Detection of a novel chloramphenicol resistance plasmid from "equine" Staphylococcus sciuri

A small chloramphenicol resistance (Cm) plasmid of 4.65 kB could be detected in an "equine" Staphylococcus sciuri-culture. This plasmid, designated as pSC3, was identified by interspecific protoplast transformation. On the basis of restriction endonuclease analyses a detailed restriction map of pSC3 could be constructed. This allowed structural comparisons of pSC3 with Cm-plasmids of other staphylococcal species from infections of humans and animals and identification of pSC3 as a member of the pC 221-family of staphylococcal Cm-plasmids. The pSC3-plasmid encoded an inducible chloramphenicol acetyltransferase as confirmed by enzymatic assays. This enzyme could be demonstrated in cell-free lysates of Cm-induced pSC3-transformants.     

Anesthetic management of a white rhinoceros (Ceratotherium simum) undergoing an emergency exploratory celiotomy for colic

ObservationsA 26-year-old male white rhinoceros (Ceratotherium simum), weighing approximately 2000 kg was anesthetized for an exploratory celiotomy. Sedation was achieved with intramuscular butorphanol (0.04 mg kg^?1) and detomidine (0.025 mg kg^?1) and induction of anesthesia with intravenous glyceryl guaiacolate (50 g) and three intravenous boluses of ketamine (200 mg, each); the trachea was then intubated and anesthesia maintained with isoflurane in oxygen using a circle breathing system. Positioning in dorsal recumbency for the surgery and later in sternal recumbency for the recovery represented challenges that added to the prolonged anesthesia time and surgical approach to partially correct an impaction. The rhinoceros recovered uneventfully after 10.4 hours of recumbency.ConclusionsAnesthetic management for an exploratory celiotomy with a midline approach is possible in rhinoceroses, although planning and extensive staff support is necessary to adequately position the patient.     

The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.     

A three‐dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two‐dimensional (2D) and three‐dimensional (3D) conditions. The 3D was performed in six‐well AlgiMatrix? (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50–125 µm for CC and 175–200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases‐1, ‐2, ‐9 and ‐13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E‐cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)‐derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Breeding objectives of Brangus cattle in Brazil

Our goal was to define a breeding objective for Brangus beef cattle in Brazil. Bioeconomic models were produced and used to estimate economic values (EVs). The scenarios simulated were typical full-cycle beef production systems that are used in tropical and subtropical regions. The breeding objective contained pregnancy rate (PR), warm carcass weight (WCW), mature cow weight (MCW), number of nematode eggs per gram of faeces (EPG) and tick count (TICK). Two models were used in series to estimate the EV. A deterministic model was used to simulate effects of PR, WCW and MCW on profitability with a constant parasite load. Subsequently, stochastic models were used to estimate economic values for TICK and EPG as consequences of their environmental effects on weight gains, mortality and health costs. The EV of PR, WCW, MCW, EPG and TICK, was US$1.59, US$2.11, −US$0.24, −US$5.35 and −US$20.88, respectively. Results indicate positive emphasis should be placed on PR (12.49%) and WCW (65.07%) with negative emphasis on MCW (13.92%), EPG (2.77%) and TICK (5.75%). In comparison with the indexes usually used, these results suggest a reformulation in the selection indexes of the beef production system in tropical and subtropical regions in order to obtain greater profitability.     

Cytoprotection by neutral fraction of tannat red wine against oxidative stress-induced cell death

Some of the beneficial effects of the Mediterranean diet on human pathologies have been attributed to red wine polyphenols. It has been postulated that the antioxidant activity of the latter would be also responsible for the cytoprotective capacity of red wine that has been reported in a few papers. Nevertheless, red wine shows a complex composition, and the active fraction is not known yet. In this context, the protective capacity of total lyophilized extracts of red wine and anthocyanin, neutral, or acidic fractions, was explored in PC12 cells in culture after a hydrogen peroxide insult. Although all fractions showed high antioxidant activity, only the neutral fraction was cytoprotective. The analysis of this active fraction showed that it was rich in the aglycons quercetin and myricetin as well as the glycosides of kaempferol, isorhamnetin, epicatechin, and catechin, some of which are known to be cytoprotective. This is the first paper to reveal the active fraction of total wine responsible of its cytoprotection.     

Montenegro's skin reactions and antibodies against different Leishmania species in dogs from a visceral leishmaniosis endemic area

In this study, humoral (circulating anti-Leishmania antibodies) and cellular (Montenegro's skin test) immune responses of dogs from an endemic area of visceral leishmaniosis were tested using Leishmania chagasi, Leishmania amazonensis and Leishmania braziliensis antigens. The antibody response was tested in three animal groups, selected according to their anti-L. chagasi antibody activity, as measured by ELISA in the serum: 19 negative (O.D. below 0.30), seven with undefined (O.D. between 0.40 and 0.70) and 12 positive (O.D. above 1.0) ELISA result. In the group of animals with positive ELISA, the antibody activity against L. chagasi antigens (mean O.D.=1.31) was significantly higher (ANOVA, P<0.01) than against L. amazonensis (mean O.D.=0.88) or L. braziliensis (mean O.D.=0.87) antigens. The Montenegro's skin test results obtained with L. chagasi and L. braziliensis antigens showed a fair agreement (kappa=0.309). The same was observed when antigens from L. braziliensis and L. amazonensis were compared (kappa=0.374), whereas a moderate agreement between the results from tests performed with L. chagasi and L. amazonensis antigens was observed (kappa=0.530). The induration areas obtained with L. braziliensis antigen were smaller than those obtained with the other antigens. The data presented herein indicate that the use of antigens from different Leishmania species may interfere with the results of the immunological tests performed in dogs in an endemic area of visceral leishmaniosis.     

Development of colchicine‐induced tetraploid St. Augustinegrass (Stenotaphrum secundatum) lines

St. Augustinegrass is well suited for lawns and commercial landscapes. While many genotypes are cross‐fertile, all cultivars are propagated vegetatively in sod production. To ensure varietal purity, development of sterile triploid hybrids by crossing tetraploid and diploid genotypes has been successfully used in other warm‐season turfgrasses. Applying this model in St. Augustinegrass would be beneficial to sod producers and turf managers who require purity for certification and uniformity for performance, respectively. This study was conducted to develop colchicine‐induced tetraploid lines of St. Augustinegrass. Seeds of cultivar ‘Raleigh’ were treated with four colchicine concentrations at four exposure times. A non‐treated control was included among the treatments. Seedlings that germinated were screened for genome size changes using flow cytometry. Line DSA 13005 and two progeny lines derived through selfing, DSA 16001 and DSA 16016, were corroborated as tetraploids (2n = 4x = 36) through chromosome counts. These lines will be used in future breeding efforts to attempt development of sterile triploid cultivars of St. Augustinegrass.