Genetic diversity of Hungarian canine distemper virus strains

To achieve proper diagnosis of dogs based on acute clinical symptoms and poorly preserved field samples taken from animals that died due to canine distemper (CD), a new differential diagnostic test has been developed based on polymerase chain reaction (PCR). In this study, more than 150 samples collected from dogs showing respiratory, gastrointestinal and neurological signs suggesting canine distemper virus (CDV) infection were examined. The samples consisted of urine, blood and nasal swabs collected from clinically ill patients, sent to our laboratory by clinicians from various veterinary clinics throughout Hungary. Various organs collected during the necropsy of dogs with pathological changes that suggested CDV infection were also included. Three distinct PCRs were designed. For diagnostic purposes, a primer pair specific to a 409 bases-long segment within the conservative part of the large polymerase region (L) of the CDV genome was designed. Using this test, out of the 150 analyzed samples, 46 (30.66%) proved to be positive for CDV, indicating that CDV still represents a high risk to the canine population in Hungary. For the phylogenetical analysis, a primer pair that completely encompasses the hemagglutinin (H) gene of the CDV genome was designed. The amplicons of this region were sequenced in both directions using the appropriate primers. Our results indicate that several different CDV genotypes are currently present in Hungary. Nine of the analyzed Hungarian strains turned out to belong to the so-called Arctic group of CDVs, and were most closely related to non-European strains from North America, China and Greenland, as well as to the phocine distemper virus 2 (PDV-2) isolated from Baikal seals (Phoca sibirica). One of the Hungarian strains showed high similarity to other European isolates from Denmark, Germany, Italy and Turkey, as well as to other isolates from geographically more distant regions, such as the USA. Three Hungarian strains seem to join a new cluster that is formed by only a couple of strains, one isolated from a mink in Denmark, and another from a dog in North America. Using a third set of primers, a restriction fragment length polymorphism (RFLP) assay has also been designed for the fast and reliable differentiation of the wild-type CDVs from the vaccine strains.     

Receptive field mechanism in the vertebrate retina

In the catfish retina there are two types of ganglion cells: in one type (type A cell) a spot of light at the center of its receptive field gives rise to a sustained discharge whereas an annulus gives rise to a transient response, and in the other type (type B cell) the response pattern is reversed for a spot and an annulus. Current injected into the horizontal cell induces spike discharges of the ganglion cell very similar to that elicited by a spot of light or by an annulus. In both types of receptive fields, depolarization of the horizontal cell gives rise to a response of the ganglion cell similar to that elicited by a spot of light, whereas hyperpolarization of the cell gives rise to a response of the ganglion cell similar to that elicited by an annulus. Current through a single injecting electrode could drive two types of cells simultaneously. Interaction between a spot of light and an annulus can also be simulated by replacing one light stimulus by current of the proper polarization injected into the horizontal cells. Results suggest that interactions among three neuronal structures, the receptor, the horizontal cell, and the bipolar cell, produce the basic receptive field organization in the channel catfish.     

管道进口空气吸入的临界水深试验研究

制作1∶3比尺的大小水槽模型,试验研究了灌溉管道进口空气吸入的形式及临界水深与管道流速的关系,得出空气吸入的形式及临界水深与水槽内水流的紊动状态有关,且临界水深随流速的增加而增加的结论。相似性分析表明,临界水深在流速较小时大致符合0.2次方准则,而当流速较大时接近流速一致准则。此外,还根据试验结果对日本有关最小水深标准值进行了讨论     

Proliferation, but not growth, blocked by conditional deletion of 40S ribosomal protein S6

Because ribosome biogenesis plays an essential role in cell proliferation, control mechanisms may have evolved to recognize lesions in this critical anabolic process. To test this possibility, we conditionally deleted the gene encoding 40S ribosomal protein S6 in the liver of adult mice. Unexpectedly, livers from fasted animals deficient in S6 grew in response to nutrients even though biogenesis of 40S ribosomes was abolished. However, liver cells failed to proliferate or induce cyclin E expression after partial hepatectomy, despite formation of active cyclin D-CDK4 complexes. These results imply that abrogation of 40S ribosome biogenesis may induce a checkpoint control that prevents cell cycle progression.     

