The effects of serial passage of a nucleopolyhedrosis virus through an alternate host system

The multiple-embedded velvetbean caterpillar nucleopolyhedrosis virus (VBC-NPV) ofAnticarsia gemmatalis Hübner was shown to be infectious of a variety of noctuid hosts. The mortality data demonstrated the importance of defining the dosage used in host range analysis. Serial passage of the VBC-NPV through the soybean looper,Pseudoplusia includens, was done to select for host range variants of this NPV. After several passages of the VBC-NPV through this insect the virulence of the progeny virus remained unchanged indicating heterogeneity in the host and not the virus population. However, between the 3rd and 5th serial passage throughPseudoplusia a latent NPV identical to a single-embedded NPV previously associated fromA. gemmatalis was activated. The biological and biochemical characteristics of this isolate demonstrated it to be distinct from the original VBC-NPV. A single passage of this activated virus throughA. gemmatalis resulted in the production of viral progeny having characteristics of the original VBC-NPV. Serial passage of this virus preparation throughP. includens resulted in virus progeny having biological properties associated with both the original VBC-NPV and the activated NPV isolate.     

In situ growth and photosynthetic activity of Cyanobacteria and phytoplankton dynamics after passage through the gut of silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), and Nile tilapia (Oreochromis niloticus)

The growth and photosynthetic activities of Cyanobacteria passed through the gut of silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), and tilapia (Oreochromis niloticus) were compared with those of phytoplankton taken directly from Lake Taihu during a 13-day in situ dialysis culture. After the first 3–5 days of reduced activity after excretion by silver carp and bighead carp, the photosynthetic activity of Cyanobacteria recovered and rose significantly higher (P < 0.01) than levels in the control population, whereas there was a notable reduction of photosynthetic activity after passage through tilapia gut. The phytoplankton biomass showed a 2- to 3-fold increase of growth, and extracellular polysaccharide production was also stimulated after passage through silver carp and bighead carp gut. Chlorophyta fluorescence was detected at much higher levels than that of Cyanobacteria and Bacillariophyta after passage through tilapia gut. Scenedesmus obliqnus and Chlamydomonas sp. contributed much to the growth of the Chlorophyta during the in situ cultivation. However, the total phytoplankton biomass showed a distinct reduction in the tilapia treatment during the culture. The study indicated that Nile tilapia feeding and defecation may help remove Cyanobacteria from the water column and favor a community shift to Chlorophyta.     

Hypervirulent-Host-Associated Citrobacter rodentium Cells Have Poor Acid Tolerance

Enhanced virulence or infectivity after passage through a mammalian host has been reported for a number of enteric food-borne pathogens. Citrobacter rodentium is a mouse pathogen that mimics many aspects of enterohemorrhagic Escherichia coli infection of humans and serves as a useful model for studying virulence mechanisms. Emergence of a hyperinfectious state after passage through mouse gastrointestinal tract was reported for C. rodentium. We wanted to investigate if increased acid tolerance could explain hypervirulence status of C. rodentium. Although we were able to observe hyperinfectious state of C. rodentium upon host passage, the cells were extremely acid sensitive. Growth under mildly acidic conditions (LB-MES, pH 5.5) induced acid tolerance of C. rodentium, but did not improve the organism’s ability to establish infection. Growth under anaerobic environment on fecal components also did not induce hyperinfectious state. Thus, contrary to conventional anticipation, hypervirulent C. rodentium cells were found to be acid sensitive thereby revealing limitations of the role of mouse gastric acidity by itself in elucidating the hypervirulent phenotype.     

