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1.
A CELL CULTURE VACCINE AGAINST BOVINE EPHEMERAL FEVER   总被引:1,自引:0,他引:1  
SUMMARY A vaccine was prepared from cell culture fluids harvested from the twelfth passage of the 919 strain of bovine ephemeral fever (BEF) virus in Vero cell cultures. Cattle were vaccinated subcutaneously with various combinations of strain 919 virus and adjuvants. Neutralising antibodies were assayed at various times after vaccination and some cattle were challenged by intravenous inoculation with the virulent 417WBC strain of BEF virus. Strain 919 virus of the third and twelfth passage levels in Vero cells produced neither fever, clinical illness nor detectable viraemia in 5 calves inoculated intravenously. Nor could viraemia be detected in 5 heifers receiving vaccine subcutaneously. When the vaccine was administered mixed with aluminium hydroxide adjuvant, the production of neutralising antibodies increased with an increase in the volume of vaccine from 2.5 ml to 10 ml and the response to 2 injections was significantly better than the response to a single injection. The neutralising antibody response was decreased when vaccine was diluted in phosphate buffered saline. The neutralising antibody response following 2 subcutaneous vaccinations with strain 919 virus mixed with aluminium hydroxide adjuvant was higher than that following intravenous inoculation with virulent virus. The vaccine-induced antibodies persisted for at least 12 months, and revaccination at this time led to an increase in the titre of neutralising antibody. Antibodies induced by a single subcutaneous administration of strain 919 virus mixed with Freund's complete adjuvant persisted for at least 40 weeks; those induced by vaccine containing Freund's incomplete adjuvant had virtually disappeared within 16 weeks. All these calves responded to vaccination with aluminium hydroxide-containing vaccine with increases in levels of neutralising antibodies. Of 26 vaccinated calves challenged with virulent BEF virus, 24 remained clinically normal. Two developed brief periods of pyrexia on the seventh day after challenge, but no other clinical signs. One of these calves had a viraemia that was demonstrated only by intravenous inoculation of a susceptible calf. The remaining calf had no detectable viraemia. All of 7 unvaccinated calves developed severe clinical BEF within 5 days of challenge. No disease attributable to the 919 virus occurred in 24 vaccinated pregnant heifers or their newborn calves.  相似文献   

2.
Three newborn calves were inoculated intracerebrally with bovine ephemeral fever virus strain 525. The 2 that lacked detectable neutralising antibody to bovine ephemeral fever vaccine developed fatal encephalitis after 4 and 7 days respectively. The third calf which had a low level of maternal antibody remained healthy and developed antibody that became undetectable after 6 months. Bovine ephemeral fever virus strain 525 was reisolated from the brains of both dead calves by intracerebral inoculation of suckling mice. Homogenates that were prepared from the brains of the calves failed to produce disease or to induce antibody formation in susceptible calves when inoculated intravenously. Strain 525 of BEF virus has been shown to possess a degree of neurovirulence for laboratory animals that has not been reported for other strains (Tzipori and Spradbrow 1974). Although this strain is unable to produce viraemia in calves after I/V inoculation, the present study shows that strain 525 can multiply in the brain tissues of calves and cause death after I/C inoculation.  相似文献   

