Pasteurella Multocida Serotype Research Articles

Complete genome sequences of eight Pasteurella multocida isolates representing all lipopolysaccharide outer core loci.

Pasteurella multocida (PM) is a major bacterial pathogen that causes fowl cholera disease in both domestic poultry and wild birds. Here, we report the complete genome sequences of eight PM isolates representing all known lipopolysaccharide outer core loci, which are phenotypically expressed as 16 known PM serotypes.

Identification and Antibiotic Resistance of Pasteurella multocida Isolated from Infected Layer Chickens in West Java, Indonesia

Bacterial infections, such as those caused by Pasteurella multocida serotype A, pose significant threats to poultry farming. The use of antibiotics to treat these infections can lead to antibiotic resistance. The present study aimed to identify Pasteurella multocida from 14 Hisex Brown layer chicken hen farms, with chikens aged 25-55 weeks, in West Java, Indonesia, and to evaluate their resistance to various antibiotics. Three samples from each farm were collected from dead chickens having symptoms of fowl cholera. Initially, the study involved isolating and identifying isolates from liver, heart, and lung organs via polymerase chain reaction. The colony was then tested for antibiotic resistance using the disk diffusion method. The results showed that 13 samples were Pasteurella multocida and nine were serotype A. The test results also indicated that all isolates were resistant to colistin (10 µg) and sensitive to tetracycline (30 µg), amoxicillin (25 µg), enrofloxacin (5 µg), sulfamethoxazole (25 µg), lincomycin (109 µg), and ciprofloxacin (5 µg). The study concluded that none of the Pasteurella multocida type A isolates were any longer sensitive to colistin, with some isolates still sensitive to tetracycline, amoxicillin, enrofloxacin, sulfamethoxazole, lincomycin, and ciprofloxacin, and two isolates showing multidrug resistance patterns. Keywords: Antibiotic, Fowl cholera, Pasteurella multocida, Layer chicken

Pathogenicity and Genomic Characteristics Analysis of Pasteurella multocida Serotype A Isolated from Argali Hybrid Sheep.

Respiratory diseases arising from co-infections involving Pasteurella multocida (P. multocida) and Mycoplasma ovipneumoniae (Mo) pose a substantial threat to the sheep industry. This study focuses on the isolation and identification of the P. multocida strain extracted from the lung tissue of an argali hybrid sheep infected with Mo. Kunming mice were used as a model to assess the pathogenicity of P. multocida. Subsequently, whole genome sequencing (WGS) of P. multocida was conducted using the Illumina NovaSeq PE150 platform. The whole genome sequencing analysis involved the construction of an evolutionary tree to depict conserved genes and the generation of a genome circle diagram. P. multocida, identified as serotype A, was named P. multocida SHZ01. Our findings reveal that P. multocida SHZ01 infection induces pathological manifestations, including hemorrhage and edema, in mice. The phylogenetic tree of conserved genes analyzing P. multocida from different countries and different host sources indicates close relatedness between the P. multocida SHZ01 strain and the P. multocida 40540 strain (A:12), originating from turkeys in Denmark. The genome of P. multocida SHZ01 comprises 2,378,508 base pairs (bp) with a GC content of 40.89%. Notably, this strain, designated P. multocida, exhibits two distinct gene islands and harbors a total of 80 effector proteins associated with the Type III Secretion System (T3SS). The P. multocida SHZ01 strain harbors 82 virulence genes and 54 resistance genes. In the P. multocida SHZ01 strain, the proteins, genes, and related GO and KEGG pathways have been annotated. Exploring the relationship between these annotations and the pathogenicity of the P. multocida SHZ01 strain would be valuable. This study holds great significance in further understanding the pathogenesis and genetic characteristics of the sheep-derived P. multocida SHZ01 strain. Additionally, it contributes to our understanding of respiratory diseases in the context of co-infection.

