Detection of bovine herpesvirus 4 in CD11b+ leukocytes of experimentally infected rabbits

The presence and numbers of bovine herpesvirus 4 (BoHV-4) infected CD11b+ leukocytes were investigated during experimental infections of New Zealand White rabbits by Fluorescence Activated Cell Sorter (FACS) analysis. Peripheral blood leukocytes (PBL) were collected every second day, and the cells were stained with phycoerythrin-labelled CD11b-specific mouse monoclonal antibody and fluorescein-conjugated bovine herpesvirus 4-specific mouse monoclonal antibody. The numbers of double-stained cells from PBLs of the control and inoculated groups were measured and compared in FACSTREK analyser. Double-stained cells were detected in the virus-inoculated group on postinoculation days (PID) 2-5 and 9-12. The results indicated that CD11b+ PBLs were permissive for BoHV-4 infection, and are probably the main reservoir of the virus during the latent period. The data did not indicate production of infectious viral particles, but virus-specific proteins were expressed on the surface of CD11b+ cells. The two waves of double-stained cells gave similar results to the PCR assays from serum samples, which showed the presence of viral DNA in the serum on the same days when virus-infected CD11b cells were also present. Productive BoHV-4 infection of mast cells or undifferentiated leukocytes in the bone marrow and the antiviral immune response might be responsible for this periodic appearance of the virus in CD11b+ PBLs and in the serum. The paper provides evidence that CD11b+ PBLs are the main target cell populations in the blood for BoHV-4.     

Fluctuation in the concentration of sex steroids and aflatoxin B1 in the seminal plasma of boars and its relation to sperm production

The concentration of testosterone, 17-beta oestradiol and aflatoxin B1 were studied in the semen plasma of 21 boars of four breeds for the period of twelve months. The following spermiological parameters were investigated: semen volume, sperm concentration, percentage of abnormal spermatozoa, and survival of spermatozoa. The fertilizing capacity of ejaculates was evaluated according to the conception rate of sows and gilts after the first insemination, according to the average number of piglets per litter and average number of live-born piglets per litter. The highest aflatoxin B1 residues in sperm were recorded in March to May and were related with aflatoxin concentration in feed ration. The group of boars with fertility disorders had more aflatoxin in their sperm (up to 100 pmol . l-1), lower sperm concentration, lower survival of spermatozoa, and a larger proportion of abnormal spermatozoa. The year season had a significant influence on the concentration of the hormones. The highest average value of testosterone (10.2 +/- 1.28 nmol) was obtained in autumn and lower values were recorded in winter. The changes in 17-beta estradiol concentration were similar to the changes in testosterone content, with the maximum value in November (0.249 nmol X 1(-1]. The boars with reproduction disorders had a significantly lower concentration of 17-beta oestradiol. Significant correlations were found between the concentration of the hormones, semen volume, and sperm concentration. 17-beta oestradiol also had a significant positive correlation to abnormal spermatozoa and to the activity of aspartate aminotransferase.     

Acute viral hepatitis in California sea lions

Acute viral hepatitis was diagnosed in five California sea lions (Zalophus californianus) stranded along the Los Angeles coast. Light microscopy revealed large nuclear inclusion bodies in hepatocytes. Electron microscopy provided evidence that these inclusion bodies were composed of adenovirus-like virions. Attempts to grow the virus in cell culture systems were unsuccessful.     

Comparative studies on the distribution and population of immunocompetent cells in bovine hemal node, lymph node and spleen

The distribution and population of immunocompetent cells in bovine hemal node, mesenteric lymph node and spleen were analyzed comparatively by immunohistochemistry and flow cytometry. Many CD8(+) cells, CD172a(+) cells and γδ T cells were found in the lymphatic cord along the sinus of the hemal node and the splenic red pulp. A few CD8(+) cells and γδ T cells were distributed diffusely in the paracortex and medullary cord of the mesenteric lymph node. Many germinal centers were recognized in the lymphatic regions such as the cortex and white pulp of these lymphoid organs. The populations of CD8(+) cells and γδ T cells in the hemal node and the spleen were higher than those of the mesenteric lymph node. In addition, the populations of CD21(+) cells and MHC class II(+) cells in the hemal node and the mesenteric lymph node were higher than those of the spleen. The results suggest that the hemal node has an important role in both cellular and humoral immunity as well as the lymph node and the spleen in cattle.     

In vivo and ex vivo assessment of the interaction between ivermectin and danofloxacin in sheep

The impact of an efflux pump-related interaction between ivermectin and danofloxacin on their intestinal transport (ex vivo) and disposition kinetics (in vivo) was assessed. Eighteen male Corriedale sheep were randomly assigned to one of three groups. Animals in Group A received 0.2mg/kg ivermectin by SC injection, those in Group B were given 6 mg/kg danofloxacin SC on two occasions 48 h apart and those in Group C were treated with both compounds at the same rates. Plasma concentrations of ivermectin and danofloxacin were measured by HPLC using fluorescence detection. Ex vivo intestinal drug transport activity was measured by the use of the Ussing chamber technique. Plasma concentrations of ivermectin in the first 6 days after injection tended to be higher in Group C than Group A. Contemporaneous treatment with ivermectin significantly increased systemic exposure to danofloxacin (AUC values were 32-35% higher) and prolonged the elimination half-life of danofloxacin (40-52% longer). Ex vivo, incubation with ivermectin significantly decreased the efflux transport of rhodamine 123, a P-glycoprotein substrate, in sheep intestine, but no significant effect of danofloxacin on transport activity was observed. Evaluation of the interaction of danofloxacin with the breast cancer resistance protein (BCRP) showed that pantoprazole and ivermectin significantly decreased danofloxacin secretion in the rat intestine. Thus, the ivermectin-induced reduction of danofloxacin efflux transport observed in this study may involve BCRP activity but the involvement of P-glycoprotein cannot be ruled out.     

