To investigate the clinical utility of C-reactive protein (CRP) determination in dogs, its plasma concentration was measured by a laser nephelometric method in 928 dogs with various diseases, and was compared with other inflammatory parameters. CRP concentration was elevated in various inflammatory diseases, this was most frequently observed in cases with neoplastic and immune-mediated diseases. All cases of pyometra, panniculitis, acute pancreatitis, polyarthritis, and hemangiosarcoma showed significantly elevated CRP concentrations. On the other hand, the CRP concentration was elevated only in few cases of neurological diseases such as epilepsy, meningoencephalitis, and hydrocephalus and endocrine diseases such as hypothyroidism, hyperadrenocorticism, and diabetes mellitus. Only a weak correlation was observed between the CRP concentration and white blood cell (WBC) counts (r=0.44) but no correlation with band neutrophil counts. There was no correlation between the CRP and albumin concentrations, but a weak negative correlation (r=-0.40) when excluding chronic intestinal diseases and nephrotic syndrome, which can cause protein loss. Thus, CRP can be useful to detect inflammations that cannot be detected by WBC and, or band neutrophil counts, suggesting that the examination of CRP concentration is essential as routine diagnostic test.
Characteristics of the signal transduction in Platelet activating factor (PAF)-activated platelets and effects of anti-platelet agents on this response were investigated in vitro for potential therapeutic applications in canine endotoxemia. Blockade of the PAF receptor by a specific blocker has the strongest inhibitive effect on the PAF-induced platelet reactions. The response was also inhibited by either Ca2+ channel blockers or prostaglandin E1.
In the present study, we determined the levels of cytokines produced by camel (Camelus bactrianus) peripheral blood mononuclear cells (PBMCs) in response to live attenuated Brucella abortus (B. abortus) S19 vaccine. Seven camels were vaccinated with commercial B. abortus S19 vaccine, and their cytokine responses were determined using a real-time PCR assay. Cytokine responses to B. abortus S19 were examined at 6 hr, 48 hr and 1, 2 and 3 weeks post-vaccination. Serological tests were performed to further confirm these immune responses. The results revealed that IFN-γ and IL-6 were upregulated during the first week post-vaccination. Low level expressions of IL-1α, IL-1β, TNFα and IL-10 and no expression of IL-2 and IL-4 were observed compared with the control camels. The findings showed that B. abortus stimulates cell-mediated immunity by directly activating camel Th1 cells to secrete IFN-γ. This quantification of cytokine expression in camels is essential for understanding of Camelidae disease development and protective immune responses. This is the first report of in vivo camel cytokine quantification after vaccination.
The aim of this study was to clarify the hormonal and immunological changes in periparturient dairy cows with excessive weight loss following calving. We analyzed leukocyte populations and the peripheral blood mononuclear cell (PBMC) mRNA of IFN-γ, IL-4, growth hormone (GH)-receptor (R) and insulin -R and measured the GH and insulin concentrations. Ten dairy cows fed in one herd were used in this study and were divided into two groups. Five cows that experienced loss of body weight following calving of over 10% comprised the Weight Loss Group, and the remaining five cows, which experienced loss of body weight of 5% or less following calving, served as the Control Group. The Weight Loss Group had high serum cholesterol and low blood urea nitrogen levels compared with the Control Group throughout the period of observation. In regard to the leukocyte populations, there were significantly fewer CD4+CD45R-T cells in the Weight Loss Group 1 to 2 months after calving compared with the Control Group. The percentage of IFN-γ/IL-4 among PBMCs in the Weight Loss Group tended to be lower than in the Control Group throughout the observation period. In the Weight Loss Group, the levels of GH-R during month 2 were lower than in the Control Group. We concluded that excessive loss of body weight in dairy cows following calving is associated with depressed cellular immune function.
The purpose of the present study was to develop a technique for biopsy of the kidneys of cattle and to evaluate its usefulness and safety based on 25 consecutive biopsies. Using a handheld 14G spinal biopsy needle, the right kidney was biopsied under ultrasound control by one operator. To assess the immediate effects of renal biopsy, twenty cattle were necropsied shortly after the procedure and examined. The remaining five animals were biopsied and examined daily for nine days and then necropsied. One pass was made for biopsy of the right kidney. No immediate or delayed macroscopic hematuria was observed post-biopsy. Six animals had a thin subcapsular hematoma (less than 2 cm in diameter). We conclude that percutaneous ultrasound-guided biopsy of the right kidney is safe, fast, cost-effective and practical as long as it is performed properly.
