Myxosoma cerebralis: In Vitro Sporulation of the Myxosporidan of Salmonid Whirling Disease

SYNOPSIS. Trophozoites and other pre-spore stages of the myxosporidan Myxosoma cerebralis were taken from infected rainbow trout ( Salmo gairdneri ) and cultured in vitro. Cultures eventually yielded mature spores capable of discharging their polar capsules. This is the first report of culture of a myxosporidan.     

Fine Structure of an Unidentified Protozoon in the Epithelium of Rainbow Trout Exposed to Water with Myxosoma cerebralis*

SYNOPSIS. An intracellular protozoon was discovered in the epithelium of young rainbow trout (Salmo gairdneri) exposed for as short a time as 1 hr to water known to contain infective stages of Myxosoma cerebralis. Light- and electron-microscopic examination of this tissue revealed what appeared to be a proliferative stage (presumptive schizont) of a sporozoon; other possible stages in the life cycle were also observed. The relationship of this unidentified protozoon to M. cerebralis remains unresolved.     

Small Subunit Ribosomal RNA Sequences Unite Alternate Actinosporean and Myxosporean Stages of Myxobolus cerebralis the Causative Agent of Whirling Disease in Salmonid Fish

ABSTRACT. The alternating myxosporean and actinosporean stages of the myxozoan parasitc Myxobolus cerebralis (Hofer 1903) from its salmonid fish and aquatic oligochaete hosts, respectively, were compared for sequence homology of the small subunit (18S) ribosomal RNA genes. A 99.8% similarity between the sequences of these two stages was substantially greater than that of M. cerebralis compared to two other Myxobolus sp. from salmonid fish. Our results are the first molecular evidence confirming the alternating stages initially described by Wolf and Markiw [25] for the life cycle of M. cerebralis but found in two different taxonomic classes (Myxosporea and Actinosporea) are indeed forms of the same organism. Sequencing of rRNA genes of the actinosporean stage followed by development of specific primers for DNA amplification of the myxosporean stage, as in our study, should be applied to solve other myxozoan life cycles. Additionally, these approaches will in the future provide useful diagnostic reagents for the detection and study of this important group of fish pathogens.     

Myxosoma cerebralis (Myxozoa: Myxosporea) Etiologic Agent of Salmonid Whirling Disease Requires Tubificid Worm (Annelida: Oligochaeta) in its Life Cycle

Studies of the life cycle of Myxosoma cerebralis showed that development of infectivity did not occur endogenously but that the spore “aging” process required participation of an aquatic tubificid oligochaete. Data suggestive of such involvement were derived from trials in which spores were “aged” in an array of inert, sterilized, pasteurized, or natural aquatic substrates and from examination of aquatic soils from trout hatcheries in which whirling disease was epizootic. The role of the aquatic oligochaete was confirmed two ways. First, signs of whirling disease developed, and M. cerebralis spores were produced in young rainbow trout (Salmo gairdneri) that had been fed oligochaetes harvested from pond soil taken from two hatcheries where whirling disease was epizootic. Second, when containers of pasteurized soil were populated with four genera of oligochaetes–Aeolosoma, Dero, Stylaria, or Tubifex– from a biological supply house, or with tubificid worms from trout hatcheries free of whirling disease, and then seeded with M. cerebralis spores and “aged” for 4 months, whirling disease occurred only in trout held with Tubifex and with hatchery tubificids.     

Primate Conservation: The Prevention of Disease Transmission

We address the strategies to prevent disease transmission from human to non-human primates in natural settings. Some field research methods, such as gaining close proximity for observation, provisioning for habituation, or reintroducing for repopulation, may place primate subjects at risk for acquiring human-carried diseases. Additional risks arise through inadequate waste disposal or nonhygienic conditions of humans residing at the study site. We describe several disease outbreaks at primate field sites, emphasizing the need for proper protocols to diagnose, to treat, and to prevent recurrence. Finding solutions to the disease transmission problem requires effecting change in the behavior and policies of many individuals, including field researchers, veterinarians, human health care providers, park personnel, government officials, local villagers, and tourists. The prevention of exposure to infectious disease is an important, fundamental aspect of primate conservation; the assurance of good health and longevity in wild primate populations is paramount to the more traditional conservation issues of poaching control and forest protection.     

