Optimization of Preparation Conditions for Quercetin Nanoliposomes Using Response Surface Methodology and Evaluation of Their Stability.

Quercetin is a flavonol compound with excellent biological activities. However, quercetin exhibits poor stability and solubility in water, which limits its application. In this study, quercetin nanoliposomes (QUE-NL-1) were prepared using an ultrasonic thin-film dispersion method, and the preparation conditions were optimized using response surface methodology. The optimal conditions for preparing QUE-NL-1 were as follows: an evaporation temperature of 35 °C, a drug concentration of 0.20 mg/mL, and a lipid bile ratio of 4:1. The encapsulation rate of QUE-NL-1 is (63.73 ± 2.09)%, the average particle size is 134.11 nm, and the average absolute value of the zeta potential is 37.50 and PDI = 0.24. By analyzing the storage temperature, storage time, and leakage rate of QUE-NL-1 in simulated gastrointestinal fluid, it was found that quercetin exhibits good stability after embedding and can achieve sustained release in intestinal juice. In addition, the cytotoxicity of QUE-NL-1 was not significant, and the survival rate of Caco-2 cells was >90% when the concentration range of QUE-NL-1 was 0.1-0.4 mg/mL. This study provides an efficient method for preparing QUE-NL-1 with small particle sizes, good stability, and high safety, which is of great significance for expanding the application range of quercetin.

A review of recent advances on cyanidin-3-glucoside: the biotransformation, absorption, bioactivity and applications of nano-encapsulation.

Anthocyanins are a class of dietary polyphenols that exhibit technological and bioactive-relevant properties. C3G is absorbed in its unmodified molecular form in the upper digestive tract, goes through the extended first-passage metabolism and its metabolites enter the bloodstream. The C3G metabolites possess health benefits such as antioxidant, cardio-protective, anti-inflammatory, neuroprotective, anti-cancer, anti-diabetic, and anti-thrombotic activities. However, the efficacy and distribution of C3G in the human body are restricted due to its low stability and bioaccessibility. Inspiringly, the lipid-, polysaccharide-, protein-, and nanocapsule-associated conjugates have achieved targeted delivery with enhanced bioaccessibility and controlled release. In this review, the absorption and transportation modes, decomposition and metabolism processes, functional activity mechanisms, and improved methods for enhancing the bioavailability of C3G are summarized. Moreover, the aspects of the gut microbiota regulation, C3G-mediated cytoprotection and different biocompatible materials applications are briefly discussed.

Sustainable emerging high-intensity sonication processing to enhance the protein bioactivity and bioavailability: An updated review.

High-intensity ultrasound (HIU) is considered one of the promising non-chemical eco-friendly techniques used in food processing. Recently (HIU) is known to enhance food quality, extraction of bioactive compounds and formulation of emulsions. Various foods are treated with ultrasound, including fats, bioactive compounds, and proteins. Regarding proteins, HIU induces acoustic cavitation and bubble formation, causing the unfolding and exposure of hydrophobic regions, resulting in functional, bioactive, and structural enhancement. This review briefly portrays the impact of HIU on the bioavailability and bioactive properties of proteins; the effect of HIU on protein allergenicity and anti-nutritional factors has also been discussed. HIU can enhance bioavailability and bioactive attributes in plants and animal-based proteins, such as antioxidant activity, antimicrobial activity, and peptide release. Moreover, numerous studies revealed that HIU treatment could enhance functional properties, increase the release of short-chain peptides, and decrease allergenicity. HIU could replace the chemical and heat treatments used to enhance protein bioactivity and digestibility; however, its applications are still on research and small scale, and its usage in industries is yet to be implemented.

[Progress on three-dimensional cell culture technology and their application].

Three-dimensional (3D) cell culture model is a system that co-culture carriers with 3D structural materials and different types of cells in vitro to simulate the microenvironment in vivo. This novel cell culture model has been proved to be close to the natural system in vivo. In the process of cell attachment, migration, mitosis and apoptosis, it could produce biological reactions different from that of monolayer cell culture. Therefore, it can be used as an ideal model to evaluate the dynamic pharmacological effects of active substances and the metastasis process of cancer cells. This paper compared and analyzed the different characteristics of cell growth and development under two-dimensional (2D) and 3D model culture and introduced the establishment method of 3D cell model. The application progress of 3D cell culture technology in tumor model and intestinal absorption model was summarized. Finally, the application prospect of 3D cell model in the evaluation and screening of active substance was revealed. This review is expected to provide reference for the development and application of new 3D cell culture models.

