Lolitrem B residue in fat tissues of cattle consuming endophyte-infected perennial ryegrass straw.

Lolitrems are neurotoxins found in endophyte-infected perennial ryegrass. Lolitrems, primarily lolitrem B, are the causative agents of ryegrass staggers in livestock. To guarantee the safety of meat produced from cattle consuming endophyte-infected perennial ryegrass, lolitrem B concentrations in tissues of Japanese Black cattle were determined using high-performance liquid chromatography. Lolitrem B was not detected in muscle, liver, kidney, or cerebrum of a Japanese Black cow with signs of ryegrass staggers. In contrast, perirenal fat contained 210 ppb lolitrem B. Three cows that received half as much perennial ryegrass straw as the cow with ryegrass staggers showed no clinical signs of ryegrass staggers. However, low concentrations of lolitrem B (less than 150 ppb) were detected in their fat tissue. These observations indicate that human exposure to the neurotoxic effect of lolitrem B through beef is unlikely. The amount of lolitrem B consumed by cattle can be estimated by the determination of lolitrem B in fat tissue.     

Detection and identification of the Candida species by 25S ribosomal DNA analysis in the urine of candidal cystitis

Candida species in clinical urine samples were identified directly by the newly developed method of PCR analysis on 25S ribosomal DNA (rDNA). Two dogs were referred to the Animal Medical Center, Nihon University School of Veterinary Medicine, Fujisawa, Kanagawa, Japan for the examination of chronic cystitis. Microscopic examination of urine samples from these dogs revealed yeast cells. Urine culture on Sabouraud's dextrose agar at 27 degrees C for 5 days produced white to cream colored colonies. The isolates were identifical to Candida albicans and C. parapsilosis by mycological examination, respectively. The nucleotide sequences of 25S ribosomal DNA from these urine isolates showed 99% similarity to those of a reference strain of Candida albicans or C. parapsilosis. The nucleotide sequences of 25S rDNA obtained directly from urine samples were also identical to C. albicans and C. parapsilosis, respectively. Confirming the results on the isolates cultured from the same urine samples. This PCR analysis method could be available for the direct identification of Candida species in urine samples within 2 days.     

Studies on the accumulation of nitrate in tomato fruit for canning

This study was carried out to investigate the factors concerning the accumulation of nitrate in tomato fruit and to find the preventive measures against heavy tin-dissolving in canned tomato juice caused by nitrate.The factors determining the accumulation of nitrate in the plant were as follows: the levels of nitrate, potassium, and calcium in the soil, the application of plant growth regulators, the tomato cultivar, and the light intensity. For obtaining fruits containing less than 3 ppm of nitrate-nitrogen, low nitrate cultivars should be grown in a sunny place on weak acid soil with split application of slow-acting nitrogen fertilizer, and a high calcium level. By proper treatment with plant growth regulators, nitrate-nitrogen content in the fruit can be decreased still more. It is also important that any particular growing-conditions in every tomato-growing district should be taken in consideration in the cultivation management for preventing the accumulation of nitrate-nitrogen.     

Protein synthesis and neurotrophin-dependent structural plasticity of single dendritic spines

Long-term potentiation (LTP) at glutamatergic synapses is considered to underlie learning and memory and is associated with the enlargement of dendritic spines. Because the consolidation of memory and LTP require protein synthesis, it is important to clarify how protein synthesis affects spine enlargement. In rat brain slices, the repetitive pairing of postsynaptic spikes and two-photon uncaging of glutamate at single spines (a spike-timing protocol) produced both immediate and gradual phases of spine enlargement in CA1 pyramidal neurons. The gradual enlargement was strongly dependent on protein synthesis and brain-derived neurotrophic factor (BDNF) action, often associated with spine twitching, and was induced specifically at the spines that were immediately enlarged by the synaptic stimulation. Thus, this spike-timing protocol is an efficient trigger for BDNF secretion and induces protein synthesis-dependent long-term enlargement at the level of single spines.     

Gustatory responses in Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel)

Gustatory neural responses of the Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel) to extracted compounds of prey organisms, such as amino acids, nucleotide‐related substances, organic acids and organic bases, were electrophysiologically recorded from the facial nerve supplying the anterior palate. Of the 17 amino acids tested, l ‐proline was the most potent amino acid at 10^?2 M, and its threshold was the lowest at around 10^?6 M. l ‐leucine, l ‐methionine, l ‐alanine, l ‐valine and l ‐isoleucine were also highly stimulatory at 10^?2 M; however, the other 11 amino acids examined were not as effective or were ineffective. Thus, the gustatory receptors for amino acids of the Pacific bluefin tuna show a narrowly tuned response profile. Among the seven nucleotide‐related substances tested, uridine‐5′‐monophosphate, inosine‐5′‐monophosphate and adenosine‐5′‐diphosphate were highly stimulatory, and their thresholds were 10^?4–10^?5 M. Inosine elicited a positive response at 10^?2 M but its response magnitude was not so high. Organic acids l ‐lactic and pyruvic acids were effective at 10^?2 M, but no response was elicited at 10^?3 M. Among organic bases, betaine was highly stimulatory, and its response magnitude at 10^?2 M is almost equal to that of l ‐proline at the same concentration. The threshold for betaine was determined to be at around 10^?5 M. Trimethylamine oxide and ammonium chloride were ineffective.     

