Escherichia coli type III secretion system 2: a new kind of T3SS?

Type III secretion systems (T3SSs) are employed by Gram-negative bacteria to deliver effector proteins into the cytoplasm of infected host cells. Enteropathogenic Escherichia coli use a T3SS to deliver effector proteins that result in the creation of the attaching and effacing lesions. The genome sequence of the Escherichia coli pathotype O157:H7 revealed the existence of a gene cluster encoding components of a second type III secretion system, the E. coli type III secretion system 2 (ETT2). Researchers have revealed that, although ETT2 may not be a functional secretion system in most (or all) strains, it still plays an important role in bacterial virulence. This article summarizes current knowledge regarding the E. coli ETT2, including its genetic characteristics, prevalence, function, association with virulence, and prospects for future work.     

Efficacy of a type 2 PRRSV modified live vaccine (PrimePac™ PRRS) against a Thai HP-PRRSV challenge

The Chinese highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) has caused a severe threat to the pig population in Southeast Asian countries. The purpose of this study was to investigate the efficacy of a type 2 PRRSV modified live vaccine (PrimePac? PRRS, lineage 7) against a Thai HP-PRRSV (10PL01, lineage 8). Three-week-old PRRSV-free pigs were randomly assigned into three groups. Vaccinated challenged group (group 1, n?=?16) was immunized with PrimePac? PRRS vaccine at 3 weeks old. The unvaccinated challenged group (group 2, n?=?16) was injected with PBS at 3 weeks old, and unvaccinated unchallenged group (group 3, n?=?10) was served as a negative control. At 9 weeks old, all groups, except the negative control group, were challenged with the Thai HP-PRRSV. All pigs were monitored daily during 10 days post-infection (dpi) and were necropsied at 10 and 17 dpi. The results revealed that vaccinated challenged pigs showed significantly lower (p?<?0.05) mean rectal temperatures, clinical respiratory scores, lung lesion scores, and levels of virus load in serum and lung tissue compared with the unvaccinated challenged pigs. Moreover, vaccinated challenged pigs exhibited PRRSV-specific serum neutralizing antibodies at the end of the experiment. Our findings indicated that the studied type 2 PRRSV vaccine provided partial protection against the Thai HP-PRRSV infection based on the body temperature, levels of viremia, and the severity of lung lesions. These results demonstrated that partial protection of PrimePac? PRRS vaccine might be useful for controlling HP-PRRSV infection in the endemic area.     

Choice feeding of the egg‐type pullet

1. Egg‐type pullets from 43 to 127 d of age were fed either a high‐protein concentrate (493 g/kg), a protein concentrate (323 g/kg) or a grower diet, together with either whole wheat or whole barley. Further birds received a complete diet and had no choice.

2. Eighty‐four point six, 69.5 and 31.4% respectively of the food consumed by the birds which received the high‐protein concentrate, protein concentrate or grower diet with wheat consisted of the whole cereal. The corresponding figures for the birds which received barley were 73.9, 41.3 and 10.9% respectively.

3. There were no significant differences in rate of food intake or growth between choice‐fed birds receiving whole wheat and the complete diet‐fed birds with no choice.     

Porcine circovirus type 2 in muscle and bone marrow is infectious and transmissible to naïve pigs by oral consumption

Pork products are a possible source of introduction of PCV2 isolates into a pig population. However, limited work has been done on the transmission through meat of porcine circovirus type 2 (PCV2), a virus associated with several disease syndromes in pigs. The objectives of this study were to determine if pork products from PCV2-infected pigs contain PCV2 DNA/antigen and to determine if the PCV2 present in the tissues is infectious by performing in vitro and in vivo studies. Skeletal muscle, bone marrow, and lymphoid tissues from pigs experimentally inoculated with PCV2 were collected 14 days post-inoculation (DPI). The tissues were tested for presence of PCV2 DNA by quantitative real-time PCR, for PCV2 antigen by immunohistochemistry (IHC), and for presence of infectious PCV2 by virus isolation and inoculation of PCV2 naïve pigs. Lymphoid tissues contained the highest amount of PCV2 (positive by PCR, IHC, and virus isolation), bone marrow contained a lower amount of PCV2 (positive by PCR and IHC but negative by virus isolation), and skeletal muscle contained the lowest amount of PCV2 (positive by PCR but negative by IHC and virus isolation). Naïve pigs fed for three consecutive days with either skeletal muscle, bone marrow, or lymphoid tissues all became PCV2 viremic as determined by quantitative real-time PCR on serum starting at 7 DPI. The pigs also seroconverted to PCV2 as determined by PCV2 IgM and IgG ELISA. In addition, PCV2 antigen was detected by IHC stains in lymphoid tissues and intestines collected from the majority of these pigs. Results from this study indicate that uncooked PCV2 DNA positive lymphoid tissues, bone marrow, and skeletal muscle from PCV2 viremic pigs contain sufficient amount of infectious PCV2 to infect naïve pigs by the oral route.     

