Evaluation of the Bachelor of Veterinary Medicine (BVM) Curriculum at Sokoine University of Agriculture in Tanzania: Mapping to OIE Veterinary Graduate 'Day 1 Competencies'.

The World Organisation for Animal Health (OIE) provides the requirements needed for graduating veterinary professionals to be competent in the delivery of animal health services. However, significant differences in veterinary curricula across countries-attributable to differing animal health priorities and predominant types of veterinary practice-provide a challenge for veterinary schools to address these competencies adequately. As part of the OIE's veterinary education establishment Twinning Project activities, the College of Veterinary Medicine and Biomedical Sciences (CVMBS) of Sokoine University of Agriculture (SUA) in Tanzania undertook a curriculum mapping and gap analysis to assess the extent to which the veterinary curriculum addresses OIE's 'Day 1 Competencies' for graduating veterinarians. Results of the analysis indicated that all the OIE's Day 1 Competencies (general, specific, and advanced) are addressed to some degree by the courses present in the curriculum. However, gaps in the depth and breadth of instruction were found for a number of competencies in all three categories. These findings indicate a need for addressing the gaps in the next curriculum review. This will allow the development of a stronger curriculum that will efficiently meet the national and international animal health requirements.

Characterization of Mannheimia haemolytica in beef calves via nasopharyngeal culture and pulsed-field gel electrophoresis.

Mannheimia haemolytica is a major bacterial component of bovine respiratory disease (BRD); unfortunately, very little is known about M. haemolytica transmission dynamics among cattle. Identifying potential variation in M. haemolytica populations over time and induction of nasopharyngeal colonization and subsequent shedding are 2 areas where knowledge is lacking. In our study, 2 separate loads of 20 mixed-origin, male calves were purchased through an order buyer on different dates. Deep nasopharyngeal cultures (NPC) were performed on all calves on arrival and, if M. haemolytica-negative, a second screening culture was obtained. Calves that were negative on 2 initial NPCs (NEG; n = 4) were subsequently challenged with a previously isolated field strain of M. haemolytica in both the upper and lower respiratory tract, individually housed, and then monitored for M. haemolytica shedding via NPCs at 0.5, 1, 3, 5, 7, and 9 days postchallenge. Naturally M. haemolytica-positive calves (2 per load) were kept for additional daily cultures (POS; n = 4). Individual calf M. haemolytica status for both the POS and NEG groups was inconsistent between study days. Additionally, pulsed-field gel electrophoresis performed on isolates from the positive cultures showed that the NEG calves did not shed the M. haemolytica challenge strain, but rather 2 distinct clusters of M. haemolytica were shared among POS and NEG calves regardless of their initial status. Although sample sizes were small, these findings illustrate how variable the results of a single nasopharyngeal swab can be and the challenges of using an individual culture to truly represent animal M. haemolytica status.

Comparison of anamnestic responses to rabies vaccination in dogs and cats with current and out-of-date vaccination status.

OBJECTIVE: To compare anamnestic antibody responses of dogs and cats with current versus out-of-date vaccination status. DESIGN: Cross-sectional study. ANIMALS: 74 dogs and 33 cats. PROCEDURES: Serum samples were obtained from dogs and cats that had been exposed to rabies and brought to a veterinarian for proactive serologic monitoring or that had been brought to a veterinarian for booster rabies vaccination. Blood samples were collected on the day of initial evaluation (day 0) and then again 5 to 15 days later. On day 0, a rabies vaccine was administered according to label recommendations. Paired serum samples were analyzed for antirabies antibodies by means of a rapid fluorescent focus inhibition test. RESULTS: All animals had an antirabies antibody titer ≥ 0.5 IU/mL 5 to 15 days after booster vaccination. Dogs with an out-of-date vaccination status had a higher median increase in titer, higher median fold increase in titer, and higher median titer following booster vaccination, compared with dogs with current vaccination status. Most (26/33) cats, regardless of rabies vaccination status, had a titer ≥ 12 IU/mL 5 to 15 days after booster vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that dogs with out-of-date vaccination status were not inferior in their antibody response following booster rabies vaccination, compared with dogs with current vaccination status. Findings supported immediate booster vaccination followed by observation for 45 days of dogs and cats with an out-of-date vaccination status that are exposed to rabies, as is the current practice for dogs and cats with current vaccination status.

