Plate Count Research Articles

Microchip gas chromatographic columns dedicated for space exploration: Stationary phase coating, setup optimization and evaluation of column performances

This work is the first part of a project aiming to specifically design gas chromatographic microcolumns for space exploration. In particular, this study explored the functionalization and characterization of silicon/glass microchip gas chromatographic columns designed with a serpentine-shaped channel having an internal square cross-section. This microcolumn can be connected using a robust fluidic manifold with removable capillary connections to a conventional gas chromatograph or implemented in a prototype instrument for space exploration. First, benchtop gas chromatographic system was optimized in terms of injector liner volume, internal diameter (I.D.) of the capillary connections and data frequency of detection to ensure an optimal evaluation of column efficiency. Junction capillaries of 100 μm I.D., a liner of 1.2 mm I.D. and a detection acquisition frequency of 100 Hz were found to be the optimal set-up and parameters. Moreover, the stationary phase coating velocity was slowed-down to increase column efficiency by 100 %. Then, the performances of a square cross-section capillary column were compared with those of a conventional circular cross-section column. The coating efficiencies were estimated at 55 % and 42 % for circular and square internal cross-section geometries respectively, demonstrating that the square section geometry did not affect significantly the column performances. Stationary phase films of different thicknesses, from 0.032 to 0.260 µm, were coated on different microchips of the same production batch to assess the influence of film thickness on the chromatographic performances. Microchips performance was evaluated through Golay's plots, and some of the microchips were also studied by optical microscopy. The efficiency and retention capacity of our microchip column is shown to be highly dependent on the film thickness (9,000 plates.m-1 with k = 0.12 for the thinnest film and up to almost 4,000 plates.m-1 with k = 0.93 for the thickest film). Finally, the coating process repeatability was validated by producing three identical microchips with RSD of 4.8 % for the average number of theoretical plates.

The influence of storage conditions on hygiene condition and eicosapentaenoic acid oxidation in Nannochloropsis algae for sustainable food supply.

Nannochloropsis algae contain approximately 20% polyunsaturated fatty acids (PUFA) and hold significant potential for high-quality eicosapentaenoic acid (EPA) food industrialization. However, EPA in Nannochloropsis sp. is prone to oxidation, and microbial growth is a critical factor affecting the shelf life of fresh food. Storage composition and temperature are primary factors influencing microbial growth, yet these aspects are not fully understood. This study investigates the effects of temperature and encapsulation on EPA content in nano-products over time. Nano-powder and nanobeads derived from Nannochloropsis sp. served as raw materials. Additionally, changes in aerobic plate counts and coliform groups were monitored. The results indicated that nanobeads, due to their more complex processing and less mature packaging, were more susceptible to coliform contamination compared to nano-powder. In terms of EPA stability, nanobeads exhibited a longer storage life than nano-powder. The oxidation rate of both nano-powder and nanobeads was faster at 37 °C than at 25 °C. These findings can inform general shelf life estimation, rapid detection of total lipid content in nano-products and macro extraction of nano-oil. Moreover, they have significant implications for delaying EPA oxidation in nano-products and improving hygienic quality control for microbial detection. © 2024 Society of Chemical Industry.

High-pressure processing and heat treatment of Murrah buffalo milk: Comparative study on microbial changes during refrigerated storage

This study aims to evaluate the effect of high-pressure processing (HPP) (500 and 600 MPa for 3 min and 5 min) on the microbial changes of Murrah buffalo milk in comparison to heat treatment (72 °C for 15 s of holding time) during refrigerated storage of 28 days. The results indicated that the total plate count (TPC) of raw milk at day 0 was 5.5 ± 0.6 log10 CFU/mL. At day 0, heat treatment lowered TPC to 3.9 ± 0.6, while HPP treatment was in the range of 4.1 ± 0.3 to 4.8 ± 0.6 log10 CFU/mL. Similarly, lowered yeast and mold count and lactic acid bacteria were noted in heat- and HPP-treated milk samples compared to the control sample during refrigerated storage. There were no Staphylococcus aureus and Escherichia coli detected in heat and HPP-treated samples. Heat or HPP treatment at 600 MPa for 5 min significantly extended the shelf-life of Murrah buffalo milk for three weeks at the refrigerated storage. In addition, HPP treatment did not alter the pH, lightness (L* value), protein, or fat content of Murrah buffalo milk during refrigerated storage. Hence HPP at 600 MPa for 5 min could be a suitable alternative to conventional heat treatment.

