炭疽杆菌双毒力因子荧光PCR检测方法的建立及应用

由炭疽杆菌（Bacillus anthracis）引起的炭疽（anthrax）是严重危害人类和家畜健康的重大传染病。为了提高出入境口岸对不明粉未快件样品的查验效率,防范通过包裹夹带生化武器,本研究使用Taq Man探针技术,建立了针对炭疽杆菌两种毒力质粒pOX1和pOX2的双重荧光PCR检测方法,并对该方法的特异性、灵敏性和稳定性进行了试验。结果显示：本研究建立的炭疽杆菌双重荧光PCR检测方法具有较强的特异性,只特异性检出炭疽杆菌pOX1和pOX2,而与金黄色葡萄球菌等19种对照病原菌均无交叉反应;灵敏度与单重荧光PCR方法相近,最低检测限可达10 copies/μL;稳定性好,反应体系低温保存2、4、6、8个月后,检测阳性质粒的Ct值和峰值相差不大。应用该方法检测600份猪牛羊组织样本,均未检出炭疽杆菌阳性核酸。本研究建立的双重荧光PCR检测方法为炭疽杆菌的快速检测提供了技术支撑。     

Effectiveness of calcium hypochlorite,quaternary ammonium compounds,and sodium hypochlorite in eliminating vegetative cells and spores of Bacillus anthracis surrogate

BackgroundThe spore-forming bacterium Bacillus anthracis causes anthrax, an often-fatal infection in animals. Therefore, a rapid and reliable strategy to decontaminate areas, humans, and livestock from B. anthracis is very critical.ObjectivesThe aim of this study was performed to evaluate the efficacy of sodium hypochlorite, calcium hypochlorite, and quaternary ammonium compound (QAC) sanitizers, which are commonly used in the food industry, to inhibit spores and vegetative cells of B. anthracis surrogate.MethodsWe evaluated the efficacy of sodium hypochlorite, calcium hypochlorite, and a QAC in inhibiting vegetative cells and spores of a B. anthracis surrogate. We treated a 0.1-mL vegetative cell culture or spore solution with 10 mL sanitizer. The samples were serially diluted and cultured.ResultsWe found that 50 ppm sodium hypochlorite (pH 7), 1 ppm calcium hypochlorite, and 1 ppm QAC completely eliminated the cells in vegetative state. Exposure to 3,000 ppm sodium hypochlorite (pH 7) and 300 ppm calcium hypochlorite significantly eliminated the bacterial spores; however, 50,000 ppm QAC could not eliminate all spores.ConclusionsCalcium hypochlorite and QAC showed better performance than sodium hypochlorite in completely eliminating vegetative cells of B. anthracis surrogate. QAC was ineffective against spores of the B. anthracis surrogate. Among the three commercial disinfectants tested, calcium hypochlorite most effectively eliminated both B. anthracis vegetative cells and spores.     

地衣芽孢杆菌与其他微生物产酶能力的比较

试验对能在动物肠道产酶的芽孢产品—“益畜宝”的有效成分—地衣芽孢杆菌(Bacillus licheniformis)的产酶特性进行了初步定性研究。在单一碳源培养基上的观察结果表明，该地衣芽孢杆茵能产生植酸酶、淀粉酶及蛋白酶。相同的试验条件下，大肠杆菌和啤酒酵母菌都不产生植酸酶。实验结果还表明，大肠杆菌虽然能产生淀粉酶和蛋白酶，但其活性比地衣芽孢杆菌要低得多，而啤酒酵母菌则不产生淀粉酶和蛋白酶。     

Fluorescence serologic and histologic studies of antigen distribution in parvovirus infections in dogs and cats]

