Application of ELISA-based kit for detecting AOZ and determining its clearance in eel tissues

Furazolidone, an antibacterial drug that was once widely used in the livestock industry and aquaculture, is now prohibited in numerous countries. It is difficult to detect residual furazolidone because it is readily metabolized in animal tissues but, by using and liquid chromatography coupled with tandem mass spectrometry, its metabolite, 3-amino-2-oxazolidinone (AOZ) can be detected. Here we describe the validity of an enzyme-linked immunosorbent assay (ELISA) kit to detect AOZ in Japanese eel Anguilla japonica tissue. ELISA is capable of detecting AOZ at 1.0 μg/kg in an eel sample with excellent accuracy and precision. Our results show that ELISA is suitable for regulatory purposes and for studying the fate of AOZ residues in eel treated with furazolidone. To measure the persistence of AOZ in eel tissues, eels (1.4–6.5g) were immersed in tanks containing 2 and 10 mg furazolidone/L for 3 h, and then maintained in a tank supplying well water for the next 160 days. The half-lives of AOZ, calculated from the linear terminal part of the excretion curve, were 25.0 days in muscle and 21.6 days in liver from fish exposed to 2 mg/L furazolidone. In the eels treated with 10 mg/L furazolidone, by contrast, high levels of AOZ were detected in liver and muscle, but the half-lives of AOZ were similar to those in fish treated with 2 mg/L furazolidone. The half-lives of AOZ in eel tissues were prolonged by the condition of low water temperature.     

Loss of and damage to scallops due to storms in the Sea of Okhotsk

In the coastal zone of the Sea of Okhotsk off Hokkaido, Japan, the giant ezo scallop ( Patinopecten yessoensis ) is widely cultivated on the sea bed, making its fishery a major industry. However, the area experiences intense cyclones, including typhoons, about once a decade and the scallop resources appear to have received severe damage from the storms. Such an event occurred on 29 October 1988. The storm caused 12-m-high wind waves, which appeared to be fully developed sea waves. The water above the sea bed repeatedly moved back and forth at speeds of 1∼2 m s^-1 causing damage to the scallops due to collisions and rubbing with gravel and other scallops. From a comparison of wind data with the annual catch of scallops, intense cyclones seem to play an important role on the year-to-year fluctuations in the scallop fishery.     

Effects of heat stress on the endometrial epidermal growth factor profile and fertility in dairy cows

The endometrial epidermal growth factor (EGF) profile is an indicator of uterine function and fertility in cattle. The present study aimed to investigate the effects of heat stress on the endometrial EGF profile and fertility in lactating Holstein cows. The endometrial EGF profiles of 365 cows in the Hokkaido and Kyushu regions were examined between June and September (heat stress period, n = 211) and between October and January (control period, n = 154). EGF profiles were investigated using uterine endometrial tissues obtained by biopsy 3 days after estrus (Day 3). The proportion of cows with an altered EGF profile was higher between June and September than between October and January (41.2 vs. 16.2%, P < 0.05). The effects of rectal temperature on Days 0 and 3 on the endometrial EGF profile were also assessed in cows (n = 79) between June and September in the Kyushu region. A single embryo was transferred to cow on Day 7 to evaluate fertility (n = 67). Regardless of the rectal temperature on Day 3, the proportion of cows with an altered EGF profile was higher (64.1 vs. 30.0%, P < 0.05) and the pregnancy rate after embryo transfer (ET) was lower (26.7 vs. 51.4%, P < 0.05) in cows with a rectal temperature ≥ 39.5°C on Day 0 than in cows with a rectal temperature < 39.5°C on Day 0. The present results indicate that alterations in the endometrial EGF profile induced by an elevated body temperature on Day 0 contributed to reductions in fertility in lactating dairy cows during the heat stress period.     

Development of X-ray computed tomography for live standing cattle

An X‐ray computed tomography (CT) system for live standing cattle was developed for studying the meat yield, carcass composition and so on. The gantry contained three X‐ray tubes and detectors that corresponded to each X‐ray tube. The system was able to operate while the animal remained standing. The scan area had a diameter of 900 mm. The Musculus longissimus and Musculus trapezius areas, and the back fat thickness in the CT image were evaluated and compared to the actual cross‐section of the carcass using eight cattle. The differences among the muscles, and the subcutaneous and intermuscular fat were easily recognized. The correlation coefficient between the CT image and the actual carcass photograph of the M. longissimus area and the back fat thickness was high (r = 0.84, r = 0.93, P < 0.01). The present study demonstrated that muscle, fat and bone can be clearly imaged from a live standing animal using X‐ray equipment.     

