Maternal undernutrition during the pre- and post-conception periods in twin-bearing hairsheep ewes: effects on fetal and placental development at mid-gestation

To evaluate the effects of pre- and post-conception undernutrition (UN) on fetal and placental development at mid-gestation, 28 Katahdin × Pelibuey multiparous ewes were blocked by weight and assigned to the following four dietary treatments (n = 7 each): ewes fed 100% (control) or 60% of their nutritional requirements 30 days before mating (UNPre), 50 days after mating (UNPost) or during both periods (UNB). Four twin-bearing ewes were selected per treatment at day 50 post-conception and then slaughtered at day 75 of gestation to analyze their fetuses. Control fetuses were heavier (P < 0.05) than UNPost and UNB fetuses in 14.6 and 9.4%, respectively. Organ weights as percentage of the fetal weight (except for liver) and morphometric measurements (except for abdominal girth) were similar between control and UN fetuses (UNPre, UNPost, and UNB). Placental mass was heavier (P < 0.05) in control ewes than UNB ewes, but not relative to ewes of other treatments. The number of placentomes per ewe and placental efficiency were unaffected by UN treatments. Compared to control, only UNB ewes exhibited variations (P < 0.05) in the proportion of placentomes, specifically for type A (+13.8%) and B (?12.6%). Placentomes of type A and B had lower weight, length, and width of placentas in UNPost and UNB ewes than placentas of control ewes (P < 0.05). Overall results indicate that fetal and placental development of ewes carrying twins is mainly altered when nutritional restriction occurs simultaneously before conception and during the first third of pregnancy.     

Characterization of a Streptococcus equi ssp. equi Isolate From a Strangles Outbreak in Thailand

Strangles, which is caused by Streptococcus equi ssp. equi, is one of the major infectious respiratory diseases in horses. Knowledge of isolates from different areas of the world is important for investigating the different strains of the disease. In contrast to many other countries, currently little is known about S. equi ssp. equi isolates in Thailand. In 2014, a farm in Thailand imported 20 horses from Europe. Approximately 1 month after arrival, 50% of the horses had developed pyrexia, mucopurulent nasal discharge, and abscesses of the mandibular lymph nodes. Nasal swabs of mucopurulent discharge were sent to a diagnostic laboratory, and two isolates of S. equi ssp. equi were identified. One of the isolates was further characterized using seM gene polymerase chain reaction and sequence analysis. The seM sequence was then compared to the database of PubMLST-seM. It was found to contain SeM allele 48, an allele isolated from horses in the United Kingdom in 2006 and 2010. This result demonstrates the usefulness of SeM allele identification as a tool for investigating the source of related strains and for the epidemiologic study of strangles. To the best of the authors' knowledge, this is the first report of the identification of an SeM allele of the S. equi ssp. equi isolate in Thailand.     

Dissemination of the superantigen encoding genes seeL, seeM, szeL and szeM in Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus

Bacterial superantigens are one of the major virulence factors produced by Streptococcus pyogenes and Staphylococcus aureus. The two novel superantigen encoding genes seeM and seeL were described for S. equi subsp. equi which is known as the causative agent of strangles in equids. In the present study previously characterized S. equi subsp. equi strains and strains of various other animal pathogenic streptococcal species and subspecies were investigated for the presence of the superantigen encoding genes seeM and seeL by polymerase chain reaction. According to these studies seeL and seeM appeared to be a constant characteristic of all investigated S. equi subsp. equi strains. Surprisingly, one S. equi subsp. zooepidemicus strain (S.z. 122) was also positive for both genes. The species identity of this S. equi subsp. zooepidemicus strain could additionally be confirmed by sequencing the 16S rRNA gene and the 16S-23S rDNA intergenic spacer region. The superantigen encoding genes could not be found among additionally investigated S. equi subsp. zooepidemicus strains or among strains of seven other streptococcal species. The seeL and seeM genes of the S. equi subsp. equi strain S.e. CF32 and the genes szeL and szeM of the S. equi subsp. zooepidemicus strain S.z. 122 were cloned and sequenced. A sequence comparison revealed a high degree of sequence homology between seeL, szeL, speL and seeM, szeM and speM, respectively. The superantigenic toxins L and M seemed to be widely distributed virulence factors of S. equi subsp. equi, rare among S. equi subsp. zooepidemicus but did not occur among a number of other animal pathogenic streptococcal species.     