Lag Plot: A Novel Method to Determine Viable Bacterial Cell Number via a Single OD Measurement.

Bactericidal activity is a valuable parameter which is considered and measured for antimicrobial compounds. The available standard protocol to evaluate bactericidal activity is based on the direct colony count. Colony counting requires serial dilution, plating, overnight incubation, and direct counting, which is time-and labor-intensive. In regard to eliminate direct plate count, novel techniques were developed based on the real-time growth monitoring which can come with some limitations and drawbacks. These drawbacks encourage us to develop a novel technique with simple procedure to determine viable bacterial cell count. In this procedure, real-time growth monitoring is not required. In fact, after an incubation time, the number of viable bacteria can be determined with a single OD measurement and through an equation. In this regard, four standard bacterial strains, including Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), and Bacillus subtilis (ATCC 23857), were cultured with descending inoculum densities (1.5 × 107 to 15CFU/mL) and growth curves were drawn. As expected, growth in the samples with lower inoculum densities recorded with longer lag phase. Also, a direct relation was observed between the recorded turbidities and initial cell counts. A logarithmic curve (lag plot) was obtained by plotting the OD, after 12-18h incubation, against initial cell counts. In all examined strains, R2 was calculated in the range of 0.96-0.99 which is acceptable value for coefficient of determination. Equation corresponding to the lag plot was obtained and called lag equation. This equation is applicable to calculate the number of viable cells in unknown samples simply by inoculation, incubation, and single OD measurement. Developed technique can be introduced as a simple substitution for labor- and time-intensive direct colony counting.

Comparative Adhesion of Pseudomonas aeruginosa to Human Oral Mucosal Epithelial Cells and Polystyrene Surfaces

Background: The adhesion of bacteria to biotic and abiotic surfaces reflects their ability to cause infectious diseases. The distinction between the ability of Pseudomonas aeruginosa bacteria to adhere to biotic and abiotic surfaces is not clear in the literature. Objectives: The current study aims to shed light on the extent of similarities and differences between P. aeruginosa in terms of their ability to adhere to different biotic and abiotic surfaces. Materials and Methods: ten isolates of P. aeruginosa were isolated from 100 wound and burn swabs. The isolates were identified using biochemical and phenotypic tests in addition to VITIK-II technology. The susceptibility to antibiotics was estimated using the disc diffusion method. The microtiter-spectrophotometric was used to measure the biofilm formation on polystyrene microtiter plates. The Human oral mucosal epithelial (OMECs) were used to evaluate the adhesion of P. aeruginosa isolates. Plate count and direct bacterial count were used to count the adhered bacteria to human OMECs in vitro. Results: Norfloxacin showed the highest antibacterial effect while the lowest antibacterial effect was seen in the case of using when Amoxicillin and Cefixime (all isolates were resistant). All isolates from, Pa2 form the highest biofilm followed by Pa6, while the lowest biofilm formation was seen in the case of Pa4. While Pa 6 showed the highest ability to attach to human OMECs followed by Pa2. Conclusion: It can be concluded that P. aeruginosa isolates are resistant to most antibiotics and that their ability to adhere and biofilm formation depends on the type of surface.

Exposure dose, light distribution and wavelength affect the fate of introduced bacterial biological control agents in the phyllosphere of greenhouse grown tomato

