对虾皮下及造血组织坏死杆状病毒的精细结构、核酸、多肽及血清学研究

对纯化的对虾皮下及造血组织坏死杆状病毒（ＨＨＮＢＶ）进行了超微结构、紫外吸收、核酸类型及多肽ＳＤＳ－ＰＡＧＥ的研究。同时用单克隆抗体对ＨＨＮＢＶ的两分离株和ＭＢＶ进行了血清学比较研究。用ＴＭＶ作为内标定物测得病毒粒子大小为１５０ｎｍ×４５０ｎｍ，其衣壳为两个帽状物端夹１３圈螺旋对称的园柱体结构。病毒在２６０ｎｍ处吸光系数为０．１１７ｍｇ／ｍＬ／ＯＤ２６０。病毒核酸为双链ＤＮＡ，分子量大于３５×１０６ｄ。病毒囊膜有１２种主要多肽。病毒衣壳有两种主要多肽。从寿光（ＨＨＮＢＶ—９３７）和青岛（ＨＨＮＢＶ—９３８）分离到的病毒种在ＳＤＳ—ＰＡＧＥ和抗ＨＨＮＢＶ—９３７单抗ＥＬＩＳＡ反应上没有显著差异。ＨＨＮＢＶ与斑节对虾杆状病毒（ＭＢＶ）在抗原决定簇上存在差别。     

Epidemiology and control of Menangle virus in pigs

OBJECTIVE: To describe the epidemiology and eradication of Menangle virus infection in pigs. DESIGN: Field observations and interventions, structured and unstructured serological surveys, prospective and cross-sectional serological studies and laboratory investigations. PROCEDURE: Serum samples were collected from pigs at a 2600-sow intensive piggery in New South Wales that experienced an outbreak of reproductive disease in 1997. Serum samples were also collected from piggeries that received pigs from or supplied pigs to the affected piggery and from other piggeries in Australia. Serum and tissue samples were collected from pigs at piggeries experiencing reproductive disease in New South Wales. Sera and faeces were collected from grey-headed flying foxes (Pteropus poliocephalus) in the region of the affected piggery. Serum samples were tested for neutralising antibodies against Menangle virus. Virus isolation was attempted from faeces. RESULTS: Following the outbreak of reproductive disease, sera from 96% of adult pigs at the affected piggery, including sows that produced affected litters, contained neutralising antibodies against Menangle virus. Neutralising antibodies were also detected in sera from 88% of finisher pigs at two piggeries receiving weaned pigs from the affected piggery. No evidence of Menangle virus infection was found in other piggeries in Australia. In cross-sectional studies at the affected piggery, colostral antibodies were undetectable in most pigs by 14 to 15 weeks of age. By slaughter age or entry to the breeding herd, 95% of pigs developed high antibody titres (> or = 128) against Menangle virus in the virus neutralisation test. Menangle virus was eradicated from the affected piggery following a program of serological testing and segregation. Neutralising antibodies against Menangle virus were also detected in P poliocephalus from two colonies in the vicinity of the affected piggery. Two piggery workers were infected with Menangle virus. There was no evidence of infection in cattle, sheep, birds, rodents, feral cats and a dog at the affected piggery. CONCLUSIONS: Serological evidence of infection with Menangle virus was detected in pigs at a piggery that had experienced reproductive disease, in pigs at two associated piggeries and in fruit bats in the region of the piggery. Two humans were infected. The mode of transmission between pigs is unknown, but spread by faecal or urinary excretion is postulated. This virus can be eradicated by the segregation of pigs into discrete age groups.     

Specificity of polycllonal and monoclonal antibodies for the identification of Xanthomonas campestris pv. campestris

