Molecular cloning, chromosomal location, and biological activity of porcine interleukin-21

A pig interleukin-21 (IL-21) cDNA was successfully cloned and sequenced from porcine peripheral blood lymphocytes (PBL) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin P (PHA), 50 ng/ml phorbol 12-myristate 13-acetate (PMA), and 0.5 microg/ml anti-porcine CD3 antibody for 48 hr. The open reading frame of the porcine IL-21 cDNA is 459 base pairs in length and encodes 152 amino acids. The predicted amino acid sequence of the porcine IL-21 shows 86.2%, 77.7%, and 58.4% identity to the bovine, human, and murine IL-21, respectively. The porcine IL-21 gene was mapped to porcine chromosome 8 (8q22-->q23) by means of fluorescence in situ hybridization and radiation hybrid mapping, where the porcine IL-2 gene had been mapped nearby. The recombinant porcine mature IL-21 expressed by E. coli induced dose-dependent proliferation and IFN-gamma production from a human NK cell line, NK0. The porcine IL-21 identified in this study will be helpful for the enhancement of innate immune responses of pigs.     

Polymorphisms in the canine glutamate transporter-1 gene: identification and variation among five dog breeds

Excitatory amino acid transporters (EAATs) are important for terminating glutamatergic neurotransmission and protect central nervous system (CNS) neurons from glutamatergic excitotoxicity. We selected these genes as targets that may relate to canine behavioral traits. After screening four EAAT genes (glutamate transporter-1; GLT-1, excitatory amino acid transporter 4; EAAT4, excitatory amino acid carrier; EAAC1, glutamate/aspartate transporter; GLAST) for single nucleotide polymorphisms (SNPs), we identified two silent SNPs (C129T and T471C) in the GLT-1 gene. We genotyped 193 dogs of 5 breeds and found significant variation among breeds in these two SNPs in GLT-1. The C129T polymorphism was not observed in Malteses and Miniature Schnauzers. These results suggest that polymorphisms in the GLT-1 gene may be useful markers for examining how the genetic background relates to the behavioral traits of dogs.     

A comparison of the behavioral profiles of purebred dogs in Japan to profiles of those in the United States and the United Kingdom

To clarify the behavioral profiles of 56 pure breeds of dogs in Japan, 96 small-animal veterinarians participated in a questionnaire survey using the same criteria as in preceding studies conducted in the United States and the United Kingdom. We found significant differences among breeds in all behavioral traits examined. In addition, gender differences were revealed in terms of aggression to dogs, territorial defense, excitability, general activity, dominance over owner, destructiveness, watchdog barking, and snapping at children, which were all rated higher in males than females, whereas obedience training and housebreaking ease were rated higher in females. No gender differences were evident in playfulness, excessive barking, or affection demand. Using factor analyses, "aggressiveness", "reactivity", and "trainability" were determined to be consistent with results found in the US and UK surveys. On the basis of these factor scores, seven groups of breeds were determined by cluster analysis to compare to the US survey; 22 of the 38 breeds common with the US survey were categorized into the same groups as those in that survey. The results demonstrated differences in canine behavioral predisposition among breeds and between genders. The similarity in the results between our study and previous surveys, which involved distinct geographical locales, suggests that the genetic basis of breed-specific temperamental traits is manifested irrespective of the cultural or regional identities of the owners.     

The oral administration of thermophile-fermented compost extract and its influence on stillbirths and growth rate of pre-weaning piglets

Food produced via fermentation with mesophilic bacteria has been used to confer health benefits. In contrast, mammalian physiological responses to the intake of thermophile-fermented products have not been thoroughly investigated. We examined the effects of administering a compost extract consisting of fermented marine animals with thermophiles, including Bacillaceae, to pregnant sows and piglets. Retrospective studies were performed on two different swine farms (n=330-1050 sows). The rate of stillbirth was markedly lower in all parities of the compost extract-fed group compared to those of the control group (p≦0.001). Additionally, the birth to weaning period of newborns was significantly shorter (p<0.0001), while the ratio of weanlings per liveborn piglets was increased by more than 6.5% in the compost extract-fed group. Thus thermophiles and their products in the compost extract might promote growth and reduce stillbirths of piglets during the birth to weaning period.     

Cytologically atypical anal sac adenocarcinoma in a dog

A 10-year-old intact female Shetland Sheepdog with tenesmus had a subcutaneous mass at the left ventral aspect of the anus. On cytologic examination, 2 types of cells were observed. Most of the cells were oval to polygonal and had elliptical or elongate nuclei and a moderate amount of pale to basophilic cytoplasm. The remaining cells had round to oval nuclei and pale to basophilic cytoplasm. Cells of both types were loosely adhered to each other and were arranged in rosette-like structures. Both neoplastic cell types had fine homogenous chromatin and either a small indistinct nucleolus or no visible nucleolus. Mild anisokaryosis and anisocytosis were observed. Histologically, the mass consists of glandular structures formed by cuboidal cells admixed with bundles of spindle cells. Eosinophilic PAS- and Alcian blue-positive secretory material was found in the center of some glandular structures. Both neoplastic cell types had positive staining with paradoxical concanavalin A and expressed cytokeratin, but not vimentin, S-100, α-smooth muscle actin, or desmin. Based on location and histologic and immunohistochemical features, the final diagnosis was adenocarcinoma of the apocrine gland of the anal sac, which should be included as a cytologic differential diagnosis when spindle cells and typical epithelial cells are observed in masses in the region of the anal sac of dogs.     