A Comparative Study of Pickled Salted Eggs by Positive and Negative Pressure-Ultrasonic Method.

In this study, the positive and negative pressure-ultrasonic method was applied to salted egg pickling, compared with traditional pickled salted eggs by various physical and chemical indicators. Results indicated the salt content of egg white and egg yolk increased rapidly in the salt-preserved salted egg with the positive and negative pressure-ultrasonic method, and the moisture content decreased rapidly. In addition, the oil yield of egg yolk was marinated for 12 days compared with the normal method of 35 days, and the ripening time of salted eggs was shortened by 2/3. There was no obvious difference in the microscopical structure of the egg yolk between the two methods of pickling. Moreover, the pores on the eggshell of the salted egg that was marinated by the positive and negative pressure-ultrasonic method had big cracks, which was beneficial to the substance exchange of the eggs and the outside. The common volatile flavor substances were detected by GC-MS, and a total of 33 flavor constituents were detected. There was no significant difference between the content of alcohols, aldehydes, and ketones, which contributed greatly to the flavor. Overall, the results indicated that this innovative salted eggs method can significantly reduce the curing time while ensuring the quality of salted eggs.

Protective Effect of Anthocyanins against Neurodegenerative Diseases through the Microbial-Intestinal-Brain Axis: A Critical Review.

With the increase in human mean age, the prevalence of neurodegenerative diseases (NDs) also rises. This negatively affects mental and physiological health. In recent years, evidence has revealed that anthocyanins could regulate the functioning of the central nervous system (CNS) through the microbiome-gut-brain axis, which provides a new perspective for treating NDs. In this review, the protective effects and mechanisms of anthocyanins against NDs are summarized, especially the interaction between anthocyanins and the intestinal microbiota, and the microbial-intestinal-brain axis system is comprehensively discussed. Moreover, anthocyanins achieve the therapeutic purpose of NDs by regulating intestinal microflora and certain metabolites (protocateic acid, vanillic acid, etc.). In particular, the inhibitory effect of tryptophan metabolism on some neurotransmitters and the induction of blood-brain barrier permeability by butyrate production has a preventive effect on NDs. Overall, it is suggested that microbial-intestinal-brain axis may be a novel mechanism for the protective effect of anthocyanins against NDs.

Anti-Allergic Effects of Quercetin and Quercetin Liposomes in RBL-2H3 Cells.

BACKGROUND: Quercetin is a kind of flavonoid with important bioactivities, such as hypoglycemic, antioxidant, anti-inflammatory, and anti-allergic properties. Although it is unstable, it is worth exploring how to better exert its anti-allergic effect. OBJECTIVE: The current study aimed to elucidate the anti-allergic effect of quercetin liposomes on RBL-2H3 cells in vitro. METHODS: Quercetin liposomes were prepared to improve the anti-allergic activity of quercetin through a green thin-film dispersion method. We compared the anti-allergic effects of quercetin and quercetin liposomes in RBL-2H3 cells. The anti-allergic activity of the quercetin liposomes was evaluated by the level of ß-hexosaminidase, histamine, Ca2+, IL-4, IL-8, and MCP-1. RESULTS: The results showed that quercetin liposomes could significantly restrain the release of ß-hexosaminidase and histamine, calcium influx, and the expression of inflammatory factors, whose effect is stronger than quercetin. CONCLUSION: Collectively, our research suggests that the quercetin liposome can be used as a potential allergy antagonist.

