狄斯瓦螨和蜜蜂病毒对意大利蜜蜂蜂群越冬的影响

越冬期是蜂群损失最主要的阶段.通过比较分析45个意大利蜜蜂Apis mellifera ligustica蜂群在繁殖越冬蜂前的狄斯瓦螨Varroa destructor寄生率和病毒感染情况、越冬表现及越冬期存活蜂群的病毒感染情况等,探究与越冬期蜜蜂健康紧密相关的影响因素.结果表明,繁殖越冬蜂前蜂群的狄斯瓦螨寄生率与蜜蜂...     

Serological comparison of polyhedron protein and virions from four nuclear polyhedrosis viruses of plusiine larvae (Lepidoptera: Noctuidae)

Purified polyhedron proteins and purified, ultrasonicated virions of four nuclear polyhedrosis viruses (NPVs), separable into two morphologic groups of singly and multiply embedded virion types (SEVs and MEVs), were investigated by immunodiffusion and immunoelectrophoresis. The four viruses were Pseudoplusia includens SEV, Trichoplusia ni SEV, T. ni MEV, and Autographa californica MEV. In immunodiffusion, SEV polyhedron proteins formed two precipitin bands with antiserum to SEV polyhedron proteins, while MEV polyhedron proteins formed only one. All four proteins formed one precipitin band with antiserum to MEV polyhedron protein, with a spur between SEV and MEV proteins. In immunoelectrophoresis, mobilities of SEV proteins were significantly different from those of MEVs. Precipitin arc patterns were similar to those in immunodiffusion when electrophoresis was carried out at 4 C; at room temperature, a single arc of precipitation formed with all four proteins. SEV virions formed five possibly identical precipitin bands in immunodiffusion with antiserum to SEV virions. MEV virions formed three possibly identical precipitin bands when reacted with antiserum to MEV virions. Little or no cross-reactions were observed between SEV and MEV virions or between virions and polyhedron proteins. In immunoelectrophoresis, SEV virions formed three precipitin arcs in reactions with SEV antisera and none with MEV antisera; MEV virions formed two arcs with MEV antisera and none with SEV antisera. When antisera were subjected to electrophoresis, five arcs were formed by SEVs and three by MEVs in homologous systems, and none were formed in heterologous systems.     

Invasive ants carry novel viruses in their new range and form reservoirs for a honeybee pathogen

When exotic animal species invade new environments they also bring an often unknown microbial diversity, including pathogens. We describe a novel and widely distributed virus in one of the most globally widespread, abundant and damaging invasive ants (Argentine ants, Linepithema humile). The Linepithema humile virus 1 is a dicistrovirus, a viral family including species known to cause widespread arthropod disease. It was detected in samples from Argentina, Australia and New Zealand. Argentine ants in New Zealand were also infected with a strain of Deformed wing virus common to local hymenopteran species, which is a major pathogen widely associated with honeybee mortality. Evidence for active replication of viral RNA was apparent for both viruses. Our results suggest co-introduction and exchange of pathogens within local hymenopteran communities. These viral species may contribute to the collapse of Argentine ant populations and offer new options for the control of a globally widespread invader.     

A study on the possible utilization of immunodiffusion and immunofluorescence techniques as the diagnostic methods for American foulbrood of honeybees (Apis mellifera)

Four types of antisera were obtained from rabbits hyperimmunized with either spores or vegetative rods from two strains of the American foulbrood pathogen, Bacillus larvae. The specificity and sensitivity of these antisera were tested with immunofluorescence and immunodiffusion methods. No cross-reactions were observed between the antisera and other different species of Bacillus or different genera of bacteria. The specificity was not found between the antisera and two strains of B. larvae although stronger fluorescent intensity was observed between the antiserum and its corresponding strain of antigen in the immunofluorescence tests. Eight samples of 1- to 2-day-old larvae, 3- to 4-day-old larvae, decayed tissue, and dry remain, collected from eight infected colonies, were tested against antisera by the immunofluorescence and the immunodiffusion methods. The results indicated that both methods are sensitive and specific for making diagnosis of field samples of American foulbrood of honey bees.     

A serological comparison of some species of microsporida.

Double immunodiffusion techniques were used to investigate the taxonomic relationships between six different microsporidian isolates. Microsporidia used included Nosema bombycis, N. algerae, N. plodiae, and three organisms morphologically similar to N. necatrix. Antigens were extracted from spores after disruption in an MSK Braun cell homogenizer. Cross-reactions were seen between N. plodiae and two of the N. necatrix isolates, while the third N. necatrix, N. bombycis, and N. algerae were antigenically unrelated. One of the N. necatrix isolates revealed temperature-related antigenic differences, but no antigenic differences resulted from aging spores for 5 months before disruption.     

The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses'' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.     