Eustachian tube diverticulum chondroids and neck abscessation in a case of Streptococcus equi subsp. equi

A 12-year-old, grey, crossbred pony mare was presented with a swelling in the neck over the area of the 2nd cervical vertebra (C2), which was found to be painful on palpation. The neck was held stiffly. Radiography of the cervical region showed a focal area of increased radio-opacity over the dorsal, caudal and lateral aspect of the dorsal spinous process of C2. Ultrasound confirmed the presence of a hypoechoic area approximately 15 cm in diameter superimposed over the dorsal spinous process of C2. An aspirate was taken of the mass, which revealed purulent exudate confirming the diagnosis of an abscess. The abscess was lanced with a scalpel blade and samples of the purulent material revealed a pure culture of Streptococcus equi subsp. equi. The guttural pouches (Eustachian tube diverticulae (ETD)) were then evaluated endoscopically and multiple chondroids were seen filling most of the right ETD. Surgery was subsequently performed and 189 chondroids were removed via a right-sided hyovertebrotomy. The ETDs were flushed and penicillin installed into both ETDs on 3 different occasions via a catheter introduced using a fibre optic scope. This procedure was repeated until a negative culture status was achieved in order to eliminate the carrier status.     

Effect of nutrient intake during pregnancy on fetal and placental growth and vascular development

Remarkable diversity of size and health of offspring exists after normal pregnancies. When pregnancies are complicated by an extrinsic variable such as inappropriate maternal nutrition, birth weight and health of the neonate are substantially affected. The placenta is the organ through which respiratory gases, nutrients, and wastes are exchanged between the maternal and fetal systems. Thus, transplacental exchange provides for all the metabolic demands of fetal growth. Transplacental exchange is dependent upon uterine and umbilical blood flow, and blood flow rates are in turn dependent in large part upon vascularization of the placenta. Therefore, factors that influence placental vascular development will have a dramatic impact on fetal growth and development, and thereby on neonatal mortality and morbidity. Recent work from our laboratories has focused on the effects of nutrient intake during pregnancy on placental growth and vascular development. Both nutrient restriction of the adult dam and overnourishment of the adolescent dam during pregnancy suppress placental cell proliferation and vascularity. Furthermore, placental expression of angiogenic factors and their receptors, factors that are known to affect vascular growth, are perturbed by level of nutrition. Studies in this area will lead to improved methods to manage nutritionally-compromised pregnancies.     

Bacteriological and Molecular Detection of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in Equines of Northern India

Present study was undertaken to study the prevalence of β-haemolytic streptococci in equine of northern temperate region of Jammu and Kashmir, India. One hundred and forty one samples were collected in duplicate from nasopharyngeal tract of diseased (53) and apparently healthy equine (88) for isolation and direct PCR. A total of 77 isolates of streptococci were recovered from 141 samples with an overall prevalence rate of 54.60%. Out of these 77 isolates, 52 were from diseased and 25 from apparently healthy animals. Of the 77 isolates, 4 were identified as Streptococcus equi subsp. equi, 56 as S. equi subsp. zooepidemicus and 17 as S. dysgalactiae subsp. equisimilis. Thus the overall prevalence of S. equi subsp. equi, S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis was 2.83, 39.71 and 12.05% respectively. The sensitivity of the PCR for the detection of S. equi species was found higher when attempted from direct swab samples.     

