Effects of carbon and nitrogen amendment on soil carbon and nitrogen mineralization in volcanic immature soil in southern Kyushu,Japan

Soil N mineralization is affected by microbial biomass and respiration, which are limited by available C and N. To examine the relationship between C and N for soil microbial dynamics and N dynamics, we conducted long-term laboratory incubation (150 days) after C and N amendment and measured changes in C and N mineralization, microbial biomass C, and dissolved C and N throughout the incubation period. The study soil was volcanic immature soil from the southern part of Japan, which contains lower C and N compared with other Japanese forest soils. Despite this, the area is covered by well-developed natural and plantation forests. Carbon amendment resulted in an increase in both microbial biomass and respiration, and net N mineralization decreased, probably due to increasing microbial immobilization. In contrast, N amendment resulted in a decrease in microbial respiration and an increase in net N mineralization, possibly due to decreased immobilization by microbes. Amendment of both C and N simultaneously did not affect microbial biomass and respiration, although net N mineralization was slightly increased. The results suggested that inhibitory effect on microbial respiration by N amendment should be reduced if carbon availability is higher. Thus, soil available C may limit microbial biomass and respiration in this volcanic immature soil. Even in immature soil where C and N substrate is low, soil C, such as plant root exudates and materials from above- and belowground dead organisms, might help to maintain microbial activity and N mineralization in this study site.     

Artificial neural network modeling for predicting final moisture content of individual Sugi (Cryptomeria japonica) samples during air-drying

Sugi (Cryptomeria japonica D. Don) lumber is known to have a large variability in final moisture content (MC_f) and is difficult to dry. This study assessed the capability of artificial neural networks (ANNs) to predict the MC_f of individual wood samples. An ANN model was developed based on initial moisture content, basic density, annual ring orientation, annual ring width, heartwood ratio and lightness (L ^* in the CIE L ^* a ^* b ^* system). The performance of the ANN model was compared with a principal component regression (PCR) model. The ANN model showed good agreement with the experimentally measured MC_f with a higher correlation coefficient (r) and a lower root mean square error (RMSE) than the PCR model, demonstrating the importance of nonlinearity of the variables and the higher capability of the ANN model than the PCR model. By adding redness (a ^* ) and yellowness (b ^* ) and drying time to the input variables of ANNs, r and RMSE values were improved to 0.98 and 1.2 % for the training data set, and 0.85 and 2.2 % for the testing data set, respectively. Although the developed ANNs are available under the limited conditions of this study, our results suggest that the ANNs proposed offer reliable models and powerful prediction capability for the MC_f, even though wood properties vary considerably and their complex interrelations are not fully elucidated.     

Antihyaluronidase action of ellagic acid effectively prevents polyspermy as a result of suppression of the acrosome reaction induced by sperm-zona interaction during in vitro fertilization of porcine oocytes

The present study was conducted to examine the effects of three tannin relatives (tannic acid, TA; gallic acid, GA; and ellagic acid, EA) on antihyaluronidase and reactive oxygen species (ROS) scavenging activity, in vitro fertilization (IVF) parameters, and the acrosome reaction (AR) induced by sperm-zona interaction. Among the three tannin relatives, TA and EA showed the strongest potency for blocking the hyaluronidase activity of boar sperm, with concentration-dependent inhibition over the range of 2-10 microg/ml. In contrast, ROSs were effectively scavenged by TA and GA, but not EA. When cumulus-free oocytes were inseminated in IVF medium containing 5 microg/ml of the tannin relatives, polyspermy was significantly reduced by TA and EA (32 and 29%, respectively) compared with oocytes treated with or without GA (51 and 69%, respectively) under conditions that maintained a high sperm penetration rate (P<0.05). Interestingly, induction of the AR by treatment of preincubated sperm with progesterone was blocked by TA and GA as a result of their higher levels of ROS scavenging activity, while EA, which possessed weak ROS scavenging activity, did not disturb induction of the AR with progesterone. However, the incidence of AR induced by sperm-zona interaction was significantly decreased by the strong antihyaluronidase actions of TA and EA compared with that in the absence of these compounds. Treatment with the compounds caused neither a protective proteolytic modification of the zona pellucida matrix before fertilization nor a reduction in acrosomal proteolytic activity or the number of zona-bound sperm. These findings suggest that the antihyaluronidase action of EA effectively prevents polyspermy by suppression of AR functionality induced by sperm-zona interaction and that hyaluronidase intervention is therefore required during porcine IVF.     