Associations of various physical and blood analysis variables with experimentally induced Mycoplasma bovis pneumonia in calves.

OBJECTIVE: To determine associations of blood analysis variables and orbit and nasal planum surface temperatures with the onset and severity of Mycoplasma bovis pneumonia in calves. Animals-28 healthy calves. PROCEDURES: Calves were challenged with M bovis (n = 24) on day 0 or not challenged (4). Blood samples were obtained for cardiac troponin I, CBC, and serum biochemical analyses on various days. Orbit and nasal planum surface temperatures were determined with infrared thermography on various days. Calves were euthanized, gross necropsies were performed, heart and lung samples were collected for histologic evaluation, and microbial cultures of lung samples were performed on day 14. Pneumonia severity was categorized as mild (< 10% lung consolidation) or moderate (≥ 10% lung consolidation). Associations between measured variables and severity of pneumonia or sample collection day were determined. RESULTS: Plasma cardiac troponin I concentration for the 28 calves was significantly higher on day 14 than it was on day 0 or 7 (least squares mean, 0.02, 0, and 0 ng/mL, respectively). No other variables changed significantly during the study. No substantial gross or histologic abnormalities were identified in cardiac muscle samples. Day 14 plasma fibrinogen concentration was significantly different between calves with mild pneumonia and those with moderate pneumonia (mean, 0.44 and 0.74 g/dL, respectively). Calves with moderate pneumonia had significantly lower least squares mean surface temperature of the dorsal aspect of the nasal planum (18.7°C) versus calves with mild pneumonia (22.9°C). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated the evaluated variables had low value for assessment of bovine respiratory disease complex in calves.

Precision and accuracy of clinical illness scores, compared with pulmonary consolidation scores, in Holstein calves with experimentally induced Mycoplasma bovis pneumonia.

OBJECTIVE: To determine the precision of a clinical illness score (CIS) system for identification of clinical signs in calves with experimentally induced Mycoplasma bovis pneumonia and to evaluate the accuracy of CISs in relation to pulmonary consolidation scores assigned at necropsy. ANIMALS: 178 Holstein bull calves that were 52 to 91 days of age at the time of pneumonia induction. PROCEDURES: 5 trials involved calves challenged with M bovis and scheduled for euthanasia and necropsy 12 to 24 days afterward. Nine veterinarian observers with various degrees of experience simultaneously assigned CISs to calves within 48 hours before necropsy. The precision of the CIS system among observers was evaluated via the Cohen κ statistic. The accuracy of each observer's CISs relative to 6 cutoffs (≥ 5%, ≥ 10%, ≥ 15%, ≥ 20%, ≥ 25%, and ≥ 30%) of percentage pulmonary consolidation was determined by comparing prenecropsy CISs with the gross pulmonary consolidation scores assigned at necropsy. Estimates for sensitivity and specificity were calculated relative to the 6 pulmonary consolidation cutoffs. RESULTS: A slight level of agreement was evident among observers (κ range, 0.10 to 0.21 for the individual trials) and overall (κ = 0.16; 95% confidence interval, 0.10 to 0.24). Median sensitivity and specificity changed with pulmonary consolidation score cutoff. Median sensitivity for all observers ranged from 81.7% to 98.9%, and median specificity ranged from 80.8% to 94.9% over all cutoff values. CONCLUSIONS AND CLINICAL RELEVANCE: Agreement among observers assigning CISs to calves was low; the accuracy of the CIS system in relation to that of pulmonary consolidation scoring varied with the severity of consolidation considered to represent bovine respiratory disease.

Effect of transportation during periods of high ambient temperature on physiologic and behavioral indices of beef heifers.

