Loop-mediated isothermal amplification for the rapid, sensitive, and specific detection of the O9 group of Salmonella in chickens

In the present study, the loop-mediated isothermal amplification (LAMP) assay was developed to amplify the fragments of the O9 Salmonella-specific insertion element and evaluated in the laboratory for its potential use in a field situation, such as poultry farms. Among the bacteria tested, a positive reaction was observed only for 128 strains of 6 serovars of the O9 group Salmonella, such as Enteritidis (SE) and Pullorum. The detection limit of the LAMP assay was 10(3)CFU/ml, which was more sensitive than that of the polymerase chain reaction (PCR) assay with the same target gene (10(6)CFU/ml). The final results were obtained within 30 min for the LAMP assay, while the PCR assay needed a total of 120 min. When the LAMP assay was applied to the enrichment broth mixed with cecal dropping samples either spiked with SE in vitro or excreted by SE-inoculated hens, the results were comparable to those of the conventional plating method including 2 separate enrichments. In conclusion, the LAMP assay developed in the present study is an effective method for the specific detection of the O9 group Salmonella serovars, including SE.     

Preparation of Photocrosslinked Fish Elastin Polypeptide/Microfibrillated Cellulose Composite Gels with Elastic Properties for Biomaterial Applications

Photocrosslinked hydrogels reinforced by microfibrillated cellulose (MFC) were prepared from a methacrylate-functionalized fish elastin polypeptide and MFC dispersed in dimethylsulfoxide (DMSO). First, a water-soluble elastin peptide with a molecular weight of ca. 500 g/mol from the fish bulbus arteriosus was polymerized by N,N′-dicyclohexylcarbodiimide (DCC), a condensation reagent, and then modified with 2-isocyanatoethyl methacrylate (MOI) to yield a photocrosslinkable fish elastin polypeptide. The product was dissolved in DMSO and irradiated with UV light in the presence of a radical photoinitiator. We obtained hydrogels successfully by substitution of DMSO with water. The composite gel with MFC was prepared by UV irradiation of the photocrosslinkable elastin polypeptide mixed with dispersed MFC in DMSO, followed by substitution of DMSO with water. The tensile test of the composite gels revealed that the addition of MFC improved the tensile properties, and the shape of the stress–strain curve of the composite gel became more similar to the typical shape of an elastic material with an increase of MFC content. The rheology measurement showed that the elastic modulus of the composite gel increased with an increase of MFC content. The cell proliferation test on the composite gel showed no toxicity.     

INTRAOPERATIVE SPINAL ULTRASONOGRAPHY IN DOGS: NORMAL FINDINGS AND CASE-HISTORY REPORTS

Intraoperative spinal ultrasonography was performed in cervical and lumbar spine of 2 and 5 normal dogs, respectively, following ventral slot technique or dorsal or hemilamenectomy. The dura was hyperechoic, while the parenchyma was hypoechoic. The subarachnoid space was anechoic. An echogenic line was present in the center of the spinal cord, as seen in human. Pulsation of the spinal cord was noted during M-mode imaging. Clinical findings of one dog with thoracolumbar disk herniation and one with thoracic vertebral fracture/subluxation confirmed the usefulness of intraoperative spinal ultrasonography for real time evaluation of spinal canal spatial abnormalities (mass lesion and degree of spinal cord compression on scanning planes) and spinal cord motion. Follow-up ultrasound examinations were possible from 6 days postoperatively.     

Presence of surfactant proteins in the uteri and placentae of pregnant mares

Immunohistochemical investigations of the expression of surfactant protein A (SP-A) and surfactant protein D (SP-D) in the uterine and placental tissues of 13 pregnant mares were performed using anti-horse monoclonal primary antibodies. Strong positive reactions for both SP-A and SP-D were observed in the trophoblasts in the microcotyledons of the placentae at 182 to 314 days of gestation; in uterine glandular epithelial cells, faint-to-weak reactions were observed during gestation. This study describes, for the first time, the changes in the SP-A and SP-D expression levels in the endometrium of mares during gestation; the SP-A and SP-D expression levels increased after the second trimester of gestation.     

