Two viruses isolated from rodents (Clethrionomys gapperi and Microtus pennsvlvanicus) trapped in St. Lawrence County, New York

Four strains of C. gapperi virus were isolated from 3 Clethrionomys gapperi and 47 strains of Microtus virus from 15 Microtus pennsylvanicus and 1 Mus musculus. One of the Microtus strains was isolated from a pool of 20 mites while the others were from rodent tissues. These agehts were insensitive to ether and sodium desoxycholate, withstood freezing at -70 C for 3 years and lyophilization without loss of titer, and were not killed when heated at 60 C for 1 hour. Their size as determined by filtration was less than 50 mg and greater than 20-35 mmicro. The strains within each group appear to be similar. The illness induced in suckling mice by the C. gapperi agents had a 5-day incubation period followed by prostration and death with a histologic picture of extensive encephalomalacia. The incubation period in mice for the Microtus agents was 9 to 11 days followed by convulsions and death. Histopathology showed meningeal infiltration and necrosis of the molecular layer. No antigenic similarity was detected between the C. gapperi and Microtus viruses by cross complement-fixation test.     

Direct and indirect effects of masting on rodent populations and tree seed survival

Many plant species are thought to benefit from mast seeding as a result of increased seed survival through predator satiation. However, in communities with many different masting species, lack of synchrony in seed production among species may decrease seed survival by maintaining seed predator populations through the intermast cycle. Similarly, masting by different plant species may have different effects on the seed predator community. We conducted a three-year study in a northeastern USA temperate deciduous forest to determine if production of large seed crops by several tree species was synchronous, and if they had similar effects on all small mammal species. We found that red oak mast crops resulted in increased densities of Peromyscus leucopus and P. maniculatus , but had no effect on Clethrionomys gapperi abundance. Conversely, C. gapperi populations, but not Peromyscus populations, appeared to increase in response to a large red maple seed crop. Differences in small mammal abundance resulted in changes in species-specific seed survival: in the year of abundant C. gapperi , experimentally placed red oak acorns had significantly higher survival than in the year of high Peromyscus abundance. Red oak acorn removal was positively correlated with Peromyscus abundance, while red maple seed removal was significantly higher with increased C. gapperi abundance. Thus, species-specific seed production had differential effects on subsequent small mammal abundance, which in turn affected seed survival. We suggest that at the level of the community, even short-term lack of synchrony in production of large seed crops can cause variation in postdispersal seed survival, through differential effects on the community of small mammal seed predators.     

阿拉斯加东南部温带雨林加氏(鼠平)和肯氏鹿鼠密度与微生境利用的相关性

为了验证对阿拉斯加东南部温带雨林中有关栖息地关系的预测,我们于1999年和2000年8-9月以及1999年和2000年4-5月研究了亚历山大群岛加氏(Clethrionomys gapperi)和肯氏鹿鼠(Peromyscus keeni)种群。我们测量了26个植被和结构特征以检验林隙老龄生长林、多时代老龄生长林、采伐前壮龄(23岁)生长林和泥炭混交针叶林的丰富度与微栖息地利用的相关性。微栖息地利用随季节和栖息地而变化,但加氏与林下落叶灌木覆盖度的正相关最显著。肯氏鹿鼠利用的微环境的林地有较少的苔藓,但是林隙与抓获加氏的概率有直接关系。两种鼠在两个季节的密度与林下腐朽的倒木直接相关。春季的肯氏鹿鼠密度说明加氏密度变化的62%,说明肯氏鹿鼠密度变化的89%。我们的结果印证了早期对阿拉斯加东南部肯氏鹿鼠在各种栖息地尤其早期的演替林中兴旺时的研究;但偏离了西部地貌中肯氏鹿鼠种群在晚期演替针叶林中达到最高密度的普遍结论。与北美西北部其它地区的种群不同,加氏能持久生活在上层被砍伐的雨林板块中。泥炭针叶混交林对两种鼠的繁殖种群几乎没有贡献,因而不可能减轻对多产的老龄生长雨林大规模皆伐所造成的影响。     