Purification of N-acetyllactosamine-binding activity from the porcine sperm membrane: possible involvement of an ADAM complex in the carbohydrate-binding activity of sperm

Although the importance of carbohydrate recognition by sperm during egg zona pellucida binding has been widely reported, the sperm molecular species that recognize the carbohydrates are poorly characterized. Our previous cytochemical study indicated that two kinds of carbohydrate-binding proteins are expressed on porcine sperm heads-one recognizes N-acetyllactosamine (Galβ1-4GlcNAc-), and the other recognizes the Lewis X structure (Galβ1-4(Fucα1-3)GlcNAc-). For this report, we used proteomic techniques to characterize the sperm proteins that bind N-acetyllactosamine. Porcine sperm plasma membrane was solubilized with a detergent solution and subjected to sequential chromatography with dextran sulfate agarose, affinity, and hydroxyapatite, and the binding activities in the eluates were monitored by a solid-phase binding assay. The tryptic peptides of two proteins most likely associated with the binding activities were subjected to tandem mass spectrometry sequencing. A subsequent database search identified one of the two proteins as predicted disintegrin and metalloprotease domain-containing protein 20-like (XP_003128672). The other protein was identified as disintegrin and metalloprotease domain-containing protein 5 (AB613817) by database searches for homologous amino acid sequences, cDNA cloning, nucleotide sequencing and nucleotide database searches. Furthermore, two-dimensional blue native/SDS-PAGE demonstrated that they formed a variety of non-covalent complexes. Therefore, these ADAM complexes probably are responsible for the N-acetyllactosamine-binding activity. An affinity-purified fraction containing these ADAM complexes showed zona pellucida-binding activity, though the activity was relatively weak, and the presence of another zona pellucida-binding protein that probably works in concert with these ADAM complexes was suggested. Immunofluorescence testing suggested that ADAM20-like was localized on the anterior part of the sperm plasma membrane.     

Mapping of quantitative trait loci controlling seedling vigor in rice (Oryza sativa L.) under submergence

Submergence-induced suppression of seedling vigor is a serious constraint particularly in the direct seeding rice cultivation system. To identify quantitative trait loci (QTLs) associated with seedling vigor in rice under submergence, a mapping population of 98 Backcross Inbred Lines derived from a cross of Nipponbare/Kasalath//Nipponbare was used. Phenotypic evaluation of seedling vigor under submergence was based on shoot length (SHL), root length (RTL) and shoot fresh weight (SFW) using a test tube bioassay method. Thirty-two putative QTLs were detected among which 7 were for SHL, 11 for RTL and 14 for SFW. Phenotypic evaluation was also made of the parental lines and a set of 54 chromosome segment substitution lines in which Nipponbare segments were substituted for by their homologous Kasalath segments covering the entire rice genome. Two QTLs with more than 10 % contribution to the total phenotypic variance were verified for SHL, and at least one for RTL and six for SFW on chromosomes 1, 3, 4, 6 and 7 at the 1 % significance level. Among these, all but two showed reductions in one, two or all three traits. Our present and previous results suggest that the Nipponbare genome has a potential to improve seedling vigor under submergence and that japonica germplasms can be used to breed for this important trait in indica rice.     

Basilaphelenchus gorganensis n. sp. (Aphelenchoidea,Tylaphelenchinae) from wood from northern Iran

Basilaphelenchus gorganensis n. sp. is described and illustrated from wood and bark of a dead tree from northern Iran. The new species is characterized by female body length (415–559 µm), three‐lined lateral fields, a sclerotized cephalic vestibule and cephalic framework, thin stylet with three elongate backwardly directed knobs, small spherical to oval metacorpus, with small and posteriorly located valve, simple vulva without any flap apparatus, 59‐ to 79‐µm‐long post‐vulval uterine sac, functional rectum and anus and dorsally convex, ventrally concave, usually ventrally bent conical female tail with a sharp tip. Males are common, apparently functional and characterized by well‐curved spicules, three pairs of small caudal papillae and no bursa at tail tip. Molecular phylogenetic inferences using partial sequences of small and large subunit ribosomal RNA genes (SSU and LSU rDNA) from different isolates of the new species revealed it differs from currently sequenced species and belongs to the Tylaphelenchinae clade.     

Turbid water induces refuge behaviour of a commercially important ayu: A field experiment for interstream movement using multiple artificial streams

Riverine systems are inherently dendritic in nature, and turbid water may not necessarily flow equally throughout an entire catchment. Tributaries have the potential to serve as refugia from temporarily turbid water in the main stem. As ayu (Plecoglossus altivelis) are one of the important species for the inland fisheries industry and recreational fishing, their behaviour in a branched river network is a primary concern. We released ayu with radio‐transmitting tags into two experimental streams that joined at a downstream pool. Turbid water was released into one of the streams, and we identified whether avoidance movement of ayu between the two experimental streams was triggered by flowing turbid water. The suspended solid concentration elevated rapidly after adding the turbid water and was maintained at more than 400 mg/L for 3 hr. Avoidance movement of ayu was triggered when the suspended solid concentration exceeded approximately 200 mg/L. Then, most ayu moved from the turbid stream to the nonturbid one, leading to a difference in ayu density between the two streams. Therefore, we demonstrated that turbid water induced interstream movement of a commercially important ayu as refuge behaviour, suggesting that river connectivity is important for the persistence of the ayu.