Societal Impact StatementThe use of chemical plant protection products must be reduced to promote sustainability in food production. One possible alternative is biological control agents (BCAs), but their efficacy under commercial conditions does not always reach the standard of chemical control agents. Previously, light has been found to induce mechanisms in bacterial BCAs that can affect their distribution and establishment. This could promote BCA efficacy. We looked into how monochromatic and polychromatic (which is what growers use) light treatments affected the occurrence of three BCAs post‐application. By combining two non‐chemical methods: a biological (BCA) and a cultural (light) control method, this offers a new integrated pest control strategy.Summary The dynamics and functionality of beneficial and non‐beneficial, non‐phototrophic bacteria can be influenced by light quality. We investigated if light could aid the survival of three bacterial biological control agents (BCAs; Bacillus amyloliquefaciens DSM7, Pseudomonas chlororaphis 50083 and Streptomyces griseoviridis CBS904.68) in the canopy of greenhouse‐grown tomatoes at four light treatments. Tomato plants were exposed to 50 μmol m−2 s−1 of either polychromatic light (white) or monochromatic light (blue: 420 nm, green: 530 nm and red: 660 nm) using DYNA LED lamps for a total of 48 h post foliar application of the BCAs. Leaves were harvested from two levels in the canopy at the top and middle of each plant at 0, 2, 4, 8, 12, 24 and 48 h post inoculation. The occurrences of the BCAs were quantified by plate count and droplet digital PCR (ddPCR). S. griseoviridis persisted under most treatments, whereas P. chlororaphis and B. amyloliquefaciens preferred the polychromatic and green light treatments as depicted by the viable count analyses. Significant differences between the DNA and cDNA concentrations were only noted for P. chlororaphis, with prominent wavelength effects. Light exposure dose, placement in the canopy and wavelength were found to be decisive factors for BCA re‐isolation, indicating that they have different optimal light environments.

Comprehensive numerical and experimental analysis the impact of employing vortex generators on evaporative cooling process

Reduced energy usage is an important concern in vapor compression cooling systems, particularly in hot climates. These systems function poorly in hot weather and consume a lot of electricity. Evaporative condensers use the cooling impact of evaporation to facilitate the process of heat rejection and thereby increase energy efficiency. This work evaluates the impact of employing vortex generators on the performance of an evaporative condenser. The analysis is performed using a validated numerical model, a commercial CFD code. The proposed vortex generators are parallel plates with sinusoidal and zigzag patterns. Dry air enters a channel equipped with four vortex-generating plates and water spray. The incoming dry air cools due to the two-phase heat transfer during air contact with the sprayed water droplets, and the cooled air with the proper humidity level exits the channel. The investigated parameters include longitudinal nozzle position, inlet air flow rate, pitch, and the height of the vortex generator plates in two sinusoidal and zigzag modes and the obtained results are compared with the plain channel model. Greater inlet air velocity resulted in superior output temperatures and pressure drops, yet lower outlet relative humidity and water mass percentage. According to the findings, the air pressure drop and outlet dry temperature are raised by 152 % and 5 %, respectively, when the incoming air velocity is altered from 0.4 to 0.6 m. s−1 (50 % growth), and the nozzle position is moved from X = 1.35 m to X = 1.85 m (37.04 % change) from the channel entrance, while the relative humidity and water mass fraction are decreased by 19 % and 11 %, respectively. Additional research revealed that the vortex flow level is lower when the nozzle is positioned farther away from the plates, and the water spray's impact on lowering the local air temperature in vortex-flowing areas is more substantial when it is positioned closer to the plates. Furthermore, increasing wave height and pitch values of vortex generators result in a more extensive air pressure drop in the channel, and vice versa. This means that, at a fixed pitch, a higher plate wave height raises the amount of vortex flows. According to the findings, sinusoidal vortex plates have better efficiency than the zigzag ones, and in a channel equipped with water spray, the use of vortex plates before the water spray nozzles lowers the dry temperature at the outlet by about one °C compared to that of the plain channel.

An Alternative Approach to Validate the Cleaning Efficiency of a Skin Cleansing Wipe

A key functionality for cleansing wipes is their efficiency in removing dirt and microbial contamination from the skin to safe or non-detectable levels, traditionally determined using the gravimetric method, which has been reported to be prone to experimental errors. This study evaluates the efficiency of a water-based cleansing wipe, WaterWipes® (WaterWipes, UC, Drogheda, Ireland), for removing synthetic faecal matter (FecloneTM, SiliClone Creations LLC, Havertown, PA, United States) and Escherichia coli (NCTC 10538) from volunteers’ skin, the former using a dermal analytical device called the Antera 3DTM camera (Miravex Ltd., Dublin, Ireland), and the latter using standard microbiological methods. FecloneTM was applied to participants’ forearms and the Antera 3DTM camera captured detailed images of the skin surface before and after wiping. The Antera 3DTM camera approach was found to be effective in measuring cleaning efficiency, with the wipe removing all detectable traces of the FecloneTM applied. The total pore area (mm2), pore count, and total pore volume (mm3) in test participants post-wiping were observed to be reduced on average by 39.05%, 34.39%, and 39.98%, respectively. The wipe removed 99.99% of E. coli (NCTC 10538) applied, as measured using the microbial plate count method. In conclusion, the Antera 3DTM camera method was observed to be effective in evaluating removal of topically applied FecloneTM.