A nose for trouble: ecotoxicological implications for climate change and disease in Saiga antelope (S. t. tatarica)

In recent decades, Saiga antelope (Saiga t. tatarica) mass die-offs have become more common. The mass die-off of 2015 in central Kazakhstan, recorded 140,000 individual deaths across multiple herds. Previously, research has shown atmospheric humidity, the bacterium Pasteurella multocida serotype B, and resultant haemorrhagic septicaemia, were the primary cause. However, other synergistic factors may have impacted this process. Here we use a multivariate compositional data analysis (CoDA) approach to assess what other factors may have been involved. We show a pollutant linkage mechanism where relative humidity and dewpoint temperature combine with environmental pollutants, potentially toxic elements (e.g., Hg, As), complex carbon compounds (e.g., Acetone, Toluene), and inorganic compounds (e.g., CHx, SO2) which affected the Saiga during the calving season (start and peak) and at the onset of the mass die-off. We suggest a mechanism for this process. Upon arrival at their carving grounds, the Saiga experienced a sudden precipitation event, a spike in temperatures, and resultant high humidity occurs. The infectious bacterium P. multocida serotype B then spreads. Further, environmental pollutants contained within steppe soils are released to the air, forming localised smog events, these synergistically combine, and mass die-off occurs.

Back passage of haemorrhagic septicaemia (HS) vaccine seed C82 in cattle calf

Haemorrhagic septicaemia (HS) is caused by Pasteurella multocida serotypes B: 2, a Gram-negative coccobacilli bacterium. It causes fatal septicaemia with high mortality in cattle and buffaloes. Disease control and prevention are by effective HS vaccination in animals. A study was conducted on one mixed breed cattle calf for back passage of haemorrhagic septicaemia (HS) seed strain C82. Prior to bacterial inoculation in the calf, Mouse Protection Test (MPT) was conducted to ensure the calf is free from any HS antibody. The calf was injected subcutaneously with 0.5 ml dilutions of 10-3 of the bacteria incubated overnight (approximately 106 CFU/ml). After ±30 hours inoculation, post-mortem was conducted on the calf showing sign of HS. Heart blood and organ samples were collected and cultured on blood agar to determine bacteria purity. Blood and organ samples were also sent to the bacteriology laboratory in VRI for confirmatory tests. The death of the calf is confirmed by re-isolation of Pasteurella multocida Type B. Heart blood collected was processed, freeze-dried and keep as vaccine seed for further usage in vaccine production. Revival of vaccine seed is needed to ensure its pathogenicity and functionality for production purposes.

Secretome Analysis of High- and Low-Virulent Bovine Pasteurella multocida Cultured in Different Media

Bovine Pasteurella multocida (P. multocida) serotype A is one of the major causes of bovine respiratory disease (BRD). We used data-independent acquisition (DIA) LC-MS/MS combined with bioinformatics analysis to identify proteins secreted by P. multocida. A total of 154 proteins were obtained from the supernatants of two isolates of bovine P. multocida serotype A (high virulent PmCQ2 and low virulent PmCQ6) cultured in Martin or BHI media, of which 50 were identified as putative secreted proteins. Further studies showed that Tuf, an elongation factor Tu, was highly expressed in P. multocida and secreted into infected tissues. Tuf stimulated strong innate immune responses of macrophages and had protective efficacy against P. multocida infection in a mouse model. The results provide insight into the secreted proteins of P. multocida and suggest new targets for vaccine development against P. multocida.

Pathology, virulence associated gene profiling, antimicrobial susceptibility and pathogenicity of untypeable capsular serotypes of Pasteurella multocida isolated from slaughtered pigs of India.

Pasteurella multocida is widely distributed in all pig-rearing countries, affecting the economic viability and profitability of pig production. The present research highlights the molecular characterization and pathology of untypeable capsular serotypes of P. multocida in slaughtered pigs from prominent pig-rearing states of India. The prevalence of Pasteurellosis was 27.17% by PM-PCR assay, while isolation rate was 7.62%. The microscopic lesions of bronchopneumonia, tonsillitis, and the presence of bacterial antigens in IHC confirmed P. multocida with pathologies. In capsular typing, the majority of the isolates were untypeable with prevalence of 52.15% and 43.58% in molecular and microbiological methods respectively. All the isolates showed the uniform distribution of virulence genes such as exbB, nanB, sodC, plpB, and oma87 (100%), while the variations were observed in ptfA, hasR, ptfA, pfhA, hsf-1, and plpE genes. The untypeable isolates showed higher prevalence of hsf-1 gene as compared to others. The untypeable serotypes showed a higher degree of resistance to ampicillin, amoxicillin, and penicillin antibiotics. The mouse pathogenicity testing of untypeable capsular isolates confirmed its pathogenic potential. The higher frequency of pathogenic untypeable isolates with antibiotic resistance profile might pose a serious threat to the pigs, and therefore, preventive measures should be adopted for effective control.

