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1.
Objective
Characterisation of a complete genome sequence of an Australian strain of canid alphaherpesvirus 1 (CHV‐1) and its phylogenetic relationship with other varicellovirus species.Methods
Standard pathology and PCR methods were used to initially detect herpesvirus in hepatic tissue from an infected 4‐week‐old Labrador Retriever puppy. The complete CHV‐1 genome was sequenced using next‐generation sequencing technology followed by de novo and reference assembly, and genome annotation.Results
The CHV‐1 genome was 125 kbp in length and contained 74 predicted open reading frames encoding functional proteins, all of which have counterparts in other alphaherpesviruses. Phylogenetic analysis using the DNA polymerase gene revealed that the newly sequenced CHV‐1 clustered with canid alphaherpesvirus isolated from the UK and shared a 99% overall nucleotide sequence similarity.Conclusion
This is the first complete genome of an Australian strain of CHV‐1, which will contribute to our understanding of the genetics and evolution of herpesvirus. 相似文献2.
X N Xuan T Horimoto M Ono J A Limcumpao Y Tohya M Azetaka E Takahashi T Mikami 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(6):1181-1188
Eighteen canine herpesvirus (CHV) isolates from Japan and two reference strains were compared by restriction endonuclease analysis technique using total DNA extracts from cells infected with the viruses. In order to select the suitable restriction endonucleases for differentiation of CHV isolates, ten enzymes were used and three of them, HindIII, XbaI, and PvuII, were found to be useful for strain differentiation. With these enzymes, CHV isolates from unrelated individuals were readily differentiated from each other. In contrast, all the isolates derived from the same litter were not distinguishable on the basis of restriction cleavage patterns. However, slight mobility shifts were observed among the isolates from the same litter or the same individual. The results showed that this method provides a powerful tool for epidemiological surveys of CHV infection. 相似文献
3.
The effects of canine herpesvirus (CHV) on fetuses were studied after IV inoculation of pregnant bitches in the 2nd trimester of gestation. Cesarean sections were performed on 2 bitches that were inoculated with CHV on the estimated 30th day of gestation. Bitch M-1 had 2 mummified fetuses and bitch M-2 had 4 mummified and 2 dead fetuses and 3 live-born pups. Infection by CHV was confirmed histopathologically by the presence of focal areas of necrosis associated with intranuclear inclusion bodies in heart muscle sections of 1 dead fetus; CHV was not recovered from other organs. Abortion occurred between the 2nd and 3rd week after inoculation of another pregnant bitch inoculated with CHV on the estimated 30th day of gestation. Two bitches inoculated with CHV on the estimated 40th day of gestation gave birth prematurely to 10 pups. The detection of characteristic herpesviral lesions in various organs and the reisolation of CHV from the liver, spleen, kidneys, and lungs of premature pups indicated CHV infection. Transplacental infection of fetal pups by CHV resulted in their death and subsequent mummification. It appears that abortion and premature birth also may occur in pregnant bitches infected during the 2nd trimester of gestation. 相似文献
4.
A Hashimoto K Hirai K Okada Y Fujimoto 《American journal of veterinary research》1979,40(9):1236-1240
Pathologic and virologic investigations were done on the fetal placenta and on pup runts which were obtained from a bitch with a medical history of canine herpesvirus (CHV) infection. Macroscopically, the placenta was poorly developed. Small grayish white foci were observed in the placental labyrinth. Characteristic lesions of CHV infection were not prominent in the pups examined. Microscopically, however, focal degenerative and necrotizing lesions were observed in the placental labyrinth. Rarely, eosinophilic or basophilic intranuclear inclusion bodies were in the trophoblastic cells in the necrotizing lesions. In the adrenal gland of one stillborn pup, focal necrosis and hemorrhages could be seen; these irregularities were essentially the same as those seen in the newborn pups with CHV infection. Focal interstitial pneumonia was also observed in some of the pups. The CHV organism was isolated from the kidney of one pup that survived for 22 days. 相似文献
5.
