Sequential development of antigens and toxins of Pasteurella haemolytica serotype 1 grown in cell culture medium.     

A W Confer  J A Durham 《American journal of veterinary research》1992,53(5):646-652

Pasteurella haemolytica was grown in nonsupplemented cell culture medium, or in medium supplemented with bovine serum albumin (BSA) for 24 hours. The production of leukotoxin (LKT) and endotoxin was sequentially evaluated, as were bacterial antigens associated with bacterial cell lysates and culture supernates. Supplementation of medium with BSA had no effect on bacterial growth curves; however, LKT activity was detected earlier and was greater in culture supernates from BSA-supplemented media than from nonsupplemented medium. Leukotoxin antigen (105 kDa) was detected in culture supernates, using a monoclonal antibody, immunoblot analysis, and densitometry. The relative concentrations of LKT antigen were proportional to LKT activity. Endotoxin activity was initially lowest in the culture supernates from nonsupplemented medium, but increased during the incubation period, whereas endotoxin activity in BSA-supplemented culture supernates decreased with time in culture. In culture supernates from nonsupplemented medium, the number of antigenic bands identified by immunoblot analysis with hyperimmune anti-P haemolytica and densitometry was greater than in culture supernates from supplemented media. In bacterial lysates, a 95-kDa antigen was the major antigen detected, using the anti-LKT monoclonal antibody. The concentration of that antigen varied among lysates from nonsupplemented medium and BSA-supplemented media. Using hyperimmune anti-P haemolytica serum, minor differences were seen in the relative quantities of lysate-associated antigens dependent on time in culture and medium used. Among the major antigens seen, differences were most apparent for 150-, 100-, and 87-kDa antigens, whereas differences were not obvious for 42- 40-, and 30-kDa antigens.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

Sequence diversity of the immunogenic outer membrane lipoprotein PlpE from Mannheimia haemolytica serotypes 1, 2, and 6     

Ayalew S  Blackwood ER  Confer AW 《Veterinary microbiology》2006,114(3-4):260-268

Mannheimia haemolytica serotype 1 (S1), S6 and S2 are the most common bacterial isolates found in shipping fever pneumonia in beef cattle. Currently used vaccines against M. haemolytica do not provide complete protection against the disease. Research with M. haemolytica outer membrane proteins (OMPs) has shown that antibodies to one particular OMP from S1, PlpE, may be important in immunity. Recombinant PlpE (rPlpE) is highly immunogenic in cattle, and the acquired immunity markedly enhanced resistance to experimental challenge. We previously demonstrated that the immunodominant epitope (R2) is located between residues 26 and 76 on the N-terminus of PlpE from a reference S1 strain (). This region consists of eight hexapeptide repeats. The potential of this epitope as a vaccine or supplement to commercial vaccines is dependant on its state of conservation amongst isolates of the three serotypes. To determine this, we sequenced plpE genes from 32 isolates. The sequences from S1 and S6 were identical with one exception. Substantial variation was observed among sequences from S2 strains, particularly in the R2 region of the protein. These variations in S2 isolates range from 3 to 28 hexapeptide repeats. Calculated molecular weight of PlpE from S1 and S6 isolates was 37 kDa, where as PlpE from S2 strains ranged from 30 to 50 kDa. These similarities and differences were demonstrated by western blot. Competitive binding assay was used to determine that antibody against rPlpE from S1 binds native PlpE on surfaces of both S1 and S2 cells.  相似文献   

Isolation and antimicrobial susceptibilities of bacterial pathogens from bovine pneumonia: 1994--2002.   总被引：1，自引：0，他引：1  

Ronald D Welsh  Laura B Dye  Mark E Payton  Anthony W Confer 《Journal of veterinary diagnostic investigation》2004,16(5):426-431

Between 1994 and 2002, a total of 390 (46.3%) Mannheimia haemolytica, 292 (34.7%) Pasteurella multocida, and 160 (19.0%) Histophilus somni were isolated at the Oklahoma Animal Disease Diagnostic Laboratory from lungs from 6-18-month-old beef cattle with pneumonia. The ratio of M. haemolytica isolations to P. multocida isolations decreased from 3.1 in 1994 to 0.8 in 2000 while increasing to 1.5 in 2002. Mannheimia haemolytica isolations significantly (P < 0.05) decreased from 62.5% in 1994 to between 30.6% and 50.4% in 1998--2002. Pasteurella multocida isolations significantly (P < 0.05) increased from 20.0% in 1994 to between 28.6% and 47.4% in 1998--2002. Histophilus somni isolations were <19% except in 1998 (40.8%) and 1999 (23%). Antimicrobial susceptibilities for M. haemolytica significantly declined for erythromycin (P = 0.0001), florfenicol (P = 0.0004), spectinomycin (P = 0.0001), and tilmicosin (P = 0.03). For P. multocida, antimicrobial susceptibilities significantly declined for erythromycin (P = 0.0001), florfenicol (P = 0.004), spectinomycin (P = 0.03), sulfachloropyridizine (P = 0.028), tetracycline (P = 0.017), tilmicosin (P = 0.0001), and trimethoprim/sulfamethoxazole (P = 0.0003). Antimicrobial susceptibilities for H. somni were variable for spectinomycin and sulfachloropyridizine, whereas susceptibilities to other antibiotics remained consistently high.  相似文献   