Concentrations and specific antibodies to staphylococcal enterotoxin-C and toxic shock syndrome toxin-1 in bovine mammary gland secretions,and inflammatory response to the intramammary inoculation of these toxins

To investigate the pathological role of staphylococcal enterotoxins (SEs) and toxic shock syndrome toxin-1 (TSST-1) in bovine mastitis, the production of SEs and TSST-1 was investigated in staphylococci isolated from 120 mammary gland secretions (MGS, 51 from no clinical sign-mammary glands and 69 from staphylococcal mastitic-mammary glands) collected from dairy farms where staphylococcal mastitis frequently occurred in Miyagi and Yamagata prefectures from 1997 to 1998. Concentrations of these toxins and specific antibody titers in each MGS were also measured. Furthermore, SEC and TSST-1 were inoculated into lactating mammary glands and inflammatory responses were analyzed. A high percentage of staphylococci including Staphylococcus aureus and coagulase-negative staphylococci isolated from both no clinical sign- and mastitic-MGS produced both SEC and/or TSST-1. The concentration of SEC increased with the severity of the mastitis, and was significantly higher (P<0.05) in acute mastitic-than in no clinical signs-MGS. Titers of specific antibodies to TSST-1 in MGS were significantly higher (P<0.05) than those to SEC, regardless of whether or not the cows were lactating or mastitic. Specific antibodies purified from MGS neutralized each toxin in vitro. A significant increase (P < 0.05) in somatic cell counts was induced by the intramammary inoculation of SEC but not TSST-1. These findings indicated that SEC rather than TSST-1 plays an important role in the pathology of staphylococcal bovine mastitis. The inflammatory activity of TSST-1 was probably neutralized by specific antibodies in MGS.     

性别控制胚胎玻璃化一步法稀释冷冻保存技术的研究

为了创立在用户庭院就能应用的玻璃化保存性别控制胚胎的冷冻剂稀释处理方法，采用细管内3种不同液体层构成（A、B、C）法，对新鲜分割胚一步法稀释后的存活能力进行了研究；其次用特定细管最小容量冷却法与上述3种不同液体层构成中效果较好的B法对玻璃化冷冻溶解的新鲜分割胚、体外受精分割胚及冷冻溶解的体外受精分割胚进行一步法稀释后的存活能力予以对比。结果，3种不同液体层构成间胚胎的存活能力无显著差异，B法较高；应用特定细管最小容量冷却法在冷冻溶解的新鲜分割胚、体外受精分割胚及冷冻溶解的体外受精分割胚一步法稀释后的存活能力均显著高于B法。结果表明，特定细管最小容量冷却法是更有效的保存方法。     

Optimal application timing of simeconazole granules for control of rice kernel smut and false smut

We investigated the optimal timing of simeconazole (RS-2-(4-fluorophenyl)-1-(1H-1,2,4-triazol-1-yl)-3-trimethylsilylpropan-2-ol) application for controlling rice kernel smut in field trials in Miyagi Prefecture, Japan, using formulations of simeconazole (1.5% granules). The field tests revealed that a submerged application of simeconazole granules (450–600 g ai/ha) at 1–5 weeks before heading was highly effective against kernel smut, with treatments 1–2 weeks before heading being the most effective. Submerged application of the fungicide at 2–5 weeks before heading was also highly effective against false smut, with treatment 3 weeks before heading being the most effective. These periods overlap the timing for optimal application of simeconazole to control rice sheath blight and ear blight. Consequently, we concluded that treatment with simeconazole 2–3 weeks before heading can be a useful tool for controlling all four diseases.     

Lesion-based analysis of leaf blast suppression in mixture of rice cultivar and a resistant near-isogenic line

Leaf blast suppression in multilines was evaluated based on the number of susceptible lesions observed in a pure stand of susceptible rice cultivar Sasanishiki, and in 1 : 1 and 1 : 3 mixtures of Sasanishiki and a resistant near-isogenic line, Sasanishiki BL4 or BL7, from 1998 to 2001. The number of lesions first observed in fields in the 1 : 1 and 1 : 3 mixtures were close to theoretical numbers calculated using the number of lesions observed in the pure stands and the ratios of the susceptible Sasanishiki in the mixtures. The ratio of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand was 0.29 and 0.09, respectively. The relationship between these ratios and the ratios of susceptible Sasanishiki in mixtures was defined in an equation to estimate the degree of leaf blast suppression. Validation studies for the ratios of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand were conducted in two different locations and showed that the ratios are almost acceptable. The calculated autoinfection to alloinfection ratio was 1.3 and 1.4 in the 1 : 1 and 1 : 3 mixtures, respectively, suggesting that the calculated ratio will affect the degree of leaf blast suppression. Thus, predictors were obtained to estimate leaf blast suppression for effective blast control in multilines.     

Isolation and characterization of lectins from the AG-D group of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> species

Isolates from 18 anastomosis groups (AGs) of binucleate Rhizoctonia were screened for lectin activity. Eight AGs (AG-B, AG-D, AG-F, AG-G, AG-H, AG-I, AG-R and AG-U) had low to moderate lectin activities. Among these, members of AG-D and AG-I had the highest activity. Partially purified lectins from AG-D preferentially agglutinated human blood type A to type B and O. Mucin and galactose were the most potent inhibitors among the tested carbohydrates. The molecular masses of these lectins ranged from 12.7 kDa for the monomer to 62 kDa for the pentamer type. Proline, alanine, glutamic acid, aspartic acid, leucine, threonine, serine and tyrosine were the major amino acid components of these lectins. Lectins from AG-D were stable at 4–50°C and from pH 6.0 to 10.0. When assayed with isoelectric focusing, these lectins gave bands at pI 9.30. Specificity of lectins from AG-D to galactose and its derivatives suggest a possible recognition role in this fungal species.