Baboon Virus Isolate M-7 with Properties Similar to Feline Virus RD-114

A virus (M-7) isolated from baboon placental tissue demonstrates many similarities to endogenous feline virus RD-114. Immunodiffusion analysis shows a group-specific antigen (gs-1) line of identity between M-7 and RD-114. Anti-RD-114 DNA polymerase IgG inhibits M-7 polymerase by 57% compared to 97% for RD-114. M-7 virus has helper activity as demonstrated by rescue of murine sarcoma virus (MSV) from sarcoma-positive leukemia-negative human amnion cells. The host range of the rescued M-7 pseudotype of MSV, MSV (M-7), is similar to that of RD-114 virus. MSV (M-7) is also able to transform baboon cells and causes no detectable transformation of feline cells without addition of helper feline leukemia virus. Interference properties of M-7 and RD-114 virus are identical. Virus-specific neutralizing antisera, although partially cross-reacting, can distinguish MSV (M-7) from MSV (RD-114). These similarities and differences between RD-114 and M-7 viruses are best explained as type-specific differences between two viruses within the same strain.     

Some characteristics of monoclonal antibodies to a British M A V-like isolate of barley yellow dwarf virus

Rat monoclonal antibodies (MAbs) specific for a British F (MAV-like) isolate of barley yellow dwarf virus (BYDV) were produced and studied. In indirect ELISA using an antiserum to BYDV-F to trap virus from infected sap, the MAbs were shown to be specific for MAV-like isolates of BYDV from Britain, USA and Sweden but, in this test, they did not detect PAV-, RPV-, SGV- or RMV- like isolates of BYDV. In similar tests using homologous antisera to trap the viruses, the MAbs did not detect BYDV-PAV or -RPV or two other luteoviruses (potato leafroll and beet western yellows). One of the MAbs (MAFF 2) was partially purified from ascitic fluid, and used successfully in ELISA as a coating antibody and when conjugated to the enzyme alkaline phosphatase. Also, MAFF 2 successfully trapped BYDV-F particles when used to coat electron microscope grids. In indirect ELISA using three MAbs (MAFF 2, MAC 91 and MAC 92) it was possible to type the three major strain groups of BYDV, viz. MAV, PAV and RPV-like strains from Britain, USA and Europe.     

Spatial and temporal distribution of bluetongue and its Culicoides vectors in Bulgaria

Surveillance of Culicoides (Diptera: Ceratopogonidae) biting midges was carried out between 2001 and 2003, at 119 sites within a 50 x 50-km grid distributed across Bulgaria, using light trap collections around the time of peak adult midge abundance. Sentinel and ad hoc serum surveillance of hosts susceptible to bluetongue infection was carried out at around 300 sites between 1999 and 2003. Following the initial incursion of bluetongue virus 9 (BTV-9) into Bourgas province in 1999, affecting 85 villages along the southern border, a further 76 villages were affected along the western border in 2001, with outbreaks extending as far north as 43.6 degrees N. The BTV-9 strain in circulation was found to have a low pathogenicity for Bulgarian sheep populations, with less than 2% of susceptible individuals becoming sick and seroconversions detected up to 30 km from recorded outbreaks in the south. The major Old World vector Culicoides imicola Kieffer was not detected among over 70,000 Culicoides identified in summer collections, suggesting that BTV-9 transmission in Bulgaria was primarily carried out by indigenous European vectors. The most likely candidates, the Palaearctic species complexes - the Culicoides obsoletus Meigen and C. pulicaris L. complexes - were widespread and abundant across the whole country. The C. obsoletus complex represented 75% of all individuals trapped in summer and occurred in high catch sizes (up to 15,000 individuals per night) but was not found across all outbreak sites, indicating that both Palearctic complexes probably played a role in transmission. Within the C. pulicaris complex, only C. pulicaris s.s., C. punctatus Meigen and C. newsteadi Austen were sufficiently abundant and prevalent to have been widely involved in transmission, whilst within the C. obsoletus complex most trapped males were C. obsoletus s.s. Adult vectors were found to be largely absent from sites in west Bulgaria for a period of at least 3 months over winter, which, taken along with the spatiotemporal pattern of outbreaks in the region between years, indicates the virus may be overwintering here by an alternative mechanism - either by covert persistence in the vertebrate host or possibly by persistence in larval stages of the vector.     

Equilibrium centrifugation studies of hepatitis C virus: evidence for circulating immune complexes.

