Genetic characterization of a novel bovine papillomavirus member of the Deltapapillomavirus genus

This report describes the complete genomic sequence and taxonomic position of BPV type 13. The BPV13 genome was amplified using the multiply primed rolling-circle amplification technique and long-template PCR employing two specific primers. The two long PCR fragments obtained were cloned and sequenced via primer walking. The complete genomic sequence of the BPV13 contains 7961 bp encoding eight proteins, E1, E2, E4, E5, E6, E7, L1, and L2. Similarly to the E5 gene in BPVs 1 and 2, the putative BPV13 E5 ORF encodes a small transforming protein that contains a hydrophobic transmembrane domain. Meanwhile, the retinoblastoma tumor suppressor-binding domain is absent in the putative BPV13 E7 protein. The presence of these two specific molecular features has been recognized as a distinct marker for the development of fibropapilloma in artiodactyl PV-induced lesions. The phylogenetic analysis demonstrated that BPV13 is a new member of the Deltapapillomavirus genus, to be classified as the third representative of the Delta 4 species. The characterization of the genomic sequence of this novel PV will aid in the interpretation of the pathologies described to be related to this virus and provide support for the development of diagnostic tools for epidemiological surveillance of BPV13 in its potential natural hosts.     

Soil type as a putative risk factor of ovine and caprine paratuberculosis seropositivity in Spain

Relationships between soil type and ovine and caprine paratuberculosis in the Avila region (central Spain) were evaluated using data from a cross-sectional study of the most-important diseases of small ruminants in this Spanish region between 1996 and 1997. Questionnaire data from 61 herds (38 ovine and 23 caprine) and 1451 serum samples (1041 ovine and 410 caprine) were used. Herd paratuberculosis (herds were scored as positive to paratuberculosis if any of the serum samples was positive in an agar-gel immunodifussion) was the outcome of interest, whereas soil type in the municipality where farms were located was the predictor variable. Other variables related to soil and soil usage, and herd size, replacement, main food production and animal species were also introduced into the multivariable logistic regression. The final model contained only two independent variables: the predictor variable soil type (coded as two dummy variables ST-1 and ST-2) and herd size (dichotomized at the highest quartile). The estimated Odds Ratios were 25.9 (95% CI: 1.6, 411) for ST-1 (entisols as soil type) and 3.5 (95% CI: 0.3, 45) for ST-2 (inceptisols as soil type).     