Relating Phylogenetic Trees to Transmission Trees of Infectious Disease Outbreaks

Transmission events are the fundamental building blocks of the dynamics of any infectious disease. Much about the epidemiology of a disease can be learned when these individual transmission events are known or can be estimated. Such estimations are difficult and generally feasible only when detailed epidemiological data are available. The genealogy estimated from genetic sequences of sampled pathogens is another rich source of information on transmission history. Optimal inference of transmission events calls for the combination of genetic data and epidemiological data into one joint analysis. A key difficulty is that the transmission tree, which describes the transmission events between infected hosts, differs from the phylogenetic tree, which describes the ancestral relationships between pathogens sampled from these hosts. The trees differ both in timing of the internal nodes and in topology. These differences become more pronounced when a higher fraction of infected hosts is sampled. We show how the phylogenetic tree of sampled pathogens is related to the transmission tree of an outbreak of an infectious disease, by the within-host dynamics of pathogens. We provide a statistical framework to infer key epidemiological and mutational parameters by simultaneously estimating the phylogenetic tree and the transmission tree. We test the approach using simulations and illustrate its use on an outbreak of foot-and-mouth disease. The approach unifies existing methods in the emerging field of phylodynamics with transmission tree reconstruction methods that are used in infectious disease epidemiology.     

Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asial by RT-PCR

To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.     

Cyclical Transmission of Trypanosoma brucei rhodesiense and Trypanosoma congolense by Tsetse Flies Infected with Culture-Form Procyclic Trypanosomes*

SYNOPSIS. Culture procyclic forms of Trypanosoma brucei rhodesiense and Trypanosoma congolense were fed to Glossina morsitans morsitans through artificial membranes. A very high percentage of the flies so fed produced established midgut infections, a proportion of which went on to develop into mature metacyclic trypanosomes capable of infecting mammalian hosts. The method offers a safe, clean way of infecting tsetse flies with African trypanosomes which reduces the need for trypanosome-infected animals in the laboratory.     

Effects of Fertility Control on Behavior and Disease Transmission in Brushtail Possums

Abstract: Fertility control is currently under development for the control of brushtail possums (Trichosurus vulpecula), one of New Zealand's most serious vertebrate pests. Despite intensive research into various methods for achieving infertility, including immunocon-traception and disrupting endocrine control of reproduction, researchers know little about the potential effects of these methods on the behavior of wild possums. We assessed the effects of surgically imposed sterility, either to block fertilization (tubal ligation) or to disrupt endocrine control of fertility (gonadectomy), by using radiotelemetry on the movement patterns and site fidelity of wild brushtail possums. In addition, we assessed the effect of gonadectomy on the transmission rate of a commonly occurring, directly transmitted pathogen in possums, Leptospira interrogans serovar balcanica (hereafter L. balcanica), to determine the effect of any behavioral changes on possum contact rates. Both tubal ligation and gonadectomy of females did not appear to have any appreciable effect on behavior, with sterilized females having space-use patterns and fidelity to seasonal breeding ranges similar to those of fertile females. However, gonadectomy of male possums resulted in a significant reduction of 42% and 47% in the 95% and 70% isopleth seasonal breeding ranges, respectively. Furthermore, the transmission rate of L. balcanica in gonadectomized male and female possums was reduced by 88% and 63%, respectively, compared with that in fertile male and female possums. Overall, these results suggest that fertility control, either by blocking fertilization (e.g., immunocontraception) or by disrupting endocrine control of reproduction (e.g., gonadotropin-releasing hormone vaccines), is unlikely to have an impact on social organization and behavior of brushtail possums in ways that may compromise the efficacy of fertility control for reducing population density. However, the reduction in the transmission rate of L. balcanica indicates that fertility control that interferes with endocrine control of reproduction is likely to reduce the contact rate between possums. This could have implications for the control of other wildlife diseases requiring direct contact for transmission.     

