Evaluation of host and bacterial gene modulation during Lawsonia intracellularis infection in immunocompetent C57BL/6 mouse model

BackgroundProliferative enteritis caused by Lawsonia intracellularis undermines the economic stability of the swine industry worldwide. The development of cost-effective animal models to study the pathophysiology of the disease will help develop strategies to counter this bacterium.ObjectivesThis study focused on establishing a model of gastrointestinal (GI) infection of L. intracellularis in C57BL/6 mice to evaluate the disease progression and lesions of proliferative enteropathy (PE) in murine GI tissue.MethodsWe assessed the murine mucosal and cell-mediated immune responses generated in response to inoculation with L. intracellularis.ResultsThe mice developed characteristic lesions of the disease and shed L. intracellularis in the feces following oral inoculation with 5 × l0⁷ bacteria. An increase in L. intracellularis 16s rRNA and groEL copies in the intestine of infected mice indicated intestinal dissemination of the bacteria. The C57BL/6 mice appeared capable of modulating humoral and cell-mediated immune responses to L. intracellularis infection. Notably, the expression of genes for the vitamin B12 receptor and for secreted and membrane-bound mucins were downregulated in L. intracellularis -infected mice. Furthermore, L. intracellularis colonization of the mouse intestine was confirmed by the immunohistochemistry and western blot analyses.ConclusionsThis is the first study demonstrating the contributions of bacterial chaperonin and host nutrient genes to PE using an immunocompetent mouse model. This mouse infection model may serve as a platform from which to study L. intracellularis infection and develop potential vaccination and therapeutic strategies to treat PE.     

Comparison of arylalkylamine N-acetyltransferase and melatonin receptor type 1B immunoreactivity between young adult and aged canine spinal cord

Melatonin affects diverse physiological functions through its receptor and plays an important role in the central nervous system. In the present study, we compared immunoreactivity patterns of arylalkylamine N-acetyltransferase (AANAT), an enzyme essential for melatonin synthesis, and melatonin receptor type 1B (MT2) in the spinal cord of young adult (2~3 years) and aged (10~12 years) beagle dogs using immunohistochemistry and Western blotting. AANAT-specific immunoreactivity was observed in the nuclei of spinal neurons, and was significantly increased in aged dog spinal neurons compared to young adult spinal neurons. MT2-specific immunoreactivity was found in the cytoplasm of spinal neurons, and was predominantly increased in the margin of the neuron cytoplasm in aged spinal cord compared to that in the young adult dogs. These increased levels of AANAT and MT2 immunoreactivity in aged spinal cord might be a feature of normal aging and associated with a feedback mechanism that compensates for decreased production of melatonin during aging.     

Quantification of surgical blood loss

OBJECTIVE: To compare gravimetric and colorimetric methods of quantifying surgical blood loss, and to determine if there is a correlation between preoperative hemostatic tests (buccal mucosa bleeding time [BMBT] and intraoperative blood loss). STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n=15) admitted for cutaneous tumor excision, orthopedic procedure, or exploratory laparotomy. METHODS: Intraoperative blood loss was quantified by measuring irrigation fluid and weighing surgical sponges used for blood and fluid collection during surgery. Results of gravimetric measurements were then correlated to blood loss quantified using spectrophotometric analysis of hemoglobin (Hb) content. Hemostatic variables including BMBT were measured before surgery and compared with the calculated amount of blood loss. RESULTS: Blood loss quantified by gravimetric measurement showed a significant correlation with colorimetric determination of Hb content in surgical sponges and collected irrigation fluid (r=0.93, P<.0001). BMBT correlated weakly but significantly with intraoperative blood loss (r=0.56, P<.05). CONCLUSIONS: Quantifying intraoperative blood loss using spectrophotometric Hb analysis accurately assessed the amount of blood loss; however, it is a time-consuming procedure, primarily applicable as a research tool. Gravimetric evaluation of intraoperative blood loss was found to be an accurate method, which can be recommended for use in a clinical setting. CLINICAL RELEVANCE: Estimation of blood loss using a gravimetric method is accurate and applicable in the clinical setting and provides surgeons with a simple and objective tool to evaluate intraoperative blood loss.     

The Relationship Between Body Weight,Body Condition,and Survival in Cats with Heart Failure

Background: Obese people with heart failure have improved survival compared with their normal or underweight counterparts. The purpose of this study was to determine if there is a relationship between body weight or body condition and survival in cats with heart failure. Hypothesis: Body weight and body condition score (BCS) are predictors of survival in cats with heart failure. Animals: One‐hundred and one cats with heart failure (International Small Animal Cardiac Health Council Classes II, IIIa, or IIIb) evaluated between March 2007 and June 2009. Methods: Data regarding initial body weight and BCS, subsequent changes in body weight, and treatment were collected from records and compared with survival times. Results: Median initial body weight was 5.1 kg (range, 2.2–9.5 kg). Median BCS was 5 (range, 3–9). Of the 68 cats that were discharged from the hospital, median body weight change was 0.0 kg (range, ?2.6 to +2.3 kg). Survival time for all 101 cats was 93 days (0–811 days). Survival could be predicted using a model combining initial body weight (P= .02), body weight squared (P= .02), and survival to discharge (P < .001) with a resulting global P value for this model of P < .0001. Conclusions and Clinical Importance: Cats with the lowest and highest body weights had reduced survival times compared with those with body weights in the intermediate ranges, suggesting a U‐shaped relationship between body weight and survival. Additional research into the effects of body composition could help to determine optimal management of cats with heart failure.     