Marker-assisted selection for two dominant powdery mildew resistance genes introgressed into a hybrid grape population

Molecular marker-assisted selection (MAS) was carried out to track the inheritance of Run1 and Ren1, two dominant powdery mildew resistance genes that originate from different genetic resources. The Run1 locus was introgressed from a Muscadinia rotundifolia × Vitis vinifera BC₄ hybrid plant derived from a recurrent pseudo-backcross breeding scheme, whereas the Ren1 locus was introgressed from the powdery mildew-resistant V. vinifera L. variety Kishmish vatkana. Introduction of the Ren1 resistance gene of V. vinifera origin into the breeding program makes possible the long-term defence of the dominant Run1 gene. Using a BC₅ (BC₄ × Kishmish vatkana) hybrid progeny consisting of 441 plants and applying several SSR and BAC-derived CB markers, we demonstrated that the powdery mildew-resistant phenotype co-segregated with the presence of at least one resistance locus-linked marker in the genome. Our data also corroborated earlier findings that the M. rotundifolia-derived Rpv1 and Run1 loci are closely linked. To further streamline the selection process, we developed a multiplex PCR- and agarose gel electrophoresis-based method for the simultaneous detection of both Run1 and Ren1. The results illustrate that MAS offers a rapid and accurate method to select hybrid genotypes that combine multiple loci of interest in grape.     

Cloning and characterization of Photobacterium damselae ssp. piscicida phospholipase: an enzyme that shows haemolytic activity

A phospholipase gene of Photobacterium damselae ssp. piscicida (ppp) was cloned from a genomic library and its nucleotide sequence was determined. The open reading frame consisted of 1218 bp encoding a protein of 405 amino acids with a predicted molecular mass of 46 kDa. The PPP had identities (53-55%) with phospholipase and haemolysin of Vibrio spp., while it showed low identities (23-26%) with glycerophospholipid cholesterol acyltransferase of Aeromonas spp. A recombinant PPP (rPPP) with a His tag at the C-terminus expressed in Escherichia coli and purified showed phospholipase activity. The rPPP also showed lecithin-dependent haemolytic activity against mammalian erythrocytes and direct haemolytic activity against fish erythrocytes. The culture supernatant of wild-type P. damselae ssp. piscicida showed phospholipase activity, while that of a PPP gene knockout mutant did not.     

Diversity of fimbrillin among Porphyromonas gulae clinical isolates from Japanese dogs

Porphyromonas gulae, a gram-negative black-pigmented anaerobe, is a pathogen for periodontitis in dogs. An approximately 41-kDa fimbrial subunit protein (FimA) encoded by fimA is regarded as associated with periodontitis. In the present study, the fimA genes of 17 P. gulae strains were sequenced, and classified into two major types. The generation of phylogenetic trees based on the deduced amino acid sequence of FimA of P. gulae strains along with sequences from several strains of Porphyromonas gingivalis, a major cause of human periodontitis, revealed that the two types of FimA (types A and B) of P. gulae were similar to type I FimA and types II and III FimA of P. gingivalis, respectively. A PCR system for classification was established based on differences in the nucleotide sequences of the fimA genes. Analysis of 115 P. gulae-positive oral swab specimens from dogs revealed that 42.6%, 22.6%, and 26.1% of them contained type A, type B, and both type A and B fimA genes, respectively. Experiments with a mouse abscess model demonstrated that the strains with type B fimA caused significantly greater systemic inflammation than those with type A. These results suggest that the FimA proteins of P. gulae are diverse with two major types and that strains with type B fimA could be more virulent.     

Cutaneous papilloma and multicentric squamous cell carcinoma in four-toed hedgehogs (Atelerix albiventris)

Skin lesions possibly caused by Papillomavirus infections in two four-toed hedgehogs are described. In case 1, there was a papillary mass on the right hind limb. Histologically, the mass was consistent with a viral papilloma. In the other case, multifocal papillary masses with erosions and ulcers were found throughout the body, mainly on the extremities. Histology showed continuative lesions composed of acanthosis, Bowenoid in situ carcinoma, and squamous cell carcinoma, with abrupt transitions between the lesions. In both cases, keratinocytes in the granular layer infrequently had features of koilocytes and intranuclear inclusion bodies, and immunohistochemical staining was positive for anti-human papillomavirus antibody. To the best of the authors’ knowledge, this is the first pathological documentation of possibly papillomavirus-associated skin lesions in four-toed hedgehogs.