Microbiological profile and prevalence of histamine-producing bacteria in fresh sardines stored at different temperatures

Background and Aim: The European pilchard (Sardina pilchardus) is an important fish species for the Moroccan economy in terms of production and export. Biogenic amine histamine is a metabolite produced in the flesh of some fish species after death due to the decarboxylation of free histidine by histaminogenic bacteria. Failure to control the histamine risk in European pilchard may lead to public health and socioeconomic issues. This study aimed to estimate the prevalence of histaminogenic bacteria in association with histamine levels and the growth of microflora in Moroccan sardines (European pilchard). Materials and Methods: We conducted the study by monitoring Moroccan sardines of histamine content and microbiological profile (aerobic plate count [APC], coliforms, and thermo-tolerant coliforms [TTC]) during 6 days of storage at three different temperatures (0°C, 10°C, and ambient temperature [22°C]). The histamine assay was performed using a spectrofluorometric method, and the microbiological identification of histamine-producing bacteria was performed using a combination of biochemical and molecular tests. Results: The histamine content in European pilchard stored at 0°C was negligible. However, high concentrations were observed at 10°C and 22°C. The microbiological profile showed a positive association between microflora counts and histamine content according to storage time. At 0°C, a moderate increase in the APC, a decrease in coliforms, and an absence of TTC were observed. The rapid proliferation of all microflora was observed at 10°C, whereas at 22°C, the proliferation was almost exponential. Bacterial identification revealed the exclusive presence of species belonging to the Enterobacteriaceae family at varying frequencies depending on storage temperature. Morganella morganii and Proteus mirabilis had the highest histamine induction rates in L-histidine-supplemented broth, with 1600 and 255 parts per million (ppm), respectively, after 48-h incubation at 35°C. Klebsiella ozaenae could produce 136 ppm and Serratia plymuthica 115 ppm. Reverse transcription polymerase chain reaction showed positive results for the presence of genes associated with histidine decarboxylase. The hdc genes of M. morganii, P. mirabilis, and K. ozaenae were successfully amplified and exhibited strong similarity with the reference gene of M. morganii. Conclusion: This study describes for the first time the hdc gene in bacteria that form histamine in Moroccan sardines. The results also confirm that respect for the cold chain integrity is a crucial factor in histamine management. This information should help stakeholders in the implementation of sound strategies for managing the hazards associated with seafood and their products. Keywords: Enterobacteriaceae, histamine, Histaminogenic bacteria, polymerase chain reaction, sardine, seafood.

Multiscale simulation of liquid chromatography: Effective diffusion in macro–mesoporous beds and the B-term of the plate height equation

We performed multiscale simulations of analyte sorption and diffusion in hierarchical porosity models of monolithic silica columns for reversed-phase liquid chromatography to investigate how the mean mesopore size of the chromatographic bed and the analyte-specific interaction with the chromatographic interface influence the analyte diffusivity at various length scales. The reproduced experimental conditions comprised the retention of six analyte compounds of low to moderate solute polarity on a silica-based, endcapped, C18 stationary phase with water‒acetonitrile and water–methanol mobile phases whose elution strength was varied via the volumetric solvent ratio. Detailed information about the analyte-specific interfacial dynamics received from molecular dynamics simulations was incorporated through appropriate linker schemes into Brownian dynamics diffusion simulations in three hierarchical porosity models received from physical reconstructions of silica monoliths with a mean macropore size of 1.23 µm and mean mesopore sizes of 12.3, 21.3, or 25.7 nm. The mean mesopore size was found to have a similar influence on the effective mesopore diffusivity as the analyte polarity and the mobile-phase elution strength, which together determine the analyte residence time on a column. A smaller mesopore size attenuated the increase of the effective mesopore diffusivity with increasing mobile-phase elution strength significantly. The effective bed diffusivity was limited by the analyte residence time rather than by morphological details of the mesopore space. The stronger an analyte was retained by the chromatographic interface inside the mesopores, the slower became the mass transfer between the pore space hierarchies and the lower was the effective bed diffusivity. The B-terms of the plate height equation were finally generated with the bed diffusivities and phase-based retention factors derived from the hierarchical porosity models using additional information about the stationary-phase limit obtained from the analysis of analyte–bonded phase contacts. The B-terms highlight analyte- and mobile phase-specific behavior relevant to isocratic and gradient elution conditions in chromatographic practice. In particular, U-shaped B-term curves are observed due to the dominating contribution of the retention factor and the bed diffusivity to the B-term at low and high elution strength of the mobile phase, respectively.