OBJECTIVE: To determine the effect of transportation during periods of high ambient temperature on physiologic and behavioral indices of beef heifers. ANIMALS: 20 heifers (mean body weight, 217.8 kg). PROCEDURES: Ten heifers were transported 518 km when the maximum ambient temperature was ≥ 32.2°C while the other 10 heifers served as untransported controls. Blood samples were collected from transported heifers at predetermined intervals during the transportation period. For all heifers, body weights, nasal and rectal temperatures, and behavioral indices were measured at predetermined intervals for 3 days after transportation. A week later, the entire process was repeated such that each group was transported twice and served as the control twice. RESULTS: Transported heifers spent more time near the hay feeder on the day of transportation, had lower nasal and rectal temperatures for 24 hours after transportation, and spent more time lying down for 2 days after transportation, compared with those indices for control heifers. Eight hours after transportation, the weight of transported heifers decreased 6%, whereas that of control heifers increased 0.6%. At 48 hours after initiation of transportation, weight, rectal temperature, and time spent at various pen locations did not differ between transported and control heifers. Cortisol concentrations were higher 4 hours after initiation of transportation, compared with those determined just prior to transportation. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated transportation during periods of high ambient temperatures caused transient changes in physiologic and behavioral indices of beef heifers.

Clinical, behavioral, and pulmonary changes in calves following inoculation with Mycoplasma bovis.

OBJECTIVE: To characterize clinical and behavioral changes in calves following inoculation with Mycoplasma bovis and evaluate relationships between those changes and pulmonary disease. ANIMALS: 22 healthy Holstein steers. PROCEDURES: 20 calves were inoculated intranasally with < 10(8) CFU or > 10(9) CFU of M bovis. Calves were assigned a clinical illness score (CIS) on a scale of 1 through 4 twice daily on the basis of severity of cough, labored breathing, and lethargy. For each calf, distance traveled and time spent near the waterer, feed bunk, or shelter were determined via a remote location monitoring device. Calves were euthanized and necropsied 22 days after inoculation. RESULTS: 13 calves became clinically ill after challenge inoculation; 3 calves were euthanized within 20 days. Among all calves, consolidation was evident in 0% to 79.9% of the lungs; extent of lung consolidation did not differ between the challenge dose groups. Distance traveled and percentages of time spent in proximity to the feed bunk and shelter were associated with CIS; calves with more severe disease traveled less distance and spent less time at the feed bunk and more time in the shelter. Distance traveled by calves was negatively associated with extent of lung consolidation (< or ≥ 10% of lungs affected); this effect was modified by trial day. CONCLUSIONS AND CLINICAL RELEVANCE: Following inoculation with M bovis, calf behavior patterns were associated with both CIS and severity of pulmonary disease. Use of behavior monitoring systems may aid in recognition of respiratory tract disease in calves.

Pathogenicity of swine influenza viruses possessing an avian or swine-origin PB2 polymerase gene evaluated in mouse and pig models.

PB2 627K is a determinant of influenza host range and contributes to the pathogenicity of human-, avian-, and mouse-adapted influenza viruses in the mouse model. Here we used mouse and pig models to analyze the contribution of a swine-origin and avian-origin PB2 carrying either 627K or 627E in the background of the classical swine H1N1 (A/Swine/Iowa/15/30; 1930) virus. The results showed PB2 627K is crucial for virulence in the mouse model, independent of whether PB2 is derived from an avian or swine influenza virus (SIV). In the pig model, PB2 627E decreases pathogenicity of the classical 1930 SIV when it contains the swine-origin PB2, but not when it possesses the avian-origin PB2. Our study suggests the pathogenicity of SIVs with different PB2 genes and mutation of codon 627 in mice does not correlate with the pathogenicity of the same SIVs in the natural host, the pig.

Escherichia coli serotype O157:H7 retention on solid surfaces and peroxide resistance is enhanced by dual-strain biofilm formation.