金银花、连翘及其药对对马链球菌马亚种耐药基因和毒力基因的影响

【目的】 研究金银花、连翘及金银花-连翘药对(金银花-连翘1∶1)对北疆地区携带fneB毒力基因的马链球菌马亚种(Streptococcus equi subsp. equi, SEE)耐药基因和毒力基因的影响。【方法】 首先对1 g/mL的金银花、连翘及金银花-连翘药对(金银花-连翘1∶1)水提物进行中药配比浓度梯度试验, 然后与携带fneB毒力基因的L1菌株、lytA+fneB+ply毒力基因的D1菌株和qnrA+bla_TEM+fneB基因的Y1菌株3种SEE菌株共培养, 检测中药对SEE菌株耐药基因bla_TEM、qnrA及毒力基因ply、fneB的影响; 将192只SPF级昆明小鼠均分为16组: 空白对照组(生理盐水)、金银花组、连翘组、金银花-连翘1∶1组、阴性对照组(Y1、L1和D1菌株组)及3种菌株分别与金银花、连翘和金银花-连翘1∶1共培养组, 各组药物或菌液经腹腔注射0.5 mL进行小鼠体内抑菌试验, 检测药物对小鼠的致病性和fneB毒力基因的影响。【结果】 中药最适配比浓度为中药水提物∶THB培养基∶待测菌液(D_{600 nm}值均为0.6)为1 000 μL∶500 μL∶20 μL; 与中药水提取物共培养的3株SEE菌株均未检出耐药基因和毒力基因, 且菌株形态结构均未发生改变。小鼠致病性试验结果显示, 阴性对照组的小鼠成活率分别为16.7%、8.3%和0;而L1+连翘、L1+金银花共培养组小鼠存活率分别为83.3%、75.0%, 金银花-连翘1∶1药对与3株SEE菌株共培养组小鼠成活率分别为41.7%、16.7%、50.0%;小鼠病理解剖结果显示, 除接种SEE菌株的小鼠肝脏肿大淤血、边缘钝圆外, 其余各组肝脏均正常。小鼠体内fneB毒力基因检测结果显示, Y1+金银花、Y1+连翘、Y1+金银花-连翘1∶1、L1+金银花、L1+金银花-连翘1∶1、D1+金银花、D1+连翘、D1+金银花-连翘组均携带fneB毒力基因, L1+连翘组未检出fneB毒力基因, 表明小鼠体内SEE菌株有重新获得fneB毒力基因的能力, 出现菌种反毒复壮, 其机制有待进一步研究。【结论】 金银花、连翘及金银花-连翘药对能够减弱SEE菌株的毒力基因和耐药基因, 从而对马腺疫疾病的防治具有指导意义, 为减抗、替抗提供理论支撑。     

The effects of endometrial damage on placental and fetal development in a mare

A 13-year-old ex-polo pony maiden mare with an extensively damaged endometrium was initially used as an embryo donor in a research herd. Embryos recovered from the mare were of normal morphology and size. Subsequently, the mare was inseminated and left pregnant. Early conceptus development was normal and she foaled a small (22 kg) but healthy filly after a gestation of 360 days. Examination of the placenta showed that development of the microcotyledons accurately reflected the damage to the uterus. Plasma progestagen profiles throughout gestation also provided evidence for placental pathology and fetal stress.     

Dynamic obstructions of the equine upper respiratory tract. Part 1: observations during high-speed treadmill endoscopy of 600 Thoroughbred racehorses

REASONS FOR PERFORMING STUDY AND OBJECTIVE: To review the prevalence of single and complex forms of dynamic airway obstructions within a large group of Thoroughbred horses in training referred for investigation of poor performance. METHODS: Video-endoscopic recordings of the upper respiratory tract made during a standardised treadmill exercise test of 600 Thoroughbred racehorses were reviewed and analysed in real time and slow motion to identify dynamic collapse by the tissues bordering onto the pharyngeal and laryngeal airways. RESULTS: Dynamic collapse within the nasopharynx or larynx was confirmed in 471 of the 600 horses. Dorsal displacement of the soft palate (DDSP; 50%) and palatal instability (33%) were the disorders most frequently identified. It was concluded that deglutition is not a significant event in the triggering of DDSP. Complex forms of dynamic collapse were present in 30% of the horses with upper respiratory tract obstructions. A significant influence of age on the prevalence of DDSP and dynamic laryngeal collapse was identified. There was an increased risk of DDSP in younger horses, and of laryngeal collapse in older horses. No association with gender or format of racing was identified. CONCLUSIONS AND POTENTIAL RELEVANCE: Palatal instability and DDSP comprised the most frequently encountered forms of dynamic collapse within the upper respiratory tract of the Thoroughbred racehorses in this study and are probably expressions of the same nasopharyngeal malfunction. Complex obstructions, i.e. where more than one structure collapses into the airway, occur frequently and therefore treatments that address solitary disorders may often be unsuccessful. Younger horses were found to be at greater risk of sustaining DDSP while older horses seemed more at risk to vocal cord collapse but not to collapse of the arytenoid cartilage itself.     

