绵羊DUSP6基因多态性及其与产羔数关联分析

文章旨在探究绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T、g.125589006G>A四个位点多态性及其与产羔数之间的关系,以期找到与绵羊高繁殖力相关的分子标记。利用全基因组重测序结合Sequenom MassARRAY誖SNP技术对多羔绵羊品种(小尾寒羊、湖羊、策勒黑羊)和单羔绵羊品种(滩羊、苏尼特羊、萨福克羊、草原型藏羊)DUSP6基因上述4个多态位点进行检测,并与小尾寒羊产羔数进行关联分析。结果表明:绵羊g.125589716G>A位点在多羔绵羊品种中存在AA、GA、GG三种基因型,在单羔绵羊品种中存在GA、GG两种基因型;g.125587728C>T、g.125589714C>T位点在单、多羔绵羊品种中均存在CC、CT、TT三种基因型;g.125589006G>A位点在单、多羔绵羊群体中均只存在GA、GG两种基因型。绵羊DUSP6基因g.125589716G>A位点基因型频率在单、多羔绵羊品种之间差异显著(P<0.05),等位基因频率在单、多羔绵羊品种间差异极显著(P<0.01);g.125587728C>T、g.125589714C>T位点基因型频率和等位基因频率在单、多羔绵羊品种间差异均达极显著水平(P<0.01);g.125589006G>A位点基因型频率和等位基因频率在单、多羔绵羊品种间差异不显著(P>0.05)。关联分析表明,DUSP6基因各SNPs多态性与小尾寒羊不同胎次产羔数之间无显著关联(P>0.05)。综上说明,绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T以及g.125589006G>A等4个位点的多态性与小尾寒羊各胎次产羔数之间均无显著关联(P>0.05),不适用于小尾寒羊多羔性状选育。     

Interleukin-6 and insulin incrase and nitric oxide and adiponectin decrease in blind dogs with pituitary-dependent hyperadrenocorticism

In this study, two populations of dogs with pituitary dependent hypercortisolism (PDH) were compared over a 2-year period. One group had normal vision (Group A, n=27) and one group was blind (Group B, n=20). Group B was characterised by the rapid appearance of the clinical signs of PDH that precede blindness. We found increases in pre-adrenocorticotropic hormone cortisol (P=0.002), IL-6 (P=0.0001), insulin, and insulin sensitivity (detected with the Homeostatic Model Assessment, P<0.0001) in Group B but not in Group A. The nitric oxide (NO) and the total adiponectin concentrations decreased (P=0.0001 and P=0.02, respectively) in Group B versus Group A. The IL-6 and insulin concentrations and the HOMA-A index were positively correlated with the cortisol concentration and were negatively correlated with the NO concentration. With the exception of adiponectin, the other variables were associated with blindness. We concluded that blindness in PDH is a haemodynamic event associated with metabolic changes, with the increase in the IL-6 concentration and the decrease in the NO concentration affecting the retinal vasculature and producing a high risk of vision loss.     

Liveweight gain and copper status of young deer treated or untreated with copper oxide wire particles on ten deer farms in Canterbury

AIMS: To measure the liveweight response of young deer to copper supplementation on a range of deer farms in the Canterbury region of the South Island, New Zealand, and relate any response to plasma copper concentration. METHODS: On each of 10 farms throughout north and mid-Canterbury, young deer (4-7 months old) were allocated by liveweight to two groups of 45-50. The deer in one group each received 5 g copper oxide wire particles (COWP) at the start of the project (March-June 2000) and a further 10 g COWP 3 months later (+Cu group), while the other group received no copper supplementation (-Cu group). Liveweight (+/-1 kg) of all deer was recorded monthly. A 10 ml blood sample was taken from a sub-sample of 10 animals in each group at the beginning of the trial and from the same animals 3 and 6 months later for measurement of plasma copper concentration. RESULTS: There was no significant effect (p=0.96) of treatment with COWP on liveweight gain (overall mean liveweight gain = 169 g/day). This result was consistent for all farms and for both periods. Mean plasma copper concentration declined from 12.5 (SD 3.4) to 10.3 (SD 3.0) micromol/l in +Cu groups, and to a significantly lower level (p<0.001) of 8.6 (SD 3.2) in -Cu groups after 3 months, at which time 38% of -Cu animals were considered hypocupraemic (<8 micromol/l). At the end of the second 3-month period there was no significant treatment effect on plasma copper concentration, which averaged 9.4 (SD 2.3) micromol/l. Of the feed samples submitted (n=46), only 17% had a copper concentration <5.0 mg/kg dry matter (DM), 13% had a molybdenum concentration >1.0 mg/kg, and 21% had a sulphur content >3.5 g/kg. CONCLUSIONS: Under the conditions of these trials, no response in liveweight gain to copper supplementation was observed despite evidence of hypocupraemia in 38% of -Cu animals, which gained weight at similar rates to those that had adequate plasma copper levels. The extent of the hypocupraemia was either not sufficiently severe, or not maintained for a long enough period to cause copper deficiency resulting in reduced liveweight gain. No other signs of copper deficiency were evident. There is scope for deer farmers to reassess the need for copper supplementation in young deer.     

Use of computer tomography for imaging of Crassicauda grampicola in a Risso's dolphin (Grampus griseus).

A mature male Risso's dolphin (Grampus griseus) stranded along the coasts of Friuli Venezia Giulia, northeast Italy, in May 2001. Parasitic infection with Crassicauda grampicola is often found in the tympanic bullae and pterygoid sinuses in many of the Risso's dolphins examined from the same area. For this reason, it was decided to perform computed tomography of the head to assess this imaging technique for the diagnosis of crassicaudosis in dolphins. A full postmortem examination confirmed the pathologic findings of the computed tomography scan. This technique can be considered a useful adjunct in the diagnosis of cranial crassicaudosis in live dolphins.     

