两种侵袭宿主方式的优势种马胃蝇飞行能力研究

为了解两种不同侵染宿主方式马胃蝇的飞行行为,本研究利用飞行磨系统测定了黑腹胃蝇Gasterophilus pecorum（以牧草为产卵载体）和肠胃蝇G. intestinalis（以宿主体毛为产卵载体）的飞行能力。结果表明:（1）肠胃蝇总飞行时间和距离均显著高于黑腹胃蝇,分别为后者的5.52倍和7.65倍,但平均飞行速度无显著性差异（P>0.05）。（2）黑腹胃蝇雌虫的飞行时间、距离和速度均略高于雄虫,而肠胃蝇雌虫除平均飞行速度外的飞行参数均低于雄虫。（3）肠胃蝇吊飞期间的体重消耗（24.38%）显著高于黑腹胃蝇（14.07%）;黑腹胃蝇雌雄成虫甘油三酯含量均显著下降,但两者差异不显著（P>0.05）。飞行距离差异反映了两种不同侵染宿主方式马胃蝇的飞行能力发生了适应性的变化,而总飞行时间为两种马胃蝇飞行距离差异的主导因素。     

Light and electron microscopy studies of the midgut and salivary glands of second and third instars of the horse stomach bot,Gasterophilus intestinalis

A morphological study of the midgut and salivary glands of second and third instars of Gasterophilus intestinalis (De Geer) (Diptera: Oestridae) was conducted by light, scanning and transmission electron microscopy. The midgut is anteriorly delimited by a proventriculus, without caeca, and is composed of posterior foregut and anterior midgut tissue from which a double‐layered peritrophic matrix is produced. The midgut can be divided into anterior, median and posterior regions on the basis of the structural and physiological variations of the columnar cells which occur along its length. Two other types of cell were identified: regenerative cells scattered throughout the columnar cells, and, more rarely, endocrine cells of two structural types (closed and open). Different secretion mechanisms (merocrine, apocrine and microapocrine) occur along the midgut epithelium. Abundant microorganisms are observed in the endoperitrophic space of the anterior midgut. The origin and nature of these microorganisms remain unknown. No structural differences are observed between the second and third instar midguts. The salivary glands of G. intestinalis second and third instars consist of a pair of elongated tubular structures connected to efferent ducts which unite to form a single deferent duct linked dorsally to the pharynx. Several intermediate cells, without cuticle, make the junction with the salivary gland epithelium layer. Cytological characteristics of the gland epithelial cells demonstrate high cellular activity and some structural variations are noticed between the two larval stages.     

Epizootic abortion related to infections by Chlamydophila abortus and Chlamydophila pecorum in water buffalo (Bubalus bubalis)

Water buffalo (Bubalus bubalis) are affected by high rates of embryonic mortality and abortion related to infectious diseases and non-infectious factors. A number of viral and bacterial infections have been associated with reproductive failure, but there is limited information on the role of chlamydial infections.

In order to investigate the presence and the role of Chlamydiaceae in water buffalo a retrospective study was performed in a herd with a history of reproductive failure. During an 11-month period, the pregnant heifers suffered an abortion rate of 36.8% between the 3rd and 7th month of pregnancy. Antibodies to Chlamydiaceae were detected in 57% of the aborted cows, and in 0% of the overtly healthy cows used as control. By a nested-PCR assay, three of 14 vaginal swabs from aborted animals tested positive for Chlamydophila agents and, additionally, three out of seven aborted fetuses tested positive for Chlamydophila spp., with two being co-infections by Cp. abortus and Cp. pecorum and one being characterised as Cp. abortus. Sequence analysis of the amplicons confirmed the results of the nested-PCR. The presence of anti-Chlamydiaceae antibodies in more than half of the aborting animals (P < 0.002) and the detection of Chlamydophila agents in several fetal organs and in the vaginal swabs are consistent with the history of abortions observed in the herd and suggest an abortifacient role by Chlamydophila spp. in water buffalo (B. Bubalis) herds.     

In Vitro Susceptibility of Chlamydia pecorum to Macrolides,Tetracyclines, Quinolones and β-Lactam

The in vitro susceptibility of Chlamydia pecorum to two macrolides (clarithromycin and erythromycin), two tetracyclines (doxycycline and minocycline), two quinolones (ofloxacin and ciprofloxacin) and one β-lactam (ampicillin) was determined. The MICs were 0.004 to 0.008 μg/ml for clarithromycin, 0.008 to 0.031 μg/ml for doxycycline and minocycline, 0.063 to 0.125 μg/ml for erythromycin, 0.25 to 0.5 μg/ml for ofloxacin and 0.25 to 1.0 μg/ml for ciprofloxacin. The MIC for ampicillin was greater than 1,024 μg/ml. The results show clarithromycin and doxycycline are the two most effective drugs against C. pecorum.     

Diagnosis of ovine enzootic abortion using an indirect ELISA (rOMP91B iELISA) based on a recombinant protein fragment of the polymorphic outer membrane protein POMP91B of Chlamydophila abortus

Chlamydophila abortus is of major economic importance worldwide as one of the principal causes of abortion in sheep. Serological diagnosis of infection by the complement fixation test (CFT) is complicated by false positive reactions resulting from cross-reactive antibodies to Chlamydophila pecorum. To improve diagnosis an indirect enzyme-linked immunosorbent assay (iELISA) based on a recombinant protein fragment of the C. abortus polymorphic outer membrane protein POMP91B (rOMP91B iELISA) was assessed using a panel of 281 sera from experimentally and naturally infected sheep. The iELISA performed well, being more sensitive (84.2%) and specific (98.5%) than the CFT. Furthermore, the iELISA was better at differentiating C. abortus- from C. pecorum-infected animals. The new rOMP91B iELISA test will prove a valuable tool for the routine serodiagnosis of C. abortus infection.     

