Evaluation of the stable fly (Stomoxys calcitrans) as a vector of enzootic bovine leukosis

Experiments reported here were directed at 2 questions: (1) Can the stable fly (Stomoxys calcitrans) transmit enzootic bovine leukosis? (2) Could early viremia augment the probability of transmission by this insect? In one vector experiment, calves and bovine leukemia virus (BLV)-infected cows were housed with and without stable flies. The calves were monitored serologically during a 3-month postexposure period, using the agar gel immunodiffusion test. All fly-infested and fly-free calves remained BLV-seronegative. For a second vector experiment, donor calves, newly injected with blood from BLV-infected cows with high virus expression, were tethered alternately between uninoculated, weaned BLV-seronegative calves. These groups were housed with or without flies in 2 replicate trials. The inoculated calves from the first replicate seroconverted at 16 and 23 days after inoculation; the inoculated calves from the second replicate seroconverted at 11, 16, 16, and 37 days after inoculation. All uninoculated calves remained BLV-seronegative. In a manual transmission experiment, 50 unfed stable flies were allowed to complete a meal on each of 3 BLV-seronegative calves after feeding on a BLV-seropositive cow with high (42%) virus expression. One control calf was injected with blood from the cow. Seroconversion occurred in the control calf and 1 calf on which flies were given access. A scanning electron microscopic study was made of the everted and closed mouth parts of the stable fly. Given the lymphocyte count in blood from the cow used in the manual vector transmission experiment, it was calculated that 3,950 mouth part volumes would be necessary to transmit BLV.(ABSTRACT TRUNCATED AT 250 WORDS)     

Some effects of temperature on the adults, eggs and pupae of Stomoxys calcitrans Linnaeus (Diptera: Muscidae)

Adults could only live and reproduce to their full capacity at temperatures between 20 degrees C and 30 degrees C. At 15 degrees C the females laid no eggs, the adult life span was relatively short and the reproductive capacity of females kept at 35 degrees C was low. The thermal histories of the flies had no apparent effect on their later reactions to temperature in any of the parameters tested. The viability rates of S. calcitrans eggs exposed to temperatures between 10 degrees C and 40 degrees C exceeded 84%, but 45 degrees C was lethal. The optimum temperatures for incubation of the eggs was 30 degrees C. Pupae of S. calcitrans seemed to tolerate temperatures between 20 degrees C and 30 degrees C, but their mortalities increased markedly outside this temperature range. Tests showed that pupal mortalities increased linearly with increasing periods of exposure to a temperature of 15 degrees C.     

Acetylcholinesterase of Stomoxys calcitrans (L.) (Diptera: Muscidae): cDNA sequence, baculovirus expression, and biochemical properties

A 2193-nucleotide cDNA encoding acetylcholinesterase (AChE) of the stable fly, Stomoxys calcitrans (L.) was sequenced and expressed in the baculovirus system. The open reading frame encoded a 91 amino acid secretion signal peptide and a 613 amino acid mature protein with 96% and 94% identity to the AChEs of Haematobia irritans (L.) and Musca domestica (L.), respectively. Structural characteristics of M. domestica, H. irritans, and Drosophila melanogaster AChEs were conserved in the S. calcitrans AChE. The recombinant enzyme was inhibited by eserine, coroxon, and paraoxon and exhibited K_m values of 63.9 μM for acetylthiocholine and 96.7 μM for butyrylthiocholine, confirming its biochemical identity as an acetylcholinesterase (EC 3.1.1.7). These data will enable rapid identification and assay for mutations that reduce AChE sensitivity to organophosphate (OP) pesticides, potentially aiding resistance management efforts to prevent fixation of the mutations in pest populations.     

Distribution, seasonality and relative abundance of Stomoxys calcitrans (stablefly) (Diptera: Muscidae) in New Zealand

AIMS: To determine the current distribution, seasonality and relative abundance of Stomoxys calcitrans in New Zealand in order to provide information that could be used to assess risks of transmission of equine infectious anaemia (EIA). METHODS: Adhesive yellow traps were distributed to schools throughout New Zealand and used to detect the presence of S. calcitrans between November 1999 and April 2000 at sites considered likely to be a focus for S. calcitrans breeding and activity. In addition, researchers undertook monthly trapping at six other sites between August 1999 and June 2000 to measure the duration and seasonal periodicity of S. calcitrans flight activity. Veterinary practices and farmers were also surveyed to provide anecdotal evidence of the presence or absence of S. calcitrans, particularly in areas where no flies were trapped by schools. RESULTS: Stomoxys calcitrans was found to occur in both North and South Islands, but principally in locations where dairy farming occurred. The fly was active during most months of the year, except perhaps during July and August, and was particularly active in warmer North Island districts such as the Waikato. Peak activity was recorded from January to May. The fly was more abundant in Northland, Auckland, Waikato, the Marlborough Sounds and Nelson than in other districts, as determined by the number of occasions flies were caught relative to the number of traps set. There are only a few areas, such as around Taupo, the Otago Lakes, Central Otago and the Mackenzie Basin, where S. calcitrans was not trapped or, on the basis of anecdotal evidence, thought not to occur. CONCLUSIONS: Given that other potential vectors of EIA are absent from New Zealand, Taupo, the Otago Lakes, Central Otago and the Mackenzie Basin districts could be considered areas of low risk for EIA transmission to horses. In addition, teachers and school children were capable, in most instances, of supporting a nationwide survey providing the methods were simple and the aims few.     