Genetic study of Andalusia's ovine and caprine breeds

SUMMARY: Two different breeds of Andalusian sheep, 'Grazalema Merino' and 'Lebrijan Churro', and two different breeds of Andalusian goats, 'Andalusian White' and 'Andalusian Black', chosen by previous studies (Rodero et al. 1992a) as priority breeds for conservation, were studied. The systems used corresponded to ethnozootechnic characteristics, as well as the different biochemical-polymorphism variables. Farms were differentiated within breeds, or between themselves, and different tests were used of genetic and genotypic frequencies: Wright's indices, medium heterozygosities, Whalund's variances, G test of probability of reason, etc. Also Cavalli-Sforza's genetic distance was obtained. In the Andalusian Black and Grazalema Merino breeds, the Whalund's variances obtained were a result of selection, that has divided the breeds into distinct populations differentiated spatially. Medium heterozygosities of each breed do not differ much within themselves, but when each system is considered alone, discrepancies between ethnic groups are relevant. Wright's F indices demonstrated in the Andalusian White and Grazalema Merino breeds, genetic heterozygosities between populations or studied herds can be deduced, but this is not possible in the Andalusian Black. The F(IS) values indicated, despite the small size of the populations, that inbreeding has been avoided, probably because of the entry of foreign sires. In none of the breeds is there a significant excess of heterozygosis. The genetic distances between flocks within breeds do not differ from those found between breeds. RéSUMé: On a travallé avec, differents troupeau des races de montons de l'Andalusie, Grazalema Merino et Lebrija Churro, et avec les races caprines Andalusian White et Andalusian Black, choisie entre les races Andaluciennes comme prioritaires pour la conservation, dans un etudie avant (Rodero et col. 1992a). Les sistémes utilicés dans cette travaille correspondent á charactérés etnozootechniques et á differents variables de polymorphism biochimique. Lorsque on fait differences entre troupeau, dedans de races, ou entre elles, on a utilicés differents preuves, á partir des fréquences géniques et génotipiques: l'index de Wright, hétérozygotie moyennes, variances de Whalund, preuve G de raison de probabilité, etc. Aussi le distance de Cavalli-Sforza. Comme conclusion, dans les races Andalusian Black et Grazalema Merino les variances de Whalund obtenues sont cosequences de l'action de la selection, donant different populations avec differentiation spaciale. Les hétérozygoties moyennes de chaque race sont parus, mais lorsque on considérent chaque systéme separé, les differences entre groupes ethniques sont importantes. Les indexes F de Wright demonstrent que, dans le races Andalusian White et Grazalema Merino on peuvent déduire d'heterozygoties génétique entre les populations ou troupeau analicées, dans le race Andalusian Black les differences valeurs de FIS indiquent que, malgré les petites dimensions des populations, on a evité la consanguinitée, due, probablement, á l'entrée d'étalons externes. Il n'y a pas, chez auqune race, d'un signifivative accroissement d'hétérizygosis. Les distances génétiques entre troupeau, dedans des races, en different pas des distances obtenues entre races. RESUMEN: Se ha trabajado con diferentes ganaderias de las razas ovinas andaluzas Merino de Grazalema y Churra Lebrijana, y con las caprinas Blanca Serrana y Negra Serrana, elegidas entre el resto de las razas de Andalucia como prioritarias pra la conservación, por estudio previo (Rodero y col., 1992a). Los sistemas utilizados en este trabajo corresponden tanto a caracteres etnozootécnicos como a diferentes variables de polimorfismos bioquimicos. Cuando se han diferenciado las ganaderias dentrde razas, o las ganaderias entre si, se han utilizado diferentes pruebas, a partir de las frecuencias genéticas y genotipicas: indices de Wright, heterocigosidades media, varianzas de Whalund, prueba G de razón de probabilidad, etc. También se obtuvieron las distancia genéticas de Cavalli-Sforza. Se concluye que en las razas Negra Serrana y Merino de Grazalema las varianzas de Whalund obtenidas son consecuencia de la acción de la selección que ha actuado dividiendo las razas en distintas poblaciones con diferenciación espacial. Las heterocigosidades medias de cada raza no difieren mucho entre si, pero cuando se considera cada sistema aisladamente, las discrepancias entre grupos étnicos son acusadas. Los indices F de Wright ponen de manifiesto que, mientras en las razas Blanca Serrana y Merino de Grazalema se pueden deducir heterocigosidades genéticas entre las problaciones o ganaderias estudiadas, no ocurre otro tanto en la raza Negra Serrana. Los valores de F(IS) parecen indicar que, a pesar del tama?o peque?o de las poblaciones, se ha evitado la consanguinidad, probablemente por la entrada de sementales externos. No se produce en ninguna de las razas un exceso significativo de heterocigosis. Las distancias genéticas entre ganaderias dentro de razas no difieren de lashalladas entre razas. ZUSAMMENFASSUNG: Es wurde mit verschiedenen andalusischen Zuchten der Schafrassen 'Grazalema Merino' and 'Lebrijan Churro' und der Ziegenrassen 'Andalusian White' und 'Andalusian Black' gearbeitet, die man von den andalusichen Rassen im Hinblick auf Erhaltung ausgew?hlt hat. Die benutzten Systeme in dieser Forschungsarbeit entsprechen Merkmalen, die sich sowohl auf ethnisch wie auch auf die verschiedenen Variablen des biochemischen Polimorfismius bezichen. Zur Unterschlidung von Zuchten innerhalb die Rassen oder von zuchten untereinander wurden verschiedene Tests benutzt, die von den genetischen und genotypischen Frequenzen ausgehen: Wright-Index, Durchschnittsheterozygosit?ten, Wahl und Varianz G-Test der Wahrscheinlichkeit, etc. Au?erdem wurden die genetischen Distanzen nach Cavalli-Sforza errechnet. Man Kommt zum Schlu?, da? in den Andalusian Black und Grazalema Merino die Wahl und Varianz das Ergebnis einer Selektionsaktivit?t ist, die die Rassen in Verschiedenen Populationen und unterschiedlichen R?umen aufgeteilt hat. Die durchschnittlichen Heterozygosit?ten jeder Rasse unterscheiden sich wenig voneinander, aber wenn man jedes System für sich betrachtet, stell man doch erhebliche Diskrepanzen zwischen den ethnischen Gruppen fest. Der Wright-Index offenbart, da? man in den Rassen Andalusian White und Grazalema Merino genetische Heterozygosit?ten ableiten kann zwischen den Populationen oder den untersuchten Zuchten; dies ist nicht der Fall inder Rasse Andalusian Black. Die F(IS) werte scheinen anzugeben, da trotz der kleinen Gr??e der Populationen die Blutsverwandschaft vermieden wurde, wahrscheinlich durch von au?erhalb kommenden B?cken. In keiner der Rassen existiert überm?ssige Heterozygotie Die genetischen Distanzen zwischer der Zuchten unterscheiden sich nicht van dener der Rassen.     

