Rapid determination of pathogens in mastitic milk of dairy cows using Gram staining

This study aimed to determine whether causative pathogens in mastitic milk can be determined by Gram staining after the centrifugation of milk. Gram staining was performed using unconcentrated and concentrated milk cells. Using this method, we found that the background of microscopic image of unconcentrated milk cells was complex and bacteria were difficult to detect. In contrast, the background of the smears in the concentrated milk cells was translucent, and bacterial and somatic cells were clearly visible. The sensitivity and specificity of the Gram staining of concentrated milk cells were 84.4% and 86.0% and 50.0% and 94.5% for the detection of gram-positive and gram-negative bacteria, respectively. The presented method provides a simple and inexpensive means of determining mastitis-causing pathogens.     

Mapping QTL for fat area ratios and serum leptin concentrations in a Duroc purebred population

The reduction of extra subcutaneous, intermuscular and abdominal fat is important to increase the carcass lean percentage of pigs. Image analyses of fat area ratios were effective for estimation of separated fat in pig carcasses. Serum concentrations of leptin are useful as physiological predictors of fat accumulation in pigs. The objectives of the present study were to perform a quantitative trait locus (QTL) analysis for fat area ratios and serum leptin concentrations in a Duroc purebred population. Pigs (n = 226 to 538) were measured for fat area ratios of carcass cross‐sections at the fifth to sixth thoracic vertebrae, half body length and last thoracic vertebra using an image analysis system, and serum leptin concentration. In total, animals were genotyped for 129 markers and used for QTL analysis. For fat area ratios, four significant and 12 suggestive QTLs were detected on chromosomes 1, 6, 7, 8, 9, 12 and 13. Significant QTLs were detected on the same region of chromosome 6, which was located near a leptin receptor gene. For serum leptin concentrations, two significant and two suggestive QTLs were detected on chromosomes 6, 9, and 16, and the QTLs on chromosome 6 were also in the same region for fat area ratios.     

Determination of silver in soils by atomic absorption spectrometry

A method for the determination of Ag in soils using atomic absorption spectrometry is described. The method involves the extraction of Ag from soil by boiling with 6 M HC1 followed by separation of the extracted Ag into methylisobutylketone (MIBK) using sodium N, N-diethyldithiocarbamate (DDTC) as a complexing agent. Silver is determined in the MIBK by direct aspiration into a flame atomic absorption spectrophotometer. The detection limit (S/N=2) for this method is 0.0001 mg L^?1 for aqueous solution and 0.002 mg kg^?1 for soil. The Ag content of even unpolluted soils can be determined by this method. The determination of Ag using this method was shown to be unaffected by the presence of various ions in the soil. The method was able to recover nearly 100% of Ag added to soil and approximately the same amounts of soil Ag were determined using this method as with HF-H₂SO₄ decomposition. For 3 reference soils of the Canadian Certified Reference Materials Project (CCRMP), the Ag values obtained by this method were the same as the values determined by Ebarvia et al. (1988). The amounts of Ag in the soils sampled in the Ichi River basin and the Ichi River sediments were determined using this method. This area has been polluted by Cd, Cu, Pb, and Zn discharged from the Ikuno Mine and Smelter. The Ag values ranged from 0.27 to 6.89 mg kg^?1 which were much higher than the values of the unpolluted soils.     

Effects of the feeding of wild Yeso sika deer (Cervus nippon yesoensis) on the prevention of damage due to bark stripping and the use of feeding sites

Feeding sites for wild Yeso sika deer around Lake Akan, Japan, were established. Effects on the number of deer using the feeding sites, the prevention of bark stripping damage, the amount of feeding, and eating time in a 5‐year period (1999–2003) were evaluated. The number of deer using feeding sites increased with years during the feeding period. The damaged tree ratio after the initiation of feeding markedly decreased compared with 16.5% before the initiation of feeding. After the start of feeding, there were no trees with damage the entire circumference. According to tree species, the number of damaged trees of Ulmus laciniata Mayr as a percentage of all investigated trees was high (5.2%). The total amount of beet pulp feeding increased with the feeding year, showing 4.5‐fold increase. At feeding sites in deer culling, eating behavior was observed during the night. The preventive effects on bark stripping damage continued during the 5‐year feeding period. However, with the course of feeding years, the number of deer using feeding sites and the level of feeding increased.     