A review on the protective effect of active components in Antrodia camphorata against alcoholic liver injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Antrodia camphorata is a genus of wood-rot basidiomycete in the family Fomitopsidaceae. It is a valuable medicinal fungus in China that contains more than 78 kinds of active compounds. A. camphorata has good protection effects on the liver, especially on alcoholic liver injury (ALI). AIM: This paper summarizes the complex occurrence and development of alcoholic liver disease (ALD). In addition, the effect of ALD on the intestine through the gut-liver axis is summarized. The protective mechanism of A. camphorata on ALI is reviewed to reveal its therapeutic potential, offering insights into future research. MATERIALS AND METHODS: A comprehensive search in the literature was obtained from books and online databases such as Web of Science, Google Scholar, PubMed, Scopus, Science direct, ACS Publications and Baidu Scholar. RESULTS: The pathogenesis of ALD mainly includes oxidative stress injury, intestinal microflora imbalance, inflammatory mediator injury and nutritional imbalance. A. camphorata contains rich active components (e.g. polysaccharides, triterpenoids, maleic and succinic acid derivatives, amino acids, superoxide dismutase, vitamins, lignin and sterols). These components have good antioxidant, anti-inflammatory and intestinal protection activities. Therefore, A. camphorata has a wide application in the prevention and treatment of ALI. CONCLUSIONS: ALD develops from a mild disease to alcoholic hepatitis and cirrhosis, which is the main reason of global morbidity and mortality. At present, there is no effective drug for the treatment of ALD. A. camphorata, as a valuable medicinal fungus unique to Taiwan, has a great protective effect on the liver. It is expected to be an effective drug for ALI treatment. Although many studies have performed the protective effects of A. camphorata on ALI, its regulatory effects on the gut-liver axis of ALD patients need to be further explored.

Use of different endpoints to determine the bioavailability of polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and polychlorinated biphenyls (PCBs) in Sprague-Dawley rats.

Liver, fat (adipose tissue), blood, and feces are common endpoints used to determine the bioavailability of persistent organic pollutants (POPs). However, it is not known whether the bioavailability of each endpoints is comparable or whether there is a comprehensive endpoint that can be used for all congeners for the measurement of bioavailability. In this study, we observed the accumulation and distribution of 10 polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and 18 polychlorinated biphenyls (PCBs) in different organs of Sprague-Dawley (SD) rats and calculated the bioavailability based on feces, liver, and fat endpoints. Our results indicated that PCB 126, PCB 169, and 50% of PCDD/F congeners were mainly accumulated in the liver, with a bioavailability ranging from 28.9 to 50.6%. On the other hand, higher chlorinated (> 5 Cl) PCB congeners were mainly accumulated in adipose tissues, with a bioavailability ranging from 20.1 to 82.2%, while lower chlorinated (< 5 Cl) pollutants, such as 2,3,7,8-TeCDF, 2,3,7,8-TeCDD, 1,2,3,7,8-PeCDF, and PCB 28, 52, 77, 81, were likely metabolized over 36% in rats during the 8-week experimental period. If we considered metabolization (degradation) as a type of bioavailable process, then the fecal endpoint was a feasible option. However, if we considered the selective accumulation behavior of some congeners in different organs/tissues, then there was no single comprehensive endpoint suitable for all congeners. Lastly, female rats showed significantly higher PCDD bioavailability than male rats at low dose level (0.2 ng/100 g b.w./d); however, the difference in PCB bioavailability between female and male rats was not significant.

Cellular uptake, transport mechanism and anti-inflammatory effect of cyanidin-3-glucoside nanoliposomes in Caco-2/RAW 264.7 co-culture model.

Cyanidin-3-glucoside (C3G), which is the widest and richest anthocyanin (ACN) found in the edible fruit and vegetables, has been illustrated to perform a wide range of bioactivities. Nanoliposomes can inhibit C3G degradation and enhance the absorption rate of C3G as tools for conveying materials to particular locations. This experiment aims to study the absorption, transport and anti-inflammatory effects of C3G nanoliposomes in Caco-2/RAW 264.7 co-culture model, which symbolizes an intestinal inflammation system. The results indicated that the uptake and transport of C3G nanoliposomes by Caco-2/RAW 264.7 co-culture model were concentration-dependent as well as affected by temperature (37 and 4°C) and endocytic inhibitors, which revealed C3G nanoliposomes penetrate cells via endocytosis. Moreover, compared with C3G, C3G nanoliposomes significantly decreased pro-inflammatory cytokine expression (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, IL-8), suggesting a stronger anti-inflammatory potential. Conclusively, the uptake of C3G nanoliposomes by Caco-2/RAW 264.7 co-culture model is mainly involved in macropinocytosis and endocytosis mediated by carrier protein (clathrin). C3G nanoliposomes may play a better role in the treatment of LPS-induced intestinal inflammation diseases.

Solid-state-cultured mycelium of Antrodia camphorata exerts potential neuroprotective activities against 6-hydroxydopamine-induced toxicity in PC12 cells.

