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The red-crowned crane Grus japonensis in Hokkaido, Japan forms a closed population as a residence that is independent of the mainland population. Based on observations of a limited number of individuals as well as cranes in captivity, red-crowned cranes are omnivores and eat fish, worms, insects and plants in their own territories except in winter, when they are fed with dent corn that is supplied in eastern Hokkaido. DNA metabarcoding based on high throughput sequencing was carried out using universal primer sets for cytochrome oxidase subunit I gene. Feces from 27 chicks collected in June and July in the period from 2016 to 2018 and intestinal contents from 33 adult and subadult cranes that were found dead almost throughout year in 2006-2013 in the field in eastern Hokkaido were used. Although compositions varied considerably in the cranes, both insects and fish were found in adults and subadults to the same extents, while insects were predominant in chicks. Both insects and fish were detected in all seasons for adults and subadults. Horse flies, scarab beetles and weevils accounted for the most of the insects regardless of the life stage. Dace, stickleback, flatfish and sculpin were the major fish species in adults, while chicks ate almost only stickleback. The results provide the first comprehensive data on carnivorous diets in wild red-crowned cranes in eastern Hokkaido as basis for conservation of red-crowned cranes, for which the life style and area continue to change.

An unknown species of the genus Notocotylus (Digenea: Notocotylidae) was found as the larval stage from the lymnaeid snail, Radix auricularia, in a static water area of the Chubetsu River, Hokkaido, the northernmost island of Japan. A DNA barcoding identification system was applied to detect the adult stage. Through the inspection of anatid game birds in Hokkaido, Anas crecca, Anas platyrhynchos, Anas zonorhyncha, and Mareca penelope were demonstrated to serve as the definitive hosts. The detailed morphological features of the species were characterized using adults raised experimentally in immunosuppressed mice and naturally developed larvae in R. auricularia. Although the species is morphologically similar to Notocotylus attenuatus and Notocotylus magniovatus in both adult and larval stages, its taxonomic independence was confirmed by a comprehensive study based on molecular phylogeny, morphology, and ecology. Here we propose Notocotylus ikutai n. sp. for this species. The migratory behavior of the anatid hosts and the North-Eurasian distribution of R. auricularia suggest that the new species is widely distributed in the northern Far East.

The red-crowned crane ( Grus japonensis ) from eastern Hokkaido is classified as a Special Natural Monument in Japan. In this study, we determined the concentrations of persistent organic pollutants (POPs) in red-crowned crane muscle tissues (n = 47). Polychlorinated biphenyls (PCBs) had the highest median concentration (240 ng/g lipid weight), followed by dichlorodiphenyltrichloroethane and its metabolites (DDTs) (150 ng/g lipid weight), chlordane-related compounds (CHLs) (36 ng/g lipid weight), hexachlorobenzene (HCB) (16 ng/g lipid weight), hexachlorocyclohexanes (HCHs) (4.4 ng/g lipid weight), polybrominated diphenyl ethers (PBDEs) (1.8 ng/g lipid weight), and finally, Mirex (1.5 ng/g lipid weight). Additionally, a positive correlation was found among POP concentrations. No sex differences beyond body parameters were observed. Additionally, red-crowned cranes exhibited a high enantiomeric excess of (+)- alpha -HCH, with enantiomer fractions varying from 0.51 to 0.87 (average: 0.69).

