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Fulltext Ultrastructural changes in endothelial cells of buffaloes following in-vitro exposure to Pasteurella multocida B:2

Puspitasari Y, Salleh A, Zamri-Saad M

BMC Vet Res, 2020 Jun 09;16(1):186.
PMID: 32517749 DOI: 10.1186/s12917-020-02415-2

BACKGROUND: Pasteurella multocida B:2 causes haemorrhagic septicaemia in cattle and buffaloes. However, buffaloes are found to be more susceptible to the infection than cattle. Upon infection, the pathogen rapidly spread from the respiratory tract to the blood circulation within 16-72 h, causing septicaemia. So far, limited study has been conducted to evaluate the response of endothelial cells of buffalo towards P. multocida B:2 and its lipopolysaccharide (LPS). This study aimed to evaluate the ultrastructural changes in the aortic endothelium of buffaloes (BAEC) following exposure to P. multocida B:2 and its endotoxin. The endothelial cells were harvested from the aorta of healthy buffaloes and were prepared as monolayer cell cultures. The cultures were divided into 3 groups before Group 1 was inoculated with 107 cfu/ml of whole cell P. multocida B:2, Group 2 with LPS, which was extracted earlier from 107 cfu/ml of P. multocida B:2 and Group 3 with sterile cell culture medium. The cells were harvested at 0, 6, 12, 18, 24, 36, and 48 h post-inoculation for assessment of cellular changes using transmission electron microscopy.
RESULTS: The BAEC of Groups 1 and 2 demonstrated moderate to severe endothelial lysis, suggestive of acute cellular injury. In general, severity of the ultrastructural changes increased with the time of incubation but no significant difference (p > 0.05) in the severity of the cellular changes between Groups 1 and 2 was observed in the first 18 h. The severity of lesions became significant (p

Matched MeSH terms: Pasteurella multocida/pathogenicity*; Pasteurella multocida/chemistry
Fulltext Reproductive hormonal variations and adenohypophyseal lesions in pre-pubertal buffalo heifers inoculated with Pasteurella multocida type B: 2 and its immunogens

Jesse FF, Ibrahim HH, Abba Y, Chung EL, Marza AD, Mazlan M, et al.

BMC Vet Res, 2017 Apr 05;13(1):88.
PMID: 28381248 DOI: 10.1186/s12917-017-1010-y

BACKGROUND: Hemorrhagic septicemia is a fatal disease of cattle and buffaloes caused by P. multocida. Although the pathogenesis of the bacteria has been well established in literature, there is a paucity of information on the possible role of the bacteria and its immunogens; lipopolysaccharide (LPS) and outer membrane proteins (OMPs) on the reproductive capacity of buffalo heifers.
METHODS: In this study, twenty one healthy prepubertal female buffaloes aged 8 months were divided into seven groups of 3 buffaloes each (G1-G7). Group 1 (G1) served as the negative control group and were inoculated orally with 10 mL sterile Phosphate Buffer Saline (PBS), groups 2 (G2) and 3 (G3) were inoculated orally and subcutaneously with 10 mL of 10(12) colony forming unit (cfu) of P.multocida type B: 2, while groups 4 (G4) and 5 (G5) received 10 mL of bacterial LPS orally and intravenously, respectively. Lastly, groups 6 (G6) and 7 (G7) were orally and subcutaneously inoculated with 10 mL of bacterial OMPs. Whole blood was collected in EDTA vials at stipulated time points (0, 2, 4, 6, 8, 10, 12, 24, 36, 48, 72, 120, 168, 216, 264, 312, 360, 408, 456 and 504 h), while tissue sections of the pituitary glands were collected and transported to the histopathology laboratory in 10% buffered formalin for processing and Hematoxylin and eosin staining. Plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone (PG), estradiol (EST) and gonadotrophin releasing hormone (GnRH) were determined.
RESULTS: The histopathological lesions observed in the pituitary gland included hemorrhage, congestion, inflammatory cell infiltration, hydropic degeneration, necrosis and edema. These changes were higher (p

