Reference values of hematological and biochemical parameters for the world smallest microminipigs

In this study, we demonstrated growth curves and reference values for hematological and serum biochemical parameters of Microminipigs, the world smallest experimental minipigs. In both male and female animals, the body weights (BWs) at 3 and 6 months of age were <5 kg and <10 kg, respectively, and growth curve revealed almost plateau (approximately 20 kg BW) after 18 months of age. Major hematological and serum biochemical parameters showed no gender differences and the values were very similar to those in G?ttingen and Yukatan minipigs. The values obtained in this study can serve as fundamental reference, and thereby facilitate the use of Microminipig in life science research.     

Optimization of oxygen concentration for growing bovine oocytes in vitro: constant low and high oxygen concentrations compromise the yield of fully grown oocytes

The oxygen environment in cell culture has a significant impact on the health and performance of cells. Here, we compared the effects of reduced (5%) and ambient (20%) oxygen concentrations on bovine oocyte-granulosa cell complexes, each containing a growing oocyte 90-102 μm in diameter, cultured for 14 days. Both oxygen concentrations showed some advantages and disadvantages; in 5% oxygen, the survival rate of oocytes was significantly higher than in 20% oxygen, but the resulting oocytes were significantly smaller, which was a serious disadvantage. During the first 4 days of culture, the growth and viability of oocytes were satisfactory using 5% oxygen. This observation led us to examine the effect of changing the oxygen concentration from 5% to 20% on Day 4 in order to minimize the expected disadvantages of constant 5% and 20% oxygen. The largest population of fully grown oocytes was obtained from cultures in which the oxygen concentration was changed in this way, which also led to higher oocyte viability than in constant 20% oxygen. A similar tendency was found in the frequency of oocytes becoming blastocysts after in vitro fertilization. Surviving oocytes eventually became located within an enlarged dome-like structure, and although the 5% oxygen environment may have been appropriate for oocyte growth in the early stages, 20% oxygen may have been necessary for the growth of oocytes in the dome-like structure. These results indicate an effective way of modulating oxygen concentration according to the growth of oocyte-granulosa cell complexes in vitro.     

Patterns of root respiration rates and morphological traits in 13 tree species in a tropical forest

The root systems of forest trees are composed of different diameters and heterogeneous physiological traits. However, the pattern of root respiration rates from finer and coarser roots across various tropical species remains unknown. To clarify how respiration is related to the morphological traits of roots, we evaluated specific root respiration and its relationships to mean root diameter (D) of various diameter and root tissue density (RTD; root mass per unit root volume; gcm(-3)) and specific root length (SRL; root length per unit root mass; mg(-1)) of the fine roots among and within 14 trees of 13 species from a primary tropical rainforest in the Pasoh Forest Reserve in Peninsular Malaysia. Coarse root (2-269mm) respiration rates increased with decreasing D, resulting in significant relationships between root respiration and diameter across species. A model based on a radial gradient of respiration rates of coarse roots simulated the exponential decrease in respiration with diameter. The respiration rate of fine roots (<2mm) was much higher and more variable than those of larger diameter roots. For fine roots, the mean respiration rates for each species increased with decreasing D. The respiration rates of fine roots declined markedly with increasing RTD and increased with increasing SRL, which explained a significant portion of the variation in the respiration among the 14 trees from 13 species examined. Our results indicate that coarse root respiration in tree species follows a basic relationship with D across species and that most of the variation in fine root respiration among species is explained by D, RTD and SRL. We found that the relationship between root respiration and morphological traits provides a quantitative basis for separating fine roots from coarse roots and that the pattern holds across different species.     

Growth of bovine oocyte-granulosa cell complexes cultured individually in microdrops of various sizes

In mammalian embryo culture, the embryo:medium volume ratio can substantially affect embryo developmental performance. In the present study, we tested the possibility of improving the growth of bovine oocytes by reducing the medium volume, from a typical volume used in mouse follicle culture to a minimum possible level. A total of 282 complexes, each containing a growing oocyte 87-100 mum in diameter, were individually placed in microdrops of 2, 5, 10 or 20 microl and cultured for 13 days in a modified TCM-199 supplemented with 4% polyvinylpyrrolidone (molecular weight: 360 kDa). Oocyte diameter was measured every other day to trace the growth of each oocyte. Half the medium was replaced every other day or every day, and comparison revealed that daily replacement was more favorable for culture of these microdrops. The highest survival rate, 95%, occurred in the 20-microl microdrops, where most oocytes continued to grow throughout the culture period. In comparison, in the 5- and 10-microl microdrops, more oocytes died, and growth slowed towards the end of culture. In the 2-microl microdrops, which had the highest death rate, growth virtually ceased after 9 days. The surviving oocytes were usually accompanied by a characteristic dome-like structure of the granulosa cell mass, except in the 2-microl microdrops. In conclusion, the 20-microl microdrops allowed oocyte growth at an acceptable level, and any further reduction of the volume only had a negative impact on oocytes.     

