Development of molecular immunoassay system for probiotics via toll-like receptors based on food immunology

Recent interest has focused on the importance of intestinal immunity for the host defense, but to date, not much is known about the underlying mechanisms. The toll‐like receptor (TLR) family plays an important role in host defense through recognizing bacterial pathogen‐associated molecular patterns. Our recent research on the physiological function of food products has investigated the immunoregulatory effects of probiotic lactic acid bacteria (LAB) via TLR. Studies of swine, which often substitute for a human model, have demonstrated intestinal immunoregulation by the probiotic LAB mediated by TLR in the gut. On the basis of our study, efforts have also been made to develop a molecular immunoassay system for probiotic LAB and find novel immunostimulatory DNA sequences from probiotics and high potential immunobiotic LAB strains via TLR signaling. These findings may provide important clues at the molecular level on TLR signal transduction pathways and recognition mechanisms for the ligands. They also provide impetus to further delineate the activation mechanism of the innate immune response. In addition to identifying immunoregulatory factor immunogenics from LAB, a better understanding of intestinal immune regulation through cytokine networks holds out promise for basic food immunology research and the development of immunobiotic foods to prevent specific diseases.     

Nucleotide sequence polymorphisms of beta1‐, beta2‐, and beta3‐adrenergic receptor genes on Jinhua,Meishan, Duroc and Landrace pigs

The full amino acid coding sequences of adrenergic receptor genes beta1, beta2, and beta3 (ADRB1, ADRB2, and ADRB3)were determined for Jinhua, Meishan, Duroc and Landrace pigs. Non‐synonymous substitution of Arg458Pro was found in the porcine ADRB1 gene, resulting in a 469 amino acid sequence. Continuous substitutions of Asn29Asp and Glu30Gln were found in the porcine ADRB2 gene, resulting in a 418 amino acid sequence. Additionally, a Lys30 polymorphism of the ADRB2 gene was found in the Jinhua pigs. There were three non‐synonymous substitutions of Asn24Thr, Arg264Gln and Asn398Asp on the porcine ADRB3 gene. A thymine insertion in the ADRB3 gene, resulting in a protein with two fewer amino acids, was found in the Jinhua and Meishan pigs. To assess the effect of ADRB polymorphisms on porcine subcutaneous fat layer thickness, we calculated the genetic frequency of the variants in fatty and lean groups, each consisting of 24 pigs that were crossbreds of Duroc and Jinhua pigs. The effect of the ADRB3 gene polymorphism was not evaluated, because there was insufficient variation on the ADRB3 gene in the examined groups. Although Fisher's exact test showed no significant difference in the frequency of ADRB1 and ADBR2 variants between the two groups, the Arg458 variant of ADRB1 was higher (P = 0.11) in the lean group, and pigs in that group had a thinner fat layer than did those with the Pro458 variant. These results imply a possibility of ADRB1 polymorphism as a minor factor in porcine fat layer thickness. The Asp29 variant of ADRB2 was higher in the lean group (P = 0.11), and the Glu30 variant was higher in the fatty group (P = 0.15), but the Asp29 variant was found only in the Chinese pigs. Thus, the effect of ADRB2 polymorphisms was not clear in this study.     

Monoecy and gynomonoecy in Spinacia oleracea L.: Morphological and genetic analyses

Two cosexual spinach lines, 03-336 and 03-258, were examined to investigate their sex expression. We found that line 03-336 consisted of plants bearing both pistillate and staminate flowers on the same individual (strictly monoecious type), while line 03-258 consisted of plants with both pistillate and hermaphroditic flowers on the same individual (gynomonoecious type). Interestingly, the two lines exhibited distinct patterns of inheritance when crossed with the pistillate plants derived from a dioecious line. The strictly monoecious character was considered to be conditioned by the known allele X^m, whereas the gynomonoecious character appeared to be governed by a novel monoecious allele or modifying gene(s) that may affect the sex expression.     

Intranasal tumor of the ethmoid olfactory mucosa in sheep.

Intranasal tumors (papillary adenomas or adenocarcinomas) of the ethmoid olfactory mucosa of sheep were investigated by light and electron microscopy. The fine structure of the tumor cells was characterized by the presence of numerous secretory granules. Viral particles, which were morphologically similar to a visna-maedi virus, were detected in all tumor tissues and in 3 of 4 cultures examined. The particles (about 97 nm) had an eccentrically located electron-dense core and numerous spikes on their surfaces. The RNA-dependent DNA polymerase activities in the tumor cells or the cultured cell from the tumor were greater than those in the normal intranasal tissues or the cultured cells from the choroid plexus. Viral particles similar to herpesvirus were also detected in 1 culture.     

Effects of heat stress on production,somatic cell score and conception rate in Holsteins

We examined the effects of heat stress (HS) on production traits, somatic cell score (SCS) and conception rate at first insemination (CR) in Holsteins in Japan. We used a total of 228 242 records of milk, fat and protein yields, and SCS for the first three lactations, as well as of CR in heifers and in first‐ and second‐lactation cows that had calved for the first time between 2000 and 2012. Records from 47 prefectural weather stations throughout Japan were used to calculate the temperature–humidity index (THI); areas were categorized into three regional groups: no HS (THI < 72), mild HS (72 ≤ THI < 79), and moderate HS (THI ≥ 79). Trait records from the three HS‐region groups were treated as three different traits and trivariate animal models were used. The genetic correlations between milk yields from different HS groups were very high (0.91 to 0.99). Summer calving caused the greatest increase in SCS, and in the first and second lactations this increase became greater as THI increased. In cows, CR was affected by the interaction between HS group and insemination month: with summer and early autumn insemination, there was a reduction in CR, and it was much larger in the mild‐ and moderate‐HS groups than in the no‐HS group.