Matched MeSH terms: Pasteurella multocida/immunology; Pasteurella multocida/pathogenicity*
Fulltext Interaction between Pasteurella multocida B:2 and its derivatives with bovine aortic endothelial cell (BAEC)

Kamal NM, Zamri-Saad M, Masarudin MJ, Othman S

BMC Vet Res, 2017 Jun 19;13(1):186.
PMID: 28629460 DOI: 10.1186/s12917-017-1109-1

BACKGROUND: Pasteurella multocida B:2 causes bovine haemorrhagic septicaemia (HS), leading to rapid fatalities in cattle and buffaloes. An attenuated derivative of P. multocida B:2 GDH7, was previously constructed through mutation of the gdhA gene and proved to be an effective live attenuated vaccine for HS. Currently, only two potential live attenuated vaccine candidates for HS are being reported; P. multocida B:2 GDH7 and P. multocida B:2 JRMT12. This study primarily aims to investigate the potential of P. multocida B:2 GDH7 strain as a delivery vehicle for DNA vaccine for future multivalent applications.
RESULTS: An investigation on the adherence, invasion and intracellular survival of bacterial strains within the bovine aortic endothelial cell line (BAEC) were carried out. The potential vaccine strain, P. multocida B:2 GDH7, was significantly better (p ≤ 0.05) at adhering to and invading BAEC compared to its parent strain and to P. multocida B:2 JRMT12 and survived intracellularly 7 h post treatment, with a steady decline over time. A dual reporter plasmid, pSRGM, which enabled tracking of bacterial movement from the extracellular environment into the intracellular compartment of the mammalian cells, was subsequently transformed into P. multocida B:2 GDH7. Intracellular trafficking of the vaccine strain, P. multocida B:2 GDH7 was subsequently visualized by tracking the reporter proteins via confocal laser scanning microscopy (CLSM).
CONCLUSIONS: The ability of P. multocida B:2 GDH7 to model bactofection represents a possibility for this vaccine strain to be used as a delivery vehicle for DNA vaccine for future multivalent protection in cattle and buffaloes.

Matched MeSH terms: Pasteurella multocida/genetics; Pasteurella multocida/physiology*
Pasteurella multocida bacteraemia with liver abscess

Sia T, Yong E

BMJ Case Rep, 2024 Jan 16;17(1).
PMID: 38232998 DOI: 10.1136/bcr-2023-258386

A previously healthy woman in her mid-70s presented with right upper quadrant abdominal pain, fever, intermittent chills and malaise for 1 week. She was clinically septic with raised inflammatory markers. Her blood culture revealed Pasteurella multocida, which was susceptible to penicillin and amoxicillin-clavulanic acid. CT of liver revealed an abscess of 8.0×7.9×8.5 cm at the left lobe of the liver. However, the abscess was not amenable for surgical or radiological drainage. She was a farmer and had close contact with her pet cats. She was occasionally scratched by her cats when caring for them. The liver abscess resolved completely without drainage after prolonged antimicrobial therapy of 109 days. She commenced on 63 days of intravenous antimicrobials and 46 days of oral amoxicillin-clavulanic acid. This case illustrated P. multocida bacteraemia with a large liver abscess in an immunocompetent adult after non-bite exposure.

Matched MeSH terms: Pasteurella multocida*
Immunogenicity of purified lipopolysaccharide or protein-oligosaccharide conjugates of Pasteurella multocida type 6:B in mice

Muniandy N, Love DN, Mukkur TK

Comp Immunol Microbiol Infect Dis, 1998 Oct;21(4):257-79.
PMID: 9775357

Purified lipopolysaccharide (LPS) of Pasteurella multocida type 6:B, while toxic at higher doses, was protective at lower dose levels against experimentally-induced pasteurellosis in mice. However, the observed protection was abrogated if such LPS was digested with proteinase K prior to use in immunisation. The O-antigen polysaccharide side-chain (OS) of LPS did not appear to contribute to the observed protection as judged by the fact that immunisation of mice with purified OS or OS-protein conjugates, all of which were nontoxic, failed to confer protection against challenge with homologous virulent organisms. This was despite generation of significant levels of OS-specific antibodies, predominantly either of the IgM or IgG isotypes, in immunised mice.

