Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

In-hospital development of an aorto-cardiac fistula in a Warmblood gelding with chronic renal disease

A 20-year-old Warmblood gelding presented for evaluation and treatment of ventral oedema and azotaemia of unknown aetiology. On presentation, a diastolic heart murmur was appreciated and echocardiography revealed moderate aortic insufficiency due to chronic degenerative valve disease. The horse was hospitalised and failed to respond to oral and i.v. fluids and diuretics. Following discontinuation of all fluid and diuretic therapy, the horse became acutely agitated and developed monomorphic ventricular tachycardia. The ventricular tachycardia spontaneously converted to normal sinus rhythm, however the heart murmur changed in timing to a right basilar continuous murmur and bounding jugular pulses were noted. Repeat echocardiogram revealed an aorto-cardiac fistula with dissection into the basilar interventricular septum and left-sided chamber volume overload that was not previously present. Attempts at stabilisation were unsuccessful and euthanasia was elected. Post-mortem examination confirmed chronic renal disease of unknown aetiology in addition to an aorto-cardiac fistula originating from the right sinus of Valsalva with subsequent dissection into the basilar interventricular septum.     

物候期对放牧牦牛瘤胃液、牧草中脂肪酸及乳脂中共轭亚油酸组成的影响及其相关性分析

为研究牧草物候期对其脂肪酸组成和放牧牦牛乳中共轭亚油酸（CLA）含量的影响及其相关关系,本研究在甘肃省甘南牧区选取平均体重为（234.9±10.5） kg的10头带犊甘南牦牛作为研究对象（自然放牧）,分别于牧草返青期、盛草期、枯草期采集试验牦牛采食牧草、乳和瘤胃液,分别测定瘤胃液、牧草中脂肪酸及乳脂中共轭亚油酸组成,并分析其相关关系,以期探明牧草物候期对牦牛瘤胃液和牧草中脂肪酸组成与放牧牦牛乳中共轭亚油酸含量的影响及其变化规律。结果表明：物候期显著影响了牧草中月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、亚油酸、亚麻酸、总脂肪酸、多不饱和脂肪酸、多不饱和脂肪酸/总脂肪酸（P<0.05）,盛草期牧草中月桂酸、肉豆蔻酸、棕榈酸、亚油酸、亚麻酸、总脂肪酸、多不饱和脂肪酸、多不饱和脂肪酸含量/总脂肪酸均显著高于返青期和枯草期（P<0.05）;物候期显著影响了牦牛乳中共轭亚油酸异构体c9t11、共轭亚油酸异构体t10c12和共轭亚油酸总含量（P<0.05）,盛草期放牧牦牛乳中共轭亚油酸异构体c9t11、共轭亚油酸异构体t10c12和共轭亚油酸总含量显著高于返青期和枯草期（P<0.05）;物候期显著影响了牦牛瘤胃pH值、乙酸、丙酸、乙酸/丙酸、异丁酸、丁酸、异戊酸、戊酸、总挥发酸和反式11-油酸含量（P<0.05）,盛草期牦牛瘤胃乙酸、丙酸、丁酸、戊酸总挥发酸和反式11-油酸含量显著高于返青期和枯黄期（P<0.05）;牧草粗蛋白、粗脂肪、无氮浸出物、单宁酸、总脂肪酸和瘤胃液pH值与放牧牦牛乳中CLA含量显著正相关（P<0.05）,粗纤维和粗灰分与放牧牦牛乳中CLA含量显著负相关（P<0.05）,多不饱和脂肪酸（PUFAs）和瘤胃总挥发酸（TVFA）与放牧牦牛乳中CLA含量无显著相关关系（P>0.05）。     

山东省菏泽市某规模猪场不同猪群伪狂犬病野毒感染情况调查

为了解规模猪场不同猪群的伪狂犬病病毒(PRV)感染情况及场区病毒载量分布特点,在山东省菏泽市某规模猪场,利用实时荧光定量PCR方法,对该猪场不同孕龄母猪、不同阶段生长猪群,以及各生产阶段场区环境,采集猪鼻拭子和场区环境拭子进行PRV-gE核酸检测,并对检测结果进行统计分析。结果显示:在294份猪鼻拭子中,检出42份PRV-gE核酸阳性,总阳性率为14.28%;母猪群PRV-gE阳性率为17.83%(23/129),且妊娠前后各阶段阳性率呈先上升后下降趋势,中期较高(24.00%),但差异不明显(P>0.05);不同阶段生长猪群的平均PRV-gE阳性率为11.52%(19/165),随日龄增加,阳性率也呈先上升后下降趋势,80~90日龄育肥猪最高(28.13%),与其他生长阶段猪群差异明显(P<0.05);除饲料、水源、上猪台和出猪台外,其他大部分场区环境均检测到PRV-gE核酸阳性;育肥猪群病毒载量最高,为3.6×105拷贝数/mL,其猪舍环境的病毒载量也较高,与其他环境及生长阶段猪群差异均明显(P<0.05)。结果表明,不同猪群包括免疫猪群均可遭受PRV野毒感染,尤其是母猪妊娠中期和猪育肥阶段早期,且大部分场区环境均可被污染。结果提示,规模猪场要加强各种猪群尤其是育肥猪群的伪狂犬病免疫,通过监测来调整和优化免疫程序,同时要加强生物安全,防止病毒传入和扩散。     

Randomly primed,strand-switching,MinION-based sequencing for the detection and characterization of cultured RNA viruses

RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.     

