Monoclonal antibody in the identification of Haemophilus somnus

Electrophoretic comparisons of outer membrane proteins of Haemophilus somnus isolates revealed 2 major protein bands (46 and 14 kilodaltons [kD]) common to all isolates tested. A monoclonal antibody raised against H. somnus reacted to the 46-kD band. Coagglutination tests were performed using a monoclonal antibody coagglutination assay. The monoclonal reagent was produced by incubating Cowan strain Staphylococcus aureus suspension, used as a source of crude protein A, with mouse ascitic fluid monoclonal antibody or goat anti-H. somnus hyperimmune serum. Bacteria to be tested were suspended at a concentration of 4.5 x 10(9) cells/ml. The coagglutination test was performed by the addition of 50 microliters of the monoclonal reagent to 50 microliters of the bacterial suspension on a glass plate and manual rotation for 2-3 minutes. The coagglutination assay using Cowan strain Staphylococcus aureus protein A, coupled with the monoclonal antibody, agglutinated 10 different H. somnus isolates. The antibody reagent did not coagglutinate with Actinobacillus suis, A. equuli, Pasteurella haemolytica, P. multocida, or P. pneumotropica under similar test conditions.     

四种“爱丽丝”牌消毒剂对五种细菌的杀菌效果比较

采用定量悬浮试验法,选择大肠杆菌、鼠伤寒沙门氏菌、多杀性巴氏杆菌、金黄色葡萄球菌和无毒炭疽芽胞杆菌等菌株,对四种未知成份（５％浓度）的消毒剂进行了杀菌效果比较。结果表明,Ⅰ号消毒剂对各菌的杀灭效果均很差,Ⅱ号对沙门氏菌和金黄色葡萄球菌的杀灭效果最好,Ⅲ号对多杀性巴氏杆菌的杀灭效果最好,Ⅳ号对大肠杆菌和炭疽杆菌的杀菌效果最好。比较而言,Ⅱ号消毒剂的杀菌效果最好,Ⅲ号、Ⅳ号次之。     

Turkey macrophage and heterophil bactericidal activity against Pasteurella multocida.

Bactericidal activity of turkey macrophages and heterophils was demonstrated in an in vitro colorimetric bactericidal assay. Two vaccine strains and one field isolate of Pasteurella multocida A:3,4 and a single isolate each of Escherichia coli and Staphylococcus aureus were compared for susceptibility to the bactericidal activity of turkey macrophages and heterophils. Only P. multocida A:3,4-strain M-9 (the least virulent strain) was susceptible to macrophage bactericidal activity in the absence of specific immune serum, whereas all three P. multocida A:3,4 organisms were killed when opsonized with specific immune serum. E. coli was susceptible to the bactericidal activity of macrophages, and S. aureus was resistant. All bacteria tested were highly sensitive to the bactericidal activity of intact turkey heterophils, regardless of the opsonin treatment. Electron microscopic findings suggested that heterophils may kill extracellular P. multocida. Only S. aureus and E. coli were killed by lysed heterophils.     

Disinfection of caliciviruses at 20 and 10 degrees C

Five disinfectants, Venno FF super, Venno Vet 1 super, Venno Oxygen, M&Enno-Veterin?r B neu und Neopredisan 135-1, were tested to evaluate their efficacy against caliciviruses at 20 and 10 degrees C. As model test virus served feline calicivirus type F9 (FCV F9). All disinfectants were tested according to Guidelines of the German Veterinary Association (DVG). The investigations were performed in suspension tests and germ carrier tests. The suspension tests were carried out without and with protein load. As protein was used foetal calf serum at the concentration of 40%. Venno FF super showed less protein dependence, however a considerable temperature dependence. This matter can be corrected by increase of concentration on 2%. Venno Vet 1 super was without protein especially effective. The losses on the effectiveness through low temperature and protein load can be annulled also here by increase of concentration. Venno Oxygen was more effective in the comparison to that here named both preparations. The effects of temperature can be corrected by extension of reaction time. The most effective preparation was M&Enno Veterin?r B neu. The disinfection occurred at 20 degrees C with 0.5% solution within 120 min and at 10 degrees C with 1.0% solution within 60 min. The fifth disinfectant Neopredisan was in suspension tests without protein load and carrier tests with gauze at 20 and 10 degrees C relative convincing but in germ carrier tests with poplar wood, no complete disinfection could be achieved within tested concentrations and reaction times.     