武明欣1，2王际英2李宝山2张德瑞3蔡胜昌1，2王世信2孙永智2张利民2?

本试验旨在研究酶制剂对刺参生长、体成分、免疫能力及氨氮胁迫下免疫酶活力和热休克蛋白70含量的影响。试验共配制粗蛋白质含量为20％的试验饲料5种,首先配制不含酶制剂的基础饲料作为对照,然后分别以0．04％木聚糖酶、0．10％纤维素酶、0．01％淀粉酶和0．15％复合酶（由0．04％木聚糖酶、0．10％纤维素酶和0．01％淀粉酶组成）等量替代基础饲料配方中的α－淀粉。每种饲料设3个重复,每个重复放养体重（4．02±0．04） g 的幼参40头。试验期为56 d。结果表明：1）木聚糖酶、纤维素酶、淀粉酶、复合酶组的增重率分别比对照组提高了17．38％（P＜0．05）、7．63％（P＜0．05）、1．88％（P＞0．05）和20．75％（P＜0．05）,特定生长率分别比对照组提高了0．25％／d（P＜0．05）、0．03％／d（P＜0．05）、0．01％／d（P＞0．05）、0．30％／d（P＜0．05）。2）饲料中添加木聚糖酶显著提高了刺参体壁粗蛋白质的含量（ P＜0．05）,添加纤维素酶、淀粉酶和复合酶显著提高了刺参体壁粗蛋白质、粗脂肪和粗灰分的含量（ P＜0．05）。3）饲料中添加木聚糖酶、纤维素酶和复合酶显著提高了体腔液中过氧化氢酶的活力（ P＜0．05）;添加淀粉酶显著提高了体腔液中溶菌酶、超氧化物歧化酶、过氧化氢酶活力（ P＜0．05）。4）在120 h氨氮胁迫过程中,各组体腔液中溶菌酶、超氧化物歧化酶、过氧化氢酶活力及热休克蛋白70含量基本呈先上升后下降趋势。除纤维素酶组外,其余加酶组体腔液中热休克蛋白70含量的最大值均高于对照组。综上,饲料中适量添加木聚糖酶、纤维素酶、淀粉酶或其复合酶均能显著促进刺参生长,改善体成分,提高免疫能力,木聚糖酶的促生长作用更明显,而淀粉酶的促免疫作用更明显。氨氮胁迫下,饲料中适量添加木聚糖酶、纤维素酶、淀粉酶或其复合酶均能使刺参更迅速和强烈地表现出免疫抵抗反应。     

Astrovirus as a possible cause of congenital tremor type AII in piglets?

Background

Congenital tremor is associated with demyelination of the brain and spinal cord and is clinically noted as outbreaks of trembling and shaking in newborn piglets during a limited time-period. Six forms of the disease have been described, where form AII may be caused by an, as yet, unidentified viral infection. This study aimed to investigate the presence of astrovirus and circovirus by sequencing and polymerase chain reaction (PCR) analysis and by relating the findings to the occurrence of disease and lesions in the brain, in 4–6 days-old piglets obtained from a clinical outbreak of congenital tremor.

Results

In piglets with congenital tremor, there were mild to moderate vacuolar changes of the white matter in the cerebrum, brain stem and cerebellum. In healthy piglets, less conspicuous vacuolar changes were detected. One healthy and one diseased piglet were positive for porcine circovirus type 2. The nested pan-PCR showed the presence of astrovirus in at least one brain region in all piglets and by sequencing, two different porcine astrovirus lineages were identified.

