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1.
SUMMARY: Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate Pasteurella multocida isolates from outbreaks of fowl cholera on 7 turkey farms in New South Wales. While only a single isolate was available from 5 of the farms, multiple isolates, 4 and 12 respectively, were available from the other 2 farms. The available field evidence suggested that 8 outbreaks had occurred with one farm suffering 2 outbreaks. The isolates obtained were all confirmed as Pasteurella multocida . Biochemical profiles allocated the isolates to 4 groups, 3 being variants of P multocida subsp multocida and the fourth being P multocida subsp septica . REA performed with Hpall established 7 groups. Ribotyping using the Hpall digests probed with the 16S rRNA operon of Haemophilus paragallinarum recognised the same 7 groups as REA. Unlike the biochemical profiles, both REA and ribotyping provided a fine subdivision that identified outbreaks as either related or unrelated. The REA and ribotyping patterns as well as biochemical profiles were stable for all isolates from the outbreaks in which multiple isolates were obtained from either the same bird or from different birds. REA and ribotyping were found to be superior to biotyping methods for the investigation of fowl cholera outbreaks.  相似文献   

2.
Avian strains of Pasteurella multocida were typed by employing restriction endonuclease analysis (REA) and single enzyme-amplified fragment length polymorphism (AFLP) to evaluate their applicability for epidemiological studies of fowl cholera outbreaks. A total of 72 strains isolated from different avian species (chicken, duck, turkey, quail and goose) belonging to various geographical regions of India were characterized. REA using two different enzymes HhaI and HpaII produced 9 and 18 clusters respectively, whereas Single enzyme-AFLP recognized 32 patterns out of 72 strains typed. The study indicated that REA using HpaII is a simple and resource efficient method, however, further typing with more stringent and rapid method like Single enzyme-AFLP, could drastically enhance investigation in epidemiological studies of fowl cholera outbreaks.  相似文献   

3.
Restriction endonuclease analysis (REA) of whole-cell DNA was used to determine possible sources of Pasteurella multocida for each outbreak of fowl cholera occurring in turkey flocks in eight commercial poultry companies in California from October 1988 to September 1989. Over this period, 179 isolates of P. multocida were obtained from dead turkeys in 80 meat and breeder flocks on 43 premises. P. multocida was isolated from wildlife on five premises. Isolates were characterized by subspecies, serotype, presence of plasmid DNA, and REA type. In 52 (65%) flocks, all isolates of P. multocida had the same REA pattern as the M9 live vaccine strain following digestion of DNA with the restriction enzyme SmaI. Field strains of P. multocida were obtained from 27 (34%) flocks, and one flock (1%) yielded both M9 and a field strain of the organism. REA of field strains of P. multocida revealed 17 different SmaI REA types. Based on matching SmaI REA types, potential sources of P. multocida were identified for 15 of the 28 flocks infected with field strains of the organism, and transmission between turkey premises was a possibility in only seven flocks.  相似文献   

4.
Three California turkey premises that had repeated outbreaks of fowl cholera were studied for periods of 2 to 4 years. Using biochemical, serologic, plasmid DNA, and restriction endonuclease analyses of isolates of Pasteurella multocida from turkeys and wildlife on the premises, strains of the organism were found to be enzootic on two of the premises. On the third, a variety of strains of P. multocida were isolated from fowl cholera outbreak flocks.  相似文献   