Genomic characterization and comparative genomic analysis of HS-associated Pasteurella multocida serotype B:2 strains from Pakistan

BackgroundHaemorrhagic septicaemia (HS) is a highly fatal and predominant disease in livestock, particularly cattle and buffalo in the tropical regions of the world. Pasteurella multocida (P. multocida), serotypes B:2 and E:2, are reported to be the main causes of HS wherein serotype B:2 is more common in Asian countries including Pakistan and costs heavy financial losses every year. As yet, very little molecular and genomic information related to the HS-associated serotypes of P. multocida isolated from Pakistan is available. Therefore, this study aimed to explore the characteristics of novel bovine isolates of P. multocida serotype B:2 at the genomic level and perform comparative genomic analysis of various P. multocida strains from Pakistan to better understand the genetic basis of pathogenesis and virulence.ResultsTo understand the genomic variability and pathogenomics, we characterized three HS-associated P. multocida serotype B:2 strains isolated from the Faisalabad (PM1), Peshawar (PM2) and Okara (PM3) districts of Punjab, Pakistan. Together with the other nine publicly available Pakistani-origin P. multocida strains and a reference strain Pm70, a comparative genomic analysis was performed. The sequenced strains were characterized as serotype B and belong to ST-122. The strains contain no plasmids; however, each strain contains at least two complete prophages. The pan-genome analysis revealed a higher number of core genes indicating a close resemblance to the studied genomes and very few genes (1%) of the core genome serve as a part of virulence, disease, and defense mechanisms. We further identified that studied P. multocida B:2 strains harbor common antibiotic resistance genes, specifically PBP3 and EF-Tu. Remarkably, the distribution of virulence factors revealed that OmpH and plpE were not present in any P. multocida B:2 strains while the presence of these antigens was reported uniformly in all serotypes of P. multocida.ConclusionThis study's findings indicate the absence of OmpH and PlpE in the analyzed P. multocida B:2 strains, which are known surface antigens and provide protective immunity against P. multocida infection. The availability of additional genomic data on P. multocida B:2 strains from Pakistan will facilitate the development of localized therapeutic agents and rapid diagnostic tools specifically targeting HS-associated P. multocida B:2 strains.

Identification of serotypes of Mannheimia haemolytica and Pasteurella multocida from pneumonic cases of sheep and goats and their antimicrobial sensitivity profiles in Borana and Arsi zones, Ethiopia