In organs known as replication sites of parvovirus and additionally in stratified squamous epithelia from different regions of 35 dogs with lethal parvovirus enteritis and 19 cats with lethal panleukopenia the distribution of antigen and the prevalence of histologic lesions were studied by immunofluorescence and histologic techniques. The findings showed no striking differences between dogs and cats. Antigen was found in the epithelia of the dorsal side of the tongue (96.3%), pharynx (81.0%), esophagus (50.0%), ventral side of the tongue (20.4%) and planum nasale (5.6%). The epithelia of the skin and the mucous membranes of the male and female genital organs never contained antigen. Moreover antigen was detected in the mucosa of the small intestine (85.2%), bone marrow (81.6%), spleen (79.6%), thymus (66.7%), mesenteric lymph nodes (60.4%), tonsilla palatina (58.5%), smooth muscle cells of the tunica muscularis of the small intestine (9.3%) and myocardium (1.9%). The histologic lesions found in intestinal mucosa, lymphatic tissues, bone marrow and myocardium are in accordance with literature. The tunica muscularis of the small intestine and the epithelium of tongue, pharynx and esophagus contained intranuclear inclusion bodies, the epithelium of tongue and pharynx showed necroses additionally. Tongue- and esophagus-erosions as well as -ulcers reported mainly in panleukopenia according to our results may be explained as a sequel of parvovirus infection of the epithelia of these organs. The prevalence of antigen and intranuclear inclusion bodies in the myocardium and smooth muscle cells of the small intestine--tissues with low or no mitotic activity,--allows to question the postulate of the parvoviruses' exclusive affinity to rapidly dividing cells.     

复合菌剂及酸化剂对玉米秸秆青贮有氧暴露后的影响

为探讨复合菌剂和酸化剂对玉米秸秆青贮有氧稳定性及有氧暴露期间微生物数量、pH值、粗蛋白、干物质量的影响,以玉米秸秆为青贮原料,设置3个处理:对照组、复合菌剂组、复合菌剂+酸化剂组.青贮90 d.有氧暴露1、3、7、11 d分别取样测定微生物数量、pH值、粗蛋白、干物质量.结果显示,与对照组相比,复合菌剂组、复合菌剂+酸...     

优良抑菌活性乳酸菌对玉米青贮及有氧暴露期微生物数量和pH的影响

为了探讨乳酸菌对全株玉米青贮及有氧暴露后青贮饲料中乳酸菌、好氧细菌、酵母菌和霉菌数量及其pH的影响,进一步筛选出可提高青贮饲料品质和有氧稳定性的乳酸菌接种剂,将实验室前期从甘肃各地玉米秸秆青贮饲料中分离筛选获得的5株产酸快、多且具有抑菌活性的优良乳酸菌分别添加全株玉米进行青贮,分析青贮过程和有氧暴露后青贮饲料中乳酸菌、好氧细菌、酵母菌和霉菌数量的动态变化及pH。结果显示,在青贮过程和有氧暴露后,分别添加肠膜明串珠菌肠膜亚种B1-7、戊糖片球菌B2-3、植物乳杆菌B3-1、屎肠球菌B5-2和发酵乳杆菌E2-3的各处理组乳酸菌总数均显著高于对照组,而好氧细菌、酵母菌和霉菌数量均显著低于对照组,pH亦低于对照组。其中B1-7和B5-2处理组在青贮初期乳酸菌总数最多,从青贮第7天开始到有氧暴露的30 d内,始终是B3-1处理组乳酸菌总数最多,好氧细菌、酵母菌和霉菌数量最少、pH最低。以上结果表明这5株乳酸菌具有提高青贮饲料品质和有氧稳定性的潜力,其中植物乳杆菌B3-1的效果最好。     

Study of the role of Chlamydia,Mycoplasma, Ureaplasma and other microaerophilic and aerobic bacteria in uterine infections of mares with reproductive disorders

In six healthy mares and 24 mares showing reproductive disorders swab samples were taken from the fossa clitoridis to isolate Taylorella equigenitalis, and from the uterus to isolate mycoplasmas, ureaplasmas and other aerobic bacteria. Swab samples were also taken from the uterus for Chlamydia antigen ELISA and Chlamydia PCR studies. The uterus of 27 mares was examined cytologically, and biopsy samples were taken from the endometrium for histological examinations and for immunohistochemical examinations aimed at the detection of chlamydiae. T. equigenitalis, mycoplasmas, ureaplasmas and chlamydiae could not be detected from any of the mares examined. Aerobic facultative pathogenic bacteria were isolated from mares with endometritis in four cases. In 18 out of 22 mares with endometritis (82%) no infective agents could be demonstrated. Further studies are needed to elucidate the relative importance of non-infectious causes of endometritis and of anaerobic bacteria often detectable in the uterus in the aetiology of the reproductive disorders observed.     