THE INDUSTRIAL IMPORTANCE OF TECHNOLOGY TRANSFER FOR ANALYTICAL METHOD DEVELOPMENT AND VALIDATION–APPLICATION TO VILAZODONE HYDROCHLORIDE DOSAGE FORM

Objective: The research aims to create a fast, economical, and efficient tech transfer method for analyzing bulk materials and pharmaceutical dosage forms. Methods: The analysis was performed using an Acquity UPLC BEH C18 analytical column (130 Å, 1.7 µm, 3 mm x 100 mm) with detection of the analyte at 238 nm. The mobile phase flow rate was optimized at 0.3 ml/min, and an injection volume of 0.5 µl** was used. The isocratic mobile phase consisted of a blend of buffer (0.1% OPA) and acetonitrile at a ratio of 40:60% v/v. Results: All validation parameters of the analytical method were acceptable. The analyte eluted at 3.08 min with satisfactory theoretical plates and a tailing factor. The recovery of the analyte was 99-100%, and the method's precision was 0.59 and 0.72% RSD. The technique showed no interference, indicating specificity. The linearity was demonstrated over the 20-60 µg/ml range, with a regression coefficient of 0.999. Conclusion: The developed method is cost-effective, specific with minimal interference, and time-saving. It also reduces solvent consumption for quality control analysis of vilazodone hydrochloride samples. The method demonstrates good linearity, robustness, and accuracy.

Comparison of piperine content, antimicrobial and antioxidant activity of Piper chaba root and stem

Piper chaba (locally known as “Choi Jhal”) is used traditionally as spices and folk medicine in different parts of Bangladesh. One of the most important bioactive compounds in this plant is piperine. In this study, the amount of piperine in P. chaba root and stem was investigated and the optimal solvent for piperine extraction at room temperature was also studied. High performance liquid chromatography (HPLC) was operated using a reverse phase column where methanol and water (70:30) were used as mobile phase. The detection was performed using photo diode array (PID) detector at a wavelength of 345 nm. The standard piperine showed linearity between 0.005 % and 0.04 % and the correlation co-efficient found for the linearity was 0.9933. The percentage of relative standard deviation (RSD) for both retention time and peak area were less than 2.0 %. The theoretical plate number (N) > 3000 and a tailing factor (T) < 1.5 were found in the acceptable range. The recovery percentage (%) of standard piperine was 99.16 %. Low value of co-efficient of variation and standard deviation are recognized for high precision of the method. The highest amount of piperine was found in root extracted with methanol (MR) amounting to 1.75 % in the root powder. The maximum amount of piperine in the stem was 1.59 % when extracted with methanol (MS). The piperine quantification in other extract like n-hexane root (HR), ethyl acetate root (ER), n-hexane stem (HS), ethyl acetate stem (ES) were 0.76 %, 1.69 %, 0.33 % and 1.46 % respectively. Methanol has given the highest yield of piperine compared to ethyl acetate and n-hexane for both root and stem. The developed method was simple, rapid, economic and validated in terms precision, accuracy and recovery. This selective method is found to be repeatable, accurate and successfully utilized for the Piper extract in marketed and pharmaceutical samples with well chromatographic conditions. The ethyl acetate extract of root and stem showed promising DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging activity with an IC50 value of 39.62 ± 0.95 μg/mL and 43.85 ± 1.50 μg/mL respectively. The study reports potential antibacterial activity and antifungal activity of P. chaba root and stem extracts. These outcomes revealed that different extracts of P. chaba may be used as natural preservatives.