Canine Herpesvirus (CHV) is being developed as a virus vector for the vaccination of European red foxes. However, initial studies using recombinant CHV vaccines in foxes revealed viral attenuation and lack of antibody response to inserted foreign antigens. These findings were attributed both to inactivation of the thymidine kinase (TK) gene and excess foreign genetic material in the recombinant viral genome. In this study, we report an improved CHV-bacterial artificial chromosome (BAC) vector system designed to overcome attenuation in foxes. A non-essential region was identified in the CHV genome as an alternative insertion site for foreign genes. Replacement of a guanine/cytosine (GC)-rich intergenic region between UL21 and UL22 of CHV with a marker gene did not change growth behaviour in vitro, showing that this region is not essential for virus growth in cell culture. We subsequently produced a CHV-BAC vector with an intact TK gene in which the bacterial genes and the antigen expression cassette were inserted into this GC-rich locus. Unlike earlier constructs, the new CHV-BAC allowed self-excision of the bacterial genes via homologous recombination after transfection of BACs into cell culture. The BAC-CHV system was used to produce a recombinant virus that constitutively expressed porcine zona pellucida subunit C protein between the UL21 and UL22 genes of CHV. Complete self-excision of the bacterial genes from CHV was achieved within one round of replication whilst retaining antigen gene expression. 相似文献
6.
A Takumi K Kusanagi K Tuchiya X N Xuan M Azetaka E Takahashi 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(2):241-250
An enzyme-linked immunosorbent assay (ELISA) for the diagnosis of canine herpesvirus (CHV) infection using antigen prepared by solubilizing infected cells was developed. The ELISA and two improved methods of serum neutralization test, the microplate serum neutralization test (MSNT) with complement and the 50% plaque reduction (PR) assay with complement, were compared for the results of antibody detection from a total of 557 field canine sera. Of 529 sample sera that were negative in the MSNT with complement, 119 were ELISA positive, and this result together with time course of serum antibody detection in a dog experimentally infected with CHV strongly suggested that the MSNT with complement is less sensitive for the detection of antibody in CHV infected dogs, especially those in early stages of infection. A correlation was found between the titers measured by the ELISA and 50% PR assay with complement, however, for field use, the ELISA is recommended as a highly sensitive test method of serodiagnosis of CHV infection adequate for dealing with a large number of samples with less demand on time and effort. 相似文献
7.
The alphaherpesvirus canine herpesvirus (CHV) was tested in order to determine whether or not it has apoptotic potential. We have demonstrated that lytic replication of CHV resulted in induction of apoptosis. This phenomenon was confirmed using different techniques including in situ TUNEL assay and DNA laddering. The apoptotic activity of CHV might influence the pathobiology of this virus. 相似文献
8.
Studies were conducted to evaluate the feasibility of using canine herpesvirus (CHV) as a vaccine vector for bait-delivered oral vaccination of wild foxes. To test the viability of CHV in baits, CHV was freeze-dried, incorporated into different baits, stored, and the remaining viral infectivity tested in cell culture after varying periods of time at different storage temperatures. Experimental baits (mouse carcasses) and commercial baits (FOXOFF and PROBAIT) were prepared with either liquid or freeze-dried CHV and tested in two fox trials for their capacity to induce CHV-specific antibodies following oral baiting. Freeze-drying and storage temperatures below 0 degrees C had a stabilizing effect to virus infectivity. When stored at -20 degrees C, freeze-dried CHV retained its full infectivity for up to 3 months in PROBAIT baits, the remaining infectivity in FOXOFF baits was 100-fold less. Oral baiting with CHV induced antiviral serum antibodies in all vaccinated foxes (20/20). None of the vaccinated foxes became ill or shed infectious virus into the environment although viral DNA was detected in body secretions as evaluated by PCR. The results indicate that CHV can be freeze-dried and stored over extended periods of time without loosing much of its infectivity. This is the first report of CHV being used for oral bait vaccination of foxes. It appears that CHV is well suited for use as a recombinant vector for wild canids. 相似文献
9.