The buoyant density of hepatitis C virus (HCV), with high in vivo infectivity (strain H) or low in vivo infectivity (strain F), was determined by sucrose gradient equilibrium centrifugation. Viral RNA of strain H was detected in fractions with densities of < or = 1.09 g/ml (principally approximately 1.06 g/ml), while that of strain F was found in fractions with densities of approximately 1.06 and approximately 1.17 g/ml. The observed difference was confirmed by differential flotation centrifugation; in NaCl solution with a density of 1.063 g/ml, most of the HCV RNA of strain H was detected in the top fraction, while that of strain F appeared in the bottom. The same relationship between buoyant density and infectivity was observed in flotation centrifugation experiments with other HCV strains. In immunoprecipitation experiments with anti-human immunoglobulin, HCV (as measured by HCV RNA) was precipitated from the samples with low infectivity and high density but not from those with high infectivity and low density. Examination of serial sera from a chimpanzee infected with HCV revealed parallel changes in the buoyant density and immunoprecipitability of HCV-associated RNA during the course of infection. These data suggest that HCV is bound to anti-HCV antibodies as antigen-antibody complexes in chronic hepatitis C.     

Inhibitory effect of oregano and thyme essential oils on moulds and foodborne bacteria

The essential oil of oregano ('origanum oil'; thymol type oil from Origanum vulgare) inhibited completely the mycelial growth of Aspergillus niger and A. flaous at 400 μg/ml, while A. ochraceus was inhibited at 600 μg/ml. At 700 μg/ml, thyme oil inhibited the mycelial growth of A. flavus and A. niger but not that of A. ochraceus . Fungal spore germination was inhibited by 600 μg/ml of origanum oil and (with the exception of A. ochraceus) by 700 μg/ml of thyme oil. Under aerobic conditions, the essential oils of oregano (250 μg/ml) and thyme (350 μg/ml) inhibited to some extent the growth of Staphylococcus aureus and Salmonella typhimurium. Pseudomonas aeruginosa was not affected by either oregano or thyme oil at concentrations up to 500 μg/ml. The origanum oil was very effective against Campylohacter jejuni and Clostridiurn sporogenes and thyme oil was very effective against C. jejuni. The antagonistic effect of the two oils on Staph. aureus and Salm. typhimuriutn was greatly enhanced when those organisms were incubated in atmospheres of low oxygen tensions     

Environmental factors and nutritional utilization patterns affect niche overlap indices between Aspergillus ochraceus and other spoilage fungi

The total number, and the type of C source utilized in common by an ochratoxin producing strain of Aspergillus ochraceus and six other spoilage fungi varied with the range used (95, Biolog GN plates, or 18 major C sources found in maize). The niche size and niche overlap index (NOI) were markedly influenced by water availability (water activity, aw) and temperature. With freely available water (0.995 aw) there was ecological similarity between A. ochraceus, A. alternata, A. candidus and A. flavus, all exploiting the same sources at both 18 and 30 degrees C. However, under water stress, A. ochraceus had a low NOI compared with A. niger, A. flavus and A. candidus, and Eurotium spp., suggesting that there was very little niche overlap in utilization of different C sources. The NOIs were not directly related to growth rates. Niche overlap between species was shown to be in a state of flux and significantly influenced by environmental factors. This may have important implications in understanding ecological interactions, and dominance by species in a community, and for pre-emptive exclusion of specific species.     

Isolation and characterization of a denitrifying monocrotophos-degrading Paracoccus sp. M-1

A bacterium strain, which is capable of degrading monocrotophos, was isolated from sludge collected from the bottom of a wastewater treatment system of a chemical factory, and named M-1. On the basis of the results of the cellular morphology, physiological and chemotaxonomic characteristics and phylogenetic similarity of 16S rDNA gene sequences, the strain was identified as a Paracoccus sp. The ability of the strain to mineralize monocrotophos was investigated under different culture conditions. Other organophosphorus insecticides and amide herbicides were also degraded by M-1. The key enzyme (s) involved in the initial biodegradation of monocrotophos in M-1 was shown to be a constitutively expressed cytosolic protein. The addition of M-1 (10(6) CFU g(-1)) to fluvo-aquic soil and a high-sand soil containing monocrotophos (50 mg kg(-1)) resulted in a higher degradation rate than that obtained from noninoculated soil. This microbial culture has great potential utility for the bioremediation of wastewater or soil contaminated with organophosphorus pesticides and amide herbicides.     

