimed to identify Alicyclobacillus spp. strains isolated from fruit-based products produced in Argentina, Brazil, and Italy by DNA sequencing. All Alicyclobacillus isolates were tested for guaiacol production by the peroxidase method. Positive strains for guaiacol production were individually inoculated at concentration of 103 CFU/mL in 10 mL of orange (pH 3.90) and apple (pH 3.50) juices adjusted to 11°Brix, following incubation at 45 °C for at least 5 days to induce the production of the following spoilage compounds: Guaiacol, 2,6-dichlorophenol (2,6-DCP) and 2,6-dibromophenol (2,6-DBP). The techniques of micro-solid phase extraction by headspace (HS-SPME) and gas-chromatography with mass spectrometry (GC-MS) were used to identify and quantify the spoilage compounds. All GC-MS data was analyzed by principal component analysis (PCA). The effects of different thermal shock conditions on the recovery of Alicyclobacillus spores inoculated in orange and apple juice (11°Brix) were also tested. A total of 484 strains were isolated from 48 brands, and the species A. acidocaldarius and A. acidoterrestris were the most found among all samples analyzed. In some samples from Argentina, the species A. vulcanalis and A. mali were also identified. The incidence of these two main species of Alicyclobacillus in this study was mainly in products from pear (n = 108; 22.3 %), peach (n = 99; 20.5 %), apple (n = 86; 17.8 %), and tomato (n = 63; 13 %). The results indicated that from the total isolates from Argentina (n = 414), Brazil (n = 54) and Italy (n = 16) were able to produce guaiacol: 107 (25.8 %), 33 (61.1 %) and 13 (81.2 %) isolates from each country, respectively. The PCA score plot indicated that the Argentina and Brazil isolates correlate with higher production of guaiacol and 2,6-DCP/2,6-DBP, respectively. Heatmaps of cell survival after heat shock demonstrated that strains with different levels of guaiacol production present different resistances according to spoilage ability. None of the Alicyclobacillus isolates survived heat shocks at 120 °C for 3 min. This work provides insights into the incidence, spoilage potential, and thermal shock resistance of Alicyclobacillus strains isolated from fruit-based products.
Assuntos
Alicyclobacillus , Sucos de Frutas e Vegetais , Frutas , Cromatografia Gasosa-Espectrometria de Massas , Guaiacol , Esporos Bacterianos , Alicyclobacillus/isolamento & purificação , Alicyclobacillus/genética , Alicyclobacillus/classificação , Alicyclobacillus/crescimento & desenvolvimento , Sucos de Frutas e Vegetais/microbiologia , Guaiacol/análogos & derivados , Guaiacol/metabolismo , Guaiacol/farmacologia , Frutas/microbiologia , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Bacterianos/isolamento & purificação , Microbiologia de Alimentos , Contaminação de Alimentos/análise , Brasil , Microextração em Fase Sólida , Argentina , Malus/microbiologia , Itália , Temperatura Alta , Citrus sinensis/microbiologiaRESUMO
Apples (Malus domestica) are one of the most consumed fruits globally. It is a relevant crop in Argentina and Spain, and one of the main fruits for export and industrialization in these countries. Quality control of apples, fundamentally in the postharvest stage, is critical to prevent fungal diseases. The blue mould, caused by Penicillium expansum, is responsible for great economic losses due to the deterioration of the fruit and mycotoxin production. Many studies have characterized this pathogen; however, little is known about the differences between populations from distant geographical origins. The objective of the present study was to characterize two P. expansum populations, from Argentina and Spain, through morphological, metabolomic and molecular approaches, and to evaluate the existence of differences related to their geographical source. A total of 103 isolates, 53 from Argentina and 50 from Spain were studied. Their morphological features were consistent with the species description. The secondary metabolite profiles revealed low chemical diversity. All 103 isolates shared the production of 13 compounds, namely andrastins, aurantioclavine, chaetoglobosins, communesins, expansolides, roquefortine C and patulin. Penostatins and citrinin were produced by 102 and 101 isolates, respectively. A region of the ß-tubulin gene was selected to analyse the diversity of the P. expansum isolates. No substantial differences were observed between isolates of different geographical origins through morphology, patulin accumulation, secondary metabolite profiles and phylogenetic analysis. However, the analysis of polymorphisms revealed 29 haplotypes with a relative separation between isolates of both populations; 13 haplotypes contained Argentinean isolates, while Spanish isolates were separated into 16 haplotypes. The diversity indices of Shannon (H'=2.075; H'=2.402) and Simpson (SiD = 0.850; SiD = 0.895) for isolates from Argentina and Spain, respectively, indicated that the diversity of P. expansum is greater in Spain than in Argentina. This distribution could be explained both by the existence of haplotype exchange between both countries, with the ancestral haplotypes originating in Spain, and the subsequent adaptation to the environmental conditions or apples varieties grown in each region.
