Effect of temperature, age class, and growth on induction of aestivation in Japanese sandeel (Ammodytes personatus) in Ise Bay, central Japan

We investigated the effects of temperature, age class, and growth on induction of aestivation in the Japanese sandeel (Ammodytes personatus) by means of rearing experiments and field surveys in Ise Bay, central Japan. Acoustic surveys and sampling of shoaling and aestivating sandeels indicated that the sandeels completed aestivation when the water temperature increased to 17–20°C. A rearing experiment showed that the mortality of sandeels reared without sand was greater than it was with sand, under replete feeding conditions. In a non‐aestivation rearing experiment without sand, the condition factor of the sandeels decreased when water temperatures exceeded 20°C, although they were fed to repletion. These results suggest that aestivation is induced by rising seawater temperature and plays an important role in energy conservation. The age‐1 group tended to start aestivation earlier than the age‐0 group in the field. Within the age‐0 group, larger sandeels tended to enter aestivation earlier. These observations indicate that fat reserves are an important cue to start aestivation.     

Alanine racemase activity in the microalga <Emphasis Type="Italic">thalassiosira</Emphasis> sp.

ABSTRACT:   In this paper, the authors report the detection of alanine racemase activity in the marine diatom Thalassiosira sp. Since the Thalassiosira sp. was cultured under germ-free conditions, it appeared that D-alanine was not derived from bacteria but was produced through catalysis by algal alanine racemase. The rate of conversion of L-alanine to D-alanine was approximately the same as that for the reverse reaction, and the enzyme catalyzed the equilibration of the D- and L-forms. The crude enzyme preparation obtained from the cells at the stationary phase of the growth cycle had an optimal pH of approximately 9.5. The Lineweaver–Burk analysis showed that the K _m for D- and L-alanine was 16.5 mM and 29.4 mM, respectively. It appears that the enzyme is highly specific for D- or L-alanine because it does not catalyze the racemization of other amino acids. In addition, after gel filtration, the enzyme did not require exogenous pyridoxal 5'-phosphate (PLP) for its activity, however, the effects of several chemicals suggest that the enzyme may be PLP-dependent. The enzyme is more similar to that found in invertebrates when compared with that found in bacteria. This is the first report on the occurrence of alanine racemase activity in the microalga Thalassiosira sp.     

A Case Report of a Malignant Fibrous Histiocytoma in a T-cell Receptor β Chain and p53 Double-knockout Mouse

A subcutaneous tumor was found in the right abdomen of a 16-week-old male TCRβ and p53 double-knockout mouse. The tumor had indistinct borders with the surrounding tissue. The cut surface after formalin fixation was pale yellowish white, partially dark red and partly white. Histologically, the tumor was composed of three distinct regions. The first region showed pleomorphic cells arranged in sheets. The second region showed spindle cells arranged in interlacing fascicles. The final region contained a mixture of the above mentioned two types of cells. Furthermore, a small amount of collagen fibers, round cells, multinucleated giant cells, and cells with eosinophilic granules were observed between these tumor cells. Immunohistochemical examination and electron microscopy identified that the pleomorphic cells and spindle cells were histiocytes and fibroblasts, respectively, and that the round cells were undifferentiated mesenchymal cells. Based on these findings, the tumor was diagnosed as a malignant fibrous histiocytoma.     

Practical aspects of the fertility of dairy cattle

To establish sustainability in the dairy industry, it is important that cows become pregnant at a biologically optimal time and at an economically profitable interval after calving. In this review, the results obtained from Holstein cattle in an experimental herd for dairy research are summarized. First, the effect of age at first calving of heifers on productive and reproductive performance was examined. A reduction in calving age from 25.1 to 21.5 months with the same growth rate during the first 12 months after birth had no negative effects on the heifers' performance. Second, the postpartum follicular dynamics of lactating cows were traced in relation to their fertility, and the emergence and fate of cystic ovarian follicles were examined. The premature initiation of ovarian activity does not always improve the fertility of cows as indicated by the number of days open. Third, the occurrences of anestrous ovulation during the early postpartum period were analyzed with reference to the frequency of reversion to anestrus. The premature onset of estrous activity also did not improve fertility, and relapse back into anestrus after the onset of the estrous cycle often occurred during the breeding period. Fourth, some indices for the occurrence of postpartum reproductive events were evaluated as an indicator of the reproductive performance of lactating cows. The milk yield and percentage of body weight loss could be indicators for reproductive events. Finally, the potency of a pedometry system for the detection of typical and atypical estrous behaviors of heifers and lactating cows was evaluated in terms of efficiency and accuracy. The location of the pedometers and housing conditions for the animals affected the estrus detection of the system. These results represent the reproductive potential of modern high-yielding dairy cattle and provide a baseline to evaluate their reproduction.     

Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP)

We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.     

Mycobacterium ulcerans infection in an Indian flap-shelled turtle (Lissemys punctata punctata)

We report an atypical mycobacterial infection in an Indian flap-shelled turtle, Lissemys punctata punctata, that died in an aquarium in Japan. At necropsy, the turtle showed multiple white nodules on the capsular surface and parenchyma of various organs such as the liver, spleen, intestine, and lung. Histologically, granulomatous inflammation surrounding a central zone of necrosis was observed. Sections stained by the Ziehl-Neelsen method revealed numerous acid-fast bacilli in the cytoplasm of macrophages and in the central area of necrosis. The organisms were identified as a mycobacterial species by PCR and nucleotide sequence analysis and revealed 98-100% homology to M. ulcerans. This is, to our knowledge, the first report of mycobacteriosis due to M. ulcerans in a turtle.     

Reduction of urinary nitrogen excretion and ammonia emission from slurry by feeding a low protein diet supplemented with apple pomace to growing pigs

An experiment was conducted to determine the effects of supplementing a reduced crude protein (CP) diet with apple pomace on the urinary nitrogen excretion and ammonia emission from slurry in the growing pigs. Four barrows (35 kg bodyweight) were assigned to one of two diets. Each pig was placed in a metabolism cage and fed a low CP, amino acid-supplemented diet (CP 11.33%; low CP diet) or a low CP, amino acid-supplemented diet containing 23.08% dried apple pomace (CP 9.47%; apple pomace diet) for two 14-day experimental periods. After the completion of the first period, the pigs were switched to the other diet. Urine and feces were quantitatively collected daily for 5 days after a 9-day adaptation period. The daily nitrogen intake for the low CP diet and the apple pomace diet was 17.76 g/pig and 18.64 g/pig, respectively. The pigs fed the apple pomace diet excreted more fecal nitrogen (6.86 g/day) than the pigs fed the low CP diet (3.63 g/day) ( P  < 0.001), but urinary nitrogen excretion with the apple pomace diet was 3.11 g/day, which was much lower than that for the low CP diet (5.95 g/day) ( P  < 0.001). The daily ammonia emission from the mixture of urine and feces determined by an in vitro method was much lower for pigs fed the apple pomace diet (120 mg) than that for pigs fed the low CP diet (603 mg) ( P  < 0.01). The addition of apple pomace to a reduced CP, amino acid-supplemented diet reduces urinary nitrogen excretion and thereby ammonia emission.     

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.     

Characterization of ammonia-assimilating bacteria in a lagoon for wastewater from a paddock of dairy cattle

We investigated microorganisms that assimilated ammonia in lagoon treatment processes. Ammonia‐assimilating microorganisms were detected by nitrogen‐limited medium that contained ammonia as the sole nitrogen source. Numbers of ammonia‐assimilating aerobes (log CFU/g) were 3.4, 4.8, 5.0, 4.8 and 5.0 (log CFU/mL) on the culture plate incubated at 4°C, 10°C, 15°C, 20°C and 25°C, respectively. Many isolates used ammonia in high rates when they were purely cultivated in nitrogen‐limited medium added to sterilized lagoon extract. Many of them used ammonia even when they were cultivated in media containing viable microbial flora of the lagoon. Among them, enterobacteriaceae and Pseudomonas sp. were identified by analysis of 16S ribosomal DNA.     

Expression of monocarboxylate transporter 1 (MCT1) in the dog intestine

In this study, the expression and distribution of monocarboxyolate transporter 1 (MCT1) along the intestines (duodenum, jejunum, ileum, cecum, colon and rectum) of dogs were investigated at both the mRNA and protein levels. The expression of MCT1 protein and its distribution were confirmed by Western blotting and immunohistochemical staining using the antibody for MCT1. We identified mRNA coding for MCT1 and a 43-kDa band of MCT1 protein in all regions from the duodenum to the rectum. Immunoreactive staining for MCT1 was also observed in epithelial cells throughout the intestines. MCT1 immunoreactivity was greater in the large intestine than in the small intestine. MCT1 protein was predominantly expressed on the basolateral membranes along intestinal epithelial cells, suggesting that MCT1 may play an important role in lactate efflux and transport of short-chain fatty acids (SCFAs) to the bloodstream across the basolateral membranes of the dog intestine.