Evaluation of process involved in the production of aromatic compounds in Gram-negative bacteria isolated from vanilla (Vanilla planifolia ex. Andrews) beans.

AIM: The present investigation was aimed at isolating and identifying bacterial strains from cured vanilla beans. Additionally, the study focused on evaluating bacterial processes pertaining to the aromatic compounds production (ACP). METHODS AND RESULTS: Three bacteria were isolated from Vanilla planifolia beans, previously subjected to the curing process. According to morphological, biochemical and 16S rRNA analysis, the strains were identified as Citrobacter sp., Enterobacter sp. and Pseudomonas sp. The polygalacturonase activity (PGA) was determined using the drop, cup-plate and DNS methods. Aromatic compounds production was analysed by cup-plate method using FA as substrate and quantified by high performance liquid chromatography (ppm), the functional groups of vanillic acid (VA) were identified by FT-IR and the aromatic compounds (AC) resistance was determined and reported as minimum inhibitory concentration. Citrobacter sp., Enterobacter sp. and Pseudomonas showed PGA (70·31 ± 364, 76·07 ± 12·47 and 51 ± 10·92 U ml-1 respectively), were producers of VA (3·23 ± 0·49, 324 ± 41 and 265·99 ± 11·61 ppm respectively) and were resistant to AC. CONCLUSIONS: The Gram-negative bacteria isolated from V. planifolia beans were responsible for ACP. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first evidence for the role of Gram-negative bacterial isolates from cured Mexican V. planifolia beans in the process related to ACP.

Thermosonication parameter effects on physicochemical changes, microbial and enzymatic inactivation of fruit smoothie.

With the aim of developing a fruit-based beverage in products which are severely damaged by heat, a high-intensity ultrasound treatment combined with moderate heat treatment (called thermosonication) was applied. A fruit smoothie (mango, jackfruit and rice milk) was thermosonicated applying a Box-Benhken model with amplitude (70, 77.5 or 85%), time (15, 20 or 25 min) and temperature (40, 47.5 or 55 °C) as independent variables. From the obtained samples, microbiological (aerobic mesophilic and Enterobacteriaceae), physicochemical (pH, soluble solids and cloud index) and enzymatic analysis (polyphenol oxidase and pectin methylesterase) were carried out. Aerobic mesophiles and Enterobacteria inactivation in thermosonicated samples were 4.55 Log CFU/mL and 3.85 Log CFU/mL, respectively in most of the treatments applied, being influenced by linear terms of amplitude and temperature (p < 0.001). The cloud index was influenced by time term (p < 0.0001); meanwhile, interaction of amplitude * temperature (p < 0.01) and quadratic of time presented significant effect (p < 0.001) on polyphenol oxidase activity. Further, amplitude term had a significant effect (p < 0.001) on the decrease on pectin methylesterase enzymatic activity. The optimal process condition was 77.5% amplitude, 20 min and 47.5 °C. Thermosonication probed to be effective to control both enzymatic activities in treatments with high amplitudes combined with moderated temperature treatments. Based on this, the use of thermosonication is a viable alternative for fruit-based beverage preservation, that may employ perishable regional natural products offering them an added value.

High pressure processing effect on different Listeria spp. in a commercial starter-free fresh cheese.

In this study, both microbial inactivation and growth of Listeria spp. inoculated in commercial free-starter fresh cheese was evaluated after high-pressure processing (HPP). HPP conditions (300, 400, 500 and 600 MPa at 6 °C for 5 min) and inoculum level (3-4 or 6-7 log CFU/g of cheese), as well as differences among strains inoculated (Listeria innocua, L. monocytogenes CECT 4031 and L. monocytogenes Scott A) were investigated. Inactivation and generation of sublethal injury were determined after HPP using ALOA (Agar Listeria according to Ottaviani and Agosti) and TAL (Thin Agar Layer) plating methods, respectively. Listeria inactivation increased with the pressure applied, presenting some statistical differences between the employed strains, inoculum level and sublethal injury. The highest lethality values were obtained at 600 MPa for the three strains tested, although the 500 MPa treatment presented high lethality for L. innocua and L. monocytogenes CECT 4031. After treatment, L. innocua and L. monocytogenes CECT 4031 counts in fresh cheese increased gradually during cold storage. By contrast, counts in cheeses inoculated with L. monocytogenes Scott A did not change significantly (p ≥ 0.05), being this strain the most pressure resistant and with the slowest growth rate. The manuscript present information supporting that, strains with high-level resistance should be employed during inactivation studies, instead of surrogate microorganisms. Application of HPP treatments of 500 MPa and especially 600 MPa on fresh cheeses would be effective to eliminate the most resistant microorganism to a level that should not present a public health risk under normal conditions of distribution and storage.

Effect of moderate inlet temperatures in ultra-high-pressure homogenization treatments on physicochemical and sensory characteristics of milk.