A preliminary study of dietary protein requirement of juvenile marbled flounder (Pseudopleuronectes yokohamae)

An 8-week feeding trial was conducted to determine the optimal dietary protein level for juvenile marbled flounder. Five semi-purified test diets were formulated to contain different protein levels (CP) including 42.7%, 47.4%, 53.3%, 58.8%, and 64.5% (dry matter), named as CP42.7, CP47.4, CP53.3, CP58.8, and CP64.5, respectively. Five hundred and twenty-five juveniles (6.0 ± 0.1 g) were randomly distributed into 15 tanks (300 L tanks), resulting in 35 fish per tank (n = 3 tanks). Fish were fed the test diets 5 times per day until satiation. The CP58.8 resulted in the highest gain in weight and the best efficiency in feed utilization among the tested protein levels (P < 0.05). Fish fed the CP58.8 diet showed significantly higher whole-body protein and lipid contents than the fish that were fed the other diets (P < 0.05). Fish fed the CP53.3, CP58.8, and CP64.5 diets showed a significantly higher dorsal-muscle lipid content than the fish that were fed the CP42.7 and CP47.4 diets (P < 0.05). The one-slope straight broken-line regression analysis on the results of the thermal growth coefficient and feed conversion ratio indicated that the estimated optimum dietary protein level was 58.8%. Taken together, it is suggested that the dietary protein level of 58.8% is optimal for better growth and high efficiency in feed utilization for the juvenile marbled flounder.     

Alteration in intestinal morphologic features associated with extensive large-colon resection in horses

Light microscopy, morphometry, and scanning electron microscopy were used to examine the mucosal morphologic features of 7 intestinal specimens (3 from the small intestine; 4 from the large intestine) from each of 8 horses 1 year after sham operation (group 1; n = 3) or extensive large-colon resection (group 2; n = 5). Qualitative light microscopic examination did not reveal differences between groups, but morphometry revealed significantly (P less than 0.05) greater intercrypt area and distance in horses with colon resection and this was most pronounced in the cecum and remaining right ventral and dorsal colon. Crypt area and depth were similar for horses with colon resection and sham operation (P greater than 0.05). Qualitative evaluation of the scanning electron micrographs revealed more prominent crypt orifices in the large intestine of horses with colon resection. The larger intercrypt distance in the colon of horses with resection was not an obvious feature of the qualitative evaluation of the surface with scanning electron microscopy. Small intestinal morphologic features were variable and significant differences were not detected between horses with sham operation and colon resection. Horses adapted to extensive large-colon resection within 1 year by increasing the absorptive (intercrypt) surface area of the remaining large intestine.     

Application of non-structural protein ELISA kits in nationwide FMD surveillance in pigs to demonstrate virus circulation in Taiwan

Large scale surveillance of FMD non-structural protein (NSP) antibody in pigs was conducted to monitor for FMD virus circulation in Taiwan using Ceditest and UBI NSP ELISA kits after recurrence of FMD in 2009. A total of 53,759 serum samples were collected from pigs in the auction markets in 2009. There were 43 farms with positive FMD NSP reactors to both NSP ELISA tests in the nationwide surveillance. After tracing back, clinical examination and the NSP ELISA testing using both Ceditest and UBI on 14 follow-up serum samples from all the herds with confirmed NSP reactors in 2009, there were 4 farms classified as positive on follow-up testing criteria. In this surveillance, we have demonstrated that the NSP ELISA tests of outbreak farms followed by clinical and serological investigation could be used to detect FMD circulation in the pig population in Taiwan even while the national compulsory vaccination program is ongoing.     

Development of a lameness model in sheep for assessing efficacy of analgesics

Objective To develop a lameness model to assess the efficacy of analgesics for alleviating pain, swelling and systemic signs of inflammation in sheep. Procedures The response to subcutaneous injection of 0.1 or 0.2 mL turpentine in a forelimb pastern (n = 4 ewes per dose) was examined at 0, 3, 6, 24, 48 and 72 h. In a second experiment, responses were measured at 0, 2, 4, 6, 8, 10, 12 and 24 h in ewes receiving 0.1 mL turpentine ± meloxicam 1 mg/kg IV at 0 h (n = 6 per group). Responses measured included forceplate pressure, skin temperature, limb circumference, nociception, leucocyte count, neutrophil : lymphocyte ratio, haptoglobin and daily feed intake. Results Turpentine injection caused a decrease in weight borne on the treated limb, increased skin temperature, increased sensitivity at the injection site and leucocytosis by 2 h and increased limb circumference by 4 h. Weight borne and sensitivity of the injected limb returned to control levels after around 24 h, whereas tissue swelling, elevated skin temperature and elevated haptoglobin levels persisted for at least 72 h. Treatment with meloxicam improved weight borne by and tolerance to pressure exerted on the turpentine‐injected limb. Conclusions The local and systemic signs of inflammation and pain, temporary reduction in function of the affected limb and partial amelioration of some of these changes by the dose of meloxicam used here suggest that injection of turpentine in the lower forelimb provides a suitable model for examining the efficacy of analgesics for alleviation of pain and inflammation in sheep.