In a previous study we showed that an Escherichia coli O157:H7 strain that was unable to form biofilm was retained in large numbers in dual-strain biofilms formed with an E. coli O-:H4 companion strain. In this study we tested additional companion strains for their ability to retain E. coli O157:H7 strain 0475s. Companion strains producing biofilm that withstood aggressive washes were able to significantly increase serotype O157:H7 retention. Dual-strain biofilms with certain companion strains retained higher percentages of strain 0475s, and that ability was independent of biofilm total cell numbers. Tests with additional non-biofilm-forming E. coli O157:H7 strains showed that enhancement by companion strains was not unique to strain 0475s. Experiments using an E. coli companion strain with deletions of various curli and cellulose genes indicated that dual-strain biofilm formation was dependent on companion strain properties. Strain 0475s was not able to generate biofilm or persist on plastic when grown in broth with a biofilm-forming companion and separated by a 0.2 microm porous membrane, indicating a requirement for intimate contact with the companion strain. When dual-strain biofilms and planktonic cells were challenged with 5% H(2)O(2), strain 0475 showed greater survival in biofilms with certain companion strains compared to the corresponding planktonic cells. The results of this study indicate that non-biofilm-forming E. coli O157:H7 strains are retained on solid surfaces associated with biofilms generated by companion strains. However, properties other than biofilm mass enable certain companion strains to retain greater numbers of E. coli O157:H7.

The CsgA and Lpp proteins of an Escherichia coli O157:H7 strain affect HEp-2 cell invasion, motility, and biofilm formation.

In Escherichia coli O157:H7 strain ATCC 43895, a guanine-to-thymine transversion in the csgD promoter created strain 43895OR. Strain 43895OR produces an abundant extracellular matrix rich in curli fibers, forms biofilms on solid surfaces, invades cultured epithelial cells, and is more virulent in mice than strain 43895. In this study we compared the formic acid-soluble proteins expressed by strains 43895OR and 43895 using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and identified two differentially expressed proteins. A 17-kDa protein unique to strain 43895OR was identified from matrix-assisted laser desorption ionization-time of flight analysis combined with mass spectrometry (MS) and tandem MS (MS/MS) as the curli subunit encoded by csgA. A <10-kDa protein, more highly expressed in strain 43895, was identified as the Lpp lipoprotein. Mutants of strain 43895OR with disruption of lpp, csgA, or both lpp and csgA were created and tested for changes in phenotype and function. The results of this study show that both Lpp and CsgA contribute to the observed colony morphology, Congo red binding, motility, and biofilm formation. We also show that both CsgA and Lpp are required by strain 43895OR for the invasion of cultured HEp-2 cells. These studies suggest that in strain 43895OR, the murein lipoprotein Lpp indirectly regulates CsgA expression through the CpxAR system by a posttranscriptional mechanism.

Assessment of necropsy findings in sled dogs that died during Iditarod Trail sled dog races: 23 cases (1994-2006).

OBJECTIVE: To describe the character and frequency of causes of death and associated lesions in long-distance racing sled dogs. DESIGN: Retrospective case series. ANIMALS: 23 dogs. PROCEDURES: Medical records of dogs that died during or soon after competition in the Iditarod Trail sled dog races (1994 through 2006) were examined for fi ndings of gross necropsy and histologic evaluation of tissue samples. From the data, descriptive and comparative statistics were obtained. RESULTS: Recognized causes of death included aspiration of gastric contents (n = 4), aspiration pneumonia (4), acute blood loss secondary to gastric ulceration (3), and sled dog myopathy (2). A cause of death was not established for 7 dogs. Prevalent lesions among the study population included rhabdomyolysis (n = 15), enteritis (10), gastritis (10), aspiration pneumonia (8), and gastric ulceration (8). All dogs with aspiration pneumonia had concurrent gastric mucosal lesions. Subjective biventricular cardiac hypertrophy was evident in most dogs; other lesions detected frequently included centrilobular hepatic fibrosis, gastric dilatation, and mild cardiac myodegeneration and necrosis. CONCLUSIONS AND CLINICAL RELEVANCE: Unexpected death is a rare event among conditioned sled dogs during competition in endurance races. Potentially life-threatening conditions of dogs that are associated with periods of long-distance physical exertion include aspiration pneumonia, gastric mucosal lesions, and severe rhabdomyolysis. Dogs that develop clinical signs suggestive of these conditions should be excluded from strenuous activities. Epidemiologic investigations are required to clarify the risk for death associated with these lesions in dogs competing in endurance races.