The endangered red-crowned crane (Grus japonensis) is a protected species in eastern Hokkaido and injured specimens are treated with medication. The present study aimed at understanding the expression profiles of cytochrome P450 (CYP) 1-3 genes in red-crowned crane tissues. We used 14 individuals found dead in the wild in eastern Hokkaido or in Kushiro City Zoo. Nine CYP1-3 genes expressed in the liver of the red-crowned crane were identified by high-throughput sequencing, and phylogenetically classified as CYP1A5, CYP2C23, CYP2C45, CYP2D49, CYP2G19, CYP2U1, CYP2AC1, CYP3A37, and CYP3A80. Based on the quantitative real-time PCR of 13 samples, the rank order of their median expression levels was as follows: CYP3A37 > CYP2AC1 > CYP2C45 > CYP2D49 > CYP2G19 > CYP1A5 > CYP3A80 > CYP2C23. The tissue distribution of the CYP transcripts indicated that many of the CYP1–3 genes examined were mainly expressed in the tissues where drug metabolism occurs, such as the liver, kidneys, and lungs. We found that CYP3A37 was dominant at the transcript level in the liver, indicating it might play a crucial role in liver physiology and xenobiotic metabolism. Similarly, an “orphan” CYP2AC1 was expressed at relatively high levels in the kidneys and liver, suggesting a possible role in renal and liver physiology and xenobiotic metabolism. Our results establish a foundation for future studies on red-crowned cranes aiming to further understand drug sensitivity and develop medication protocols, but also contribute to national and local projects for the conservation of red-crowned crane.

A summary of avian ticks and mites from an epidemiological survey performed by the Wild Animal Medical Center of the Graduate School of Veterinary Medicine of Rakuno Gakuen University is reported, with errata for mite taxa. Ten taxa were recorded, and their taxonomic positions are shown in a table.

A helminthological survey was performed on 75 individuals of Tancho, Grus japonensis, from Hokkaido, Japan. A total of seven helminth species, including five nematode (Baruscapillaira sp., Contracaecum sp., Paracuaria adunca, Syncuaria sp., Viktorocara sp.) and two trematode (Echinostoma gotoi, Apatemon gracilis) species were obtained from 57.3% of the cranes, and E. gotoi was the most common encountered. Among the above helminths, all helminths except for E. gotoi were the first host records for G. japonensis worldwide, and Syncuaria sp. was the first geographical record from Japan. Since the acuariid nematodes including the genera Paracuaria, Syncuaria and Viktorocara are known to cause ulcerative ventriculitis and/or enteritis in bird alimentary tracts, monitoring of helminths is recommended in this endangered crane species. 2000年から2009年の間に北海道内各地で傷病個体として回収された後、斃死したタンチョウ75個体の寄生蠕虫類保有状況を調査した。57.3%から線虫5種(Baruscapillaira sp., Contracaecum sp., Paracuaria adunca, Syncuaria sp., Viktorocara sp.)および吸虫2種(Echinostoma gotoi, Apatemon gracilis)の計7種の蠕虫類のいずれかが検出された。E. gotoiを除く6種はタンチョウから初記録であり,Syncuaria sp.は日本初記録であった。得られた蠕虫類の中にはParacuada属, Syncuaria属およびViktorocara属など,消化管潰瘍や腸炎の原因となる線虫が含まれたため,絶滅危惧種であるタンチョウの保護管理上,今後寄生虫のモニタリングが必要であると考えられた。

A parasitological survey was performed on 11 species of birds introduced into Japan: Cygnus olor, Branta canadensis moffitti, Cairina moschata domestica, Phasianus colchicus karpowi, Bambusicola thoracica, Pavo cristatus, Psittacula krameri manillensis, Pycnonotus sinensis, Garrulax canorus, Leiothrix lutea, Lonchura malacca. A total of 17 taxa of external and internal parasites, including one protozoan (Eimeria sp.), five nematodes (Eucoleus perforans, Amidostomum anseris, Pseudaspidodera pavonis, Heterakis gallinarum and Synhimantus (Dispharynx) nasuta), four trematodes (Echinochasmus sp., Amphimerus anatis, Tanaisia sp. and Dicrocoeliidae gen. sp.), one cestode (unidentified) and six arthropods (Goniodes pavonis, Lipeurus maculosus, Ixodes turdus, Haemaphysalis flava, Leptotrombidium scutellare and Mouchetia sp.) were obtained and identified. Among the obtained parasites, Dicrocoeliidae gen. sp., I. turdus and L. scutellare from L. lutea, and Tanaisia sp. and L. scutellare from G. canorus, are the first host records, while P. pavonis and G. pavonis are the first geographical records from Japan.

