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Use of random amplification of polymorphic DNA in a case of Pasteurella multocida meningitis that occurred following a cat scratch on the head.

We cultured Pasteurella multocida from the cerebrospinal fluid (CSF) of a 4-month-old infant who presented with meningitis. The patient had been scratched on the head by a cat. Culture of the cat's claws also yielded P. multocida. The isolates had identical biochemical patterns. Analysis of both strains by random amplification of polymorphic DNA and comparison of these strains with P. multocida strains isolated from other cats showed that the two strains were identical and completely different from the unrelated isolates. Our patient's meningitis most likely resulted from direct inoculation of P. multocida into the CSF.

Animals↗

CAT III chloramphenicol resistance in Pasteurella haemolytica and Pasteurella multocida isolated from calves.

Chloramphenicol, which had been used extensively for antimicrobial veterinary therapy, was prohibited in Europe in 1994. Soon after it became available, resistance to this drug was detected, generally conferred by plasmids encoding inactivating enzymes, the chloramphenicol acetyltransferases (CAT), in Gram-negative as well as in Gram-positive bacteria. In the last few years, resistance to antibiotics emerged in Pasteurella strains from breeding herds and this evolution was followed by a national surveillance network. Chloramphenicol-resistance was more recently detected in multiresistant strains. We studied 25 strains of Pasteurella, selected for their resistance to chloramphenicol. Production of a CAT was demonstrated in all these strains. PCR amplification indicated that the CAT produced was of type III for 23 of them. In these strains, chloramphenicol-resistance was mediated by plasmids of about 5.1 kb. Southern blots on restriction fragments suggested a high degree of homology between these 5.1 kb plasmids. In the two other strains, production of a CAT type I was demonstrated, and the corresponding genes were either shown on a plasmid of 17 or 5.5 kb.

Animals↗

Plasmid-borne florfenicol resistance in Pasteurella multocida.

OBJECTIVES: A florfenicol-resistant Pasteurella multocida isolate from a calf was investigated for the genetic basis of florfenicol resistance and the location of the resistance gene. METHODS: The P. multocida isolate 381 was investigated for its in vitro susceptibility to antimicrobial agents and its plasmid content. A 10.8 kb florfenicol-chloramphenicol resistance plasmid, designated pCCK381, was identified by transformation into Escherichia coli. The plasmid was mapped with restriction endonucleases, cloned and sequenced completely. RESULTS: Of the antimicrobials tested, plasmid pCCK381 conferred resistance only to chloramphenicol and florfenicol. It showed extended similarity to the 5.1 kb plasmid pDN1 from Dichelobacter nodosus in the part carrying the mobilization and replication genes. An adjacent 3.2 kb segment was highly homologous to the florfenicol resistance gene region of plasmid pMBSF1 from E. coli. In pCCK381, combined resistance to chloramphenicol and florfenicol was based on the presence of a floR gene that showed 97.2-99.7% identity to so far known floR genes. CONCLUSIONS: The results of this study showed that a plasmid-borne floR gene was responsible for chloramphenicol and florfenicol resistance in the bovine respiratory tract pathogen P. multocida. This is, to the best of our knowledge, the first report of a florfenicol resistance gene in a target bacterium.

Animals↗

Variation in resistance to Pasteurella multocida among turkey lines.

Previous research has shown that a line (F) of turkeys selected long-term for increased 16-wk body weight was more susceptible to challenge with washed Pasteurella multocida than a randombred control line (RBC2), the base population of F. A previous study also indicated that the mortality of the F line following challenge with P. multocida was similar to that of sire lines from two of the three major primary breeders. The purpose of the present study was to compare the resistance of the sire line from the third major primary turkey breeder (C) not previously studied with that of the F and RBC2 lines to determine whether there is variation in resistance among the sire lines from three major primary breeders. The sire lines from all three primary breeders were used in the production of commercial turkeys. Body weights of the F line were greater than those of the C line at the time of challenge with P. multocida. Both the C and F lines were heavier than the RBC2 line. The birds were challenged at 6 wk of age with a field isolate of washed P. multocida (1.2 x 10(7) organism per bird of capsular serogroup A and somatic serotype 3,4). Mortality was recorded daily for 14 d. Mortality following challenge with P. multocida was higher in the F line than in the C line, and both large-bodied lines had higher mortality than the RBC2 line. Based on the present results and those published in the literature, there may be variation in resistance among commercial sire lines from the three major primary breeders.

