Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Effects of oral administration of colostrum whey in peripartum goat on antimicrobial peptides in postpartum milk

The present study was conducted to examine whether colostrum supplementation in peripartum goats increases the antimicrobial peptides in their milk. Goats were orally administered 2 ml of colostrum whey products (colostrum group) or water (control group) daily, from 2 weeks before until 2 weeks after kidding. Body weights of mothers and kids were measured. Blood, milk, and fecal samples were collected from the mothers, and blood samples were collected from the kids. Concentrations of milk antimicrobial peptides (beta‐defensin, cathelicidin, lactoferrin, S100A7, lactoperoxidase, and immunoglobulin A [IgA]) were determined. IgA and nutritional parameters (glucose, total cholesterol, triglyceride, ketone bodies, and non‐esterified fatty acids) were also determined in the blood of mothers and kids. Milk IgA and lactoferrin concentrations were higher in the colostrum group than in the control group. Conversely, lower milk concentrations of S100A7 were observed in the colostrum group than that in the control group. Plasma IgA concentrations were higher for kids from the colostrum group than for those from the control group. These results suggest that oral administration of colostrum in pregnant goats increases IgA concentration in postpartum milk, which can subsequently improve the health of their kids.     

Rediscovery of Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) MK.) an ancient crop of the Indian subcontinent

Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) Mac Key, synonym: T. sphaerococcum Perc.) is endemic to southern Pakistan and northwestern India. It was one of the main winter crops grown by ancient Indian cultures. However, it disappeared from the record during the early twentieth century, especially after the Green Revolution brought modern wheat varieties into India and Pakistan. Whether or not Indian dwarf wheat is presently cultivated has been unclear. Here we report on the rediscovery of the cultivation of this wheat in northern Karnataka and southern Maharashtra in India. Molecular genetic analysis of the chloroplast DNA of the two specimens collected at location 3 revealed that both samples have a unique haplotype that is specific to Indian dwarf wheat. We found this wheat at three locations in 2010, but at only one of the three locations in 2011. Therefore, the future survival of this subspecies is uncertain. Further ethnobotanical research is urgently needed to conserve this unique genetic resource for the future.     

Texture measurement of cabbages using an acoustical vibration method

Textures of six cabbage cultivars were quantified using an acoustical vibration technique. A sample of four outer leaves of a cabbage was penetrated using a probe. The acoustical vibration signals were measured during penetration using a piezoelectric sensor. A new texture index (TI), the “energy density”, was introduced, which was determined by the integration of squared amplitudes of texture signals multiplied by a factor of a frequency band. This TI enabled evaluation of acoustical signals in the high-frequency region (>1000 Hz) more sensitively than the previously used index (“amplitude density”), which was determined by the integration of texture signal amplitudes. Significant differences in TI among the cultivars were obtained by using ANOVA, especially between a spring and a winter cabbage. We also examined cabbages that had been stored under 4 °C for 10 or 19 d. Most TI readings increased after the storage. These results provide useful information related to the shelf-life of cabbages.     

Cytological characteristics of microconidia of <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

The inner cellular structure of microconidia of Magnaporthe oryzae was examined using fluorescent probes and electron microscopic techniques. The volume of the nucleus relative to the cell was significantly larger in microconidia than in macroconidia or vegetative hyphae, similar to observations for spermatia of other fungi. Selective fluorescent staining revealed that cytosolic RNA was less abundant in microconidia than in macroconidia and germ tubes, suggesting that general metabolic activity in microconidia is low. Consistently, GFP expression driven by the TrpC promoter was highly active during the formation of phialides and microconidia but gradually decreased as the microconidia matured. Such data suggest that microconidia are in a quiescent or dormant state.     

Proposal for stock-recruitment relationship for Japanese sardine <Emphasis Type="Italic">Sardinops melanostictus</Emphasis> in North-Western Pacific

ABSTRACT:   This study assessed the stock-recruitment relationship (SRR) for the Japanese sardine Sardinops melanostictus in the North-western Pacific. Of the 20 SRR models investigated, the Akaike information criterion (AIC) was the minimum (AIC = 551.2) when the data were separated into two groups (A and B) and the log-normal distribution was applied as the error term. Group A was constructed with SRR data from 1976–1987 and 1992–2004. Group B consisted of data from 1988–1991. The AIC minimum model was R  = 22.8 S  ×  e ^ε for Group A, where R , S , and ε denote the recruitment of sardine (individual number of 0-year old fish), spawning stock biomass (SSB), and error term, respectively. This model indicated that recruitment was proportional to the SSB and that no density-dependent effect operated over the range of SSB investigated (51 000–11.3 million t). Recruitment was markedly higher (lower) when the sea surface temperature (SST) of the Kuroshio Extension area in February was low (high). The essential SRR can simply be expressed as R  = 22.8 S  ×  e ^ε with the level of recruitment deviating from the model to a greater or lesser degree depending on the environmental conditions.     