A Few-Polyhedra Mutant and Wild-Type Nucleopolyhedrovirus Remain as a Stable Polymorphism during Serial Coinfection in Trichoplusia ni

Few-polyhedra (FP) mutants of nucleopolyhedroviruses (NPVs) are a well-known phenomenon during serial passage of virus in cell culture. Under these circumstances such mutants produce low yields of occlusion bodies (OBs) and poorly occlude virions, but they are selected for through advantageous rates of budded virus replication. Spontaneous insertion of transposable elements originating from host cell DNA into the viral fp25 gene has been shown to be a common cause of the phenotype. A model of NPV population genetics predicts that mutants with these characteristics might persist within stable polymorphisms in viral populations during serial passage of virus in vivo. However, this hypothesis was previously untested, and FP mutants have not been recovered from field isolates of NPVs. We isolated and characterized an FP mutant that arose during routine passage of Autographa californica multinucleocapsid NPV (AcMNPV) in cell culture and identified a transposable element within the fp25 gene. We tracked the fates of coinfecting wild-type and FP mutant AcMNPV strains through serial passage in fifth-instar Trichoplusia ni larvae. The levels of both strains remained stable during successive rounds of infection. We applied the data obtained to a model of NPV population genetics in order to derive the frequency distribution of the multiplicity of cell infection in infected insects and estimated that 4.3 baculovirus genomes per OB-producing cell would account for this equilibrium.     

Multi-year monitoring to facilitate adult salmon passage through a temperate tidal marsh

Adult Chinook salmon, Oncorhynchus tshawytscha, were ultrasonically tagged to monitor their movements past a tidally operated salinity control-gate structure that spans the width of Montezuma Slough, an important migration pathway for anadromous fish in the Sacramento Delta. The structure consists of a section of removable flashboards, three radial gates and a boat lock. Previous studies showed that movement through Montezuma Slough was delayed or hindered for many of the tagged salmon due to installation of the flashboards and the gate closures. Three gate configurations were tested as future mitigation options to facilitate salmon movement through the gates: full open with the boat lock closed; full operation with the boat lock open; and full operation with the boat lock closed. During October to November of 2001–2004, approximately 200 adult fall-run Chinook salmon were tagged annually and monitored for passage success during different gate configurations. Relative to the closed boat lock configuration, the full open configuration had a significantly higher passage rate in 2004 only, and a significantly shorter passage time in 2001. Relative to the closed boat lock configuration, the full operation with the boat lock open configuration was correlated with improved passage rates in 2 out of 4 years, with shorter passage times in 2001, and with longer passage times in 2002. The findings from this study suggest that the open boat lock can provide additional salmon passage at the salinity control gates, but that further multi-year studies using biotelemetry to track fish movement through Montezuma Slough should be addressed in future research.     

Babesia bovis: Proteins of virulent and avirulent parasites passaged through ticks and splenectomized or intact calves

Passage of the avirulent vaccine (K) strain of Babesia bovis (K_A) through either Boophilus microplus ticks, intact calves, or intact calves and then ticks, resulted in two distinct protein and protein antigen profiles as analyzed by two-dimensional gel electrophoresis of biosynthetically labeled proteins and immunoprecipitates. Different degrees of expression of two major acidic antigens of K_A designated Ka₁ (M_r 47,500) and Ka₂ (M_r 43,000) were observed. Ka₁ was apparently lost following passage of K_AB. bovis through intact calves but was strongly represented in the parasite population following a single tick passage. In contrast, passage through ticks of the virulent K_VB. bovis (from which K_A was derived by passage in splenectomized calves) did not lead to strong representation of the Ka₁ protein although there was increased representation of another major acidic protein antigen, designated Kv (M_r 35,000). These data suggest that the previously recognized reversion to a strain-dependent basal antigenic type in the tick vector depends also on intrastrain characteristics such as virulence and strain heterogeneity. The data suggest that K_A is a more heterogeneous population than K_V although cloned isolates are required to establish this point. Comparable syringe passage of another strain of B. bovis, designated C strain, through splenectomized calves resulted in less marked differences between the putative C_A and C_VB. bovis. This may explain the less stable avirulence of C_A compared to K_AB. bovis. Various selection pressures must act, in either the tick or the vertebrate host, on subpopulations in heterogeneous isolates to produce the changes described in protein antigen profiles of B. bovis. The possible relevance of changes in representation of proteins to biological characteristics of B. bovis (such as virulence and tick transmissibility) is discussed.     