Characterisation and serological relationships of strains of Kashmir bee virus

Isolates of novel strains of Kashmir bee virus (KBV) were obtained from field-collected dead adults of Apis mellifera from honey bee colonies in Canada and Spain. They differed from other strains of KBV in their tendency to aggregate in dilute buffer solution and in containing only three proteins when analysed by SDS-polyacrylamide gel electrophoresis compared with five proteins resolved in the type strain of KBV from Apis cerana in India and six proteins in KBV strains from South Australia and New Zealand. Immunodiffusion tests and Western blotting studies indicated that the five virus isolates were serologically related and all were related to acute paralysis virus (APV). The world distribution of KBV strains and their apparent relationship with APV are discussed.     

RNA viruses as vectors for the expression of heterologous proteins

RNA viruses comprise a wide variety of infectious agents, some of which are the cause of disease in humans, animals, and plants. Recombinant DNA technology is now making it feasible to modify these genomes and engineer them to express heterologous proteins. Several different schemes are being employed that depend on the genome organization of the virus and on the strategy of replication of the particular virus. Several different examples are illustrated and potential uses as well as possible problems are discussed. In the future reverse genetics may convert some of these viruses from agents of disease to agents of cure.     

Occurrence and relative incidence of viruses infecting papaya in Venezuela

A survey of the main papaya (Carica papaya L.) production fields in Venezuela during 1997, indicated that crops were heavily affected with various virus‐like symptoms. A total of 745 samples from papaya plants showing symptoms suggestive of virus infection were collected and analysed using electron microscopy and enzyme‐linked immunosorbent assay (ELISA). Papaya ringspot virus (PRSV) and Papaya mild yellowing virus (PMYV) were the most frequently found viruses, which also occurred, in mixed infections. Rhabdovirus‐like particles were found only in samples collected in Distrito Federal (D.F). Papaya mosaic virus (papMV) and Tomato spotted wild virus (TSW V) were not detected during the survey.     

Multiplex RT-PCR with broad-range primers and an exogenous internal amplification control for the detection of honeybee viruses in bumblebees

Bumblebees are commercially reared and transported worldwide mainly for pollination of greenhouse tomatoes. Three honeybee viruses have been reported in bumblebees: Acute bee paralysis virus, Kashmir bee virus and Deformed wing virus. We developed a multiplex RT-PCR with primers designed on highly conserved regions of the RNA-dependent RNA polymerase in order to detect a maximum range of viral variants. Rearing facilities and governmental organizations can now thoroughly screen bumblebee colonies with a cost-effective technique with an integrated internal amplification control (IAC) implementable in laboratories that strive for International Organization for Standardization (ISO) certification.     

A comparative study of three closely related cytoplasmic polyhedrosis viruses

A cytoplasmic polyhedrosis virus (CPV) from the pine caterpillar, Dendrolimus spectabilis, was compared with Japanese isolates of closely related viruses from the silkworm, Bombyx mori, and gypsy moth, Lymantria dispar. The sizes of the viral RNA genome segments were almost identical, although the CPVs from D. spectabilis and L. dispar could be distinguished from the silkworm virus by a small size difference (0.03 × 10⁶ daltons) in one segment. The same viruses were also distinguishable by RNA homology differences of 25–50% measured by the reannealing of ³H-labeled single-stranded viral messenger RNA (synthesized in vitro) to heat-denatured viral double-stranded RNA. Antigenic differences were also detected by gel immunodiffusion tests. CPVs of D. spectabilis and L. dispar were indistinguishable by these criteria.     

云南食用蜜蜂种类调查及鉴定

在云南,人们有食用蜜蜂幼虫和蛹的习俗。市场上销售的蜜蜂幼虫和蛹多经热水烫过或煮过,因而缺少成虫形态,俗名易混用,食用种类长期未予明确。本文在市场调查的基础上,收集其中部分样品,通过形态学和mtDNA的COI基因部分序列初步鉴定其分类地位,以明确食用种类。结果表明,云南蜜蜂的食用集中在横断山脉南部地区,接受人群广泛,食用方式多样;云南食用本地分布的全部5种野生蜜蜂,包括东方蜜蜂(Apis cerana)、小蜜蜂(A.florea)、大蜜蜂(A.dorsata)、黑大蜜蜂(A.laboriosa)和黑小蜜蜂(A.andreniformis);基于COI序列构建的邻接法(NJ)和最大简约法(MP)系统发育树支持现有的蜜蜂属系统发育关系;现有的食用蜜蜂大多采集自野外,亟待开展遗传资源保护。本研究为食用蜜蜂的鉴定提供了数据支持,也为蜜蜂自然资源的利用和保护提供参考。     

Cancer-associated antigens and antigen arrays in serological diagnostics of malignant tumors

The appearance of antibodies to cancer-associated antigens in biological fluids (particularly, in blood sera) of cancer patients is now a well-established fact, and their detection by immunochemical methods is a promising approach to diagnostics of malignant neoplasms. In this review, we consider some immunobiological aspects of the most extensively studied cancer-associated B-cell antigens, various applications of autoantibodies as cancer biomarkers, and prospects for the use of antigen arrays for improving diagnostic sensitivity.     