Conclusions

The results do not support previous studies identifying porcine circovirus type 2 as the cause of congenital tremor. The demonstration of astrovirus in the brain of piglets suffering from congenital tremor is interesting. However, astrovirus was demonstrated in both healthy and diseased individuals and therefore, further studies are warranted to determine the possible involvement of astrovirus in the pathogenesis of congenital tremor in pigs.     

Thermoneutral zone and resting metabolic rate of growing white leghorn‐type chickens

1. Metabolic rate was determined once a week in pullets from commercial flocks of the local White Leghorn strain from the age of 1 d to 13 weeks.

2. The equations relating minimal resting metabolic rate and body weight were: for body weights of 40 to 163 g Y ‐ 1–83 W ^0.986, and for 163 to 1 200 g Y = 11.64 W ^0.623, where Y = O₂ consumption (ml/bird h) and W = body weight (g).

3. The relationship between lower critical temperature (T_d) and age may be described by two equations: for pullets of 1 to 21 d of age Y = 35 ‐ 0.286 X, and for 21 to 70 d of age Y = 63.3 X^{‐ 0.254}, where Y = T _d(°C) and X = age (d).

4. T_d related to body weight is also described by two equations: for body weights of 40 to 163 g Y = ’42 ‐ 1.18 W^0.480 , and for 163 to 800 g r = 42 ‐ 3.60 W^0.260

5. These equations could be used to calculate the T_d on a day‐to‐day basis related to age or body weight.     

致病性鸡大肠杆菌type1菌毛fimC基因的克隆与序列测定

根据国外发表的人致病性大肠杆菌fimC基因序列,在其保守区设计了带有BamHⅠ/HindⅢ酶切位点的一对引物,应用PCR技术以鸡致病性大肠杆菌O2基因组DNA为模板扩增到一个片段,大小约为700bp,将扩增产物克隆到pMD18-T载体上,并转化于大肠杆菌TG1宿主菌,经酶切和筛选,得到阳性重组质粒.通过对阳性重组质粒核酸序列测定,确定此DNA片段为鸡大肠杆菌fimC基因.     

Divergent selection for delayed‐type wattle reaction of domestic fowls to BCG antigen

1. Two‐way selection for delayed‐type hypersensitivity wattle reaction (DWR) competence to BCG antigen was carried out over 4 generations, using a flock of White Leghorn chickens. DWR was measured by intradermal injection of BCG into the wattle of chickens.

2. Selection for DWR was effective. The generation means of average selection differential, selection response and realized heritability were 1–15 mm, 0–77 mm and 0–70 respectively. Correlated responses were found in body weight, egg weight and hatchability. In all of these traits, the L line (selected for low wattle reaction) showed higher values than the H line (high reaction).

3. Specific trends in gene frequencies were observed at the major histocompatibility B and alkaline phosphatase Akp loci.

4. Significant line differences were found in graft‐versus‐host reaction competence and Marek's disease resistance. In both characters, the L line was higher than the H line. No line differences were found in phytohaemagglutinin reponse, phagocytic activity or immune responses.     

2株猪圆环病毒2型ORF2的克隆及序列分析

根据GenBank中猪圆环病毒2型(PCV-2)的核酸序列,设计一对引物,采用PCR方法从疑似断奶仔猪多系统衰竭综合征(PMWS)的死亡仔猪组织病料中扩增出ORF2基因,将其克隆到pMD18-T载体上,筛选获得重组质粒pMD18T-ORF2。序列分析表明,克隆的ORF2基因与其他PCV-2ORF2的分离株的同源性在91.5%～99.4%,与杭州分离株的同源性达到99.4%,表明我国各地区之间PCV-2的分离株存在较大的差异。     