3.
Bovine ephemeral fever (BEF) virus vaccines, prepared from the brains of suckling mice infected with strain 525 BEF virus, were evaluated in housed cattle and in the field. The virus in lyophilised preparations was stable for 6 months at -50 degrees C. Thirty-four calves, 5 to 18 months old, were used in laboratory vaccination trials. An increase in serum neutralising antibody was detected in 13 of 14 calves initially free of serum antibody, and all 13 failed to develop clinical illness following challenge with virulent BEF virus. Vaccination resulted in no detectable serum antibody increase in 4 calves, 5 months old, with pre-existing antibody of presumed maternal origin. Seven animals, 18 months of age with serum antibody presumed due to previous BEF infection, developed increased antibody titres following vaccination. In 3 animals vaccinated but not challenged, vaccine-induced antibodies decreased to low levels over 5 months. In contrast, the antibody titres following infection with virulent virus in 2 calves were maintained over 5 months. Field trials, involving 236 animals initially free of serum antibody, were conducted on 5 properties near Mackay and 4 properties near Brisbane. Most of 164 animals were vaccinated with a single dose of lyophilised vaccine containing aluminium hydroxide adjuvant. Only 4 animals failed to develop serum antibody and no adverse reactions to vaccination were reported. Natural infection with BEF occurred in 4 herds at Mackay and clinically mild BEF occurred in 3 of 109 vaccinated and 3 of 46 control animals. On the basis of measured serum antibody titres it was assumed that 8 of 53 animals receiving full vaccine volume, 20 of 40 animals receiving half vaccine volume and 18 of 40 control animals became infected with BEF virus. Two dairy herds in Brisbane became naturally infected with virulent BEF virus 7 months after vaccination. Clinical BEF was observed in 8 of 11 control animals and in 3 of 26 animals which received 2 doses of vaccine. Two strains of BEF virus were isolated from unvaccinated animals that developed clinically mild BEF in the field. These strains either failed to infect, or produced subclinical or very mild BEF, when inoculated intravenously into susceptible calves. The anitbody response to natural infection with apparently mild viruses was short-lived, similar to that produced by vaccination.  相似文献   

4.
OBJECTIVE: To determine whether passively acquired antibodies prevent development of a protective immune response to live virus in calves. ANIMALS: 18 calves. PROCEDURES: Calves were caught immediately after birth and tested free of bovine viral diarrhea virus (BVDV) and serum antibodies against BVDV. Within 48 hours, 12 calves were fed colostrum that contained antibodies against BVDV and 6 calves received BVDV antibody free milk replacer. Three milk replacer fed and 6 colostrum fed calves were exposed to virulent BVDV2-1373 at 2 to 5 weeks of life when passively acquired serum antibody titers were high. After serum antibody titers against BVDV had decayed to undetectable concentrations (at 7 to 9 months of age), the 3 remaining milk replacer fed calves, 6 colostrum fed calves previously exposed to BVDV2-1373, and 6 colostrum fed calves that had not been exposed to the virus were inoculated with BVDV2-1373. RESULTS: Passively acquired antibodies prevented clinical disease in inoculated colostrum fed calves at 2 to 5 weeks of life. Serum antibody titers did not increase in these calves following virus inoculation, and serum antibody titers decayed at the same rate as in noninoculated colostrum fed calves. Inoculated colostrum fed calves were still protected from clinical disease after serum antibody titers had decayed to nondetectable concentrations. Same age colostrum fed calves that had not been previously exposed to the virus were not protected. CONCLUSIONS AND CLINICAL RELEVANCE: A protective immune response was mounted in calves with passive immunity, but was not reflected by serum antibodies titers. This finding has implications for evaluating vaccine efficacy and immune status.  相似文献   

5.
Six pregnant heifers that were experimentally infected with bovine ephemeral fever virus produced normal calves. Foetuses of 8 heifers that were immune to bovine ephemeral fever were inoculated with bovine ephemeral fever virus. One was aborted after 36 days, but the cause of abortion could not be determined. One was sacrificed after 28 days. The foetus and its membranes were normal and neither virus nor antibody was demonstrated. The other 6 heifers produced normal calves. One calf, 501, inoculated after 160 days gestation, contained high levels of neutralising antibody in serum before ingestion of colostrum. The others, inoculated at gestational ages of 52 days to 157 days showed no evidence of neutralising antibody in blood samples collected at birth.  相似文献   

6.
Four 3-month-old Jersey calves and three 3-month-old Holstein calves were inoculated with cervid adenovirus and monitored for clinical signs until necropsied between 10 and 42 days postinoculation. The neonatal Jersey calves had received colostrum, and the Holstein calves were colostrum deprived. Preinoculation and postinoculation serum samples were tested for antibodies to the cervid adenovirus, bovine adenovirus type 6, bovine adenovirus type 7, and goat adenovirus type 1. Virus isolation was performed on kidney, nasal secretion, and/or lung homogenates in fetal white-tailed deer lung cells. Negatively stained preparations of feces from Jersey calves were examined weekly using an electron microscope, and weekly blood samples were collected for complete blood counts. Full necropsies were performed on all calves. A complete selection of tissues was evaluated for microscopic changes, and immunohistochemistry was performed on all tissues using a polyclonal antibody to deer adenovirus. No clinical signs were observed in the calves during the study period. Following inoculation, colostrum-deprived calves developed low antibody titers to deer adenovirus, while the Jersey calves that received colostrum did not. Calves that received colostrum had high antibody titers to bovine adenovirus type 7 and goat adenovirus type 1. No consistent gross or microscopic lesions were seen. Adenovirus was not observed in negatively stained preparations of feces. Immunohistochemistry results did not demonstrate virus in all tissues examined microscopically, and virus was not isolated from lungs, nasal secretions, and kidneys.  相似文献   

