Effect of temperature, relative humidity and medium on the aerosol stability of infectious bovine rhinotracheitis virus.

Aerosols of infectious bovine rhinotracheitis virus were generated with a Devilbiss 40 nebulizer from Eagle's minimum essential medium, nasal secretion from a noninfected calf and nasal secretion from a calf artificially infected with infectious bovine rhinotracheitis virus and aged in a rotating drum at temperatures of 6 degrees C or 32 degrees C and relative humidities of 30% or 90%. The aerosols were sampled at seven minutes after start of spraying, one hour, two hours and three hours with an all glass impinger (AGI-30) and titrated for infectivity in cell cultures. Physical decay was determined by a rhodamine B tracer technique. During spraying (seven minutes from start of spraying), the virus was usually more stable in aerosols of nasal secretion from a noninfected calf and at 90% relative humidity. In nasal secretion from a noninfected calf the virus survived best at 90% relative humidity when the temperature was 6 degrees C and best at 30% relative humidity when the temperature was 32 degrees C. During aging, biological decay was greater at the higher temperature, and at 6 degrees C, the highest decay rates occurred at 30% relative humidity in Eagle's minimum essential medium and at 90% relative humidity in nasal secretion from a noninfected calf. The stability of infectious bovine rhinotracheitis virus infected nasal secretion was not widely different from that in noninfected nasal secretion, although under certain conditions greater survival occurred in the noninfected secretion.     

牛传染性鼻气管炎诊断和防治

牛传染性鼻气管炎又被称为坏死性皮鼻炎,是由牛传染性鼻气管炎病毒感染引发的一种急性呼吸道疾病。不同年龄的牛感染传染性鼻气管炎病毒后表现的临床症状存在一定差异,年龄较小的牛会表现为明显的临床症状,年龄较大的牛呈现隐性感染或持续性感染,患病牛长时间或终生携带病毒,污染周围环境后造成病情反复流行,病情难以控制、难以消灭。牛传染性鼻气管炎造成的死亡率相对较低,但是会严重影响牛群正常生长与采食,需要掌握该类传染性疾病的流行特点,并进行严格诊断,然后构建有效的防控措施,降低发病率。该文分析牛传染性鼻气管炎的流行病学、临床症状、病理变化、诊断方法和防治措施。     

Effect of Incubation Temperature and Serum Content in Agar Overlay on Plaque Production by Foot-and-Mouth Disease Virus

The numbers of plaques produced by foot-and-mouth disease virus in primary cultures of calf kidney cells conditioned to 24, 30, and 37 C were essentially the same if virus was absorbed at 37 C. Adsorption was as effective at 30 as at 37 C in cultures conditioned to the respective temperature, but 24 C was less effective under this condition. However, when the adsorption temperature for cultures conditioned to 37 C was decreased to 30 or 24 C, fewer plaques were produced than in cultures conditioned to and maintained at the lower temperatures during absorption. Comparison of the number of plaques produced in cultures overlaid with nutrient agar containing from 0.5 to 10% bovine serum revealed no difference in relation to serum concentration. However, plaque size was related directly to the concentration of serum in the overlay.     

Effects of crude extracts of various plants on infectious bovine rhinotracheitis virus-plaque production.

Extracts of 28 plants were tested without demonstable antiviral activity in an agar-overlay plaque-reduction antiviral assay system, using infectious bovine rhinotracheitis virus and bovine endocardial cell cultures. Ethanolic extract of Narcissus tazetta L bulb elicited antiviral activity by inhibition of viral plaque formation. Antiviral activity was demonstrated against infectious bovine rhinotracheitis and equine rhinopneumonitis viruses. Narcissus tazetta L bulb did not directly inactivate the virus extracellularly. The extract exhibited only limited toxicity to rapidly multiplying bovine endocardial cells at plaque-inhibitory levels and was not cytoxic to preformed confluent cell monolayers. Narcissus extract did not induce the formation of drug-resistant viral strains.     

Variation of virus replication in primary bovine kidney cell cultures derived from 68 three-day-old calves

Primary kidney cell cultures were prepared from 68 three-day-old calves. Complete monolayers of these cultures were infected separately with viral diarrhea, infectious bovine rhinotracheitis, parainfluenza, and adenovirus 7 viruses. The yield of virus from all infected cultures was calculated by plaque titer assay after 2 to 4 days' incubation. The variation of virus yield was substantial between individual cultures.     