Polyclonal and monoclonal antibodies (PCAs and MCAs), produced to whole cells and flagellar extracts ofXanthomonas campestris pv.campestris (Xcc), respectively, were tested for specificity. In immunofluorescence microscopy (IF) the three PCAs tested, reacted at low dilutions with all Xcc strains, some other xanthomonads and non-xanthomonads. At higher dilutions most cross-reactivity with non-xanthomonad strains disappeared. However, the cross-reactivity with strains ofX. c. pv.vesicatoria (Xcv),X. c. pv.amoraciae (Xca) andX. c. pv.phaseoli var. fuscans (Xcpf) remained.Six MCA-producing cell clones viz. 20H6, 2F4, 18G12, 10C5, 17C12 and 16B5 were selected for specificity tests with an enzyme immunoassay (EIA), IF and a dot-blot immunoassay (DBI). None of the MCAs reacted with all Xcc strains in IF and EIA. In DBI, only MCAs 17C12 and 16B5 reacted with all Xcc strains. All six MCAs tested, cross-reacted in one of either tests with other pathovars ofX. campestris, such as Xcv or Xca. The MCAs were also tested in immunoblotting experiments using total bacterial extracts, cell envelope and flagellar extracts. MCAs 20H6, 2F4, 18G12 and 10C5 reacted with the lipopolysaccharide (LPS) of Xcc. MCAs 16B5 and 17C12 reacted with a 39 kilodalton and a 29 kilodalton protein, respectively.It is concluded that the PCAs and MCAs discussed in this study may be used for routine identification and differentiation of (a group of) Xcc strains. The significance of the cross-reactions with other pathovars ofX. campestris needs to be determined by testing seed lots.     

Two Hungarian isolates of cucumber mosaic virus from sweet pepper (Capsicum annuum) and melon (Cucumis melo): Identification and antiserum preparation

Two Hungarian virus isolates from sweet pepper (K8) and melon (S4) were identified as cucumber mosaic virus (CMV) on the basis of host plant reactions and serology. The isolates were purified and antisera prepared. Homologous antiserum titers in double-diffusion tests were 256 (K8) and 512 (S4). They were serologically closely related to each other and to other CMV isolates. On the basis of symptoms they belong to different symptomatological groups of CMV; this was supported by serological properties. Sedimentation coefficients were c. 93 S, at 2 mg ml^–1. Purified preparations, stained with 2% uranyl acetate, showed spherical particles. In ELISA purified preparations reacted with each other's antisera.Samenvatting Twee hongaarse virusisolaten uit paprika (K8) en meloen (S4) werden geïdentificeerd als komkommermozaïekvirus (CMV) met behulp van toetsplanten en serologie. Beide isolaten werden gezuiverd en er werden antisera tegen bereid. De homologe titers van de antisera in de agar-geldiffusietoets bedroegen 256 (K8) en 512 (S4). K8 en S4 waren serologisch nauw verwant aan elkaar, evenals aan andere CMV-isolaten. Op grond van hun symptomen op toetsplanten behoren ze tot verschillende symptomatologische groepen van CMV. Dit laatste werd gesteund door de serologische eigenschappen. Beide isolaten hebben een sedimentatiecoëfficiënt van ca 93 S, bij een concentratie van 2 mg ml^–1. Gezuiverde preparaten, gekleurd met 2% uranylacetaat, bleken bolvormige deeltjes te bevatten. In ELISA reageerden gezuiverde preparaten van K8 en S4 met elkaars antisera.     

Studies on the Epidemiology of Tropical Theileriosis (Theileria annulata Infection) in Cattle in Central Anatolia,Turkey

An epidemiological survey for Theileria annulata infection was conducted in 12 selected villages around Ankara in Central Anatolia, Turkey, during the period April 1990 to January 1993. During the survey, 198 cattle of 30 local breeds, 84 Holstein-Friesian×local breeds and 84 Holstein-Friesian breed were examined for antibodies to T. annulata and the presence of the vector ticks. Four species of Hyalomma ticks were identified: Hyalomma anatolicum anatolicum, Hyalomma anatolicum excavtum, Hyalomma detritum and Hyalomma marginatum marginatum. Salivary gland staining indicated that infected adult ticks of all four species were present and, therefore, were implicated in the transmission of tropical theileriosis in the field. Generally, the Hyalomma infestation rate was low, with the heaviest infestations occurring on the older animals. Young adults and calves had very low infestation rates. Most ticks seen on cattle were adults, very few nymphs were found. The blood smear and serological examination of the 198 cattle conducted in March, before the start of the first disease season, showed that the prevalence of piroplasmosis was 11.1% (22 out of 198) and the seroprevalence of T. annulata was 10.6% (21 out of 198). Forty-three animals were then excluded from the study because they were seropositive and/or harboured piroplasms. Ninety-two seronegative animals showed piroplasmosis (92 out of 155) and 34 seronegative animals became seropositive for T. annulata (34 out of 155) during the three disease seasons. One animal became clinically ill with tropical theileriosis and required treatment. The incidence of cattle showing piroplasmosis and disease in the total study sample was 50.7% and 0.5% per disease season, respectively. The seroconversion rate of new infection with T. annulata in the total study was 14.3% per animal season. The number of cattle showing piroplasmosis was much greater than the number of seropositive cattle, which may indicate the presence of another species of Theileria. The two different management systems encountered in the study were considered to have influenced the tick infestation levels.     