Reubel GH Pekin J Venables D Wright J Zabar S Leslie K Rothwell TL Hinds LA Braid A 《Veterinary microbiology》2001,83(3):217-233
We report on the pathogenicity of canine herpesvirus (CHV) for European red foxes. In the first experiment, we inoculated 10 adult foxes intravenously with a canine isolate of CHV. All foxes became infected and shed CHV in saliva and genital secretions for up to 14 days post-inoculation (p.i.) as evaluated by PCR and/or by virus isolation. All foxes developed clinical signs such as fever, lethargy and evidence of respiratory tract disease. Two foxes died on day 6 p.i., one on day 7 p.i., and one fox was euthanased on day 6 p.i. Tissues taken from the four dead foxes were positive for CHV by PCR. The remaining six foxes recovered after approximately 14 days p.i. Virus particles with morphology typical of herpesviruses were found by electron microscopy in the liver of an infected animal. All surviving foxes developed serum anti-CHV antibodies. In a second experiment, six foxes were dosed perorally with CHV and paired with six untreated controls. Neither the perorally dosed nor the in-contact control foxes developed clinical signs of disease. Infectious CHV was not isolated from any of the dosed or the in-contact foxes but all perorally-infected foxes and one of the in-contact foxes tested PCR-positive for CHV on several occasions p.i. All perorally-infected foxes, but none of the in-contact foxes, seroconverted. In summary, intravenous CHV inoculation caused a clinical disease in adult foxes much more severe than observed in experimentally-infected adult dogs. No clinical disease or virus spread was observed after peroral dosing although viral infection occurred as evidenced by seroconversion. 相似文献
10.
Anette Krogen?s Vibeke Rootwelt Stig Larsen Lena Renstr?m Wenche Farstad Arve Lund 《Acta veterinaria Scandinavica》2014,56(1):19
Background
Canine herpesvirus-1 (CHV1) causes a fatal hemorrhagic disease in neonatal puppies and is associated with infertility in female dogs. This study was conducted to assess the status of CHV1 infection in bitches in proestrus or estrus and to investigate possible risk factors by a detailed questionnaire. Blood samples were collected from healthy bitches (n = 193) not vaccinated against CHV1, aged one year or older and admitted for estrus control to the Canine Reproductive Clinical Unit, Norwegian School of Veterinary Science. The serum samples were analysed by immunoperoxidase monolayer assay and serum titers were recorded as the reciprocal value of the highest dilution producing specific cell staining.Results
Altogether, 85.5% of the dogs had CHV1 titers ≥ 80 and were classified as positive. Mean age for dogs included in the study was 4.2 years (95% CI 4.0-4.5), and there was no difference in age between seronegative dogs vs seropositive dogs. When grouping the seropositive dogs into three categories according to the magnitude of the titer, a total of 38.8% of the bitches displayed a weakly positive titer of 80, 44.8% had moderately positive titers of 160 or 320 and 16.4% of the dogs fell into the strongly positive category with titer of ≥640. No association was demonstrated when comparing CHV1 antibody titers to fertility parameters such as previous matings, pregnancies, whelpings, puppies born or condition of puppies. Further, there was no difference in seroprevalence between bitches that had been abroad for a period of time and dogs only living within a Norwegian environment. Samples from dogs collected in summer and fall displayed moderate to high antibody titers indicating recent infection with CHV1. Season, previous birth, and participation in competitions/shows explained 67-78% of the variation in antibody titer.Conclusions
This study demonstrates that CHV1 infection is common in breeding bitches in the eastern part of Norway. Associations with putative risk factors were not identified. However, season, previous whelping, and participation in competitions/shows explained 67-78% of the variation in antibody titer. 相似文献11.
12.
Yoon HA Eo SK Aleyas AG Cha SY Lee JH Chae JS Jang HK Cho EG Song HJ 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(2):143-148
The prevalence and quantity of latent pseudorabies virus (PrV) in nervous tissues of pigs exposed to field strain in Korea was investigated by nested and real-time PCR. Nervous tissues including trigeminal ganglion (TG), olfactory bulb (OB), and brain stem (BS) were collected from 94 seropositive pigs. PrV latent infection in nervous tissues was initially investigated by nested PCR targeting three glycoprotein genes (gB, gE, and gG). Based on the obtained result, latent infection was detected in 95.7% of screened animals. Furthermore, it was revealed that the examined tissues harbored different copy numbers of latent PrV genome ranging from <10(2.0) to 10(7.1) copies per microgram of genomic DNA in real-time PCR analysis. These results show that under normal conditions, levels of latent PrV in the nervous tissues of pigs can vary across a wide range. Therefore, the data presented here provides information regarding control of the endemic state of PrV in Korea. 相似文献
13.
