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.     

Repellency of Cinnamomum cassia bark compounds and cream containing cassia oil to Aedes aegypti (Diptera: Culicidae) under laboratory and indoor conditions

Patch and skin bioassays were used in laboratory and indoor tests to evaluate the repellency of (E)-cinnamaldehyde, identified in Cinnamomum cassia Blume bark and essential oil, and a cream containing 5% (w/w) cassia oil against Aedes aegypti (L.) females. Results were compared with those of a known C. cassia compound cinnamyl alcohol, N,N-diethyl-m-toluamide (DEET) and two commercial repellents: MeiMei cream containing citronella and geranium oils and Repellan S aerosol containing 19% DEET. In patch bioassay tests with A. aegypti females, (E)-cinnamaldehyde at 0.153 mg cm(-2) and DEET at 0.051 mg cm(-2) provided 93 and 89% protection at 40 min after exposure. In skin bioassay tests, (E)-cinnamaldehyde at 0.051 mg cm(-2) and DEET at 0.025 mg cm(-2) provided 87 and 95% protection at 30 min after application. (E)-Cinnamaldehyde was significantly more effective than cinnamyl alcohol in both bioassays. In indoor tests with four human volunteers, 5% cassia oil cream provided 94, 83 and 61% protection against A. aegypti females exposed for 30, 50 and 70 min after application respectively. Cassia oil cream was a slightly less effective repellent than MeiMei cream. Repellan S aerosol provided 91% repellency at 120 min after application. Products containing cassia oil merit further study as potential repellents for the protection of humans and domestic animals from blood-feeding vectors and the diseases they transmit.     

Measurement of vitamin A,vitamin E,selenium, and L-lactate in dogs with and without osteoarthritis secondary to ruptured cranial cruciate ligament

This study compared vitamin A, vitamin E, selenium (Se), and L-lactate in blood and synovial fluid in 2 groups of 6 dogs; a control group (without OA) and an osteoarthritic group with spontaneous cranial cruciate ligament rupture and OA. Concentrations of vitamin E were significantly higher in serum than in synovial fluid in both OA (P = 0.006) and control (P = 0.0008) groups. Vitamin E concentration in synovial fluid was significantly higher in the OA group than in the control group (P = 0.009). Concentrations of Se were significantly higher in serum than in synovial fluid in both OA (P = 0.003) and control (P = 0.0006) groups. There were no significant differences in levels of Se, vitamin A, and L-lactate between the 2 groups. This is the first study to show an increased concentration of vitamin E in the synovial fluid of dogs with OA compared with dogs that did not have OA.     

Somatotropin and adipose tissue metabolism: substrate and temporal effects

The purpose of these studies was to determine the time course for changes in feed intake, blood metabolites, and lipogenic activity in adipose tissue in response to the initiation of porcine somatotropin (pST) treatment and following withdrawal from treatment in barrows. An initial study was conducted to determine the impact of chronic pST treatment (4 wk of daily injection; 0 vs 4 mg/d) on adipose tissue lipid metabolism in barrows (initial weight 67 kg). Feed efficiency was improved 27%, backfat thickness was decreased 43%, and glucose and lactate oxidation and incorporation into lipid in adipose tissue was reduced 70 to 86% in pST-treated pigs. Palmitate esterification was decreased 44%, whereas palmitate oxidation was unaffected. In vitro metabolism of lactate, glucose, and palmitate in liver slices was not affected by pST treatment. The time-course for changes in intake and adipose tissue metabolism in response to 7 d of pST (0 vs 4 mg/d) treatment and 7 d of withdrawal was examined in subsequent studies in barrows (initial weight 75 kg). Feed intake during pST treatment was significantly (P < .05) less than in control pigs within 24 h of the initiation of treatment and remained low through 3 d after withdrawal. Adipose tissue biopsies were obtained on d 0, 1, 2, 4, and 7 of the treatment phase and on d 2, 4, and 7 after withdrawal from 7 d of treatment. Maximal inhibition of lipogenesis by pST treatment in adipose tissue in vitro was observed on d 4 (-68%) and d 7 (-69%). Similarly, fatty acid synthase activity declined during the treatment period, with the greatest change noted on d 7 (-26%). After withdrawal from treatment, lipogenesis gradually increased, returning to control values 7 d after withdrawal. Levels of IGF-I began to increase from d 1 to d 7 of treatment, continually decreased during withdrawal, and were normalized by the end of the withdrawal period. Plasma urea nitrogen concentrations decreased during treatment, increased during the withdrawal phase, and were normalized 4 d after the last pST treatment. Overall results indicate that most of the metabolic changes in response to pST occur within 1 wk of treatment and return to pretreatment values after 7 d of withdrawal from treatment.     

Detection and characterization of bovine-like coronaviruses from four species of zoo ruminants

Five coronaviruses (CoVs) were detected in diarrheal feces from four zoo ruminant species: one wisent (Bison bonasus), two Himalayan tahr (Hemitragus jemlahicus), one sitatunga (Tragelaphus spekii), and one nyala (Tragelaphus angasii). We sequenced and analyzed the spike (S) and hemagglutinin/esterase (HE) genes of these viruses and compared the nucleotide (nt) and deduced amino acid (aa) sequences with those of other bovine CoV (BcoV) strains. Comparison of the entire deduced aa sequences of the S and HE glycoproteins revealed no specific differences that would account for discrimination between bovine-like CoV strains from zoo ruminants and BcoVs strains. In addition, the 99.9% aa identity among the five CoV strains revealed that the ruminants were infected by the same strain. Phylogenetically, bovine-like CoVs belong to group 2a CoVs, which are related most closely to the BcoV strains recently isolated in Korea. These data suggest that cattle are potential reservoirs for CoVs that are capable of infecting zoo ruminants.