Fowl cholera in California multiplier breeder turkeys: 1985-86

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.     

A descriptive study of fowl cholera in California meat turkeys: August 1985-July 1986

One hundred sixty meat-turkey premises in California were monitored for outbreaks of fowl cholera from August 1985 through July 1986. Nearly 27 million turkeys in 720 flocks were at risk during the year. Fifty-three flocks of approximately 3 million turkeys on 34 different premises experienced confirmed fowl cholera outbreaks. The epidemic curve for the year indicated that the majority of outbreaks occurred in the late summer and fall, particularly in October. The incidence of outbreaks during this time was not significantly associated with seasonal variation in the size of the turkey population. The mean flock age at outbreak was 10.8 weeks, with a range of 5-18 weeks.     

Restriction endonuclease analysis of Pasteurella multocida isolates from three California turkey premises.

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.     

Homogeneity of characteristics of Pasteurella multocida isolated from turkeys and wildlife in California, 1985-88

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.     

Transmission of Pasteurella multocida on California turkey premises in 1988-89.

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.     

Causes of mortality in commercial organic layers in Denmark

A longitudinal study investigated the courses of mortality in commercial free-range organic layer flocks in Denmark. In total, 15 organic egg-producing flocks from 11 farms were randomly selected among 80 farms registered in Denmark. Four farms with confined egg production on deep litter were included for comparison. Flock sizes ranged from 2260 to 5940 layers. The flocks were monitored from introduction to the layer farm until slaughter. Flock mortalities ranged from approximately 2% to 91%, with a mean of 20.8% for organic flocks compared with 7% for confined flocks on deep litter. In total, 4608 layers were submitted for postmortem examination, representing > 40% of all the dead layers in the investigated flocks. Outbreaks of erysipelas (Erysipelothrix rhusiopathiae) and fowl cholera (Pasteurella multocida) were observed in two and three organic flocks, respectively. The mortality rate reached 91% in one organic flock dually affected by erysipelas and fowl cholera. In six organic flocks, outbreaks of blackhead were diagnosed. Concurrent infections of erysipelas and blackhead were diagnosed in one organic flock. Escherichia coli infections in the form of septicemia were identified in all organic flocks. In addition, cannibalism and constipation contributed significantly to the mortality in some organic flocks. In the confined deep litter flocks, E. coli infection, constipation, and cannibalism represented the most common causes of mortality.     

Pasteurella multocida in wild mammals and birds in California: prevalence and virulence for turkeys

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.     

DNA fingerprinting of plasmid-containing serotype A: 3,4 Pasteurella multocida isolated from cases of fowl cholera in chickens and turkeys

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.     

禽霍乱蜂胶灭活疫苗对鸡鸭鹅的现场免疫应用效果

对鸡、鸭、鹅健康群和禽霍乱发病群现场应用禽霍乱蜂胶灭活疫苗，结果表明，该疫苗安全可靠，不影响开产日龄和产蛋；保护率高，４个月内保护率为９８．７％～１００％，６个月内保护率为８５％～９５％，６个月以上保护率为６０％～８０％；对禽霍乱暴发群配合内服药物，可于５～７ｄ内迅速控制和扑灭疫情，且不复发     

Estimate of economic impact of fowl cholera in turkeys in Georgia in 1986

Information gathered from cases of fowl cholera (FC) in commercial turkey flocks through case records, flock records, and telephone and mail surveys was used to estimate disease costs. The cost to the Georgia commercial turkey industry in 1986 from preventive measures, treatment of outbreaks, and production losses from the disease was estimated at $634,545. The cost of FC per kg of live production was estimated to be $0.015.     

Serologic associations between fowl cholera and other diseases.

As part of a case-control study designed to identify fowl cholera risk factors, 2087 blood samples were collected from 71 California meat-turkey flocks. Samples were tested for antibodies to three mycoplasmas and four viruses pathogenic for turkeys. Flocks that had antibodies to Newcastle disease virus and/or Mycoplasma meleagridis had an increased risk of having an outbreak of fowl cholera. This information should prove useful for fowl cholera control programs in meat turkeys.     

Molecular epidemiology of Pasteurella multocida in turkeys.

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.     

