Phenotypic and functional analysis of bovine peripheral blood dendritic cells before parturition by a novel purification method

Dendritic cells (DCs) are specialized antigen presenting cells specializing in antigen uptake and processing, and play an important role in the innate and adaptive immune response. A subset of bovine peripheral blood DCs was identified as CD172a⁺/CD11c⁺/MHC (major histocompatibility complex) class II⁺ cells. Although DCs are identified at 0.1%–0.7% of peripheral blood mononuclear cells (PBMC), the phenotype and function of DCs remain poorly understood with regard to maintaining tolerance during the pregnancy. All cattle used in this study were 1 month before parturition. We have established a novel method for the purification of DCs from PBMC using magnetic‐activated cell sorting, and purified the CD172a⁺/CD11c⁺ DCs, with high expression of MHC class II and CD40, at 84.8% purity. There were individual differences in the expressions of CD205 and co‐stimulatory molecules CD80 and CD86 on DCs. There were positive correlations between expression of cytokine and co‐stimulatory molecules in DCs, and the DCs maintained their immune tolerance, evidenced by their low expressions of the co‐stimulatory molecules and cytokine production. These results suggest that before parturition a half of DCs may be immature and tend to maintain tolerance based on the low cytokine production, and the other DCs with high co‐stimulatory molecules may already have the ability of modulating the T‐cell linage.     

A novel phenotype for Laron dwarfism in miniature Bos indicus cattle suggests that the expression of growth hormone receptor 1A in liver is required for normal growth

Mutations within the growth hormone receptor (GHR) gene that lead to an inactivated or truncated GHR protein cause abnormal growth and small adult size in a variety of species (Laron dwarfism). We studied a line of miniature Bos indicus cattle that have phenotypic (small mature size) and endocrine (increased blood growth hormone and decreased blood insulin-like growth factor-I concentrations) similarities to Laron dwarfs. Liver mRNA from miniature and control cattle was used to amplify a cDNA within the coding region of the GHR. The miniature cattle had GHR mRNA size (determined by Northern blot) and cDNA sequence that were similar to control cattle and, therefore, were unlike most Laron dwarf genotypes in which the GHR gene is mutated. Amounts of mRNA from liver as well as muscle (superficial neck and longissimus) were analyzed by ribonuclease protection assay for IGF-I, total GHR, GHR 1A (inducible, liver-specific GHR mRNA), and GHR 1B (constitutive GHR mRNA). Four control and five miniature bulls were tested. As expected, liver IGF-I mRNA was decreased in the miniature cattle (approximately 12% of control; P < 0.01). The amount of the total GHR as well as GHR 1A mRNA were also decreased in liver (17% and 19% of control, respectively; P < 0.01). Other GHR mRNA, including GHR 1B mRNA, were similar for miniature and control cattle. In muscle, there was a tendency (P < 0.10) for decreased IGF-I mRNA and increased GHR mRNA in miniature compared with control cattle. In summary, a novel phenotype for Laron dwarfism in Bos indicus cattle was associated with underexpression of GHR 1A mRNA, but not other GHR mRNA variants in liver. In addition to decreased GHR 1A mRNA, the miniature cattle had decreased liver IGF-I mRNA. Full expression of GHR 1A in liver, therefore, may be required for full liver IGF-I expression and normal growth.     

Supplementary report on experimental autumn crocus (Colchicum autumnale L.) poisoning in cattle: morphological evidence of apoptosis.

Previously we reported that tissue destruction characterized by the presence of karyopyknotic, karyorrhectic and mitotically arrested cells was seen in alimentary epithelial cells and lymphocytes in the lymphoid and hemopoietic systems of cattle experimentally administered with autumn crocus (Colchicum autumnale L.). This report deals with the mechanism of acute cellular injury following experimental autumn crocus poisoning in cattle as demonstrated by the in situ DNA strand break analysis and electron microscopy. The analyses revealed that cellular injury caused by autumn crocus was closely associated with apoptosis.     

Determination of maturity in male hawksbill turtle <Emphasis Type="Italic">Eretmochelys imbricata</Emphasis> in captivity based on tail elongation and plasma testosterone level

To determine the sexual maturity of fourteen male wild-caught hawksbill turtles [straight carapace length (SCL) range 63–79 cm] held in captivity, we investigated their hormone levels in blood and morphological characteristics. Male turtles were divided into two groups: five individuals showing mating behavior (group A) and nine individuals not showing mating behavior (group B). Then, seasonal changes of plasma testosterone of both groups were studied. We also tried to determine male maturity based on tail elongation (TE) index, i.e., the ratio of tail length (TL) to SCL. Plasma testosterone level of group A gradually increased during premating and then sharply declined to low levels between mating and postmating seasons. In contrast, that of group B remained low during the experimental periods. The distinct seasonal changes of plasma testosterone levels of groups A and B correlated with their maturity. Furthermore, males were clearly divided into mature and immature based on TE values >0.35 and <0.33, respectively, corresponding to results based on plasma testosterone levels. Our results indicate that TE can be used more conveniently and accurately to determine maturity of male hawksbill turtles in captivity.     

Germination and appressorium formation of Pyricularia oryzae Cavara can be inhibited by reduced concentration of Blasin®Flowable with carbon dioxide microbubbles

We investigated the possibility to reduce the usage of Blasin^®Flowable (BF), a disinfectant inhibiting the germination and appressorium formation of Pyricularia oryzae Cavara conidia, by using carbon dioxide microbubbles (CO₂MB). Germination was significantly inhibited by 10 000-fold diluted BF solution containing CO₂MB generated by the decompression-type generator compared to CO₂ millibubbles (CO₂MMB) and CO₂MB generated by the gas-water circulating-type generator. Appressorium formation in the 10 000-fold diluted BF solution containing both CO₂MBs was less than that in CO₂MMB. Scanning electron microscopy showed wrinkles and dents on the surface of conidia treated with 5 000-fold diluted BF solution containing both CO₂MBs. Via transmission electron microscopy, we observed the expansion of the vacuole and the intracellular space and bloated or absent lipid granules in the conidia treated with BF solution containing both CO₂MBs. Our results show that inhibition of the conidium germination and appressorium formation of P. oryzae Cavara by 10 000-fold diluted BF solution could be achieved by using the decompression-type CO₂MB.     

Assessing the acceptance of the system of rice intensification among farmers in rainfed lowland rice region of Cambodia

We conducted interview survey with rainfed rice farmers at a commune in southern Cambodia to identify the determinants of their adoption or rejection of the system of rice intensification (SRI) for the wet season of 2014. SRI was first introduced to the commune in 2003 and spread among the farmers, but the number of farmers discontinuing the practice has increased since 2011. We classified the farmers into four categories: those who practice SRI in all their fields (OA), farmers who practice SRI in some of their fields (PA), farmers who had practiced SRI but discontinued it (DA), and farmers who have never practiced SRI (CR). Farmers in different categories of SRI adoption differed in their access to water sources. The majority of OA and PA farmers had supplementary water sources, which was, however, available to less than 50% of DA farmers and only 15% of CR farmers. Both PA and DA farmers mentioned water shortage as the reason for not practicing SRI in some (PA) or any (DA) of their fields, but they differed in the number of rice fields. All the PA farmers had more than one field, whereas a majority of DA farmers had only one field. Labor shortage and difficulty of planting in a regular grid pattern were also mentioned as the reason for not practicing SRI by the farmers, particularly by DA farmers. Water constraint and difficulties in transplanting were thus identified as two major determinants of SRI adoption/rejection at the study site.