Experimental and clinical studies on plasma leptin in obese dogs

Leptin is a protein synthesized and secreted primarily by adipocytes, and the circulating leptin concentration is elevated in obese humans and rodents. Recently, we have established a sandwich enzyme-linked immunosorbent assay for canine leptin. In the present study, plasma leptin concentrations were measured in experimentally developed obese beagles and in clinically obese dogs. When 5 male beagles were given a high-energy diet for 3 months, all of them became obese and the plasma leptin concentration significantly increased from 2.4+/-1.2 to 4.9+/-0.9 ng/ml, positively correlating with body fat content estimated by the deuterium oxide dilution method (r=0.87). The leptin concentrations of plasma samples collected from 59 dogs in veterinary practices were compared with their body condition scores (BCS). The plasma leptin concentrations of obese dogs were 9.7+/-0.7 and 12.3+/-1.5 ng/ml at BCS=4 and BCS=5, respectively, which were significantly higher than those of optimal (BCS=3) dogs (2.7+/-0.3 ng/ml). There was no significant effect of sex and breed. A weak positive correlation (r=0.37) was found between the plasma leptin concentration and age, probably due to the lesser content of visceral fat in puppies younger than 1 year old. These results indicate that plasma leptin is a good index of adiposity in dogs regardless of breed, age and sex, and may be useful for quantitative assessment of obesity in small animal practice.     

Labeled avidin-biotin enzyme-linked immunosorbent assay for detecting antibody to infectious laryngotracheitis virus in chickens

A labeled avidin-biotin enzyme-linked immunosorbent assay (LAB-ELISA) for detecting antibody to infectious laryngotracheitis (ILT) virus in chicken sera was developed and compared with ordinary ELISA. Purified ILT virus, biotin-labeled anti-chicken IgG rabbit IgG conjugate, and horseradish-peroxidase-labeled avidin were used in the LAB-ELISA. When sera from farm chickens were tested by serum neutralization (SN) and two kinds of ELISA, the correlation rate between SN and LAB-ELISA was 50/50 (100%), and that between SN and ordinary ELISA was 39/50 (78%). In LAB-ELISA, all of the sera that were antibody-negative by SN had low absorbance (A) values (below 0.05), and the A values were closely correlated with the SN indexes. In ordinary ELISA, however, the sera antibody-negative by SN had various A values ranging from 0.06 to 0.32. LAB-ELISA had much lower nonspecific reactions than ordinary ELISA against sera from ILT-negative chickens, even when chickens were 30 weeks old. ILT antibody production after ILT vaccination could be detected by LAB-ELISA. A values peaked 5 weeks postinoculation and were maintained for 17 weeks.     

Prevention of adverse reactions following milbemycin D administration to microfilaremic dogs infected with Dirofilaria immitis

Some adverse reactions such as shock-like reaction and dirofilarial hemoglobinuria (caval syndrome) occasionally occurred in microfilaremic dogs following milbemycin D (Milbe) administration. This study was carried out to seek the prevention of these adverse reactions. In two groups containing 16 and 9 dogs respectively which were administered either chlorpheniramine maleate (1 mg/kg) or indomethacin (2.5 mg/kg) simultaneously with Milbe (1 mg/kg), the incidence of clinical signs such as the pale color of the visible mucous membranes, respiratory disorders, caval syndrome and shock-like reaction as well as changes in clinical parameters such as RBC and WBC counts, WBC profile and serum total protein, were almost equal to that observed in the group administered Milbe alone. In 41 dogs administered prednisolone (1 mg/kg) simultaneously with Milbe (1 mg/kg), no shock-like reaction was observed. Changes in clinical parameters were different from those in the group administered Milbe alone, whereas some clinical signs of adverse reactions, including caval syndrome, were observed. These results indicated that prednisolone was effective for prevention of the shock-like reaction in microfilaremic dogs induced by Milbe.     