Radiographic changes in Thoroughbred yearlings. Part 1: Prevalence at the time of the yearling sales

REASONS FOR PERFORMING STUDY: Radiography in presale examinations of TB yearlings has become standard practice in recent years. OBJECTIVES: To describe the prevalence and distribution of radiographic changes in the fetlocks, carpi, tarsi, stifles and fore feet of Thoroughbred yearlings in central Kentucky when these joints were examined as part of routine pre- and post sale evaluations. METHODS: Horses subjected to radiographs included the fore (n = 1127) and hind (n = 1102) fetlocks, carpi (n = 1130), tarsi (n = 1101), stifles (n = 660) and fore feet (n = 300). Radiographic changes were categorised by location and type of change present (e.g. lucency, fragment) for each series. RESULTS: In the fore fetlocks 1.6% had fragmentation of the proximal dorsal first phalanx and 0.5% fragmentation of the proximal palmar aspect. In the hind fetlocks 5.9% had fragmentation at the plantar aspect and 3.3% fragmentation dorsally. Lucencies, fragments or loose bodies were detected at the dorsal aspect of the distal third metacarpus in 2.8% and, at the same location on the third metatarsus, 3.2%. Most yearlings (98%) had vascular channels in the proximal sesamoid bones and irregular vascular channels (> 2 mm wide or with nonparallel sides) were more common (79%) than regular vascular channels (56%). The intermediate ridge of the distal tibia was the most common location for fragmentation in the tarsus (4.4%). CONCLUSIONS: While some radiographic changes, e.g. vascular channels in proximal sesamoid bones, are very common in Thoroughbred yearlings, others, e.g. fragmentation or subchondral lucency within joints are quite rare usually affecting less than 5% of the population. POTENTIAL RELEVANCE: Veterinarians should expect to find radiographic changes in Thoroughbreds presented for examination prior to the yearling sales. The rarity of some changes thought to affect soundness or racing performance will make further invesigation of these conditions in horses without clinical signs more difficult.     

Molecular characterisation of 'strangles' outbreaks in the UK: the use of M-protein typing of Streptococcus equi ssp. equi

Reasons for performing study: Strangles is the most commonly diagnosed and important infectious disease of horses worldwide. Very little is known about the temporo‐spatial and molecular epidemiology of strangles. The disease is not notifiable in the UK and there are few published data on the geographical locations of outbreaks. Objective: To investigate whether typing of a surface protein (SeM) of Streptococcus equi ssp. equi (S. equi), the causative agent of strangles, is a useful epidemiological tool. Methods: The variable region of the SeM gene was amplified from 145 isolates of S. equi by PCR and sequenced. Different SeM gene alleles were assigned based on the SeM database, grouped into phylogenetic clusters using split decomposition analysis and plotted against the submitting veterinary practices. Results: In this study 21 S. equi SeM alleles were found, including 9 previously unidentified alleles and representing 4 phylogenetic groups. S. equi containing SeM alleles 9 and 7 were the most commonly isolated and there was a high number of low frequency alleles. The occurrence of an outbreak cluster in the north‐west of the UK is also reported. Conclusions: Strangles outbreaks can be differentiated on the basis of their SeM allele sequences. The data provide further evidence of SeM mutation leading to the emergence of novel, but related SeM alleles that are geographically linked. Sequencing of the SeM gene is a useful tool for the elucidation of strangles epidemiology at a regional and a national level. Potential relevance: This technique may allow differentiation or linkage of strangles outbreaks and as such may be an effective tool for local as well as national and international disease surveillance.     

马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响

旨在研究马链球菌兽疫亚种（Streptococcus equi ssp.zooepidemicus,SEZ）烯醇化酶（enolase,Eno）对小鼠肺泡巨噬细胞（RAW264.7）吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶（rEno）,采用台盼蓝活细胞计数法,判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后,用SEZ作用于细胞并检测细胞吞菌数量,判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术（SILAC）和蛋白质谱分析技术（LC-MS/MS）,筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现,10 μg·mL^-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性,且10 μg·mL^-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用（P<0.01、P<0.05）。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白（dynactin subunit protein 2,Dctn）、整合素α-M蛋白（integrin alpha-M）等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白,发现rEno可减少RAW264.7细胞对SEZ的吞噬,互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。     

Effects of Enolase of Streptococcus equi ssp. zooepidemicus on Phagocytic Functions of Alveolar Macrophages in Mice

