Perception of Malaysian consumers towards probiotics in fermented foods and their benefits to human health

Consumers demonstrated an increase in awareness and interest in food that are closely associated with health benefits. As such, consumers are interested in consuming probiotic products and foods. Probiotics are bacteria that offer various benefits to human health when consumed in sufficient quantities. Numerous probiotic products have been listed, including fermented foods that contain probiotics that are beneficial to human health. Hence, consumers should be aware of these products. This study aimed to analyze the knowledge and awareness of consumers about probiotics and their benefits to human health and to evaluate the perception of consumers towards probiotics in fermented foods based on the socio-demographic profiles. The questionnaire was developed using Google Forms and distributed through social media. The data was analyzed using SPSS software. 150 respondents were involved, but only 133 of the data were selected. Descriptive analysis was conducted to determine the frequency of data from the socio-demographic profiles. Meanwhile, statistical analyses using chi-square analysis was conducted to identify significant difference at p-value < 0.05 between consumers’ socio-demographic profile and their awareness, knowledge, and perceptions of probiotics and probiotics in fermented food. Overall, the survey indicated that consumers were well aware of, and had the knowledge and good perceptions towards probiotics and their presence in fermented foods

Identification and characterization of the lactic acid bacteria isolated from Malaysian fermented fish (Pekasam)

Recently researchers are interested with the biotherapeutic potential of probiotics in gut disease treatment. The bacteria are generally regarded as a safe, have a stability of usage and originate from the natural resources. The study aims to identify and characterize the potential probiotic Lactic Acid Bacteria (LAB) isolated from Malaysian fermented fish product known as Pekasam. Fourty isolates obtained were firstly screened for their antagonism activities against the common pathogenic bacteria; Esherichia coli, Staphylococcus aureus and Klebsiella sp. Our study revealed only three (labeled as L8, L20 and S1) of the isolates tested showed broad antimicrobial effects towards the pathogenic bacteria. All of the isolates were also γ-hemolytic and tolerant to various pH (pH 3, 5 and 7.5) and 0.3% (w/v) bile salts. The bacteria isolates of strain L8 and L20 were susceptible to seven antibiotics tested except vancomycin and tetracycline whereas S1 was resistant to all antibiotics. Phenotypic tests revealed that both bacteria isolates of strain L8 and L20 were Bacillus megaterium while S1 was Pediococcus pentosaceus whereas 16S rRNA gene sequence analysis showed potential bacteria isolates of strain L8 and L20 belonged to the Lactobacillus plantarum (99% similarity) and S1 was characterized as Lactobacillus pentosus (100% similarity) respectively. Our present study showed that the probiotics of strain L8, L20 and S1 isolated from the fermented fish (Pekasam) exhibited the potential probiotic properties to be developed as biotherapeutic agents

Species identification of potential probiotic lactic acid bacteria isolated from Malaysian fermented food based on 16S ribosomal RNA (16S rRNA) and internal transcribed spacer (ITS) sequences

Taxonomic identification of potential probiotic lactic acid bacteria (LAB) in fermented foods is essential as the microorganisms’ benefits are varied and often strain-specific. Using biochemical and physiological methods alone is inadequate to precisely distinguish each strain. In this study, molecular techniques were employed in the identification of 17 LAB isolated from three Malaysian fermented foods: belacan (BE), bosou (BO), and budu (BUM). The respective sizes of polymerase chain reaction (PCR) products from the isolates were approximately 1500 bp and 750 bp when amplified with the 16S ribosomal RNA (16S rRNA) and internal transcribed spacer (ITS) gene primers. The phylogenetic analysis using both gene sequences revealed that all BE and BO isolates were identified as Lactiplantibacillus plantarum, while all BUM isolates were identified as Lacticaseibacillus paracasei. Both 16S rRNA and ITS genes could disclose the identity of the isolates up to the species level. In summary, the use of the ITS gene in conjunction with the 16S rRNA gene can help with the more effective identification of potential probiotic LAB strains isolated from fermented food

Evaluation of cells integrity using different fixation time by scanning electron microscopy

The study aims to evaluate the integrity and structural changes of two intestinal cells adhered with Lactobacillus sp. in different fixation time. HT-29 and CCD-18Co intestinal cells with Lactobacillus sp. were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer for two, four, six and 12 hours at room temperature. Our study revealed that the different fixation time affects and change the integrity, viability and durability of cells. HT-29 cells structure remain intact even after 12 hours fixation while CCD-18Co cells remain intact at two and four hour fixation time whereas at six hour cells also remain intact except cilia structure not clearly seen on surface of the cell. However, at 12 hours fixation, CCD-18Co cells were completely broken and degraded. Lactobacillus sp. cells remains stable in both cell lines, showing that the bacteria cells were unaffected with various fixation time. The present study suggested fixation time is important as an aware different cell has different endurance and structural integrity

The synergistic Lactobacillus plantarum L20 and Sargassum polycystum -added diet for improvement of Black tiger shrimp, Penaeus monodon ‘s growth, immune responses, bacterial profiles, and resistance against Vibrio parahaemolyticus associated Acute hepatopancreatic necrosis disease (AHPND) infection