Animal Husbandry↗

Influence of body weight restriction in a body-weight-selected line of turkeys on response to challenge with Pasteurella multocida.

Previous research has shown that a line (F) of turkeys selected long-term for increased 16-wk BW was more susceptible to challenge with washed Pasteurella multocida (PM) than a randombred control line (RBC2), the base population of the F line. Published research indicated that the mortality of the F line following challenge with PM was similar to that of two commercial sire lines. The purpose of the present study was to determine the influence of reducing BW of the F line to that of the RBC2 line by nutrient restriction on resistance to PM. Four challenge trials were conducted over a 2-yr period. The BW of a group of F line birds was restricted to that of the RBC2 line by limiting access to feed from 1 to 6 wk of age. The F line restricted birds and full-fed RBC2 and F line birds were challenged with a field isolate of washed PM (1.2x10(7) organisms/bird of capsular serogroup A and somatic serotype 3, 4) at 6 wk of age. Birds were checked twice daily for 14 d. Resistance to PM was measured by days to death of those that died and percentage mortality. The BW of the restricted group of the F line did not differ from full-fed RBC2 birds for males or females. In males, the restricted F line birds had similar mortality (48.0%) to the full-fed RBC2 line birds (44.3%), and the mortalities in both groups were significantly lower than that observed for the full-fed F line birds (81.3%) following challenge with PM. The mortality following challenge in females did not differ significantly among groups, even though mortality of the full-fed F line birds (64.1%) and restricted F line birds (63.3%) was more than 9% higher than that (54.2%) observed for the full-fed RBC2 line birds. Days to death was not a sensitive indicator of resistance to PM, as no differences among the three groups of birds were observed for either sex.

Animals↗

Effect of feed withdrawal or challenge with Pasteurella multocida on growth, blood metabolites, circulating growth hormone, and insulin-like growth factor-I concentrations in eight-week-old turkeys.

The daily effects of feed withdrawal or a bacterial disease (Pasteurella multocida; PM) challenge was studied in a slow-growing line of turkeys. The following groups (n = 6 birds/group) were sampled for up to 13 d: untreated control (CON), 4-d feed withdrawal followed by refeeding (FAST), a group that succumbed within the first 2 to 3 d after PM challenge (E-DEAD), a group that succumbed 8 to 9 d after PM challenge (L-DEAD), a group that survived the PM challenge (SUR), and a group treated with both PM challenge and 4-d feed withdrawal followed by refeeding (FAST/CHAL). Daily feed intake and BW gains were markedly reduced in the E-DEAD and L-DEAD groups immediately and 3 d after PM challenge, respectively. Feed intake and BW gain between CON and SUR groups of turkeys were not different throughout the trial. The turkeys in the FAST group followed the expected feed withdrawal and refeeding patterns for feed intake and BW loss or gain. The FAST/CHAL turkeys consumed the minimal amount of feed to maintain BW after refeeding. Plasma uric acid sharply increased 1 d prior to death in both E-DEAD and L-DEAD groups of turkeys. Plasma uric acid also increased each consecutive day during fasting in the FAST and FAST/CHAL groups of turkeys. Plasma growth hormone was measured in only the CON and FAST groups and increased from about 40 to 85 ng/mL in the FAST group during fasting but returned to control levels within 1 d of refeeding. Circulating plasma insulin-like growth factor-I (IGF-I) decreased from about 17 to 5 ng/mL in the PM-challenged (E-DEAD, L-DEAD, and FAST/CHAL groups) and FAST groups. The concentration of IGF-I returned to prefeed withdrawal levels within 3 d of refeeding the FAST group of turkeys. It was concluded that 1) turkey poults that were not susceptible to the PM challenge generally maintained physiological functions at control bird levels, 2) susceptible turkey poults generally exhibited depressed feed intake and BW gains, and 3) poults challenged with both feed withdrawal and PM treatment responded differently than poults challenged with either feed withdrawal or challenge with PM. The depletion of energy intake and mobilization of energy stores in susceptible poults might have contributed to the rate at which PM caused the poults to die.

Animals↗

Serum immunoglobulin G and M concentrations did not appear to be associated with resistance to Pasteurella multocida in a large-bodied turkey line and a randombred control population.

