Pathological changes of tracheal mucosa in chickens infected with lentogenic Newcastle disease virus

Forty-day-old specific-pathogen-free chickens were exposed by aerosol to lentogenic Newcastle disease virus (NDV) and observed for 24 days for pathological changes in the tracheal mucociliary system. Specific fluorescence of NDV antigen was observed through day 5 postexposure (PE) in the cytoplasm of the tracheal epithelium and desquamated epithelium in the lumen. On day 1 PE, scanning electron microscopy revealed hypertrophy of goblet cells and small patches of the deciliated epithelium scattered mainly around the openings of mucous glands. The deciliated area of tracheal surface increased through day 4 PE. Light microscopy showed small vacuoles containing lymphocytes and heterophils in the epithelial layer. Immature epithelium proliferated in some areas. On days 5 and 6 PE, ciliated areas of the trachea tended to increase as a result of regeneration of the epithelium, still leaving many nonciliated patches of various sizes. On and after day 8 PE, there remained plaques with nonciliated flat epithelium, but most areas were covered with well-ciliated epithelium. Non-ciliated plaques were observed until day 24 PE, but they gradually decreased in size. These plaques were covered by a single layer of flat epithelium and were formed upon lymph follicles in subepithelial tissue.     

Separating component parts of soil respiration under <Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> plantation in the Taihang Mountains,China

Partitioning the respiratory components of soil surface CO₂ efflux is important in understanding carbon turnover and in identifying the soil carbon sink/source function in response to land-use change. The sensitivities of soil respiration components on changing climate patterns are currently not fully understood. We used trench and isotopic methods to separate total soil respiration into autotrophic (R _A ) and heterotrophic components (R _H ). This study was undertaken on a Robinia pseudoacacia L. plantation in the southern Taihang Mountains, China. The fractionation of soil ¹³CO₂ was analyzed by comparing the δ¹³C of soil CO₂ extracted from buried steel tubes with results from Gas Vapor Probe Kits at a depth of 50 cm at the preliminary test (2.03‰). The results showed that the contribution of autotrophic respiration (fR _A ) increased with increasing soil depth. The contribution of heterotrophic respiration (fR _H ) declined with increasing soil depth. The contribution of autotrophic respiration was similar whether estimated by the trench method (fR _A , 23.50%) or by the isotopic method in which a difference in value of ¹³C between soil and plant prevailed in the natural state (RC, 21.03%). The experimental error produced by the trench method was insignificant as compared with that produced by the isotopic method, providing a technical basis for further investigations.     

Thyroid Hormone-disrupting Effects and the Amphibian Metamorphosis Assay

There are continued concerns about endocrine-disrupting chemical effects, and appropriate vertebrate models for assessment of risk are a high priority. Frog tadpoles are very sensitive to environmental substances because of their habitat and the complex processes of metamorphosis regulated by the endocrine system, mainly thyroid hormones. During metamorphosis, marked alteration in hormonal factors occurs, as well as dramatic structural and functional changes in larval tissues. There are a variety of mechanisms determining thyroid hormone balance or disruption directly or indirectly. Direct-acting agents can cause changes in thyroxine synthesis and/or secretion in thyroid through effects on peroxidases, thyroidal iodide uptake, deiodinase, and proteolysis. At the same time, indirect action may result from biochemical processes such as sulfation, deiodination and glucuronidation. Because their potential to disrupt thyroid hormones has been identified as an important consideration for the regulation of chemicals, the OECD and the EPA have each established guidelines that make use of larval African clawed frogs (Xenopus laevis) and frog metamorphosis for screening and testing of potential endocrine disrupters. The guidelines are based on evaluation of alteration in the hypothalamic-pituitary-thyroid axis. One of the primary endpoints is thyroid gland histopathology. Others are mortality, developmental stage, hind limb length, snout-vent length and wet body weight. Regarding histopathological features, the guidelines include core criteria and additional qualitative parameters along with grading. Taking into account the difficulties in evaluating amphibian thyroid glands, which change continuously throughout metamorphosis, histopathological examination has been shown to be a very sensitive approach.     

