Self-Assessment of Gram Staining Skill Using Video Recording in a Classroom Environment for Veterinary Nursing Students.

Practical skills are essential in the veterinary nursing curriculum. Given the increasing implementation of video recording in higher education, this study explored the feasibility and benefits of video recording as a classroom tool in professional education. Concerns regarding the inability to monitor individual students' performance during their laboratory course promoted the implementation of video recording-a blended learning method-in a veterinary nursing course. The approach was personalized for this study, particularly for the Gram staining skill. Students submitted video recordings demonstrating the progression of their skills development, and the instructor reviewed the recordings for assessment. The Participant Perception Indicator, a self-assessment, was used to determine students' experience, knowledge, and confidence gained after performing the skill. Video recording helped students to identify areas for self-improvement. It is also a helpful tool for instructors to ensure that students are meeting the learning standards. The results suggest that the use of video recording in learning Gram staining skills was effective. The evidence-based approach maximized students' learning and engagement, and it improved individualized assessment by the instructor and enabled the instructor to provide feedback on students' performance. During this period of increasing reliance on online teaching and learning, video recording in a classroom environment could be more widely used by instructors.

Leukotoxin production by Fusobacterium necrophorum strains in relation to severity of liver abscesses in cattle.

Fusobacterium necrophorum, a Gram-negative anaerobe, is the primary etiologic agent of liver abscesses of beef cattle. The bacterium, a member of the microbial community of the rumen, travels to the liver via portal circulation to cause abscesses. The severity of liver abscesses vary from mild with one or two small abscesses to severe with medium to large multiple abscesses. Leukotoxin, a secreted protein, is the critical virulence factor involved in the infection. Our objective was to compare leukotoxin production between strains of F. necrophorum isolated from mild and severe liver abscesses collected from slaughtered cattle. The quantification of leukotoxin was based on assays to measure cytotoxicity and protein antigen concentration. One-hundred strains, 50 from mild and 50 from severe abscesses, were utilized in the study. Cell-free supernatants were prepared from cultures grown in anaerobic broth at 9 and 24 h incubations. The leukotoxic activity was quantified by measuring cytotoxicity based on the release of lactic dehydrogenase from bovine lymphocyte cells, BL3, treated with the culture supernatant. Leukotoxin protein concentration was quantified by a sandwich ELISA assay with a leukotoxin-specific monoclonal antibody as the capture antibody. The leukotoxin activity and concentration were highly variable among the strains within each severity of liver abscesses. Although the leukotoxic activity was unaffected by incubation time, leukotoxin protein concentration was consistently higher at 24 h compared to 9 h incubation. Strains from severe liver abscesses had significantly higher leukotoxic activity and higher protein concentration compared to strains from mild liver abscesses (P < 0.0001) at both 9 and 24 h culture supernatants. Across all strains, the correlation coefficients between leukotoxic activity and leukotoxin concentration at 9 and 24 h were 0.14 (P = 0.17) and 0.47 (P < 0.0001), respectively. In conclusion, strains isolated from severe liver abscesses had significantly higher leukotoxic activities and leukotoxin protein concentrations compared to strains isolated from mild liver abscesses.

Leukotoxic activity of Fusobacterium necrophorum of cattle origin.

Fusobacterium necrophorum is a Gram negative, rod-shaped and aero tolerant anaerobe. In animals, it is an opportunistic pathogen frequently associated with necrotic infections, generally called necrobacillosis, such as calf diphtheria, foot rot and liver abscesses in cattle. Two subspecies exist: subsp. necrophorum and subsp. funduliforme. Among several virulence factors, leukotoxin (Lkt) is considered to be a major factor and a protective antigen. The objective of the study was to utilize BL3 cells and measure the release of lactic dehydrogenase to quantify Lkt activity of F. necrophorum. The assay was used to examine the effects of storage and handling conditions, growth media, polymyxin B addition on the cytotoxicity and evaluate Lkt activities of F. necrophorum strains isolated from bovine liver abscesses and foot rot. The Lkt activity peaked at 9 h of incubation. There was a significant decrease in the cytotoxicity measured in the samples after each freeze and thaw cycle. No difference was observed in the cytotoxicity for the samples handled aerobically versus anaerobically. Lkt activities of strains grown in anaerobic Brain-Heart Infusion broth were higher compared to Vegitone broth. A small reduction in the cytotoxicity activity was observed after the addition of polymyxin. The Lkt activity was consistently higher in strains of subsp. necrophorum than subsp. funduliforme of liver abscess origin. Among the strains isolated from cattle foot rot, Lkt activities of subsp. necrophorum strains appear to be much more variable. Use of BL3 cells in combination of lactic acid dehydrogenase assay appears to be a simple and valid assay to measure Lkt activity of F. necrophorum.

Role of the stress-associated chemicals norepinephrine, epinephrine and substance P in dispersal of Mannheimia haemolytica from biofilms.

Bovine respiratory disease (BRD) is a major problem for the cattle industry that is triggered by various environmental stressors, pathogens and host responses. Mannheimia hemolytica, an important bacterial component of BRD, are present within the nasopharayngeal region of normal calves as commensal biofilm communities. However, following stress there are changes in the nasopharyngeal microenvironment that triggers the transition of the commensal M. haemolytica into a pulmonary pathogen. The factors responsible for this transition in- vivo are unknown. In this study we developed an in-vitro biofilm model and investigated the effect of three stress- related compounds: norepinephrine (NE), epinephrine (E), and substance P (SP) on M. haemolytica biofilms. Biofilm formation was demonstrated for 3 bovine nasal isolates of M. haemolytica by growing them in basal culture media, basal media with additional glucose, and basal media with a reduced pH. Increased glucose enhanced biofilm biomass for 2/3 isolates, but acidic media did not increase biofilm biomass when compared to biofilm biomass in basal media. When the biofilm was exposed to NE, E and SP, there was a dispersal of the biofilm which was most effective with E, followed by NE, and SP being the least effective. Using high - throughput scanning electron microscopy and confocal-imaging we confirmed our experimental data that treatment with NE, E and SP cause dispersion of M.haemolytica from biofilms.