Identification and validation of housekeeping genes as internal control for gene expression in an intravenous LPS inflammation model in chickens

Real-time PCR has become a powerful tool for the detection of inflammatory parameters, including cytokines. Reference or housekeeping genes are used for the normalization of real-time RT-PCR results. In order to obtain reliable results, the stability of these housekeeping genes needs to be determined. In this study the stability of five genes, including beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine phophoribosyl-transferase (HPRT), ubiquitin (UB) and glucose-6-phosphate dehydrogenase (G6PDH), was determined in a lipopolysaccharide inflammation model in chickens. beta-Actin appeared to be the most stable single gene in our model. Because the use of a single gene for normalization can lead to relatively large errors, the use of the geometric mean of multiple reference genes or normalization factor is preferred. The most stable combination for gene expression analysis in this lipopolysaccharide inflammation model in chickens is G6PDH and UB, since their correlation coefficients were 0.953 and 0.969, respectively (BestKeeper) and an M value of 0.34 and a low V(2/3) value of 0.155 (geNorm) were obtained. The use of HPRT and GAPDH should be avoided. The stable housekeeping genes, G6PDH and UB together, can be used to normalize the expression of pro-inflammatory cytokines in a lipopolysaccharide inflammation model in chickens.     

Cluster of cases of malignant schwannoma in cattle

Between 1998 and 2001, several cases of ataxia and paresis followed by recumbency and death were reported in cows from different farms in a restricted area of the Argentinian Patagonia. Five cases of this cluster were studied and a diagnosis of malignant schwannoma was established. Electron microscopy (em) of tumour samples from three of the animals revealed intracytoplasmic or interstitial structures resembling retroviral particles. Attempts to isolate a viral agent from the tumours were unsuccessful but the epidemiological data and the em findings suggest a viral aetiology.     

A Single Sample Method for Evaluating 51Chromium-Ethylene Diaminic Tetraacetic Acid Clearance in Normal and Hyperthyroid Cats

Background: Chronic kidney failure is frequently seen in middle-aged and elderly cats. ⁵¹Chromium-ethylene diaminic tetraacetic acid (⁵¹Cr-EDTA) clearance and single blood sample (SBS) method are used in several species to estimate the glomerular filtration rate (GFR).
Hypothesis: The hypothesis of this study was that ⁵¹Cr-EDTA clearance could be determined using an SBS method in normal and hyperthyroid cats.
Animals: Forty-six cats were included in this study, with an average age of 9.5 years. Of these cats, 27 had hyperthyroidism; 19 were healthy.
Methods: After IV injection of ⁵¹Cr-EDTA (average dose: 4.25 MBq), 7 blood samples were obtained between 5 and 240 minutes. Reference clearance was calculated in mL/min and mL/min/kg body weight, using a 2-compartment model. Optimal time for clearance measurement with SBS was then determined by systematically comparing each individual plasma concentration to the reference multisample clearance.
Results: The average reference plasma clearance of ⁵¹Cr-EDTA for all cats was 14.9 mL/min (3.7 mL/min/kg). The clearance in hyperthyroid cats averaged 16.4 mL/min (4.3 mL/min/kg) and in normal cats averaged 10.3 mL/min (2.4 mL/min/kg).
The optimal time for the SBS was 48 minutes after injection of tracer ⁵¹Cr-EDTA ( R ²= 0.9414), giving the following converting equation: clearance = (0.0066 × DV_{48 minutes}) – 0.9277 (in mL/min).
Conclusions and Clinical Importance: In this study, the single sample ⁵¹Cr-EDTA clearance method was used to estimate the global GFR in cats. The method identified differences in clearance between normal and hyperthyroid cats. The optimal time for an SBS was 48 minutes.     

Transmission of methicillin resistant Staphylococcus aureus among pigs during transportation from farm to abattoir

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in pigs at abattoirs is higher than in pigs sampled on farms. This study investigated whether MRSA negative pigs can become MRSA positive during transportation from the farm to the abattoir after exposure to other pigs and environmental sources of MRSA. Nasal swabs were collected from four batches of pigs during loading at the farm, on arrival at the abattoir and after stunning. Environmental wipes were taken from lorries after transporting pigs and from lairages after holding pigs. All pigs (n = 117) tested MRSA negative before transportation. On arrival at the abattoir, 12/117 (10.3%) pigs in two batches tested MRSA positive. In lorries that tested positive after transportation, the prevalence of MRSA positive pigs was 21.1%, whereas no MRSA was detected in pigs that had been transported in lorries that tested negative after transportation. At stunning, all batches and 70/117 (59.8%) pigs tested MRSA positive. Pigs can become MRSA positive in the short period of time during transportation from the farm to stunning at the abattoir.     

