A study of experimental autoimmune encephalomyelitis in dogs as a disease model for canine necrotizing encephalitis

In the present study, the use of dogs with experimental autoimmune encephalomyelitis (EAE) as a disease model for necrotizing encephalitis (NE) was assessed. Twelve healthy dogs were included in this study. Canine forebrain tissues (8 g), including white and grey matter, were homogenized with 4 mL of phosphate-buffered saline for 5 min in an ice bath. The suspension was emulsified with the same volume of Freund''s complete adjuvant containing 1 mg/mL of killed Mycobacterium tuberculosis H37Ra. Under sedation, each dog was injected subcutaneously with canine brain homogenate at four sites: two in the inguinal and two in the axillary regions. A second injection (booster) was administered to all the dogs using the same procedure 7 days after the first injection. Clinical assessment, magnetic resonance imaging, cerebrospinal fluid analyses, necropsies, and histopathological and immunohistochemical examinations were performed for the dogs with EAE. Out of the 12 animals, seven (58%) developed clinically manifest EAE at various times after immunization. Characteristics of canine EAE models were very similar to canine NE, suggesting that canine EAE can be a disease model for NE in dogs.     

Carbohydrate metabolism and hepatic zonation in the Atlantic hagfish,Myxine glutinosa liver: effects of hormones

Viable Atlantic hagfish (Myxine glutinosa) hepatocytes were isolated from combined or separated large and small lobes and carbohydrate metabolism was studied. Cells had low levels of glycogen (16–30 μmol·g⁻¹), and low rates of total glucose production (TGP; 0–480 nmol·h⁻¹·g⁻¹ cells). Lactate flux to glucose (5.5 nmol·h⁻¹·g⁻¹) and CO₂ (76 nmol·h⁻¹·g⁻¹) was lower than reported values for teleosts, with a low percentage (30%) of the lactate carbon reaching glucose. Insulin significantly increased total glucose production and gluconeogenesis and decreased 6-phosphofructo 1-kinase (PFK-1) activities and glucose oxidation, while glucagon was without effect on any parameter studied. Forskolin significantly increased TGP. Epinephrine (Epi), norepinephrine (NEpi), isoproterenol (Iso), and phenylephrine (Phe) all decreased CO₂ production from lactate; propanolol blocked the effects of Epi, NEpi, and Iso. The large lobe, accounting for 65% of total liver mass, had a higher glycogen content and higher CO₂ production from lactate compared to the small lobe. Furthermore, enzyme activities in the large lobe were greater than in the small lobe, with the exception of glycogen phosphorylase (GPase) which exhibited smaller %a values in the large lobe. These data indicate the presence of a hormonally-responsive carbohydrate metabolism in hagfish hepatocytes, which is qualitatively and quantitatively different between the two liver lobes.     

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.     

Effects of dietary thyroid hormones on the red drum (Sciaenops ocellatus)

Four separate 8-week feeding trials were conducted to assess the effects of supplementing semipurified diets with either triiodothyronine (T₃) or thyroxine (T₄) at 0, 2, 10, and 50 mg/kg on growth and body composition of juvenile red drum (Sciaenops ocellatus) held in artificial brackish water (6‰) and artificial seawater (32‰). At both levels of salinity, increasing doses of T₃ resulted in fish with reduced weight gain, feed efficiency, condition factor (weight × 100/length³), and muscle ratio (muscle weight × 100/body weight), as well as a lighter body color. Significant (p < 0.05) effects of T₃ on the proximate composition of whole body, liver, and muscle were variable, generally reflecting decreased lipid and protein storage in liver and muscle, respectively. The two highest doses of T₃ given to seawater adapted fish increased survival. Dietary T₄ supplementation had no distinctive effects on appearance, growth or proximate body composition. These results indicate that whereas T₃ may function to regulate protein and lipid metabolism in red drum, dietary supplementation with T₃ leads to a hyperthyroidism-induced catabolic state. The elevated endogenous thyroid hormone levels found in fish fed optimal diets may thus adequately supply tissue needs during juvenile growth.     

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.     

Cortisol does not affect hepatic α- and β-adrenoceptor properties in rainbow trout (Oncorhynchus mykiss)

Distribution and function of hepatic - and -adrenoceptors were examined in rainbow trout (Oncorhynchus mykiss) injected with slow release hydrogenated coconut oil implants alone (sham) or containing cortisol. - and -Adrenoceptors were assayed on purified hepatic membranes 10–14 days post-implantation using ³H-prazosin () and ³H-CGP (). At 10–14 days, plasma cortisol values were significantly elevated to approximately 220 compared with 35.0 ng ml^-1 in cortisol implanted vs. sham trout. No significant differences were found between any of the experimental groups for either the affinity (K_d) or maximal number of binding sites (B_max) for either receptor type. Epinephrine significantly stimulated glucose release from hepatocytes isolated from sham injected trout, but not from cortisol-treated fish. Epinephrine-induced glucose release was blocked by both - and -antagonists. These studies do not support the hypothesis that rainbow trout exposed to chronic cortisol alter properties of hepatic adrenoceptors.     

Comparison of the width of the intervertebral disk space and radiographic changes before and after intervertebral disk fenestration in dogs

Intervertebral disk space widths were measured on lateral radiographs of 73 anesthetized dogs. Weight was found to have a significant (P less than 0.01) effect on disk space width. Using weight-adjusted disk space width measurements for all subsequent studies, older (7- to 16-year-old) dogs and males had consistently, but not significantly, wider, disk spaces than did alternative groups. Cervical and lumbar intervertebral disk spaces tended to be wider than those in the caudal thoracic region. The widest cervical intervertebral disk spaces were C4-5 and C5-6 and the narrowest was C2-3. In the lumbar region, L2-3 was the widest disk space and L4-5 was the narrowest. Dachshunds generally had greater mean intervertebral disk space width than did other breeds of dogs. Cervical (n = 6 dogs) and thoracolumbar (n = 6 dogs) disk fenestration resulted in narrow intervertebral disk spaces, regardless of breed. When a ventral approach was used in thoracolumbar fenestration, the mean intervertebral disk space was narrower than that resulting from use of a dorsolateral approach. Spondylosis was found radiographically 1 to 4 years after intervertebral disk fenestration in 3 of 6 dogs that had cervical fenestrations and in 5 of 6 dogs that underwent thoracolumbar fenestration.