Respiratory diseases caused by Mannheimia haemolytica (M. haemolytica) and Pasteurella multocida (P. multocida) have been known to result in a considerable loss due to mortality and reduced production. This study aimed at isolation and identification of M. haemolytica and P. multocida associated with pneumonic pasteurellosis in sheep and goats using bacteriological and molecular techniques. Identification of serotypes of M. haemolytica and P. multocida was done using indirect haemagglutination test. The in vitro antimicrobial sensitivity profiles of the M. haemolytica were tested using standard disk diffusion method. A total of 52 and 78 nasal swabs were collected from pneumonic cases for bacterial isolation and identification in Borana and Arsi zone, respectively. Four hundred sera samples were collected for identification of serotypes. The results showed that 17 of 52 (32.69%; 95% CI 20.33, 47.11) nasal swabs collected from pneumonic animals in Borana yielded positive results for Pasteurella/Mannheimia species, 13 (25.00%; 95% CI 14.03, 38.95) of which were M. haemolytica. None of the samples yielded P. multocida. Twenty-three of 78 (29.49%; 95% CI 19.69, 40.89) nasal swabs collected at Arsi from pneumonic animals yielded positive results for M. haemolytica (17) and P. multocida (6). Secondary biochemical characterization revealed that 14 of the 17 isolates conform to M. haemolytica whereas none of the 6 isolates suspected to be P. mutocida were confirmed. Eleven (84.62%) isolates from Borana and 4 (28.57%) from Arsi were confirmed to be M. haemolytica using PCR targeting the Rpt2 genes. Assay for M. haemolytica serotype A1 revealed all belong to A1. None of the isolates with cultural and morphological features of P. multocida gave positive results by molecular assay. Serological assay identified three serotypes of M. haemolytica namely A1, A2 and A7 almost in all of the samples whereas P. multocida serotype A was detected in 78.75% of the samples. The M. haemolytica isolates tested for susceptibility to antibiotics showed resistance against Bacitracin (83.33%) and Penicillin (50.00%) while they were found susceptible to Gentamycin (100%), Chloramphenicol (100%) and Sulfamethoxazole (100%) and Tetracycline (83.33%). In conclusion, the results of the present study revealed the association of M. haemolytica with pneumonic pasteurellosis in sheep and goats and can be of use in vaccine development in Ethiopia. Nevertheless, further investigations and continuous monitoring of antimicrobial resistance and appropriate selection and prudent use of antimicrobials in livestock sector are required.

Genome wide CRISPR screen for Pasteurella multocida toxin (PMT) binding proteins reveals LDL Receptor Related Protein 1 (LRP1) as crucial cellular receptor.

PMT is a protein toxin produced by Pasteurella multocida serotypes A and D. As causative agent of atrophic rhinitis in swine, it leads to rapid degradation of the nasal turbinate bone. The toxin acts as a deamidase to modify a crucial glutamine in heterotrimeric G proteins, which results in constitutive activation of the G proteins and permanent stimulation of numerous downstream signaling pathways. Using a lentiviral based genome wide CRISPR knockout screen in combination with a lethal toxin chimera, consisting of full length inactive PMT and the catalytic domain of diphtheria toxin, we identified the LRP1 gene encoding the Low-Density Lipoprotein Receptor-related protein 1 as a critical host factor for PMT function. Loss of LRP1 reduced PMT binding and abolished the cellular response and deamidation of heterotrimeric G proteins, confirming LRP1 to be crucial for PMT uptake. Expression of LRP1 or cluster 4 of LRP1 restored intoxication of the knockout cells. In summary our data demonstrate LRP1 as crucial host entry factor for PMT intoxication by acting as its primary cell surface receptor.

Prediction of Pasteurella multocida serotypes based on whole genomic sequences

The serotypes of Pasteurella multocida were predicted based on whole genomic sequences (WGS) with specific genes of the capsular and liposaccharide (LPS) outer core polysaccharide regions as targets. A total of 56 strains were whole genomic sequenced and in addition all assembled genomes from NCBI were included for comparison. BIGSdb (Bacterial Isolate Genome Sequence Database) was installed on a Linux server and targets for capsular types A, B, D, E and F were defined as gene sequences of hyaD, bcbD, dcbF, ecbJ and fcbD, respectively and targets for LPS groups 1, 2, 3, 4, 5, 6, 7 and 8 were defined as gene sequences of pcgB, nctA, gatF, latB, rmlA, nctB, ppgB and natG, respectively. The serotypes of P. multocida were predicted from WGS by designating the capsular type and LPS group as well as subtype alleles to isolates. Comparisons between WGS predictions of capsular types and classical phenotypic typing showed correspondence in 87 % of cases whereas comparisons of WGS predictions of LPS groups to phenotypic typing corresponded for 82 % of the strains. In total 93 % and 94 % of the strains available with WGS could be capsular and LPS group typed, respectively. The server is free to access from https://ivsmlst.sund.ku.dk.