PRRSV变异株活疫苗和灭活疫苗免疫特性的研究

为进一步研究猪繁殖与呼吸综合征病毒(PRRSV)变异株疫苗的免疫机理和明确PRRSV变异株活疫苗和灭活疫苗各自的免疫特性,本实验分别采用PRRSV变异株(HuN4)活疫苗和PRRSV变异株(JXA1)灭活苗免疫PRRSV抗原和抗体阴性的健康断奶仔猪,免疫后21d用PRRSV变异株HuN4强毒攻毒,ELISA方法检测血清中PRRSV特异的抗体水平及TNF-α、IFN-α、IL-1、IL-6和CRP细胞因子水平,荧光定量RT-PCR方法检测病毒血症的发生和持续情况,并取主要器官进行病理组织学观察。结果表明:HuN4活疫苗组免疫后14d便可检测到PRRSV特异性抗体,攻毒后5d各细胞因子水平升高,攻毒后21d病毒血症完全消失,免疫后各器官没有明显病理变化,攻毒后临床症状和各组织器官病理变化轻微;JXA1株灭活苗组免疫期间没有检测到PRRSV特异性抗体,攻毒后5d～9d各细胞因子水平升高,攻毒后病毒血症持续存在,免疫后各器官有轻微病理变化,攻毒后临床症状和各组织器官病理变化比HuN4活疫苗组严重,但比对照组明显减轻。本实验表明,HuN4活疫苗能够快速有效的激发机体的体液免疫反应,在抵抗PRRSV变异株HuN4强毒攻毒时,临床症状明显优于JXA1灭活苗。     

Immunological responses of fluke-infected and fluke-free cattle to Salmonella dublin and other antigens.

Immune responses to heat-killed Brucella abortus strain 19 and to ovalbumin were compared in 15 fluke-infected and 15 fluke-free Friesian heifers. B abortus was injected 16 weeks and ovalbumin 19 weeks after the oral administration of 1000 metacercariae of Fasciola hepatica. Agglutinating antibody responses to B abortus were similar in both groups. Immediate type hypersensitivity to ovalbumin was apparently suppressed in fluke-infected animals when assessed by active and passive cutaneous anaphylaxis two weeks after sensitisation. However, when assessed by Schultz-Dale responses of intestine, in vitro, 36 weeks after sensitisation there was no difference between the groups. The heifers were subsequently given live Salmonella dublin intravenously. The fluke-infected animals which became carriers of S dublin had the most persistently elevated titres of agglutinating antibodies in their sera and the highest incidence of immediate-type hypersensitivity, as assessed by Schultz-Dale responses of intestine, but the weakest cutaneous delayed hypersensitivity reactions to S dublin. The latter might have been related to lymphopenia which developed after fluke infection. The increased susceptibility of fluke-infected cattle to S dublin cannot be attributed to impaired agglutinin responses but may result from effects on cell-mediated mechanisms.     

Immunological characterization of protective antigens prepared by alkaline treatment of whole cells and from the culture filtrate of Erysipelothrix rhusiopathiae.

Culture filtrate and alkaline-extracted antigens from whole cells of an attenuated strain of Erysipelothrix rhusiopathiae (strain Koganei: serovar 1a) were fractionated with ammonium sulfate; both induced protective immunity in mice. Sephadex G-200 gel filtration revealed three protein fractions in the alkaline-extracted antigen and four protein fractions in the culture filtrate antigen. A fraction in the alkaline extract (NaOH P-2) and in the culture filtrate (CF P-2) induced protection in mice against challenge with a different serovar strain (strain Agata: serovar 5). Anti-NaOH P-2 and anti-CF P-2 mouse sera were protective against different serovars. Glycoprotein fraction derived from CF P-2 antigen by affinity chromatography with Con A-Sepharose 4B did not show protective activity. Western blotting between the antisera (anti-NaOH P-2, Anti-CF P-2 and anti-Koganei strain) and the antigens (NaOH P-2, and sonicated antigens of Agata, Fujisawa and Koganei strains) showed strong recognition of the same bands at 62, 42 and 41 kDa.     

泰乐菌素菌渣酶制剂对肉鸡饲喂效果的研究

试验选取1日龄健康AA商品代肉鸡180羽,随机分成2组,每组3个重复,每个重复30只鸡分别饲喂含0 g/kg(对照组)、3 g/kg(酶制剂组)泰乐菌素菌渣酶制剂的玉米-豆粕型日粮42 d,研究该酶制剂对肉鸡的饲喂效果。结果表明:酶制剂组肉鸡体重和饲料利用效率均显著优于对照组(P<0.05)。酶制剂能使饲料中干物质、粗蛋白质、粗脂肪、粗纤维和粗灰分的表观消化率显著提高(P<0.05),粪中大肠杆菌数和空肠内容物黏度显著降低(P<0.05)。添加酶制剂组肉鸡腹肌、肝脏、肾脏中均未检测到残留的泰乐菌素。     

Solubilization studies on the epicuticular antigens of Strongyloides ratti

Antigens on the epicuticular surface of Strongyloides ratti infective third-stage larvae (L3) were demonstrated by both ferritin-conjugated antibody and indirect fluorescent antibody techniques. The rat antibodies from immune serum that bind to these antigens were chiefly of the IgG2a subclass. Solubilization of these antigens by extraction with detergents, hypertonic salt, organic solvents and by freezing and thawing was limited as measured by the reduction in antibody binding to the epicuticle. The epicuticular antigens were resistant in situ to degradation by a variety of proteases, carbohydrases and lipase. Infectivity of the L3 in the rat was not reduced by prior sensitization with rat antibody. The epicuticular antigens are not completely species-specific since antibody from S. ransomi-infected pigs cross-reacted well with S. ratti L3 antibody. However, high levels of resistance to S. ratti could be induced in rats only by multiple inoculations of heat-killed S. ratti L3.