Matched MeSH terms: Pasteurella multocida/immunology*
Improved stability of live attenuated vaccine gdhA derivative Pasteurella multocida B:2 by freeze drying method for use as animal vaccine

Oslan SNH, Halim M, Ramle NA, Saad MZ, Tan JS, Kapri MR, et al.

Cryobiology, 2017 12;79:1-8.
PMID: 29037980 DOI: 10.1016/j.cryobiol.2017.10.004

The efficacy of attenuated strain of gdhA derivative Pasteurella multocida B:2 mutant as a live vaccine to control haemorrhagic septicaemia (HS) disease in cattle and buffaloes has been demonstrated. In order to use P. multocida B:2 mutant as a commercial product, it is essential to optimise its formulation for high viability and stability of the live cells. The effectiveness of freeze-drying process using different protective agent formulations for improving cells viability was explored. Sugar and nitrogen compounds were used as protective agents in freeze-drying and the capability of these compounds in maintaining the viability of mutant P. multocida B:2 during subsequent storage was investigated. A complete loss in viability of freeze-dried mutant P. multocida B:2 was monthly observed until 6-12 months of storage at -30 °C, 4 °C and 27 °C when nitrogen compound or no protective agent was added. Trehalose and sucrose showed significantly high survival rate of 93-95% immediately after freeze-drying and the viability was retained during the subsequent storage at -30 °C and 4 °C. A smooth cell surface without any cell-wall damage was observed for the cells formulated with trehalose under scanning electron micrograph. This study presented a freeze-drying process generating a dried live attenuated vaccine formulation with high stability for commercial applications.

Matched MeSH terms: Pasteurella multocida/immunology*
Fulltext Complete Genome Sequence Analysis and Characterization of Selected Iron Regulation Genes of Pasteurella Multocida Serotype A Strain PMTB2.1

Jabeen S, Yap HY, Abdullah FFJ, Zakaria Z, Isa NM, Tan YC, et al.

Genes (Basel), 2019 01 25;10(2).
PMID: 30691021 DOI: 10.3390/genes10020081

Although more than 100 genome sequences of Pasteurella multocida are available, comprehensive and complete genome sequence analysis is limited. This study describes the analysis of complete genome sequence and pathogenomics of P. multocida strain PMTB2.1. The genome of PMTB2.1 has 2176 genes with more than 40 coding sequences associated with iron regulation and 140 virulence genes including the complete tad locus. The tad locus includes several previously uncharacterized genes such as flp2, rcpC and tadV genes. A transposable phage resembling to Mu phages was identified in P. multocida that has not been identified in any other serotype yet. The multi-locus sequence typing analysis assigned the PMTB2.1 genome sequence as type ST101, while the comparative genome analysis showed that PMTB2.1 is closely related to other P. multocida strains with the genomic distance of less than 0.13. The expression profiling of iron regulating-genes of PMTB2.1 was characterized under iron-limited environment. Results showed significant changes in the expression profiles of iron-regulating genes (p < 0.05) whereas the highest expression of fecE gene (281 fold) at 30 min suggests utilization of the outer-membrane proteins system in iron acquisition at an early stage of growth. This study showed the phylogenomic relatedness of P. multocida and improved annotation of important genes and functional characterization of iron-regulating genes of importance to the bacterial growth.

Matched MeSH terms: Pasteurella multocida/classification; Pasteurella multocida/genetics*; Pasteurella multocida/metabolism
Fulltext Draft Genome Sequence of Pasteurella multocida subsp. multocida Strain PMTB, Isolated from a Buffalo

Yap HY, Ghazali K, Wan Mohamad Nazarie WF, Mat Isa MN, Zakaria Z, Omar AR

Genome Announc, 2013;1(5).
PMID: 24136854 DOI: 10.1128/genomeA.00872-13

Pasteurella multocida serotypes B:2 and E:2 are the main causative agents of ruminant hemorrhagic septicemia in Asia and Africa, respectively. Pasteurella multocida strain PMTB was isolated from a buffalo with hemorrhagic septicemia and has been determined to be serotype B:2. Here we report the draft genome sequence of strain PMTB.