Gastrointesitnal helminths of four age groups of Aigamo ducks, a cross between wild ducks (Anas platyrhynchos) and domestic ducks (Anas platyrhynchos domesticus), were investigated in Hokkaido, Japan. A total of five helminth species, namely Amidostomum acutum, Echinostoma revolutum, Aploparaksis sp., Hymenolepis sp., and Fimbriaria sp. were obtained and identified from 37.84% of 37 individuals of Aigamo duck. Among the above, A.acutum, Aploparaksis sp. and Hymenolepis sp. are the first host record from Aigamo ducks in Japan. And the present result suggested that the parasites, including high pathogenic and/or zoonotic species, passed from wild ducks to Aigamo ducks at outfield rearing period.

2007年12月から2010年9月に川崎市夢見ヶ崎動物公園において53種の展示動物を対象に糞便検査を実施し，寄生原虫類および蠕虫類の保有状況を調査した。糞便検査では約半数の動物種からコクシジウムのオーシストあるいは線虫卵が検出された。 また，駆虫薬イベルメクチンを投与したヘラジカ（Alces alces）の糞便から羊鞭虫Trichuris ovisが検出された。

Parasitic nematodes of 176 individuals of 15 bird species belonging to the order Anseriformes from Hokkaido, Japan were investigated. A total of 12 nematode species were obtained, namely Amidostomum anseris, A. acutum, Epomidiostomum crami, E. uncinatum, Tetrameres fissispina, Eucoleus contortus, Capillaria anatis, Baruscapillaria mergi, Contracaecum rudolphii, Echinuria uncinata, Streptocara crassicauda and Sarconema eurycerca. Among these, E. uncinatum (hosts: Anas platyrhynchos, A. poecilorhyncha, A. acuta, Mel. nigra) and E. crami (hosts: Anser albifroms, Ans. fabalis, C. cygnus, C. columbianus) were the first geographical records in Japan. There appeared to be strict host-parasite relationships between the wild swans/geese and A. anseris/E. crami, and between wild duck species and A. acutum/E. uncinatum, respectively.

Thirty-eight individuals of the Crested Serpent Eagle, Spilornis cheela perplexus Swann, 1922 collected between 2001 and 2009 from Yaeyama Archipelago, Okinawa, Japan, were investigated. A total of 11 helminth and two arthropod parasites, including seven nematodes (Eucoleus contortus, Baruscapillaria falconis, Aonchotheca caudinflata, Porrocaecum angusticolle, Procyrnea leptoptera, Microtetrameres accipiter and Synhimantus sp.), three trematodes (Basantisia ridwani, Strigea microbursa and Neodiplostomum georgesduboisi), an acanthocephalan (Centrorhynchus spilornae), a chewing louse (Craspedorrhynchus sp.) and a feather mite (Artamacarus sp.), were identified. Among these parasites, E. contortus, B. falconis, A. caudinflata, P. leptoptera, M. accipiter, Synhimantus sp. and Artamacarus sp. were the first host records for S. cheela, and all helminths and arthropods are the first host records from S. c. perplexus. With the exception of E. contortus, all helminths and arthropods were the first local records from the Yaeyama Archipelago.

To clarify the cellular immune system in dairy cows with inflammatory diseases after calving, the leukocyte subsets were examined in Holstein dairy cows. Twenty Holstein cows reared in one herd, were used in this study. Nine cows (Group 1) experienced onset of mastitis or puerperal fever within 2 weeks after calving, and the other eleven cows remained healthy (Group 2) after calving. The numbers of CD3(+), CD4(+) and CD8(+) cells tended to be lower in Group 1 than in Group 2 from the day of calving through week 1. These results suggested that the cows with inflammatory diseases might have experienced a decline in T cells by the day of calving, before the onset of disease.