Diagnosis of infectious canine hepatitis virus (CAV-1) infection in puppies with encephalopathy.     

D Caudell  A W Confer  R W Fulton  A Berry  J T Saliki  G M Fent  J W Ritchey 《Journal of veterinary diagnostic investigation》2005,17(1):58-61

Nine weaned Labrador Retriever puppies from a litter of 11 were presented with signs of acute central nervous system (CNS) disease that included ataxia and blindness. All puppies died. Gross examination of tissues from 2 puppies revealed regionally diffuse hemorrhages in the brain stem and swollen hemorrhagic lymph nodes. Light microscopic examination of hematoxylin and eosin-stained tissues showed numerous large, basophilic intranuclear inclusion bodies within CNS vascular endothelium and occasionally in individual hepatocytes. Immunohistochemical staining of the tissue was positive using an antibody against canine adenovirus-1. Virus isolation for infectious canine hepatitis virus was achieved using inoculated cell cultures. Polymerase chain reaction amplification of DNA from cell culture material revealed shared homology with other mammalian adenoviruses.  相似文献   

Transmission of Bovine viral diarrhea virus 1b to susceptible and vaccinated calves by exposure to persistently infected calves          下载免费PDF全文

Robert W. Fulton  Robert E. Briggs  Julia F. Ridpath  Jeremiah T. Saliki  Anthony W. Confer  Mark E. Payton  Glenn C. Duff  D.L. Step    D.A. Walker 《Canadian journal of veterinary research》2005,69(3):161-169

Bovine viral diarrhea virus (BVDV) persistently infected (PI) calves represent significant sources of infection to susceptible cattle. The objectives of this study were to determine if PI calves transmitted infection to vaccinated and unvaccinated calves, to determine if BVDV vaccine strains could be differentiated from the PI field strains by subtyping molecular techniques, and if there were different rates of recovery from peripheral blood leukocytes (PBL) versus serums for acutely infected calves. Calves PI with BVDV1b were placed in pens with nonvaccinated and vaccinated calves for 35 d. Peripheral blood leukocytes, serums, and nasal swabs were collected for viral isolation and serology. In addition, transmission of Bovine herpes virus 1 (BHV-1), Parainfluenza-3 virus (PI-3V), and Bovine respiratory syncytial virus (BRSV) was monitored during the 35 d observation period. Bovine viral diarrhea virus subtype 1b was transmitted to both vaccinated and nonvaccinated calves, including BVDV1b seronegative and seropositive calves, after exposure to PI calves. There was evidence of transmission by viral isolation from PBL, nasal swabs, or both, and seroconversions to BVDV1b. For the unvaccinated calves, 83.2% seroconverted to BVDV1b. The high level of transmission by PI calves is illustrated by seroconversion rates of nonvaccinated calves in individual pens: 70% to 100% seroconversion to the BVDV1b. Bovine viral diarrhea virus was isolated from 45 out of 202 calves in this study. These included BVDV1b in ranch and order buyer (OB) calves, plus BVDV strains identified as vaccinal strains that were in modified live virus (MLV) vaccines given to half the OB calves 3 d prior to the study. The BVDV1b isolates in exposed calves were detected between collection days 7 and 21 after exposure to PI calves. Bovine viral diarrhea virus was recovered more frequently from PBL than serum in acutely infected calves. Bovine viral diarrhea virus was also isolated from the lungs of 2 of 7 calves that were dying with pulmonary lesions. Two of the calves dying with pneumonic lesions in the study had been BVDV1b viremic prior to death. Bovine viral diarrhea virus 1b was isolated from both calves that received the killed or MLV vaccines. There were cytopathic (CP) strains isolated from MLV vaccinated calves during the same time frame as the BVDV1b isolations. These viruses were typed by polymerase chain reaction (PCR) and genetic sequencing, and most CP were confirmed as vaccinal origin. A BVDV2 NCP strain was found in only 1 OB calf, on multiple collections, and the calf seroconverted to BVDV2. This virus was not identical to the BVDV2 CP 296 vaccine strain. The use of subtyping is required to differentiate vaccinal strains from the field strains. This study detected 2 different vaccine strains, the BVDV1b in PI calves and infected contact calves, and a heterologous BVDV2 subtype brought in as an acutely infected calf. The MLV vaccination, with BVDV1a and BVDV2 components, administered 3 d prior to exposure to PI calves did not protect 100% against BVDV1b viremias or nasal shedding. There were other agents associated with the bovine respiratory disease signs and lesions in this study including Mannheimia haemolytica, Mycoplasma spp., PI-3V, BRSV, and BHV-1.  相似文献   