Risk of Disease Transmission between Conservation Personnel and the Mountain Gorillas: Results from an Employee Health Program in Rwanda

Humans and gorillas share 97% of their genetic makeup which means the risk of disease transmission between the two is potentially high. Humans with high exposure and whose exposure-related activity can most easily be managed are park conservation personnel. In June 2001, the Morris Animal Foundations Mountain Gorilla Veterinary Project initiated a health program for all employees working in Rwandas Parc National des Volcans in collaboration with in-country government and nongovernmental agencies. The goal is to improve the health of conservation personnel and reduce the risk of zoonotic disease transmission between employees and the parks mountain gorillas. Employees annually receive a clinical examination and laboratory testing, and provide a clinical history, In 2002, analyses were performed on the dataset of 127 employees to identify potential risk factors associated with positive laboratory tests. Considering all fecal tests combined, 70.1% were positive for one or more pathogenic organisms. A high percentage (> 80%) tested positive on viral antibody titer testing for various communicable diseases including measles, chickenpox, and hepatitis. On multivariate analysis, the main risk factor for testing positive for any pathogenic organism was use of a pit latrine at home. Vaccination against childhood communicable diseases and improved human waste disposal could be critical control points for preventing disease transmission to mountain gorillas. Program results have been shared with local health officials to aid in their efforts to improve village health and sanitation standards, and with park employers as a basis for ongoing employee health education.The Mountain Gorilla Veterinary Project 2002 Employee Health Group members are listed in Appendix 1. by area of contribution.     

Transmission routes maintaining a viral pathogen of steelhead trout within a complex multi‐host assemblage

This is the first comprehensive region wide, spatially explicit epidemiologic analysis of surveillance data of the aquatic viral pathogen infectious hematopoietic necrosis virus (IHNV) infecting native salmonid fish. The pathogen has been documented in the freshwater ecosystem of the Pacific Northwest of North America since the 1950s, and the current report describes the disease ecology of IHNV during 2000–2012. Prevalence of IHNV infection in monitored salmonid host cohorts ranged from 8% to 30%, with the highest levels observed in juvenile steelhead trout. The spatial distribution of all IHNV‐infected cohorts was concentrated in two sub‐regions of the study area, where historic burden of the viral disease has been high. During the study period, prevalence levels fluctuated with a temporal peak in 2002. Virologic and genetic surveillance data were analyzed for evidence of three separate but not mutually exclusive transmission routes hypothesized to be maintaining IHNV in the freshwater ecosystem. Transmission between year classes of juvenile fish at individual sites (route 1) was supported at varying levels of certainty in 10%–55% of candidate cases, transmission between neighboring juvenile cohorts (route 2) was supported in 31%–78% of candidate cases, and transmission from adult fish returning to the same site as an infected juvenile cohort was supported in 26%–74% of candidate cases. The results of this study indicate that multiple specific transmission routes are acting to maintain IHNV in juvenile fish, providing concrete evidence that can be used to improve resource management. Furthermore, these results demonstrate that more sophisticated analysis of available spatio‐temporal and genetic data is likely to yield greater insight in future studies.     

Focus: Zoonotic Disease: An Ecologically Framed Comparison of The Potential for Zoonotic Transmission of Non-Human and Human-Infecting Species of Malaria Parasite

