Evaluation of the PATHFAST Chemiluminescent Enzyme Immunoassay for Measuring Progesterone in Whole Blood and Serum of Mares

Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone in whole blood and serum samples were measured by PATHFAST. In addition, concentrations of progesterone in serum samples measured by PATHFAST were compared with those measured by radioimmunoassay (RIA) and enzyme immunoassay (EIA). Using PATHFAST, the serum concentrations of progesterone in mares correlated highly with those of whole blood samples (r=0.9672, n=88). The serum concentrations of progesterone as measured by PATHFAST correlated well with RIA (r=0.9654, n=88) and EIA (r=0.9323, n=112). An abrupt decline in circulating progesterone in whole blood samples was observed within 2 hr (50%), followed by a gradual decline until 48 hr later. The results for progesterone in whole blood samples correlated highly with those in serum samples, and the declining pattern paralleled that of the serum samples. These results demonstrated that PATHFAST is useful in the equine clinic as an accurate diagnostic tool for rapid assay of progesterone within 26 min, using unextracted whole blood.     

Phenotypic changes in reproductive traits with changes in stock size of the starspotted smooth-hound Mustelus manazo in Tokyo Bay, Japan

Reproductive traits are among the primary determinants of population dynamics, and their changes could affect stock size. We investigated differences in reproductive traits of the starspotted smooth-hound Mustelus manazo in Tokyo Bay between the 1990s (a low-stock period) and the 2000s (a high-stock period). Peak timings of mating (May–July), ovulation (May–June), and parturition (May–June) were similar between the two periods. However, the size at first maturity in the 2000s was smaller than that in the 1990s for both sexes. In addition, embryo-related parameters differed greatly between the two periods. Slopes of size–fecundity regression in the 2000s decreased to 43–73 % of the 1990–1996 values, suggesting a substantial decline in fecundity during the 2000s. Frequency distribution of the maximum ovum diameter during the ovulation season showed two distinct two modes in 1990s while it showed a substantial dispersion during the 2000s, implying that changes occurred in the developmental pattern of ova in the ovary. Undeveloped eggs were present in the uteri with low frequency from September to May during the 2000s, but they were totally absent from September through May in the 1990s. The increased number of remnant undeveloped eggs that did not become embryos may explain the fecundity decline during the 2000s.     

Yield-enhancing and tuber-downsizing effects of transplantation cultivation method of case-held tuber seedlings in the sweet potato cultivar Beniharuka

We developed transplantation cultivation method of case-held tuber seedlings (CTS), which was derived from direct planting method of seed tubers, and applied this method to the sweet potato cultivar Beniharuka. A plastic case made of polypropylene was designed for cultivation of CTS. Seed tubers of cultivar Beniharuka in the range of 30–80 g were cut in half. The half-cut tubers were placed inside the plastic cases, and the cases were filled with a commercial soil mix. The case-held tubers were incubated under natural sunlight in a glass house. After 3–4 wk, the CTS were transplanted into a field. Mother tuber (seed tuber) enlargement was suppressed by the plastic confinement of the cases, and daughter tubers were formed above the case as vine-root-originated tubers. In the field experiments in 2012 and 2013, daughter tuber yields were increased 19% and 21% by case-held tuber seedling transplanting (CTST) over conventional vine-planting (VP), the number of daughter tubers per plant in CTST were 36 and 68% higher than in VP, and the mother tuber yields were limited to 2.1 and 4.3% of the total fresh yield of mother and daughter tubers, respectively in 2012 and 2013. Application of CTST method to cultivar Beniharuka enhanced tuber yield, increased the number of daughter tubers per plant, downsized daughter tubers compared to VP, and mother tuber enlargement was suppressed by case-holding. The CTST method is expected to produce more and smaller good in shape tubers of cultivar Beniharuka compared to VP.     