温度对假单胞rsmA突变株M-18R合成Plt和PCA的区别性影响

次生代谢物阻遏蛋白(Repressor of secondary metabolite,Rsm)A是一种全局性调控因子,与mRNA的RBS结合,转录后水平上抑制基因翻译。运用同源重组技术,构建了假单胞茵(Pseudomonas sp．)M-18的rsmA突变菌株M-18R。在37℃、28℃恒温和短期升温(37℃、4h培养,转28℃继续培养)条件下,比较野生株M-18和突变株M-18R生物合成藤黄绿菌素(Plt)和吩嗪-1-羧酸(PCA)的量。在37℃条件下,M-18和M-18R合成这两种抗生物质的能力几乎受到完全抑制。在28℃条件下,M-18R合成P11的量约为野生型M-18的10倍,达到270μg／mL,但是合成PCA的量仅为野生型的50％。经短期升温培养,M-18的Plt合成量明显下降,PCA产量降低不显;相反,M-18R合成Plt的量达到400μg／mL,但PCA产量的变化仍不明显。推测,M-18菌株细胞内存在着某种与RsmA相关联的温度敏感因子,在RsmA缺失条件下,作为专一性激活剂促进Plt的生物合成,但是,并不参与对PCA合成的调控。     

Numbers of litters, litter size and survival in two species of microtines at two elevations

The number of litters, litter size and survival of Clethrionomys gapperi and Microtus pennsylvanicus were documented at two elevations (1450 m; 2240 m) in the Canadian Rocky Mountains to determine if those life history traits changed with the length of the breeding season. The high elevation was colder, had fewer frost-free days, and more precipitation than the low elevation. The length of the breeding season of C. gapperi and M. pennsylvanicus averaged 8 d and 34 d longer, respectively, at the low elevation than at the high elevation. In both species and at both elevations, most females survived only long enough to produce one or two litters. Litter size was significantly greater at the high elevation than at the low elevation in M. pennsylvanicus , but not different in C. gapperi. Overall, nestling survival was greater at the low elevation than at the high elevation in M. pennsylvanicus , but not different in C. gapperi. Summer and winter survival rates were not different between elevations in either species. Differences in litter size and nestling survival between elevations in M. pennsylvanicus may be related to differences in habitat quality.     

Behavioral types of predator and prey jointly determine prey survival: potential implications for the maintenance of within-species behavioral variation

Recent studies in animal behavior have emphasized the ecological importance of individual variation in behavioral types (e.g., boldness, activity). Such studies have emphasized how variation in one species affects its interaction with other species. But few (if any) studies simultaneously examine variation in multiple interacting species, despite the potential for coevolutionary responses to work to either maintain or eliminate variation in interacting populations. Here, we investigate how individual differences in behavioral types of both predators (ocher sea stars, Pisaster ochraceus) and prey (black turban snails, Chlorostoma funebralis) interact to mediate predation rates. We assessed activity level, degree of predator avoidance behavior, and maximum shell diameter of individual C. funebralis and activity levels of individual P. ochraceus. We then placed 46 individually marked C. funebralis into outdoor mesocosms with a single P. ochraceus and allowed them to interact for 14 days. Overall, predator avoidance behavior and maximum shell diameter were positively associated with survival for C. funebralis. However, the effects of these traits depended on the predator's behavioral type: greater predator avoidance behavior was favored with active P. ochraceus, and low predator avoidance behavior was favored with inactive P. ochraceus. We argue that, even in two-species interactions, trait variation in heterospecifics could be an important factor maintaining trait variation within populations.     