The role of individual behavioral traits on fishway passage attempt behavior

Variations in behavioral traits are widely recognized to drive animal behaviors exhibited within a population. However, information on how behavior traits influence behavior in anthropogenically modified habitats is lacking. Many habitats have become highly fragmented as a result of human processes. To mitigate this and improve habitat connectivity, wildlife passes are increasingly employed, with the aim of enabling animals to move freely between habitats. However, wildlife passes (e.g., fishways) are not always effective in achieving passage and it remains uncertain what factors play a role in an individual''s likelihood of passing successfully. This study measured three behavioral traits (boldness, exploration, and activity) in juvenile brown trout (Salmo trutta; n = 78) under field conditions within a river and tested whether these behavior traits influenced both the passage success and the behaviors exhibited during upstream fishway passage attempts. Although behavioral traits were found and collapsed into two behavioral trait dimensions, behavioral traits had low repeatability and so did not contribute to a personality spectrum. Boldness was found to negatively influence the number of passage attempts carried out by an individual and to positively influence passage success, with bolder individuals carrying out fewer attempts and having an increased probability of passage success. No behavioral traits were found to be related to other passage metrics (passage success, Time until First Attempt, and Passage Duration) during the first passage. But all three behavioral traits were significantly negatively related to the changes in passage behaviors at consecutive, successful passage attempts, with bolder, more exploratory and more active individuals passing through a fishway quicker on the second passage than on the first. This study suggests that bolder and more active individuals may perform better during fishway passage attempts, particularly within rivers where multiple barriers to movement exist.     

Selection of Bacteria with Favorable Transport Properties Through Porous Rock for the Application of Microbial-Enhanced Oil Recovery

This paper presents a bench-scale study on the transport in highly permeable porous rock of three bacterial species—Bacillus subtilis, Pseudomonas putida, and Clostridium acetobutylicum—potentially applicable in microbial-enhanced oil recovery processes. The transport of cells during the injection of bacterial suspension and nutrient medium was simulated by a deep bed filtration model. Deep bed filtration coefficients and the maximum capacity of cells in porous rock were measured. Low to intermediate (~10⁶/ml) injection concentrations of cellular suspensions are recommended because plugging of inlet surface is less likely to occur. In addition to their resistance to adverse environments, spores of clostridia are strongly recommended for use in microbial-enhanced oil recovery processes since they are easiest among the species tested to push through porous rock. After injection, further transport of bacteria during incubation can occur by growth and mobility through the stagnant nutrient medium which fills the porous rock. We have developed an apparatus to study the migration of bacteria through a Berea sandstone core containing nutrient medium.     

Role of Microcystis aeruginosa passing through the digestive tracts of filter-feeding animals in eutrophic water reservoirs (review)

The foreign and Russian literature devoted to studying the effect of enhancing the growth of colonies of the cyanobacterium Microcystis aeruginosa Kutz em. Elenk. after their passage in a viable state through the digestive tract of filter-feeding aquatic animals (planktivorous fish, daphnia, and bivalves) has been analyzed. The main mechanisms of this effect are considered. Its role in the functioning of eutrophic reservoirs is discussed. The prospects and the need for further studies of the effect of enhancing Microcyctis growth after its viable passage through the digestive tracts of filter-feeding animals are shown for the development of a complete theory of the functioning of aquatic ecosystems.     

Survival of Yogurt Bacteria in the Human Gut

Whether Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus can be recovered after passage through the human gut was tested by feeding 20 healthy volunteers commercial yogurt. Yogurt bacteria were found in human feces, suggesting that they can survive transit in the gastrointestinal tract.     

Does passage time through the lizard Podarcis lilfordi’s guts affect germination performance in the plant Withania frutescens?

I tested predictions of the hypothesis that a longer seed passage time through the gut of the lizard Podarcis lilfordi enhances germination performance in the plant Withania frutescens. I compared germination success and germination time between seeds that were ingested by lizards and control seeds that were not. I also explored relationships between natural variation in seed passage time and germination performance. Germination success did not differ between ingested (63 %) and non-ingested seeds (56 %); there was no significant relationship between germinability and retention time. Germination time did not differ between ingested and non-ingested seeds, and was unrelated to retention time. Hence, I found no support for the hypothesis that prolonged retention times should improve germination performance. In addition, this is a pilot study stressing the importance of reptiles as seed dispersers, at least in certain habitats.     