Antrodia camphorata (A. camphorata) is an edible fungus containing various bioactive compounds generally used for health benefits. This study aimed to explore the potential neuroprotective activities of solid-state-cultured mycelium of A. camphorata (SCMAC) against Parkinson's disease (PD), as well as the underlying mechanism using an in vitro 6-hydroxydopamine (6-OHDA)-induced PC12 cell model. The results showed that SCMAC extracts alleviated cell toxicity induced by 6-OHDA and the loss of dopaminergic neurons, which was confirmed by the increase of cell viabilities, inhibition of cell apoptosis, the upregulation of tyrosine hydroxylase (TH) and dopamine transporter (DAT) levels and the downregulation of α-Synuclein level. After purification, 11 compounds were identified by the NMR technique, including a quinone, four phenolic acid derivatives, three ubiquinone derivatives, two alkaloids, and a triterpenoid. The present study suggests that SCMAC could be an attractive candidate for the prevention or treatment of PD. PRACTICAL APPLICATIONS: Parkinson's disease seriously affects the lifetime and quality of the elder population for a long history. Long-term consumption of L-DOPA will result in side effects, such as developing abnormal involuntary movements called dyskinesia. This study showed that natural SCMAC extracts could be a potential therapeutic agent for the treatment of neurodegenerative disorder.

Soft-Shelled Turtle Peptide Supplementation Modifies Energy Metabolism and Oxidative Stress, Enhances Exercise Endurance, and Decreases Physical Fatigue in Mice.

The potential of soft-shelled turtle peptides (STP) against fatigue was evaluated. Mice orally supplemented with STP significantly increased the swimming time until tiredness by 35.4-57.1%. Although not statistically significant, STP increased muscle and thymus mass. In addition, the serum lactate, ammonia, blood urea nitrogen content and creatine kinase activity in STP-fed mice were dramatically decreased when compared to the control group. Furthermore, STP supplementation increased the reserves of liver glycogen and muscle glycogen, thus improved the energy metabolism system of mice. STP treatment contributed to increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities as well as a decrease in malondialdehyde (MDA), indicating an improvement in oxidative stress protection. The Western blot (WB) results indicated that the STP supplement effectively altered the expression of oxidative stress-related protein by modulating the NRF2/KEAP1 pathway. In summary, STP affected NRF2/KEAP1 levels in skeletal muscle, leading to antioxidant activity and a slower time to exhaustion during exercise.

Anti-inflammatory activity of cyanidin-3-O-glucoside and cyanidin-3-O-glucoside liposomes in THP-1 macrophages.

Cyanidin-3-O-glucoside (C3G) is a kind of water-soluble pigment widely existing in many plants. It has strong antioxidant and anti-inflammatory activities. However, C3G cannot exist stably for a long time because of the phenolic hydroxyl groups in its structure. Liposome technology could improve the stability and bioavailability of compounds. Based on our previous studies, C3G liposomes prepared by ethanol injection method have a certain stability in two weeks of storage. In this study, THP-1 macrophages treated with C3G and C3G liposomes can reduce the levels of inflammatory-related factors, such as tumor necrosis factor-a (TNF-a), interleukin (IL)-1ß, IL-6, and IL-8, stimulated by lipopolysaccharide (LPS). Further studies showed that the LPS induction could increase the level of phosphorylated nuclear transcription factor NF-κB and phosphorylated IkBa, while C3G and C3G liposomes could inhibit the expression of phosphorylated proteins. Moreover, C3G and C3G liposomes could protect macrophages from apoptosis. In conclusion, C3G prepared by liposome technology exhibits anti-inflammatory activity, which provides a theoretical basis for the food industry to study functional food.

Lactobacillus plantarum ZJUFB2 Prevents High Fat Diet-Induced Insulin Resistance in Association With Modulation of the Gut Microbiota.