The threats, both real and perceived, surrounding the development of new and emerging infectious diseases of humans are of critical concern to public health and well-being. Among these risks is the potential for zoonotic transmission to humans of species of the malaria parasite, Plasmodium, that have been considered historically to infect exclusively non-human hosts. Recently observed shifts in the mode, transmission, and presentation of malaria among several species studied are evidenced by shared vectors, atypical symptoms, and novel host-seeking behavior. Collectively, these changes indicate the presence of environmental and ecological pressures that are likely to influence the dynamics of these parasite life cycles and physiological make-up. These may be further affected and amplified by such factors as increased urban development and accelerated rate of climate change. In particular, the extended host-seeking behavior of what were once considered non-human malaria species indicates the specialist niche of human malaria parasites is not a limiting factor that drives the success of blood-borne parasites. While zoonotic transmission of non-human malaria parasites is generally considered to not be possible for the vast majority of Plasmodium species, failure to consider the feasibility of its occurrence may lead to the emergence of a potentially life-threatening blood-borne disease of humans. Here, we argue that recent trends in behavior among what were hitherto considered to be non-human malaria parasites to infect humans call for a cross-disciplinary, ecologically-focused approach to understanding the complexities of the vertebrate host/mosquito vector/malaria parasite triangular relationship. This highlights a pressing need to conduct a multi-species investigation for which we recommend the construction of a database to determine ecological differences among all known Plasmodium species, vectors, and hosts. Closing this knowledge gap may help to inform alternative means of malaria prevention and control.     

Transmission by Laodelphax striatellus Fallen of Rice black‐streaked dwarf virus from Frozen Infected Rice Leaves to Healthy Plants of Rice and Maize

Rice black‐streaked dwarf virus (RBSDV) is transmitted naturally to important crops such as rice, maize, barley and wheat in a persistent manner by the planthoppers, Laodelphax striatellus, Unkanodes sapporona and Unkanodes albifascia. Insect vector transmission tests are the basis for identifying viral incidence, evaluating the resistance of varieties and selecting resistance sources for rice and maize breeding. A simple, rapid and reliable method is described by which virus‐free small brown planthoppers (L. striatellus) acquired RBSDV from frozen infected rice leaves and transmitted it to healthy rice and maize plants. After feeding on frozen infected rice leaves, the planthoppers were tested by RT‐PCR for the presence of virus after 10, 15, and 22 days, respectively. The percentages of RBSDV‐containing insects were 0, 25 and 71.43% of L. striatellus fed on frozen infected rice leaves compared to 0, 28.25 and 71.43% of L. striatellus fed on fresh infected rice leaves, respectively. In transmission tests, three of eight rice seedlings (37.5%) and four of eight maize seedlings (50%) were inoculated by the planthoppers that had fed previously on frozen leaves and had allowed a 22 days latent period and showed typical disease symptoms. As a positive control, four of eight rice seedlings (50%) and four of six maize seedlings (66.67%) became infected. All rice and maize plants expressing disease symptoms were identified as virus‐positive by RT‐PCR. These results indicated that the planthoppers acquired RBSDV from frozen infected leaves and transmitted the virus to healthy plants.     

应用亲和层析法提纯鸡马立克氏病病毒pp38基因重组产物

利用鸡马立克氏病病毒(MDV)Ⅰ型特异单克隆抗体H_(19)致敏Sepharose 4B-CNBr,从感染重组病毒BP38Ⅱ的昆虫细胞中提纯鸡马立克氏病毒pp38基因重组产物,获得良好效果。提纯的蛋白质在SDS-PAGE中表现出一条分子量约为38kDa的蛋白质条带,在免疫印迹试验中该蛋白质条带也能被单克隆抗体H_(19)识别。利用该提纯的重组pp38免疫小鼠,所制备的小鼠抗血清在免疫荧光染色试验中不仅能与感染重组病毒的昆虫细胞Sf9反应,也能和Ⅰ型MDV型感染的鸡胚成纤维细胞反应。     

Structural and Dynamic Analysis of Leaf-Associated Fungal Community of Walnut Leaves Infected by Leaf Spot Disease Based Illumina High-Throughput Sequencing Technology

