Two Korean black goat (approx. 2 and 3 years old) showing diarrhea and chronic weight loss were submitted to Animal and Plant Quarantine Agency. At necropsy, there were thickening of small intestine and enlargement of mesenteric lymph nodes. Microscopically, they had granulomatous enteritis in the small and large intestine and granulomatous lymphadenitis. By polymerase chain reaction (PCR) and acid fast stain, strong positive reaction and acid-fast rod bacteria were detected. According to the result of histopathology and PCR, we confirmed

The attenuated Salmonella typhimurium χ4550 strain was used to harbour a reconstructed bicistronic DNA vaccine against porcine rotavirus, which carried the rotavirus nonstructural protein 4 (NSP4) and VP7 genes simultaneously. Using a balanced lethal system, the kanamycin resistance gene of expressing eukaryotic plasmids pVAX1 and pVAXD were replaced by the aspartate β-semialdehyde dehydrogenase (asd) gene. The NSP4 cleavage product (259-525) of rotavirus OSU strain and VP7 full-length genes were amplified by reverse transcription polymerase chain reaction and then inserted into the eukaryotic single-expression plasmid, pVAX1-asd, and the eukaryotic dual-expression plasmid, pVAXD-asd, respectively. The recombinant plasmids pVAX1-asd-NSP4, pVAX1-asd-VP7 and pVAXD-asd-NSP4-VP7 were transformed into the attenuated S. typhimurium χ4550 strain by electrotransformation. An indirect immunofluorescence assay of the expressed COS-7 cell suggested that the recombinant S. typhimurium χ4550 strain was constructed successfully. The recombinant S. typhimurium χ4550 strain was orally administered to BALB/c mice. The group immunised with dual-expression plasmids produced a significantly higher level of serum Immunoglobulin G (IgG) and intestinal Immunoglobulin A (IgA) than the group immunised with single-expression plasmids. These results indicated that eukaryotic bicistronic plasmid DNA vaccines could be successfully constructed to enhance humoural, mucosal and cellular immune response against rotavirus infection.

The apxIC genes of the Actinobacillus pleuropneumoniae serovar 5 (SC-1), encoding the ApxI-activating proteins, was deleted by a method involving sucrose counter-selection. In this study, a mutant strain of A. pleuropneumoniae (SC-1) was constructed and named AapxIC/ ompP2. The mutant strain contained foreign DNA in the deletion site of ompP2 gene of Haemophilus parasuis. It showed no haemolytic activity and lower virulence of cytotoxicity in mice compared with the parent strain, and its safety and immunogenicity were also evaluated in mice. The LD50 data shown that the mutant strain was attenuated 30-fold, compared with the parent strain (LD50 of the mutant strain and parent strain in mice were determined to be 1.0 x 10(7) CFU and 3.5 x 10(5) CFU respectively). The mutant strain that was attenuated could secrete inactivated ApxIA RTX toxins with complete antigenicity and could be used as a candidate live vaccine strain against infections of A. pleuropneumoniae and H. parasuis.

In order to get a reliable estimate of brucellosis prevalence in Eritrean dairy cattle, a cross- | sectional study was carried out in 2009. The survey considered the sub-population of dairy cattle reared in modern small- and medium-sized farms. Samples were screened with the Rose Bengal test (RBT) and positive cases were confirmed with the complement fixation test (CFT). A total of 2.77% (417/15 049; Credibility Interval CI: 2.52% - 3.05%) of the animals tested in this study were positive for antibodies to Brucella species, with a variable and generally I: low distribution of positive animals at regional level. The highest seroprevalence was found in the Maekel region (5.15%; CI: 4.58% - 5.80%), followed by the Debub (1.99%; CI: 1.59% -2.50%) and Gash-Barka (1.71%; CI: 1.34% - 2.20%) regions. Seroprevalence at sub-regional levels was also generally low, except for two sub-regions of Debub and the sub-region Haicota I: from the Gash-Barka region. Seroprevalence was high and more uniformly distributed in the Maekel region, namely in the Asmara, Berik and Serejeka sub-regions. Considering the overall low brucellosis prevalence in the country, as identified by the present study, a brucellosis I: eradication programme for dairy farms using a test-and-slaughter policy would be possible. However, to encourage the voluntary participation of farmers to the programme and to raise their awareness of the risks related to the disease for animals and humans, an extensive public awareness campaign should be carefully considered, as well as strict and mandatory dairy movement control.

The indirect effects of mastitis treatment are often overlooked in cost-benefit analyses, but it | may be beneficial for the dairy industry to consider them. The cost of mastitis treatment may increase when the duration of intra-mammary infections are prolonged due to misdiagnosis of host-adapted mastitis. Laboratory diagnosis of mastitis can be costly and time consuming, therefore cow-side tests such as the California Milk Cell Test (CMCT) and Milk Electrical Resistance (MER) need to be utilised to their full potential. The aim of this study was to determine the relative benefit of using these two tests separately and in parallel. This was done using a partial-budget analysis and a cost-benefit model to estimate the benefits and costs of each respective test and the parallel combination thereof. Quarter milk samples (n = 1860) were taken from eight different dairy herds in South Africa. Milk samples were I evaluated by means of the CMCT, hand-held MER meter and cyto-microbiological laboratory analysis. After determining the most appropriate cut-off points for the two cow-side tests, the sensitivity and specificity of the CMCT (Se = 1.00, Sp = 0.66), MER (Se = 0.92, Sp = 0.62) and the tests done in parallel (Se = 1.00, Sp = 0.87) were calculated. The input data that were used for partial-budget analysis and in the cost-benefit model were based on South African figures at the time of the study, and on literature. The total estimated financial benefit of I correct diagnosis of host-adapted mastitis per cow for the CMCT, MER and the tests done in parallel was R898.73, R518.70 and R1064.67 respectively. This involved taking the expected benefit of a correct test result per cow, the expected cost of an error per cow and the cost of the test into account. The CMCT was shown to be 11% more beneficial than the MER test, whilst using the tests in parallel was shown to be the most beneficial method for evaluating the I mastitis-control programme. Therefore, it is recommended that the combined tests should be used strategically in practice to monitor udder health and promote a pro-active udder health approach when dealing with host-adapted pathogens.