Specific identification of ehrlichiae in the tissues and determination of their distribution is difficult. In this study, an in-situ hybridization method was developed to detect ehrlichial 16S rRNA in tissue specimens from mice experimentally infected with the HF strain. This strain is closely related to Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis. HF strain-specific 16S rRNA was detected in endothelial cells and monocyte-macrophages in the liver, lungs, bone marrow, spleen, lymph nodes, and large and small intestinal tissues. The results suggest that the in-situ hybridization method with a digoxigenin-labelled RNA probe specific to ehrlichial 16S rRNA will be useful for post-mortem diagnosis and for the histopathological investigation of ehrlichial infection.

As one of ecological surveys of alien avian species in Japan, internal and external parasites of 4 individuals of Hwamei Garrulax canorus and 4 individuals of Red-billed Leiothrix Leiothrix lutea collected in Kanagawa, Tokyo, and Fukuoka Prefs., Japan during from July 1999to November 2001, were investigated. Ornithoica bistativa, Ornithomya avicularia cf. aobatoensis, Haemaphysalis flava and Centrorhynchus turdi were obtained from Hwamei, and Ornithonyssus sylvialum and Anonchotaenia sp, were obtained from the Leiothrixes, respectively. All of these external parasites and the acanthocephalan species are newly recorded from these two avian species in Japan.

Cellular activation and functional cell surface markers were evaluated during experimentally-induced endotoxemia in healthy horses. Eight healthy adult horses were infused a low dose of endotoxin (lipopolysaccharide from Escherichia coli O26: B6, 30 ng/kg of body weight, IV) and five control horses were given an equivalent volume of sterile saline solution. Venous blood samples were collected for flow cytometric analysis of peripheral blood mononuclear cells (PBMCs) and to measure plasma endotoxin concentrations. Clinical signs of endotoxemia were recorded at 10, 20, 30, 40, 50 min, 1, 2, 3, 4, 8, 16, 24 and 48 hr after endotoxin or saline solution administration. Clinical findings characteristic of endotoxemia (tachycardia, tachypnea, increased rectal temperature, and leukopenia) occurred transiently in all horses administered endotoxin; however, plasma endotoxin concentrations were detectable in only 50% (4/8) of the endotoxin-infused horses. The percentage of CD4(+), CD5(+), and CD8(+) cells decreased while the percentage of CD14(+), IgM(+), and MHC class II(+) cells increased significantly after endotoxin infusion. Alterations in the immunophenotype of PBMCs from horses with experimentally-induced endotoxemia were associated with changes in vital signs, indicating that endotoxin altered the immuno balance.

To clarify the cellular immune condition in Japanese Black (JB) calves with a weak syndrome, peripheral leukocyte populations were analyzed by flow cytometry. Twenty JB calves were divided into two groups based on clinical observations; one group of calves was weak, because they had experienced an onset of diarrhea within 3 days after birth and needed treatment (Group 1 ;n=10), and the other group of calves was healthy (Group 2; n=10). With regard to leukocyte populations, CD8(+) cells and gamma delta T cells in Group 1 were markedly lower than those in Group 2 during the experimental periods. It is possible that immune-insufficiency might be based on T lymphocyte function in weak syndrome JB calves during the growth process.

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, α-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.

