A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle

A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in cultured milk samples were detected by this simplified PCR-based method as well as a standard PCR technique. The minimum concentration limit for detecting mycoplasma by the simplified PCR was estimated to be about 2.5 × 10³ cfu/mL and was similar to that of the standard PCR. We compared the specificity and sensitivity of the simplified PCR to those of a culture method. Out of 1,685 milk samples cultured in mycoplasma broth, the simplified PCR detected Mycoplasma DNA in 152 that were also positive according to the culture assay. The sensitivity and specificity of the simplified PCR were 98.7% and 99.7%, respectively, for detecting mycoplasma in those cultures. The results obtained by the simplified PCR were consistent with ones from standard PCR. This newly developed simplified PCR, which does not require DNA purification, can analyze about 300 cultured samples within 3 h. The results from our study suggest that the simplified PCR can be used for mycoplasma mastitis screening in large-scale dairy farms.     

Distribution of photoperiod-insensitive alleles Ppd-B1a and Ppd-D1a and their effect on heading time in Japanese wheat cultivars

The genotypes of photoperiod response genes Ppd-B1 and Ppd-D1 in Japanese wheat cultivars were determined by a PCR-based method, and heading times were compared among genotypes. Most of the Japanese wheat cultivars, except those from the Hokkaido region, carried the photoperiod-insensitive allele Ppd-D1a, and heading was accelerated 10.3 days compared with the Ppd-D1b genotype. Early cultivars with Ppd-D1a may have been selected to avoid damage from preharvest rain. In the Hokkaido region, Ppd-D1a frequency was lower and heading date was late regardless of Ppd-D1 genotype, suggesting another genetic mechanism for late heading in Hokkaido cultivars. In this study, only 11 cultivars proved to carry Ppd-B1a, and all of them carried another photoperiod-insensitive allele, Ppd-D1a. The Ppd-B1a/Ppd-D1a genotype headed 6.7 days earlier than the Ppd-B1b/Ppd-D1a genotype, indicating a significant effect of Ppd-B1a in the genetic background with Ppd-D1a. Early-maturity breeding in Japan is believed to be accelerated by the introduction of the Ppd-B1a allele into medium-heading cultivars carrying Ppd-D1a. Pedigree analysis showed that Ppd-B1a in three extra-early commercial cultivars was inherited from ‘Shiroboro 21’ by early-heading Chugoku lines bred at the Chugoku Agriculture Experimental Station.     

Effect of cashew nut shell liquid on metabolic hydrogen flow on bovine rumen fermentation

Effect of cashew nut shell liquid (CNSL), a methane inhibitor, on bovine rumen fermentation was investigated through analysis of the metabolic hydrogen flow estimated from concentrations of short‐chain fatty acids (SCFA) and methane. Three cows were fed a concentrate and hay diet without or with a CNSL‐containing pellet. Two trials were conducted using CNSL pellets blended with only silica (trial 1) or with several other ingredients (trial 2). Methane production was measured in a respiration chamber system, and energy balance and nutrient digestibility were monitored. The estimated flow of metabolic hydrogen demonstrated that a part of metabolic hydrogen was used for hydrogen gas production, and a large amount of it flowed into production of methane and SCFA in both trial 1 and 2, when CNSL was administered to the bovine rumen. The results obtained by regression analyses showed that the effect of CNSL supply on methane reduction was coupled with a significant (P < 0.01) decrease of acetate and a significant (P < 0.01) increase of propionate and hydrogen gas. These findings reveal that CNSL is able to reduce methane and acetate production, and to increase hydrogen gas and propionate production in vivo.     

Polymerase chain reaction-based identity assay for pathogenic turkey Eimeria

Diagnosis of turkey Eimeria infection by conventional parasitologic methods is challenging and, until now, no molecular tools existed that clearly distinguished the four widely recognized pathogenic species: Eimeria adenoeides, E. meleagrimitis, E. gallopavonis, and E. dispersa. In this study, the internal transcribed spacer region one (ITS-1) was amplified and sequenced from 23 conventionally characterized reference samples. Phylogenic analysis segregated samples into five distinct cluster groups. The ITS-1 region(s) within each cluster were of a particular length and shared from 96% to 100% identity, while amplified ITS-1 region(s) between clusters differed in length and shared only 10.6% to 49.7% sequence identity. In addition, we developed PCR primer sets as diagnostic tools capable of specifically identifying members of each of the five clusters.     

Comparative studies on lipid profiles and amino acid composition of wild and cultured Dojo loach Misgurnus anguillicaudatus obtained from southern Japan

The aim of this study was to determine the differences in the nutritional properties of wild and cultured loach Misgurnus anguillicaudatus obtained from southern Japan. Major parameters, such as whole-body proximate composition, lipid classes, and fatty acid and amino acid composition, were investigated. The whole-body total lipid content of cultured loach was twofold higher than that of wild loach and the proportion of neutral lipids was significantly higher in cultured loach than in wild loach. However, the polar lipid content was two- to threefold lower in cultured loach than in wild loach. Compared to the wild loach, higher contents of 20:5n-3 and 22:6n-3 were detected in cultured loach, but much lower contents of 20:4n-6. In terms of whole-body amino acid composition, wild and cultured loach showed similar profiles. In conclusion, the consumption of cultured loach will contribute to dietary n-3 highly unsaturated fatty acids intake, with a benefit to human health.     