Serum IgG and IgM concentrations in a randombred control line (RBC2) of turkeys and its subline (F), selected long term for increased 16-wk BW were compared. Six-week-old poults were challenged with virulent Pasteurella multocida (1.2 x 10(7) bacteria per bird of serogroup A and serotype 3,4). Sera were collected prior to challenge, and concentrations of IgG and IgM in turkey sera were quantitatively estimated with a Sandwich ELISA using the cross-reactive polyclonal antibodies specific for the chicken Ig isotypes. The F-line turkeys had a higher serum IgM concentration than turkeys from the RBC2 line. There was no significant line difference in IgG concentration. The F line had higher mortality and a shorter number of days to death following challenge with P. multocida than did the RBC2 line. No significant correlation coefficient was found between immunoglobulin isotype concentration in serum and days to death following challenge with P. multocida. The present results suggested that prechallenge IgG and IgM concentrations in serum did not appear to be associated with resistance to P. multocida in a large-bodied turkey line and a randombred control population.

Animal Husbandry↗

Effect of selection for increased body weight in turkeys on lymphoid organ weights, phagocytosis, and antibody responses to fowl cholera and Newcastle disease-inactivated vaccines.

The influence of selection was studied for increased 16-wk BW in turkeys on in vivo phagocytic activity, antibody responses to vaccines, and weight of the spleen and bursa of Fabricius. A line (F) of turkeys selected long term for increased 16-wk BW and its corresponding randombred control (RBC2) were compared. Phagocytic activity was evaluated by the carbon clearance assay. Antibody responses to inactivated Newcastle disease virus and Pasteurella multocida vaccines were examined by ELISA. Body weight and relative weights of spleen and bursa of Fabricius of the two lines were also compared. The F line had lower phagocytic activity than the RBC2 line (P < 0.05). In addition, the F line had greater BW, relative weight of spleen, and ratio of spleen to bursa of Fabricius weight (P < 0.01) but had a lower relative weight of bursa of Fabricius at 9 wk of age. However, there were no line differences in the antibody responses to Newcastle disease virus or P. multocida vaccines at 1, 2, 3, 4, 5, or 12 wk after vaccination. Based on the present results, it is suggested that long-term selection for increased 16-wk BW might have resulted in changes in the immune system, as indicated by changes in the relative weights of the spleen and bursa of Fabricius and phagocytic activity. The decreased phagocytic activity in the F line may be partially responsible for increased susceptibility to specific diseases in this line.

Animals↗

Pasteurella multocida bacteremia and tuboovarian abscess.

BACKGROUND: Tuboovarian abscess is commonly associated with sexually transmitted pathogens. We report a tuboovarian abscess caused by a rare pathogen, Pasteurella multocida, which was managed conservatively. CASE: A 50-year-old sexually inactive woman presented with suprapubic pain and fever. Cat scratches were seen on her hand. Ultrasonography showed a 7.9-cm complex cystic adnexal structure. Her fever persisted despite broad-spectrum parenteral antibiotics. After placement of a transvaginal drain, the patient defervesced, and her pain improved. Both blood cultures and cyst aspirates grew Pasteurella multocida. CONCLUSION: Tuboovarian abscess secondary to rare pathogens must be considered in the differential diagnosis of acute febrile pelvic illness in a non-sexually active woman. Minimally invasive drainage procedures may avoid surgery in patients failing initial antibiotic therapy.

Abscess↗

Pasteurella multocida meningitis and cervical spine osteomyelitis in a neonate.

A 20-day-old male infant presented with fever, decreased alertness and quadriparesis as a result of Pasteurella multocida meningitis and C1-2 vertebral osteomyelitis. Although his household contained 2 pet cats, there was no history of bites, scratches or licks. We speculate that colonization of the nasopharynx was followed by contiguous spread to the retropharyngeal soft tissue, cervical vertebrae and meninges.

Anti-Bacterial Agents↗

Population structure and diversity of avian isolates of Pasteurella multocida from Australia.