Leaf-associated microbiota is vital in plant-environment interactions and is the basis for micro-ecological regulation. However, there are no studies on the direct differences in microbial community composition between disease-susceptible and healthy walnut leaves. This study collected five samples of healthy and infected leaves (all leaves with abnormal spots were considered diseased leaves) from May to October 2018. Differences in fungal diversity (Chao1 index, Shannon index, and Simpson index) and community structure were observed by sequencing and analyzing diseased and healthy leaf microbial communities by Illumina HiSeq sequencing technology. The main fungal phyla of walnut leaf-associated were Ascomycota, Basidiomycota, and Glomeromycota. Diversity indices (Shannon and Chao1 index values) of healthy leaves differed significantly in the late stages of disease onset. The results showed that the fungal species that differed considerably between the healthy and infected groups differed, and the fungal species that differed significantly between the healthy and infected groups changed with the development of the leaf disease. Critical control time points were determined by analyzing the population dynamics of pathogenic fungi. Leaf-associated microorganisms are abundant and diverse, and fungal identification and diversity studies are helpful for developing more appropriate walnut management strategies Open in a separate window     

辛伐他丁治疗冠心病高血脂症临床观察

目的观察辛伐他丁对冠心病高血脂症患者血清脂质的影响。方法冠心病高血脂症病人５０例,均有血脂（ＴＣ、ＴＧ、ＨＤＬ－Ｃ、ＬＤＬ－Ｃ或４项中任何一项）异常。服用辛伐他丁,５～２０ｍｇ,每日１次,晚间顿服,疗程８周。服药前后晨起空腹查血脂以作自身对照。结果对高ＴＣ下降有效率达９１．５％,对高ＬＤＬ－Ｃ下降有效率达８２．５％,对高ＴＧ下降有效率达７６．２％,对低ＨＤＬ－Ｃ增高有效率为７２．２％。其中对ＴＣ、ＬＤＬ－Ｃ治疗８周有极显著意义（Ｐ＜０．０１）,,对ＴＧ下降、ＨＤＬ－Ｃ升高８周后有显著意义（Ｐ＜０．０５）。治疗期间不良反应轻微。结论辛伐他丁可作为冠心病患者减少ＣＨＤ事件发病率及住院率的理想药物。     

阴道镜配合病理活检应用于宫颈疾病的诊断价值

目的评价电子阴道镜及宫颈病理活检对宫颈疾病的诊断价值。方法收集我院妇科门诊2003年至2006年宫颈疾病患者行阴道镜检查病行宫颈活检的病理结果进行对照分析。结果电子阴道镜检查后行宫颈活检1060例,病理结果显示:宫颈癌16例,宫颈上皮内瘤样变342例,宫颈炎性病变702例,阴道镜诊断宫颈上皮内瘤样变及宫颈癌的符合率分别为:93.44%和93.75%。结论阴道镜检查配合病理活检诊断宫颈疾病结果及时可靠,尤其对宫颈上皮内瘤样变的早诊断及降低宫颈癌的发生率有重要价值。     

Dry-season Mosquito Breeding Associated with Irrigation in the Northeast Kimberley Region of Western Australia: Potential Impact on Mosquito-borne Disease Transmission

Recent Ross River virus activity prior to the onset of the wet season in the Kimberley region of Western Australia points to an increased dry season mosquito-borne disease risk at Kununurra, compared to other Kimberley towns. This study describes a preliminary investigation into the role of the Ord River Irrigation Area at Kununurra in mosquito production during the dry season. Specifically, the study sought to determine whether the irrigation area has provided the opportunity for year-round breeding of arbovirus vector mosquitoes. A 10-day adult and larval mosquito sampling survey, in August 2003, revealed that mosquito breeding was occurring during the driest month of the year at Kununurra, supporting the hypothesis that mosquitoes can breed year-round. Importantly, significantly larger numbers and percentage of adult Culex annulirostris, an important disease vector in Australia, were collected within the irrigation area (44.6% of total catch) compared to nearby reference (nonirrigated) breeding sites (9.8%) (mean difference 76.2 per trap per night; 95% CI 38.6, 113.7; P <0.001). Larval Cx. annulirostris were also collected at several sites within the irrigation area, whereas none were collected at reference sites. These results indicate that mosquito breeding associated with anthropogenic environmental changes may be responsible for an increased health risk at the end of the dry season. Mosquito management needs to be given a high priority to ensure this potential health risk is not further exacerbated. Several control strategies to reduce breeding of disease vector mosquitoes are identified, primarily focusing on modification of breeding habitats and alteration of irrigation protocols.     