Matched MeSH terms: Pasteurella multocida
Fulltext First High-Quality Draft Genome Sequence of Pasteurella multocida Sequence Type 128 Isolated from Infected Bone

Kavousi N, Eng WW, Lee YP, Tan LH, Thuraisingham R, Yule CM, et al.

Genome Announc, 2016;4(2).
PMID: 26941132 DOI: 10.1128/genomeA.00023-16

We report here the first high-quality draft genome sequence of Pasteurella multocida sequence type 128, which was isolated from the infected finger bone of an adult female who was bitten by a domestic dog. The draft genome will be a valuable addition to the scarce genomic resources available for P. multocida.

Matched MeSH terms: Pasteurella multocida
Fulltext Complete Genome Sequence of Pasteurella multocida Serotype A Strain PMTB2.1 Isolated from Buffaloes That Died of Septicemia in Malaysia

Jabeen S, Yong YH, Abdullah FJF, Zakaria Z, Mat Isa N, Tan YC, et al.

Genome Announc, 2017 Nov 02;5(44).
PMID: 29097462 DOI: 10.1128/genomeA.01190-17

Pasteurella multocida causes pneumonic pasteurellosis and hemorrhagic septicemia (HS) in large ruminants. In this study, we determined the complete genome sequence of P. multocida strain PMTB2.1 capsular serotype A isolated from buffaloes that died of septicemia.

Matched MeSH terms: Pasteurella multocida
Fulltext Atypical contact lens related corneal ulcer caused by pasteurella multocida

Fatin Hanisah, F., Umi Kalthum, M. N., Rona Asnida, N., Jemaima, C. H.

Journal of Surgical Academia, 2018;8(1):43-46.
MyJurnal

A 55-year-old healthy lady with history of regular contact lens (CL) use presented with 10 days history of
progressive left eye blurring of vision, redness and pain. There was good CL hygiene practiced with no history of
swimming, trauma or contact with domestic pets. Left eye vision was hand movement and right eye was 1/60,
pinhole 6/18. On the left eye, there was a central, oval-shaped corneal infiltrate with an overlying large epithelial
defect and stromal oedema, with significant anterior chamber cells and fibrin. B-mode ultrasound showed no vitritis.
Intensive topical benzylpenicillin 10000iu/ml and topical gentamycin 1.4% hourly, homatropine 2% three times
daily, oral doxycycline and oral ascorbic acid were started. The gram stain results showed gram positive cocci
growth. Her ulcer improved with the treatment and preservative-free dexamethasone 0.1% once daily was
commenced to reduce inflammation and scarring. Interestingly, culture was reported as Pasteurella maltocida, a
gram negative bacilli sensitive to penicillin, and so treatment was continued until the ulcer completely healed. She
had central corneal scarring with best corrected vision of 6/24 in the left eye but was not keen on further surgery to
improve her vision. Although it has not been previously reported, Pasteurella multocida can cause CL related
corneal ulcer with severe anterior chamber inflammation. This diagnosis should be considered even if there is trivial
contact or no history of exposure to domestic animals.

Matched MeSH terms: Pasteurella multocida
Pathological changes in the respiratory tract of goats infected by Pasteurella multocida B:2

Shafarin MS, Zamri-Saad M, Khairani BS, Saharee AA

J Comp Pathol, 2009 Feb-Apr;140(2-3):194-7.
PMID: 19110260 DOI: 10.1016/j.jcpa.2008.10.005

Clinical and pathological changes are described in groups of five goats pretreated with dexamethasone and then infected with a large dose of Pasteurella multocida B:2 (the cause of haemorrhagic septicaemia) by the intratracheal, subcutaneous or intranasal route (groups A, B and C, respectively). In group A, two goats died (on day 1 and 4 post-inoculation); in group B three died (days 2, 5 and 14); and in group C one died (day 20). The infecting organism was recovered from the four goats that died within < or =5 days. The major pulmonary lesions included acute pneumonia, congestion, oedema and hydrothorax. Subcutaneous oedema of the lower jaw and brisket, typically seen in cattle and buffalo, was absent in goats.