Detection of latent pseudorabies virus in porcine tissue, using a DNA hybridization dot-blot assay 总被引：2，自引：0，他引：2
R G McFarlane D G Thawley R F Solorzano 《American journal of veterinary research》1986,47(11):2329-2336
A DNA-hybridization dot-blot technique was developed to detect the presence of pseudorabies virus (PRV) DNA in porcine tissue. Seven 32P-nick translated probes of high specific activity were prepared from transformed Escherichia coli plasmids into which Bacillus amyloliquefaciens H (Bam HI) restriction fragments of PRV-DNA had been inserted. Samples of DNA that had been extracted from porcine tissue or from PRV grown in tissue culture were transferred to nitrocellulose paper, using a microsample filtration manifold and were hybridized to the probes under high-stringency conditions. Under optimal hybridization conditions, the minimum detection amount of PRV-DNA was 10(-11) g, which is equivalent to 1 copy of the PRV genome/80 host cells. Four probes did not show cross hybridization with DNA extracted from tissues of known PRV-negative swine, and these were subsequently used to detect PRV-DNA in infected porcine tissues. Generally, correlation between virus isolation and hybridization data was good for tissues from swine that had died of acute PRV infection. Furthermore, PRV-DNA was present in specific tissues of all 4 seropositive swine that had recovered from pseudorabies and in which no infective virus or viral products were detected at necropsy. Pseudorabies virus DNA was present in the rostralis cerebral cortex (n = 2) or in the medulla oblongata (n = 1) and trigeminal ganglion (n = 1). This probably indicated the portal of entry of the virus into the CNS. In another seropositive pig, there was evidence of a productive infection in the tonsils, although virus was not isolated in a tissue culture system.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
14.
15.
《Veterinary microbiology》1998,62(3):171-183
The presence of Aujeszky's disease virus (ADV) in peripheral blood mononuclear cells (PBMC) and tissues of experimentally infected pigs was studied. Vaccinated and unvaccinated pigs were inoculated with different doses of Aujeszky's disease NIA-3 strain. Pigs were periodically bled and PBMC were used for virus isolation and PCR detection of virus. Tissues were obtained at the time of death (8 weeks post-inoculation) and used for ADV genome detection by PCR. ADV genome was amplified from PBMC during the acute phase of infection and, in some experimental groups, up to 38 days post-inoculation (PI). The virus was sporadically detected by virus isolation performed from PBMC. In neural tissues, ADV was constantly amplified from the trigeminal ganglia and the olfactory bulb of persistently infected pigs (euthanised 8 weeks PI). In other tissues, the viral genome was rarely detected in lymph nodes and tonsils, and, occasionally, in the bone marrow. Our results indicated that PBMC are not an appropriate source for detecting ADV persistence, since inconsistent results were obtained throughout the experiments. In neural tissues, the olfactory bulb turned out to be as important a target for ADV persistence as the trigeminal ganglia. Viral genome detection in the bone marrow indicated that this tissue may play a role in the establishment of a persistent infection. 相似文献
16.
Mycoplasma hyopneumoniae DNA was detected in 20 naturally infected pigs by in situ hybridization using a nonradioactive digoxigenin-labeled DNA probe. A 520-base-pair DNA probe targeting a reiterative sequence of the M. hyopneumoniae genome was generated by the polymerase chain reaction. All 20 pigs infected with M. hyopneumoniae had distinct and positive hybridization signals without background staining. A strong hybridization signal was detected mainly in the luminal surface of bronchial and bronchiolar lining epithelial cells, whereas no hybridization signal was seen in the cytoplasm of bronchial and bronchiolar lining epithelial cells. When hybridization signal was detected in the luminal surface of bronchial and bronchiolar lining epithelial cells, a given bronchus or bronchiole had peribronchiolar lymphoid hyperplastic tissues. Hybridization signals were not seen in the peribronchiolar lymphoid hyperplastic tissues. A less intense signal was detected in the interstitial and alveolar macrophages randomly scattered in the thickened alveolar septa and spaces. Hybridization signal was rarely detected in the type I pneumocytes. The in situ hybridization technique developed in this study was useful for detection of M. hyopneumoniae nucleic acids in tissues taken from naturally infected piglets and may be a valuable technique for studying the pathogenesis of M. hyopneumoniae infection. 相似文献
17.