Magnetic resonance imaging of the femoral head of normal dogs and dogs with avascular necrosis.

The purpose of this study was to describe the appearance of the femoral head of normal, young, small breed dogs, and dogs with avascular necrosis using low-field (0.3 T) magnetic resonance (MR) imaging. Images of the femoral heads were obtained in the dorsal plane, and included T1-weighted spin-echo, T2-weighted fast spin-echo, fast spin echo-inversion recovery, and fluid attenuated inversion recovery pulse sequences. MR imaging features of the asymptomatic femoral heads and necks included uniform high signal intensity compared with muscle on T1- and T2-weighted images. There was either uniform enhancement or no enhancement on postcontrast T1-weighted images. The MR imaging findings of dogs affected with avascular necrosis differed from those of asymptomatic dogs. Typically, the affected dogs had inhomogeneous intermediate to low-signal intensity within the femoral head and neck compared with muscle on T1-weighted images, inhomogeneous enhancement of the femoral head and/or neck on postcontrast T1-weighted images, and inhomogeneous low- to high- signal intensity within the femoral head and neck on T2-weighted images.

Differentiation of C2D macrophage cells after adoptive transfer.

C2D macrophage cells protect immunocompromised mice from experimentally induced pneumonias after intraperitoneal (i.p.) adoptive transfer. These macrophage cells are immature and display minimal activity in vitro. Therefore, we wanted to understand how adoptive transfer affected these cells. We believe that the in vivo environment affects the phenotypic and functional characteristics of macrophages that help maintain the physiological integrity of the host. To test this hypothesis, we characterized the trafficking patterns and cellular changes of the established macrophage C2D cell line after adoptive transfer. We examined phenotypic changes of the C2D macrophage cells in vivo with and without stimulation with gamma interferon (IFN-gamma). After in vivo i.p. adoptive transfer, C2D macrophage cells trafficked to the lungs, spleen, lymph nodes, and bone marrow of recipient mice. The cells were detected for as long as 2 months, and the cells expressed increased levels of CD11b, c-fms, and F4/80 on their surface, becoming more differentiated macrophages compared to cells maintained in vitro. Upon in vivo stimulation with IFN-gamma, c-fms levels decreased while Gr-1 levels increased compared to in vivo, unstimulated, phosphate-buffered saline-injected controls. These responses were independent of the genetic backgrounds of the recipient mice. These data support the hypothesis and indicate that C2D macrophage cells respond to in vivo signals that are absent during in vitro culture.

Use of computed tomography to evaluate pathologic changes in the lungs of calves with experimentally induced respiratory tract disease.

OBJECTIVE: To optimize methods for the use of computed tomography (CT) to assess pathologic changes in the lungs of calves and to determine the effect of treatment on lung consolidation. ANIMALS: 10 male Holstein calves. PROCEDURES: Calves were anesthetized to facilitate CT imaging of the thorax. After initial images were obtained, pneumonia was induced in the calves by inoculation through a bronchoscope. Two calves were used in a preliminary study to refine the inoculation dose and optimize CT images. Four calves were administered florfenicol and 4 calves were untreated control animals. Serial images were obtained 24, 48, and 72 hours after inoculation. After final images were obtained, calves were euthanized, and lung consolidation was estimated by use of lung surface area scoring and water displacement. These estimates were compared with estimated lung consolidation obtained by use of CT. RESULTS: Calves had rapid disease progression. Percentage of lung consolidation was not significantly different between treatment groups for any of the estimation methods. Results of an ANOVA of the 3 assessment methods indicated significant differences among methods. Estimates of the percentage of lung consolidation obtained by use of surface area scoring and CT correlated well, whereas water displacement estimates correlated poorly with other methods of consolidation estimation. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the correlation with other methods for estimation of lung consolidation, CT has the potential to be used to monitor disease progression in calves with experimentally induced respiratory tract disease.