Poikilotherm vertebrate cell lines and viruses: A current listing for fishes

Summary A survey of the literature and of work being done in vertebrate cell culture shows that there are currently in existence and available to investigators some 61 cell lines representing 17 families and 36 species of fish. The literature of fish virology shows that at least 17 fish viruses have been isolated and that at least 15 others have been visualized by electron microscopy. A minimum of four major virus groups—rhabdovirus, orthomyxovirus, iridovirus and herpesvirus—are known from fish. Original references, key reviews and sources of cell lines are given.     

FTIR spectroscopic method for detection of cells infected with herpes viruses

Microscopic FTIR spectroscopy was used to investigate the spectral differences between normal cells in culture and cells infected with various members of the herpes family of viruses [Herpes simplex (HSV) and Varicella zoster (VZV)]. The main objective of this study is to evaluate the possibility of developing microscopic FTIR spectroscopy as a sensitive assay for the detection of herpetic infections at their early stages. The advantage of this method over conventional FTIR spectroscopy is that it facilitates inspection of restricted regions of tissue. Our results showed significant and consistent differences between all normal and HSV or VZV infected cells that were tested. Detectable and significant spectral differences between normal and infected cells are seen as early as 24 h postinfection, but the damage of the cells (cytopathic effect), caused by the infecting virus, can be seen by optical microscope observations at only 3 days postinfection. An impressive increase in the levels of vital cellular metabolites was seen in the herpes virus infected cells compared to normal cells. It seems that this spectral behavior is unique for infection with herpes viruses, because when these cells were infected with other viruses from different families like retroviruses, a considerable decrease in the levels of vital cellular metabolites was seen in infected cells compared to normal cells. Cluster analysis performed on FTIR mass chromatography yielded 100% accuracy in classifying control uninfected and VZV or HSV infected cells. Our data strongly support the possibility of developing FTIR microscopy as a diagnostic method for early detection of herpetic infections.     

对蜜蜂性别决定和蜂王级型确立理论的新诠释

从全新视角概貌性解读蜜蜂Apis诸多生殖机制,依据的就是蜂王或产卵工蜂的卵子发生、性别决定假说、性位点研究以及蜂王级型确立机制。这种不同于单倍-二倍性性别决定机制的新诠释,适用于明确性别决定机制、简化定向育种方法、减少比较性实验的难度和时间、实施亲子鉴定和抽象并量化种群的概念。     

Characterization of monoclonal antibodies identifying type and strain-specific epitopes of human immunodeficiency virus type 1

Summary Several hybridoma cell lines were raised against the highly cytopathic Zairian isolate of Human Immunodeficiency Virus (HIV), HIV1-NDK.The specificity of the secreted monoclonal antibodies (mAb) was demonstrated by immunoblotting, radioimmunoprecipitation and immunofluorescence. Two hybridoma cell lines secreted mAb reacting with independent epitopes of the NDK p17 capsid protein and its precursors. One, RL16.24.5, is specific for the NDK isolate whereas the other, RL16.45.1, along with anti-p25 RL16.30.1 mAb, bind all HIV1 isolates but not HIV2. Together with the previously described mAb RL4.72.1 those reagents define lentivirus subfamily (HIV1, HIV2, SIV) type/subtype (HIV1) and strain (HIVI-NDK) specific epitopes expressed on HIVl-NDK core proteins. The last mAb RL16.76.1 binds the env gene products gp160 and gp120.     

A scientific note on the detection of honeybee viruses using real-time PCR (TaqMan) in Varroa mites collected from a Thai honeybee (Apis mellifera) apiary

Bee parasitic mite syndrome is a disease complex of colonies simultaneously infested with Varroa destructor mites and infected with viruses and accompanied by high mortality. By using real-time PCR (TaqMan), five out of seven bee viruses were detected in mite samples (V. destructor) collected from Thailand. Moreover, the results of this study provide an evidence for the co-existence of several bee viruses in a single mite. This is also the first report of bee viruses in mites from Thailand.     

Incidence and Distribution of Grapevine Leafroll-associated viruses in Tunisian Vineyards

Surveys were carried out in Tunisian table grape vineyards for assessing the occurrence and distribution of grapevine leafroll-associated viruses (GLRaVs). Leafroll symptoms were commonly observed in most of the surveyed vineyards. Samples were randomly collected from 712 individual vines for laboratory testing. Enzyme-linked immunosorbent assay (ELISA) tests showed that 81.5% of the vines were infected by one (35.7%) or more (45.8%) viruses. GLRaV-3 was the most widespread virus (76.3%), followed by GLRaV-5 (38.5 %), GLRaV-6 (13.2%), GLRaV-1 (9.1%), GLRaV-2 (6.3%), and GLRaV-7 (0.9%). GLRaV-3 and GLRaV–5, two mealybug-transmissible Ampeloviruses, were present in mixture in 35.9% of samples. The highest infection rate was found in Cape Bon region (81.7 %), where cv. Italia had an infection rate of 79.5%. Superior seedless, the main cultivar in Sidi Bouzid, had 75% infection. GLRaV-6 and -7 were detected for the first time in Tunisia.