Assuntos
Malus , Patulina , Penicillium , Argentina , Frutas/microbiologia , Malus/microbiologia , Patulina/análise , Penicillium/genética , Penicillium/metabolismo , Filogenia , EspanhaRESUMO
Alternaria is a frequent contaminant of apple fruit, causing severe economic losses. It can produce external lesions and mouldy core, characterised by a rotten area in the apple core. In the present study, morphological and chemical characterization of Alternaria from apples was performed, evaluating differences related to agricultural practices and type of disease. A low morphological diversity was observed; most of the isolates were identified as A. tenuissima sp.-grp. (95 %). A. arborescens sp.-grp. and A. gaisen sp.-grp. were present in a proportion of 1 %, and 3 % of the isolates showed intermediate characteristics between these sp.-grps. and were identified as Alternaria sp. The chemical diversity was greater; 27 secondary metabolites were produced by the apple isolates. The most frequents were altertoxin-I (85 %), altechromone A (76 %), tentoxin (69 %), and tenuazonic acid (68 %). The alternariols were produced in a lower frequency when comparing with isolates from other crops; alternariol, 58 % and alternariol monomethyl ether, 57 %. The predominant secondary metabolite profile included compounds from different chemical families, such as dibenzopyrones, tetramic acids, perylene quinones, and cyclic tetrapeptides. A wider metabolomic capacity was observed in isolates from conventional apples when compared to those from organic fruit, with the predominance of strong producers of altertoxins and alternariols. The isolates from mouldy core showed higher ability to produce metabolites from different chemical families than those from external lesions. The wide chemical diversity of the Alternaria apple population should be considered to assess the health risk associated with apple by-products.
Assuntos
Alternaria , Malus , Micotoxinas , Alternaria/metabolismo , Produtos Agrícolas , Frutas/química , Malus/microbiologia , Micotoxinas/análise , Doenças das Plantas , Ácido Tenuazônico/análiseRESUMO
Bacteria were isolated from samples of Fresh Apple juices from shops of three different localities of Lahore. Analysis of samples from Liberty, Anarkali and Yateem khana Markets show different levels of contamination. There were pathogenic and non-pathogenic bacteria in all samples and were identified by the morphological and biochemical tests. Most of the plasmids of pathogenic bacteria were 4kb in their molecular size. Ribotyping of 16S ribosomal RNA gene sequencing was done to confirm Helicobacter pylori strain and Gluconobacter oxydans. The highest sensitivity of 210mm was shown by Enterobacter sp. against Aztheromysine disk (15µg) while Micrococcus sp. was highly resistant against all of the Antibiotics applied. The antibiotic resistance of pathogenic bacteria was also checked against Ricinus communis plant's extracts, all isolated bacterial pathogens were resistant but only, E.coli was inhibited at 300µl of the extracts. Presence of pathogenic bacteria in Apple juice samples was due to contamination of sewage water in drinking water while some of these pathogenic bacteria came from Apple's tree and other from store houses of fruits.
