Demonstration of bovine herpesvirus type 1 and Mannheimia haemolytica antigens in specimens stored for up to 22 months in buffered formalin

Bovine herpesvirus type 1 (BHV-1) and Mannheimia haemolytica antigens were demonstrated in lung tissues that were stored in 10% neutral phosphate buffered formalin for 1 to 22 months using the immunoperoxidase method. There were no differences observed in terms of labelling intensity and distribution of M. haemolytica antigens between specimens stored for 1 and 22 months. The labeling intensity in sections from 2-cm thick specimens was comparable to those from 0.2-cm thick specimens. There was no difference observed between pronase-treated and -untreated sections. However, for BHV-1, the labeling intensity in untreated sections was reduced in tissues that had been stored from 12 to 22 months. Sections from thin specimens stored in neutral buffered formalin for 22 months exhibited a stronger staining intensity than those from thick specimens.     

A laboratory-based assessment of phosphorus and nitrogen loading from currently available commercial carp feeds

ABSTRACT: To clarify the total phosphorus (P) and nitrogen (N) loading from carp culture, five commercial diets were selected from a major location in Japan, Lake Kasumigaura, for a 12-week feeding trial. These diets were prepared as per the 'Kasumigaura Feed Standard' (crude protein < 35% and digestible energy > 3.5kcal/g), and total P ranged from 1.4% to 2.0%. However, for most of the diets the P available was lower than the requirement level. A control diet was formulated with 25% fishmeal to comply with that standard and contain adequate available P. Duplicate groups of juvenile carp were fed the aforementioned diets to satiation, three times a day, six days a week throughout the trial. Growth performance was significantly higher for the control group and values of P absorption (20.4–47.0%) and retention (14.0–36.3%) varied widely among the groups. Consequently, the total P loading (kg/t production) values based on retention fluctuated from 14.8 to 26.4 among the commercial diet groups compared with the low level of 8.5 for the control group. Similarly, the total N loading (kg/t production) values varied from 30.9 to 86.0 and was lowest for the control group. A higher whole body lipid and lower bone P and Ca confirmed the deficiency of the dietary available P in commercial diets. Better growth and comparatively less P and N loading rates were observed in the diet that had sufficient available P, not to mention that the control diet ranked best. It was concluded that an inadequacy of available P among the commercial diets affects the growth of carp and produces high P and N loading into the water. Therefore, if the commercial diets do not supply adequate levels of available P to carp, growth is negatively affected and may result in greater waste loading.     

Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method

ABSTRACT: Protoplast fusion between different species of Streptomyces was performed using a liquid regeneration method developed for a rapid and simple preparation of the fusants. Consequently, new clones, which could not be obtained using the conventional agar regeneration method, were obtained. In the crosses between S. griseus and S. durhamensis , and between S. californicus and S. catenulae , eight and two recombinants, respectively, were obtained using the liquid regeneration method. Conversely, in the case of crosses between S. ornatus and S. catenulae , and between S. ornatus and S. vendargensis , seven recombinants each were obtained using only the agar method. The physiological characteristics, such as the assimilation of carbohydrate and antibiotic resistance, of these fusants differed considerably from those of their parental strains. Using the proposed liquid regeneration method, a simpler and quicker procedure for protoplast fusion is described.     

4-Nitro-3-phenylphenol has both androgenic and anti-androgenic-like effects in rats

To investigate the effect of endocrine disruption of 4-nitro-3-phenylphenol (PNMPP) on immature male Wistar-Imamichi rats, the rat pituitary was exposed to PNMPP (10^–5–10^–9 M) for 24 h with or without gonadotropin-releasing hormone (GnRH) in experiment I. In addition, the Leydig cells (10^–5–10^–9 M) were exposed to PNMPP for 24 h with or without human chronic gonadotropin (hCG) in experiment II. Our results showed that the PNMPP at 10^–5–10^–7 M suppressed follicle-stimulating hormone (FSH) and luteinizing hormone (LH) productions from GnRH-stimulated pituitary cells. At the same time, PNMPP 10^–5–10^–7 M induced an increase in testosterone production from the Leydig cells treated with or without hCG. Based on our results, it can be concluded that that PNMPP might have both androgen agonist action by decreasing FSH and LH production in the pituitary and anti-androgenic action by increasing testosterone production in the Leydig cell.     

Experimental approach to prezygotic chromosome screening using only a single pair of gametes in mice

During in vitro embryo production, chromosome screening is essential to prevent pregnancy losses caused by embryonic chromosome aberrations. When the chromosome screening is completed before fertilization, gametes are effectively utilized as genetic resources. The aim of this study was to investigate whether chromosome screening of gametes accompanied by fertilization would be feasible using a single mouse spermatozoon and oocyte. Metaphase II oocytes were divided into a cytoplast and a karyoplast. For genome cloning of the gametes, androgenic and gynogenic embryos were produced by microinjection of sperm into cytoplasts and parthenogenetic activation of karyoplasts, respectively. Pairs of blastomeres from androgenic and gynogenic embryos were fused electrically to produce diploid embryos, which were transferred into pseudopregnant surrogate mothers to examine fetal development. Blastomeres from androgenic and gynogenic embryos were individually treated with calyculin A—a specific inhibitor of type 1 and 2A protein phosphatases—for 2 h to induce premature chromosome condensation. Thereafter, chromosome analysis of blastomeres, reflecting the genetic constitution of individual spermatozoa and oocytes, was performed, and we confirmed that most of the androgenic and gynogenic 2-cell embryos had a haploid set of chromosomes in their sister blastomeres. The reconstructed embryos from blastomeres of androgenic and gynogenic 2-cell embryos could be implanted and develop into live fetuses, albeit at low efficiency. This study indicates that prezygotic chromosome screening and embryo production using a single pair of gametes may be practicable.