Expression analysis of the type I keratin protein keratin 33A in goat coat hair

The coat of a goat, like that of many mammalian species, consists of an outer coat of coarse hairs and an under coat of fine, downy hairs. The coarse guard hairs are produced by primary follicles and the finer cashmere hairs by secondary follicles. We previously reported that hair keratins are components of cashmere hair, and proteomic analysis revealed that their expression is elevated in winter coat hair. To determine detailed characterization, we have cloned keratin 33A gene, a major highly expressed keratin in winter, and then analyzed the expression of goat hair coat. By Western analysis, we detected that keratin 33A protein is expressed only in hair coat among the various goat tissues. Moreover, the expression level in winter has increased in cashmere high‐producing Korean native breed, whereas the expression levels between summer and winter had not changed in cashmere low‐producing Saanen. In addition, by immunohistochemistry we determined that keratin 33A is localized in the major cortex portion of cashmere fiber. These results confirm that keratin 33A is a structural protein of goat cashmere hair fiber.     

Epidemiological survey,general blood biochemistry,and histological examination of slaughtered heavy horse breeds with hemorrhage in the adipose tissue in the crest of the neck

Fifty-four slaughtered horses were classified into groups having adipose tissue in the crest of the neck with or without hemorrhage (AH and NH groups, respectively). Blood biochemical tests (Alb, TP, T-bil, GOT, GPT, LDH, T-cho, and BUN) and an epidemiological survey (age, gender, weight, origin, breed, BCS, CNS, and hoof disease) were performed. T-bil tended to be high, while the other parameters were normal. Weight, BCS, and CNS were higher in the AH group (P<0.05). GOT was lower in the AH group (P<0.05). It was suspected that the horses in the AH group had lipomatosis. It was assumed that the adipose tissue of the horses in the AH group contained damaged capillaries, and inflammation was confirmed based on evidence of macrophages and lymphocytes.     

Characteristics of cytochrome P450-dependent metabolism in the liver of the wild raccoon,Procyon lotor

Wildlife is exposed to a wide range of xenobiotics in the natural environment. In order to appropriately assess xenobiotic-induced toxicity in wildlife, it is necessary to understand metabolic capacities. Carnivores, in general, have low metabolic abilities, making them vulnerable to a variety of chemicals. Raccoons (Procyon lotor) in the wild have been found to have high levels of xenobiotics. However, little is known about the metabolic capacity of the cytochrome P450 (CYP) enzymes in this species. Thus, this study used liver samples to investigate the characteristics of CYP enzymes in wild raccoons. In 22 wild raccoons, CYP concentrations in hepatic microsomes were examined. To better understand the properties of CYP-dependent metabolism, in vitro metabolic activity studies were performed using ethoxyresorufin, pentoxyresorufin and testosterone as substrates. In addition, three raccoons were fed commercial dog food in the laboratory for one week, and the effects on CYP-dependent metabolism were investigated. In comparison to other mammalian species, raccoons had very low concentrations of CYP in their livers. In an in vitro enzymatic analysis, raccoons’ ethoxyresorufin O-deethylase (EROD) and pentoxyresorufin O-depentylase (PROD) metabolic capacities were less than one-fifth and one-tenth of rats’, respectively. These results indicate the possible high risk in raccoons if exposed to high levels of environmental xenobiotics because of their poor CYP activity. In this study, the features of CYP-dependent metabolism in wild raccoons are described for the first time.     

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.     

