Determination of nucleotides in infant formula by ion-exchange liquid chromatography

Nucleotide-supplemented infant formula has been shown to positively modify the composition of intestinal microflora, emulating the attribute of human milk. Quantification of nucleotides in infant formula is of interest because of its applicability in quality and safety assessments. There is no standard method for the analysis of nucleotides in infant formula. In the present study, ion-exchange liquid chromatography (IELC)- and centrifugal ultrafiltration (CUF)-based protocols were developed for routine determination of additive nucleotides in infant formula. Five target nucleotides, guanosine 5'-monophosphate (GMP), inosine 5'-monophosphate (IMP), uridine 5'-monophosphate (UMP), cytidine 5'-monophosphate (CMP), and adenosine 5'-monophosphate (AMP) were measured by IELC with a mobile phase of 50 mM diammonium hydrogen phosphate buffer, pH 4.0, with UV detection at 254 nm. The calibration was linear over the range 0.5-50 microg/mL; R(2) = 0.999. The calculated LOD and LOQ were 0.01-0.05 microg/mL and 0.05-0.5 microg/mL, respectively. Recovery values (spiked concentration levels: 0.5, 5, and 10 microg/mL) ranged from 85.0 +/- 1.4% to 92.3 +/- 2.1% using only CUF preparation. This was applied to measure the concentration of five nucleotides in common infant formulas.     

A radio transmission pH measurement system for continuous evaluation of fluid pH in the rumen of cows

We developed a novel wireless radio transmission pH measurement system to continuously monitor ruminal bottom pH in cows, and compared these measurements to pH values determined by a spot-sample method. The wireless system consists of a pH sensor, data measurement receiver, relay unit, and personal computer with special software. The bullet-shaped sensor can be easily administered orally via a catheter into the rumen, without surgery. The glass electrode, using a temperature compensation system, can detect the rumen fluid pH with high accuracy. The ruminal bottom pH in healthy rumen-fistulated cows was measured as 6.52 ± 0.18 by the wireless system and as 6.62 ± 0.20 by the spot-sample method; with a correlation between pH measurements using these different methods (n = 8, 24 samples, r = 0.952, P < 0.01). When measured serially in a cow fed a diet evoking rumen acidosis, the ruminal bottom pH decreased markedly following the morning feeding and then increased gradually by the next morning feeding. This wireless system is a ready-to-use tool for estimating circadian changes in ruminal bottom pH.     

3-Methylthiopropionic acid ethyl ester, isolated from Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon), enhanced differentiation in human colon cancer cells

The fully ripened fruit of Katsura-uri Japanese pickling melon ( Cucumis melo var. conomon) has rarely been used for food because the midripened fruit is utilized for making pickles, but the fully ripened fruit is no longer valuable for pickles due to the fruit body being too soft. We have considered the utilization of the fully ripened Katsura-uri fruit that may be used for nonpickling products, particularly if the fully ripened fruit demonstrated health benefits such as anticarcinogenic properties. The phytochemical extract from the fully ripened fruit of Katsura-uri Japanese pickling melon was purified via a bioassay-guided fractionation scheme, which was based on the induction of differentiation in a RCM-1 human colon cancer cell line. On the criteria of two differentiation markers (duct formation and alkaline phosphatase activity), the most potent fraction contained a compound identified as 3-methylthiopropionic acid ethyl ester, based on GC retention time, EI-MS, (1)H NMR, and (13)C NMR spectra. Previously, the role of 3-methylthiopropionic acid ethyl ester was considered as an odor producing compound in many fruits, but this study indicates potential medical benefits of this compound.     

Comparative studies of the quantification of genetically modified organisms in foods processed from maize and soy using trial producing