Response of Chickens to Infection with Newcastle Disease Virus Isolated from a Guinea Fowl

An isolate of Newcastle disease virus obtained from a guinea fowl was characterized as a viscerotropic velogenic strain based upon pathogenicity index studies. Following inoculation of the viral isolate oronasally into 3-week-old chickens, clinical signs appeared after an incubation period of 4–5 days and included dullness, depression, dyspnoea, diarrhoea and leg paralysis. The virus caused a mortality of 56% with haemorrhages at the tip of the glands of the proventriculus and caecal tonsil. Histopathological changes were prominent in the lymphoid organs, being characterized by depletion, degeneration and necrosis of the lymphoid tissues. The brain was the first organ affected, with changes being noticed 3 days after infection. Isolation of virus from various organs was more frequent from 5 to 10 days after infection, but the virus persisted in some of the organs until 21 days after infection. In spite of the high mortality, a good immune response was elicited by the isolate, as was evident from the antibody titre.     

Immune Responses to Intermediate Strain IBD Vaccine at Different Levels of Maternal Antibody in Broiler Chickens

Tropical Animal Health and Production -     

Genomic and Pathogenic Studies on a Glycoprotein E Variant Field Isolate of Bovine Herpesvirus 1

Glycoprotein E-negative (gE–) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability of this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes.     

Bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep

Objective To study the clinical signs following bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep.
Design A clinical and pathological study.
Procedure Twenty Poll Dorset sheep were inoculated with bluetongue virus serotypes 1 or 3, each inoculum having a different passage history. The sheep were examined daily and their clinical appearance and rectal temperatures recorded. Heparinised and non-heparinised blood samples were taken at intervals for virological and serological study. Gross pathological findings were recorded for several sheep at necropsy and tissue samples were collected from three sheep for virological studies.
Results All inoculated sheep developed clinical disease. The clinical signs and gross pathological changes varied considerably but were consistent with damage to the vascular endothelial system. There was a decline in the titres of infectious bluetongue virus and of antigen in tissues collected between 7 and 12 days after infection.
Conclusions The severity of disease was related to the speed of onset and duration of pyrexia and not the development or titre of viraemia. Generally, those animals with sensitive mouths, depression, coronitis, recumbency and reluctance to move were the most debilitated. Whole blood was the most reliable source of infectious virus from acutely and chronically infected and convalescent animals. However, tissue samples particularly spleen, collected from dead or killed animals suffering from either peracute or acute forms of disease were most appropriate for the rapid confirmation of a clinical diagnosis.     

Epidemiological Observations in a Corriedale Flock affected by Brucella ovis

Brucellosis in sheep, caused by Brucella ovis, is primarily a chronic infectious disease of rams with epididymitis as its most characteristic lesion. Six hundred rams from an infected farm were clinically and serologically examined once a year, over a 3-year period. An increase from 2.1% to 6.3% in the prevalence of animals serologically positive to B. ovis occurred over the 3 years. However, the prevalence of rams with lesions in the reproductive tract declined from 14.2% to 6.5% in the third year following one year of strict culling of clinically affected and rams that were serologically positive for B. ovis. Clinical lesions found in the 179 affected rams fell into two main categories: rams with epididymitis and rams with affected lymph nodes. These results suggest that the prevalence of the disease relates mainly to the sexual activity of the animal and not to age in itself. A single cull based on the results of clinical examination and serological test results was unable to decrease the prevalence of B. ovis in an extensive Corriedale sheep flock.