J亚群与E亚群禽白血病自然重组病毒的全基因组序列分析

为了解我国东北地区部分养鸡场禽白血病病毒(ALV)的基因组序列特征及其变异情况,本研究从具有典型血管瘤病变的禽白血病发病鸡中分离到一株J亚群ALV(ALV-J)命名为JL0901,并进行了全基因测序.将该序列与已发表的ALV-J毒株序列进行比较研究,结果表明JL0901基因组的gag和pol基因相对保守,而env基因和3'端非编码区(3'UTR)的变异较大.对JL0901的env基因核苷酸序列进一步分析发现,在其gp85基因和gp37基因交界位置发生J亚群和E亚群ALV重组现象.本研究证实国内鸡群中存在J亚群和其他亚群ALV的自然重组现象,并表明国内ALV已出现新的变异趋势.     

Molecular and biological characterization of a naturally occurring recombinant subgroup B avian leukosis virus with a subgroup J-like long terminal repeat

Infection of broiler chickens with subgroup J avian leukosis virus (ALV) results in the induction of myeloid tumors. However, although egg-type chickens are susceptible to infection with ALV-J, the tumor incidence is very low, and on rare occasions the tumors observed are of the myeloid lineage. We recently described the isolation of an ALV (AF115-4) from commercial egg-type chickens suffering from myeloid leukosis. AF115-4 was initially identified as an ALV-J isolate based on PCR analysis of the long terminal repeat (LTR). However, further characterization of the viral envelope indicated that the virus is recombinant with subgroups B envelope and J LTR. Here we further characterize this recombinant virus at both the molecular and biological levels. We show that the AF115-4 isolate expresses a recombinant envelope glycoprotein encoded by a subgroup B gp85 region and a subgroup E gp37 region. The host range ofAF115-4 was analyzed using cells resistant to infection by subgroups A/B, J, or E; this shows that no ALV-J was present in the isolates obtained from the affected chickens. Additional antigenic characterization of AF115-4 using chicken sera specific for subgroups B or J indicated that no ALV-J was present in the samples examined. Inoculation of AF 115-4 into ALV-susceptible 1515 X 71 chickens resulted in the induction of lymphoid leukosis but not the expected myeloid leukosis affecting the commercial chickens. These results suggest that differences in the genetic makeup of the chickens from which AF115-4 was isolated and the line 1515 X 71 used in the present experiments may be responsible for the observed differences in pathogenicity. In addition, the results suggest that ALV-J continues to evolve by recombination, generating new viruses with different pathological properties.     

The first identification of contagious caprine pleuropneumonia (CCPP) in sheep and goats in Egypt: molecular and pathological characterization

Tropical Animal Health and Production - Contagious caprine pleuropneumonia (CCPP) is one of the most fatal and contagious diseases of goats. To date, the occurrence of CCPP in Egypt has not been...     