5.
Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate a total of 38 Pasteurella multocida isolates from four separate outbreaks of pasteurellosis in Australian piggeries. Six isolates were obtained from Outbreak 1, 16 from Outbreak 2 and eight each from outbreaks 3 and 4. Outbreaks 1 and 2 were cases of pneumonic pasteurellosis while outbreaks 3 and 4 involved systemic pasteurellosis. Biochemical characterisation established that a number of different types of P. multocida were present in outbreaks 1 and 3 while outbreaks 2 and 4 were associated with a single type of P. multocida. Outbreaks 1 and 3 yielded isolates of P. multocida that belonged to the subspecies multocida and gallicida, with the subspecies multocida isolates being identified as biovar 3 (6 in total) or 12 (1 in total) and the subspecies gallicida isolates (7 in total) being identified as biovar 8. All 24 isolates from outbreaks 2 and 4 belonged to the subspecies multocida and were all biovar 3. REA and ribotyping showed that, in outbreaks 1 and 3, there were three different types of P. multocida in each outbreak with no common strains between the outbreaks. The molecular methods showed that only a single strain of P. multocida was associated with outbreaks 2 and 4, although the outbreaks were associated with strains that differed in REA profiles but shared a ribotype profile. This study has shown that both, systemic and pneumonic pasteurellosis can be associated with either a single strain or multiple strains of P. multocida. The results also indicate that the molecular typing methods of REA and ribotyping are superior to biochemical characterisation for epidemiological investigation of porcine pasteurellosis.  相似文献   

6.
A total of 36 tonsil swab samples were collected from healthy swine prior to slaughter at the abattoirs in Can tho and Tien giang provinces of Southern Vietnam. The presence of Pasteurella multocida in these samples was detected by the combination of direct cultivation and isolation, mouse inoculation and the polymerase chain reaction (PM-PCR). P. multocida was detected in 16 samples by PCR, with 17 strains ultimately isolated. All samples were negative for serogroup B by HSB-PCR and conventional serotyping, with isolates identified as A:3, D:1 or D:3. In addition, all samples were determined to be negative for the P. multocida toxin (PMT). Characterisation of isolated P. multocida by REP-PCR and biotyping revealed nine distinct REP profiles and seven biotypes among the 17 isolates. Some correlation was seen with P. multocida isolated from a previous Australian outbreak of acute swine pasteurellosis, and those isolated from fowl cholera outbreaks in Vietnamese poultry.  相似文献   

7.
Samples collected from the oropharynx of wild mammals and birds trapped on 36 turkey farms in California were evaluated for the presence of Pasteurella multocida. A total of 966 animals were collected from 18 premises that had experienced an outbreak of fowl cholera within the past 2-8 months; samples were collected from 16 of these 18 premises within 2-8 weeks of outbreak notification and while the infected flock was still present. A total of 939 animals were trapped from an additional 18 premises that had not reported any outbreaks of fowl cholera within at least 4 months, if ever. Forty-eight isolates of P. multocida, of a variety of somatic serotypes, were recovered from 6 species of mammals and 3 species of birds. On only 2 of 7 premises was the somatic serotype of the isolates obtained from wildlife the same as the isolate obtained from tissues of turkeys that had died of fowl cholera on the same premises. Tests for virulence to turkeys were conducted with 31 of the isolates. Seventeen of these isolates caused mortality in turkeys. Wide ranges in mortality rates and median times to death were observed.  相似文献   

8.
From Aug 1985 through July 1986, 720 meat turkey flocks on 160 California premises were monitored and outbreaks of fowl cholera (Pasteurella multocida) were investigated. Data from 43 outbreak (case) flocks were compared with data from 43 nonoutbreak (control) flocks. Outbreak flocks, compared with control flocks, were more likely to be located on premises with higher maximal bird capacity and history of fowl cholera outbreaks. The overall impression was that flocks in larger, newer, more intensively managed premises were at greater risk of fowl cholera outbreaks than were other flocks.  相似文献   

9.
The live, attenuated vaccine strains of Pasteurella multocida have been hypothesized to be responsible for homologous serotype outbreaks of fowl cholera on farms that use the commercial vaccines. We have further hypothesized that the naturally occurring Clemson University (CU) vaccine strain may be transformed to virulence by the acquisition of plasmid DNA. To test this hypothesis, we obtained seven homologous serotype (A:3,4) P. multocida isolates, all plasmid bearing, that were cultured from fowl cholera cases in vaccinated flocks and compared the isolates with the CU reference vaccine by molecular methods. Restriction fragment length polymorphisms (RFLPs) were detected by DNA/DNA hybridization with labeled probes specific for the cya, aroA, and rrn genes of P. multocida. The RFLPs obtained from BglII-digested genomic DNA probed with cya demonstrated no differences among the isolates. Although three isolates probed with aroA showed a RFLP identical to the vaccine strain, five isolates were distinctly different. Isolates probed with rrn grouped into three different restriction patterns that were dissimilar from that of the vaccine strain. Therefore, we have shown that these fowl cholera isolates are different from the CU vaccine strain and that these outbreaks were not vaccine related.  相似文献   

