Rearing complexity affects branching of dendrites in the visual cortex of the rat

Higher-order dendritic branching is considerably greater in Golgistained neurons from the occipital cortex of rats reared in groups in a complex environment than in similar neurons of littermates reared individually in laboratory cages have intermediate amounts of branching, while lower-order branching did not appear to be affected by any rearing environment.     

New method using quantitative PCR to follow the tick blood meal and to assess the anti-feeding effect of topical acaricide against Rhipicephalus sanguineus on dogs

A 28-day study was conducted to assess the dynamic of blood feeding by Rhipicephalus sanguineus ticks on dogs treated or not with a novel topical combination of fipronil, amitraz and (S)-methoprene. Dogs were infested weekly through exposure to ticks in crates for 4 h. Ticks were then counted in the crates at 2 h and 4 h post dog exposure. Ticks were also counted and removed from the dogs at 2 h, 4 h, 6 h, 12 h and 24 h post tick exposure. The inhibition of blood feeding was assessed by both tick quantification and designing and performing a quantitative PCR (qPCR) to detect the canine hydroxymethylbilane synthase (HMBS) gene in ticks. The percentage of repellency sensu lato based on the ticks collected in crates at 2 h varied from 4.7% at day 28 to 48.3% at day 7. The immediate mortality rate of the ticks expelled at 2 h varied from 1.5% at day 21 to 31.7% at day 7. The efficacy calculation showed that the acaricidal combination started to kill ticks in as little as 2 h. The average efficacy reached 90.0% at 12 h post crate challenges and 100% at 24 h post exposure in crates. The inclusion of an internal amplification control was used to ensure that no significant template-derived PCR inhibition (≤6.2%) affected the overall results. The reduction of blood feeding was significant at 4 h (>80.0%) and >99.0% at 24 h post tick exposure in the crate. The high repellency rate and the lethal efficacy of CERTIFECT^® resulted in significantly fewer live attached ticks, consequently reducing blood intake and fluid exchanges.     

Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.     

Clinical evaluation of a peptide-ELISA based upon N-terminal B-cell epitope of the VapA protein for diagnosis of Rhodococcus equi pneumonia in foals

A total of 227 field samples from naturally exposed foals aged between 3 weeks and 6 months were used in an evaluation of a peptide-based enzyme-linked immunosorbent assay (ELISA) for diagnosis of Rhodococcus equi infection. A biotinylated peptide derived from the virulence-associated protein A (VapA) of R. equi, a horse pathogen, was synthesized and designated as PN11-14. The peptide corresponds to the N-terminal B-cell epitope TSLNLQKDEPNGRASDTAGQ of the VapA protein. Based upon a serum immunoglobulin (Ig)G titre of 512 as a positive cut-off value for the R. equi infection, the ELISA provided the overall sensitivity of 47.62%, specificity of 69.67% and an accuracy of 59.47% with a positive predictive value of 57.47% for true R. equi pneumonia. The assay was improved by detecting VapA-specific IgGb antibodies against N-terminal B-cell epitope of the VapA protein rather than IgG antibodies. The VapA-IgGb ELISA showed the overall sensitivity of 70.47%, specificity of 72.13% and accuracy of 71.36% with a positive predictive value of 68.52%. Diagnosis of R. equi disease in 6-week-old foals showed that the VapA-IgGb ELISA provided an increasing trend (P=0.0572) in sensitivity of 82.4% in comparison with the VapA-IgG ELISA which showed the sensitivity of 58.8%. However, differences in specificity of both tests were statistically insignificant (P=0.357) as analysed by the McNemar test. These results indicated that detection of VapA-specific IgGb antibodies may be a better predictor of R. equi disease in foals.     

Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus.

Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter-assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.     

Relationship between vitamin E content and susceptibility to lipid peroxidation in tissues of the newly hatched chick.

1. The effect of supplementing the diet of the parent hen with vitamin E on the vitamin E content of the yolk and of embryonic and neonatal tissues was evaluated and the effects of elevated tissue concentrations of vitamin E on peroxidation susceptibility was examined. 2. Laying hens (Ross 1 broiler-breeder strain) were maintained on diets containing either 147 (control diet) or 365 (high vitamin E diet) microg vitamin E/g feed. 3. In the day-16 embryo, the concentrations of of vitamin E in the yolk sac membrane, liver, brain and lung were respectively 5.0, 4.3, 1.7 and 5.6 times greater for those derived from the hens on the high vitamin E diet compared with those from the control group. 4. In the day-old chick, the concentrations of vitamin E in the yolk sac membrane, liver, brain and lung were respectively 14.8, 2.8, 3.0 and 5.1 times greater for those derived from hens on the high vitamin E diet compared with those from the control group. 5. Homogenates of tissues from the day-old chick were incubated in the absence and presence of Fe2+ in order to determine the extent of spontaneous and iron-stimulated peroxidation as measured by the generation of thiobarbituric acid reacting substances. For the chicks derived from hens on the control diet, the brain was markedly more susceptible to both spontaneous and iron-stimulated peroxidation than were the other tissues. Tissues from the chicks derived from the hens on the high vitamin E diet exhibited significantly reduced susceptibilities to peroxidation. In particular, the susceptibility of the brain was reduced to the same level as that of the other tissues. 6. It is concluded that the high peroxidative susceptibility of the chick's brain can be normalised by supplementation of the parent hen with vitamin E.     

Effects of a hydrolyzed collagen dressing on the healing of open wounds in dogs

OBJECTIVE: To determine the effects of a hydrolyzed bovine collagen dressing (HBCD) on healing of open wounds in healthy dogs. ANIMALS: 9 female Beagles. PROCEDURES: 2 full-thickness skin wounds were made bilaterally on the trunk of each dog. Wounds on 1 side were treated with powdered HBCD covered with a semiocclusive nonadherent bandage. Wounds on the other side (control wounds) were covered with a semiocclusive nonadherent bandage only. Wound healing was subjectively assessed, and percentage increase in tissue perfusion was assessed by use of laser Doppler perfusion imaging (LDPI). Planimetry was performed to determine the percentages of contraction, epithelialization, and total wound healing. Biopsy specimens were examined microscopically to evaluate histologic changes. RESULTS: The HBCD did not induce a strong inflammatory reaction, as reflected by results of LDPI and histologic examination. Moreover, HBCD appeared hydrophilic and provided an environment to keep wounds clean and enhance early epithelialization. After treatment for 7 days, treated wounds had a significantly greater percentage of epithelialization than control wounds (12.13 vs. 7.03%). CONCLUSION AND CLINICAL RELEVANCE: The hydrophilic property of HBCD may cleanse contaminated wounds with the body's homeostatic fluids and enhance early wound epithelialization.