Safety, efficacy and cross-protectivity of a live intranasal aerosol haemorrhagic septicaemia vaccine

The safety, efficacy and cross-protectivity of a live intranasal aerosol haemorrhagic septicaemia vaccine containing Pasteurella multocida serotype B:3,4 were tested in young cattle and buffaloes in Myanmar, where more than 1.5 million animals had been inoculated with this vaccine between 1989 and 1999. A recommended dose of 2 x 10(7) viable organisms was used for the efficacy test. The administration of 100 times the recommended dose to 50 cattle and 39 buffalo calves was innocuous. Seven months after they were vaccinated, three of three buffaloes were protected and 12 months after they were vaccinated, three of four buffaloes were protected against a subcutaneous challenge with serotype B:2 which killed three of three unvaccinated buffaloes. Twelve months after they were vaccinated, eight of eight cattle survived a serotype B:2 challenge, which killed four of four unvaccinated controls. The vaccinated cattle had developed serum antibodies detectable by the passive mouse protection test. Indirect haemagglutination tests on sera taken from cattle 10 days and five weeks after they were vaccinated showed high titres of antibodies. The serum of vaccinated cattle cross-protected passively immunised mice against infection with P. multocida serotypes E:2, F:3,4 and A:3,4.     

Comparative efficacy of several disinfectants in suspension and carrier tests against Haemophilus parasuis serovars 1 and 5

The comparative efficacy of 16 active compounds (including the most commonly used chemical groups) and 10 commercial formulations against Haemophilus parasuis serovars 1 and 5 was studied. These organisms were tested in suspension and carrier tests in the presence and absence of serum as representative of organic matter. Chloramine-T and half of the formulations from commercial sources (most of them including quaternary ammonium compounds) were effective in both in vitro tests, regardless of the presence or absence of organic load. All 26 disinfectants except for an iodophor (0.1% available iodine) resulted in at least 3-log₁₀ reduction in colony-forming units in suspension test, and most of them resulted in the maximal level of detection (>6-log₁₀ reduction). On the other hand, disinfectants were not as effective in carrier test as in suspension test, and the presence of serum considerably reduced the activities of most of the compounds tested, especially in carrier test. These results suggest the importance of selecting suitable disinfection for routine use on surfaces contaminated with H. parasuis, particularly when organic matter is present. Chloramine-T and formulations 2 and 7–10 are recommended for a complete inactivation of H. parasuis in swine herds.     

三氯异氰尿酸高效消毒片消毒试验

用三氯异氰尿酸高效消毒片对部分细菌、新城疫病毒进行定量和定性消毒试验及现场表面自然菌消毒试验，结果0．05％的三氯异氰尿酸溶液对金黄色葡萄球菌ATCC6538、大肠杆菌8099、链球菌C55100、多杀性巴氏杆菌C48．1、新城疫病毒作用5min，杀菌率、病毒灭活率均为100％；对现场表面消毒，0．4％的三氯异氰尿酸溶液对自然菌作用10min，0．2％作用20min，其杀菌率均可达到99．9％以上。     

Ceftiofur sodium, a broad-spectrum cephalosporin: evaluation in vitro and in vivo in mice

Ceftiofur sodium, a broad-spectrum beta-lactamase-resistant cephalosporin, was evaluated in vitro and in vivo in mice. Ceftiofur is the sodium salt of (6R, 7R)-7[( 2-amino-4-thiazolyl)-Z- (methoxyimino)acetyl]amino)-3-[( (2-furanylcarbonyl)thio]methyl)-8-oxo-5- thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylate. Minimal inhibitory concentration values were obtained with 264 strains representing 9 genera and 17 species of bacterial pathogens from cattle, swine, sheep, horses, poultry, dogs, cats, and human beings. Ceftiofur was more active than was ampicillin against all strains tested including beta-lactamase-producing organisms. In mice with systemic infections, ceftiofur was more active than or equivalent to ampicillin, cephalothin, cefamandole, cloxacillin, cefoperazone, or pirlimycin. These protection tests included infections with Escherichia coli, Haemophilus pleuropneumoniae, H somnus, Pasteurella haemolytica, P multocida, Salmonella typhimurium, or Staphylococcus aureus. In infant mice with E coli-induced lethal diarrhea and in mice with S aureus and E coli-induced mastitis, ceftiofur was comparable or more active than was ampicillin.     