Analytical and Clinical Validation of a Time-resolved Immunofluorometric Assay (TR-IFMA) for Canine C-reactive Protein in Serum

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of C-reactive protein (CRP) in canine serum. CRP was isolated from canine acute-phase serum by affinity chromatography on agarose coupled with phosphorylethanolamine. This isolated dog CRP was used as standard to calibrate the assay. Intra-assay and inter-assay coefficients of variation were in the ranges 5.3–7.1% and 4.8–13.3%, respectively. Accuracy, evaluated by adding 2 and 10 μg/ml of CRP to serum samples, provided recoveries of 99.9% and 106.8%. High correlation was found between CRP measurements by TR-IFMA and a by commercial enzyme-linked immunosorbent assay (R² = 0.98). The limit of detection for the TR-IFMA method was 0.000067 μg/ml and the measurement of CRP in serial dilutions of acute-phase dog sera generated curves with the same slope as the one constructed with purified CRP. The TR-IFMA provides a precise, accurate and highly sensitive assay for CRP determination in dog samples. CRP levels in dogs with different diseases ranged between 10.2 and 210.7 μg/ml and were significantly higher than those observed in healthy dogs (< 7.1 μmg/ml).     

Comparative fluorescein angiography of the normal sheep and goat ocular fundi

Fluorescein angiography without sedative or anesthetic agents was evaluated in 20 normal goats and 20 normal sheep. All of the angiographic phases were observed using 20 mg/kg fluorescein IV in both species. Fundus fluorescein angiography results revealed wide stars of Winslow in the tapetal fundus, central or marginal flow during the first part of the arterial phase, delayed filling of the focal areas in the choroid near the optic disc that often coincided with others in the disc, and lack of evidence of the 'striate area' in the tapetal fundi. In goats, the angiographic times were 6.54+/-1.25 s for the arterial phase (TA), 7.80+/-1.37 s for the arterio-venous phase (TAV), and 14.13+/-2.01 s for the venous phase (TV). I1: 1.30+/-0.30 s (time elapsing between TA and TAV), and I2: 6.20+/-1.60 s (time elapsing between TAV and TV). In sheep, times were 9.54+/-2.18 s TA, 11.73+/-2.10 s TAV, and 20.86+/-2.74 s TV. I1: 2.04+/-0.75 s and I2: 8.98+/-2.47 s, respectively. Due to the large size of the fundic vessels in sheep and goats, fluorescein angiography of the retinal vasculature can facilitate the study of the different vascular diseases in these species.     

Structural changes in femoral bone tissue of rats after subchronic peroral exposure to selenium

Background

The role of selenium (Se) on bone microarchitecture is still poorly understood. The present study aims to investigate the macroscopic and microscopic structures of femoral bone tissue in adult male rats after subchronic peroral administration of Se.

Methods

Twenty one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group (Se group) young males were exposed to 5 mg Na₂SeO₃/L in drinking water, for 90 days. Ten one-month-old males without Se administration served as a control group. At the end of the experiment, macroscopic and microscopic structures of the femurs were analysed using analytical scales, sliding instrument, and polarized light microscopy.

Results

The body weight, femoral length and cortical bone thickness were significantly decreased in Se group rats. These rats also displayed different microstructure in the middle part of the femur, both in medial and lateral views, where vascular canals expanded into the central area of the bone while, in control rats, these canals occurred only near the endosteal surfaces. Additionally, a smaller number of primary and secondary osteons was identified in Se group rats. Histomorphometric analyses revealed significant increases for area, perimeter, maximum and minimum diameters of primary osteons’ vascular canals but significant reductions for all measured variables of Haversian canals and secondary osteons.

Conclusions

Se negatively affected the macroscopic and microscopic structures of femoral bone tissue in adult male rats. The results contribute to the knowledge on damaging impact of Se on bone.     

Effects of Garlic and Ginger Oils on Hematological and Biochemical Variables of Sea Bass Dicentrarchus labrax

Abstract

This study was conducted to investigate the effects of garlic and ginger oils on hematological and biochemical health characteristics of sea bass Dicentrarchus labrax. Fish were exposed to garlic oil (0.01 or 0.02 mL/L), ginger oil (0.01 or 0.02 mL/L), or a combination of the two oils (each oil at a concentration of 0.005 or 0.01 mL/L) for 96 h via bath immersion. Results showed that the red blood cell count, hematocrit (%), hemoglobin (Hb) concentration (g/dL), mean corpuscular volume (μm³), mean corpuscular Hb (pg), and mean corpuscular Hb concentration (%) were not significantly affected by herb oil exposure. However, some changes in biochemical variables were observed. Sea bass exposed to the 0.005-mL/L garlic oil–ginger oil mixture exhibited a significant increase in serum glucose. Serum total protein and albumin levels decreased in sea bass that were exposed to a garlic oil–ginger oil mixture (0.005 or 0.01 mL/L) or to garlic oil at 0.02 mL/L. Serum globulin levels decreased and triglyceride levels increased in sea bass exposed to 0.02-mL/L garlic oil or to the 0.01-mL/L mixture. The serum lipase level decreased and the cholesterol level increased in fish that were exposed to 0.02-mL/L garlic oil. In summary, ginger oil at 0.01–0.02 mL/L can be used without negative effects, while the garlic oil or garlic oil–ginger oil mixture should be applied at a concentration below 0.005 mL/L for bath immersion of sea bass. This is the first study to examine how garlic oil and ginger oil exposure via bath immersion affects the hematological and biochemical status of sea bass.

Received March 19, 2012; accepted July 2, 2012