A Ligand Channel through the G Protein Coupled Receptor Opsin

The G protein coupled receptor rhodopsin contains a pocket within its seven-transmembrane helix (TM) structure, which bears the inactivating 11-cis-retinal bound by a protonated Schiff-base to Lys296 in TM7. Light-induced 11-cis-/all-trans-isomerization leads to the Schiff-base deprotonated active Meta II intermediate. With Meta II decay, the Schiff-base bond is hydrolyzed, all-trans-retinal is released from the pocket, and the apoprotein opsin reloaded with new 11-cis-retinal. The crystal structure of opsin in its active Ops* conformation provides the basis for computational modeling of retinal release and uptake. The ligand-free 7TM bundle of opsin opens into the hydrophobic membrane layer through openings A (between TM1 and 7), and B (between TM5 and 6), respectively. Using skeleton search and molecular docking, we find a continuous channel through the protein that connects these two openings and comprises in its central part the retinal binding pocket. The channel traverses the receptor over a distance of ca. 70 Å and is between 11.6 and 3.2 Å wide. Both openings are lined with aromatic residues, while the central part is highly polar. Four constrictions within the channel are so narrow that they must stretch to allow passage of the retinal β-ionone-ring. Constrictions are at openings A and B, respectively, and at Trp265 and Lys296 within the retinal pocket. The lysine enforces a 90° elbow-like kink in the channel which limits retinal passage. With a favorable Lys side chain conformation, 11-cis-retinal can take the turn, whereas passage of the all-trans isomer would require more global conformational changes. We discuss possible scenarios for the uptake of 11-cis- and release of all-trans-retinal. If the uptake gate of 11-cis-retinal is assigned to opening B, all-trans is likely to leave through the same gate. The unidirectional passage proposed previously requires uptake of 11-cis-retinal through A and release of photolyzed all-trans-retinal through B.     

Epizootiology of a nuclear polyhedrosis virus in European spruce sawfly (Gilpinia hercyniae): the rate of passage of infective virus through the gut of birds during cage tests.

The passage of a nuclear polyhedrosis virus (NPV) of the sawfly, Gilpinia hercyniae, through avian gut was studied during cage tests on Sturnus vulgaris (three individuals), Parus ater (one), Parus caerulus (five), and Parus major (one). Following brief infection feeds, polyhedral inclusion bodies of the virus could be detected in bird feces within 0.5 hr. Peak passage of polyhedra occurred in less than 1 hr and none were detected after 2.5 hr. The feces of all birds remained infective (in bioassay tests using first instar G. hercyniae larvae) to the end of the day of infection while those of nine birds remained infective to the next day and of six birds to the third day. One bird, P. major, was also infective on Days 4, 6, and 7. The infectivity of NPV in feces stored for 2 years at +3°C declined by half. Though the scale of their epizootiological contribution is unknown, the comparatively long retention and passage of infective virus suggests birds may be effective in short- and long-distance transport of baculoviruses.     

Some aspects of the feeding and behavior of larvae of the Barents Sea capelin Mallotus villosus villosus (Salmoniformes, Osmeridae) in experimental conditions

The results of experimental investigations on the feeding and behavior of larvae of the Barents Sea capelin Mallotus villosus villosus during their transition to exogenous feeding are presented. Data concerning the passage duration of nauplii of Artemia salina along the intestine, feeding intensity, the portion of feeding larvae, swimming speed, sinking speed of non-feeding individuals, distances of responses and escape, as well as certain features of the passage of food through the intestines are given.     