In vitro studies with dimethyldithiocarbamate, possible new antimicrobial for use in poultry

Tests were conducted to determine the in vitro efficacy of the dithiocarbamate analogue, dimethyldithiocarbamate (DmDTC), against selected poultry pathogens. Organisms studied were two bacteria, Pasteurella multocida and Escherichia coli, and a mold, Aspergillus fumigatus. Zone of inhibition and the minimum inhibitory concentration were determined for each organism. DmDTC was effective in vitro against all organisms tested, with A. fumigatus showing greatest overall sensitivity.     

Mannan-binding lectin (MBL) in two chicken breeds and the correlation with experimental Pasteurella multocida infection

The present study is the first demonstration of an association of the genetic serum Mannan-binding lectin (MBL) concentration with bacterial infections in chickens. The genetic serum MBL concentration was determined in two chicken breeds, and the association with the specific Pasteurella multocida humoral immune response during an experimental infection was examined. Furthermore, we examined the association of the genetic serum MBL concentration with systemic infection. The chickens with systemic infection had a statistically significant lower mean serum MBL concentration than the rest of the chickens, suggesting that MBL plays an important role against P. multocida. A statistically significant negative correlation was found between the specific antibody response and the genetic serum MBL concentration for both breeds. This indicates that MBL in chickens is capable of acting as the first line of defence against P. multocida by diminishing the infection before the adaptive immune response takes over.     

10日龄肉鸭混合感染新型鸭肝炎病毒和多杀性巴氏杆菌的病原学研究

对1例疑似鸭肝炎病毒和多杀性巴氏杆菌混合感染的10日龄肉鸭采用常规的病毒、细菌鉴定方法和RT-PCR、PCR方法分别进行病毒、细菌的分离与鉴定。病毒鉴定为新型鸭肝炎病毒,细菌鉴定为荚膜血清A型多杀性巴氏杆菌多杀亚种。细菌对SPF鸡的毒力试验结果显示,分离的巴氏杆菌与强毒标准株C48-1毒力相近,为强毒株。细菌对10日龄肉鸭的致病性回归试验结果表明,一定数量的该株巴氏杆菌可导致10日龄雏鸭的感染死亡。结果表明,该批肉鸭为新型鸭肝炎病毒和A型多杀性巴氏杆菌混合感染。这是国内首例从感染鸭肝炎病毒10日龄雏鸭肝脏中分离到多杀性巴氏杆菌。     

Evaluation of virucidal activity of three commercial disinfectants and formic acid using bovine enterovirus type 1 (ECBO virus), mammalian orthoreovirus type 1 and bovine adenovirus type 1

A modified version of the test method of the Comité Européen de Normalisation (CEN) was developed using formic acid and three commercial disinfectants to evaluate virucidal activity against three non-enveloped viruses, bovine enterovirus type 1 (ECBO virus), mammalian orthoreovirus type 1 and bovine adenovirus type 1 (BAV 1). Determination of the effects of temperature was carried out at 20 and 10 degrees C. All tests with protein load used bovine serum albumin (BSA) and yeast extract. The investigations were performed in suspension tests and in carrier tests using poplar wood virus carriers. The carrier tests showed that ECBO virus could be inactivated at 20 degrees C with 1% formic acid within a 60 min reaction time. For disinfection of ECBO virus at 10 degrees C within 60 min, a 2% concentration of formic acid was necessary. Formic acid was ineffective against reovirus and bovine adenovirus and cannot be recommended as a reference disinfectant. Inactivation of ECBO virus and adenovirus type 1 using a disinfectant containing aldehydes and alcohols could be achieved, but only at room temperature. The disinfection of reovirus type 1 at room temperature with this product was possible without a protein load. This disinfectant exhibited disinfection ability at 10 degrees C at a concentration of more than 2% or with a longer exposure time. A disinfectant containing aldehydes was effective at room temperature but its effect was reduced in the presence of organic matter. Inactivation at 10 degrees C was found only against adenovirus. The fourth disinfectant, which contained peroxiacetic acid, inactivated all test viruses at a concentration of 0.5% within 15 min independent of temperature and protein load.     

