BACKGROUND: Annexins (including annexin A1 and annexin A2) are important proteins which have some roles in organisms such as intracellular signal conduction, membrane cellular skeletal connection, cellular proliferation and differentiation, especially inhibitory function in inflammation processing. Pasteurella multocida is the most common bacterial pathogen and has high prevalence in pneumonia. Objectives: This study was aimed to determine experimentally annexin A1 and annexin A2 in the serum of calves affected by Pasteurella multocida pneumonia. Methods: In this research, 10 male calves (2 - 4 months) were allocated to two equal groups, one group as the case: 300 ml in dilution 2×109 CFU Pasteurella multocida bacteria and the other as control group: 300 ml normal saline inoculated by special lavage catheter through oral to trachea. Clinical scores were recorded based on available tables. In treatment group, about 18 to 24 hours after inoculation and synchronous with observation clinical signs changes, bronchoalveolar lavage for cytology and bacteriology examination were done of fluids results from washing. Blood sampling was taken from calves jugular vein in both groups then blood serums were examined by using ELISA kits. Results: The rates of annexin A1 and annexin A2  in blood serum of treatment group showed significant increase (using independent t test) compared to control group (p<0.05). Conclusions: It seems these annexins (annexin A1 and annexin A2) can be used as important biomarkers in blood serum to diagnose inflammation processes such as pneumonia.

OBJECTIVE To develop 2 rapid loop-mediated isothermal amplification (LAMP) assays for detection of Pasteurella multocida DNA (Pm-LAMP assay) and P multocida DNA from strains associated with hemorrhagic septicemia (HS) in cattle and buffalo (HS-LAMP assay). SAMPLE Solutions containing 16 P multocida strains and 9 other bacterial species at various concentrations. PROCEDURES Optimal conditions were determined for running the Pm-LAMP and HS-LAMP assays. The assays were then used to detect DNA of the test organisms. Results of LAMP assays were validated against conventional PCR assays designed for specific detection of P multocida and the B:2 serotype of HS-associated strains. RESULTS Following incubation of sample reaction mixtures for 27 minutes, specificity and sensitivity of the HS-LAMP assay at template DNA amounts as low as 5 pg were 93% and 97%, respectively. When duplicates of each sample were incubated for 28 minutes (a positive result defined as positive results for both reactions of a given sample), specificity and sensitivity of the HS-LAMP assay in the same conditions increased to 100%. The best specificity and sensitivity of Pm-LAMP single (93% and 91%) and duplicate (97% and 98%) reactions at template DNA amounts as low as 10 pg were achieved at 33 and 34 minutes, respectively. CONCLUSIONS AND CLINICAL RELEVANCE These preliminary findings suggested the developed HS-LAMP assay had high sensitivity and specificity for detection of HS-associated P multocida. Additional research is needed to determine the accuracy of the assay for use on clinical specimens obtained in HS-endemic countries such as Pakistan and Thailand.

A total of 65 Pasteurella multocida were isolated and identified from various animal’s samples received by Veterinary Research Institute (VRI) during the period of 2014 to 2016. These animals comprises of cattle, goat, pig, chicken, duck and rabbit. The serogroup of Pasteurella multocida were carried out using designation system of Carter’s capsular typing and molecular serogrouping method. Based on cases submitted to VRI, the prevalence of pasteurellosis in Malaysia ranging from 1.0% to 3.2% (2014 to 2016). It is low compared to previous reports and the pattern of predominant serogroups and animal hosts were found to be changing every year. In 2014, 80% (12/15) of the isolates were Pasteurella multocida Carter’s type D where all were isolated from goats. In 2015, the predominant serogroup changed to Pasteurella multocida Carter’s type A with a prevalence rate of 40.6% (13/32) which were mostly isolated from duck and cattle. While for Pasteurella multocida Carter’s type D, the prevalence in 2015 reduced to 21.9% (7/32) compared to the previous year and it was isolated from various animal species. Interestingly, in 2015 there was one isolate of Pasteurella multocida Carter’s type B isolated from goat with no reported history of outbreak. In 2016, the prevalence of Pasteurella multocida Carter’s type A increased to 72.2% (13/18), with a high percentage (92.3%) infection in young calves showing clinical signs with high mortality and morbidity in infected farms. Furthermore, during these 3 years of study, 3 isolates of Pasteurella multocida serogroup F were also identified each from pig, goat and chicken, respectively. In conclusion, this study revealed that pasteurellosis had become sporadic in Malaysia and the distribution of serogroups were diverse in all species of animal with no definitive host.