A total of 110 isolates of Pasteurella multocida from Australian poultry and reference strains for the 16 somatic serovars plus the three subspecies (gallicida, multocida, septica) were analysed to examine their population structure and diversity. The 81 field isolates examined by multilocus enzyme electrophoresis (MLEE) were diverse, being divided into 56 electrophoretic types (ETs), with the 19 reference strains in another 15 ETs. The population was clonal and somatic serotyping was not particularly useful in establishing relationships between isolates. The 71 ETs formed three distinct subclusters (A, B and C) at a genetic distance of 0.36. Biovars tended to be associated with these subclusters: A with biovars 1, 3, 4, 5 and 8 and B with biovars 2, 6, 7, 9 and 10. Ribotyping, performed on all 110 isolates using Hpall, recognized 21 ribotypes forming nine clusters (R1-R9). The isolates in ribotype cluster R1 were almost identical to those in MLEE cluster B. Using both MLEE and ribotyping, the 19 non-Australian reference strains were found to be distributed over the full diversity of the Australian isolates of P. multocida. This study has shown that a range of P. multocida clones are associated with fowl cholera in Australia and that many of the Australian isolates are similar to non-Australian reference strains. Both the MLEE results and the ribotyping data identified a previously unrecognized subset of P. multocida strains.

Animals↗

Genetic diversity of Pasteurella multocida fowl cholera isolates as demonstrated by ribotyping and 16S rRNA and partial atpD sequence comparisons.

The genetic diversity of Pasteurella multocida, the aetiological agent of fowl cholera, was investigated. The strain collection comprised 69 clinical isolates representing a wide spectrum of hosts and geographic origin. The three type strains for the subspecies of P. multocida were also included. Avian isolates of P. multocida subsp. multocida and P. multocida subsp. septica did not represent separate lines by HpaII ribotyping and the two type strains of mammalian origin (porcine and cat bite) seemed to be representative of avian strains of P. multocida subspp. multocida and septica. By ribotyping, all P. multocida subsp. gallicida strains, except one chicken isolate and the type strain, clustered together. This indicated that the bovine type strain was not representative of this subspecies and that most strains of P. multocida subsp. gallicida are genetically related and may be distantly related to other P. multocida isolates, including those of avian origin. By 16S rRNA and atpD sequence comparisons of selected strains, including both P. multocida isolated from birds and mammals and selected distantly related Pasteurella species associated with birds and mammals, it was found that P. multocida is monophyletic. Extended DNA-DNA hybridizations are highly indicated since strains may exist which would connect the existing subspecies at species level. The considerable genetic diversity of P. multocida fowl cholera isolates is probably related to the clonal nature of this organism, resulting in many divergent lines.

Animals↗

Demonstration that Australian Pasteurella multocida isolates from sporadic outbreaks of porcine pneumonia are non-toxigenic (toxA-) and display heterogeneous DNA restriction endonuclease profiles compared with toxigenic isolates from herds with progressive atrophic rhinitis.

Capsular types A and D of Pasteurella multocida cause economic losses in swine because of their association with progressive atrophic rhinitis (PAR) and enzootic pneumonia. There have been no studies comparing whole-cell DNA profiles of isolates associated with these two porcine respiratory diseases. Twenty-two isolates of P. multocida from diseased pigs in different geographic localities within Australia were characterised genotypically by restriction endonuclease analysis (REA) with the enzyme CfoI. Seven of 12 P. multocida isolates from nasal swabs from pigs in herds where PAR was either present or suspected displayed a capsular type D phenotype. These were shown to possess the toxA gene by polymerase chain reaction (PCR) and Southern hybridisation, and further substantiated by production of cytotoxin in vitro. The CfoI profile of one of these seven isolates, which was from the initial outbreak of PAR in Australia (in Western Australia, WA), was identical with profiles of all six other toxigenic isolates from sporadic episodes in New South Wales (NSW). The evidence suggests that the strain involved in the initial outbreak was responsible for the spread of PAR to the eastern states of Australia. Another 10 isolates, representing both capsular types A and D, were isolated exclusively from porcine lung lesions after sporadic outbreaks of enzootic pneumonia in NSW and WA. CfoI restriction endonuclease profiles of these isolates revealed considerable genomic heterogeneity. Furthermore, none of these possessed the toxA gene. This suggests that P. multocida strains with the toxA gene do not have a competitive survival advantage in the lower respiratory tract or that toxin production does not play a role in the pathology of pneumonic lesions, or both. REA with polyacrylamide gel electrophoresis and silver staining was found to be a practical and discriminatory tool for epidemiological tracing of P. multocida outbreaks associated with PAR or pneumonia in pigs.

Animals↗