Dietary molybdenum as a putative copper antagonist in the horse

Four horses were stabled and fed a diet of hay ad libitum, and 2 kg oats per animal per day, for a month. The basic diet was then supplemented with molybdenum, at a rate of 20 mg/kg dry matter for 4.5 months. For one month of this period the diet was supplemented also with sulphur at a rate of 1.2 g/kg dry matter. Analyses of jugular blood samples, obtained at intervals varying between two and 20 days, showed no evidence of a decline in total plasma copper or of an increased proportion of trichloroacetic acid (TCA) insoluble copper in plasma over this period. In separate studies, two other horses were given 99molybdenum (molybdate, 20 to 28 mg Mo, 4 mCi per animal) per os, initially while being fed the basic diet and later while maintained on the molybdenum supplemented diet. 99Molybdenum appeared rapidly in plasma, but the radioactivity was then quickly cleared (half-life 7 to 10 h). The 99molybdenum present was identified as (99Mo)-molybdate. There was no evidence of the persistent, protein-bound (99Mo)thiomolybdates which appear in ruminants. These studies indicate that increased dietary molybdenum is unlikely to interfere with copper metabolism in horses.     

Genetic characterization of a novel influenza A virus H5N2 isolated from a dog in China

An influenza virus, A/canine/Shandong/JT01/2009, has been isolated from a dog exhibiting classical flu signs in China. HAI and NAI assays subtyped A/canine/Shandong/JT01/2009 as a H5N2 like virus. Phylogenetic reconstructions indicated strong relationships with viruses from various hosts and dispersed geographic locations. These analyses indicate A/canine/Shandong/JT01/2009 is a novel virus generated by complex reassortment of the viral segments.     

1株HoBi样瘟病毒的分离与序列分析

本研究在MDBK细胞培养物中发现并分离出1株HoBi样瘟病毒,将该毒株命名为JS12/01。应用RT-PCR方法分段扩增该毒株的全基因组序列,并对其基因组序列进行分析。结果显示,该毒株基因序列2端分别为5′端非编码区(5′untranslated region,5′-UTR)和3′端非编码区(3′untranslated region,3′-UTR),编码区为11 700nt。与GenBank中登录的部分瘟病毒属代表毒株5′-UTR区序列进行进化分析,结果表明,该毒株属于HoBi样瘟病毒巴西源亚型。本研究中的HoBi样瘟病毒JS12/01株为国内首次分离鉴定,证实了该病毒在中国的存在,需要对该病毒的扩散与流行采取预防措施。     

饲料添加剂中的新成员——益生酸的应用研究（上）

饲料添加剂一般可促使动物增重快，产奶、产蛋率高，发展牧业生产，除饲料，管理外，更主要的是防疫工作，一旦发生疾病，损失惨重，饲料添加剂的新成员——益生酸，是农副产品下脚料加以辅料，通过低温，中温和高温多类，多种菌群自然发酵和萃取而成的生物高新科技产品，具有添加量小，效果好等特性，除增重，产奶量高，产蛋多外，还可使畜（畜）免疫力提高，发病少，对一些疾病有治疗作用，抗逆性好，成活率高，其肉类，奶类及蛋类产品可达到无公害标准，提高经济效益。     

Pathogenicity of a caprine herpesvirus.

A herpesvirus isolated from neonatal Angora kids (Capra hircus) with a relatively severe generalized infection was shown to be infective for adult goats as well as for kids. However, the virus lacked pathogenicity for either lambs or calves. Although a nonreciprocating serologic overlap exists between the caprine virus and the antibody to bovine herpesvirus (BHV-1), results of cross immunity tests in calves indicated that the 2 viruses are immunologically distinct. On the basis of these findings, the caprine virus seems to be a specific pathogen of goats. Accordingly, the designation of Herpesvirus caprae or caprine herpesvirus 1 seems taxonomically appropriate.     

Characterization of a caprine herpesvirus.

A virus, which was isolated from kids (Capra hircus) affected with a relatively severe generalized infection, was found to contain DNA and to have a buoyant density of 1.2820 g/cm3. The virus was sensitive to the action of lipid solvents and trypsin and was rapidly inactivated at pH 3.0 and at temperatures of 50 and 56 C. The virion, an icosahedron consisting of a nucleoid surrounded by a double membrane, measured approximately 135 nm in diameter. On the basis of its chemical and physical properties, the virus is considered a herpesvirus.     