10.
Five hundred twenty isolates of Pasteurella multocida, collected in California from September 1985 to November 1988, were characterized in the laboratory. Characteristics examined included serotype, capsular type, biotype (subspecies), and possession of plasmid DNA. Three hundred thirty-three isolates recovered from turkeys dying from fowl cholera, 88 isolates from liver turkeys in flocks with fowl cholera outbreaks in the recent past, and 99 isolates from wildlife captured on fowl cholera-outbreak and non-outbreak turkey premises were studied in this manner. Characteristics were fairly homogeneous among isolates, especially those obtained from turkeys. The majority of isolates were serotype 3,4, capsular type A, subspecies multocida, and lacked plasmid DNA. Common serotypes of isolates from turkeys and wildlife sampled on the same premises were noted in eight of 13 cases examined.  相似文献   

11.
Repetitive extragenic palindromic (REP)-PCR (polymerase chain reaction), enterobacterial repetitive intergenic consensus (ERIC)-PCR, and single primer PCR assays were employed to characterize 66 strains of Pasteurella multocida serogroup A:1 isolated from avian species belonging to different regions of India. REP-PCR resulted in amplification of REP sequences from the genome which were in the range of approximately 200 to approximately 3000 bp and accounted for a total of 54 distinguishing profiles (D=0.99). ERIC-PCR analysis also generated amplified products in the range of approximately 200 to approximately 3200 bp categorizing strains into a total of 50 different profiles (D=0.98). Amplification of repetitive regions using a microsatellite primer (GTG)(5), resulted in clear distinctive bands ranging from approximately 200 to approximately 2400 bp. Strains were assigned to 43 profiles (D=0.96). No correlation could be drawn between genotypic profiles and avian hosts with their geographical area of origin. Avian strains of P. multocida serogroup A:1 were found to be highly heterogeneous with diverse profiles. REP-PCR was found to be highly discriminatory and simple method for differentiation of phenotypically similar strains. The present study also indicated that PCR based amplification of repetitive regions of P. multocida is a rapid technique with good discrimination and could be employed directly for routine typing of field isolates from fowl cholera outbreaks.  相似文献   

12.
The applicability of conventional and molecular methods for rapid detection and differentiation of Pasteurella multocida serogroup B isolates involved in an outbreak of haemorrhagic septicaemia affecting Indian buffaloes, was studied. Five isolates were obtained and were subjected to phenotypic and genotypic characterization. None of the five isolates could be differentiated on the basis of cultural, biochemical, pathogenicity and antimicrobial sensitivity patterns. Polymerase chain reaction (PCR)-based techniques were found to be specific and sensitive for rapid detection and differentiation of isolates. Repetitive extragenic palindromic (REP-) PCR, enterobacterial repetitive intergenic consensus (ERIC-) PCR and single-primer PCR differentiated all the five isolates into different profiles. All the isolates involved in the outbreak were found to have a genetic profile different from standard P. multocida strain (P52). However, three isolates had similar profiles, whereas each of the remaining two had a different profile. The study indicates the involvement of multiple strains of P. multocida in a single outbreak of haemorrhagic septicaemia in buffaloes. The results also indicate that molecular methods of detection and typing are superior to conventional methods for rapid epidemiological investigations of haemorrhagic septicaemia.  相似文献   