7.
Nine calves, were inoculated intravenously with the Innisfail strain of encephalomyocarditis (EMC) virus. Apart from a mild fever, no obvious clinical signs were noted. A low titre viraemia was demonstrated in all 5 calves from which blood was collected, and EMC virus was recovered from the myocardium of 3 of 6 calves at 2, 3 and 6 days after inoculation. Virus was not recovered from the central nervous system. No excretion of EMC virus in urine or faeces was detected in 3 calves. Histopathological lesions were present in brain tissue from only 1 calf, destroyed 14 days after inoculation, and in the heart muscle from another calf, destroyed 7 days after inoculation. Macroscopic lesions were not seen in these organs. Both neutralising and haemagglutination-inhibiting antibodies were produced within one week of infection, reached a peak in 3–4 weeks and persisted undiminished until 9 weeks after inoculation. By nitration on Sephadex G 200, it was shown that the early response was due to IgM type antibodies, and these were replaced by IgG antibody. One calf was inoculated intracerebrally with EMC virus. It developed a flaccid posterior paralysis and was destroyed 6 days later. Virus was recovered from the brain and spinal cord, but no significant histopathological lesions were detected in brain or spinal cord from this calf.  相似文献   

8.
The safety of an Aujeszky's disease virus vaccine based on strain 783, a deletion mutant which does not express glycoprotein I and thymidine kinase, was assessed in pigs, calves and sheep. Four-day-old piglets which were inoculated intranasally and intramuscularly with 10(7) plaque forming units (PFU) developed only slight depression and fever. The virus was transmitted to a sentinel piglet. Six weeks after inoculation, the pigs were injected with high doses of corticosteroids in an attempt to reactivate the vaccine virus. The pigs did not shed Aujeszky's disease virus, did not develop a rise in virus neutralising antibody titres and sentinel pigs remained seronegative to Aujeszky's disease virus. Strain 783 was passaged in two series of three- to five-day old piglets, but after the third and fourth passages virus could no longer be recovered. Pregnant sows were inoculated with 10(7) PFU of virus strain 783 around day 35 or on day 85 of pregnancy, and their fetuses and piglets were assayed for Aujeszky's disease virus and antibodies against Aujeszky's disease virus. No evidence was found for transplacental transmission of the virus. Calves and sheep were given 10(7) PFU of virus strain 783 intranasally or intramuscularly; they survived and did not develop clinical signs of Aujeszky's disease. All the sheep and the calves inoculated intramuscularly developed neutralising antibodies to Aujeszky's disease virus.  相似文献   

9.
The transfer of maternal antibodies to Friesian and buffalo calves born of dams vaccinated against rinderpest was through colostrum only. Colostral antibody titers at the time of parturition were higher than the serum titer. Two hours after suckling, a high level of rinderpest neutralizing antibodies was detected in the sera of newborn animals. The half-life of maternal antibodies in buffalo and Friesian calves was found to be approximately 33 and 29 days respectively. By the age of 7-8 months, 60 per cent of buffalo calves and 80 per cent of Friesian calves had no detectable levels of rinderpest neutralizing antibody.  相似文献   