苍白球电毁损对帕金森病大鼠HVSs 振荡波的影响研究

目的:应用直流电核团毁损术毁损帕金森病(PD)大鼠模型的内侧苍白球(GPi),记录其手术前后脑电生理活动的变化,以探讨内苍白球射频毁损术治疗PD的可能机制。方法:成年SD大鼠随机分为GP毁损组、假手术组及正常组。对PD毁损组和假手术组大鼠采用6-羟基多巴胺(6-OHDA)右侧黑质致密部(SNc)、中脑腹侧被盖区(VTA)两点注射法建立PD大鼠模型,并经腹腔注射阿扑吗啡(APO)诱发旋转以对模型建立进行评价。通过多导联宏电极在体脑电生理记录技术对各组大鼠进行右侧(注射侧)大脑皮层M1、M2区脑电及纹状体场电位的连续24小时记录,同时进行视频录像。对GP毁损组大鼠行直流电GPi毁损术,术后4天对各组大鼠均行阿扑吗啡诱导旋转行为检验,继续记录脑电活动,记录数据经频率谱分析及相干分析以揭示各记录位点信号的频率成分以及不同位点神经元集群间的功能连接和同步性。结果:对GP毁损组大鼠毁损术前后在清醒静息状态下的皮层脑电和纹状体场电位有明显改变,术后HVSs(High Voltage Spindles)在持续时间上明显缩短发作次数明显减少;对各组大鼠术后在静息状态下的脑电信号进行对比,GP毁损组大鼠较假手术组的HVSs持续时间和发作频率均减少并接近正常组大鼠水平,相干性分析显示GP毁损组大鼠术后在HVSs频段(5-13Hz)上的相干程度显著小于假手术组。结论:在清醒静息状态下6-OHDA建立的PD大鼠皮层-基底节环路上HVSs持续时间延长发生频率增高,经GP毁损术后其时间缩短发作次数减少同步性降低并接近正常水平,从而改善PD症状,该现象可能解释临床采用苍白球射频毁损术治疗PD的治疗机制。     

Effects of Temperature on Disease Severity in Plants of Subterranean Clover Infected Singly or in Mixed Infection with Bean yellow mosaic virus and Kabatiella caulivora

Many epidemics involve plants infected with more than one pathogen, but few experiments address climate change scenarios that influence mixed infections. This study addresses the interactive effects of co‐infection and temperature on disease development in plants of the annual pasture species subterranean clover (Trifolium subterraneum), which is widely sown in different world regions. Bean yellow mosaic virus (BYMV) and the fungus Kabatiella caulivora are two important pathogens causing considerable production losses in pastures containing this species. Both occur together in such pastures causing a severe necrotic disease when mixed infection occurs. Effects of temperature on symptom expression were investigated in subterranean clover plants infected singly or in mixed infection with these pathogens. Plants were maintained in controlled environment rooms at 18°C, 20°C or 22.5°C after sap inoculation with BYMV. K. caulivora conidia suspensions were inoculated to plants once systemic BYMV symptoms developed. Plants were assessed for three disease assessment parameters, dead petioles numbers, marginal leaflet necrosis and overall plant damage. In general, mixed infection caused most severe symptoms, K. caulivora least severe symptoms, and BYMV symptoms of intermediate severity. In single infections, effects of temperature on disease severity differed between pathogens: BYMV symptoms were most pronounced at 18°C, but K. caulivora induced more severe symptoms at 20°C and 22.5°C. In mixed infections, disease severity generally followed the pattern developed with BYMV alone as temperature increased. Also, synergistic increase in disease severity sometimes occurred at 18°C, but increases were only additive at 20°C and 22.5°C. These results reflected the greater BYMV multiplication detected in infected leaves at 18°C compared with 20°C or 22.5°C. Our findings indicate that in rainfed subterranean clover pastures, as global warming progresses disease severity from infection with BYMV and K. caulivora alone may decline or increase, respectively, and mixed infection with them may become less damaging.