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Effects of ammonia inhalation and acetic acid pretreatment on colonization kinetics of toxigenic Pasteurella multocida within upper respiratory tracts of swine.

Pigs reared in intensive production systems are continuously exposed to ammonia released by the microbial degradation of their excrement. Exposure to this gas has been shown to increase the severity of the disease progressive atrophic rhinitis by facilitating colonization of the pig's upper respiratory tract by Pasteurella multocida. The etiological mechanism responsible for this synergy was investigated by studying the colonization kinetics of P. multocida enhanced by ammonia and comparing them with those evoked by an established disease model. Three-week-old Large White piglets were weaned and allocated to five experimental groups (groups A to E). Pigs in groups A and B were exposed continuously to ammonia at 20 ppm for the first 2 weeks of the study. Pigs in group C were pretreated with 0.5 ml of 1% acetic acid per nostril on days -2 and -1 of the study. On day 0 all the pigs in groups A, C, and D were inoculated with 1.4 x 10(8) toxigenic P. multocida organisms given by the intranasal route. The kinetics of P. multocida colonization were established by testing samples obtained at weekly intervals throughout the study. The study was terminated on day 37, and the extent of turbinate atrophy was determined by using a morphometric index. The results of the study showed that exposure to aerial ammonia for a limited period had a marked effect on the colonization of toxigenic P. multocida in the nasal cavities of pigs, which resulted in the almost total exclusion of commensal flora. In contrast, ammonia had only a limited effect on P. multocida colonization at the tonsil. The exacerbation of P. multocida colonization by ammonia was restricted to the period of ammonia exposure, and the number of P. multocida organisms colonizing the upper respiratory tract declined rapidly upon the cessation of exposure to ammonia. During the exposure period, the ammonia levels in mucus recovered from the nasal cavity and tonsil were found to be 7- and 3.5-fold higher, respectively, than the levels in samples taken from unexposed controls. Acetic acid pretreatment also induced marked colonization of the nasal cavity which, in contrast to that induced by ammonia, persisted throughout the time course of the study. Furthermore, acetic acid pretreatment induced marked but transient colonization of the tonsil. These findings suggest that the synergistic effect of ammonia acts through an etiological mechanism different from that evoked by acetic acid pretreatment. A strong correlation was found between the numbers of P. multocida organisms isolated from the nasal cavity and the severity of clinical lesions, as determined by using a morphometric index. The data presented in the paper highlight the potential importance of ammonia as an exacerbating factor in respiratory disease of intensively reared livestock.

Acetic Acid↗

Genetic organization of Pasteurella multocida cap Loci and development of a multiplex capsular PCR typing system.

Current serotyping methods classify Pasteurella multocida into five capsular serogroups (serogroups A, B, D, E, and F) and 16 somatic serotypes (serotypes 1 to 16). In the present study, we have developed a multiplex PCR assay as a rapid alternative to the conventional capsular serotyping system. The serogroup-specific primers used in this assay were designed following identification, sequence determination, and analysis of the capsular biosynthetic loci of each capsular serogroup. The entire capsular biosynthetic loci of P. multocida A:1 (X-73) and B:2 (M1404) have been cloned and sequenced previously (J. Y. Chung, Y. M. Zhang, and B. Adler, FEMS Microbiol. Lett. 166:289-296, 1998; J. D. Boyce, J. Y. Chung, and B. Adler, Vet. Microbiol. 72:121-134, 2000). Nucleotide sequence analysis of the biosynthetic region (region 2) from each of the remaining three serogroups, serogroups D, E, and F, identified serogroup-specific regions and gave an indication of the capsular polysaccharide composition. The multiplex capsular PCR assay was highly specific, and its results, with the exception of those for some serogroup F strains, correlated well with conventional serotyping results. Sequence analysis of the strains that gave conflicting results confirmed the validity of the multiplex PCR and indicated that these strains were in fact capsular serogroup A. The multiplex PCR will clarify the distinction between closely related serogroups A and F and constitutes a rapid assay for the definitive classification of P. multocida capsular types.

Animals↗

Pasteurella multocida subsp. multocida and P. multocida subsp. septica differentiation by PCR fingerprinting and alpha-glucosidase activity.

Pasteurella multocida is composed of three subspecies that are often differentiated by fermentation of sorbitol and dulcitol. We studied 35 dulcitol-negative P. multocida isolates from infected dog and cat bite wounds, 16 of which yielded weak and/or conflicting fermentation reactions in Andrades sorbitol, thus making it difficult to distinguish between the two dulcitol-negative subspecies of P. multocida, i.e., P. multocida subsp. multocida and P. multocida subsp. septica. All isolates and two control strains were further analyzed using a PCR fingerprinting technique with a single primer (M13 core) and assessed for alpha-glucosidase (alpha-Glu) activity. Although the PCR fingerprint patterns and alpha-Glu activity did not correlate well with the sorbitol fermentation reactions, they did correlate well with each other. All strains identified as P. multocida subsp. septica were positive for alpha-Glu activity and exhibited the group I PCR fingerprint profile. All strains categorized as P. multocida subsp. multocida displayed either the group II or group III PCR fingerprint profile; 9 of 11 of these isolates were alpha-Glu negative. These data suggest that both PCR fingerprinting and alpha-Glu activity provide reliable means for differentiating P. multocida subsp. multocida from P. multocida subsp. septica, particularly in strains that produce weak and/or discrepant sorbitol fermentation reactions.