宁夏地区荷斯坦牛成母牛淘汰情况及长寿性影响因素分析

试验通过收集出生、产犊和离群的记录,探究宁夏地区荷斯坦牛成母牛的淘汰情况,分析影响该地区奶牛长寿性的因素。对15 523头荷斯坦牛的生产寿命、在群寿命和利用胎次进行描述性统计,分析淘汰时的胎次、季节和泌乳阶段的分布情况;计算生产寿命、在群寿命和利用胎次之间的表型相关,利用固定模型分析场-出生年、出生季节、牧场规模、淘汰原因和头胎产犊月龄对长寿性的影响。结果显示,宁夏地区荷斯坦牛的平均利用胎次为2.36胎,平均生产寿命为736.59 d。荷斯坦牛成母牛在产后第1个月内的淘汰风险最高,随着泌乳阶段的延长,成母牛在每个胎次内的淘汰风险均逐渐降低;随着淘汰胎次的增加,奶牛淘汰更加集中发生在产后泌乳早期;生产寿命、在群寿命和利用胎次之间存在较高的表型相关（均>0.9）,场-出生年、出生季节、牧场规模、淘汰原因和头胎产犊月龄对长寿性有显著影响（P<0.05）,春季出生和22月龄头胎产犊的奶牛长寿性表现更好。本研究初步揭示了宁夏地区荷斯坦牛成母牛的淘汰规律和长寿性的影响因素,为宁夏地区牛群选育成母牛长寿性状奠定了基础。     

Socio-spatial organization reveals paternity and low kinship in the Honduran white bat (Ectophylla alba) in Costa Rica

Ectophylla alba is a tent-making bat that roosts in mixed-sex clusters comprising adults and offspring. Our goal was to determine the genetic identity of individuals belonging to different roosting groups. We tested the hypothesis of kin selection as a major force structuring group composition. We used 9 microsatellites designed for E. alba to determine the genetic identity and probability of parentage of individuals. We analyzed parentage and kinship using the software ML-Relate, GenAIEx, and Cervus. The obtained relationship probabilities (0.5) revealed a clear maternal relationship between female adults and offspring with allele compatibility, and at least 5 relationships between male adults and pups. We found a low degree of relatedness within roosting groups. Between roosting groups at different sites, the mean probability of a half-sibling relationship ranged from 0.214 to 0.244 and, for full-sibling relationship, from 0.383 to 0.553. Genetically, adult individuals were poorly related within clusters, and kinship as an evolutionary force could not explain group membership.     

Estimation of Runoff,Peak Discharge and Sediment Load at the Event Scale in a Medium‐Size Mediterranean Watershed Using the Annagnps Model

In this paper, the Annualized Agricultural Non‐Point Source (AnnAGNPS) model has been used to estimate runoff, peak discharge and sediment load at the event scale in a Mediterranean watershed. The study area is the Carapelle torrent, Southern Italy (area = 506 km²), where continuous rainfall, streamflow and sediment load data are available. Nineteen flood events have been registered in the period 2007–2009 and were used for the application of the model. The aim of the paper is to evaluate the predictive accuracy of the model at the event scale, in a medium‐size watershed, given the specific conditions of the semi‐arid environments. A sensitivity analysis has been carried out to assign the correct parameterization: the mean normalized output variation of the most meaningful input parameters pointed out the influence of the curve number on runoff, peak discharge and sediment load predictions (values greater than 1); the MN Manning's roughness coefficient and K, C and P factors of the universal soil loss equation showed a moderate influence on sediment load simulations (values between 0·5 and 1). The selection of the Soil Conservation Service synthetic storm types has been based on the observed storm events analysis to improve the peak discharge simulations. The model prediction has proved to be good for runoff (R² = 0·74, NSE = 0·75, W = 0·92) and peak discharge (R² = 0·85, NSE = 0·70, W = 0·94), and satisfactory for sediment yield (R² = 0·70, NSE = 0·63, W = 0·91). The relative error is lower for high events; this result is quite interesting in semi‐arid environments, where most of the annual sediment yield is concentrated in a few, severe events. Copyright © 2013 John Wiley & Sons, Ltd.