Detection of Infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea Viruses in the Nasal Epithelial Cells by the Direct Immunofluorescence Technique

Nasal epithelial cells were collected by cotton swabs for the diagnosis in experimental and field cases of infectious bovine rhinotracheitis and field cases of bovine viral diarrhea in calves. A portion of the cells was washed twice in phosphate buffered saline and a 25 µL drop was placed on microscope slides. The cells were dried, fixed and stained according to the direct fluorescent antibody technique. Another portion of the same specimen was inoculated onto primary bovine skin cell cultures for virus isolation. In the experimental studies for infectious bovine rhinotracheitis, 29/35 specimens were positive by fluorescent antibody technique and 32/35 by cell culture and in the field cases, 22/119 were positive by fluorescent antibody technique and 19/119 by cell culture. In the field cases of bovine viral diarrhea, 28/69 samples were positive by fluorescent antibody technique and 14/69 by cell culture. When fluorescent antibody technique was performed on inoculated cell cultures a total of 24/69 specimens were positive for bovine viral diarrhea. The sensitivity of fluorescent antibody technique was thus comparable to that of cell culture method for infectious bovine rhinotracheitis and bovine viral diarrhea.     

Nitrogen metabolism of calves inoculated with bovine adenovirus-3 or with infectious bovine rhinotracheitis virus

Beef calves were inoculated with bovine adenovirus-3 or infectious bovine rhinotracheitis virus. After inoculation, plasma fibrinogen increased, serum phosphorus decreased, and nitrogen and phosphorus digestibility decreased compared with preinoculation values. Urinary N excretion increased when calves developed rectal temperatures greater than 39.7 C. Results indicated that clinical infection of calves with infectious bovine rhinotracheitis virus increases urinary N excretion and reduces N and phosphorus balance, and that clinical and subclinical infections with either virus reduce dietary N digestibility.     

Comparison of persistence of seven bovine viruses on bovine embryos following in vitro exposure

The ability of seven cytopathic strains of bovine viruses to adhere to the zona pellucida of six-to-eight day-old bovine embryos were compared. Embryos were exposed to virus by placing them either in virus suspensions or by culturing them on infected bovine turbinate cultures for 18-24 h. After exposure to bovine virus diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBV), bluetongue virus (BTV), pseudorabies virus (PRV), vesicular stomatitis virus (VSV), parainfluenza 3 virus (PI3), or bovine enterovirus virus (BEV), the embryos were tested for virus by culture in bovine turbinate cells and by morphological examination using electron microscopy (EM). A special technique to minimize loss of embryos processed for EM was developed. More embryos had viral particles on the surface of the zona pellucida after exposure to 18-24 hour infected cell cultures than did embryos exposed to viral culture suspensions. The most dramatic finding was that BTV adhered in large numbers to the surface of the zona pellucida of exposed embryos. IBRV, PRV, and VSV comprised an intermediate group, with virions occasionally detected on the surface of exposed embryos after 5 washes. Therefore, extensive washing is required. The PI3 and BEV were easily removed from embryo-exposed virus by washing. BVD was difficult to identify morphologically, making assessment by EM unreliable. There was no evidence that any one of the seven viruses penetrated the intact zona pellucida. Using a micromanipulator, 42 embryos were also directly inoculated through the zona pellucida with +/- 50 picoliters of virus inoculum or medium.(ABSTRACT TRUNCATED AT 250 WORDS)     

牛传染性鼻气管炎病毒内蒙古分离株gG基因的PCR扩增

参考牛传染性鼻气管炎病毒全基因序列(GenBank)设计1对特异性引物,以牛传染性鼻气管炎病毒内蒙古分离株提取的总DNA为模板,运用PCR方法成功地扩增出牛传染性鼻气管炎病毒内蒙古分离株gG基因,并用琼脂糖凝胶电泳检测扩增产物。     

Protection from parainfluenza-3 virus and persistence of infectious bovine rhinotracheitis virus in sheep vaccinated with a modified live IBR-PI-3 vaccine.