为了获得狼山鸡性腺轴组织基因组DNA甲基化水平和模式等表观遗传信息,试验采用全基因组重亚硫酸盐测序（whole genome bisulfite sequencing,WGBS）技术检测狼山鸡下丘脑和卵巢组织基因组DNA甲基化状态,分析两组织DNA甲基化水平及特异甲基化模式。结果表明,狼山鸡下丘脑和卵巢基因组整体甲基化水平分别为4.35%和3.48%,差异显著（P<0.05）;下丘脑和卵巢中分别检测到6 150 000和10 320 000个甲基化胞嘧啶（mC）位点,其中mCG类型位点分别占69.99%和87.88%,下丘脑中非mCG位点占比约为卵巢中的2.5倍;与各染色体不同,两组织线粒体基因组中mCHH位点占比最高,其次是mCHG位点;卵巢基因组启动子区DNA甲基化水平极显著低于内含子和外显子区（P<0.01）,极显著高于基因间区（P<0.01）;下丘脑基因组启动子区DNA甲基化水平与内含子和外显子区相比差异不显著（P>0.05）,却显著高于基因间区（P<0.05）;下丘脑基因组各功能元件DNA甲基化水平均显著或极显著高于卵巢基因组（P<0.05;P<0.01）。综上,狼山鸡下丘脑和卵巢组织具有不同的DNA甲基化模式和特征,下丘脑中较高的非mCG位点比例可能在中枢神经系统发育中发挥重要作用,本研究结果为进一步分析鸡卵巢和下丘脑基因组DNA甲基化对其繁殖性能调控机制提供参考依据。 相似文献
18.
Rijsewijk FA Luiten EJ Daus FJ van der Heijden RW van Oirschot JT 《Veterinary microbiology》1999,65(1):1-7
Canine herpesvirus (CHV1) is found in dogs all over the world and may spread by oronasal or sexual contact. We developed an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against CHV1 in dogs. The antigen used for this ELISA was prepared by purifying CHV1 virions from the medium of infected A72 cells. To investigate the prevalence of CHV1 in The Netherlands, a panel of 145 sera of dogs boarding at a kennel in Lelystad, The Netherlands, was screened using this ELISA. The dogs originated from all parts of The Netherlands and represented many different breeds. The sera were collected both at the start and at the end of the boarding period. Of the 145 paired sera 61 (42.1%) were positive, 79 (54.5%) were negative and 5 (3.4%) could not be attributed to either group. None of the negative dogs became seropositive during the boarding period, which lasted normally two to three weeks. We also tested 79 individual sera taken from dogs at various other places in The Netherlands and found that 27 (34.2%) were positive. Hence, in total 224 dog sera, collected from April 1997 to March 1998, were tested and 88 (39.3%) were found positive. We conclude that the prevalence of CHV1 seropositive dogs in The Netherlands in this period was about 40%, and that boarding at a dogs kennel did not contribute to the spread of CHV1. In addition, CHV1 has been isolated from two clinical cases of fatal haemorrhagic disease in The Netherlands. 相似文献
19.
X N Xuan T Horimoto J A Limcumpao K Nemoto Y Tohya M Azetaka E Takahashi T Mikami 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(5):899-905
Monoclonal antibodies (MoAbs) were used to identify the hemagglutinin of canine herpesvirus (CHV). The inhibition of viral hemagglutination (HA) activity was observed with MoAbs against 41 kD glycoprotein, while no hemagglutination-inhibition (HI) activity was observed with those against 145/112 kD and 80 kD glycoproteins, suggesting that the 41 kD glycoprotein is the hemagglutinin of plaque-selected virus of CHV YP11 strain used as immunogen for MoAb production. All of the HI MoAbs also showed HI activities against HA antigens which were prepared from cells infected with other CHV strains, namely, F-205 V and Glasgow CHV2 reference strains, eight Japanese isolates, and the original YP11 strain. However, on immunoblotting analysis, a 47 kD protein band was detected in these strains by the HI MoAbs. These data suggest that the 47 kD glycoprotein is the common molecule of the hemagglutinin among CHV strains and the plaque-selected virus of YP11 strain appears to be a mutant whose molecular weight of the hemagglutinin changed into 41 kD. 相似文献
20.
Viruses with properties consistent with herpesvirus were isolated from dogs with diarrhea. The viruses were shown to be antigenically related to feline herpesvirus-1 (FHV-1) by virus neutralization tests. It was also observed that a canine herpesvirus (CHV) prototype, D004, and two field isolates from fatal CHV infections in 2-week-old and 6-week-old puppies were neutralized at a low level by antiserum to FHV-1. Reciprocal neutralization tests with CHV antiserum against FHV-1 were negative. These results indicated that viruses related to FHV-1 can infect the dog and that there appears to be uni-directional virus neutralization of CHV by FHV-1 antibody. 相似文献