Effects of palm oil blended with oxidized fish oil on growth performances, hematology, and several immune parameters in juvenile Japanese sea bass, Lateolabrax japonicas

A 60-day feeding trial was conducted to determine the effects of palm oil blended with oxidized and non-oxidized fish oil on growth performances, hematology, and non-specific immune response in juvenile Japanese sea bass, Lateolabrax japonicas. Japanese sea bass (1.73 ± 0.01 g) were fed seven experimental diets containing 100 g/kg of dietary lipid in forms of palm oil (10P), fish oil (10F), fish oil blended with palm oil at different ratios, 6:4 (6F4P) and 4:6 (4F6P), oxidized fish oil (10OF), and oxidized fish oil blended with palm oil at different ratios, 6:4 (6OF4P) and 4:6 (4OF6P). After the feeding trial, the following results were illustrated. No significant effects were observed in survival, feed conversion ratio, condition factor, and hematocrit after feeding with experimental diets for 60 days. The relatively higher specific growth rate and hematology were observed in 6F4P. Furthermore, both palm oil and oxidized fish oil acted as a negatively on serum lysozyme activity (P < 0.05). This study suggested that a ration of 6F4P is recommended as an innocuous ratio for Japanese sea bass. Furthermore, according to the present investigation, palm oil seems to have the ability to improve the protein efficiency when added to oxidized fish diets as well as a positive trend to the growth performance (P > 0.05).     

Optimization model for cropping-plan placement in paddy fields considering agricultural profit and nitrogen load management in Japan

An optimization model for cropping-plan placement on field plots is presented for supporting decision-making on agricultural management by a farming organization. The mixed 0?C1 programming technique is employed to select the next planting crop at each field plot in a holistic manner. Reduction of total nitrogen discharged from field plots to the downstream end of the drainage canals is expressed as an objective function of the model to balance an achievement of economic goal and environmental conservation. Some Japanese governmental policies on regulating rice cropping areas and on promoting production of particular upland field crops can be formulated in the model. A computational example of cropping-plan placement on field plots managed under integrated policies is given by operating the optimization model with various weights associated with the objectives. The procured trade-off curve and corresponding patterns of cropping-plan could be useful in the decision-making by the farming organization.     

Radiotherapy-induced tumor lysis syndrome in a dog with thymoma

A 13-year-old, female, mixed-breed dog with a huge cranial mediastinal mass underwent radiotherapy (RT). On the following day, the dog presented with lethargy and anorexia. Hematological examination revealed elevated levels of blood urea nitrogen, creatinine, inorganic phosphorus, potassium, lactate dehydrogenase, creatine phosphokinase and aspartate aminotransferase, decreased calcium level, and metabolic acidosis. Urine output markedly decreased. The patient recovered with fluid therapy and diuretic therapy; however, died suddenly from an unknown cause 11 days after RT completion. Histopathological examination after necropsy showed thymoma in the cranial mediastinum and extensive tubular necrosis of both kidneys which may be due to RT-induced tumor lysis syndrome (TLS). This report suggests that the risk of TLS should be evaluated in dogs with thymoma who undergo RT.     

Effect of dietary lipid oxidation with vitamin C and E supplementation on fillet quality of red sea bream,Pagrus major (Temminck & Schlegel) during storage

An experiment was conducted to determine the effects of different levels of dietary vitamin C (VC) and E (VE) supplementation on fillet quality of red sea bream fed oxidized fish oil (OFO). Fish with an average body weight of 205.0 g were fed four test diets for 9 weeks. Control diet contained fresh fish oil (FFO) with 100 mg kg^?1 of VE and 500 mg kg^?1 of VC (FFO100E/500C). The other three diets contained OFO with varying levels of VE (mg kg^?1) and VC (mg kg^?1) (OFO100E/500C, OFO200E/500C and OFO200E/1000C). After feeding trial, two fillets from each fish by hand filleting were stored in a refrigerator at 4°C for 96 h during analyses. Results showed that fish fed OFO increased fillet thiobarbituric acid reactive substances (TBARS) and K‐value, and decreased fillet VC and VE concentrations during storage time. Supplementation of VC did not have any detectable effect on fillet quality. Increasing dietary VE supplementation increased fillet VE concentrations, reduced fillet TBARS and K‐value values of red sea bream. Therefore, we suggest that dietary supplementation of 200 mg kg^?1 of vitamin E could improve fillet oxidative stability of red sea bream fed OFO.     

Relationship between the temperature and the overwintering of water lettuce (Pistia stratiotes) at Kowataike,a branch of Yodogawa River,Japan

The successful overwintering of water lettuce (Araceae, Pistia stratiotes L.) is reported in Kowataike, a pond in Kyoto Prefecture, Japan, in the temperate climate zone. Overwintering was observed in the northern zone of Kowataike, where warm water discharge flows from an upstream electrical appliance factory. From December 2007 to January 2008, several overwintering water lettuce plants were observed in the northern zone where the water temperature was abnormally high (average: 21.9°C; low: 19.5°C). Water in the central zone of Kowataike was isolated from the warm water discharge. The temperatures here were much lower (average: 8.2°C; low: 4.2°C) and no water lettuce plant survived here. The overwintering in the northern zone fluctuated annually. This fluctuation appeared to be correlated with the period of warm water discharge, which varied from year to year and resulted in higher water temperatures most, but not all, of the winter. The results suggest that artificial environmental factors, such as anthropogenic warm water discharge, could enable water lettuce to overwinter. In the northern zone of Kowataike under the influence of the warm water discharge, only small water lettuce plants overwintered. This may be attributed to the presence of older plants with larger leaves that seemed to protect the small rosettes from frost. Second, the air warmed by the warm discharge accumulated in the space under the piles of dead large leaves, which created conditions similar to those of a greenhouse. These two mechanisms apparently allow the small water lettuce plants to be surrounded by moderate atmospheric conditions.