N-Acetyl-d-Glucosamine-Binding Lectin in Acropora tenuis Attracts Specific Symbiodiniaceae Cell Culture Strains

Many corals establish symbiosis with Symbiodiniaceae cells from surrounding environments, but very few Symbiodiniaceae cells exist in the water column. Given that the N-acetyl-d-glucosamine-binding lectin ActL attracts Symbiodiniaceae cells, we hypothesized that corals must attract Symbiodiniaceae cells using ActL to acquire them. Anti-ActL antibody inhibited acquisition of Symbiodiniaceae cells, and rearing seawater for juvenile Acropora tenuis contained ActL, suggesting that juvenile A. tenuis discharge ActL to attract these cells. Among eight Symbiodiniaceae cultured strains, ActL attracted NBRC102920 (Symbiodinium tridacnidorum) most strongly followed by CS-161 (Symbiodinium tridacnidorum), CCMP2556 (Durusdinium trenchii), and CCMP1633 (Breviolum sp.); however, it did not attract GTP-A6-Sy (Symbiodinium natans), CCMP421 (Effrenium voratum), FKM0207 (Fugacium sp.), and CS-156 (Fugacium sp.). Juvenile polyps of A. tenuis acquired limited Symbiodiniaceae cell strains, and the number of acquired Symbiodiniaceae cells in a polyp also differed from each other. The number of Symbiodiniaceae cells acquired by juvenile polyps of A. tenuis was correlated with the ActL chemotactic activity. Thus, ActL could be used to attract select Symbiodiniaceae cells and help Symbiodiniaceae cell acquisition in juvenile polyps of A. tenuis, facilitating establishment of symbiosis between A. tenuis and Symbiodiniaceae cells.     

Partial cDNA sequence of a relaxin‐like factor (RLF) receptor,LGR8 and possible existence of the RLF ligand‐receptor system in goat testes

Relaxin‐like factor (RLF), also known as insulin‐like factor 3 (INSL3), is produced by testicular Leydig cells, but its specific receptor LGR8 (leucine‐rich repeat family of G‐protein‐coupled receptor 8) has not been identified in goats. This study aimed to identify complementary DNA (cDNA) sequences of goat LGR8, and characterize the expression of both RLF and LGR8 in goat testes by RT‐PCR and immunohistochemistry. Testes were collected from immature (3‐month‐old) and mature (24‐month‐old) Saanen goats, and partial cDNA sequences of the goat homologue of human LGR8 were identified. The sequence encoded a reduced peptide sequence of 167 amino acids, which corresponded to transmembrane regions 2 through 5, followed by the beginning of intracellular loop 3 of human LGR8. Expression of both LGR8 and RLF genes was drastically increased in mature testes compared with immature ones. Although RLF protein was restricted to Leydig cells, LGR8 protein was detected in both Leydig cells and seminiferous epithelial cells (possibly germ cells and Sertoli cells). These results reveal a possible existence of the RLF‐LGR8 ligand‐receptor system within the goat testis, suggesting that RLF may play a role in testicular function through LGR8 on Leydig cells and seminiferous epithelial cells in an autocrine and/or paracrine manner.     

A novel blast resistance locus in a rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivar,Chumroo, of Bhutan

The rice cultivar ‘Chumroo’ is commonly cultivated in the mid- and high-altitude areas of Bhutan. This cultivar has shown durable blast resistance in that area, without evidence of breakdown, for over 20 years. Chumroo was inoculated with 22 blast isolates selected from the race differential standard set of Japan. The cultivar showed resistance to all the isolates. To identify the resistance gene(s), Chumroo was crossed with a susceptible rice cultivar, Koshihikari. The F₁ plants of the cross showed resistance. Segregation analyses of 300 F₃ family lines fitted the segregation ratio of 1:2:1 and indicated that a single dominant gene controls the resistance to a blast isolate Ao 92-06-2 (race 337.1). The Chumroo resistance locus (termed Pi46(t)) was mapped between two SSR markers, RM6748 and RM5473, on the terminal region of the long arm of chromosome 4, using linkage analysis with SSR markers. The nearest marker, RM5473, was linked to the putative resistance locus at a map distance of 3.2 cM. At the chromosomal region, no true resistance genes were identified, whereas two field resistance genes were present. Therefore, we designated Pi46(t) as a novel blast resistance locus.