Improvement of preservation method for ammonia-oxidizing bacteria by freeze-drying

Long-term preservation of ammonia-oxidizing bacteria had been achieved until now by using the silica gel-freezing method. Recently, the freeze-drying method for the preservation of microorganisms has been widely used, and has been applied to the preservation of ammonia-oxidizing bacteria. The 10% skim milk-l% pyruvate or glutamate ratio was optimum as a dispersion-medium, and the preservation temperature of the freeze-dried cells was effective at -80°C. This method of preservation was found to be considerably more effective than the silica gel-freezing method.     

Effects of dietary phosphorus level on non-faecal phosphorus excretion from yellowtail (Seriola quinqueradiata Temminck & Schlegel) fed purified and practical diets

Non-faecal phosphorus (P) was determined for large yellowtail to estimate a minimum available P requirement (Experiment  1) and to justify inorganic P supplementation in a fish meal-based diet (Experiment 2). In Experiment 1, purified diets with incremental P concentrations were fed to yellowtail (mean weight 917 g) at a feeding rate of 1.5% of body weight. The peaks of non-faecal P excretion appeared 5–6 h after feeding in fish fed more than 4.5 g available P kg⁻¹ dry diet. Broken-line analysis indicated that the minimum available P requirement was 4.4 g kg⁻¹ dry diet. In Experiment 2, a purified diet (PR) containing 6.5 g available P kg⁻¹ and a fish meal-based diet with (F1) and without (F0) additional phosphorus were fed to yellowtail (mean weight 1.1 kg) at 1.5% (PR) and 2% (F0 and F1) feeding rates respectively. There was no significant difference in P excretion between fish fed the F0 (5.5 g soluble P kg⁻¹ dry diet) and the PR diet. However, significantly higher (34.5%) amounts of non-faecal P excretions (7.4 g soluble P kg⁻¹ dry diet) were found in fish fed F1 compared with the F0 diet. This suggested that there was an excess of dietary P in the F1 diet and that supplementation is not needed in fish meal-based diets for large yellowtail.     

Effect of nucleotides supplementation to low‐fish meal feed on long‐chain polyunsaturated fatty acid composition of juvenile rainbow trout Oncorhynchus mykiss

A feeding trial was conducted to evaluate the effect of nucleotides supplementation to low‐fish meal feed on growth and fatty acid composition of rainbow trout. Six isonitrogenous (42% crude protein) and isolipidic (18% crude lipid) diets were formulated containing fish meal and plant ingredients as main protein sources. The control diet was a basal diet without supplementation of nucleotides, and five experimental diets were prepared by supplementing one of the five different nucleotides in the form of 5′‐monophosphate (0.15%), that is inosine (IMP), adenosine (AMP), guanosine (GMP), uridine (UMP) and cytidine (CMP) onto basal diet. Two hundred forty juvenile rainbow trout with an initial average body weight 9.8 g were randomly distributed into twelve aquaria. After 15 weeks of feeding period, growth performance and feed utilization of rainbow trout were not significantly different among dietary treatments. Dietary GMP, UMP and CMP tended to accumulate crude lipid in the muscle and whole fish body. Moreover, dietary GMP, UMP and CMP significantly increased hepatic 18:3n‐3 and long‐chain homologue 18:4n‐3 and 20:4n‐3 contents. Hepatic 18:2n‐6 content showed also increase in fish fed GMP, UMP and CMP diets, but decreased in long‐chain homologue 20:3n‐6 and 20:4n‐6 contents. Decrease in 20:4n‐6, 20:5n‐3 and 22:6n‐3 contents was also found in the muscle of fish fed IMP, GMP and CMP diets. The present study clearly showed that there was no positive effect of dietary nucleotides on growth of fish, but dietary nucleotides particularly GMP, UMP and CMP altered polyunsaturated fatty acid composition of rainbow trout.     

Heritability and genetic correlation estimates of semen production traits with litter traits and pork production traits in purebred Duroc pigs

We estimated heritabilities of semen production traits and their genetic correlations with litter traits and pork production traits in purebred Duroc pigs. Semen production traits were semen volume, sperm concentration, proportion of morphologically normal sperms, total number of sperm, and total number of morphologically normal sperm. Litter traits at farrowing were total number born, number born alive, number stillborn, total litter weight at birth, mean litter weight at birth, and piglet survival rate at birth. Litter traits at weaning were litter size at weaning, total litter weight at weaning, mean litter weight at weaning, and piglet survival rate from birth to weaning. Pork production traits were average daily gain, backfat thickness, and loin muscle area. We analyzed 45,913 semen collection records of 896 boars, 6,950 farrowing performance records of 1,400 sows, 2,237 weaning performance records of 586 sows, and individual growth performance records of 9,550 animals measured at approximately 5 mo of age. Heritabilities were estimated using a single-trait animal model. Genetic correlations were estimated using a 2-trait animal model. Estimated heritabilities of semen production traits ranged from 0.20 for sperm concentration to 0.29 for semen volume and were equal to or higher than those of litter traits, ranging from 0.06 for number stillborn and piglet survival rate at birth to 0.25 for mean litter weight at birth, but lower than those of pork production traits, ranging from 0.50 for average daily gain to 0.63 for backfat thickness. In many cases, the absolute values of estimated genetic correlations between semen production traits and other traits were smaller than 0.3. These estimated genetic parameters provide useful information for establishing a comprehensive pig breeding scheme.     