Microbiological and Chemical Characteristics of Yoghurt Incorporated with Watermelon Rind Powder

Watermelon rind accounts for approximately one-third of the overall fruit mass. It is usually discarded due to its low commercial value. However, it is reported to contain valuable nutrients and is an effective source of pectin that can act as a potential prebiotic. This study aimed to study the effects of watermelon rind powder (WRP) on the growth of probiotic bacteria in yoghurt and its chemical characteristics. Watermelon rind was dried by using a dehydrator and ground into powder form before being incorporated into fresh yoghurt at 2% and 4% w/v. A sample with 0% w/v WRP was prepared as control. The effect of WRP on the growth of probiotic bacteria was determined by MRS plate count. Chemical analyses including titratable acidity, pH and Brix were conducted during the fermentation process. The results showed that the increase in WRP percentage resulted in a significant increase in bacterial growth with 7.20 ± 0.22 log CFU/mL for the control sample as compared to 8.42 ± 0.23 log CFU/mL for sample with 4% WRP after 30 hr of incubation. The fermentation time was also improved with the presence of WRP with a 0.22 h-1 increase in growth rate observed for the sample with 4% WRP as compared to the control sample. Furthermore, samples containing 4% WRP showed the highest increment in titratable acidity (12.47) and the highest percentage in Brix value reduction (51.04%) during the fermentation period as compared to the control sample. Biochemical analysis showed negative values for oxidase and catalase test while positive values were obtained for gram-staining indicating the presence of Lactic acid bacteria from the gram-positive group. This study demonstrates the high potential of WRP in promoting bacterial growth for yoghurt production which is beneficial to the food industry other than promoting the ongoing effort of food waste reduction.

Synthesis of Sulfonated Chitosan as An Active Agent of Antibacterial Packaging for Fish Fillets

Modifying chitosan by increasing its hydrophilic properties through sulfonation reactions will increase its solubility in water and antibacterial activity and expand the application of chitosan. This study aims to determine the antibacterial activity of sulfonated chitosan film applied as fish fillet packaging using the total plate count method. Sulfonated chitosan in this study was successfully synthesized by reacting chitosan with 1,3-propane sultone. Sulfonated chitosan products were characterized using FTIR and UV-Vis spectrophotometers, while antibacterial activity was measured using the disc diffusion method. The optimum temperature for sulfonated chitosan synthesis was 60°C (SCS60). The resulting SCS 60 was a yellowish-white solid soluble in water with a yield of 61.46% and a degree of substitution of 33%. The diameters of the SCS 60 inhibition zone against E. coli and S. aureus isolates were 7 and 10 mm, respectively. SCS60-gelatin film is elastic (elongation 272%) and more hydrophobic with a contact angle of 90.12° compared to chitosan-gelatin film (62.8°). SCC60-gelatin film was able to suppress bacterial growth in fish fillets by up to 0.3 × 104 cfu/g compared to unpackaged fish (30 × 104 cfu/g). Sulfonated chitosan has the potential to be an antibacterial food packaging material.

Propidium monoazide (PMA) qPCR assay compared to the plate count method for quantifying the growth of Salmonella enterica serotypes in vacuum-packaged turkey breast combined with a mathematical modeling approach

This study compares the plate count (PC) and the Propidium Monoazide-quantitative Polymerase Chain Reaction (PMA-qPCR) methods to assess the growth of a cocktail of three serotypes of Salmonella enterica (Heidelberg, Typhimurium, and Enteritidis) in cooked, sliced, and vacuum-packaged turkey breast (STB) under isothermal storage temperatures (8 °C–20 °C), using predictive models. Standard curves were developed for PMA-qPCR, demonstrating high efficiency (101%) and sensitivity, with quantification limits ranging from 1 to 2 log10 CFU/g for all temperatures studied. Comparative analysis revealed a significant correlation (R2 = 0.99; 95% CI) between the PC and PMA-qPCR methods; however, the agreement analysis indicated a mean difference (Bias) of −0.11 log10 CFU/g (p < 0.05), suggesting underestimation by the PC method. This indicates the presence of stressed or viable but nonculturable (VBNC) cells, detectable by PMA-qPCR but not by PC. The Baranyi and Roberts model showed a good ability to describe the behavior of S. enterica cocktail in STB for PC and PMA-qPCR data under all isothermal conditions. The exponential secondary model more accurately represented the temperature dependence of the maximum specific growth rate compared to the Ratkowsky square root model, with R2 values ≥ 0.984 and RMSE values ≤ 0.011 for both methods. These results suggest that combining PMA-qPCR with predictive modeling allows for a more accurate prediction of S. enterica growth, compared to PC method. In the event of cold chain disruptions of meat products, the use of PMA-qPCR method allow the quantification of VBNC cells, that can still pose a health risk to consumers, especially in ready-to-eat products.