To determine the immunological response in lactating dairy cows infected with Salmonella (S.) Takoradi, the relationships among distributions of peripheral blood mononuclear cell (PBMC) subpopulations, endotoxin concentrations and dynamics of inflammatory cytokines in blood were investigated. The ratio of CD4+ T cells to CD8+ T cells was significantly lower in the affected cattle than in the control cattle (p0.05) to decrease in the number of CD4+ T cells in the infected cattle. In contrast, the numbers of gammadeltaT cells, MHC class II-positive cells were significantly higher in the affected cattle than in the control cattle (p0.01 respectively). Endotoxemia was found in all but one of the affected cattle. Serum IL-1 and IL-6 bioactivities were significantly higher in the affected cattle than in the control cattle (IL-1, p0.05; IL-6. p0.01). Serum TNF-alpha activities and levels were not detected in the control and affected cattle. The activities of proinflammatory cytokines determined by the bioassay are important to the relationships between concentration of endotoxin, cytokines and clinical signs. such as leukocytosis, leukopenia, fever or bacterial shedding. Serum IL-2 levels were lower in the affected cattle than in the control cattle. Serum IFN-y was not detected in the affected cattle except one. These results by the ELISA seemed to reflect the condition of subpopulation in the PBMCs from the shedding cattle. The present results suggest that cellular immunity is suppressed while the humoral immunity is activated in acute bovine salmonellosis.

A 13-year-old female mongrel dog had a pleomorphic adenoma of the lacrimal gland in the right upper orbit. The tumor measured 3.8 X 3.0 X 3.3 cm, appeared white, round, and firm, and pressed the right globe and surrounding tissues. Histopathologically, the tumor had a thin connective tissue capsule and was composed of tubules with two cell types, some resembling luminal epithelial cells making up the tubular structures and the other of myoepithelial cells. Epithelial tubules were disposed in an adenomatous fashion and separated from each other by proliferating pleomorphic myoepithelial cells. Immunohistochemically, large numbers of the luminal epithelial cells revealed an immunopositive reaction against keratin/cytokeratin (AE1/AE3), and some epithelial cells reacted against cytokeratin 14. Spindle-shaped myoepithelial cells revealed an immunopositive reaction against cytokeratin 14, α-smooth muscle actin, and vimentin. A small number of myoepithelial cells reacted against desmin. S-100 protein immunopositivity was frequently found in luminal epithelial cells and rarely in the pleomorphic myoepithelial cells. Glial fibrillary acidic protein positivity was commonly found in myoepithelial cells, myxoid matrices, and intracystic materials, but not in luminal epithelial cells.

Histopathologic and immunohistochemical examinations were performed on localized amyloidosis associated with mammary tumors in two dogs. These tumors were identified as adenoma and adenocarcinoma. An acellular, amorphous pale eosinophilic material (amyloid) was observed in the lumina of acini lined by neoplastic cells and in the stroma of the tumors. Concentrically laminated pale eosinophilic bodies (corpora amylacea) were also found in the lumina of the acini. Amyloid and corpora amylacea stained positively with Congo red with and without 5% potassium permanganate pretreatment and revealed a green birefringence under polarized light. Corpora amylacea showed an occasional Maltese-cross pattern. Immunohistochemically, amyloid and corpora amylacea usually stained positively with anti-bovine α-casein antibody but negatively with anti-human amyloid AA, anti-bovine κ-light and λ-light chains, anti-human lactoferrin, anti-human transferrin, anti-human secretory component, and anti-human polyglucosan antibodies. These findings suggested that the amyloid deposition in these canine mammary tumors was related to lactating casein.