Functional Analysis of an ATP-Binding Cassette Transporter Gene in Botrytis cinerea by Gene Disruption

The BMR1 gene encoding an ABC transporter was cloned from Botrytis cinerea. To examine the function of BMR1 in B. cinerea, we isolated BMR1-deficient mutants after gene disruption. Disruption vector pBcDF4 was constructed by replacing the BMR1-coding region with a hygromycin B phosphotransferase gene (hph) cassette. The BMR1 disruptants had an increased sensitivity to polyoxin and iprobenfos. Polyoxin and iprobenfos, structurally unrelated compounds, may therefore be substrates of BMR1. Received 18 December 2000/ Accepted in revised form 18 April 2001     

Some Physico-chemical characteristics of soils influenced by Taal Volcano in reference to soil productivity

Soils of the northwestern part of the Taal Volcano in the Philippines representing four geomorphological units (upper, middle, and lower slopes and alluvial plains) were investigated and related to soil productivity. Results revealed that the soils on the upper and middle slopes contained higher amounts of organic matter and available P and displayed a low P retention together with more favorable physical properties such as loamy soil texture, loose and friable and well-drained soils compared to those on the lower slopes and in the alluvial plains. Due to these favorable soil characteristics, sustained agricultural production was higher at the upper elevations than at the lower elevations. Year-round multistorey / mixed cropping systems of cultivation in the upper and middle landscapes were also made possible because the higher precipitation was evenly distributed coupled with cooler temperatures compared the conditions on the lower slopes and in the alluvial plains. On the other hand, the soils on the lower slopes and in the alluvial plains had a clayey texture and contained a lower amount of organic matter and available P, in addition to the lower precipitation, resulting in reduced land utilization, as indicated by the limited types of crops grown and lower yield of crops.     

Nodulation and competitiveness of gusA-marked Bradyrhizobium japonicum A1017 in soybean

Attempts to improve the symbiotic nitrogen fixation with effective (Brady) rhizobium strains do not always succeed under field conditions due to the lower nodulation competitiveness of the introduced strains than that of the indigenous rhizobia (Triplett and Sadowsky 1992). An introduced strain needs to be marked for monitoring its nodule occupancy under competitive nodulation conditions.     

Isolation and partial characterization of endophytic diazotrophs associated with Japanese sugarcane cultivar

Endophytic bacteria were isolated from the juice of a 6 month-old sugarcane cv. NiF-8, which was collected from Miyako, Okinawa and planted in a field lysimeter in Tsukuba, Ibaraki, Japan. The most probable number of N₂-fixing endophytes was 4.5 × 10⁵ cells per gram of fresh weight sample and 21 isolates have a positive reaction for ARA in an N₂-free semi-solid medium with 10% crystallized sugarcane sugar and 0.5% sugarcane juice. Analyses of some of the biochemical properties of the N₂-fixing isolates indicated that 13 isolates were putative strains of Acetobacter diazotrophicus, 4 isolates showed similar characteristics to those of Herbaspirillum seropedicae, and 4 isolates consisted of Herbaspirillum rubribalbicans-like strains. This study confirmed the existence of N₂-fixing endophytic bacteria in the Japanese sugarcane cv. NiF-8.     

Serological evidence of influenza A virus infection in Kuril harbor seals (Phoca vitulina stejnegeri) of Hokkaido, Japan

For proper management and conservation of the Kuril harbor seal (Phoca vitulina stejnegeri) through disease control, serological analysis was performed for influenza A virus infection in free-ranging seals in Hokkaido, Japan. Serum samples were collected from seals at Nosappu (231 seals), Akkeshi (16) and Erimo (75), between 1998 and 2005, and were analyzed by ELISA. Antibodies to the influenza A virus were detected only in seals from Nosappu. The incidences were 11% (1/9), 3% (2/66), 12% (7/59) and 6% (5/77) in 1998, 2003, 2004 and 2005, respectively. These suggest sporadic infection. Because antibody-positive seals included juvenile seals in each year, the infections were considered to have been circulated since no later than the late 1990s until recent years. ELISA-positive sera were analyzed by hemagglutination inhibition (HI) tests to determine the subtypes. Antibodies to the H3 and H6 subtypes were detected in 10 and 2 sera, respectively. Two of the sera that had antibodies to the H6 subtype also had antibodies to the H3 subtype. These two seals were considered to have been infected with both the H3 and H6 subtypes. This is the first investigation to find antibodies to the H6 subtype in seals. Although the H6 subtype had been isolated only from avians, genetic analysis had suggested that the H6 subtype could become a novel mammalian pathogen. For definitive diagnosis, detection of the virus from the tissue or mucus of seals is required.