Theileria-free grazing of dairy heifers on grassland in Kyushu,Japan where T. orientalis was epidemic before a 7-year vacancy

This study aimed to produce a Theileria-free grazing system for Holstein heifers reared on a dairy farm in the Hita area, Kyushu, Japan. In the grazing area, spreading of T. orientalis infection was confirmed in 2009. To eradicate the T. orientalis infection, four measures were conducted: 1) 7-year deferred grazing; 2) grazing only T. orientalis-uninfected heifers; 3) anemia check by red blood cell parameters at least once per month; and 4) protecting heifers from blood-sucking T. orientalis-infected ticks. Grazing was restarted in 2017 in the same area and continued to 2021. During last 2 years of pasturing (2020–2021) all of the 129 heifers were confirmed to be T. orientalis-free. In summary, it is possible to establish a T. orientalis-free grazing system by conducting appropriate measures.     

Rainfall,Stemflow, and Throughfall Chemistry at Urban- and Mountain-facing Sites at Mt. Gokurakuji,Hiroshima, Western Japan

Rainfall, stemflow, and throughfall were collected from 1996 to 1999 at two types of forest sites: (1) forests near the traffic roads and urban areas and (2) forests away from the urban areas at Mt. Gokurakuji, Hiroshima, western Japan in order to estimatethe effects of anthropogenic activities on atmospheric deposition. Rainfall deposition for major ions showed small differences between the sites. The NO₃ ^- and SO₄ ^2-concentrations in stemflow were higher at the urban-facing slope than at the mountain-facing slope. Throughfall deposition of NO₃ ^- and SO₄ ^2- was also higher at urban-facing slopes. Net throughfall (NTF) deposition (throughfall minus rainfall) of NO₃ ^- and SO₄ ^2- accounted for 77 and50% of the total throughfall deposition on urban-facing slopes, respectively, while it accounted for 44 and 23% on themountain-facing slopes, respectively. These results indicated a higher contribution from dry deposition on urban-facing slopes compared to mountain-facing slopes. Atmospheric N (NO₃ ^- +NH₄ ⁺) deposition from throughfall was estimated to be around 17–26 kg N ha^-1 yr^-1 on urban-facing slopes, which was greater than the threshold of N deposition that could cause nitrogen leaching in Europe and the United States. The highload of atmospheric N deposition may be one of the factors bringing about the decline of pine forests on urban-facing slopesof Mt. Gokurakuji.     

Effect of water activity on the release characteristics and oxidative stability of D-limonene encapsulated by spray drying

The stability of encapsulated D-limonene, which was prepared by spray drying, was studied in view of the release characteristics and oxidation stability. Gum arabic, soybean water-soluble polysaccharide, or modified starch, blended with maltodextrin were used as the wall materials. The powders were stored under the conditions of 23-96% relative humidity at 50 degrees C. The release rate and the oxidation rate were closely related to the relative humidity. The relationship was not simple. Initially, the release rate and the oxidation rate increased with increasing water activity, but around the glass transition temperature, the rates decreased sharply to increase again at a further increase of water activity. The results could be explained by a change in the powder structure, where a glass capsule matrix was changed into rubbery state during storage.     

Thermal resistance traits of the clonal silver crucian carp Carassius langsdorfii and evaluation of these traits using primary culture cells

The thermal resistance traits of three clonal lines of the silver crucian carp Carassius langsdorfii (SCC-1, -2 and -3) were investigated. Individual juvenile fish reared at 20°C were exposed to thermal stress at 36.5 ± 0.5°C and their death times during this exposure were measured. The death times of SCC-2 and SCC-3 fish were 66.6 ± 31.2 and 144.7 ± 49.8 min, respectively. In contrast, all SCC-1 fish survived under these conditions. Furthermore, the thermal sensitivity of primary culture cells from each clone was determined at 37, 40 or 43°C by the Trypan blue assay. Under all treatment conditions, the thermal sensitivity of the SCC-1 primary cultures was lower than those of the others. These results estimated the correlation between the in vivo and in vitro thermal resistances. Therefore, the use of primary culture cells to evaluate thermal resistance could be a useful method of selection breeding.