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Lingual antimicrobial peptide (LAP), a member of the β-defensin family in cows, is involved in the innate immune system and plays a crucial role in killing a large variety of microorganisms. The aim of the present study was to demonstrate the immunolocalization of LAP in the mammary glands of cows. A LAP antibody was raised in a rabbit by immunity with a synthetic 11 amino acid sequence out of a 42-amino acid sequence of the mature form of LAP. The specificity of the LAP antibody was checked using a competitive immunoassay and Western blotting. Paraffin sections of the mammary gland were immunostained with LAP antibody. In the competitive immunoassay, an increase of synthetic LAP concentration suppressed the optical density. Western blotting analysis for LAP revealed the presence of the LAP peptide in mammary alveolar tissue. When the mammary gland was immunostained with LAP antibody, epithelial cells of both infected and non-infected alveoli were immunopositive. These results indicate that LAP is localized in the epithelium of non-infected as well as infected alveolus in the mammary gland in cows.     

Effects of mosapride, a 5-hydroxytryptamine 4 receptor agonist, on electrical activity of the small intestine and cecum in horses

OBJECTIVE: To examine the effects of various doses of mosapride, a 5-hydroxytryptamine 4 receptor agonist, on motility of the small intestine and cecum in horses by use of electrical activity and to determine the dose that provides the optimal response. ANIMAL: 6 healthy adult Thoroughbreds. PROCEDURE: Electrical activity of the small intestine and cecum was recorded before and after mosapride administration by use of an electrogastrograph. Mosapride (0.5, 1, 1.5, and 2 mg/kg) was dissolved in 200 mL of water and administered orally to horses through a nasogastric tube. Three hours after drug administration, mean amplitude of electrical activity calculated for a period of 30 minutes was expressed as the percentage of the mean amplitude of electrical activity for a period of 30 minutes before drug administration. RESULTS: Mosapride administered orally increased the percentage of the mean amplitude of electrical activity in the small intestine and cecum in a dose-dependent manner. Mean +/- SD values differed significantly for 1, 1.5, and 2 mg/kg (127.0 +/- 12.5%, 137.7 +/- 22.2%, and 151.1 +/- 24.0%, respectively) in the small intestine and for 1.5 and 2 mg/kg (130.1 +/- 34.5% and 151.6 +/- 45.2%, respectively) in the cecum. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of results of this study clearly documents that mosapride promotes motility in the small intestine and cecum of horses and that the optimal orally administered dosage is 1.5 to 2 mg/kg. Therefore, mosapride may be useful for treatment of horses with gastrointestinal tract dysfunction.     

Successful use of an acceleration rate response pacemaker with a transvenous steroid-eluting screw-in lead for third-degree atrioventricular block in a labrador retriever

Permanent pacemakers are commonly used in veterinary practice and can have a dramatic effect on the treatment of heart block. A Labrador Retriever dog suffering from exercise intolerance secondary to third degree atrioventricular block was treated with a new pacemaker system. A steroid-eluting screw-in type lead that has the advantage of being more fixed to the myocardial wall without increasing the pacing threshold was used. The heart rate was regulated with an acceleration sensing pacemaker generator that included several automatic modulation systems. Nineteen months after implantation, the dog has a normal level of activity. The present case suggests that this pacemaker design may offer important advantages for canine patients.     

Effects of fallen snow on the voluntary intake and grazing behavior of Hokkaido native horses in winter woodland with underlying Sasa senanensis

The intake, digestibility, and grazing behavior of Hokkaido native horses were investigated in winter woodland with underlying Sasa senanensis. Twenty Hokkaido native horses were grazed in a woodland (8.52 ha) for 14 days in December (snowy; 20 cm of snow fall) and in November (non‐snowy) of the next year. Three mares (360 kg of average bodyweight) were used in each experiment. The fecal collection, used to investigate the intake and digestibility by the double‐indicator method, and behavioral observations were carried out for 24 h on the 6th day in each experiment. During the snowy and non‐snowy periods, the dry matter intakes of sasa foliage were 7.3 and 9.6 kg/d, respectively, which is 2.1 and 2.7% when expressed as a percentage of bodyweight. The apparent digestibility of nutrients and energy released during the snowy and non‐snowy periods were similar. The proportions of digestible energy intakes to maintenance requirement in the mares were 119 and 164% during the snowy and non‐snowy periods, respectively. Their bodyweight remained constant during the non‐snowy period; however, a slight loss in their bodyweight was observed during the snowy period. The duration of grazing on sasa foliage was 416 and 544 min/d during the snowy and non‐snowy periods, respectively, the daily grazing area of the mares during these periods was 2.0 and 5.1 ha, respectively. In other words, the area during the snowy period was smaller than that during the non‐snowy period (P < 0.05). Fallen snow, even with a depth of approximately 20 cm, inhibited the grazing behavior and intake of sasa foliage by Hokkaido native horses grazing in winter woodland pasture.     

Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

The present study was undertaken to develop a novel, practical and simple procedure for enzyme‐linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti‐estrone antibody and horseradish peroxidase‐labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra‐assay and inter‐assay coefficients of variation were 5.3–8.1% and 13.4–15.7%, respectively. The recovery rate was 78–102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.     

苹果自花结实性品种花柱S - 核酸酶基因的克隆

 从苹果二倍体品种‘Ontario’和‘迎秋’花柱中分别克隆到了两个S - 核酸酶基因。同源性比较结果, ‘Ontario’和‘迎秋’都有一个与‘富士’Sf - 核酸酶基因完全相同的序列。‘Ontario’另一个S等位基因编码一个新的S - 核酸酶, 被命名为S8 - 核酸酶; ‘迎秋’另一个S等位基因与苹果‘红茜’的Sd - 核酸酶基因序列相同。