Differentiation of field isolates of Pasteurella multocida serotype 3,4 from live vaccine strain by genotypic characterization

Fifty-five serotype 3,4 isolates of Pasteurella multocida, isolated from turkeys dead from fowl cholera, were characterized (fingerprinted) genotypically for comparison with the serotype 3,4 live fowl cholera vaccine principally used in turkeys in California. Twenty-three isolates were obtained from turkeys vaccinated with the M9 live vaccine, and 32 additional isolates were from turkeys not vaccinated for fowl cholera. Methods of characterization included restriction endonuclease analysis of chromosomal DNA and ribotyping, a technique for highlighting restriction site heterogeneity of highly conserved ribosomal RNA genes and associated sequences using a radiolabeled rRNA probe. Eight different genotypes or ribotypes were detected in these isolates by the above methods. Of 23 isolates from M9-vaccinated turkeys flocks, 19 were the same ribotype as M9. Thirty of 32 isolates recovered from unvaccinated turkeys were different ribotypes from M9. The remaining two isolates resembled M9 and were recovered from two different flocks placed in succession on a turkey farm where a flock placed previously had been vaccinated with M9, suggesting interflock transmission. Ribotyping and restriction endonuclease analysis appear to be useful tools to aid in the determination of the role that the live vaccine plays in fowl cholera epidemiology.     

Possible genetic variation in resistance of turkeys to erysipelas and fowl cholera

Natural disease outbreaks of erysipelas and fowl cholera occurred in several lines of turkeys maintained for genetic studies. There were line differences in mortality during both outbreaks, suggesting that there is genetic variation in resistance to these diseases. A line developed by selection for increased egg production had a higher mortality rate from fowl cholera than did the randombred control line from which it was developed. Both the egg line and its control line had a lower mortality rate in the erysipelas outbreak than did a line selected for increased growth rate. Both diseases induced high mortality in a line selected for increased growth.     

Molecular epidemiology of two fowl cholera outbreaks on a free-range chicken layer farm.

Two outbreaks of fowl cholera on a multiage free-range egg farm were investigated. The outbreaks occurred in 1994 and 2002. A total of 22 strains of Pasteurella multocida were available for study, 11 from the 1994 outbreak and 11 from the 2002 outbreak. Lesions typical of acute fowl cholera were seen in the 1994 outbreak, whereas both acute and chronic fowl cholera occurred in the 2002 outbreak. The isolates were examined in an extended phenotypic typing methodology, by a P. multocida-specific polymerase chain reaction (PCR), by the Heddleston somatic serotyping scheme, and by restriction endonuclease analysis (REA) typing using the enzyme HpaII. All 22 strains had the same phenotypic properties, all were confirmed as P. multocida by PCR, all were Heddleston serovar 4, and all had the same REA pattern. The results indicate that these 2 outbreaks were caused by the same clone of P. multocida--despite the 8-year time period between the outbreaks.     

Characterization of avian strains of Pasteurella multocida by restriction endonuclease and amplified fragment length polymorphism

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.     

Pasteurella multocida recovered from live turkeys: prevalence and virulence in turkeys

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.     

Comparison of various antigens and their ability to detect protective antibodies against Pasteurella multocida using enzyme-linked immunosorbent assay

Various antigenic extracts of the CU strain of Pasteurella multocida were prepared to determine their suitability as plate antigens for use in the enzyme-linked immunosorbent assay (ELISA) for the detection of fowl cholera antibodies. Antisera from two separate broiler breeder flocks with known fowl-cholera-vaccination histories were collected just before the birds were challenged with virulent strain X-73 P. multocida. A potassium thiocyanate (KSCN)-extracted antigen, a capsular (CAP) antigen, a lipopolysaccharide-protein antigen, and heat-stable, salt-soluble antigen were all suitable as ELISA plate-coating antigens. Filtered and unfiltered sonicates of the CU strain of P. multocida were also suitable ELISA plate antigens. The results suggested that different plate antigens were detecting different populations of antibodies formed in response to fowl cholera vaccinations. When antibody titers were correlated with survival after challenge, the KSCN and the CAP plate antigens placed more nonsurvivors into low-antibody-titer ranges and more survivors (protected birds) into the high-antibody-titer ranges than the other plate antigens.     

Relationship of increased fowl cholera outbreaks in turkeys with high environmental temperatures.

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.     

Characterisation of Pasteurella multocida isolated from fowl cholera outbreaks on turkey farms

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.