Histological and immunohistochemical studies were carried out on the pancreas of twelve cattle of insulin-dependent diabetes mellitus (IDDM). They showed clinical signs such as persistent hyperglycemia, glycosuria and decreased glucose tolerance, and some cases accompanied with or without ketonuria. Histopathologically, eight cattle were diagnosed as chronic IDDM, while others were acute IDDM. The most characteristic lesions of the pancreas in chronic IDDM showed a decrease in the size and number of pancreatic islets, interlobular and interacinar fibrosis, mild lymphocytic insulitis, and vacuolation of a few islets. Almost all cells in the atrophied islets had a small amount of ungranulated cytoplasm. Immunohistochemical examination revealed that the atrophied islet cells did not react to anti-insulin antibody, but occasionally reacted to anti-glucagon or somatostatin antibodies. A few solitary islets with mild lymphocytic infiltration, necrotic islets with occasional calcification, and atrophied islets with mild fibrosis were also observed. A few islets consisted of many islet cells with vacuolated cytoplasm including a small number of insulin-positive granules. Accumulation of glycogen granules was occasionally observed in these islets. Islet fibrosis was due to the proliferation of collagen fibers reactive to both anti-collagen type I and type III antibodies. In acute IDDM, the major islets consisted of the cells with vacuolated cytoplasm indicating the degranulation of islet cells. These islets contained many islet cells with shrunken cytoplasm and karyorrhectic nuclei. Lymphocytic infiltration was frequently observed in the islets which consisted of many islet cells having karyorrhectic nuclei and vacuolated and severely degranulated cytoplasm. Immunohistochemically, islet cells with vacuolated cytoplasm had a small amount of insulin-positive granules, suggesting severe degranulation of beta-cells. An increase in acinar islet-cells and proliferation of ductal epithelial cells showing insulin-immunoreactivity were observed. Bovine IgG-immunoreactive islet cells were frequently seen in the vacuolated islets. In summary, pathological observations suggested that beta-cells were being destroyed by an inflammatory process which selectively affected the pancreatic islets. Lymphocytic insulitis and anti-bovine immunoreactive islet cells were thought to be the most significant changes in determining the etiology and pathogenesis of bovine IDDM, and suggested their role in anti-islet autoimmunity in this form of diabetes.

cDNA generated from lipopolysaccharide-stimulated bovine peripheral blood mononuclear cells was used to amplify and clone the bovine interleukin-1 receptor antagonist (IL-1ra) using primers derived from semi-conserved regions between human and mouse IL-1ra sequences. 5' and 3' terminal sequences of bovine IL-1ra were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of bovine IL-1ra demonstrated 80%, 78%, 78%, 77% and 76% homology with human, mouse, rat, rabbit and equine sequences, respectively. Recombinant bovine IL-1ra produced in Escherichia coli suppressed the growth inhibitory activity of bovine IL-1beta on A375 cells in a dose-dependent manner, indicating that the present bovine IL-1ra cDNA encodes biologically active proteins.

TNF-alpha has been reported to be induced in mice infected with Sendai virus. We evaluated the role of TNF-alpha in the virus infection. TNF-alpha was induced locally in proportion to virus titers in the lung. The activity was correlated with suppression of body weight gain. Passive immunization against TNF-alpha improved body weight gain and ameliorated pneumonic lesions in infected mice, and prevented them from lethal infection, but lung virus induced emaciation, pneumonic lesions and death were mediated by TNF-alpha.

This investigation was performed to determine whether primary cultures of mammary cells from lactating cows would sustain production of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF), and express mRNA for cytokines interleukin (IL)-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, interferon (INF)-tau, TNF-alpha, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by the reverse transcription-polymerase chain reaction (RT-PCR) in vitro. Cryopreserved mammary epithelial cells collected from cows at 1 week post calving were plated in collagen-coated 24-well culture plates (250,000 cells/well). IL-1 and IL-6 productions were measured using a A375 cell growth inhibition assay and a 7TD1 hybridoma proliferation assay, respectively. Production of IL-1 was demonstrated in mammary epithelial cells cultured with unsupplemented medium, but was not produced by cells cultured in medium supplemented with fetal bovine serum. IL-6 production in the conditioned medium was continued at steady level until day 14, whereas IL-6-like bioactivity was not detected in medium alone. TNF-like activity was not detectable in any experiments. This study also demonstrated the expression of mRNA for multiple cytokines including IL-1alpha, IL-1beta, IL-6, IL-10, TNF-alpha, and GM-CSF by RT-PCR in mammary cell cultures. The results indicate that bovine mammary epithelial cells of lactating cows produce IL-1 and IL-6 and have gene expression for multiple cytokines. This in vitro model will be useful to investigate the function and regulation of IL-1 and IL-6 in the lactating mammary gland.