Pathological study of Pasteurella Multocida Recombinant Clone ABA392

Objective: In current study Molecular analysis of the recombinant clone ABA392 was carried out Methodology. In this study, detailed molecular analysis of the recombinant clone ABA392 were carried out by examining plasmid stability, plasmid profiling, determination of the plasmid molecular weight, and restriction endonuclease analysis. Results: From the experiments above, the plasmid size of ABA392 was estimated as 3.5 kb, while the size of the insert gene of ABA392 was estimated as 0.8 kb (double digested with PstI and SmaI restriction enzymes). A Pathogenicity study of recombinant clone ABA392 was carried out via a mouse virulence test and histopathological analysis, where it was found that mice, which were challenged with ABA392 and the parent strain of Pasteurella multocida serotype B strain PMB202, were lethal within 24 to 72 hours upon inoculation. Based on microscopic examination, the virulence effect was observed by marked haemorrhage in lung, liver, spleen, and kidney tissues. Conclusion: This study has also proven that the plasmid of recombinant clone ABA392 was stably maintained in the Escherichia coli JM101 host system as reported previously. A preliminary protein analysis via SDS-PAGE was performed and a distinct protein band was most likely to be detected, which was approximately 16kD. Nonetheless, further characterization of the recombinant clone ABA392 proteins needs to be done to determine the nature of these proteins, which are involved in the virulence mechanism of hemorrhagic septicemia. Keywords: Plasmid profiling; Pathogenicity; recombinant clone; Pasteurella multocida

Polymerase chain reaction to confirm biochemically characterization method of Pasteurella multocida isolate from fatal cases of Septicaemia epizootica in Nusa Tenggara Timur

Septicemia epizootica (SE) is a common fatal systemic disease in cattle and buffalo due to Pasteurella multocida serotype B:2 in South and Southeast Asia countries, including Indonesia. This infectious agent is generally considered an opportunistic pathogen and located in the nasopharynx of the animal. To support the disease’s diagnosis, an acceptable identification procedure must be established. This study was to confirm bovine P. multocida isolate that has been identified through biochemical approaches at the first step, with the Polymerase Chain Reaction method. The isolate was obtained from the fatal outbreak of cattle in Kupang in 2016 and subjected to be identified using biochemical characterization, but it was time consumed. The API 20NE was applied to identify the isolate and help to save time. The PCR result showed positive for 16 sRNA and kmt genes, both were classified as specific genes, and the capsular serotype was detected in less than 24 hours. It indicates that PCR confirms biochemical technique, and it is an appropriate and faster method in detecting pathogen agents than biochemical one.

Phenotypic and Molecular Characterization of the Capsular Serotypes of Pasteurella multocida Isolated from Pneumonic Cases of Cattle in Ethiopia

Background: Pasteurella multocida is a heterogeneous species and opportunistic pathogen associated with pneumonia in cattle. Losses due to pneumonia and associated expenses are estimated to be higher in Ethiopia with limited information about the distribution of capsular serotypes. Hence, this study was designed to determine the biotypes and capsular serotypes of P. multocida from pneumonic cases of cattle.Methods: A cross-sectional study with a purposive sampling method was employed in 400 cattle from April 2018 to January 2019. Nasopharyngeal swabs and lung tissue samples were collected from clinically suspected pneumonic cases of calves (n = 170) and adult cattle (n = 230). Samples were analyzed using the bacteriological and molecular assay.Result: Bacteriological analysis revealed isolation of 61 (15.25%) P. multocida subspecies multocida. Molecular assay targeting KMT1 gene (~460 bp) confirmed P. multocida species. Capsular typing revealed the presence of serogroup A (hyaD-hyaC) gene (~1044 bp) and serogroup D (dcbF) gene (~657 bp) in 56 (91.80%) and 5 (8.20%) isolates, respectively. Isolation of P. multocida A:3 highlights its prevalence in the study areas. Therefore, the current finding suggests further comprehensive studies on P. multocida capsular types and microbial diversity of respiratory infection in cattle to design an effective control strategy.

Transcriptomic Analysis of High- and Low-Virulence Bovine Pasteurella multocida in vitro and in vivo.