Matched MeSH terms: Pasteurella multocida
Experimental transmission of Pasteurella multocida 6:B in goats

Shafarin MS, Zamri-Saad M, Jamil SM, Siti Khairani B, Saharee AA

J Vet Med A Physiol Pathol Clin Med, 2007 Apr;54(3):136-9.
PMID: 17381677

Haemorrhagic septicaemia (HS) is an acute disease of cattle and buffaloes caused by Pasteurella multocida 6:B. Outbreaks of the disease have been closely associated with carrier animals that transmit the organism to susceptible animals during stressful condition. This study was conducted to determine whether goats exposed intranasally to P. multocida 6:B can transmit the organism to contact goats. Thirty-six healthy local Katjang goats were divided into four groups and goats of groups 1 and 3 were each inoculated intranasally with a 1-ml inoculum that contained 1 x 10(9) CFU/ml of live P. multocida 6:B. Following the exposure, all goats of groups 3 and 4 were injected with dexamethasone at the rate of 1 mg/kg for three consecutive days. At the end of the dexamethasone treatment, goats of groups 1 and 2 were commingled but kept separate from goats of groups 3 and 4, which were commingled in another pen. Three surviving goats from each group were killed on days 7, 14 and 21 post-exposure for postmortem examination. Naso-pharyngeal mucus and heart blood were collected on swabs. Tissues from lungs, lymph nodes and tonsils were collected for bacteriological isolation and identification. Only one goat of group 3 died 6 days post-exposure showing clinical signs and lesions typical of HS. Other goats showed mild signs of upper respiratory tract infection. Goats of all groups developed acute mild pneumonic lesions, however, those treated with dexamethasone had significantly (P < 0.05) more extensive lesion scoring based on the lesion scoring system. P. multocida 6:B was isolated from the nasal mucosa and lung lesions of exposed and contact goats not treated with dexamethasone. Exposed and contact goats treated with dexamethasone carried the organism for 21 days. P. multocida isolation from heart blood was made only from exposed and contact goats treated with dexamethasone. P. multocida was isolated from the lymph node of the goat that died during the experiment.

Matched MeSH terms: Pasteurella multocida/pathogenicity*
Fulltext In vitro treatment of lipopolysaccharide increases invasion of Pasteurella multocida serotype B:2 into bovine aortic endothelial cells

Yap SK, Zakaria Z, Othman SS, Omar AR

J Vet Sci, 2018 Mar 31;19(2):207-215.
PMID: 28693312 DOI: 10.4142/jvs.2018.19.2.207

Pasteurella multocida serotype B:2 causes hemorrhagic septicemia in cattle and buffalo. The invasion mechanism of the bacterium when invading the bloodstream is unclear. This study aimed to characterize the effects of immunomodulatory molecules, namely dexamethasone and lipopolysaccharide, on the invasion efficiency of P. multocida serotype B:2 toward bovine aortic endothelial cells (BAECs) and the involvement of actin microfilaments in the invasion mechanism. The results imply that treatment of BAECs with lipopolysaccharide at 100 ng/mL for 24 h significantly increases the intracellular bacteria number per cell (p ＜ 0.01) compared with those in untreated and dexamethasone-treated cells. The lipopolysaccharide-treated cells showed a significant decrease in F-actin expression and an increase in G-actin expression (p ＜ 0.001), indicating actin depolymerization of BAECs. However, no significant differences were detected in the invasion efficiency and actin filament reorganization between the dexamethasone-treated and untreated cells. Transmission electron microscopy showed that P. multocida B:2 resided in a vacuolar compartment of dexamethasone-treated and untreated cells, whereas the bacteria resided in cellular membrane of lipopolysaccharide-treated cells. The results suggest that lipopolysaccharide destabilizes the actin filaments of BAECs, which could facilitate the invasion of P. multocida B:2 into BAECs.