Sensilla on the antennal funiculus of the horse stomach bot fly, Gasterophilus nigricornis

Gasterophilus nigricornis (Loew) (Diptera: Oestridae) is one of the most damaging obligate parasites of equids in Kalamaili, Xinjiang, China. The main olfactory organs of this stomach bot fly are paired antennae that bear microscopic sensillar structures. The external morphology of the antennal funiculus and sensilla of male G. nigricornis were studied using stereopic microscopy and scanning electron microscopy. A cross-sectional view of the funiculus shows it to be triangular, with an anterodorsal surface, a dorsolateral margin and a posteroventral surface. Almost the entire surface of the funiculus is densely covered with microtrichiae. Small patches lacking these microtrichiae appear as depressions or pits in the surface of the funiculus. Six distinct types of sensilla are recorded, including one trichoid, three basiconic, one auriculate and one clavate sensilla. Trichoid sensilla are the most abundant, followed by the basiconic, auriculate and clavate types in descending order. Only auriculate sensilla are found in pits on the funiculus. Distributions of different sensilla types located on the antennal funiculus are provided. These results are compared with equivalent findings in several other fly species. In addition, the possible functions of the various sensilla types are discussed.     

放归普氏野马核心区黑腹胃蝇疫源分布特点

黑腹胃蝇是新疆荒漠草原优势种马胃蝇,对放归至当地普氏野马构成了严重威胁。该蝇将卵产在牧草上并以此感染采食者——马科动物,其幼虫于宿主消化道营寄生生活,至三龄老熟幼虫随宿主粪便进入环境化蛹,成为当年新的疫源。于黑腹胃蝇幼虫两个发生峰期(4—5月和8月)对普氏野马核心区新鲜粪便和黑腹胃蝇幼虫点位进行了调查,采用最小凸多边形法(MCP)和固定核域法(FKE)计算粪便分布范围,并通过核密度分析(KDE)辨识幼虫聚集区域和特征。结果表明：(1)在黑腹胃蝇种群第一、第二峰期,野马采食草场拥有最多的胃蝇幼虫采集占比(57.25%、41.94%),其次是驴道、山坡、水源地区域;(2)最小凸多边形法(95%利用率)和固定核域法(95%利用率、0.006带宽值)均能较好估算野马粪便分布范围,第一峰期普氏野马粪便范围(60—70 km²)稍大于第二峰期(50—60 km²)。在所调查的八个水源地,两个发生高峰期均包含红柳、5号、6号3个水源地,并形成了以水源地为重要支撑点、依托毗邻草场交叉传播的连续、非均匀疫源地分布特点;(3)水源地附近草场具有更高的黑腹胃蝇幼虫...     

Morphology of a newly discovered sensory array on the mouthhooks of Gasterophilus larvae

The distal mouthhooks of Gasterophilus intestinalis (DeGeer) (Diptera: Gasterophilidae) and G. nasalis (Linnaeus) (Diptera: Gasterophilidae) larvae were studied with scanning electron microscopy to determine the morphology of a previously unknown sensory array. The design of the 3rd stage G. intestinalis sensory array was used as the model for comparison to the 2nd stage of this species and the 2nd and 3rd stadia of G. nasalis. Some components forming the sensory array of the 3rd stage G. intestinalis mouthhook were found in 3rd stage G. nasalis. A major difference between these species was the replacement of shallow pits with sensilla-laden troughs in G. nasalis. Second-stage G. intestinalis has a distal mouthhook that varies considerably from the model type, lacking shallow pits and associated peg-like sensilla. By contrast, the sensory array of the 2nd stage G. nasalis larva is the most elaborate yet encountered, with extensive sensilla and troughs. The existence of other types of sensilla on the mouthhook is proposed, as well as a sensory role for the unsculptured surface. Possible uses of the sensory array by the parasite are discussed.     

Modulation of oxygen binding to insect hemoglobins: the structure of hemoglobin from the botfly Gasterophilus intestinalis

Hemoglobins (Hbs) reversibly bind gaseous diatomic ligands (e.g., O2) as the sixth heme axial ligand of the penta-coordinate deoxygenated form. Selected members of the Hb superfamily, however, display a functionally relevant hexa-coordinate heme Fe atom in their deoxygenated state. Endogenous heme hexa-coordination is generally provided in these Hbs by the E7 residue (often His), which thus modulates accessibility to the heme distal pocket and reactivity of the heme toward exogenous ligands. Such a pivotal role of the E7 residue is prominently shown by analysis of the functional and structural properties of insect Hbs. Here, we report the 2.6 A crystal structure of oxygenated Gasterophilus intestinalis Hb1, a Hb known to display a penta-coordinate heme in the deoxygenated form. The structure is analyzed in comparison with those of Drosophila melanogaster Hb, exhibiting a hexa-coordinate heme in its deoxygenated derivative, and of Chironomus thummi thummi HbIII, which displays a penta-coordinate heme in the deoxygenated form. Despite evident structural differences in the heme distal pockets, the distinct molecular mechanisms regulating O2 binding to the three insect Hbs result in similar O(2 affinities (P50 values ranging between 0.12 torr and 0.46 torr).