As part of ongoing research into the conservation of wild avian species in Niigata Prefecture, helminthological examination was conducted between March and November 2002 in Niigata Prefectural Bird Protection Center, Japan. A total of 50 individuals of 28 avian species was investigated, with parasitic helminthes collected from 26 individuals. The parasitic helminths belonged to 13 nematode, two acanthocephalan, and three trematode genera, and unidenfied cestodes were also collected. Three genera, Epomidostomum (host: Anser albifrons), Viktorocara (host: Fulmarus glacialis) and Diomedenema (host: Ardea cinerea) were new locality records in Japan.

This study examined the levels of serum tumor necrosis factor (TNF), interleukin-1 (IL-1) and plasma cortisol levels in adult cows after intravenous administration of lipopolysaccharide (LPS) extracted from Escherichia coli. The adult cows exhibited clinical signs of endotoxic shock. The mean concentrations of serum TNF was 557.9 U/ml and serum IL-1 was 90.6 U/ml at 2 hr after LPS administration following severe endotoxic shock. Plasma cortisol concentrations increased immediately and a peak level was 131.4 ng/m l at 1 hr after LPS challenge. These results suggested that circulating concentrations of TNF, IL-1, and cortisol may be associated with clinical signs of endotoxemia in adult cows.

An intestinal carcinoid with multiple metastases was identified in a 5-year-old male Shih Tzu with a clinical history of anemia, fatigue, anorexia, vomiting, intermittent diarrhea, intestinal bleeding, and progressive emaciation. There was a yellowish-white mass 15 mm in diameter in the anterior jejunum and white nodules consistent with metastases in many organs. Histopathologically, the mass consisted of neoplastic cells arranged in lobules, trabeculae, or closely interdigitating islands of cells. Neoplastic cells were generally polygonal with round hyperchromatic nuclei, modest amounts of eosinophilic cytoplasm, and eosinophilic cytoplasmic granules. Mitoses were common. Rosette formations of tumor cells were apparent in metastatic tumors. Immunohistochemically, tumor cells stained positive for cytokeratin 13, synaptophysin, protein gene product 9.5, neuron-specific enolase, chromogranin A, calcitonin gene-related peptide, serotonin (5-HT), and Leu-7. Serum 5-HT concentrations for this dog were increased 10-fold compared with those of normal dogs. All findings were consistent with a diagnosis of a malignant intestinal carcinoid.

The effects of chlorpromazine (CPZ), pentoxifylline (PTX) and dexamethasone (DEX) on mRNA expression of lipopolysaccharide (LPS)-induced proinflammatory cytokines were examined in bovine peripheral blood mononuclear cells (PBMCs) in vitro. The expression of inflammatory cytokine mRNAs was analyzed by RT-PCR and Southern blot hybridization in bovine PBMCs. CPZ and DEX decreased the expression of cytokine mRNA (such as interleukin-1 beta and tumor necrosis factor-alpha) after stimulation with LPS in a dose-dependent manner. However, pretreatment with PTX had no inhibitory effect on the mRNA expression of proinflammatory cytokines. These results indicated that pretreatment with CPZ and DEX might be effective to reduce the production of LPS-induced inflammatory cytokines in bovine PBMCs in vitro.

A malignant pheochromocytoma with multiple metastases was diagnosed in a 7-year-old male wolfdog that resulted from a cross between an eastern timber wolf (Canis lupus lycaon) and an Alaskan malamute. A yellowish white neoplastic mass approximately 10 cm diameter was found in the right adrenal gland. The neoplasm penetrated through the wall of the caudal vena cava. A diagnosis of pheochromocytoma was established by histopathologic and immunohistochemical procedures. Immunohistochemically, the neoplastic cells expressed chromogranin A, substance P, synaptophysin, Leu-7, protein gene product 9.5, methionine- enkephalin, S100 protein, and galanin. Multiple metastatic tumors were found in the kidneys, spleen, lungs, heart, and liver.