Histopathological studies on viral nervous necrosis of sevenband grouper, Epinephelus septemfasciatus Thunberg, at the grow-out stage

Abstract Viral nervous necrosis caused by sevenband grouper nervous necrosis virus (SGNNV) has occurred in grow-out stages (0-3 years old) of sevenband grouper, Epinephelus septemfasciatus, since the 1980s. In the present study, based on histopathological features of the central nervous system (CNS) in naturally diseased fish, pernasal infection experiments using grow-out fish were performed and pernasal infection was established as a putative invasion route of SGNNV. The definite SGNNV-targeted cells were determined by histopathological studies including indirect fluorescent antibody test and electron microscopy. Nerve cells in the olfactory lobe were most extensively necrotized with vacuolation followed by infiltration of microglia and macrophages. Purkinje cells and Golgi cells were extensively infected in the cerebellum. Megalocells and small nerve cell nuclei were also infected in the preoptic area, thalamus, medulla oblongata and spinal cord. Only a few small nerve cells were infected in the olfactory bulb and optic tectum. The retina of some diseased fish displayed vacuolated bipolar cells of the inner nuclear layer and in the ganglion cell layer. These SGNNV-infected nerve cells displayed viroplasmic inclusions containing virions, vacuoles and myelin-like structures. Based on observed histopathological changes, the lesion of the CNS was characterized by encephalitis but not encephalopathy.     

Histopathological and haematological changes in amago salmon, Oncorhynchus rhodurus (Jordan & McGregor), fed a vitamin-D-free diet

Two purified diets supplemented with 0 or 20 000 IU vitamin D₃ kg^?1 of diet were fed to amago salmon, Oncorhynchus rhodurus (Jordan & McGregor), for 22 weeks. The feeding stopped when fish with deficiency signs including reduced growth, and caudal region and fin tip damage increased in number. Deficient fish had a thinned epidermis consisting of atrophied and necrotic epidermal cells over the basal cells. In lesions of the caudal peduncle, the skin and the underlying musculature were extensively necrotized. The liver showed degenerated and necrotic hepatocytes. Respiratory epithelium and cardiac muscle fibres were swollen. Hypocalcaemia was evident. No pathological changes were found in bone, eyes, brain and alimentary tract. The control groups fed a nutritionally complete diet grew well and displayed no pathological changes. The pathological changes were reversed after 4 weeks of feeding of vitamin-D₃-suppliemented diet.     

Genetic analysis of phosphoglucoisomerase isozymes in cultivated and wild species of section Cepa in Allium

Phosphoglucoisomerase (PGI) was studied to establish the genetic system of the isozymes in section Cepa of Allium. The results of isozyme analyses using 15 cultivars and clones in two cultivated species A. fistulosum and A. cepa, eight strains in four wild species A. altaicum, A. galanthum, A. oschaninii and A. vavilovii and a total of 10 hybrids between A. fistulosum and A. cepa and between the cultivated and wild species revealed that: 1) PGI was a dimeric enzyme; 2) the two cultivated and four wild species had a common gene locus Pgi-1 where six alleles were identified; and 3) A. fistulosum had Pgi-1 ^F-1 and Pgi-1 ^F-2, A. cepa had Pgi-1 ^A-1 and Pgi-1 ^A-2, A. altaicum had Pgi-1 ^F-1, A. galanthum and A. vavilovii had Pgi-1 ^A-1, and A. oschaninii had Pgi-1 ^A-1, Pgi-1^O-1 and Pgi-1 ^O-1. The intra- and inter-specific polymorphisms of PGI isozymes are expected to be valuable tools for the genetics and practical utilization of the wild species in section Cepa.     

Genetic diversity of group A rotaviruses associated with repeated outbreaks of diarrhea in a farrow-to-finish farm: identification of a porcine rotavirus strain bearing a novel VP7 genotype, G26

ABSTRACT: Group A rotaviruses (GARs) are one of the most common causes of diarrhea in suckling pigs. Although a number of G and P genotypes have been identified in porcine GARs, few attempts have been made to study the molecular epidemiology of these viruses associated with diarrhea outbreaks within a farm over an extended period of time. Here, we investigated the molecular characteristics of GARs that caused four outbreaks of diarrhea among suckling pigs in a farrow-to-finish farm over the course of a year. G and P genotyping of GARs detected at each outbreak demonstrated genetic diversity in this farm as follows: G9P[23] was detected at the first outbreak, G9P[13]/[22] and G9P[23] at the second, G3P[7] at the third, and G9P[23], G5P[13]/[22], and P[7] combined with an untypeable G genotype at the fourth. Sequence analysis of the detected GARs revealed that such genetic diversity could have resulted not only from the introduction of new GAR strains, but also from gene reassortment between GAR strains within the farm. Further, the GAR strain carrying the untypeable G genotype was shown to be a novel porcine GAR bearing a new G26 genotype, as confirmed by the Rotavirus Classification Working Group.     

Characterisation of the carboxylesterase enzyme cauxin in the seminal fluid of the cat

The carboxylesterase cauxin is a major urinary protein in cats that is also found in seminal fluid (SF). This study investigated cauxin in feline SF including biochemical features, concentration, distribution and gene expression in epididymal tissue, and its reaction with acylglycerol substrates.Monomeric, dimeric, and/or multimeric forms of cauxin carrying N-glycosylations were detected on Western blots of feline SF but most were monomeric. Cauxin concentrations were markedly lower in SF (0.042 ± 0.020 mg/mL) than in urine (∼0.5 mg/mL) and cauxin gene expression was 60-fold lower in the epididymis than in the kidney. Immunohistochemical examination localised cauxin within the stereocilia and cytoplasm of epithelial cells lining the caput and corpus epididymis. Cauxin-positive spermatozoa were detected in the lumen of the cauda epididymis but not in the cytoplasm of the epithelial cell lining. Using an in vitro assay, cauxin hydrolysed saturated 1-mono- but not di- and tri-acylglycerols. The results suggest that cauxin secreted from the caput and corpus epididymis acts as an esterase on lipid within feline SF.