Pasteurella multocida is a gram-negative opportunistic pathogen that causes various diseases in poultry, livestock, and humans, resulting in huge economic losses. Pasteurella multocida serotype A CQ6 (PmCQ6) is a naturally occurring attenuated strain, while P. multocida serotype A strain CQ2 (PmCQ2) is a highly virulent strain isolated from calves. Compared with PmCQ2, it was found that bacterial loads and tissue lesions of lung tissue significantly decreased and survival rates significantly improved in mice infected with PmCQ6 by intranasal infection. However, comparative genome analysis showed that the similarity between the two strains is more than 99%. To further explore the virulence difference mechanism of PmCQ2 and PmCQ6, transcriptome sequencing analysis of the two strains was performed. The RNA sequencing analysis of PmCQ2 and PmCQ6 showed a large number of virulence-related differentially expressed genes (DEGs) in vivo and in vitro. Among them, 38 virulence-related DGEs were significantly up-regulated due to PmCQ6 infection, while the number of PmCQ2 infection was 46, much more than PmCQ6. In addition, 18 virulence-related DEGs (capsule, iron utilization, lipopolysaccharide, and outer membrane protein-related genes) were up-regulated in PmCQ2 infection compared to PmCQ6 infection, exhibiting a higher intensive expression level in vivo. Our findings indicate that these virulence-related DEGs (especially capsule) might be responsible for the virulence of PmCQ2 and PmCQ6, providing prospective candidates for further studies on pathogenesis.

PREPARATION AND COMPARATIVE EVALUATION OF HAEMORRHAGIC SEPTICAEMIA VACCINES USING EOLANE-150 AND EOLANE-170 AS OIL ADJUVANTS FOR CATTLE AND BUFFALO

Haemorrhagic septicaemia (HS) caused by Pasteurella multocida serotype B:2 is an economically important disease of cattle and buffalo, causes heavy economic losses due to sudden death of animals in developing countries like Pakistan. In this country, animals are vaccinated by alum (adjuvant) precipitated vaccine twice a year. Immunity induced through this prophylactic measure lasts for 3-4 months only. Two new HS oil based vaccines were prepared by using two new oil adjuvants such as Eolane-150 and Eolane-170. The ratio of bacterial antigen and oil adjuvants was 1:1 while bacterial dry weight was adjusted to 2 mg/ml. The addition of enrichments and aeration resulted in dense bacterial growth of Pasteurella multocida. Both new vaccines passed sterility, safety and potency tests as per OIE, 2017. Active and passive Mouse Protection Tests were performed to evaluate its potency. Indirect Haem-Agglutination (IHA) test was conducted on serum samples of two rabbits, groups each of which was vaccinated with HS oil based vaccines adjuvanted with Eolane-150 and Eolane-170. IHA indicated that immune response was higher (GMT=32) initially on 45th day to 75th day post vaccination and then declined (GMT=16) in the rabbits vaccinated with HS vaccine adjuvanted with Eolane-150, while protective immune response remained constant (GMT=16) up to ninety days post vaccination in the rabbits vaccinated with HS vaccine adjuvanted with Eolane-170. Vaccines were easy to inject with no side effects, including swelling at the injection site and longer protection as well. That would hopefully motivate the livestock owners and farmers to use this new product to protect their animals against this fatal HS disease.

Clinicopathological and Hematological Changes in Consequence to Experimental Infection of Rabbits with Pasteurella Multocida Type A

The current research aimed to study clinical, hematological and pathological changes occurred due to experimentally challenged rabbits with Pasteurella multocida type A. Twelve healthy Pasteurella multocida free rabbits were divided into control and challenged groups. Rabbits of the challenged group were intra-nasally inoculated with 0.5 ml of 1x105 CFU of Pasteurella multocida type A, while rabbits of the control group were inoculated with 0.5 ml of brain heart infusion broth (BHI). Clinical signs, hematological parameters and histopathological lesions were recoded for 14 days. Rabbits of challenged group showed nasal rubbing with minimal mucus discharge, conjunctivitis and reduction in spontaneous activity, but no mortality occurred. Hematologically, there was highly significant low RBCs count with concomitant significantly reduction of PCV in challenged rabbits, but hemoglobin concentration was insignificantly low in challenged animals as compared to control. Although, there was insignificant increase of WBCs count in challenged group, neutrophils and monocytes recorded significant and highly significant high value in challenged rabbits than control, respectively. Challenged rabbits, however demonstrated highly significant lymphopenia, but other blood parameters did not show any significant differences between challenged and control rabbits. Histopathologically, challenged animals showed meningitis, congestion and degenerative changes in respiratory epithelium, bronchopneumonia, vacuolar degeneration of hepatocytes and vascular changes in heart and spleen. Conclusively, the current study verified that challenging rabbits with Pasteurella multocida serotype A was not lethal and although it was essentially pneumotropic in nature, it caused lesions in other visceral organs, which in absence of bacteremia, could be caused by bacterial endotoxin liberated during disease pathogenesis.