A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulata in sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T.annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.

Bovine tuberculosis is an important zoonosis and accurate diagnosis is important for its surveillance. Post-mortem diagnosis may, however, be compromised by lesions caused by other pathogens. In an investigation on its prevalence in slaughter cattle in Kenya, Mycobacterium bovis and dimorphic fungi were inadvertently identified separately or concurrently in tuberculous lesions. Beef carcasses were inspected for lesions in two abattoirs in Nairobi. Tissues with lesions were collected and transported to the laboratory. Smears of lesions were stained by acid-fast procedure and examined microscopically. Lesions were cultured in Löwenstein-Jensen (LJ) and in BBL™ Mycobacterium growth indicator tubes (MGIT) media. Mycobacteria isolates in LJ medium were identified by DNA typing. Smears of BBL TM MGIT cultures were acid-fast stained and examined microscopically. Tissue sections were stained with periodic acid-Schiff reagent before examination. Of the 929 carcasses examined, 176 had granulomatous lesions. Dimorphic fungi were detected as acid-fast negative cells in 58 (32.9; 33.5%) of the lesion smears, either alone (29.0; 16.4%) or concurrently with acid-fast bacilli (29.0; 16.4%). The fungi were also detected in some BBL TM MGIT-culture smears and lesioned tissue sections. The fungi were identified, by means of cellular morphology, as Paracoccidioides brasiliensis and Blastomyces dermatitidis. A total of 64 isolates of mycobacteria were recovered in LJ medium, 19 of which were identified as M. bovis. The present report documents native P. brasiliensis infections outside the presumed endemic region and B. dermatitidis infections in a livestock animal. The findings further indicate the importance of dimorphic fungi as a differential diagnosis of bovine tuberculosis in the region.

Objective-To characterize the ultrasonographic appearance of the canine esophagus. Animals-14 healthy Beagles. Procedures-Endoscopic ultrasonography (EUS) examinations were performed with a radial ultrasonographic gastrovideoscope in anesthetized dogs. Images were obtained at 3-cm intervals along the esophageal length to allow evaluation of the esophageal wall. Images were obtained with the probe in direct contact with the esophageal wall and with a water-filled balloon as a standoff. Results-Images were obtained with (12 dogs) and without (10) the water-filled balloon. Median thickness of the esophageal wall was 2.19 mm (range, 1.03 to 5.62 mm) in the proximal third of the esophagus, 2.15 mm (range, 1.10 to 4.45 mm) in the middle third, and 2.84 mm (range, 1.35 to 5.92 mm) in the distal third. Wall thickness differed significantly between proximal and distal thirds. Results were similar when the water-filled balloon was used. Esophageal wall layers appeared as 5 alternating hyperechoic and hypoechoic bands that could not be consistently identified in all dogs. All layers could be identified in 26 of 198 (13%) images, 3 layers could be identified in 67 of 198 (34%) images, and 105 of 198 (53%) images had no layers. Visual identification of layers in images obtained with and without the balloon did not differ significantly. Conclusions and Clinical Relevance-EUS appeared to be a useful technique for assessing esophageal wall integrity in dogs; however, complete evaluation of all layers could not be accomplished in all instances. Further studies with this technique in dogs are needed.

Objective-To assess pharmacokinetics of pregabalin in horses after a single intragastric or IV dose. Animals-5 healthy adult mares. Procedures-Horses received 1 dose of pregabalin (approx 4 mg/kg) via nasogastric tube in a crossover-design study; after a 3-week washout period, the same dose was administered IV. Food was not withheld. Plasma pregabalin concentrations in samples obtained 0 to 36 hours after administration were measured by use of ultra-performance liquid chromatography with triple quadrupole tandem mass spectrometry. Pharmacokinetic variables were estimated by means of noncompartmental analysis. Results-Mild sedation was observed in 2 horses following intragastric and IV pregabalin administration. Signs of mild, transient colic or behavioral abnormalities were observed in all horses following IV administration. After intragastric administration, median (range) maximal plasma concentration was 5.0 μg/mL (4.4 to 6.7 μg/mL), time to maximal plasma concentration was 1. 0 hour (0.5 to 2.0 hours), elimination half-life was 8.0 hours (6.2 to 9.4 hours), and area under the curve from time 0 to infinity (AUC(0-∞)) was 47.2 μg·h/mL (36.4 to 58.4 μg·h/mL). After IV administration, initial concentration was 22.2 μg/mL (19.8 to 27.7 μg/mL), elimination half-life was 7.74 hours (6.94 to 8.17 hours), and AUC0-∞ was 48.3 μg·h/mL (44.8 to 57.2 μg·h/mL). Bioavailability was 97.7% (80.7% to 109.8%). Median predicted values for minimal, mean, and maximal steady-state plasma concentrations after intragastric administration assuming an 8-hour dosing interval were 3.9, 5.3, and 6.3 μg/mL, respectively. Conclusions and Clinical Relevance-At a simulated intragastric dosage of approximately 4 mg/kg every 8 hours, median pregabalin steady-state plasma concentration in healthy horses was within the therapeutic range reported for humans. Therapeutic concentrations and safety of this dosage have not been established in horses.