10.
OBJECTIVE: To evaluate the efficacy of an adjuvanted modified-live bovine viral diarrhea virus (BVDV) vaccine against challenge with a virulent type 2 BVDV strain in calves with or without maternal antibodies against the virus. DESIGN: Challenge study. ANIMALS: 23 crossbred dairy calves. PROCEDURES: Calves were fed colostrum containing antibodies against BVDV or colostrum without anti-BVDV antibodies within 6 hours of birth and again 8 to 12 hours after the first feeding. Calves were vaccinated with a commercial modified-live virus combination vaccine or a sham vaccine at approximately 5 weeks of age and challenged with virulent type 2 BVDV 3.5 months after vaccination. Clinical signs of BVDV infection, development of viremia, and variation in WBC counts were recorded for 14 days after challenge exposure. RESULTS: Calves that received colostrum free of anti-BVDV antibodies and were vaccinated with the sham vaccine developed severe disease (4 of the 7 calves died or were euthanatized). Calves that received colostrum free of anti-BVDV antibodies and were vaccinated and calves that received colostrum with anti-BVDV antibodies and were vaccinated developed only mild or no clinical signs of disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the modified-live virus vaccine induced a strong protective immune response in young calves, even when plasma concentrations of maternal antibody were high. In addition, all vaccinated calves were protected against viral shedding, whereas control calves vaccinated with the sham vaccine shed virus for an extended period of time.  相似文献   

11.
Pregnant sows were inoculated with inactivated transmissible gastroenteritis virus and with preparations of virus surface projections and subviral particles derived by detergent treatment of the virus. Neutralising antibody was demonstrated in serum and colostrum from animals that received whole virus or preparations of surface projections whereas subviral particles failed to stimulate neutralising antibody formation. Similar results were obtained with serum from rabbits inoculated with whole virus and structural components. All three preparations stimulated the formation of agglutinating antibodies, as demonstrated by sedimentation analysis and filtration studies with radiolabelled virus.The immunoglobulin classes responsible for neutralising antibody activity in sows inoculated by the intramammary route were examined. In each case where the immunoglobulin class was determined, IgG was found. One sow that received surface projections also had IgA with neutralising activity in her colostrum. In contrast, infection of sows with live whole virus resulted in neutralising antibody of the IgG, IgM and IgA classes.  相似文献   

12.
OBJECTIVE: To determine whether experimental inoculation with a field strain of epizootic hemorrhagic disease virus serotype-2 (EHDV-2) suspected of causing clinical disease in naturally infected cattle would cause clinical disease in calves. ANIMALS: 8 calves. PROCEDURE: A strain of EHDV-2 isolated from a white-tailed deer that died of hemorrhagic disease was passaged twice in deer and used to inoculate 6 calves SC and ID; the other 2 calves were used as controls. Physical examinations, CBC, lymphocyte blastogenesis assays, and coagulation assays were performed; rectal temperature, interferon production, and serum neutralizing antibody responses were measured; and virus isolation was attempted every other day for 21 days after inoculation and then every fourth day for another 30 days. Calves were euthanatized on postinoculation day 51, and necropsy was performed. RESULTS: Calves inoculated with EHDV-2 became infected, as evidenced by development of viremia and seroconversion. However, the virus did not cause detectable clinical disease, clinicopathologic abnormalities, or gross lesions. Viremia was prolonged despite development of a serum neutralizing antibody response. A white-tailed deer inoculated with the same EHDV-2 strain developed clinical signs of epizootic hemorrhagic disease, demonstrating that the inoculum was virulent. CONCLUSION: Calves experimentally infected with EHDV-2 developed viremia and seroconverted but did not develop detectable clinical disease.  相似文献   

13.
Four Merino sheep inoculated intravenously with bovine blood containing ephemeral fever virus showed no clinical signs of ephemeral fever. Two sheep showed a mild haematological response and developed a neutralising antibody which closely paralled that of a steer inoculated at the same time. Leucocytes harvested from these 2 sheep on days 3 and 4 after inoculation with virus reproduced ephemeral fever when inoculated into susceptible steers whilst those harvested on days 1, 2 and 5 did not. Even though this indicates that EFV can multiply in some sheep when they are inoculated intravenously, it cannot be inferred that natural infection occurs.  相似文献   