Lactobacillus plantarum ZJUFB2 is a novel probiotic isolate derived from Chinese traditional sourdough that possesses promising probiotics properties. This study aimed to investigate the effects of L. plantarum ZJUFB2 (B2) on insulin sensitivity using mice fed on a high-fat diet (HFD) as well as to explore the involved mechanisms. Purposely, male C57BL/6 mice continuously received an intervention of B2 (~109 CFU/day) for 16 weeks. The results showed that B2 treatment remarkably ameliorated insulin resistance and hyperglycemia in HFD-fed mice. The B2 intervention significantly decreased the hepatic lipid accumulation, serum low-density lipoproteins cholesterol, and lipopolysaccharides, and regulated the bile acids levels as well as liver mRNA expression involved in lipid metabolism. Moreover, the B2 intervention significantly changed the gut microbiota, specifically, showed a lower abundance of obesity-related and inflammation-associated microbes, e.g., Ruminococcus and Mogibacteriaceae. Furthermore, it exhibited a higher abundance of short-chain fatty acids and bile salt hydrolas-producing bacteria, such as Bifidobacterium and F16 compared with the HFD group. The findings of this study suggested that B2 is a novel probiotic, effective in preventing insulin resistance by improving the gut microbiota and bile acids.

Advances in research on calf rennet substitutes and their effects on cheese quality.

Milk coagulation is an important step in cheese production, and milk-clotting enzymes (MCEs) play a major role in this process. Calf rennet is the most widely used MCE in the cheese industry. The use of calf rennet substitutes is becoming necessary due to the limited availability of calf rennet and the increase in cheese consumption. The objective of this review is to summarize the latest findings on calf rennet substitutes (animal MCEs, plant-derived MCEs, recombinant MCEs and microbial MCEs) and their application in cheese production. Special emphasis has been placed on aspects of the effects of these substitutes on hydrolysis, functional peptides, cheese variety and cheese yield. The advantages and disadvantages of different calf rennet substitutes are discussed, in which microbial MCEs have the advantages of less expensive production, greater biochemical diversity, easier genetic modification, etc. In particular, some of these MCEs have suitable characteristics for cheese production and are considered to be the most potential calf rennet substitutes. Moreover, challenges and future perspectives are presented to provide inspiration for the development of excellent calf rennet substitutes.

Anti-Inflammatory Properties In Vitro and Hypoglycaemic Effects of Phenolics from Cultivated Fruit Body of Phellinus baumii in Type 2 Diabetic Mice.

Dietary intervention in type 2 diabetes mellitus (T2DM) is a hotspot in international research because of potential threats to human health. Phellinus baumii, a wild fungus traditionally used as a food and medicine source, is now cultivated in certain East Asian countries, and is rich in polyphenols, which are effective anti-inflammatory ingredients useful in treatment of T2DM, with fewer side effects than drugs. To examine the hypoglycaemic effects of Phellinus baumii phenolics (PPE), the metabolite profiles of T2DM mice induced by streptozotocin after PPE intervention were systematically analyzed. Here, 10 normal mice were given normal saline as control group, and 50 model mice were randomly assigned to five groups and daily intragastric administrated with saline, metformin (100 mg/kg), and PPE (50, 100, 150 mg/kg of body weight), for 60 days. The pro-inflammatory factor contents of lipopolysaccharide stimulation of RAW 264.7 cells were decreased in a dose-dependent manner after PPE treatment, we propose that PPE could exert anti-inflammatory properties. PPE could also effectively reduce blood glucose levels, increased insulin sensitivity, and improved other glucolipid metabolism. Q-PCR results suggested that the hypoglycemic effects of PPE might be through activating IRS1/PI3K/AKT pathway in diabetic mice. These results suggest that PPE has strong potential as dietary components in the prevention or management of T2DM.

Separation, characterization and hypoglycemic activity in vitro evaluation of a low molecular weight heteropolysaccharide from the fruiting body of Phellinus pini.

Edible mushrooms have potential in anti-diabetic phytotherapy. They are rich in natural compounds such as polysaccharides, which have been known to have antihyperlipidemic effects since ancient times. A polysaccharide fraction of PP80 and a contained low molecular-weight (Mw), water-soluble polysaccharide (PPW-1, Mw: 3.2 kDa) were isolated from the fruiting body of Phellinus pini. Both PP80 and PPW-1 possess α-glucosidase inhibition and glucose consumption amelioration in an insulin-resistant HepG2 cell model. The α-glucosidase inhibitory activity of PPW-1 (IC50 = 2.2 ± 0.1 mg mL-1) is significantly (P < 0.01) higher than those of PP80 (IC50 = 13.1 ± 0.5 mg mL-1) and acarbose (IC50 = 4.3 ± 0.2 mg mL-1), behaving in a non-competitive inhibition manner. The structural characterization results indicated that PPW-1 is a homogeneous heteropolysaccharide composed of d-glucose, d-mannose, d-galactose and l-rhamnose. The major backbone of PPW-1 is primarily comprised of 1,6-linked glucopyranose, every third residue of which is branched at the O-3 position by a side chain consisting of 1,3-linked and terminal glucopyranose. In addition, small amounts of 1,2-linked-α-d-Manp, 1,6-linked-3-O-Me-α-d-Galp and rhamnose exist in PPW-1. In summary, PPW-1 is a novel heteropolysaccharide with potent in vitro hypoglycemic activity, and it may be a potential dietary component for improving glucose homeostasis.