The effect of ultra-high-pressure homogenization (UHPH) on raw whole milk (3.5% fat) was evaluated to obtain processing conditions for the sterilization of milk. Ultra-high-pressure homogenization treatments of 200 and 300 MPa at inlet temperatures (Ti) of 55, 65, 75, and 85 °C were compared with a UHT treatment (138 °C for 4s) in terms of microbial inactivation, particle size and microstructure, viscosity, color, buffering capacity, ethanol stability, propensity to proteolysis, and sensory evaluation. The UHPH-treated milks presented a high level of microbial reduction, under the detection limit, for treatments at 300 MPa with Ti of 55, 65, 75, and 85 °C, and at 200 MPa with Ti = 85 °C, and few survivors in milks treated at 200 MPa with Ti of 55, 65, and 75 °C. Furthermore, UHPH treatments performed at 300 MPa with Ti = 75 and 85 °C produced sterile milk after sample incubation (30 and 45 °C), obtaining similar or better characteristics than UHT milk in color, particle size, viscosity, buffer capacity, ethanol stability, propensity to protein hydrolysis, and lower scores in sensory evaluation for cooked flavor.

Multiple-pass high-pressure homogenization of milk for the development of pasteurization-like processing conditions.

Multiple-pass ultrahigh pressure homogenization (UHPH) was used for reducing microbial population of both indigenous spoilage microflora in whole raw milk and a baroresistant pathogen (Staphylococcus aureus) inoculated in whole sterile milk to define pasteurization-like processing conditions. Response surface methodology was followed and multiple response optimization of UHPH operating pressure (OP) (100, 175, 250 MPa) and number of passes (N) (1-5) was conducted through overlaid contour plot analysis. Increasing OP and N had a significant effect (P < 0·05) on microbial reduction of both spoilage microflora and Staph. aureus in milk. Optimized UHPH processes (five 202-MPa passes; four 232-MPa passes) defined a region where a 5-log(10) reduction of total bacterial count of milk and a baroresistant pathogen are attainable, as a requisite parameter for establishing an alternative method of pasteurization. Multiple-pass UHPH optimized conditions might help in producing safe milk without the detrimental effects associated with thermal pasteurization.

High-power ultrasound bleaching technique for canola oil (Brassica napus L.): Pigments removal and quality parameters.

Canola seeds (Brassica napus L.) are among the most commonly used seeds in Mexico for vegetable oil production. This is based on the high yield and content of polyunsaturated and monounsaturated fatty acids. During oil bleaching, it is important to maintain fatty acids in their cis configuration because of the health concerns associated with trans-fatty acid consumption. In this sense, the industrial processing parameters employed for this purpose present some limitations, such as high temperatures and long times, which may change the cis configuration to trans. In addition, the amount of bleaching clay employed for this process could be a source of contamination because it is disposed of after treatment. Therefore, the aim of this study was to develop a bleaching process for canola oil using high-power ultrasound (US). US processing was applied to nine treatments with different processing times (60, 75, and 90 min), clay percentages (1, 2, and 3%), and temperatures (60 and 80 °C) to determine the concentrations of chlorophyll a and b (µg chlo/100 g oil), carotenes (µg ß-carotene/100 g oil), color (L*, a*, b*, C*, and h°), iodine value (g I2/100 g oil), and finally carrying out a spectroscopic analysis (ATR-FTIR and Raman). A conventional bleaching treatment (100 °C for 180 min, 3% bleaching clay) was used as a control. The results revealed that US treatments with 2% clay at 60 °C for 60 and 90 min eliminated most of the chlorophyll compounds (98%). However, in terms of carotenes reduction, these identical treatments exhibited a similar tendency to that of the control (approximately 30% decrease). These findings also affected the sample color, in which US treatments revealed chromatic coordinates that indicated yellow tones with chroma values that were more intense than those in the control samples. In terms of the iodine value, such treatments fulfilled the international standards for vegetable oils (90-100 g I2/100 g oil). Finally, the spectroscopic study revealed no trans configurations or the presence of different chemical compounds after US treatment, because neither of them presented typical peaks for those molecular configurations. In this regard, US can be a useful methodology for bleaching vegetable oils, helping to reduce time, and bleaching clay with similar pigment reduction results.

High-intensity ultrasound pretreatment influence on whey protein isolate and its use on complex coacervation with kappa carrageenan: Evaluation of selected functional properties.

The aim of this work was to evaluate the influence of high-intensity ultrasound (HIUS) treatment on whey protein isolate (WPI) molecular structure as a previous step for complex coacervation (CC) with kappa-carrageenan (KC) and its influence on CC functional properties. Protein suspension of WPI (1% w/w) was treated with an ultrasound probe (24 kHz, 2 and 4 min, at 50 and 100% amplitude), non HIUS pretreated WPI was used as a control. Coacervation was achieved by mixing WPI and KC dispersions (10 min). Time and amplitude of the sonication treatment had a direct effect on the molecular structure of the protein, FTIR-ATR analysis detected changes on pretreated WPI secondary structure (1600-1700 cm-1) after sonication. CC electrostatic interactions were detected between WPI positive regions, KC sulfate group (1200-1260 cm-1), and the anhydrous oxygen of the 3,6 anhydro-D-galactose (940-1066 cm-1) with a partial negative charge. After ultrasound treatment, a progressive decrease in WPI particle size (nm) was detected. Rheology results showed pseudoplastic behavior for both, KC and CC, with a significant change on the viscosity level. Further, volume increment, stability, and expansion percentages of CC foams were improved using WPI sonicated. Besides, HIUS treatment had a positive effect on the emulsifying properties of the CC, increasing the time emulsion stability percentage. HIUS proved to be an efficient tool to improve functional properties in WPI-KC CC.