To investigate the effect of enolase (Eno) of Streptococcus equi ssp. zooepidemicus (SEZ) on phagocytosis of mouse alveolar macrophages (RAW264.7). Recombinant enolase (rEno) was obtained by constructing prokaryotic expression plasmid, and the cytotoxicity of rEno protein on RAW264.7 cell proliferation was determined by trypan-blue living cell count method. After the rEno protein was incubated with RAW264.7 cells, SEZ was applied to the cells and the quantity of bacteria being phagocytosed was detected to determine the phagocytic activity of RAW264.7 cells. Further, candidate proteins that might interact with SEZ Eno in RAW264.7 cells were screened by live cell stable isotope labeling (SILAC) and protein spectrum analysis (LC-MS/MS). It was found that protein treatment (rEno,10 μg·mL^-1) had significant cytotoxic effects on RAW264.7 cells. Treatment of RAW264.7 cells with 10.0 μg·mL^-1 rEno protein for 2 and 4 hours could significantly inhibit the phagocytosis of RAW264.7 cells (P<0.01, P<0.05). In RAW264.7 cells, dynactin subunit protein 2 (Dctn), integrin alpha-M and about 17 proteins that might interact with Eno were preliminarily identified as rEno interaction proteins. The rEno recombinant expression protein was obtained in this study, and it could reduce the phagocytosis of RAW264.7 cells to SEZ. Preliminary screening of interacting proteins also laid a foundation for further revealing the mechanism of Eno in the anti-phagocytosis of SEZ.     

新疆地区3株驴源马链球菌马亚种新SeM基因型的鉴定与进化分析

从新疆地区某驴养殖场获得了3株驴腺疫链球菌分离株HTP133、HTP123和HTP232.为了解这3株驴腺疫链球菌的生物学特性和确定其分子分型,本研究对其进行了生化特性、药敏特性的检测,对16S rRNA进行序列对比分析,并利用PCR扩增SeM等位基因和测序鉴定其基因型.研究结果表明3株分离菌均为马链球菌马亚种.药敏试...     

致病性马兽疫链球菌的分离鉴定、生物学特性及其M-like基因的序列分析

为鉴定锦州某猪场引起关节炎病疫情的病原,本研究将患猪关节液在血平板培养基上培养进行细菌分离;对分离细菌进行形态学观察、生化反应、致病性试验及药物敏感性试验;并对其M-like基因进行克隆和测序分析.结果在显微镜下观察到周围具有透明的β溶血环的革兰氏阳性中长链球菌,经生化反应及M-like基因鉴定为马兽疫链球菌(命名为10JZ12);分离株10JZ12对小鼠的致死剂量为1.75× 108 cfu;并对苯唑西林、青霉素G、红霉素、克拉霉素、克林霉素、米诺环素、环丙沙星、左氟沙星、多粘菌素B、呋喃妥因敏感,对氯霉素、四环素、诺氟沙星、复方新诺明不敏感.     

马链球菌兽疫亚种PGK蛋白的免疫保护试验

为了阐明马链球菌兽疫亚种(SEZ)PGK蛋白的免疫保护作用,使该菌引起猪、马、牛等多种动物的链球菌病得到有效控制,本试验构建PGK重组表达载体,纯化重组蛋白,对其免疫效力进行系统评价。结果表明:PGK蛋白可以诱导高滴度的血清Ig G抗体,并且可提供一定的免疫保护效力;Real-time PCR技术分析表明,PGK是一个重要的体内诱导抗原;并且可诱导高水平的Th1和Th2型免疫应答,揭示PGK在SEZ治病过程中起到重要作用,为新型疫苗的研制奠定了基础。     

Subscapular lymph node abscessation as a result of metastatic Streptococcus equi subspecies equi infection: An atypical presentation of bastard strangles in a mare

The case reported here represents an atypical presentation of bastard strangles in an 18‐year‐old Arab mare. The horse initially presented for progressive neck pain characterised by reluctance to lateral and ventroflexion of the neck. Subsequent diagnostics revealed a subscapular abscess and aspirates of the mass cultured positive for Streptococcus equi ssp. equi. Surgical drainage and debridement of the abscess was performed under general anaesthesia. Six months post surgery, the mare had made a complete recovery.