Bovine alveolar macrophage neurokinin-1 and response to substance P.

In this study bovine alveolar macrophage neurokinin-1 (NK-1) and the in vitro response to substance P (SP) exposure were investigated. Bovine alveolar macrophage membrane extracts separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted using anti-NK-1 antiserum demonstrated the presence of an approximately 60kDa band. Phagocytosis of fluorescent bioparticles by SP-exposed macrophages was 39% greater than that of non-exposed macrophages (P=0.0089). Likewise, there was 28% greater TNF production by macrophages following SP exposure compared to non-exposed controls (P=0.116). These results suggest that bovine alveolar macrophages respond to SP at least in part by enhancing phagocytosis and TNF production.

Correlation of results of pulmonary computed tomography and pathologic findings in mice with Pasteurella-induced pneumonia.

OBJECTIVE: To determine correlation between results of computed tomography (CT) versus pathologic examination for determining the volume percentage of affected lung in mice experimentally infected with Pasteurella pneumotropica. ANIMALS: 30 adult mice. PROCEDURE: After helical CT scans on day 0, mice were inoculated intranasally with P. pneumotropica. Repeat CT scans were performed on days 1, 2, 3, 4, 6, 8, 10, and 13. Regions of interest (affected areas) were manually drawn on the CT images, and percentage volume of normal lung was calculated by use of 3 methods: first-day volume, largest volume, and last-day volume. Three mice were euthanatized for pathologic evaluation after each scan day. The lungs were examined with a dissection microscope, and lesion scores were assigned on the basis of percentage volume of pneumonia. Correlation coefficients comparing results of the 3 CT methods with results of gross examination were calculated. RESULTS: Lung abnormalities were detected via dissection microscopy by postinfection day 2 and via CT by days 2 or 3. Correlation coefficients for the 3 CT methods of analysis, compared with pathologic findings, were 0.7 via first-day lung volume, 0.8 via largest lung volume, and 0.8 via last-day lung volume. CONCLUSIONS AND CLINICAL RELEVANCE: Results of CT correlated well with results of dissection microscopy for estimating percentage volume of lung affected by pneumonia in mice experimentally infected with P. pneumotropica. This method may be useful for longitudinal studies of pneumonia in mice.

Toll-like receptor 4-positive macrophages protect mice from Pasteurella pneumotropica-induced pneumonia.

This study investigates Toll-like receptor 4 (TLR4)-positive macrophages in early recognition and clearance of pulmonary bacteria. TLR4 is a trans-membrane receptor that is the primary recognition molecule for lipopolysaccharide of gram-negative bacteria. The TLR4(Lps-del) mouse strains C57BL10/ScN (B10) and STOCK Abb(tm1) TLR4(Lps-del) Slc11a1(s)(B10 x C2D) are susceptible to pulmonary infections and develop pneumonia when naturally or experimentally infected by the opportunistic bacterium Pasteurella pneumotropica. Since these mice have the TLR4(Lps-del) genotype, we hypothesized that reconstitution of mice with TLR4-positive macrophages would provide resistance to this bacterium. A cultured macrophage cell line (C2D macrophages) and bone marrow cells from C2D mice were adoptively transferred to B10 and B10 x C2D mice by intraperitoneal injection. C2D macrophages increased B10 and B10 x C2D mouse resistance to P. pneumotropica. In C2D-recipient mice there was earlier transcription of tumor necrosis factor alpha and chemokines JE and macrophage inflammatory protein 2 (MIP-2) in the lungs of B10 and B10 x C2D mice, and there was earlier transcription of KC and MIP-1alpha in B10 x C2D mice. In addition, the course of inflammation following experimental Pasteurella challenge was altered in C2D recipients. C2D macrophages also protected B10 x C2D mice, which lack CD4(+) T cells. These data indicate that macrophages are critical for pulmonary immunity and can provide host resistance to P. pneumotropica. This study indicates that TLR4-positive macrophages are important for early recognition and clearance of pulmonary bacterial infections.