Matched MeSH terms: Pasteurella multocida/drug effects; Pasteurella multocida/pathogenicity*
Intranasal inoculation of recombinant DNA vaccine ABA392 against haemorrhagic septicaemia disease

Kang TL, Chelliah S, Velappan RD, Kabir N, Mohamad J, Nor Rashid N, et al.

Lett Appl Microbiol, 2019 Nov;69(5):366-372.
PMID: 31508837 DOI: 10.1111/lam.13215

We evaluate the efficacy of recombinant DNA vaccine ABA392 against haemorrhagic septicaemia infection through intranasal administration route by targeting the mucosal immunity. The DNA vaccine was constructed and subjected to animal study using the Sprague Dawley (SD) rat. The study was divided into two major parts: (i) active and (ii) passive immunization studies, involving 30 animals for each part. Each group was then divided into five test groups: two test samples G1 and G2 with 50 and 100 µg ml-1 purified DNA vaccine; one positive control G5 with 106 CFU per ml formalin-killed PMB2; and two negative controls, G3 and G4 with normal saline and pVAX1 vector. Both studies were conducted for the determination of immunogenicity by total white blood cell count (TWBC), indirect ELISA and histopathological changes for the presence of the bronchus-associated lymphoid tissue (BALT). Our findings demonstrate that TWBC, IgA and IgG increased after each of the three vaccination regimes: groups G1, G2 and G5. Test samples G1 and G2 showed significant differences (P

Matched MeSH terms: Pasteurella multocida/genetics; Pasteurella multocida/immunology*
Clinico-pathology, hematology and biochemistry responses in buffaloes towards Pasteurella multocida type B: 2 immunogen lypopolysaccharide via oral and intravenous routes of infection

Chung EL, Abdullah FF, Ibrahim HH, Marza AD, Zamri-Saad M, Haron AW, et al.

Microb Pathog, 2016 Feb;91:141-54.
PMID: 26706347 DOI: 10.1016/j.micpath.2015.12.003

Haemorrhagic septicaemia is a disease caused by Pasteurella multocida serotype B: 2 and E: 2. The organism causes acute, highly fatal septicaemic disease with high morbidity and mortality in cattle and more susceptible in buffaloes. Lipopolysaccharide can be found on the outer cell wall of the organism. Lipopolysaccharide is released during multiplication which leads to inflammatory reaction. It represents the endotoxin of P. multocida type B: 2 and responsible for toxicity in haemorrhagic septicaemia which plays an important role in the pathogenesis of the disease. Therefore, the aim of this study was to investigate the clinical signs, blood parameters, gross post mortem lesions and histopathology changes caused by P. multocida type B:2 immunogen lipopolysaccharide infections initiated through intravenous and oral routes of infection. 9 buffalo heifers were divided equally into 3 treatment groups. Group 1 was inoculated orally with 10 ml of phosphate buffer saline (PBS); Group 2 and 3 were inoculated with 10 ml of lipopolysaccharide broth intravenously and orally respectively. For the clinical signs, there were significant differences (p < 0.05) in temperature between the control, intravenous and oral group. In hematology and biochemistry findings, there were significant differences (p < 0.05) in erythrocytes, haemoglobin, PCV, MCV, lymphocytes, monocytes, eosinophils, GGT and albumin between the control, intravenous and oral group. However, there were no significant differences (p > 0.05) in the MCHC, leukocytes, band neutrophils, basophils, thrombocytes, plasma protein, icterus index, total protein, globulin and A:G ratio between intravenous and oral group. For Group 2 buffaloes, there were gross lesions in the lung, trachea, heart, liver, spleen, and kidney. In contrast, lesions were only observed in the lung, trachea and liver of Group 3 buffaloes. There were significant differences (p < 0.05) in hemorrhage and congestion; necrosis and degeneration; and inflammatory cells infiltration between experimental groups and control group. However, there were no significant differences (p > 0.05) in edema lesion between groups. In conclusion, this study is a proof that oral route infection of P. multocida type B:2 immunogen lipopolysaccharide can be used to stimulate host cell responses where oral vaccine through feed could be developed in the near future.