MicroRNA及其对哺乳动物繁殖的影响

卵巢功能的正常维持有赖于卵巢生殖细胞和体细胞之间的相互作用及若干卵巢自分泌和旁分泌的调节.这些调节物质调控卵巢内不同的细胞活动,包括细胞生长、分化和凋亡,从而对卵泡的发育起着至关重要的作用.Micro-RNAs(miRNAs)是一种小的、非编码、21～25 nt长的单链小分子RNA,近年来科学家们发现其在基因表达转录后水平的调控发挥了重要作用.鼠上的研究显示,miRNAs在卵母细胞成熟和卵巢卵泡发育过程中有调控功能.人上的研究也表明,miRNAs影响颗粒细胞中特定的基因表达,并参与卵巢癌的形成和发展.本文将对miRNAs的生物合成,及其在哺乳动物繁殖系统中的表达,正常和病理情况下miRNAs在繁殖系统中可能的调控作用作一综述.     

Studies on Haemonchus contortus. X. The effect of Trichostrongylus axei in Merinos on artificial pastures

A recently established Eragrostis curvula pasture 1,1 ha in size at Hennops River in the Transvaal Highveld was contaminated from 6 October 1976 until 20 May 1977 by 30 weaned Merinos, each infested with 7 000 infective larvae of H. contortus. On 2 November 1976 3 groups of weaned Merinos and again on 10 January 1977 a further group of Merinos were infested with 40 000 infective larvae of Trichostrongylus axei. From 19 November a single group of sheep, predosed with T. axei, and a control group grazed with the seeders for 8 weeks. After 6 weeks another group of sheep dosed with T. axei grazed with them, thus ensuring a 2-week overlap. This continued until autumn, when the last groups were removed. Efficacy against challenge reached Class C (greater than 50% effective against total worm burdens of H. contortus in greater than 50% of sheep) in early summer and autumn, but only reached significant levels of either P less than 0,01 or P less than 0,05 in summer. Peak worm burdens in controls were recorded in early autumn.     

Infarction of the pons and medulla oblongata caused by arteriolar thrombosis in a horse

Infarction of the pons and rostral medulla secondary to arteriolar thrombosis was documented histologically in a 17-year-old mixed-Arabian female horse. Clinically, the animal experienced a sudden onset of a head tilt and subsequent non-controllable seizures. There was no historical, clinical or histological evidence to suggest the presence of infection of Equine Herpesvirus-1 or the feeding of corn contaminated by Fusarium moniliforme.     

Expression of vascular endothelial growth factor and endothelial nitric oxide synthase is increased in the placenta of sheep at high altitude in the Andes

Fetal weight and the placenta of sheep at high altitude (HA) are affected by hypoxia. Placental changes (an increase in placental size and vascularization) are greater in ewes from populations that have lived for several generations at HA than in those exposed during just 1 gestation. This study investigated placental expression of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS), 2 molecules involved in placental angiogenesis that could be upregulated by hypoxia. Two groups of ewes were maintained at HA (3589 m) during pregnancy: HA-native ewes (group HH) and ewes native to lowlands but moved to HA immediately after the diagnosis of pregnancy (group LH). A control group (LL) was kept at sea level. Near term, placentomes were removed, weighed, and processed for immunohistochemical detection of VEGF and eNOS, as well as for vascular area measurement. Placental weight was significantly higher in the HH group than in the LH and LL groups; between the latter 2 groups there was no significant difference. The placental area occupied by vasculature was significantly greater in both the HA groups than in the LH group; the number of placentomes was greatest in the LL group. The density of VEGF and eNOS in the placentome tissue was significantly greater in both HA groups than in the LL group. Although the density of VEGF was significantly lower in the HH group than in the LH group, no differences were observed in eNOS density between the HH and LH animals. These results demonstrate that chronic hypoxia upregulates the expression of placental VEGF and eNOS, suggesting an important role of these molecules in the placental response to HA hypoxia. In addition, an attenuated response to hypoxia in VEGF synthesis may be part of the long-term process of adaptation to HA.     

Monitoring distances travelled by horses using GPS tracking collars

Objective The aims of this work were to (1) develop a low-cost equine movement tracking collar based on readily available components, (2) conduct preliminary studies assessing the effects of both paddock size and internal fence design on the movements of domestic horses, with and without foals at foot, and (3) describe distances moved by mares and their foals. Additional monitoring of free-ranging feral horses was conducted to allow preliminary comparisons with the movement of confined domestic horses. Procedures A lightweight global positioning system (GPS) data logger modified from a personal/vehicle tracker and mounted on a collar was used to monitor the movement of domestic horses in a range of paddock sizes and internal fence designs for 6.5-day periods. Results In the paddocks used (0.8–16 ha), groups of domestic horses exhibited a logarithmic response in mean daily distance travelled as a function of increasing paddock size, tending asymptotically towards approximately 7.5 km/day. The distance moved by newborn foals was similar to their dams, with total distance travelled also dependent on paddock size. Without altering available paddock area, paddock design, with the exception of a spiral design, did not significantly affect mean daily distance travelled. Feral horses (17.9 km/day) travelled substantially greater mean daily distances than domestic horses (7.2 km/day in 16-ha paddock), even when allowing for larger paddock size. Conclusions Horses kept in stables or small yards and paddocks are quite sedentary in comparison with their feral relatives. For a given paddock area, most designs did not significantly affect mean daily distance travelled.