Effect of FMD vaccination on semen quality parameters in Karan Fries and Murrah buffalo bulls

Effect of Foot and Mouth disease (FMD) vaccination was studied on semen quality parameters of 19 Karan Fries (KF) and eight Murrah (MU) breeding bulls during the period 2002 to 2004 at Artificial Breeding Complex, NDRI, Karnal. A total of non-vaccinated 155 KF and 72 MU bulls' ejaculates were taken as control, while 169 KF and 51 MU bulls' ejaculates, collected after vaccination, were used to study the effect of vaccination stress. The results showed that FMD vaccination had no significant (P > 0.05) effect on ejaculate volume and total volume per day of semen in both KF and MU bulls. Volume of semen increased slightly during post-vaccination period in both the breeds. After FMD vaccination, there was significant (P < 0.01) decrease in mass activity (2.27 ± 0.06 vs. 1.67 ± 0.07 and 2.49 ± 0.09. vs. 1.75 ± 0.10, for KF and MU, respectively), initial motility (56.89 ± 0.03% vs. 44.62 ± 0.02% and 62.26 ± 0.04% vs. 47.08 ± 0.05%, for KF and MU, respectively), sperm concentration (754.19 ± 23.96 vs. 554.14 ± 22.95 × 10⁶/ml and 848.61 ± 33.65 vs. 571.57 ± 39.99 × 10⁶/ml, for KF and MU, respectively), and total sperm output per ejaculate (3,685.94 ± 158.40 vs. 2,781.54 ± 151.70 × 10⁶ and 2,218.75 ± 133.14 vs. 1,582.84 ± 158.20 × 10⁶, for KF and MU, respectively). Application of FMD vaccine had significantly (P < 0.05) adverse effect on most of the seminal attributes during post-vaccination in KF and MU buffalo bulls. So, the spermiograms affected following vaccination suggest that in bovines, the semen collection and preservation should be suspended till normal fertility of sperm is restored to avoid the failure of conception from artificial insemination using such semen.     

Plasma pharmacokinetics and milk levels of ceftriaxone following single intravenous administration in healthy and endometritic cows

Pharmacokinetics and milk levels of ceftriaxone were studied in healthy and endometritic cows following single intravenous administration. The drug was detected up to 8 h of dosing in plasma of healthy and endometritic cows and the drug disposition followed three-compartment open model. The values of Vd_area, AUC, t_1/2β, Cl_B, MRT and P/C ratio were 0.50 ± 0.19 L.kg⁻¹, 62.2 ± 23.3 μg.ml⁻¹.h, 1.02 ± 0.07 h, 0.30 ± 0.09 L.kg⁻¹.h⁻¹, 1.55 ± 0.25 h and 0.52 ± 0.27, respectively, in healthy and 1.55 ± 0.52 L.kg⁻¹, 37.0 ± 17.1 μg.ml⁻¹.h, 1.56 ± 0.25 h, 0.56 ± 0.14 L.kg⁻¹.h⁻¹, 2.14 ± 0.34 h and 1.44 ± 0.60, respectively, in endometritic cows. The drug was detected in milk for 36 h after administration. For MIC₉₀ of 0.5 μg.ml⁻¹ the most appropriate dosage for ceftriaxone, would be 9.0 mg.kg⁻¹ repeated at 6 h intervals for the treatment of endometritis in cows.     

Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen <Emphasis Type="Italic">Dichelobacter nodosus</Emphasis>

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.     