Animals↗

Typing of Pasteurella multocida isolated from pigs with and without porcine dermatitis and nephropathy syndrome.

Porcine dermatitis and nephropathy syndrome (PDNS) is a sporadic, usually fatal disease of growing and finishing pigs that has been recognized in many pig-producing countries. Pasteurella multocida strains isolated from 15 pigs with PDNS and 51 pigs without PDNS were characterized by capsule and somatic antigen typing, random amplified polymorphic DNA (RAP-D) typing, and restriction analysis of genomic DNA using pulsed-field gel electrophoresis (PFGE). While capsular, somatic, and RAP-D typing did not discriminate PDNS isolates from non-PDNS isolates, all of the isolates from PDNS cases showed an identical ApaI PFGE restriction pattern. This pattern was also found in a high proportion (36%) of P. multocida strains isolated from non-PDNS cases. Isolation of a single variant of P. multocida from tissues of pigs with PDNS warrants further investigation into the possible role of these bacteria in the etiology of the disease.

Animals↗

Septicemia due to Pasteurella pneumotropica: 16S rRNA sequencing for diagnosis confirmation.

Bacteremia due to Pasteurella pneumotropica occurs infrequently. We report a case of septicemia in a 72-year-old woman who had no underlying illness. The microorganism was isolated from 10 blood cultures and identified by conventional and molecular methods. This is the first reported case of P. pneumotropica septicemia in an immunocompetent patient. The history of P. pneumotropica diseases in animals and humans and their varied clinical features are reviewed.

Aged↗

Random amplified polymorphic DNA and amplified fragment length polymorphism analyses of Pasteurella multocida isolates from fatal fowl cholera infections.

Fowl cholera, a disease caused by Pasteurella multocida, continues to be a major problem for the poultry industry. The sources of pathogenic organisms responsible for most sporadic epidemics remain unconfirmed, although attenuated vaccines that retain a low level of virulence have occasionally been implicated in outbreaks of the disease. One of the vaccines most commonly used to prevent fowl cholera is the M-9 strain. In the present study, 61 clinical isolates from turkeys that died of fowl cholera from 1997 to 1999 on 36 Utah farms were analyzed and compared to the M-9 vaccine strain. Genetic analyses of the isolates were done by random amplified polymorphic DNA (RAPD) analysis and amplified fragment length polymorphism (AFLP) fingerprinting. The results of these genetic analyses were correlated with the vaccination status of the flock, isolate serotype, and geographic location. Although both genetic techniques effectively identified similar subtle genomic differences, RAPD analysis provided only 77% of the detail provided by AFLP analysis. While a relationship between genetic profile and serotype was evident, no significant relationship indicating geographic influence was found (P = 0.351). Interestingly, organisms isolated from vaccinated flocks were significantly closer genetically to the M-9 vaccine strain than isolates from unvaccinated birds were (P = 0.020). Statistical analyses revealed that this relationship could not have been determined by serotyping alone (P = 0.320), demonstrating the value of AFLP and RAPD analyses in the characterization of disease-causing strains.

Animals↗

DNA fingerprinting of Pasteurella multocida recovered from avian sources.

Repetitive sequence-based PCR (rep-PCR) and amplified fragment length polymorphism (AFLP) were used to characterize a sample of 43 field isolates and 4 attenuated vaccine strains of Pasteurella multocida recovered from multiple avian species. Both rep-PCR and AFLP assays were rapid and reproducible, with high indices of discrimination. Concordance analyses of rep-PCR and AFLP with somatic serotyping indicate that, in general, somatic serotyping is a poor indicator of genetic relatedness among isolates of P. multocida. In addition, the data provide evidence of host specificity of P. multocida clones. Overall, the results of our study indicate that the rep-PCR and AFLP techniques enable rapid fingerprinting of P. multocida isolates from multiple avian species and enhance the investigation of fowl cholera outbreaks.

Animals↗

Use of single-enzyme amplified fragment length polymorphism for typing Pasteurella multocida subsp. multocida isolates from pigs.

Single-enzyme amplified fragment length polymorphism (SE-AFLP) analyses were used to differentiate 97 isolates of porcine Pasteurella multocida subsp. multocida. The strains, isolated from animals with pneumonia, rhinitis, and septicemia, were classified as capsular types A, D, and F. SE-AFLP showed a discriminatory index of 0.87 and identified 18 different profiles.

Animals↗

Fatal peritonitis caused by Pasteurella multocida capsular type F in calves.

A fatal case of atypical septicemia of pasteurellosis in veal calves is described. The causative organism was identified as a multiresistant Pasteurella multocida capsular type F isolate. The outbreak was characterized by fibrinous peritonitis and mortality, which are hitherto unreported features of P. multocida capsular type F infections.