The Culture of the Rotifer Brachionus plicatilis with Chlorella vulgaris Containing Vitamin B12 in its Cells

The rotifer Brachionus plicatilis requires vitamin B₁₂. Freshwater Chlorella , which is produced by traditional culture, cannot support rotifer growth under bacteria-free conditions. However, Chlorella enriched with vitamin B₁₂ can support rotifer growth. To attain stable mass production of Brachionus , it is desirable to develop a food that can completely support growth of the rotifers. The authors cultured rotifers at experimental and mass culture scale with concentrated Chlorella vulgaris suspension enriched with vitamin B₁₂ in their cells.
Chlorella suspensions were prepared containing different amounts of vitamin B₁₂ in their cells, and rotifers were then cultured in 20 ml of the prepared suspensions. The highest rotifer yield was obtained from the group cultured with Chlorella containing more vitamin B₁₂ in their cells. The suitable content of vitamin B₁₂ in the concentrated Chlorella suspension commercially available for mass culture of the rotifer is considered to be 200 _μg per 100 g dry matter of Chlorella . The amount of vitamin B₁₂ necessary to produce one individual rotifer is calculated at 0.32 pg.
The authors conducted mass production of the rotifer with baker's yeast and refrigerated concentrated Chlorella containing vitamin B₁₂ Rotifer culture with vitamin B₁₂ was more stable and showed 1.3 times higher production than with normal Chlorella .     

Correlated responses of respiratory disease and immune capacity traits of Landrace pigs selected for Mycoplasmal pneumonia of swine (MPS) lesion

Five generations of Landrace pigs selected for average daily gain, backfat thickness, Mycoplasmal pneumonia of swine (MPS) lesion score, and plasma cortisol levels, was executed to decrease the MPS lesion score. Genetic parameters and correlated genetic responses for respiratory disease and peripheral blood immune traits were estimated in 1395 Landrace pigs. We estimated the negative genetic correlation of MPS lesion score with phagocytic activity (PA) at 7 weeks of age (‐0.67). The breeding values of PA at 7 weeks of age and 105 kg body weight and the correlated selection response of the ratio of granular leukocytes to lymphocytes at 105 kg body weight were significantly increased, and sheep red blood cell‐specific antibody production (AP) was significantly decreased in a selection‐dependent manner. Increasing of natural immunological indicators (e.g. PA) and decreasing of humoral immunological indicator (e.g. AP) were observed due to genetically decreasing MPS lesion score.     

Fate of prothiofos (O-2,4-dichlorophenyl O-ethyl S-propyl phosphorodithioate) in rats

The fate of prothiofos (O-2,4-dichlorophenyl O-ethyl S-propyl phosphorodithioate) were studied in male rats 1, 3, 8, 24, 48, and 120 hr after oral administration. In the determination of prothiofos by gas chromatography-mass spectrometry, pentadeuteroethoxy-labeled prothiofos was used as the internal standard and the carrier of residual prothiofos. Prothiofos was rapidly absorbed from the small intestine and had substantially disappeared from the gastrointestinal tract within 24 hr after dosing. The residual concentrations of prothiofos in all organs analyzed, reached a maximum at 3 hr after dosing, and then diminished exponentially. The major urinary metabolites were 2,4-dichlorophenol (21% of administered prothiofos), its conjugated substances (26%), O-2,4-dichlorophenyl O-ethyl hydrogen phosphorothioate (14%), and O-2,4-dichlorophenyl O-ethyl hydrogen phosphate (25%). While prothiofos was found in the feces (3.8%), prothiofos oxygen analog (prothiofos-oxon) was scarcely detected in any excreta. Results obtained with a single dosing of prothiofos-oxon indicated that the oxygen analog formed from prothiofos in vivo was rapidly degraded through cleavage of the PS bond and the liberation of 2,4-dichlorophenol. The low mammalian toxicity of prothiofos is probably due to depropylthiolation in the molecule.