Matched MeSH terms: Pasteurella multocida/immunology*; Pasteurella multocida/pathogenicity; Pasteurella multocida/physiology
Comparative clinicopathological changes in buffalo and cattle following infection by Pasteurella multocida B:2

Annas S, Zamri-Saad M, Jesse FF, Zunita Z

Microb Pathog, 2015 Nov;88:94-102.
PMID: 26298001 DOI: 10.1016/j.micpath.2015.08.009

Haemorrhagic septicaemia (HS) is an acute, septicaemic disease of cattle and buffalo of Asia and Africa caused by Pasteurella multocida B:2 or E:2. Buffaloes are believed to be more susceptible than cattle. In this study, 9 buffaloes of 8 months old were divided equally into 3 groups (Groups 1, 3, 5). Similarly, 9 cattle of 8 months old were equally divided into 3 groups (Groups 2, 4, 6). Animals of Groups 1 and 2 were inoculated with PBS while Groups 3 and 4 were inoculated subcutaneously with 10(5) cfu/ml of P. multocida B:2. Animals of Groups 5 and 6 were inoculated intranasally with the same inoculum. Both buffaloes and cattle that were inoculated subcutaneously succumbed to the infection at 16 h and 18 h, respectively. Two buffaloes that were inoculated intranasally (Group 5) succumbed at 68 h while the remaining cattle and buffaloes survived the 72-h study period. Endotoxin was detected in the blood of infected cattle (Group 4) and buffaloes (Groups 3 and 5) prior to the detection of P. multocida B:2 in the blood. The endotoxin was detected in the blood of buffaloes of Group 3 and cattle of Group 4 at 0.5 h post-inoculation while buffaloes of Group 5 and cattle of Group 6 at 1.5 h. On the other hand, bacteraemia was detected at 2.5 h in buffaloes of Group 3 and cattle of Group 4 and at 12 h in buffaloes of Group 5 and cattle of Group 6. Affected cattle and buffaloes showed lesions typical of haemorrhagic septicaemia. These included congestion and haemorrhages in the organs of respiratory, gastrointestinal and urinary tracts with evidence of acute inflammatory reactions. The severity of gross and histopathology lesions in cattle and buffalo calves that succumbed to the infection showed insignificant (p > 0.05) difference. However, inoculated buffalo and cattle that survived the infection showed significantly (p < 0.05) less severe gross and histopathological changes than those that succumbed. In general, cattle are more resistant to intranasal infection by P. multocida B:2 than buffaloes.

Matched MeSH terms: Pasteurella multocida
Involvement of the nervous system following experimental infection with Pasteurella multocida B:2 in buffalo (Bubalus bubalis): A clinicopathological study

Marza AD, Jesse FF, Ahmed IM, Teik Chung EL, Ibrahim HH, Zamri-Saad M, et al.

Microb Pathog, 2016 Apr;93:111-9.
PMID: 26850845 DOI: 10.1016/j.micpath.2016.01.025

Haemorrhagic septicaemia (HS) is an acute, fatal, septicaemic disease of cattle and buffaloes caused by one of two specific serotypes of Pasteurella multocida B:2 and E:2 in Asian and African, respectively. It is well known that HS affect mainly the respiratory and digestive tracts. However, involvement of the nervous system in pathogenesis of HS has been reported in previous studies without details. In this study, nine buffalo calves of 8 months old were distributed into three groups. Animals of Group 1 and 2 were inoculated orally and subcutaneously with 10 ml of 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while animals of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. All calves in Group 1 and Group 3 were euthanised after 504 h (21 day) post-infection, while calves in Group 2 had to euthanise after 12 h post-infection as they develop sever clinical signs of HS. Significant differences were found in Group 2 in the mean scores of clinical signs, gross and histopathological changes which mainly affect different anatomic regions of the nervous system. In addition, successful bacterial isolation of P. multocida B:2 were obtained from different sites of the nervous system. On the other hand, less sever, clinical, gross and histopathological changes were found in Group 1. These results provide for the first time strong evidence of involving of the nervous system in pathogenesis of HS, especially in the peracute stage of the disease.