Comparision of iron-regulated outer membrane proteins (IrOMP) and iron-sufficient outer membrane proteins (IsOMP) of Pasteurella multocida strains of porcine origin

An investigation was carried out to compare the effect of growth of P. multocida type A of pig origin in iron restricted and in iron sufficient media on the basis of their outer membrane protein extract. Pasteurella multocida serotype A was cultured in two ways. In one process only BHI medium was used and on the other process same medium was used which was supplemented with 2, 2’- Dipyridyl (iron-restricted medium). Both these cultures were used to obtain Outer Membrane Proteins (OMPs) by extraction of bacterial cells with 1% Sarcosyl. Separation of the OMPs in SDS-PAGE showed that OMPs were mixture of protein fraction with molecular weight ranging from 110 to 22.6 kDa in case of iron-restricted OMP and 47.3 to 29.9 kDa in case of iron sufficient OMP. The OMP with molecular masses 29.9 kDa grew in both iron-restricted as well as in iron sufficient medium.

Pathological findings in a fatal pet rabbit Pasteurellosis

A case of fatal Pasteurellosis infection in a 2-month-old rabbit is described. Pasteurellosis is considered one of the most important diseases of rabbits and has been reported as a constant serious and highly contagious disease of domestic rabbits. Before death, the main clinical signs observed were rhinitis with nasal discharge, abdominal breathing and coughing. At necropsy, pathological findings revealed severe pleuropneumonia with the accumulation of a remarkable amount of fibrinopurulent exudate in the thoracic cavity, serous rhinitis and tracheitis. A bacterial isolate from lungs was identified as Pasteurella multocida serotype A. The present study describes the histopathological lesions associated with P. multocida infection in a pet rabbit brought to visit with respiratory signs.

Molecular characterization of Pasteurella multocida pfhaB1 gene fragment from buffalo and cattle isolates from Nusa Tenggara Timur Indonesia

Almost all regions in Nusa Tenggara Timur East Nusa Tenggara (NTT) Province Indonesia are endemic areas of Haemorragic Septicaemia (HS), which is caused by Pasteurella multocida (P. multocida ) Serotypes B: 2. The fragment pfhaB1gene is one of the virulence factors of P. multocida.The objective of this study was to determaine the phylogenetic, homology of P. multocidapfhaB1gene fragment of isolatedfrom Buffalo and Cattle in NTT. The P. multocida isolateswere re-cultured and further microscopic examined the biochemical tests, PCR, sequencing, homology, and phylogenetic relatedness test. P. multocida was observed as gram negative, coccobacilus, no growth on MacConkey Agar, does not produce H2S and gas, nonmotile and indole positive, does not produce urease enzymes, does not use citrate as a carbon source, does not ferment maltose and lactose but it does ferment glucose, sucrose and mannitol. ThepfhaB1gene fragmentfrom buffalo and cattle NTT isolates and also Katha strain vaccine showed DNA band 506 bp. P.multocida isolates from buffalo and cattle in NTT have 91% - 99% score homology with the comparative isolate. The isolate P. multocida from buffalo and cattle in NTT are in one cluster and their phylogenetic relatedness is close to isolates from Iran and India. It is concluded that the pfhaB1gene fragmentof P. multocida from buffalo and cattle isolates have phylogenetic relatedness close and homolog with the other comparative isolates. Keywords: Haemorrhagic Septicaemia; Nusa Tenggara Timur; Pasteurella multocida; pfhaB1gene