14.
Profiles of blood cell counts were evaluated for 15 calves from three different farms. These calves showed petechia in the mucous membranes and in the skin and prolonged secondary bleeding after puncture. The clinical course of the disease could be observed in eleven calves. With exception of one case, the blood cell counts indicated a severe anaemia, leukocytopenia and thrombocytopenia. Out of these 15 calves, six calves survived and the other nine calves died or had to be euthanized due to the severity of the disease. Necropsy of these nine calves revealed petechia in the skin, subcutis, muscles, in inner organs and all serous membranes. Pathohistological examination showed a depletion of the bone marrow and lymphatic tissue in eight calves. These findings confirmed the diagnosis of bovine neonatal pancytopenia (BNP) for eight of these nine calves. Bluetongue virus serotype 8 was tested negatively using PCR. Bovine virus diarrhoea virus (BVDV) was negatively tested using immunofluorescence and cell culture and salmonella species were negatively tested in seven dissected calves. A cluster of toxins was negatively tested in one of the dissected calves. All 15 calves had high antibody titres for BVDV. The BVDV-antibody titres from twelve dams with affected calves were positive in six cases and not detectable in the other six cases. In three of the six dams with not detectable BVDV-antibody titres, calves were fed with colostrum of a further dam with high BVDV-antibody titres. In the further three dams without detectable BVDV-antibody titres, we could not ascertain which colostrum has been fed to the calves. BVDV-specific antigen could not be detected in any of the samples from the calves and dams tested. Using the activity of the gamma-glutamyl-transferase, we assumed a sufficient supply with colostrum for the examined calves.The cause for the occurrence of these BNP cases was due to bone marrow depletion.The reason for the bone marrow depletion remained unclear. However, it was obvious that the BNP described here is highly likely caused by colostrum from cows with positive BVDV-antibody titres.  相似文献   

15.
Potential vertical transmission of wild-type bluetongue virus serotype 8 (BTV-8) in cattle was explored in this experiment. We demonstrated transplacental transmission of wild-type BTV-8 in one calf and oral infection with BTV-8 in another calf. Following the experimental BTV-8 infection of seven out of fifteen multi-parous cows eight months in gestation, each newborn calf was tested prior to colostrum intake for transplacental transmission of BTV by RRT-PCR. If transplacental transmission was not established the calves were fed colostrum from infected dams or colostrum from non-infected dams spiked with BTV-8 containing blood. One calf from an infected dam was born RRT-PCR positive and BTV-specific antibody (Abs) negative, BTV was isolated from its blood. It was born with clinical signs resembling bluetongue and lived for two days. Its post-mortem tissue suspensions were RRT-PCR positive. Of the seven calves fed colostrum from infected dams, none became infected. Of the six calves fed colostrum from non-infected dams spiked with infected blood, one calf became PCR-positive at day 8 post-partum (dpp), seroconverted 27 days later, and remained RRT-PCR and Abs positive for the duration of the experiment (i.e., 70 dpp). This work demonstrates that transplacental transmission in late gestation and oral infection of the neonate with wild-type BTV-8 is possible in cattle under experimental conditions.  相似文献   

16.
To investigate and optimise detection of carriers, we vaccinated 15 calves with an inactivated vaccine based on foot-and-mouth disease virus (FMDV) A Turkey strain and challenged them and two further non-vaccinated calves with the homologous virus four weeks later. To determine transmission to a sensitive animal, we put a sentinel calf among the infected cattle from 60 days post-infection until the end of the experiment at 609 days post-infection. Samples were tested for the presence of FMDV, viral genome, specific IgA antibodies, antibodies against FMDV non-structural (NS) proteins or neutralising antibodies. Virus and viral genome was intermittently isolated from probang samples and the number of isolations decreased over time. During the first 100 days significantly more samples were positive by RT-PCR than by virus isolation (VI), whereas, late after infection more samples were positive by virus isolation. All the inoculated cattle developed high titres of neutralising antibodies that remained high during the entire experiment. An IgA antibody response was intermittently detected in the oropharyngeal fluid of 14 of the 17 calves, while all of them developed detectable levels of antibodies to NS proteins of FMDV in serum, which declined slowly beyond 34 days post-infection. Nevertheless, at 609 days after inoculation, 10 cattle (60%) were still positive by NS ELISA. Of the 17 cattle in our experiment, 16 became carriers. Despite frequent reallocation between a different pair of infected cattle no transmission to the sentinel calf occurred. It remained negative in all assays during the entire experiment. The results of this experiment show that the NS ELISA is currently the most sensitive method to detect carriers in a vaccinated cattle population.  相似文献   