Juvenile bird acanthocephalans recovered incidentally from raccoon dogs (Nyctereutes procyonoides viverrinus) on Yakushima Island, Japan

Helminth parasites were collected from 14 raccoon dogs (Nyctereutes procyonoides viverrinus) and 2 Japanese weasels (Mustela itatsi sho) on Yakushima Island, Kagoshima Prefecture (Kyushu), Japan. In addition to trematodes and nematodes reported separately, everted cystacanths of 3 bird acanthocephalans (Mediorhynchus robustus in 5 animals, Porrorchis leibyi in 2 animals, and Sphaerirostris lanceoides in 1 animal) were collected from the raccoon dogs introduced recently on this World Natural Heritage Area. A cystacanth of P. leibyi was collected also from a weasel indigenous to this island that were killed by traffic accidents. Recovery of everted cystacanths of M. robustus from the intestine of raccoon dogs was remarkably frequent (36%), suggesting that this acanthocephalan should be highly prevalent in passerine hosts on Yakushima Island, and the animals might vigorously take insect hosts or paratenic hosts. Furthermore, the present report is the second on M. robustus after its first record in Japan (Aichi Prefecture, Honshu) as M. garruli from a Eurasian jay (Garrulus glandarius japonicus) by Yamaguti in 1939. Porrorchis leibyi has never been recorded in Japan.     

Usefulness of hemilaminectomy for cervical intervertebral disk disease in small dogs

Hemilaminectomy was performed to treat cervical disc disease in 18 small dogs. Cervical spinal cord compression was characterized by ventral and/or lateral compression on myelograms. The duration of follow-up examinations ranged from 2 to 72 months. The optimal response time after surgery ranged from 2 days to 3 months. The outcome was determined to be excellent if clinical signs resolved and the dog had completely improved. The outcome was determined to be good if the dog improved, but was not clinically normal or if the degree of the owner's satisfaction was insufficient. Fourteen dogs achieved complete neurologic recovery without complications. One dog was initially neurologically worse after surgery, but ultimately improved to normal neurologic status. These outcomes were judged to be excellent. In the remaining 3 dogs, 2 dogs had relapse of neck pain and one dog remained mildly ataxic. These outcomes were judged to be good. These results suggest that hemilaminectomy is an effective option for surgical treatment of spinal cord compression secondary to cervical disc disease in small dogs.     

Clinical and clinico-pathologic characteristics of Shiba dogs with a deficiency of lysosomal acid beta-galactosidase: a canine model of human GM1 gangliosidosis

The present study was conducted to determine the clinical and clinico-pathologic characteristics of Shiba dogs with GM1 gangliosidosis, which is due to an autosomal recessively inherited deficiency of lysosomal acid beta-galactosidase activity. Clinical and clinico-pathological features were investigated in 10 homozygous Shiba dogs with GM1 gangliosidosis. The age at onset was 5 to 6 months and the dogs manifested progressive neurologic signs including loss of balance, intermittent lameness, ataxia, dysmetria and intention tremor of the head. The dogs were unable to stand by 10 months of age due to a progression of ataxia and spasticity in all limbs. Corneal clouding, a visual defect, generalized muscle rigospasticity, emotional disorder and a tendency to be lethargic were observed at 9 to 12 months. The dogs became lethargic from 13 months of age. The survival period seemed to be 14 to 15 months. As a clinico-pathologic feature, lymphocytes with abnormally large vacuoles were observed in peripheral blood (30 to 50% of total lymphocytes) through the lifetime of the dogs. The clinical and clinico-pathologic characteristics of this animal model are useful for not only the development and testing of potential methods of therapy, but also the diagnosis of affected homozygous Shiba dogs in veterinary clinics.     

Effects of dietary lysine/protein ratio and fat levels on growth performance and meat quality of finishing pigs

This study aimed to evaluate the effects of dietary lysine/protein ratio and fat levels on the growth, carcass characteristics and meat quality of finishing pigs fed feed made from food waste, including noodles and chocolate. Four dietary treatments, 2 levels of lysine/protein ratio (0.035 and 0.046) and 2 levels of fat (3.3% and 6.0%), were adapted to a 2 × 2 factorial arrangement. Each diet for the finishing pigs contained the same levels of adequate crude protein (16%) and lysine (0.58–0.75%), and similar levels of high total digestible nutrients (90.2–92.6%). In total, 32 LWD pigs with an average body weight of 57.2 kg were assigned to 4 dietary groups. The pigs were slaughtered at about 115 kg. Growth performance was not influenced by the dietary treatments. Carcass characteristics were slightly influenced by the dietary fat level. As the dietary lysine/protein ratio decreased, the marbling score of Longissimus dorsi muscle increased and the intramuscular fat (IMF) increased from 6.82% to 9.46%. Marbling score was not significantly influenced by the dietary fat level. These results indicate that IMF increased without adverse effects on growth, carcass characteristics and meat quality, when pigs were fed a diet with low lysine/protein ratio.