Assuntos
Antibacterianos , Gluconobacter oxydans , Helicobacter pylori , Extratos Vegetais , Ricinus/química , Antibacterianos/farmacologia , Sucos de Frutas e Vegetais/microbiologia , Gluconobacter oxydans/efeitos dos fármacos , Helicobacter pylori/efeitos dos fármacos , Malus/microbiologia , Testes de Sensibilidade Microbiana , Extratos Vegetais/farmacologiaRESUMO
In the present work, we evaluated the effects of a mixture of biocontrol agents against two toxigenic strains of Penicillium expansum isolated in Argentine Patagonia from pome fruits. The two strains, INTA-5 and INTA-10, were previusly selected among ten strains coming from the Alto Valle (Rio Negro-Argentina) for their high production of patulin. For the biocontrol, Kosakonia radicincitans, Cryptococcus laurentii, and Rhodosporidium fluviale were tested in vitro experiments on Potato Dextrose Agar (PDA) dishes against the INTA-5 and INTA-10 strains. The bacterium K. radicincitans and the yeast C. laurentii were selected to be used in a mixture due to their capacity to control the fungus and reduce the mycotoxin severely. In vitro assays with the mixture showed a high antagonism against P. expansum INTA-5 and INTA-10, at 21 d of incubation at 25 °C and a patulin reduction of 98%. The mixture of microorganisms was also effective in apples stored at 25 °C for 10 d and 4 °C for 30 d. At cold storage, the mixture controlled moderately the development of rot and decreased patulin concentration. At 25 °C, the pathogen's optimal growth temperature, the mixture of Biological Control Agent (BCAs) assured both the control of rot and decrease of patulin concentration. The combination of two microorganisms, with different requirements and abilities, resulted in a mix with a strong antagonism against P. expansum with the capability to decrease the patulin concentration. Treatment with the selected mixture could be a good option for controlling strains with different behaviours and in different environmental conditions.
Assuntos
Antibiose , Agentes de Controle Biológico/farmacologia , Cryptococcus/fisiologia , Enterobacteriaceae/fisiologia , Malus/microbiologia , Patulina/biossíntese , Penicillium/efeitos dos fármacos , Penicillium/metabolismo , Doenças das Plantas/microbiologia , Frutas/microbiologiaRESUMO
The isolation of autochthonous yeast species presents a good strategy to select new microorganisms for developing an adequate inoculum to carry out fermentations and generate representative products of the cider production zone. However, non-Saccharomyces yeasts have been considered to have low capacity to carry out a complete fermentation as Saccharomyces cerevisiae. In this work, five autochthonous yeasts from a cider fermentation process were isolated and identified as Saccharomyces cerevisiae, Kluyveromyces marxianus, Pichia membranaefaciens, P. kluyveri and Zygosaccharomyces rouxii. A series of fermentations were developed at laboratory level, using each species individually and it was observed that only S. cerevisiae was able to finish the process. K. marxianus consumed less than 50% of the sugars; P. kluyveri and Z. rouxii consumed less than 70% and P. membranaefaciens consumed more than 90% but the yield (ethanol produced for sugar consumed (YP/S)) was 0.39. Nevertheless, the addition of magnesium, zinc and nitrogen increased the fermentative capacity of almost all species: K. marxianus, Z. rouxii and P. kluyveri, showed an increase in ethanol production when nutrients were added, obtaining more than 80 g/L of ethanol, and showing that those nutrients are necessary to complete the fermentation. This work describes the potential use of different non-Saccharomyces species to carry out fermentation of apple juice and highlights the importance of certain nutrients to enable an efficient alcoholic fermentation and the generation of desirable volatile compounds for cider production.
Assuntos
Bebidas Alcoólicas/microbiologia , Nutrientes/metabolismo , Leveduras/metabolismo , Bebidas Alcoólicas/análise , Etanol/análise , Etanol/metabolismo , Fermentação , Sucos de Frutas e Vegetais/análise , Sucos de Frutas e Vegetais/microbiologia , Malus/microbiologia , Nutrientes/análise , Especificidade da Espécie , Leveduras/classificação , Leveduras/isolamento & purificaçãoRESUMO
The production of pome fruits as pears and apples, as well as their derived industries, is of great economic importance in North Patagonia. The elaboration of fermented beverages as cider or perry has evidenced a substantial diversification during the last years, with the evaluation of different fruit varieties, yeast starters and technological changes. In this work, two cryotolerant yeasts belonging to the species Saccharomyces uvarum were evaluated at laboratory and pilot scale in sterile and no-sterile pear must. One of the strains was originally isolated from apple chicha (strain NPCC1314) and the other from apple cider (strain NPCC1420) in Patagonia. Both physicochemical and sensory features of the fermented products were evaluated. Both strains were able to successfully complete the fermentations, although strain NPCC1420 showed the better kinetic properties including a faster sugar consumption than the strain NPCC1314. Both strains showed excellent implantation capacity, but the fermented products showed different chemical profiles. The perry fermented with the strain NPCC1314 was characterized by better sensory attributes as assessed by trained panelists and a greater acceptance for untrained public than the same fermented with the strain NPCC1420. The two strains were able to consume sorbitol, both in pear must and in agar-plates supplemented with sorbitol as the sole carbon source. This ability is described for the first time in S. uvarum, at least for the two strains evaluated in this work.