Animals↗

Coagglutination test for identification of Pasteurella multocida associated with hemorrhagic septicemia.

Two serogroups (B and E) of Pasteurella multocida associated with hemorrhagic septicemia in cattle were differentiated by a coagglutination test with antibody-coated staphylococci. The group antigen was soluble and heat stable. It was not detectable in unencapsulated cells or their heated extracts. Adsorbed onto human type O erythrocytes, it could be demonstrated by an indirect hemagglutination test with specific antiserum or by coagglutination with specific antibody-coated staphylococci. Antibody-coated staphylococci could detect the soluble group antigen in the plasma and liver extract of mice experimentally infected with P. multocida. The group antigen did not affix to mouse erythrocytes in vivo.

Agglutination Tests↗

Pasteurella haemolytica var. ureae from human sputum.

A new variety of Pasteurella, named by Henriksen and Jyssum (1960, 1961) Past. haemolytica var. ureae, has been isolated from the sputum of 17 patients, most of them elderly, suffering from chronic bronchitis or bronchiectasis. It has also been found in the normal respiratory tract. Whether the organism aggravates the disease or acts as a mere commensal when growing in the damaged tissue is not yet clear.

Aged↗

Waterhouse-Friderichsen syndrome complicating primary biliary sepsis due to Pasteurella multocida in a patient with cirrhosis.

Pasteurella multocida is an opportunistic pathogen causing bacteraemia in patients with liver dysfunction. A fulminant case of acute cholecystitis and septicaemia caused by P multocida, complicated by Waterhouse-Friderichsen syndrome without skin haemorrhage, is reported in a previously healthy 64 year old Chinese woman. The patient presented with a six hour history of sudden onset epigastric pain, vomiting, chills, and rigors. A presumptive diagnosis of cholangitis with septicaemic shock was made. Disease progression was rapid and the patient died within eight hours of symptom onset. This case is further proof that skin and mucosal haemorrhages are not an essential feature of Waterhouse-Friderichsen syndrome and this condition should be suspected in all patients presenting with sudden illness and fulminant septicaemia.

Bacteremia↗

Polymicrobial tenosynovitis with Pasteurella multocida and other gram negative bacilli after a Siberian tiger bite.

Mammalian bites present a considerable clinical problem because they are often associated with bacterial infections. Pasteurella multocida is a microorganism that commonly infects both canine and small feline bites. Zoonotic infections developing after large feline bites have been recognised, although their reports are limited. We describe a 35 year old man who was bitten by a Siberian tiger and who developed infectious tenosynovitis secondary to P multocida, Bergeyella (Weeksella) zoohelcum, and Gram negative bacteria most like CDC group EF-4b and comamonas species. The latter three bacteria have not been isolated previously from large feline bite wounds.

Adult↗

Pasteurella multocida meningitis in an adult: case report.

Pasteurella multocida is known to form part of the normal flora in the nasopharynx or gastrointestinal tract in many domestic and wild animals. Most human P multocida infections are soft tissue infections caused by dog or cat bites. Less commonly this bacterium is associated with infections affecting other organ systems of man. A case of fatal P multocida meningitis discovered at the necropsy of a 52 year old man is described. P multocida is an unusual causative agent of meningitis which tends to affect those at the extremes of age.

Fatal Outcome↗

Occupational exposure to animals and antibodies against Pasteurella multocida.

The relation between occupational exposure to cattle and prevalence of antibodies against Pasteurella multocida was evaluated in 680 workers. Three groups of exposed workers in abattoirs and slaughterhouses (S), in industrial breeding (I), and in traditional breeding (T) were compared with control workers not exposed to cattle or chicken (C). The prevalence of antibodies against capsular antigen A determined by indirect haemagglutination was significantly higher in the exposed groups (S: 26.2%; I: 29.0%; T: 32.1%) than in the control group (C: 14.0%). The prevalence of antibodies against capsular antigen D did not differ significantly between the groups. The prevalence of antibodies against one or more somatic antigens 1,2,3,7,8, or 9 was higher in the exposed groups with a significant difference only for group T versus group C (p less than 0.05). There was also a significant relation between antibodies against capsular antigen A and the contacts with pets. This high prevalence of antibodies against P multocida suggests that the infection is frequently subclinical and not only a disease associated with pets but also an occupationally related infection.

Abattoirs↗

Pasteurella multocida meningitis.

Pasteurella multocida appears to be a very rare cause of meningitis. The bacterium has morphological similarities with other causative organisms, particularly Haemophilus influenzae, and its apparent rarity may be due partly to misidentification. This case, only the fourth reported from the United Kingdom, could not be distinuished clinically from the meningitis, owing to other pyogenic bacteria.

Adult↗

Pericardial tamponade caused by Pasteurella multocida infection after a cat bite.

An unusual case of meningitis and pericardial tamponade caused by Pasteurella multocida after a cat bite is reported. The patient was successfully treated by antibiotics and pericardiocentesis and made an uneventful recovery from a life threatening condition. This case illustrates the potential dangers that can arise from a seemingly trivial and commonplace injury.

Animals↗