Plasma metabolites and peripheral lymphocyte subsets were measured in ten diabetic and ten control dogs to investigate their significances as indicators to evaluate immune states in the diabetic dogs. Diabetic dogs were treated with insulin injections, however their plasma glucose and fructosamine concentrations were significantly higher than those of the controls. There were no significant differences in counts of total white blood cells (WBC) and lymphocyte CD8+ cells (cytotoxic T cells) between the control and the diabetic dogs. In the diabetic dogs, the counts of CD3+ (T cells), CD4+ (Helper T cells) and CD21+ (B cells) cells and the peripheral lymphocytes CD4/CD8 ratio were significantly lower than those in the control dogs. We confirmed abnormality of lymphocyte subsets in insulin treated diabetic dogs and it may relate to depression of immunocompetence and high susceptibility to common infectious diseases.
The effects of mycotoxins on mitogen-stimulated proliferation of bovine peripheral blood mononuclear cells (PBMCs) were investigated. Aflatoxin B1 (AFB1), deoxynivalenol (DON) and zearalenone (ZEN) were added to cultures of PBMCs, and the proliferation responses were measured using MTT bioassays. Suppression of the proliferation of calf PBMCs by AFB1 and DON was significantly stronger than that of cow PBMCs, whereas there were no differences in suppressive effects on PBMCs from Holstein and Japanese Black calves and cows. The suppressive effect was greatest in the order of DON, AFB1 and ZEN, and the effects of DON and AFB1 seemed to be dose-dependent. The results suggest that some mycotoxins directly suppress proliferation of bovine PBMCs.
Spontaneously Diabetic Torii (SDT) rats were discovered from SD rats and represent a confirmed spontaneous type 2 diabetes mellitus model. We investigated the effect of repeated stress in early childhood on SDT rats fed a high-fat diet, on locomotor activity and on the onset of diabetes mellitus. Regarding stress, a water immersion-restraint stress (WIRS) burden was applied 10 times every other day from 4 weeks of age. The results of the study showed, that the locomotor activity of the young SDT rats was clearly lower than that of the SD rats, and their locomotor activity was inferred to be congenitally low. In addition, the stress-burdened SDT rats showed delayed onset of diabetes mellitus and impaired glucose tolerance compared with the rats not receiving stress burden. The locomotor activity of SDT rats is less than that of SD rats, and they SDT rats are thought to have poor at spontaneous energy expenditure. On the other hand, the feeding efficiency of the WIRS-burdened SDT rats was reduced, and in comparison with the SDT rats with no WIRS burden, energy expenditure was increased; this is suggested to influence the onset of diabetes mellitus.
An adult male white eared-pheasant (Crossoptilon crossoptilon) at a Japanese zoo exhibited lethargy and emaciation. Microscopic examination of a blood smear revealed a haemosporidian parasitic infection. Based on the morphological characteristics and molecular analysis of the parasite, it was identified as Plasmodium (Bennettinia) juxtanucleare. This is the first report of P. juxtanucleare infection in bird species belonging to the genus Crossoptilon. Caution against avian malaria infection is required for the conservation of endangered bird species in zoos.
In the canine liver with portosystemic shunts (PSS), focal lesions consisting of cells with cytoplasmic brown pigments and lipid vacuoles are often observed in the hepatic parenchyma. Termed lipogranulomas, their histopathological characteristics have been little studied. In the present study, we examined liver biopsy samples from 144 dogs (age: 3 months-16 years; 65 PSS and 79 non-PSS cases), and investigated the histopathological characteristics, incidence, and density of lipogranulomas. Lipogranulomas were detected histopathologically in 55.4% of PSS dogs. The lesions were then grouped into 3 types according to the amount of cytoplasmic lipid vacuoles and brown pigments. The pigments were positive for Berlin blue, PAS, and Sudan black B, but negative with the Hall method. The majority of the cells were immunohistochemically positive for macrophage scavenger receptor, class A (MSR-A), while no cells were positive for hepatocyte-antigen and albumin. The cytoplasmic pigments were recognized as electron-dense microgranular materials by electron microscopy. The incidence of lipogranulomas was significantly higher in the PSS group than non-PSS group when dogs less than 1 year old were excluded. The lipogranuloma density in the liver was significantly higher in the PSS group. It is concluded that lipogranulomas are frequently observed in liver biopsies of canine PSS especially in dogs more than 1 year old. The lesions consisted of Kupffer cells and/or macrophages, and the cytoplasmic brown pigments are ceroid and hemosiderin. The pathogenesis of lipogranuloma in PSS needs to be clarified.