Potential Fermentation of Sweet Potato Pickle (Ipomoea batatas L.) with Lactobacillus plantarum B1765 in the Production of Short Chain Fatty Acids

Short-chain fatty acid (SCFA) is the final metabolic product of inulin that can provide various health benefits. In this study, the effect of fermentation duration of sweet potato pickle using Lactobacillus plantarum B1765 starter culture in the production of SCFA was studied, as well as its relationship with total Lactic Acid Bacteria (LAB), pH, and Total Titratable Acid (TTA). Fermentation was carried out with the addition of starter culture as much as 10% (v/v) at 37ºC for 4, 8, 12, 24, and 0 hours which was used as a control. The number of LAB was measured using the Total Plate Count (TPC) method, pH value was measured with a pH meter, TTA was measured using the acid-alkalimetric titration method, and SCFA was measured using an HPLC instrument with 0.1% H3PO4 eluent and 25% acetonitrile. Fermentation duration affects LAB growth, TTA, SCFA, and pH. The results showed that total LAB growth was optimal at 8 hours fermentation time at 2.81×108 CFU/mL, which increased by 2 log cycles from the initial fermentation time of 0 hours. Still, the pH decreased until it reached 3.23 ± 0.20, and TTA increased to 0,56 ± 0.02% until the end of fermentation at 24 hours. SCFA also increased until the end of fermentation at 24 hours, where acetic acid levels showed the highest levels (6.85 ± 0.30 mg/mL), followed by propionic acid (5.04 ± 0.20 mg/mL) and butyric acid (2.14 ± 0.10 mg/mL). Sweet potato pickles fermented with L. plantarum B1765 starter culture have met the SNI 01-3784-1995 standard and have the potential to be developed as a functional food source of SCFA, which has many health benefits.

Effect of Red Guava (Psidium guajava L.) Juice Concentration on Product Quality and Antioxidant Activity of Red Guava Synbiotic Yoghurt

Red guava juice has very strong antioxidant activity and has the potential to be processed into synbiotic functional beverage products that have health benefits. This study examined the effect of red guava juice concentration on product quality and antioxidant activity of red guava synbiotic yoghurt. Product quality tested included microbiological quality (total LAB) and chemical quality (pH value and Total Tritable Acid (TTA )value). The concentrations of red guava juice used were 0%, 10%, 20%, 30%, and 40%. Fermentation was carried out for 6 hours at 37⁰C by Lactic Acid Bacteria (LAB) from powdered mixed culture. Then organoleptic testing using the hedonic test, total LAB testing using the Total Plate Count (TPC) method, pH value testing using a pH meter, TTA value testing using the acid-base titration method, and antioxidant activity testing using the DPPH (2,2-diphenyl-1-picrylhydrazil) method. Test data were analyzed using SPSS software to determine the effect of red guava juice concentration on test results. The concentration of red guava juice in red guava synbiotic yoghurt affects the pH value, TTA value, and IC50 value as the result of antioxidant activity testing. Red guava juice concentration did not affect total LAB testing. Organoleptic assessment of red guava symbiotic yoghurt is in the range of like to like very much. The best formulation was red guava synbiotic yoghurt with 30% red guava juice concentration, which had strong antioxidant activity with an IC50 value of 59.637 ppm, pH value of 3.94, TTA value of 1.98%, and total LAB as much as 1.81x10⁸ colonies/g, so it has met the quality requirements of yoghurt according to Indonesian National Standard SNI 2981: 2009.

Study on Antioxidant Activity of Purple Sweet Potato (Ipomoea Batatas) Juice Fermented with Lactobacillus plantarum B1765