Seven types of processed foods, namely, cornstarch, cornmeal, corn puffs, corn chips, tofu, soy milk, and boiled beans, were trial produced from 1 and 5% (w/w) genetically modified (GM) mixed raw materials. In this report, insect resistant maize (MON810) and herbicide tolerant soy (Roundup Ready soy, 40-3-2) were used as representatives of GM maize and soy, respectively. Deoxyribonucleic acid (DNA) was extracted from the raw materials and the trial-produced processed food using two types of methods, i.e., the silica membrane method and the anion exchange method. The GM% values of these samples were quantified, and the significant differences between the raw materials and the trial-produced processed foods were statistically confirmed. There were some significant differences in the comparisons of all processed foods. However, our quantitative methods could be applied as a screening assay to tofu and soy milk because the differences in GM% between the trial-produced processed foods and their raw materials were lower than 13 and 23%, respectively. In addition, when quantitating with two primer pairs (SSIIb 3, 114 bp; SSIIb 4, 83 bp for maize and Le1n02, 118 bp; Le1n03, 89 bp for soy), which were targeted within the same taxon specific DNA sequence with different amplicon sizes, the ratios of the copy numbers of the two primer pairs (SSIIb 3/4 and Le1n02/03) decreased with time in a heat-treated processing model using an autoclave. In this report, we suggest that the degradation level of DNA in processed foods could be estimated from these ratios, and the probability of GM quantification could be experimentally predicted from the results of the trial producing.     

Economic loss due to treatment of bovine respiratory disease in Japanese Black calves arriving at a backgrounding operation in Miyazaki

The present study assessed the economic loss due to treatment of bovine respiratory disease (BRD) in Japanese Black calves that were introduced to a backgrounding operation from the age of 3 to 4 months until the age of 10 months. The data collected from a backgrounding operation in Miyazaki, Japan included the records of 2,690 animals entering the farm from 2013 to 2018. The treatment duration was defined as the number of days from the beginning to end of treatment. The cost of treatment was defined as the total cost of treatment during the treatment duration. The average incidence of BRD was 54.6%, and the relative frequency of calves that had BRD once, twice, and three or more times until they left the farm was 64.4%, 26.8%, and 8.8%, respectively (total recorded diagnoses of BRD: 2,494). Among the 2,494 recorded diagnoses, the average and median duration of treatment of BRD was 5.9 days and 3.0 days, respectively. The average and median cost of treatment was 7,767 and 5,600 Japanese yen, respectively. A prolonged duration of treatment and high cost of treatment were associated with BRD relapse, steers, and early stage of production (P<0.05). At the studied farm, the total cost of treatment during the 6-year study period was 19,658,988 yen, and the annual cost was approximately 3 million yen. In summary, the present study showed that BRD had a large economic impact in this backgrounding operation.     

Daily age and food habits of Lateolabrax latus larvae and juveniles occurring in the innermost shallow waters of Uwajima Bay, Japan

ABSTRACT:   Daily age and food habits of Lateolabrax latus larvae and juveniles were investigated at two different habitats in Uwajima Bay, western Shikoku, Japan. In the shallow zone at Miura, which contained eelgrass beds, larvae occurred from January and were collected abundantly in March. The size, age, and hatch-date distributions suggested that late-hatched specimens stay longer than early-hatched specimens at this station. On the contrary, in the intertidal flats of the Kunomura River mouth larvae were rarely collected, but juveniles were collected in relative abundance from April. The distributions in the intertidal flats station suggested that specimens had migrated from another location and then resided in the intertidal flats. Spatial and temporal trophic variations increased remarkably with metamorphosis, and thus, the spatial differences in growth rate were considered in relation to the trophic diversity of prey items. Our results suggest that the nursery environment, nursery utilization pattern, and food habits of early life stages of L. latus show remarkable flexibility. From the similarity of these ecological aspects with L. japonicus , which is a closely related species, we suggest the important role of the interspecific competition to the realized niche difference between Lateolabrax congeners.     

3-(4-Hydroxyphenyl)propionic acid is involved in the biosynthesis of myricanol in Myrica rubra

There is little evidence concerning the biosynthetic pathways for cyclic diarylheptanoids. We previously demonstrated that the cyclic diarylheptanoids myricanol and myricanone were biologically synthesized from two molecules of 4-coumaric acid by the feeding of 4-[8,9-¹³C₂]coumaric acid to young shoots of Myrica rubra. In the present study, using a ¹³C-labeled compound, we revealed that two molecules of 3-(4-hydroxyphenyl)propionic acid could also be a biosynthetic precursor of myricanol in M. rubra. These results indicated that both 4-coumaric acid and its dihydro-derivative were incorporated into myricanol. Competitive feeding experiments with 4-[8,9-¹³C₂]coumaric acid and 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid were performed in M. rubra to determine the preferential incorporation of these two precursors. ¹³C-NMR studies indicated that 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid was preferentially incorporated into myricanol. The data provided evidence for a biosynthetic sequence originating from 4-coumaric acid and leading to myricanol, through 3-(4-hydroxyphenyl)propionic acid, in M. rubra.