Ewes (N = 7) and their lambs (N = 12) were vaccinated with a commercial modified live infectious bovine rhinotracheitis-parainfluenza type 3 virus vaccine. Both the vaccinated ewes and lambs and a group of unvaccinated ewes (N = 8) and their lambs (N = 13) were subsequently challenged with virulent parainfluenza type 3 virus. Although absolute immunity to infection and clinical response was not conferred, the clinical response was less severe in vaccinated lambs. Vaccinated animals also shed parainfluenza type 3 virus in nasal secretions for a shorter time than nonvaccinated animals. Some vaccinated lambs developed a persistent infectious bovine rhinotracheitis virus infection that was recrudesced by treatment with dexamethasone. It was concluded that vaccination was of benefit in reducing the severity of infection with parainfluenza type 3 virus. However, the inclusion of infectious bovine rhinotracheitis virus in a vaccine for sheep respiratory tract disease is highly questionable as it might increase the risk factor associated with vaccination. The consequences of the persistence of infectious bovine rhinotracheitis virus are now known.     

Susceptibility of cell line cultures of bovine kidney origin to infectious bovine rhinotracheitis and parainfluenza-3 viruses

A comparative study was carried out on the susceptibility of primary bovine embryo kidney (PBEK) cell cultures, and that of AUBEK and MDBK cell lines to infectious bovine rhinotracheitis (IBR) and Parainfluenza-3 (PI-3) viruses.

The cytopathic effects induced by the two viruses were rather inconsistent, based on observations of unstained preparations. On the other hand, there was no significant difference between the susceptibility of the PBEK cultures and the cell line cultures to infection with either virus on the basis of the lesions detected in stained preparations, and of the growth curve patterns.

It is concluded that PBEK cell cultures are more sensitive for isolating IBR or PI-3 viruses than are the AUBEK and MDBK cell lines. However, the latter appear to be satisfactory for studies of these two viruses.     

Thermostability of the vaccination strain of infectious bursal disease virus

The vaccination strain of infectious bursal disease virus, multiplied in cultures of chick embryo cells, was very resistant to heat. At a temperature of 56 degrees C the infection titre of the virus (TCID50) decreased by 0.9 log10 within two hours and by 1.2 log10 within five hours, but the virus remained infective still after 24 hours. At a temperature of 37 degrees C, a slight decrease in infection titre was recorded only after two days and a decrease by 1.2 log10 was recorded within ten days. After the 21st day the virus was almost inactivated. At a temperature of about 20 degrees C the infection titre of the virus decreased linearly from the third to the twelfth weeks. The control samples kept at +4 degrees C retained their infectivity for three months and at -20 degrees C even for six months. The discussion deals with the effect of the concentration of protein and magnesium chloride in the medium on the thermostability of infectious bursal disease virus.     

IBRV对牛单核巨噬细胞吞噬鸡红细胞功能的影响

为研究牛传染性鼻气管炎病毒（IBRV）感染对牛单核巨噬细胞吞噬功能的影响,应用淋巴细胞分离液获得牛外周血单个核细胞,通过贴壁培养成为单核巨噬细胞。 IBRV 感染单核巨噬细胞24 h后加入鸡红细胞,测定被感染细胞对鸡红细胞的吞噬率。结果显示,单核巨噬细胞被IBRV 感染后,对鸡红细胞的吞噬率显著降低（P<0.05）。     

Effects of swab materials on infectious bovine rhinotracheitis virus.

The recovery rates of infectious bovine rhinotracheitis virus from swab materials were compared. The adsorptive and elutive properties of cotton, polyester, and calcium alginate wool were examined by direct exposure of infectious bovine rhinotracheitis virus to swab materials in buffered tissue culture medium. Calcium alginate wool was virucidal; this was apparent after 2 hours' exposure. Cotton and polyester swab materials exhibited little virucidal effects. The addition of wooden applicator sticks with the swab materials reduced viral titers further.     

Susceptibility of bovine macrophage and tracheal-ring cultures to bovine viruses.

Cultures of macrophages initiated from peripheral blood monocytes and organ cultures of tracheal rings were tested for their susceptibility to bovine viruses. With several notable exceptions, viruses cytopathogenic for bovine embryonic lung cultures were cytopathogenic for macrophages. Although cowpox virus replicated in macrophages, pseudocowpox did not, and although pseudorabies virus replicated within macrophages, infectious bovine rhinotracheitis and DN-599 herpesviruses did not. Bluetongue virus established an interesting relationship with macrophages. Whereas bluetongue virus was initially cytopathogenic for macrophages, it lost its cytopathogenicity on repeated passage, although it was capable of continued replication in macrophages. When subsequently passaged onto bovine embryonic lung cultures, it regained its cytopathogenicity. Parainfluenza-3, bovine viral diarrhea, and infectious bovine rhinotracheitis viruses readily destroyed ciliary activity in tracheal-ring cultures, as contrasted with the inability of bovine respiratory syncytial virus to destroy ciliary activity, even though bovine respiratory syncytial virus was able to replicate within ciliated epithelial cells of tracheal rings.     

