Intranasal inoculation of recombinant DNA vaccine ABA392 against haemorrhagic septicaemia disease.

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 < 0·05) compared to the negative controls, G3 and G4, but no significant differences from the positive control G5. Groups G1, G2 and G5 showed more formation of BALT compared to the negative controls, G3 and G4. Our results show that intranasal inoculation of recombinant DNA vaccine ABA392 can provoke mucosal immunity which makes it a potential prophylactic against HS. SIGNIFICANCE AND IMPACT OF THE STUDY: New approach of combating haemorrhagic septicaemia disease among bovines by recombinant DNA vaccine is crucial to overcome the loss of edible products from the infected bovines. DNA vaccine can potentially serve as a better immunogen which would elicit both cellular and humoral immunity, and it is also stable for its molecular reproduction. This research report demonstrates an effective yet simple way of administering the DNA vaccine via the intranasal route in rats, to provoke the mucosal immunity through the development of immunoglobulins IgA, IgG and bronchus-associated lymphoid tissue which guard as the first-line defence at the host's mucosal lining.

A Recombinant Subunit HSABA392 as a potential Vaccine for Haemorrhagic septicaemia disease in livestock.

Haemorrhagic septicaemia (HS) is a major disease in cattle and buffaloes, caused by certain serotypes of Pasteurella multocida, mainly B and E serotypes. Frequent HS outbreak has a major impact in many Asian countries, including Malaysia, where farmers encounter economic loss due to low milk production as well as death of their livestock. There are four types of vaccines available; broth bacterins, alum precipitated vaccine, aluminium hydroxide gel vaccine and oil adjuvant vaccine (OAV), but these vaccines can only provide short term immunity and therefore need to be administered annually. Hence, the development of a protein vaccine using recombinant antigen can be a potential candidate for the production of HS vaccine that would give longer immunity. We have successfully cloned the ABA392 gene fragment into a protein expression vector, pET-30a. The protein was expressed from our ABA392/pET30a clone and the immunogenicity of the protein has been tested on rats. This vaccine was able to trigger an immune response and therefore has the potential to be tested as suitable vaccine candidate in future studies. It is envisaged that this subunit vaccine will make a significant contribution in the management of HS among livestock in future.

A Recombinant Subunit HSABA392 as a potential Vaccine for Haemorrhagic septicaemia disease in livestock

@#Haemorrhagic septicaemia (HS) is a major disease in cattle and buffaloes, caused by certain serotypes of Pasteurella multocida, mainly B and E serotypes. Frequent HS outbreak has a major impact in many Asian countries, including Malaysia, where farmers encounter economic loss due to low milk production as well as death of their livestock. There are four types of vaccines available; broth bacterins, alum precipitated vaccine, aluminium hydroxide gel vaccine and oil adjuvant vaccine (OAV), but these vaccines can only provide short term immunity and therefore need to be administered annually. Hence, the development of a protein vaccine using recombinant antigen can be a potential candidate for the production of HS vaccine that would give longer immunity. We have successfully cloned the ABA392 gene fragment into a protein expression vector, pET-30a. The protein was expressed from our ABA392/pET30a clone and the immunogenicity of the protein has been tested on rats. This vaccine was able to trigger an immune response and therefore has the potential to be tested as suitable vaccine candidate in future studies. It is envisaged that this subunit vaccine will make a significant contribution in the management of HS among livestock in future.

Radiation dose reduction in temporal bone CT with iterative reconstruction technique.

BACKGROUND AND PURPOSE: Iterative reconstruction of CT images is characterized by reduced image noise and may allow reduction in radiation exposure. We investigated the influence of an IRT technique on image quality and radiation dose savings when applied to temporal bone CT. MATERIALS AND METHODS: Based on the typical image quality level of adult subjects using routine radiation dose and FBP, an exsomatized cadaveric head with CNR characteristics closest to the level of clinical subjects was identified. Cadaver acquisitions were performed at multiple levels of tube current exposure. Reconstructions were performed using FBP and IRT (iDose), with multiple iDose levels applied for each acquisition. Transverse and coronal reformations of all reconstructions were evaluated subjectively and objectively. Phantom tests were performed to validate the protocol optimizations with iDose, specifically the spatial resolution relative to routine dose acquisitions. Finally, the results of protocol optimization with iDose were clinically validated in 50 patients. RESULTS: At the same radiation dose, the image CNR of iDose reconstructions was higher than that of FBP and progressively increased with higher iDose levels. The combination of 100 mAs/section and iDoseL5 was the lowest dose that met the requirements for diagnostic acceptability, with CNR slightly higher than our routine institution protocol of 200 mAs/section with FBP reconstruction. Spatial resolution characteristics were similar between FBP and iDose at all different strengths. The findings were consistent among the cadaver, phantom, and clinical acquisitions. CONCLUSIONS: The iDose IRT can help reduce radiation dose of temporal bone CT by 50% relative to routine institution protocols with FBP, while maintaining diagnostic image quality.

Assessment of neutrophil chemotaxis by laser and video densitometry.

A laser densitometer and a video densitometer were used to evaluate neutrophil chemotaxis slides which were also assessed by the standard microscopic technique. A linear relationship was observed between cell number per high power field (HPF) and peak height by laser densitometry (r2 = 0.66) or peak area by video densitometry (r2 = 0.81). For wells containing more than 20 cells/HPF the least variability was observed with video densitometry. Quantitative results from neutrophil chemotaxis assays performed in 48-well microchambers can be obtained rapidly and conveniently by the use of commercially available video densitometers.