Improvement in recruitment of Japanese sardine with delays of the spring phytoplankton bloom in the Sea of Japan

To evaluate the impact of temporal variation of primary productivity on the recruitment of Japanese sardine (Sardinops melanostictus) in the Sea of Japan, the phenology of sea surface phytoplankton abundance was estimated from 8 day multiple satellite (SeaWiFS, MODIS‐Aqua, MERIS, and VIIRS) derived sea surface chlorophyll (SSChl) a concentrations from January 1998 to December 2015. Because relationships between SSChl a and in situ chlorophyll a concentrations were significantly different among periods based on the satellite combinations used, maximum and minimum SSChl a concentrations of 1 year were relativized as 1 and 0, respectively. Spatio‐temporal variation of relativized SSChl a concentrations was determined by using empirical orthogonal function (EOF) analysis. Scores in the first EOF mode denoted the basin‐scale variations of SSChl a concentrations in the Sea of Japan, and the major peak from the end of February to the end of May displayed the spring bloom. The logarithm of recruitment per spawner (LNRPS) for sardine was positively affected by delays in the start and end dates of the spring phytoplankton bloom. The delay of the date of the lowest sea surface temperature contributed to the delay of the end of the spring bloom during the period 1998–2015 and elevated the LNRPS during the period 1982–2015. Sardine spawns in the southern Sea of Japan from April to May, hence, delays of the spring bloom prolonged its overlap with sardine larval periods, and thus improved the recruitment of Japanese sardine in the Sea of Japan.     

Serum vascular endothelial growth factor in dogs with various proliferative diseases

Angiogenesis plays an important role in the proliferation and metastasis mechanisms of malignant tumors. Vascular endothelial growth factor (VEGF), a group of cytokines that contribute to angiogenesis and vasculogenesis. This study aimed to investigate the serum VEGF-A concentrations in dogs with various proliferative diseases. A total of 202 dogs that were histopathologically diagnosed with proliferative diseases were included in the study. Serum VEGF-A concentrations were measured using enzyme-linked immunosorbent assay. Median serum VEGF-A concentrations in dogs were as follows: healthy dogs, 4 pg/ml [0–21 pg/ml]; hepatocellular carcinoma, 30 pg/ml [0–158 pg/ml, P=<0.001]; hepatocellular adenoma, 32 pg/ml [0–49 pg/ml, P=0.003]; hepatic nodular hyperplasia, 18 pg/ml [0–51 pg/ml, P=0.595]; adrenal pheochromocytoma, 32 pg/ml [0–187 pg/ml, P=<0.001]; adrenocortical carcinoma, 32 pg/ml [3–161 pg/ml, P=0.002]; adrenocortical adenoma, 27 pg/ml [0–106 pg/ml, P=0.005]; colorectal adenocarcinoma, 36 pg/ml [0–75 pg/ml, P=0.002]; colorectal adenoma, 43 pg/ml [0–48 pg/ml, P=0.144]; inflammatory colorectal polyps, 37 pg/ml [0–111 pg/ml, P=<0.001]; pulmonary adenocarcinoma, 35 pg/ml [4–107 pg/ml, P=0.002]; pulmonary histiocytic sarcoma, 35 pg/ml [0–131 pg/ml, P=0.016]; and follicular thyroid carcinoma, 35 pg/ml [0–106 pg/ml, P=0.009]. The serum VEGF-A concentrations were significantly higher in dogs with neoplastic lesions compared to healthy dogs, except for colorectal adenoma. High serum VEGF-A concentrations were observed in dogs with proliferative diseases. The present study suggests that angiogenesis-inhibiting therapy, which targets VEGF-A, may be useful for canine neoplastic diseases.     

Hematology in sika deer (Cervus nippon yesoensis Heude, 1884)

Blood samples were taken from 78 wild and 21 farmed sika deer (Cervus nippon yesoensis) using ketamine-xylazine sedation during their excited (82 deer) and resting (17 deer) states. Red cell count (RBC), hemoglobin (Hb), packed cell volume (PCV) and mean corpuscular volume (MCV) were significantly higher and mean corpuscular hemoglobin concentration (MCHC) was lower in excited deer than in resting deer. There was no significant difference in total leukocyte count (WBC) between excited and resting wild males, while a marked increase of WBC with neutrophilia was observed in excited wild females. RBC and PCV were significantly higher and MCH was lower in excited males than in excited females. In wild deer, WBC was significantly higher in females than in males, but there was no significant difference in WBC between farmed males and females. Sex differences in the hematological parameters were not observed in fawns (10 months).