Tolerance and safety of Lactobacillus paracasei ssp. paracasei in combination with Bifidobacterium animalis ssp. lactis in a prebiotic-containing infant formula: a randomised controlled trial.

The addition of probiotics to infant formula has been shown to be an efficient way to increase the number of beneficial bacteria in the intestine in order to promote a gut flora resembling that of breast-fed infants. The objective of the present study was to evaluate the safety and tolerance of a combination of two probiotic strains in early infancy. A group of 126 newborns were randomised to receive a prebiotic-containing starter formula supplemented with Lactobacillus paracasei ssp. paracasei and Bifidobacterium animalis ssp. lactis or the same formula without probiotics for the first 3 months of life. A total of eighty infants continued the study until they were aged 6 months. Growth measurements were taken monthly at healthy baby clinics. Diaries were used to monitor behaviour, infections, use of antibiotics, as well as stool characteristics. Normal growth occurred in all infants and no statistically significant differences were detected between the probiotics group and the control group for gain in weight, length and head circumference. Infants in the probiotics group produced softer and more frequent stools during the first 3 months of life. No differences were found in crying and sleeping hours, number of parent-diagnosed infections, antibiotic use, visits to the general practitioner and number of adverse events. The use of a prebiotic-containing starter formula supplemented with L. paracasei ssp. paracasei and B. animalis ssp. lactis in early infancy is safe, well tolerated and has no adverse effects on growth and infant behaviour.

Phylogeny of fungal hemoglobins and expression analysis of the Aspergillus oryzae flavohemoglobin gene fhbA during hyphal growth.

The fhbA genes encoding putative flavohemoglobins (FHb) from Aspergillus niger and Aspergillus oryzae were isolated. Comparison of the deduced amino acid sequence of the A. niger fhbA gene and other putative filamentous fungal FHb-encoding genes to that of Ralstonia eutropha shows an overall conserved gene structure and completely conserved catalytic amino acids. Several yeasts and filamentous fungi, including both Aspergillus species have been found to contain a small FHb gene family mostly consisting of two family members. Based on these sequences the evolutionary history of the fungal FHb family was reconstructed. The isolated fhbA genes from A. oryzae and A. niger belong to a phylogenetic group, which exclusively contains Aspergillus genes. Different experimental approaches show that fhbA transcript levels appear during active hyphal growth. Moreover, in a pclA-disrupted strain with a hyperbranching growth phenotype, the transcript levels of the fhbA gene were 2­5 times higher compared to the wild-type. These results suggest that FHb from filamentous fungi have a function that is correlated to the hyphal growth phenotype.

Impact of milk consumption on performance and health of primary school children in rural Vietnam.

This is a follow-up study to an investigation on the prevalence of malnutrition and micronutrient deficiencies among Vietnamese primary schoolchildren. A total of 454 children aged 7 to 8 years attending three primary schools in the Northern delta province of Vietnam were either provided with regular milk, milk fortified with vitamins, minerals and inulin or served as a reference control group. Children were monitored for anthropometrics, (micro)-nutritional status, faecal microbiota composition, school performance, and health indices. Both weight-for-age (WAZ) and height-for-age (HAZ) significantly improved during 6 months of milk intervention; and underweight and stunting dropped by 10% in these groups. During intervention the incidence of anemia decreased and serum ferritin levels increased significantly in all groups. Serum zinc levels increased and consequently the incidence of zinc deficiency improved significantly in all three groups. Serum retinol levels and urine iodine levels remained stable upon intervention with fortified milk whereas in the control group the incidence of iodine deficiency increased. Bifidobacteria composed less than 1% of the total faecal bacteria. After three months of milk intervention total bacteria, bifidobacteria and Bacteroides sp. increased significantly in both milk and inulin fortified milk groups. Children in the milk consuming groups had significantly better short-term memory scores. Parent reported that health related quality of life status significantly improved upon milk intervention. In conclusion, (fortified) milk consumption benefited the children in rural Vietnam including lowering the occurrence of underweight and stunting, improving micronutrients status and better learning indicators as well as improving the quality of life.

Expression of Aspergillus hemoglobin domain activities in Aspergillus oryzae grown on solid substrates improves growth rate and enzyme production.

DNA fragments coding for hemoglobin domains (HBD) were isolated from Aspergillus oryzae and Aspergillus niger. The HBD activities were expressed in A. oryzae by introduction of HBD gene fragments under the control of the promoter of the constitutively expressed gpdA gene. In the transformants, oxygen uptake was significantly higher, and during growth on solid substrates the developed biomass was at least 1.3 times higher than that of the untransformed wild-type strain. Growth rate of the HBD-activity-producing strains was also significantly higher compared to the wild type. During growth on solid cereal substrates, the amylase and protease activities in the extracts of the HBD-activity-producing strains were 30-150% higher and glucoamylase activities were at least 9 times higher compared to the wild-type strain. These results suggest that the Aspergillus HBD-encoding gene can be used in a self-cloning strategy to improve biomass yield and protein production of Aspergillus species.

Aspergillus oryzae in solid-state and submerged fermentations. Progress report on a multi-disciplinary project.

We report the progress of a multi-disciplinary research project on solid-state fermentation (SSF) of the filamentous fungus Aspergillus oryzae. The molecular and physiological aspects of the fungus in submerged fermentation (SmF) and SSF are compared and we observe a number of differences correlated with the different growth conditions. First, the aerial hyphae which occur only in SSFs are mainly responsible for oxygen uptake. Second, SSF is characterised by gradients in temperature, water activity and nutrient concentration, and inside the hyphae different polyols are accumulating. Third, pelleted growth in SmF and mycelial growth in SSF show different gene expression and protein secretion patterns. With this approach we aim to expand our knowledge of mechanisms of fungal growth on solid substrates and to exploit the biotechnological applications.