Historical biogeography at the crossroads of the northern continents: molecular phylogenetics of red-backed voles (Rodentia: Arvicolinae)

Evolutionary relationships of red-backed voles and their relatives were examined and used to test biogeographic hypotheses. Sequences of the mitochondrial cytochrome b gene were obtained for 25 individuals representing Alticola macrotis, Clethrionomys californicus, C. gapperi, C. glareolus, C. rutilus, and C. rufocanus. These were combined with 21 partial sequences from GenBank for C. regulus, C. rex, C. rufocanus, C. rutilus, Eothenomys imaizumii, E. melanogaster, Phaulomys andersoni, and P. smithii. Complete sequences of three species of Microtus (M. montanus, M. oeconomus, and M. pennsylvanicus), representative species of other arvicoline genera (Myopus, Synaptomys, Arvicola, Ellobius, Ondatra, Lemmus, Dicrostonyx, and Phenacomys), and a sigmodontine representative (Peromyscus) were included as outgroups. We used maximum parsimony, maximum likelihood, distance, and Bayesian based methods and conducted statistical tests on proposed hypotheses of phylogenetic relationships and biogeographic histories. A close relationship of species representing the genera Alticola, Clethrionomys, and Eothenomys was supported (Clethrionomyini); however, the genus Clethrionomys was paraphyletic with respect to both Alticola and Eothenomys. Three major clades were identified as Asian (Eothenomys andersoni, E. smithii, C. rex, C. regulus, and C. rufocanus), Trans-beringian (Alticola macrotis, C. californicus, C. gapperi, C. glarelolus, and C. rutilus), and Taiwanese (E. melanogaster). These results are consistent with the fossil record which indicates an initial diversification in Asia followed by colonization of the Nearctic on at least two occasions. The holarctic species, C. rutilus, appears to have either reinvaded Asia from North America or colonized North America more recently (late Pleistocene) than the two species of Clethrionomys (C. gapperi and C. californicus) that are endemic to North America (early to mid-Pleistocene). Finally, C. gapperi, appears to be comprised of an eastern and a western species, the former with affinities to the Asian C. glareolus and the latter more closely related to C. californicus.     

Seasonal regulation in fluctuating small mammal populations: feedback structure and climate

We studied fluctuating populations of six small mammal species in the Appalachian Plateau of Pennsylvania, USA for 20 yr. We analyzed the feedback structure of these species using statistical time series models for spring and autumn abundances. All species showed a seasonal density-dependent structure, and in five of them first-order feedbacks were dominant in winter and summer. Instead, southern red-backed voles ( Clethrionomys gapperi ) showed a different feedback structure during winter and summer. In three species ( C. gapperi , Napaeozapus insignis and Peromyscus maniculatus ), environmental factors were more important during summer, while the opposite pattern was found in Blarina brevicauda and Tamias striatus . Snowfall influenced positively the winter population growth rates of southern red-backed voles, white-footed mice, woodland jumping mice and eastern chipmunks. We found seasonal differences in the effects of the small mammals assemblage on population growth rates of the two Peromyscus species. The common feedback structure between seasons observed in most of the species, particularly among voles and mice, points to a different feedback structure from northern cyclic small mammals. We conclude that a seasonal feedback structure dominated by intra- and inter-specific competitive interactions may be at the basis of the population dynamics of these species.     

Improved β-glucan yield using an Aureobasidium pullulans M-2 mutant strain in a 200-L pilot scale fermentor targeting industrial mass production

β-(1→3)-D-glucans with β-(1→6)-glycosidic linked branches are known to be immune activation agents and are incorporated in anti-cancer drugs and health-promoting supplements. β-Glucan concentration was 9.2 g/L in a 200-L pilot scale fermentor using mutant strain Aureobasidium pullulans M-2 from an imperfect fungal strain belonging to A. pullulans M-1. The culture broth of A. pullulans M-2 had a faint yellow color, whereas that of the wild-type had an intense dark green color caused by the accumulation of melanin-like pigments. β-Glucan produced by A. pullulans M-2 was identified as a polysaccharide of D-glucose monomers linked by β-(1→3, 1→6)-glycosidic bonds through GC/MS and NMR analysis. When a conventional medium was used in the culture of A. pullulans M-2 in a 3-L jar fermentor, β-glucan concentration was 1.4-fold that produced by the wild-type. However, when a medium optimized by statistical experimental design was used with dissolved oxygen at 10%, the β-glucan concentration was 9.9 g/L with a yield of 0.52 (g β-glucan/g consumed sucrose), 2.9-fold that of the wild-type. This level of productivity was reproduced when the fermentation was scaled up 200-L. The industrial production of high β-glucan without melanin-like pigments is highly expected, as a health-promoting supplement or functional food.     