Assuntos
Bebidas Alcoólicas/análise , Alimentos Fermentados/microbiologia , Pyrus/microbiologia , Saccharomyces/metabolismo , Argentina , Reatores Biológicos , Chile , Fermentação , Frutas/química , Frutas/microbiologia , Malus/microbiologia , Saccharomyces/isolamento & purificação , Leveduras/classificação , Leveduras/isolamento & purificaçãoRESUMO
BACKGROUND: In recent years the use of high-pressure processing (HPP) of fruit products has steadily increased due to its antimicrobial effectiveness and the retention of nutritional and quality attributes compared to conventional thermal technologies. Edible coatings are already being used to enhance the quality of minimally processed fruits. Thus, apple cubes (AC) and alginate-vanillin-coated apple cubes (AVAC) were subjected to HPP (400 MPa/5 min/35 °C). The microbiological and physicochemical parameters were evaluated and the bioactive compounds were monitored before and after HPP of apple cubes. Also, an in vitro gastrointestinal digestion (GID) was conducted. RESULTS: HPP left L. monocytogenes counts below the detection limit (2 log UFC g-1 ), regardless of the presence of coating. For E. coli, HPP + active coating showed a synergism affording the greatest reduction (>5 log) for AVAC-HPP. Firmness was maintained in AVAC-HPP samples, while AC-HPP samples suffered reductions of 35%. Colour attributes were also better retained in AVAC-HPP samples. In general, HPP led to a decrease in phenolic compounds. Regarding the effects of GID, vanillin-based active coating exerted a protective effect on some phenolics. Thus, p-coumaroylquinic acid concentration was maintained for AVAC and AVAC-HPP during GID. Epigallocatechin, the compound with the highest concentration in apple cubes, increased for AVAC (106%) and AVAC-HPP (57%). Also, phloridzin concentration increased for AVAC-HPP (17%). At the end of GID, procyanidin B1 and epigallocatechin were the main phenolic compounds for all samples, AVAC showing the highest concentration. CONCLUSIONS: This work demonstrates that the combined application of HPP and active coatings on apple cubes could be used to obtain a safe and good-quality product. © 2021 Society of Chemical Industry.
Assuntos
Conservação de Alimentos/métodos , Conservantes de Alimentos/farmacologia , Frutas/microbiologia , Malus/química , Fenóis/química , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Conservação de Alimentos/instrumentação , Frutas/química , Malus/microbiologia , Valor NutritivoRESUMO
The estimation of growth or inactivation of bacterial population in fruits during preservation and storage provides useful information for the improvement of the safety of fresh-cut fruits and vegetables. This paper addressed the attachment to the surface and the growth in the flesh of apple fruits of four bacterial cultures (Escherichia coli, Bacillus cereus, Staphylococcus aureus and Pseudomonas aeruginosa). The growth of the bacterial cultures in apple flesh was monitored at particular time intervals, and Gompertz parameters, i.e. maximum number of bacteria (Pm), the maximum growth rate of bacteria rp,m, and lag time tl, were used to determine the growth kinetics. After the immersion, the highest number of P. aeruginosa and the lowest number of B. cereus adhered to the apples. After washing and swabbing, E. coli was reduced from the surface of apples to the highest extent (by 3.34 log cfu g-1), while the number of B. cereus was reduced to the lowest extent (1.66 log cfu g-1). Fitted curves of the Gompertz model corresponded quite well to the measured values of the number of microorganisms with R2 = 0.92-0.98. The values of the standard error (0.17-0.37) and extremely low p values of the Fischer test (p < 0.0001) indicated strict dependence between the model predicted and the maximum population density. The predicted values of the maximum number of microorganisms (Pm) correspond almost exactly to the actual values. A similar conclusion can be drawn for the maximum growth rate of microorganisms (rp,m), with the measured value being slightly higher than predicted values.