Comparison of enzyme-linked immunosorbent assay with dot immunobinding assay for detection of antibodies against Pasteurella multocida in turkeys

A dot immunobinding assay (DIA) was developed to detect antibodies against Pasteurella multocida in turkey serum. Five coating antigens, namely, whole-cell (WC) antigen, sonicated cell lysate (SCL), crude capsular extract (CE), formalin extract (FE), and heat-stable antigen (HSA), were compared by enzyme-linked immunosorbent assay (ELISA) and DIA using reference antisera against P. multocida organisms. WC and SCL antigens showed higher sensitivity, whereas FE and HSA antigens were more specific coating antigens in both assays. The specificity of DIA was greater than ELISA by comparing the P/N ratios of HSA against serum prepared from heterologous serotype of P. multocida. The DIA had also several distinct advantages over the ELISA, which included reduction of the manipulation time and more uniform binding of coating antigens onto the nitrocellulose membranes compared with binding of coating antigens to microtiter plates for ELISA.     

Random amplification of polymorphic DNA and phenotypic typing of Zimbabwean isolates of Pasteurella multocida

Eighty-one isolates presumptively identified as Pasteurella multocida from a variety of diseases in animals in Zimbabwe were subjected to biochemical characterization, capsular typing and RAPD analysis. The majority of isolates (over 80%) were assigned into named taxa and were predominantly P. multocida subsp. multocida and P. multocida subsp. septica, whilst the remainder were unassigned. Serogroup A was predominant among the three capsular types (A, B and D) of P. multocida detected. Three main RAPD clusters and three subclusters were observed among the majority of isolates (93.8%), whilst the remainder was found to be weakly related. Nine different groups of strains with similar RAPD profiles (100% similarity) were also observed. The reference strain of capsular serogroup F clustered with the reference strain of P. multocida subsp. septica, whilst all other serogroups clustered with reference strains of subsp. multocida and gallicida. Notably, serogroups A and D were observed to be closely related to the reference strain of subsp. multocida. The relationship between biotype, capsular type, host origin and disease manifestation was not clear-cut. However, most pig isolates of subsp. multocida clustered together as did most cattle isolates of subsp. multocida. RAPD tended to separate subsp. multocida from septica.     

Pasteurellaceae isolated from tonsillar samples of commercially-reared American bison (Bison bison).

As commercial producers of American bison (Bison bison) become more numerous, concerns relative to bison health management increase. Since loss due to respiratory disease associated with Pasteurella and related Pasteurellaceae is a major concern for cattle producers, a study was conducted to determine what types of Pasteurellaceae are carried by bison to evaluate the potential of pneumonic pasteurellosis in bison herds where management practices are comparable to those used for cattle. Tonsillar biopsies, collected in May (n = 29) and August (n = 25) 1997 from 24- to 30-month-old bison bulls, at the time of slaughter were cultured for Pasteurellaceae. Pasteurella spp. were isolated from all the samples collected in May. These included isolates identified as P. haemolytica, trehalosi, testudinis, and multocida subsp. multocida a and multocida b. Actinobacillus spp. and Haemophilus somnus were also isolated from some samples. Pasteurella spp., haemolytica, trehalosi, and multocida subsp. multocida a, multocida b and septica, plus 2 nonspeciated indole-positive biotypes, U2 and U16, were isolated from the second group of tonsil samples. Most of these organisms, including P. haemolytica, P. multocida subsp., and H. somnus are associated with disease in domestic livestock and should be regarded as potential pathogens for bison, particularly in animals which become stressed by management practices commonly used with cattle such as herding, crowding, and shipping.     

Survey of Actinobacillus (Haemophilus) pleuropneumoniae infection in swine by different methods

Lung and serum samples from pigs that died or were emergency-slaughtered in a pooled, conventional fattening herd were examined to survey Actinobacillus pleuro-pneumoniae infection and to compare the sensitivity of different testing methods. A total of 110 lungs were used for cultural isolation of the agent and direct immunofluorescence (IF) of impression smears. Boiled lung suspensions were tested by coagglutination (Co-A) and agar gel precipitation (AGP). Eighty-seven sera were tested along with lung samples from the same pigs. The lungs yielded a varied bacterial flora most often containing Pasteurella multocida and less frequently Actinomyces (Corynebacterium) pyogenes, E. coli and Salmonella. A. pleuropneumoniae was isolated from 30 lungs: from 22 lungs it grew out in pure culture, from 7 as mixed culture with P. multocida and from 1 as mixed culture with A. pyogenes. The number of positive samples obtained by the different methods was as follows: coagglutination test (with boiled lung suspensions): 63 (57.3%); immunofluorescence: 43 (39.2%); AGP test (with serum): 31 (35.6%); AFP test (with boiled lung suspension): 25 (22.7%). A total of 23 samples (20.7%) were negative by all serological tests and by cultural isolation. Most samples gave positive results by two or more tests while 26 samples only by one test (most often, on 13 occasions, by the Co-A test). The Co-A test detected antigenic components of serotypes that have not been isolated in Hungary so far. This indicates that it is not enough to test one strain from a given lung sample: several colonies must be cultured and serotyped.(ABSTRACT TRUNCATED AT 250 WORDS)     