Low water exchange Gracilariopsis bailiniae Zhang & B.M. Xia culture in intensive milkfish culture effluents: effects of seaweed density on seaweed production and effluent treatment

This study tested the feasibility of a low-cost seaweed biofiltration system for pond-based aquaculture through an indoor-integrated fish-seaweed culture experiment using weekly nutrient supply regime and different seaweed stocking densities. The culture experiment was conducted in glass aquaria that were stocked with Gracilariopsis bailiniae at 3 densities (low = 0.5 kg m^?2, middle = 2 kg m^?2 and high = 3.5 kg m^?2) and provided either with effluents from intensive milkfish (Chanos chanos) culture or with effluent-free seawater (control) as nutrient source. Stocking density was used as a factor in optimizing nutrient availability for growth and nutrient removal under such low water exchange conditions. Our results showed that G. bailiniae cultivated in milkfish effluents had higher growth, biomass and nitrogen yields than those cultivated in effluent-free seawater. Among the different stocking densities tested, highest growth rate (1.03 % day^?1) was obtained in the middle density. Increasing biomass and nitrogen yields were also obtained at this density until the end of the culture period. Poorer growth rates at low and high stocking densities were attributed to light limitation from phytoplankton and self-shading, respectively. Due to seaweed treatment, average outflow concentration of NH₄ ⁺ was reduced from half of its pretreated level. This study showed that a weekly effluent supply at 2 kg m^?2 seaweed stocking density can sustainably support the growth of G. bailiniae as long as the dissolved nutrients are present at high levels.     

Supplementation effect(s) of organic acids and/or lipid to plant protein‐based diets on juvenile yellowtail,Seriola quinqueradiata Temminck et Schlegel 1845, growth and,nitrogen and phosphorus excretion

A feeding experiment was conducted to investigate the effect of organic acids and/or lipid supplementation on growth, utilization and environmental loading of nitrogen (N) and phosphorus (P) in juvenile yellowtail fed fishmeal (FM) and plant protein (PP) diets. Six diets as FM (FM‐based), FM+P (FM with inorganic P), FM+L (FM with lipid), PP+CA (PP with citric acid), PP+L+CA (PP with lipid and citric acid) and PP+L+FA (PP with formic acid) were formulated. Yellowtails were fed each of the diets in duplicate groups; once a day, 6 days a week to near satiation at water temperature 19.0–25.0 °C for 16 weeks. Fishmeal with inorganic P gave the best growth while PP+L+FA the lowest. However, growth increased in PP+CA and PP+L+CA. Addition of lipid significantly increased N and P retention resulting in significant reduction in N and P excretion. Citric acid and FA supplementation to PP diets also increased retention of P; hence, its excretion was lowered. Thus, CA, FA and lipid in juvenile yellowtail diets can help to partially replace FM with PP sources and reduce inorganic P use to minimize environmental loading from aquafeeds.     

Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)

We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.     

Histogenesis of neoformation in the endocrine pancreas of aging horses

Pancreatic tissue from 20 horses was examined using immunocytochemical techniques. In aged horses, neogenesis of endocrine cells, neoformation, and hyperplasia of islets occurred closely associated with the pancreatic duct; these changes were regarded as nesidioblastosis. In addition, pancreatic fibrosis accompanied by ductal proliferation and endocrine neogenesis was considered a regenerative change. Thus, the origin of neoformation in the endocrine pancreas was in the ductal system, and it is suggested that the pancreatic endocrine cells were of endodermal origin.     

Genetic relationship of litter traits between farrowing and weaning in Landrace and Large White pigs

We estimated genetic parameters in Landrace and Large White pig populations for litter traits at farrowing (total number born, number born alive, number stillborn, total litter weight at birth (LWB), and mean litter weight at birth) and those at weaning (litter size at weaning (LSW), total litter weight at weaning (LWW), mean litter weight at weaning (MWW), and survival rate from farrowing to weaning). We analyzed 65,579 records at farrowing and 6,306 at weaning for Landrace, and 52,557 and 5,360, respectively, for Large White. Single‐trait and two‐trait repeatability animal models were exploited to estimate heritability and genetic correlation respectively. Heritability estimates of LSW were 0.09 for Landrace and 0.08 for Large White. Genetic correlations of LSW with MWW were –0.43 for Landrace and –0.24 for Large White. Genetic correlations of LSW with LWW and LWB ranged from 0.5 to 0.6. The genetic correlation of MWW with LWW was positive, but that with LWB was negligible. The results indicate that utilizing LWW or LWB could improve LSW efficiently, despite the antagonistic genetic correlation between LSW and MWW.     

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.