Many studies have shown that probiotic strains added to a number of probiotic products are not compatible to that of claimed. It is thus of note to validate probiotic strains added to probiotic products. In this study, three probiotic drinks, A, B and C, were cultured on MRS agar and the number of bacterial colonies was enumerated. The bacterial counts recovered from A (9.3 ± 6.9 log CFU/ml) and C (9.0 ± 6.9 log CFU/ml) were signifi cantly higher than B (5.2 ± 3.5 log CFU/ml) and achieved the minimal amount recommended for probiotic bacteria. All of the isolates appeared as gram positive rods microscopically and were proven to be catalase negative. However, there were only A1, A2, B4 and C1 that were highly tolerant to the gastrointestinal pH 3 to 6. The four isolates produced and secreted antimicrobial substances which inhibited the growth of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). C1 showed the greatest growth inhibition by forming 17.50-mm and 17.85-mm inhibition zones against E. coli and S. aureus, respectively. The 16s rDNA sequencing and phylogenetic analysis were performed to further identify the twelve isolates. The twelve isolates were found to be Lactobacillus (L.), particularly L. casei and L. paracasei. However, the bacteria isolated from drink B were incompatible to the labelled ones. In conclusion, probiotic drinks are possible to contain different bacterial counts and probiotic strains from the labelled ones. These differences might affect health benefi ts rendered by probiotic strains to consumers.
Probiotics

The occasional influenza pandemics and the seasonal influenza epidemics have destroyed millions of lives since the last century. It is therefore necessary to understand the virus replication patterns as this provides essential information on the virus infectivity, pathogenicity and spread patterns. This study aimed to investigate the replication of avian influenza A virus H5N1 (A/Chicken/Malaysia/5858/2004) in MDCK cells. In this study, the TCID50 (50% tissue culture infectious dose) of AIV H5N1 was first determined. The MDCK cells were then infected with AIV H5N1 at TCID50 for 0-48 h. The CPE (cytopathic effect) was observed and cell death was determined hourly. The virus-infected cells and media were subsequently collected for gene analysis. The results showed that the TCID50 of AIV H5N1 was 10-9 dilution. The CPE percentage showed a strong and positive correlation with the infection period (r = 1.0, n = 9, p < 0.01). The amount of a highly conserved influenza viral gene, M2 gene amplified from infected media (r = 0.471, n = 9, p= > 0.05) and infected cell (r = 0.73, n = 9, p < 0.05) were also positively correlated with the infection period. In conclusion, although CPE started to be observed in the early time points of infection, however, the M2 gene was only amplified from the infected media and cells after 48 h and 24 h, respectively. This signifies that AIV H5N1 used in this study is pathogenic and it is able to cause severe cytopathology to host cells even at low virus load.
Influenza, Human ; Influenza A Virus, H5N1 Subtype

Each year, influenza A infections have caused tremendous death rate as high as 300,000-500,000 globally. Althoughthere are effective anti-influenza agents and vaccines, high mutational rate among influenza A viruses renders dramaticdecline in the effectiveness of anti-influenza agents or vaccines in certain individuals. The situation is further complicatedby limitations in influenza vaccine production, for instance, long production period, limited vaccine capacity and lackof cross-protection against various influenza A virus strains. To solve these issues, development of universal influenzavaccine based on conserved antigens such as non-stuctural protein 1 (NS1) has been endeavoured. NS1 protein is highlyconserved in all influenza A virus strains known by far, produced abundantly on infected cell surfaces and responsible formaintaining virulence. Furthermore, cytotoxic T-lymphocytes that are active against NS1 were also reported to be ableto avoid shedding of influenza in hosts. To better inhibit influenza infections, oral immunization has long been proposeddue to feasibility of this method to be implemented and safer for recipients while able to target influenza A viruses fromthe entry point. Lactobacillus has been vastly studied for its roles as bacterial carrier in oral vaccine development dueto its significant probiotic properties. For examples, stimulation of immune responses in oral and airway mucosal layers,high colonization in oral and airway mucosal layers and great natural adjuvant effects. In this light, influenza universaloral vaccine developed using NS1 dan Lactobacillus should be further studied in influenza oral vaccine design.