Effects of Bacillus subtilis B10 spores on viability and biological functions of murine macrophages

This study was conducted to investigate the effects of Bacillus subtilis B10 spores on the viability and biological functions of murine macrophage. RAW 264.7 cells were stimulated both with and without B. subtilis B10 spores for 12 h. Then cell viability was determined to evaluate the cytotoxic effect of B. subtilis B10 spores to the cells, and the activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH), the production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), and the secretion of inflammatory cytokines were measured to analyze the functions of macrophages. The results showed that B. subtilis B10 spores were not harmful to RAW 264.7 cells and they also strongly enhanced the activities of ACP and LDH (P < 0.01), remarkably increased NO and iNOS production (P < 0.01), and significantly stimulated the secretion of pro‐inflammatory cytokines, including tumor necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin‐1 beta (IL‐1β), IL‐6, IL‐8 and IL‐12 (P < 0.01) while they reduced anti‐inflammatory cytokine IL‐10 (P < 0.01). The outcomes suggest that B. subtilis B10 spores are not only safe for murine macrophages, but also can activate these cells and enhance their immune function. The above findings suggest that B. subtilis B10 spores are potentially probiotic.     

Survival of bacteria on zinc-coated and aluminum materials]

The survival of St. aureus. E. coli, and of salmonellae was tested without any protein error and with it on new galvanized and with disinfection disturbed plate and compared with the survival of these microorganisms on new aluminous plate which was considered to be inert. The investigation by means of the impression method lasted for seven days. The growth of the microorganisms was evaluated according to the index of their survival. A distinct oligodynamic effect of the galvanized plate was proved especially on gram-negative bacteria. The effectiveness increased by a disturbing of these plates with a long-term disinfection in a concentrated solution of chloramine BS. An antimicrobial effect was recorded also in aluminous plates disturbed by disinfection. On both kinds of plate the effectiveness was lowered by a protein error--by an adding of blood serum to the broth suspensions.     

Use of a foaming disinfectant and cleaner to reduce aerobic bacteria on poultry transport coops

Poultry transportation coops are rarely washed and have been demonstrated to be a point of cross-contamination of broiler carcasses. Foaming disinfectants and cleaners, commonly used within processing plants, may be used to clean and disinfect poultry transportation coops. In this study, homogenized fecal material was evenly applied to the floors of precleaned broiler transportation coops and allowed to dry. Treatments consisted of a water rinse, a foam additive alone, foaming cleaner, and a disinfectant plus a foam additive. All foaming treatments were applied using a compressed air foam system (2,271 L/min; 600 gal/min), similar to what firefighters would use. A high-pressure water rinse (HPWR) was added prior to or following the treatments to determine whether rinsing prior to product application or rinsing after product application would improve efficacy. Based on our data, a compressed air foam system may be used in combination with a commercially available disinfectant or foam cleaner to reduce aerobic bacteria on the surfaces of commercial broiler transportation coops. Furthermore, the addition of a HPWR did not further reduce the level of aerobic bacteria on broiler transportation coop floors.     