Matched MeSH terms: Pasteurella multocida
The ABA392/pET30a protein of Pasteurella multocida provoked mucosal immunity against HS disease in a rat model

Kang TL, Velappan RD, Kabir N, Mohamad J, Rashid NN, Ismail S

Microb Pathog, 2019 Mar;128:90-96.
PMID: 30584901 DOI: 10.1016/j.micpath.2018.12.042

Haemorrhagic septicaemia (HS) is a well-known high fatality septicaemic disease happening among bovines. The disease is caused by the Pasteurella multocida serotype B:2 bacteria. P. multocida B:2 has high mortality and morbidity rates and is spread through the intranasal and oral routes in bovines. In this study, our aim was to investigate the efficacy of the recombinant protein vaccine, ABA392/pET30a via intranasal inoculation by targeting the mucosal immunity. The constructed recombinant protein vaccine ABA392/pET30a was subjected to an animal study using Sprague Dawley rats. The study was divided into two parts: active and passive immunization studies. Both studies were carried out through the determination of immunogenicity (using Total White Blood Cell (TWBC) Count with Indirect ELISA) and histopathogenicity, analyzing (Bronchus Associated Lymphoid Tissue (BALT) formation) in lungs. As a result, the IgA and IgG development of both tested groups: group 1 (50μg/mL protein vaccine) and group 2 (100μg/mL protein vaccine) showed equivalent with the positive control group 4 (formalin-killed P. multocida B:2). However, there was a significant difference when compared with the negative control group 3 (normal saline). These results demonstrate that both the protein vaccine at the concentration 50μg/mL and 100μg/mL have the same efficacy as the commercially available positive control vaccine. From the studies, higher concentration of protein vaccine at 100μg/mL showed higher development of both IgA and IgG compared to 50μg/mL protein vaccine. Higher and rapid development of IgA compared to IgG showed that mucosal immunity has been induced through the intranasal administration of the protein vaccine. In addition, leucocytosis was observed at each dose of vaccination showed that the protein vaccine is capable to induce the immune responses of the host. Histopathogenicity studies of the vaccinated groups showed more BALT formation and no severe lesions after challenge compared to the negative control group. Besides, no inflammatory onsite or anaphylactic responses were observed after the intranasal inoculation which proved to be safer and provided longer lasting immunity. Therefore, recombinant protein vaccine ABA392/pET30a could be a potential candidate for intranasal administration which can provoke mucosal immunity against HS disease.

Matched MeSH terms: Pasteurella multocida
Iron-associated protein interaction networks reveal the key functional modules related to survival and virulence of Pasteurella multocida

Jatuponwiphat T, Chumnanpuen P, Othman S, E-Kobon T, Vongsangnak W

Microb Pathog, 2019 Feb;127:257-266.
PMID: 30550841 DOI: 10.1016/j.micpath.2018.12.013

Pasteurella multocida causes respiratory infectious diseases in a multitude of birds and mammals. A number of virulence-associated genes were reported across different strains of P. multocida, including those involved in the iron transport and metabolism. Comparative iron-associated genes of P. multocida among different animal hosts towards their interaction networks have not been fully revealed. Therefore, this study aimed to identify the iron-associated genes from core- and pan-genomes of fourteen P. multocida strains and to construct iron-associated protein interaction networks using genome-scale network analysis which might be associated with the virulence. Results showed that these fourteen strains had 1587 genes in the core-genome and 3400 genes constituting their pan-genome. Out of these, 2651 genes associated with iron transport and metabolism were selected to construct the protein interaction networks and 361 genes were incorporated into the iron-associated protein interaction network (iPIN) consisting of nine different iron-associated functional modules. After comparing with the virulence factor database (VFDB), 21 virulence-associated proteins were determined and 11 of these belonged to the heme biosynthesis module. From this study, the core heme biosynthesis module and the core outer membrane hemoglobin receptor HgbA were proposed as candidate targets to design novel antibiotics and vaccines for preventing pasteurellosis across the serotypes or animal hosts for enhanced precision agriculture to ensure sustainability in food security.

Matched MeSH terms: Pasteurella multocida/genetics*; Pasteurella multocida/physiology

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