Surveillance and characterization of Newcastle disease viruses isolated from northern pintail (Anas acuta) in Japan during 2006-09

A total of 38 Newcastle disease virus (NDV) isolates were obtained from 6060 fecal samples from northern pintail (Anas acuta) ducks collected in the Tohoku district in Japan during 2006-09. One isolate from each sampling location and date was selected for a total of 38 isolates, then 15 of these were characterized for their pathogenicity by mean death time of minimum lethal dose (MDT/MLD) using chicken embryos and by plaque formation on chicken embryo fibroblasts. Furthermore, nine isolates were randomly selected from these 15 isolates, and the fusion protein genes were sequenced to characterize amino acid sequences around the cleavage site. All 15 were confirmed to be nonvirulent by MDT/MLD test, and nine isolates were also confirmed as nonvirulent by the cleavage site of the fusion protein 112G/E-K/R-Q-G/E-R*L117 that was specific for nonvirulent NDVs. The characteristics of nine isolates identified by phylogenic analysis of the fusion protein gene indicated that the isolates belong to genotype I or II. In addition, we also isolated 68 avian influenza viruses and 28 other hemagglutinating viruses. Our data indicate that northern pintails are subclinically infected by, perpetuate, and distribute NDV along with different subtypes of avian influenza viruses and other hemagglutinating viruses during their migrations across vast areas over the Northern Hemisphere to Japan.     

Cynomolgus macaque CYP4 isoforms are functional, metabolizing arachidonic acid

Cytochrome P450 (CYP) is important for metabolism of not only xenobiotics such as drugs, but also endogenous compounds including arachidonic acids. CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 have been identified in cynomolgus macaque, an animal species widely used for investigation of drug metabolism due to its evolutionary closeness to human. However, their metabolic functions have not been investigated. In this study, proteins were heterologously expressed in Escherichia coli and characterized by metabolic assays using arachidonic acids as substrates that are metabolized by CYP4 isoforms in human. The results showed that all four CYPs metabolized arachidonic acids. Therefore, cynomolgus macaque CYP4A11, CYP4F3v2, CYP4F11, and CYP4F45 are functional enzymes.     

Circulating oxidative stress status in dromedary camels infested with sarcoptic mange

Oxidative stress is an imbalance between radical-generating and radical-scavenging activity, resulting in oxidation products and tissue damage. This study was aimed to evaluate the status of oxidative stress indices in blood of camels naturally infested with S. scabiei. Forty-seven male camels (Camelus dromedaries) were divided according to the extent of the infested area with Sarcoptes scabiei into four groups, mild (MID, n = 12), moderate (MOD, n = 10), severely infested (SEV, n = 10) and healthy control group (n = 15). Blood was used for determination of red cell count (RBC), hemoglobin (Hb), packed cell volume (PCV), serum nitric oxide (NO^•, a free radical), ascorbate and albumin concentrations, and erythrocytic values of malondialdehyde (MDA, a marker of lipid peroxidation), protein carbonyls (PC, an indicator of protein oxidation), glutathione (GSH) superoxide dismutase (SOD) and catalase (CAT). Decreased levels (P < 0.05) of RBC, Hb, PCV, albumin and ascorbate were noticed in MOD and SEV compared to controls with the lowest values (P < 0.05) in SEV except for ascorbate, where MOD did not differ from SEV. Compared to controls, NO^• gradually increased (P < 0.05) in MID followed by MOD and SEV, whereas MDA and PC were higher (P < 0.05) in MOD and SEV. PC was higher (P < 0.05) in MOD than SEV. In addition, the antioxidants GSH, SOD and CAT were higher (P < 0.05) in MID and lower (P < 0.05) in MOD and SEV compared to controls. GSH was lower (P < 0.05) in SEV compared to MOD. Besides, Hb was negatively correlated with NO^• (r = −0.68, P < 0.001), MDA (r = −0.53, P < 0.001) and PC (r = −0.73, P < 0.001). In conclusion, dromedary sarcoptosis is accompanied by a state of oxidative stress process, which increased by increasing the area of infestation, and may contribute to the pathogenesis of the disease.