Field evaluation of the efficacy of a probiotic containing Bacillus licheniformis and Bacillus subtilis spores, on the health status and performance of sows and their litters

The aim of this study was to assess the efficacy of BioPlus 2B, a probiotic containing Bacillus licheniformis and Bacillus subtilis spores, on the health status and productivity of sows and their litters. A total of 109 gilts and sows were allocated into two experimental groups, as follows: untreated controls (UC) and BioPlus 2B (same feeding as the UC group plus BioPlus 2B) at a dose of 400 g/ton of feed (equal to 1.28 x 10(6) viable spores/g of feed). Treatment started from the day of allocation (14 days prior to the expected farrowing) up to the weaning day. Homogeneity of the groups was satisfied with regard to the parity. From the results it was evident that BioPlus 2B supplementation of the feed improved gilt/sow performance as shown by: (i) the increase of sow feed consumption during the first 14 days postpartum and (ii) the decrease of sow weight loss during the suckling period. Certain blood and milk parameters were significantly improved, as shown by higher serum cholesterol and total lipids concentrations and higher milk fat and protein content at mid-suckling period. As a consequence, a positive effect was also noticed as regard litter health and performance characteristics in terms of: (i) decrease in piglet diarrhoea score, (ii) decrease in pre-weaning mortality thus leading to increase in the number of weaned piglets per litter and (iii) increase in piglet body weight at weaning. Moreover, BioPlus 2B tended to improve the health status and fertility of sows demonstrating: (i) tendency to a lower proportion of sows with Mastitis-Metritus-Agalactia (MMA) problems and (ii) lower proportion of sows returning to oestrus.     

Influence of genotype and diet on steer performance, manure odor, and carriage of pathogenic and other fecal bacteria. II. Pathogenic and other fecal bacteria

This study assessed the influence of cattle genotype and diet on the carriage and shedding of zoonotic bacterial pathogens and levels of generic Escherichia coli in feces and ruminal contents of beef cattle during the growing and finishing periods. Fifty-one steers of varying proportions of Brahman and MARC III [0 (15), 1/4 (20), 1/2 (7), and 3/4 Brahman (9)] genotypes were divided among 8 pens, such that each breed type was represented in each pen. Four pens each were assigned to 1 of 2 diets [100% chopped bromegrass hay or a diet composed primarily of corn silage (87%)] that were individually fed for a 119-d growing period, at which time the steers were switched to the same high-concentrate, corn-based finishing diet and fed to a target weight of 560 kg. Feces or ruminal fluid were collected and analyzed at alternating intervals of 14 d or less. Generic E. coli concentrations in feces or ruminal fluid did not differ (P > 0.10) by genotype or by growing diet in the growing or finishing periods. However, the concentrations in both feces and ruminal fluid increased in all cattle when switched to the same high-corn diet in the finishing period. There was no effect (P > 0.25) of diet or genotype during either period on E. coli O157 shedding in feces. Forty-one percent of the steers were positive for Campylobacter spp. at least once during the study, and repeated isolations of Campylobacter spp. from the same steer were common. These repeated isolations from the same animals may be responsible for the apparent diet (P = 0.05) and genotype effects (P = 0.02) on Campylobacter in feces in the finishing period. Cells bearing stx genes were detected frequently in both feces (22.5%) and ruminal fluid (19.6%). The number of stx-positive fecal samples was greater (P < 0.05) for 1/2 Brahman steers (42.9%) than for 1/4 Brahman (25.0%) or 3/4 Brahman steers (22.2%), but were not different compared with MARC III steers (38.3%). The greater feed consumption of 1/2 Brahman and MARC III steers may have resulted in greater starch passage into the colon, accompanied by an increase in fecal bacterial populations, which may have further improved the ability to detect stx genes in these cattle. There was no correlation between either ADG or daily DMI and the number of positive samples of E. coli O157, Campylobacter spp., or stx genes, which agrees with our current understanding that these microorganisms occur commonly in, and with no measurable detriment to, healthy cattle.     

温度和乳酸菌添加剂对燕麦青贮发酵品质及氧稳定性的影响

以乳熟期刈割的燕麦为材料,研究温度和乳酸菌添加剂对其青贮发酵品质及有氧稳定性的影响,设置无添加(对照,CK)、商业青贮添加剂Synlac I(SLI处理)和人工筛选菌株Lactobacillus rhamno-sus HT1(HT1处理)3个处理,分别置于25℃、35℃和45℃下青贮60 d后,取样测定青贮燕麦的发酵品...