Potential for dissemination of Phytophthora cinnamomi by feral pigs via ingestion of infected plant material

Feral pigs have long been implicated as potential vectors in the spread of the devastating plant pathogen Phytophthora cinnamomi due to their rooting and wallowing activities which may predispose them as vectors of infested soil. In this study, we aim to determine whether feral pigs have the potential to act as vectors of plant pathogens such as P. cinnamomi through their feeding activity. The typically omnivorous diet of feral pigs may also lead to the passage of P. cinnamomi infected plant material through their digestive system. This study investigates the potential for feral pigs to pass viable P. cinnamomi in their faeces following the ingestion of millet seeds, pine plugs and Banksia leptophilia roots inoculated with P. cinnamomi. Recovery rates of P. cinnamomi from the millet seeds, pine plugs and B. leptophilia roots following a single ingested bolus were 33.2, 94.9 and 10.4 %, respectively supported by quantitative PCR analysis. These results demonstrate that P. cinnamomi remain viable within infected plant material following passage through the pig digestive tract, although the digestive processes reduce the pathogen’s viability. An inverse relationship was observed between the viability of infected material and passage time, suggesting that partially digested plant material provides protection for P. cinnamomi against the adverse environmental conditions of the pig digestive tract. Phytophthora cinnamomi remained viable for up to 7 days in larger pieces of colonised woody plant material such as the pine plugs. A plant infection trial using passaged P. cinnamomi colonised pine plugs showed that even material that remained in the digestive tract for 7 days was capable of infecting and killing healthy plants, susceptible to P. cinnamomi. This study provides compelling evidence that feral pigs have the ability to transport viable P. cinnamomi in their digestive tract.     

Predatory behaviour of juvenile shore crab Carcinus maenas (L.)

Juvenile shore crabs Carcinus maenas (L.) were observed feeding on rock barnacles Semibalanus balanoides (L.) on a Bay of Fundy rocky shore. This previously unreported predatory behaviour was further investigated in the laboratory. When given a choice of three common and abundant gastropods, Nucella lapillus (L.), Littorina littorea (L.), and Littorina obtusata (L.), and the rock barnacle Semibalanus balanoides, juvenile shore crabs of both sexes ate mainly barnacles and consumed proportionately more barnacles than gastropods compared with adults, which ate mainly gastropods. The rock barnacle is an abundant and readily available food source which may be important in sustaining the juvenile crab through periods of moults and rapid growth. As the shore crab attains a certain age (size), it must forage lower on the shore as gastropods become more important in its diet.     

Contribution to the problem of mycoplasma-like organism transmigration in plants

Anatomical-histological investigations of slide series revealed that the secondary vascular formations appearing scattered in callus tissue culture derived from a stem piece containing mycoplasma-like organisms (MLO) (potato witches’ broom agent) have no connection with the vascular tissues of the original stem piece. 77% of reconstituted tobacco plants (Nicotiana glauca Grah.) from these callus cultures were infected with MLO. When diseasedNicotiana glauca scions are grafted on healthy tomato plants as stocks, MLO transmigration into the stocks is realized in all cases ten days after grafting; at that time, the scions and stocks are united only by a zone of a few layers of parenchymatous cells without any indication of vascular tissue union between the two components. In both cases, the MLO transmigration could not be realized by passage through sieve pores in sieve tubes. Our results bring indirect evidence of MLO transmigration through parenchymatous cells and thus support the hypothesis of MLO passage through the plasmodesmata.     

Antigen-induced in vitro lymphocyte blastogenesis in the rabbit: a T-cell response.

In vitro lymphocyte stimulation of sensitized rabbit lymphocytes to specific antigen (ovalbumin) was found to depend on thymic-dependent lymphocytes. This conclusion is based on an enhanced response upon enrichment with T lymphocytes by passage of lymphocytes through nylon wool, and on the elimination of the response after treatment of lymphocytes with complement and an antiserum to rabbit thymus cells prepared in a goat. Specificity of the antiserum was demonstrated by elimination of in vitro T-cell function and retention of in vitro B-cell functions.     

Experimental Transmission of Enterobacteriaceae by Insects. I. Fate of Salmonella Fed to the Hide Beetle Dermestes maculatus and a Novel Method for Mounting Insects

A method of mounting insects was devised. The procedure is simple to perform and facilitates quantitative bacteriological studies of feces with a minimal possibility of cross contamination. By this method, it was observed that approximately 10⁷ cells of Salmonella were required for passage through the intestinal tract. Multiple doses of this magnitude were necessary to initiate intestinal infection. The numerical considerations cast doubt that Dermestes is involved significantly in the dissemination of Salmonella in the environment of food and feed plants.