猪IL2基因重组对PPV VP2-PCV2ORF2真核质粒免疫原性增强的研究

将猪IL2成熟肽基因、PPV VP2基因和PCV2 ORF2抗原多肤基因定向克隆至真核表达载体pCI-neo,构建重组质粒pCI-ORF2-VP2和pCI-IL2-ORV2-VP2,用脂质体介导法将其转染PK15细胞,采用RT-PCR法和间接免疫荧光法检测重组质粒在体外的表达情况.并以小鼠为动物模型,分别将pCI-IL2-ORF2-VP2质粒、pCI-ORF2-VP2质粒、pCI空载体、猪细小灭活苗和圆环亚单位苗通过肌注进行免疫,检测免疫小鼠脾脏淋巴细胞转化功能,外周血T淋巴细胞亚群的动态变化及血清抗体效价.结果表明,重组质粒在体外能正常表达,pCI-IL2-ORF2-VP2免疫小鼠脾脏淋巴细胞转化功能,CD4+、CD8+T淋巴细胞的数量和血清PPV/PCV2抗体的OD值在二免后均高于或显著高于pCi-ORF2-VP2对照组.结果表明,猪IL2基因重组能显著增强VP2/ORF2核酸疫苗诱导的免疫应答.     

The association of Clostridium botulinum type C with equine grass sickness: a toxicoinfection?

The cause of grass sickness, an equine dysautonomia, is unknown. The disease usually results in death. Gastrointestinal (GI) dysfunction is a common clinical manifestation in all forms of the disease. It is generally thought that equine grass sickness (EGS) is caused by an ingested or enterically produced neurotoxin which is absorbed through the GI tract. Clostridium botulinum was first implicated as a causative agent when it was isolated from the GI tract of a horse with EGS in 1919. The aim of the present study was to investigate the hypothesis that EGS results from toxicoinfection with C. botulinum type C: growth of the bacterium in the GI tract with production of toxin (BoNT/C). Ileum contents and faeces from horses with EGS were investigated for BoNT/C, and indirectly for the presence of C. botulinum type C, and compared with control samples from horses without EGS. BoNT/C was detected directly by ELISA in the ileum of 45% (13/29) of horses with EGS compared to 4% (1/28) of controls, and in the faeces of 44% (20/45) of horses with EGS compared to 4% (3/77) of controls. Levels of up to 10 Mlg toxin/g wet weight of gut contents were observed. The one control horse with detectable toxin in the ileum had been clinically diagnosed as having acute EGS, but this was not confirmed by histopathology. The organism was detected indirectly by assaying for BoNT/C by ELISA after enrichment in culture medium. C. botulinum type C was shown to be present in 48% (14/29) of ileum samples and 44% (20/45) of faecal samples from horses with EGS, compared with 7% (2/27) of ileum samples and 8% (6/72) of faecal samples from controls. These results support the hypothesis that EGS results from a C. botulinum type C toxicoinfection.     

H2O2与Fe2+、Cu2+、Zn2+协同作用对桑自由基伤害和保护酶活性的影响

研究了H2O2与Fe2+等金属离子产生的自由基胁迫对桑树生理特性的影响,旨在为桑树的抗逆栽培提供理论参考。以桑树叶片为材料,研究H2O2与Fe2+、Cu2+、Zn2+协同作用对桑自由基伤害和保护酶活性的影响。结果表明:经H2O2-Fe2+、H2O2-Cu2+和H2O2-Zn2+3种体系溶液处理的桑.OH含量分别提高34.38%、8.14%和5.43%;O2.-含量分别降低78.77%、54.75%和63.84%;1,1-二苯基苦基苯肼(DPPH)清除率分别降低44.60%、57.34%和54.64%;超氧化物歧化酶(SOD)活性分别提高44.45%、36.02%和28.28%;多酚氧化酶(PPO)活性分别降低68.18%、86.58%和54.78%;过氧化氢酶(CAT)活性分别降低97.46%、96.57%和68.02%;过氧化物酶(POD)活性分别降低22.22%、提高7.42倍和66.67%。H2O2与Fe2+等的协同作用破坏了桑细胞内自由基动态平衡,导致自由基含量提高,保护酶活性受到显著影响。     

猪场圆环病毒2型(PCV-2)

葡萄球菌病主要是由金黄色葡萄球菌引起的家禽的一种急性或慢性传染病,在临床上常表现多种类型,如关节炎、腱鞘炎、脚垫肿、脐炎和葡萄球菌性败血症等。