13.
Swabs of the oropharynges of 801 live turkeys (621 meat birds and 180 breeders), collected from 15 flocks that had experienced an outbreak of fowl cholera and from 12 non-outbreak flocks, were screened for the presence of Pasteurella multocida. Turkeys from outbreak flocks were sampled within 2 to 9 weeks of the outbreak. Forty-nine isolates of P. multocida were recovered from turkeys in 11 of the outbreak flocks, and none were recovered from turkeys in non-outbreak flocks. Isolation rates varied from 0 to 72% of turkeys sampled in a flock. Nineteen isolates were tested for virulence by injecting them intravenously into turkeys, and 14 were lethal. Results demonstrated that for purposes of disease control, meat birds in fowl-cholera-outbreak flocks should be considered carriers of potentially virulent P. multocida for the life of the flock.  相似文献   

14.
Six cases of fowl cholera in growing turkeys and 3 in adult breeder chickens of the broiler type as well as one case each of a Pasteurella (P.) multocida-associated disease in ducklings and goslings were described in consideration of own laboratory findings and available informations of the case history. Furthermore a report is given on a treatment strategy successfully used in turkeys with highly acute fowl cholera. All the P. multocida strains isolated culturally could be assigned to the subspecies multocida. In one case Bordetella avium, Salmonella (S.) arizonae and S. hadar were additionally cultured form part of turkeys submitted. P. multocida and Moraxella (Pasteurella) anatipestifer could be determined as the causative agents of the disease of ducklings and goslings. P. multocida strains from turkeys were identified serologically as serovars A:3.4 (3x), F:3.4 (2x) and A:3 (1x); those from the breeder chickens as A:3 (3x); and one each from ducklings and goslings as F:3.4 and -:3. (uncapsulated). No death occurred in turkeys with clinical signs of a highly acute fowl cholera if the treatment of the affected birds was started with an intravenous injection of sulfadimethoxine and continued with a combination of sulfachlorpyridazine (SCP) and trimethoprim (TMP) given in the drinking water for 5 days. However relapse occurred 2-3 days after withdrawal of the drug, although the therapy was clinically highly effective. The recurrence of the disease could be prevented reliably if the turkeys were vaccinated with an effective oil-based bacterin and subsequently treated with the SCP-TMP combination given in drinking water over a 12 day period.  相似文献   

15.
We have used restriction endonuclease analysis (REA) of genomic DNA to classify porcine Pasteurella multocida isolates with similar capsular and somatic serotypes, and to monitor the distribution of isolates from 12 different herds in Quebec. Within herds, P. multocida isolates of similar capsular and somatic serotypes showed similar REA fingerprints. Between herds, some isolates had similar REA fingerprints. However, differences in REA enabled subtyping of many P. multocida isolates with the same antigen types. Our data indicate that REA would enable accurate epidemiological typing of P. multocida in conjunction with classical capsular and somatic typing.  相似文献   

16.
A serotype-specific polymerase chain reaction (PCR) assay was developed for detection and identification of Pasteurella multocida serotype 1, the causative agent of avian cholera in wild waterfowl. Arbitrarily primed PCR was used to detect DNA fragments that distinguish serotype 1 from the other 15 serotypes of P. multocida (with the exception of serotype 14). Oligonucleotide primers were constructed from these sequences, and a PCR assay was optimized and evaluated. PCR reactions consistently resulted in amplification products with reference strains 1 and 14 and all other serotype 1 strains tested, with cell numbers as low as 2.3 cells/ml. No amplification products were produced with other P. multocida serotypes or any other bacterial species tested. To compare the sensitivity and further test the specificity of this PCR assay with traditional culturing and serotyping techniques, tissue samples from 84 Pekin ducks inoculated with field strains of P. multocida and 54 wild lesser snow geese collected during an avian cholera outbreak were provided by other investigators working on avian cholera. PCR was as sensitive (58/64) as routine isolation (52/64) in detecting and identifying P. multocida serotype 1 from the livers of inoculated Pekins that became sick or died from avian cholera. No product was amplified from tissues of 20 other Pekin ducks that received serotypes other than type 1 (serotype 3, 12 x 3, or 10) or 12 control birds. Of the 54 snow geese necropsied and tested for P. multocida, our PCR detected and identified the bacteria from 44 compared with 45 by direct isolation. The serotype-specific PCR we developed was much faster and less labor intensive than traditional culturing and serotyping procedures and could result in diagnosis of serotype 1 pasteurellosis within 24 hr of specimen submission.  相似文献   