17.
Four lambs and 3 calves, seronegative to bluetongue virus (BTV), were inoculated intravenously with a highly plaque-purified strain of BTV Serotype 10. A single calf and lamb served as controls and were inoculated with uninfected cell culture lysate. All BTV-inoculated lambs exhibited mild clinical manifestations of bluetongue, whereas infected calves were asymptomatic. Viremia persisted in BTV-infected lambs for 35-42 days, and for 42-56 days in BTV-infected calves. Neutralizing antibodies were first detected in sera collected at Day 14 post-inoculation (PI) from 2 BTV-infected calves and all 4 infected lambs, and at Day 28 PI in the remaining calf. The appearance of neutralizing antibody in serum did not coincide with clearance of virus from blood; BTV and specific neutralizing antibody coexisted in peripheral blood of infected lambs and calves for as long as 28 days. The sequential development, specificity and intensity of virus protein-specific humoral immune responses of lambs and calves were evaluated by immunoprecipitation of [35S]-labelled proteins in BTV-infected cell lysates by sera collected from inoculated animals at bi-weekly intervals PI. Sera from infected lambs and calves reacted most consistently with BTV structural proteins VP2 and VP7, and nonstructural protein NS2, and less consistently with structural protein VP5, and nonstructural protein NS1. Lambs developed humoral immune responses to individual BTV proteins more rapidly than calves, and one calf had especially weak virus protein-specific humoral immune responses; viremia persisted longer in this calf than any other animal in the study. The clearance of virus from the peripheral blood of BTV-infected lambs and calves is not caused simply by the production of virus-specific neutralizing antibody, however the intensity of humoral immune responses to individual BTV proteins might influence the duration of viremia in different animals.  相似文献   

18.
OBJECTIVE: To assess the effect of maternal cells or cellular components on neonatal immune responses to intracellular pathogens in calves. ANIMALS: 15 Holstein calves. PROCEDURES: Calves were fed whole colostrum, frozen colostrum, or cell-free colostrum within 4 hours after birth. Leukocytes were obtained from calves before feeding colostrum and 1, 2, 7, 14, 21, and 28 days after ingestion. Proliferative responses against bovine viral diarrhea virus (BVDV) and mycobacterial purified protein derivatives were evaluated. Dams received a vaccine containing inactivated BVDV, but were not vaccinated against mycobacterial antigens. RESULTS: All calves had essentially no IgG in circulation at birth, but comparable and substantial concentrations by day 1. Calves that received whole colostrum had enhanced responses to BVDV antigen 1 and 2 days after ingestion of colostrum. In contrast, calves that received frozen colostrum or cell-free colostrum did not respond to BVDV. No differences were identified among the 3 groups in response to mycobacterial antigens. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that transfer of live maternal cells from colostrum to neonatal calves enhanced responses to antigens against which the dams had previously responded (BVDV), but not to antigens to which the dams were na?ve (mycobacterial purified protein derivatives). Results suggested that cell-mediated immune transfer to neonates can be enhanced by maternal vaccination.  相似文献   

19.
Two groups of 6 newborn goat kids were artificially fed colostrum containing antibody to caprine arthritis-encephalitis (CAE) virus, obtained from clinically affected does. Kids in group A were fed the colostrum from birth until 7 days of age, while kids in group B were fed colostrum from 1 to 3 days after birth for 7 days. Kids were fed cow's milk at all other times. Serum antibody resulting from the consumption of colostrum, detected by agar gel immunodiffusion (AGID) tests, lasted for up to 8 weeks in group A, but none was detected in group B. Four kids from each group became infected with CAE virus as demonstrated by the emergence of active immunity and by virus isolation procedures. It appeared that uptake of colostral antibody by group A did not prevent viral transmission, interfere with development of active immunity, or modify the outcome of the CAE virus infection.  相似文献   

20.
Pairs of newborn calves were exposed to bovine leukemia virus (BLV) when they were given their 1st colostrum feeding. Calves that were given 10(6) BLV-infected lymphocytes in colostrum free of BLV-specific antibody became infected. Calves that were fed 10(7), 10(8), or 10(9) infected lymphocytes in colostrum that contained BLV-specific antibody did not become infected. One of 2 calves inoculated intradermally with 250,000 infected lymphocytes was protected by colostral antibody, but the other was not. Colostral antibody titers in the unprotected calf decreased normally until the calf was 4 months old and then increased markedly; this pattern indicates that the presence of colostral antibody may have prolonged the latent period of the BLV infection.  相似文献   

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