Apparatus for pasteurising teat cup liners between cows in a herringbone parlour.

In a herringbone milking parlour, teat cup liners were deliberately contaminated in turn with Staphylococcus aureus, Escherichia coli, Streptococcus agalactiae and Sterp uberis. Contamination was achieved by filling the liners with milk that contained 10(6) test organisms per ml. After the clusters had been back-flushed with water at 85 degrees C for five seconds, normal swabbing methods failed to recover any contaminating organisms from the teat liners in 56 tests out of 64. After 10 seconds back-flushing no recoveries were made in the same number of tests. The apparatus developed to effect this back-flushing for a particular herringbone parlour is described, with details of its routine use during milking. For a 100-cow herd, the running cost of such equipment using a five-second back-flush is estimated at no more than 4 pounds per week and, in its present form, would not add more than 10 seconds to the total milking time for each cow. Improvements in design of the apparatus, and in milking techniques arising from the routine use of the device, are also considered.     

猪巴氏杆菌、大肠杆菌二联苗的制备及免疫效果观察

以分离某猪场的多杀性巴氏杆菌、致病性大肠杆菌制备猪巴氏杆菌和大肠杆菌二联灭活苗,同时选择病变明显的病料制备组织灭活苗,探讨它们的免疫效果。结果发现它们的免疫保护率分别为：二联灭活苗为80%以上,组织灭活苗为60%。因此二联苗可试用于控制该猪场巴氏杆菌病的流行。     

Immune defence mechanism against blood-borne Pasteurella multocida in turkeys

Humoral and cellular immune defence factors involved in controlling blood-borne Pasteurella multocida were investigated in turkeys by the passive transfer of immune serum or by the treatment with macrophage-activating agents. The treated and untreated birds were intravenously inoculated with a virulent strain of P multocida, and the viable bacteria in the blood, liver and spleen were counted. In untreated birds, the bacteria were rapidly removed from the blood, and the majority were recovered from the liver and spleen 120 minutes after inoculation. Neither the transfer of immune serum nor the treatment with macrophage-activating agents significantly influenced the clearance rate of bacteria from the blood. The number of bacteria recovered from the liver 120 minutes after inoculation was slightly lower in the birds treated with macrophage-activating agents and significantly lower in those given immune serum than in the untreated birds. None of the treatments, however, significantly changed the number of bacteria recovered from the spleen 120 minutes after inoculation. The results suggest that the phagocytes in the liver, but not in the spleen, play a crucial role in the intravascular defence against P multocida in the presence of specific antibodies.     

Fowl cholera: protection against Pasteurella multocida by ribosome-lipopolysaccharide vaccine

Ribosomal protein from Aspergillus fumigatus substituted for intact ribosomes in potentiating the immunogenicity of Pasteurella multocida lipopolysaccharide. Ribosomal protein behaved as a carrier for the lipopolysaccharide. The basic protein methylated albumin, but not protamine sulfate, substituted for ribosomal protein as a carrier for lipopolysaccharide. Synthetic single- and double-stranded polynucleotides did not function as an adjuvant to potentiate the immunogenicity of lipopolysaccharide or methylated albumin-lipopolysaccharide complexes. Double-stranded polynucleotide (poly A:poly U), added as an adjuvant for methylated albumin-lipopolysaccharide vaccine, produced sera with lowered passive hemagglutination antibodies to lipopolysaccharide, but it did not influence protection against challenge with P. multocida. No differences in protection were observed between different lines of specific-pathogen-free white leghorn chickens given ribosome-lipopolysaccharide vaccine. Humoral protection, demonstrated by passive-protection tests, was induced by ribosome-lipopolysaccharide vaccine. Cell-mediated immunity was not detected by delayed-type hypersensitivity skin test reactions.