Relationship of vitamin E supplementation and antimicrobial treatment with acute-phase protein responses in cattle affected by naturally acquired respiratory tract disease     

Carter JN  Meredith GL  Montelongo M  Gill DR  Krehbiel CR  Payton ME  Confer AW 《American journal of veterinary research》2002,63(8):1111-1117

OBJECTIVE: To correlate serum concentrations of fibrinogen (Fib), haptoglobin (Hap), serum amyloid-A (SAA), and alpha-1 acid glycoprotein (AGP) with clinical respiratory tract disease and response to treatment in transport-stressed feedlot cattle fed vitamin E-supplemented diets. ANIMALS: 387 heifer calves (mean initial weight, 197 kg). PROCEDURE: Calves purchased from an order buyer were delivered to a feedlot to study the effects of dietary supplementation with 2,000 IU of vitamin E for 0, 7, 14, or 28 days after arrival. Serum or plasma Fib, Hap, SAA, and AGP concentrations were measured on days 0, 7, and 28 after arrival as well as at the time of treatment for respiratory tract disease with antimicrobial drugs and after completion of treatment. RESULTS: Vitamin E supplementation was associated with decreased treatment costs. In cattle that were not recognized as sick or responded positively to 1 antimicrobial treatment, serum Hap concentrations were significantly lower on days 0 and 7 than concentrations for cattle that required > 1 treatment. Serum Hap concentrations and ratios of Hap to SAA on day 0 significantly correlated with the number of antimicrobial treatments required. Serum Hap concentrations at the time of initial treatment were significantly lower for cattle that required only 1 treatment, compared with those that required > 1 treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Serum Hap concentrations are of potential value for use in assessing feedlot cattle that may become ill as a result of respiratory tract disease and for use in monitoring treatment efficacy.  相似文献   

THE HEALING OF SEGMENTAL BONE DEFECTS, INDUCED BY RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN (rhBMP-2): A RADIOGRAPHIC, HISTOLOGICAL, AND BIOMECHANICAL STUDY IN RATS     

AW Yasko  JM Lane  EJ Fellinger  V. Rosen  JM Wozney  EA Wang 《Veterinary surgery : VS》1993,22(1):10-10

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Function and anatomy of plant siRNA pools derived from hairpin transgenes     

Bess L Chau  Kevin AW Lee 《Plant methods》2007,3(1):13-14

Background  

RNA interference results in specific gene silencing by small-interfering RNAs (siRNAs). Synthetic siRNAs provide a powerful tool for manipulating gene expression but high cost suggests that novel siRNA production methods are desirable. Strong evolutionary conservation of siRNA structure suggested that siRNAs will retain cross-species function and that transgenic plants expressing heterologous siRNAs might serve as useful siRNA bioreactors. Here we report a detailed evaluation of the above proposition and present evidence regarding structural features of siRNAs extracted from plants.  相似文献   

Challenge with Bovine viral diarrhea virus by exposure to persistently infected calves: protection by vaccination and negative results of antigen testing in nonvaccinated acutely infected calves          下载免费PDF全文

Robert W. Fulton  Bill J. Johnson  Robert E. Briggs  Julia F. Ridpath  Jeremiah T. Saliki  Anthony W. Confer  Lurinda J. Burge  Douglas L. Step  Derek A. Walker    Mark E. Payton 《Canadian journal of veterinary research》2006,70(2):121-127

Calves persistently infected (PI) with Bovine viral diarrhea virus (BVDV) represent an important source of infection for susceptible cattle. We evaluated vaccine efficacy using calves PI with noncytopathic BVDV2a for the challenge and compared tests to detect BVDV in acutely or transiently infected calves versus PI calves. Vaccination with 2 doses of modified live virus vaccine containing BVDV1a and BVDV2a protected the calves exposed to the PI calves: neither viremia nor nasal shedding occurred. An immunohistochemistry test on formalin-fixed ear notches and an antigen-capture enzyme-linked immunosorbent assay on fresh notches in phosphate-buffered saline did not detect BVDV antigen in any of the acutely or transiently infected calves, whereas both tests had positive results in all the PI calves.  相似文献   

Vets at War.     

AW English 《Australian veterinary journal》2006,84(5):162-162

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