Device-associated infections are serious complications, and their prevention is an issue of considerable importance. Since biofilms are responsible for these refractory infections, effective methods to inhibit biofilm formation are required. In this investigation, stainless steel plates with and without 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, i.e., poly (MPC-co-n-butyl methacrylate) (PMB) coating, were incubated in a medium containing bacteria. In the course of incubation, half of the specimens received antibiotics. The specimens were stained for nucleic acid and polysaccharides, and then examined with a confocal laser scanning microscope. The numbers of bacteria on the specimen surfaces were evaluated by an ATP assay. On the surfaces of the specimens without PMB coating, the formation of a biofilm enveloping bacteria was confirmed. The addition of antibiotics did not effectively decrease the number of bacteria. On the other hand, on the surfaces of the specimens with PMB coating, no biofilm formation was observed, and the number of bacteria was significantly decreased. The addition of potent antibiotics further decreased the number of bacteria by 1/100 to 1/1000 times. The PMB coating combined with the validated use of antibiotics might provide a method for the simultaneous achievement of biocompatible surfaces of devices and the prevention of device-associated infections.
Manganese deficiency results in neurological and skeletal defects, together with ultrastructural disarrangement of the retina in rats. Wild birds show a range of Mn concentrations in their tissues, including the liver, raising the possibility of Mn-related disorders in the wild. Electroretinography (ERG) provides a useful noninvasive approach to evaluate visual function. This method is especially useful in birds, as objective analysis of them is very difficult, while they have well-developed vision. In this study, we carried out a convenient and reliable ERG recording using a contact lens electrode with a built-in light source (LED electrode) of Japanese quail (Coturnix coturnix japonica) fed a Mn-deficient diet. After 10 min light adaptation, single-flash and flicker cone responses were reproducibly recorded to cause an intensity-dependent increase in amplitude of both a-wave and b-wave in single-flash ERG. Mn-deficient feeding markedly decreased the Mn concentration in the liver by almost half in 3 to 6 weeks, followed by body weight loss in 13 to 15 weeks. Implicit time of a-wave and b-wave cone response by single-flash stimulation was significantly delayed in quail with a Mn depletion from 3 to 6 weeks. Every cone response of the Mn-deprived quail had a tendency to decrease amplitude. The ultrastructure of cone photoreceptor cells was disorganized by Mn deficiency, including changes in outer segment discs of photoreceptor cells. These results suggest the essential role of Mn in the integrity of the retinal function of birds.
The scrapie prion protein (PrP27-30) is a crucial component of the prion and is responsible for its transmissibility. Structural information on this protein is limited because it is insoluble and shows aggregated properties. In this study, PrP27-30 was effectively dispersed using sonication under the weak alkaline condition. Subsequently, the small PrP27-30 aggregates were subjected to different pH, heat, and denaturing conditions. The loss of proteinase K (PK) resistance of PrP27-30 and prion infectivity were monitored along with spectroscopic changes. Prion inactivation could not be achieved by the loss of PK resistance alone; a significant loss of the PrP27-30 amyloid structure, which was represented by a decrease in thioflavin T fluorescence, was required for the loss of transmissibility.
In this study, we evaluated whether five rapid antigen detection kits for human influenza could be used for the diagnosis of equine influenza (EI). Limiting dilution analyses showed that Directigen Flu A+B and ESPLINE INFLUENZA A&B-N had the highest sensitivities to equine-2 influenza viruses (EIVs) among the kits investigated. From the results of virus detection in nasal swabs taken from horses infected with EIV, these two kits could produce positive results in reasonable agreement with those obtained by virus isolation or RT-PCR, suggesting that these kits could be useful for rapid diagnosis of EI in the field. However, from the viewpoint of specificity for EIV, Espline seems to be superior to Directigen.