Effects of ultrasonic pre-treatment on physicochemical properties of proteins extracted from cold-pressed sesame cake.

Sesame is an oil crop with high nutritional value. Protein is one of the main ingredients of sesame, however research on protein of cold-pressed sesame cake is limited. This study aimed to investigate the effects of ultrasonic pre-treatment (UPT) on physicochemical properties of proteins (yield, solubility, amino acid composition, surface properties, structural and thermal stability) extracted from the cold-pressed sesame cake, after removing lignans by ultrasonic-assisted extraction. By comparison, the extraction yield of protein was significantly (p ＜ 0.05) increased from 22.24% (without UPT) to 25.95% (with UPT), while the purity (54.08% without UPT, 55.43% with UPT), total amount of essential amino acids (22.48% without UPT, 23.10% with UPT) and non-essential amino acids (37.48% without UPT, 36.54% with UPT) were not significantly influenced. Besides, UPT slightly reduced the solubility, foaming capacity and stability (FC and FS) of protein. In addition, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR) and thermal stability (TG) analysis demonstrated that UPT could disorder and loose protein molecular structure, resulting in the change of morphology, secondary structure and thermal stability. In conclusion, this study provides a way for the separation and future application of sesame cake protein. UPT is a good option to remove the lignans from sesame cake proteins.

Using power ultrasound to release glycosidically bound volatiles from orange juice: A new method.

Acid hydrolysis and enzymatic hydrolysis are the main methods for releasing glycosidically bound volatiles (GBV). However, acid hydrolysis yields a strong pungent odor, and enzymatic hydrolysis is time consuming. In the present study, a new method, ultrasound hydrolysis, is reported to release GBV. This method is simple, environmentally friendly, fast and effective. Large differences were observed in the released aglycones and glycosyls between ultrasound and enzymatic hydrolysis of GBV. More types of aglycones were released under ultrasound than enzymatic hydrolysis. Alcohols and esters were the main aglycones under enzymatic hydrolysis, and terpenoids, esters and aldehydes were the main aglycones under ultrasound hydrolysis. The glycosyls released under ultrasound hydrolysis were mannose, glucose and sucrose, and those released under enzymatic hydrolysis were galactose and sucrose. The present study gives a new insight into a hydrolytic method for GBV by using ultrasound hydrolysis and can provide a reference method for fruit juice aromatization.

Effects of Different Smoking Materials and Methods on the Quality of Chinese Traditional Bacon (Larou).

ABSTRACT: Larou is a traditional smoked meat product in China. In this experiment, larou was processed with different smoking materials and methods to determine whether differences in processing methods would affect the quality of the larou and the concentrations of carcinogens. Pork bellies were marinated, dried, and divided into four groups and then directly smoked with four different smoking materials for 40 min. The smoking material for larou that was most effective was then used with an indirect smoking device with an nano-activated carbon fiber filter and evaluated as a single-factor variable. The surface area of the nano-activated carbon filter was 978.00 m2/g, and this filter effectively adsorbed the ash particles from the smoke. For the group smoked with pomelo skins (PS), the highest concentrations and number of phenols were 4.48% and 11, respectively, which increased the smoke flavor significantly. The moisture was 32.64%, and the Staphylococcus, lactic acid bacteria, and yeast and mold levels were 0.98, 1.10, and 0.59 log CFU/g, indicating inhibition of harmful bacteria and a beneficial microbial environment for larou fermentation. The benzo[a]pyrene (B[a]P) concentration in PS smoke determined with the indirect smoking device was 1.82 µg/kg, whereas that determined with the direct smoking device was 36.1 µg/kg, a significant difference (P < 0.01). These findings suggested that indirect smoking with PS could effectively maintain microbial quality and reduce the B[a]P[mc] concentrations in larou. This processing method can be used for the production of this meat product.