Genetic and splice variations of Bos taurus CD46 shift cell permissivity to BVDV, the bovine pestivirus

The pestivirus bovine viral diarrhea virus (BVDV) is known to bind to the CD46 molecule, which subsequently promotes entry of the virus. Mapping of the BVD-virion-binding site has shown that two peptides, 66EQIV69 and 82GQVLAL87, located on antiparallel beta sheets in the most distal complement control protein module (CCP1), provide the attachment platform. In the present study, we reveal the existence of ten distinct allelic versions of the CCP1 module, varying significantly in frequency among taurine and indicine races. A complex mRNA splicing pattern was also evidenced for bovine CD46, generating three different serine-threonine-proline segments and five different cytoplasmic domains. The four most frequent allelic variants and the six splice variants were then expressed in BVDV-nonpermissive porcine cells and the quantity of progeny virions generated by each cell preparation was measured 48 h post-infection. As expected, ectopic expression of the 10 bovine CD46 isoforms rendered the PK15 cells permissive to BVDV, as attested by the 100,000-fold greater recovery of virions from these cells than from non-transfected cells. This permissivity increase was significantly lower (-33%, P<0.001) when the canonical CCP1 was replaced with the variant most frequent in zebus, suggesting positive or negative selection of this allele in the latter and in the former, respectively. The predicted secondary structure of this variant suggests that the measured loss of function is due to the disappearance of one of the two beta sheets constituting the BVDV attachment platform. On the other hand we showed that for a given CCP1, the titer recovered at 48 hpi also depended on the nature of the CD46 cytoplasmic domain (P<0.001). This result implies that virus binding generates a cytoplasmic-tail-dependent outside-in signal that determines permissivity to BVDV.     

Isolation of Chuzan virus, a new member of the Palyam subgroup of the genus Orbivirus, from cattle and Culicoides oxystoma in Japan

Five virus strains with identical antigenic properties were isolated from 3 RBC suspensions obtained from 2 healthy sentinel calves and from 2 pools of Culicoides oxystoma in cultures of a hamster lung cell line (HmLu-1). The virus was tentatively named Chuzan virus. The Chuzan virus was classified as a new member of the Palyam subgroup of the genus Orbivirus on the basis of its physicochemical, morphologic, and antigenic properties.     

Transmission of a pestivirus infection in a population of Pyrenean chamois

Outbreaks of a previously unrecorded disease have recently affected Pyrenean chamois (Rupicapra pyrenaica pyrenaica) populations across the mountain range. A pestivirus was hypothesized to be the cause of this emerging disease and this type of virus can cross the species barrier and be transmitted to or from wildlife. Using an epidemiological survey conducted from 1995 to 2004 at Orlu, France, we characterized the virus and analyzed its transmission. A phylogenetic analysis of viral sequences and virus neutralization tests showed that the virus belonged to the newly described border disease virus-4 group. The increase of seroprevalence with age indicated that infection can occur at any age and resulted in lifelong immunity. Overall, 70.3% of 323 samples were positive for anti-p80 antibodies and 10.2% of 167 samples showed viremia, as demonstrated by either positive ELISA antigen test or RT-PCR. Infection has thus been widespread in this population since 1995, whereas no mass mortality or clinical signs have been observed. Incidence and seroprevalence varied seasonally and according to number of individuals aged less than 2 years old in the population, so viral transmission was dependent on host population age structure. We propose that the virus is now endemic in this population and is likely detrimental for reproduction and juveniles. Further investigation is needed to estimate the impact of pestivirus on host population dynamics and the risk of cross-transmission to farm animals.     

蛋鸡J亚群禽白血病病毒SD1009分离株的分离鉴定及全基因组序列分析

为分析我国J亚群禽白血病病毒(ALV-J)蛋鸡分离株的进化关系,本研究将山东省某鸡场采集的蛋鸡病料样品接种DF-1细胞系,利用ELISA群特异性抗原检测以及亚群特异性间接免疫荧光方法,分离鉴定得到一株ALV-J,命名为SD1009,并对其进行全基因组测序,将该序列与其他ALV-J代表性病毒株序列进行比较。结果表明:SD1009分离株的gag和pol基因相对保守,与各参考病毒株的同源性为95%～99%,env基因的同源性为91%～95%;在其5'UTR中出现了连续19 bp的插入突变,与TW-3577、SDAU09C3、JS09GY6、JS09GY3蛋鸡分离株的5'UTR基本一致,提示19 bp的插入现象可能是近年来蛋鸡ALV-J的进化趋势;此外,其3'UTR的rTM和DR区出现部分缺失现象,该缺失部分也可能与ALV-J进化相关。