Acute hepatopancreatic necrosis disease (AHPND) has inflicted massive economic losses and posed a considerable threat to the development of penaeid shrimp aquaculture in Malaysia. The restriction on antibiotic uses have necessitated exploring alternatives preventive measures against AHPND outbreak, such as the use beneficial bacteria and nutritional additives. In this study, diets which were fed to Penaeus monodon postlarvae (PL15) were fed with diets containing probiotic (L. plantarum at 108 CFU/mL), prebiotic (S. polycystum at 2) and synbiotic diets (combination of probiotic and prebiotic) formulations. These diets were administered in triplicate for 35 days, followed by an immersion challenge with 6 × 105 CFU/mL AHPND-causing V. parahaemolyticus S2–4. Sampling of shrimp's cephalothorax and abdomen from the feeding trial and the immersion challenge were conducted for gene expression and histopathology analyses. Results indicated that the synbiotic-fed group displayed the most significant weight gain, specific growth rate, and protein conversion ratio among the tested groups. In addition, shrimp cephalothorax from this group displayed a significant immune response, with expression of LGBP, peroxinectin, prophenoloxidase during post-feeding trial, and expression of prophenoloxidase, toll-like receptor, penaeidin, during post-challenge trial. Furthermore, the highest L. plantarum concentration in abdomen, lowest V. parahaemolyticus S2–4 concentration in cephalothorax and highest percent survival of shrimp during post-challenge were observed in this group. Therefore, this study highlighted the positive effect of synbiotic-supplemented diet on growth, immune response, and disease resistance of shrimp, offering a promising and sustainable solution to alleviate substantial production losses in shrimp farming. © 2023 The Author

The synergistic Lactobacillus plantarum L20 and Sargassum polycystum -added diet for improvement of Black tiger shrimp, Penaeus monodon ‘s growth, immune responses, bacterial profiles, and resistance against Vibrio parahaemolyticus associated Acute hepatopancreatic necrosis disease (AHPND) infection

Acute hepatopancreatic necrosis disease (AHPND) has inflicted massive economic losses and posed a considerable threat to the development of penaeid shrimp aquaculture in Malaysia. The restriction on antibiotic uses have necessitated exploring alternatives preventive measures against AHPND outbreak, such as the use beneficial bacteria and nutritional additives. In this study, diets which were fed to Penaeus monodon postlarvae (PL15) were fed with diets containing probiotic (L. plantarum at 108 CFU/mL), prebiotic (S. polycystum at 2%) and synbiotic diets (combination of probiotic and prebiotic) formulations. These diets were administered in triplicate for 35 days, followed by an immersion challenge with 6 × 105 CFU/mL AHPND-causing V. parahaemolyticus S2–4. Sampling of shrimp’s cephalothorax and abdomen from the feeding trial and the immersion challenge were conducted for gene expression and histopathology analyses. Results indicated that the synbiotic-fed group displayed the most significant weight gain, specific growth rate, and protein conversion ratio among the tested groups. In addition, shrimp cephalothorax from this group displayed a significant immune response, with expression of LGBP, peroxinectin, prophenoloxidase during post-feeding trial, and expression of prophenoloxidase, toll-like receptor, penaeidin, during post-challenge trial. Furthermore, the highest L. plantarum concentration in abdomen, lowest V. parahaemolyticus S2–4 concentration in cephalothorax and highest percent survival of shrimp during post-challenge were observed in this group. Therefore, this study highlighted the positive effect of synbiotic-supplemented diet on growth, immune response, and disease resistance of shrimp, offering a promising and sustainable solution to alleviate substantial production losses in shrimp farming

In vivo assessment of Lactobacillus plantarum strains in black tiger shrimp (Penaeus monodon): implications for growth performance, probiotic-pathogen interaction, and defense against AHPND infection

Acute hepatopancreatic necrosis disease (AHPND) is an emerging disease that has recently severely damaged global shrimp aquaculture industries. Aside from antibiotics, using a probiotic, Lactobacillus spp., can become an alternative for AHPND prevention in shrimp aquaculture. Therefore, this study aims to screen the most suitable Lactobacillus strains for black tiger shrimp (Penaeus monodon) against AHPND. Forty-nine lactic acid bacteria were identified out of 232 isolates from fresh wild-caught fish and shrimp in the local wet market. From there, six Lactobacillus spp. strains were chosen for the hemolytic test, colony top assay, well diffusion assay, and co-culture assay. Subsequently, three strains of L. plantarum, including strains L20, KD2, and T31, were chosen for feeding trial. Bacterial dosages of 108 CFU/mL of the L. plantarum were incorporated into basal feed, respectively. A total of 600 shrimp postlarvae (PL15) were distributed into 4 treatment groups (control, L20, KD2, and T31) with triplicates. Shrimp in treatment groups were fed with different supplemented feeds for 35 days. On the 35th day, all shrimp were challenged via immersion with 6 × 105 CFU/mL of Vibrio parahaemolyticus S2-4 positive AHPND. All shrimp from the feeding trial and the challenge test were sampled for gene expression and histopathology throughout the study. Results revealed that L. plantarum strain L20 was the best probiotic among the three strains since it improved shrimp growth performance during the feeding trial, shrimp gastrointestinal microbiota, and survival of P. monodon postlarvae against V. parahaemolyticus S2-4 in the challenge test. This study demonstrated the potential allochthonous L. plantarum L20 as a probiotic in shrimp against AHPND, which causes significant losses in the aquaculture industry of Malaysia