Purple sweet potatoes are a source of anthocyanins, which are antioxidants. However, the anthocyanin compounds in purple sweet potatoes are naturally bound in glycosidic form, so their potential as antioxidants is still limited. Fermentation is one method that can be used to degrade glycosidic bonds to liberate free phenolics so that it can increase the antioxidant potential of purple sweet potato. This research aims to study the growth of lactic acid bacteria (LAB), pH, total titratable acid (TAT), phenolic content, anthocyanin content, and antioxidant activity of sweet potato juice (Ipomoea batatas) fermented with Lactobacillus plantarum B1765 starter culture for 0.2, 4,6,8,10,12 and 14 hours. Total LAB was measured using the Total Plate Count (TPC) method, pH was measured using a pH meter, TAT was measured using acid-base titration, total phenolic content (TPC) was measured using the Folline-Ciocalteu method, anthocyanin content used the differential pH method and antioxidant activity was measured using the radical scavenging method 2, 2-diphenyl-1-picrylhydrazyl (DPPH) which is expressed in IC50 values. The results showed that fermentation time affected (p&lt;0.05) total LAB, pH, TAT, phenols, anthocyanin levels, and antioxidant activity in purple sweet potato juice. Total BAL increased to reach an optimum of 2.6 x 108 CFU/mL within 6 hours. However, until the end of fermentation for 14 hours, there was still a decrease in pH to 3.73, an increase in TAT reaching 0.426%, an increase in total phenolics up to 324.21 mg GAE/g, a reduction in anthocyanin levels (13.68 mg/L) and an increase in antioxidant activity with an IC50 of 47 .05 ppm which is classified as very strong. This product meets the Indonesian National Standards for fermented drinks and can potentially be a source of antioxidants.

Effect of natural plant-derived agents on the survival of the pathogenic bacteria Salmonella spp., Listeria monocytogenes in marinated chicken liver meat

This experimental study followed the survival (after inoculation of the bacteria) on non-marinated and marinated chicken liver meat (CLM) samples, with or without added antimicrobials (carvacrol; CA, garlic: G), stored at 4 and 10 °C for 14 days. A pomegranate pomace-based marinade (PPM) alone, or in combination with CA or G was used to monitor the survival/growth of Salmonella spp. and Listeria monocytogenes counts on CLM. A declining trend was recorded for both Salmonella spp. (4.1-4.4 and 3.8-4.3 log CFU/g) and Listeria monocytogenes (3.6-4.6 and 3.3-4.5 log CFU/g) populations in both marinated (absence of antimicrobials) and marinated (with antimicrobials) CLM samples, respectively, at 4 and 10 °C, on day 14 (final storage day). At 10°C, marination and antimicrobial treatments resulted in lower Salmonella spp and Listeria monocytogenes counts on CLM, compared to 4 °C (3.4-3.9 and 1.5-3.7 logs) in both marinated (absence of antimicrobials) and marinated (with antimicrobials) CLM samples, respectively, at 4 and 10 °C, on final day-14. Marination and combinations reduced the total plate count (TPC) by 0.3-0.9 and 0.2-0.6 log CFU/g, as compared to the control (non-marinated) in CLM samples (absence of inoculated bacteria) on final day-14. Concluding, it can be stated that a microbiological shelf-life extension of 9 and 11 days, as compared to the control’s shelf-life, was achieved at 4 and 10 oC, respectively. The initial pH values of the control (non-marinated) and marinated CLM samples at 4 and 10 oC, were 6.62 and 3.12, respectively. During the storage period, pH values for the control CLM were found to be higher, as compared to the marinated samples, and irrespective of storage temperature.

Phage-Modified Clear Hydrogel for Simultaneous Detection of Multiple Bacteria.

The proliferation speed of live foodborne pathogens is fast. A small number of pathogens will have a great impact on food and the environment if positive samples are not detected timely. In this study, transparent porous hydrogel stir bars, modified by two different phages (corresponding to two different bacteria (Escherichia coli and Hafnia sp)), have been developed for rapid detection of foodborne bacteria. A large number of samples can be analyzed simultaneously with a small animal live imager device to screen out the positive samples, while an adenosine triphosphate (ATP) bioluminescence sensor can be used to quantify the number of bacteria in the positive samples. The phage has good specificity and capture ability to bacteria, which makes the method highly sensitive. In addition, the use of multiple phages also enables the method to detect multiple bacteria simultaneously. The three-dimensional structure of the hydrogel allows it to modify more phages, and its transparent nature also allows the inside bioluminescence to be detected. Both can enhance the sensitivity of the detection. Finally, the reagents needed for bioluminescence, such as d-luciferin, can also be preencapsulated in the hydrogel, thus simplifying the detection step. Under the best conditions, the detection range of the method is 102-108 CFU·mL-1, and the limit of detection is 30 CFU·mL-1 within 11 min. The test results of actual samples show that there is no difference between using the method developed through this study and the traditional plate counting method, but the detection time is greatly shortened.