Detection of African malignant catarrhal fever virus antigens in cell cultures by immunofluorescence

A fluorescent antibody (FA) test for antigens of African bovine wildebeest-derived malignant catarrhal fever virus was developed. Serum from one of the few survivors of the experimental disease in steers was used to prepare the conjugate. Both a virulent and an attenuated strain of malignant catarrhal fever virus were used to infect bovine thyroid cell cultures. Cells infected with both strains were readily detected by FA staining as early as 24 h post infection, whereas cytopathic effect could be observed by bright-field microscopy only after days 5 or 6 post infection. Controls consisting of normal bovine thyroid cells or infected cells treated with conjugated normal globulins did not show autofluorescence. The reaction was blocked by treatment of infected cells with homologous positive antisera but not by treatment with normal bovine serum or antisera to foot-and-mouth disease, rinderpest, bovine virus diarrhea, Ibaraki, infectious bovine rhinotracheitis, or bovine herpes mammilitis viruses. Treated with African malgnant catarrhal fever virus conjugate did not react.     

Interferon, Antibody Responses and Protection Induced by an Intranasal Infectious Bovine Rhinotracheitis Vaccine

The interferon and antibody response induced by an intranasal infectious bovine rhinotracheitis vaccine was followed in 22 calves over a nine month period and the ability of these vaccinated calves to withstand challenge with virulent infectious bovine rhinotracheitis virus was assessed.Interferon was detected two to three days post-vaccination and disappeared by the tenth day. Nasal and serum antibodies appeared by day 7 and persisted for nine months.The calves challenged three days postvaccination came down with disease typical of infectious bovine rhinotracheitis, whereas calves challenged three weeks, three months or nine months postvaccination resisted infection.     

Evidence for defective neutrophil function in lungs of calves exposed to infectious bovine rhinotracheitis virus

Calves were exposed to an aerosol of infectious bovine rhinotracheitis (IBR) virus followed five days later by an aerosol of Pasteurella haemolytica. The animals were subjected to bronchoalveolar lavage before IBR and four days after, and again at 0, 4, 24, and 48 hours following Pasteurella haemolytica challenge. The results of these experiments suggest that neutrophil infiltration into the lung, in response to the presence of the bacteria was delayed thereby allowing the bacteria to become established in the lung. Neutrophils in infected animals displayed little random migration in vitro and did not respond to a chemotactic stimulus. It was also found that alveolar macrophages from virus-infected animals were not able to produce neutrophil chemotactic factors. These data suggest that the decrease in neutrophil chemotaxis and the lack of chemotactic factor production by the alveolar macrophage following infection with infectious bovine rhinotracheitis virus may predispose infected cattle to a secondary bacterial infection.     

牛传染性鼻气管炎防治措施

牛传染性鼻气管炎是由疱疹病毒感染引发的一种急性传染性疾病,临床上以呼吸道黏膜出现严重水肿、充血、坏死、形成浅表性溃疡病灶为主要特征。根据发病部位的不同,主要表现为阴道炎、结膜炎、脑膜炎、流产等其他病症,因此该类疾病属于一病因引发的多病症的传染性疾病。临床上牛传染性鼻气管炎只发生于牛,不会发生于其他动物,病毒的传播流行具有专一性。近年,随牛养殖业的不断向前发展,传染性鼻气管炎的发生率呈现逐渐升高趋势,虽然造成的死亡率较低,但是会严重影响牛群正常生长发育,甚至会造成繁殖母牛严重流产,使繁殖母牛的利用年限极大缩短,引发严重的繁殖障碍。该文主要论述牛传染性鼻气管炎的诊断和防治措施。     

四川部分地区牦牛传染性鼻气管炎血清学检测报告

[目的]为了解四川省牦牛传染性鼻气管炎感染的流行情况。[方法]应用酶联免疫吸附试验对在2012年间采自对四川省甘孜藏族自治州和红原县共377份牦牛血清进行牛传染性鼻气管炎病毒抗体检测。[结果]检出阳性血清样品共105份,阳性率为27.9%。[结论]表明四川省牦牛群中均存在牛传染性鼻气管炎病毒的感染。该研究为该地区牦牛传染性鼻气管防治提供依据。