Colocalization of DNA fragmentation and caspase-3 activation during atresia in pig antral follicles

Apoptosis is the cellular mechanism of ovarian follicular atresia. The major downstream effector of this phenomenon in many tissues is caspase-3 but little is known about its role in pig ovarian apoptosis. In the present study, we detected the localization of caspase-3 in parallel with nuclear fragmentation (TUNEL) on healthy and early atretic antral follicles. In healthy antral follicles caspase-3 and TUNEL positivity were occasionally recorded within theca layer. The incidence of DNA fragmentation, as indicated also by the biochemical detection, increased mainly in the granulosa layer of early atretic follicles. Quantitative analysis revealed, besides, that atresia was accompanied by a higher incidence of caspase-3 (57.20 +/- 20.05 versus 3.64 +/- 0.61 positive cells in atretic versus healthy follicles, respectively; P < 0.05), of TUNEL positivity (20.13 +/- 9.33 versus 0.42 +/- 0.12; P < 0.05) and simultaneous immunostaining for caspase-3 and TUNEL (15.02 +/- 6.95 versus 0.31 +/- 0.05; P < 0.05) in the granulosa layer. In detached granulosa cells isolated from the follicular fluid of early atretic follicles a further significantly increase was recorded in the percentage of TUNEL positivity and in the incidence of cells that showed colocalization of caspase-3 activity and DNA fragmentation. Granulosa cells of early atretic follicles exhibited a higher positivity for caspase-3 localized in the cytoplasm and occasionally in the nucleus area of granulosa cells. These results indicate that capsase-3 was involved and precociously activated during the process of atresia. Finally, the progressively higher incidence of TUNEL positivity and of double immunostaining in atretic cells collected within the follicular fluid seems to indicate that proteases activity leads only tardily in a detectable DNA fragmentation.     

Clinical nutrition in gerontology: chronic renal disorders of the dog and cat

The recent advances in the nutrition of companion animals has resulted in a longer possible life-span for dogs and cats and an improvement in their quality of life. Numerous studies about geriatric animals show that an aging dog or cat requires a specific nutritional formulation that considers the metabolic changes associated with age. A correct diet plays an important role in the treatment of some chronic pathologies in aging animals, particularly those for which the aging process modifies the organ function. A correct diet can provide therapeutic support to the administration of drugs that can sometimes compromise organ function. In the present study, we identify key aspects of the clinical nutrition during chronic renal disorders of dogs and cats, diseases with an elevated incidence and a major cause of mortality in geriatric animals. The aim of nutritional treatment for dogs and cats affected by chronic renal disorders is to improve the quality and length of life, assuring an adequate amount of energy and slowing the progression of renal failure. To improve treatment efficacy it is necessary to prepare different dietary rations during the various stages of disease, on the basis of clinical signs and laboratory data.     

Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Clinical, humoral and IFNgamma responses of cattle to infection with Mycoplasma mycoides var. mycoides small colony and attempts to condition the pathogenesis of the infection

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides var. mycoides small colony (MmmSC), is one of the most important diseases of cattle in Africa. The role of innate or acquired cell mediated and humoral immunity in conferring protection against MmmSC infection has not yet been elucidated. On the other hand, the pathological lesions caused by the aetiological agent have been considered indicative of an immunopathological process. In this study ten na?ve cattle were exposed to in-contact infection with animals infected by intubation with a strain of MmmSC. Clinical signs, antibody response, IFNgamma release and pathological changes at necropsy were analysed and compared with the events following in-contact infection of an equal number of animals kept under daily treatment with cyclosporine for the entire observation period of 84 days. Cyclosporine is a suppressor of the immune response related to the T-cell system. Under the conditions of the experiment, cyclosporine appeared to condition the pathogenesis of CBPP by delaying the events that follow infection, bringing further support to the possibility that the immune response may have an impact on the disease outcome.     

实时定量PCR技术在瘤胃微生物定量分析中的研究应用

随着对瘤胃微生物研究的逐步深入,一些以分子生物学技术为基础的定量分析方法对瘤胃微生物的研究方兴未艾。本文简要介绍了实时定量PCR技术的原理、操作程序以及该技术在反刍动物瘤胃微生物定量分析中的应用现状、取得的成果和前景展望。     

Evidence of West Nile virus lineage 2 circulation in Northern Italy

A West Nile virus (WNV) strain belonging to lineage 2 was for the first time detected in two pools of Culex pipiens collected in the province of Udine and in tissues of a wild collared dove (Streptopelia decaocto) found dead in the province of Treviso, in North East of Italy. It was molecularly identified by group and WNV lineage specific RT-PCRs and characterized by partial sequencing of the NS3 and NS5 genes. When compared with the sequences of same fragments of NS3 and NS5 of the WNV lineage 2 strain isolated from birds of prey in Hungary (2004), the phylogenetic analysis of these sequences revealed 100% and 99% similarity, respectively. As the Hungarian strain, the NS3 selected sequence differed from the 2010 Greek isolate by one amino-acid located at 249 site which is the site involved in genetic modulation of WNV pathogenicity. The Italian and Hungarian strains have histidine rather than proline at this site. The presence of a lineage 2 strain in regions where the lineage 1 strain is still circulating, creates a new scenario with unpredictable consequences. In this situation comprehensive investigations on the occurrence, ecology, and epidemiology of these different WNV strains circulating in Italy become the highest priority.