Purification of carrot red leaf virus and evidence from four serological tests for its relationship to luteoviruses

Carrot red leaf virus (CRLV) was purified from infected chervil by centrifuging whole plant extracts at low speed and incubating the resuspended pellets with Driselase; the digest was then treated with 1% (v/v) Triton X-100 and the virus concentrated by centrifugation twice at high speed through a layer of 20% sucrose. The preparations (about 1 μg virus/g tissue) contained isometric particles c. 25 nm in diameter which formed a single u.v.-absorbing component in sucrose density gradients. Chervil seedlings exposed to aphids (Cavariella aegopodii) that had been injected with or had fed on fractions from the u.v.-absorbing zone developed typical symptoms of infection with CRLV. CRLV particles had a sedimentation coefficient (s_20,w) of 104 S, buoyant density in CsCl of 1.403 g/cm³ and A₂₆₀/A₂₈₀ of 1.62. Antiserum with a gel-diffusion titre of 1/512 was obtained from a rabbit injected intradermally with 100 μg purified virus. CRLV was detected by immunosorbent electron microscopy and enzyme-linked immunosorbent assay in extracts of the petioles and leaf midribs of infected chervil and in groups of five to 20 viruliferous C. aegopodii. Analysis of antiserum/virus reactions by density gradient centrifugation showed that CRLV is distantly related to all luteoviruses tested; its relationships were closest to barley yellow dwarf virus (RPV strain), and perhaps also to beet western yellows virus, more distant to tobacco necrotic dwarf, potato leafroll and bean leafroll viruses, and very distant to barley yellow dwarf (MAV strain) and soybean dwarf viruses. Some of these relationships were detected by double diffusion in agarose gels and by electron microscopy of antiserum/virus mixtures. Immunosorbent electron microscopy detected all these relationships but suggested that CRLV was more closely related to tobacco necrotic dwarf and potato leafroll viruses than to barley yellow dwarf virus (RPV strain). The results show that CRLV should be considered a definitive member of the luteovirus group, and provide confirmation of recent evidence that potato leafroll virus is a luteovirus.     

Isolation of the Lyme disease spirochete from mammals in Minnesota

Lyme disease spirochetes were isolated from the kidneys of two Peromyscus spp. trapped in Minnesota in September and October 1983. No spirochetes were isolated from white-tailed deer (Odocoileus virginianus), red backed voles (Clethrionomys gapperi), or shrews (Sorexy cinereus and Blarina brevicauda). This is the first report of the isolation of the Lyme disease spirochete from the midwestern United States and isolations from these animals, which were free of ticks, suggest that the Lyme disease spirochete may persist in animal organs for months.     

Specific immunological response against the granulosis virus of the codling moth (Cydia pomonella) in woodmice (Apodemus sylvaticus): field observations

Antibodies specific to the granulosis virus (GV) of the codling moth (Cydia pomonella) were detected by indirect ELISA in sera of woodmice (Apodemus sylvaticus) trapped in an apple orchard sprayed by tractor-drawn mist blower with 2 × 10¹³ virus granules C. pomonella GV/ha. Such antibodies were absent from sera of mice trapped before virus had been applied. Virus antigens were detected by double antibody sandwich ELISA on the ground in and around the orchard and also in the faeces of woodmice inhabiting the orchard, but did not persist from one year to another. Some, but not all, of the samples containing virus antigens were found to be infective.     

Cloning and expression of gene encoding a novel endoglycoceramidase of Rhodococcus sp. strain C9

Endoglycoceramidase (EGCase) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides of various glycosphingolipids. We previously reported that the Asn-Glu-Pro (NEP) sequence is part of the active site of EGCase of Rhodococcus sp. strain M-777. This paper describes the molecular cloning of a new EGCase gene utilizing the NEP sequence from the genomic library of Rhodococcus sp. strain C9, which was clearly distinguishable from M-777 by 16S rDNA analysis. C9 EGCase possessed an open reading frame of 1,446 bp encoding 482 amino acids, and showed 78% and 76% identity to M-777 EGCase II at the nucleotide and amino acid levels, respectively. Interestingly, C9 EGCase showed the different specificity to the M-777 enzyme: it hydrolyzed b-series gangliotetraosylceramides more slowly than the M-777 enzyme, whereas both enzymes hydrolyzed a-series gangliosides and neutral glycosphingolipids to the same extent.