17.
The relationship of an increase in fowl cholera outbreaks in turkeys with an increase in environmental temperatures during June, July, August, and September between 1959 and 1992 was analyzed. High environmental temperatures were found to be influential in the development of fowl cholera in turkeys. When the average monthly maximum environmental temperatures for 5 mo of July and 7 mo of August during the 13 yr between 1967 and 1979 were above 30.5 C, there was a significantly (P < 0.05) higher number of fowl cholera outbreaks in turkeys for each month than during the same months when the average maximum temperatures were below 30.5 C. To test the hypothesis that an increase in fowl cholera outbreaks was preceded by an increase in temperature, the pre- and postoutbreak temperatures for 46 selected outbreak clusters occurring between 1959 and 1992 were averaged. Both the average maximum and minimum temperatures for the latter 9 days of the preoutbreak period were highly significantly (P < 0.001) higher than those of the average cluster outbreak day and the following four postoutbreak days. Also, for the nine individual days of the latter pre-outbreak period, the daily average maximum temperature was significantly (P < 0.05) higher for 3 days and partially significantly (P < 0.10) higher for 3 days than that of the average cluster outbreak day, and the daily average minimum temperature was significantly (P < 0.05) higher for 2 days and partially significantly (P < 0.10) higher for 1 day than that for the average cluster outbreak day.  相似文献   

18.
Pasteurella multocida isolated from turkeys during an outbreak of fowl cholera was characterized by serotype and heterogeneity of genes encoding rRNA (ribotype) to investigate the epidemiology of the organism. Isolates were collected between October 1985 and July 1986. The M9 or Clemson University fowl cholera vaccine-like strain was detected in 17% of the flocks with fowl cholera. One particular strain, isolated only from breeder flocks, was recovered from 7 of the 10 breeder flocks examined in this study. Intracompany transmission appeared to be common, implying a failure in biosecurity. Circumstantial evidence indicated that in the field; the incubation period of P multocida in a turkey flock may be between 2 to 7 weeks. Wildlife did not appear to be an important reservoir of P multocida for turkeys during this study period. Ribotyping results tended to discount several of the possible interflock transmissions, as suggested by examination of serotyping results alone; however, serotyping in combination with ribotyping proved helpful in understanding the epidemiology of P multocida in turkeys.  相似文献   

19.
One hundred twenty-nine multiplier breeder turkey flocks on 45 premises in California were monitored for outbreaks of fowl cholera (FC) (Pasteurella multocida) for 1 year (Aug. 1, 1985, through July 31, 1986). Fourteen (11%) flocks on 10 (22%) premises experienced outbreaks. Nine (64%) outbreaks occurred in the fall or winter. FC-outbreak flocks had significantly shorter lay cycles (24.6 weeks vs. 27.9 weeks) and correspondingly lower total egg production per hen (84 eggs vs. 103 eggs) than non-outbreak flocks. A case-control investigation was performed on 11 FC-outbreak (case) flocks, and nine non-outbreak (control) flocks. Case flocks were located statistically closer to other livestock species than were control flocks (0.28 miles vs. 0.68 miles) and were more likely to utilize on-farm disposal of dead birds.  相似文献   

20.
Broiler breeder chickens were exposed to avirulent Pasteurella multocida at 14, 22, and 34 weeks of age either by stick wing 1 to 3 times or subcutaneously 3 times. Fowl pox vaccine was mixed with the first P. multocida exposure in some groups. Exposure did not impair egg production or hatch of fertile eggs. Challenge with pathogenic P. multocida serotype 1 at 68 weeks indicated that exposure to avirulent P. multocida 2 or 3 times provided better protection than 1 exposure. Mixing fowl pox vaccine with the avirulent P. multocida did not reduce immunity to fowl cholera or fowl pox.  相似文献   

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