Physical Assay and Growth Cycle Studies of a Defective Adeno-Satellite Virus

Electron microscopic particle counting of the defective adeno-satellite virus (ASV), by use of pseudoreplication and negative staining with phosphotungstic acid, was shown to be a reproducible quantitative assay procedure. Particles of satellite type 4 that were counted in fluids from infected cultures had the same morphology as particles that banded at a buoyant density of 1.43 g/cc in cesium chloride. Other satellite virus serotypes examined in the same manner had a buoyant density of 1.37 to 1.38 g/cc. A comparison of satellite titers obtained by complement fixation and by particle counting demonstrated that an increase in satellite particles resulted in a corresponding increase in CF titers; however, electron microscopy was at least 10 times more sensitive than complement fixation for detecting satellite virus. Growth cycle studies of satellite virus in cells co-infected with adenovirus, as assayed by particle counting, indicated that the kinetics of satellite virus production closely followed the kinetics of its helper adenovirus production, with an eclipse period of 12 to 16 hr. The eclipse period of the satellite remained the same when cultures were preinfected with satellite 24 hr prior to adenovirus inoculation. However, when cultures were infected with adenovirus 12 hr before satellite virus, the eclipse period of the satellite was shortened to between 4 and 6 hr. Thus, satellite virus replication seems dependent upon a relatively late event in the adenovirus replication cycle. When cells were co-infected with adenovirus and its defective satellite, the yield of adenovirus was markedly reduced from that obtained in cells singly infected with adenovirus.     

Tissue culture of Ecklonia radiata (Phaeophyceae,Laminariales): effects on growth of light,organic carbon source and vitamins

The effects of light quality and irradiance, and supply of organic carbon and vitamins on the growth of two forms of Ecklonia radiata in tissue culture were examined. A callus of unpigmented cells developed over the cut surface of newly excised explants of stipe. This growth was best in the dark but stopped after 10 weeks. Pigmented, mainly filamentous clumps of cells developed from explants after several weeks in culture. These required light for growth, with growth being enhanced by increasing photon flux density up to 30 μmol photon m^-2 s^-1, with the active spectral component being red light (> 600 nm). The addition to the medium of a range of organic carbon sources or vitamins did not stimulate growth of either culture type in the dark. author for correspondence     

Growth of food-borne pathogenic bacteria in oil-in-water emulsions: I—Methods for investigating the form of growth

Methods are presented for investigating the site and form of growth of bacteria in model oil-in-water emulsions and in dairy cream. Following growth of the bacteria, the continuous aqueous phase is gelled using agarose and the oil phase removed using a mixture of chloroform and methanol. Using this method, the authors have found that Listeria monocytogenes, Salmonella typhimurium and Yersinia enterocolitica grow in the form of colonies in concentrated oil-in-water emulsions. Colonies of L. monocytogenes and Y. enterocolitica also form in artificially-inoculated fresh and tinned dairy cream. If information about the precise site of growth is not required, the authors have discovered that intact colonies can be liberated from the model emulsions by dissolving away the oil phase with chloroform: methanol.     

Analysis ofDrosophila chromosome4 using pulsed field gel electrophoresis

Previous estimates of the size ofDrosophila melanogaster chromosome4 have indicated that it is 1% to 4% of the genome or 6 Mb. We have used pulsed field gel electrophoresis (PFGE) to separate megabase-sized molecules ofD. melanogaster chromosomal DNA. Southern blots of these gels were probed with DNA fragments from thecubitus interruptus andzfh-2 genes, which are located on chromosome4. They each identify the same-sized distinct band that migrates at approximately 5.2 Mb in DNA preparations from the Kc cell line. We interpret this band to be intact chromosome4. In DNA obtained from embryos of variousD. melanogaster wild-type strains, this chromosome band showed strain-specific size variation that ranged from 4.5 to 5.2 Mb. TheD. melanogaster chromosome4 probes also identified a single, 2.4 Mb band in embryonic DNA fromDrosophila simulans. We conclude thatD. simulans chromosome4 is substantially smaller than that ofD. melanogaster, presumably owing to diffirences in the amount of heterochromatic DNA sequences. Our simple DNA preparation from embryos and PFGE conditions should permit preparative isolation of chromosome4 DNA and will facilitate the molecular mapping of this chromosome.     

Coexisting mangrove-coral habitat use by reef fishes in the Caribbean

We conducted visual fish surveys in coexisting mangrove-coral (CMC) habitats in Panama to analyze the effect of coral presence in mangrove habitats on the fish assemblage. Our study revealed that CMC habitats harbor distinct fish assemblages compared to mangrove habitats without coral, with greater species richness and increased herbivore abundance. Abstract in Spanish is available with online material.     

Old,socially housed rhesus monkeys manipulate objects

Recent research has indicated that old, individually housed monkeys show little interest in novel objects. Yet unanswered is whether this effect is caused primarily by age or housing condition. The purpose of this study was to assess the role of social living in promoting responsiveness to objects. We measured the rates of object manipulation in older animals, assessed responsiveness over time to particular objects as a measure of habituation, and examined social influences on object use. Several social groups of rhesus monkeys that contained older adults were studied. These groups were housed in indoor pens or in an outdoor enclosure, and all monkeys had continuous access to a variety of objects in their home environment. In contrast to previous studies of individually housed monkeys, our group-housed monkeys showed sustained interest in objects. Old monkeys manipulated objects extensively, and this response was all the more significant, given that the objects were not novel. Monkeys housed in an outdoor enclosure showed object manipulation patterns that were not different from monkeys housed in indoor pens. However, females exhibited much higher object-related responses than males. Social facilitation played a role in the reactions of some monkeys to objects. Patterns of social facilitation as well as avoidance were present in two of the three indoor groups that were observed. Failure to manipulate objects in rhesus macaques appears to be more a function of individual housing than of old age. Factors such as environmental complexity, social needs, and early experience should be considered in order to understand why individually housed rhesus monkeys are unresponsive to objects. © 1993 Wiley-Liss, Inc.     

Mammal use of riparian corridors in semi-arid Sonora,Mexico

Wildlife populations in semi-arid regions are increasingly challenged by human activities and dependent on the connectivity of riparian corridors for access to surface water. The Madrean Archipelago is a biodiversity hotspot along the arid United States–Mexico borderlands that support both Neotropical and Nearctic wildlife. Infrastructure development (e.g., the border wall and the expansion of Mexican Federal Highway 2) in this region inhibits wildlife movement along the transnational mountain archipelago by disconnecting habitat. To explore the relationship between habitat variables and mammal use of riparian corridors in northern Sonora, Mexico, we collected data from 19 motion-sensitive cameras between October 2018 and April 2019 and used single-season occupancy models and Royle-Nichols abundance estimation models to analyze our data. We recorded 21 species of mammals, including the first sighting of jaguar (Panthera onca) in this region in 25 years. River characteristics (distance from river, riparian corridor width, water availability), remoteness (distance from highway, productivity, elevation), and topographic variety (vertical elevation difference) influenced patterns of occupancy probability and estimated abundance of mammals >1 kg, but the strength and direction of these relationships varied by species. Additionally, intermittently wet desert washes were comparable in species richness to the perennial system. These results highlight the importance of examining physical and biological aspects of habitat. This is especially true when identifying corridors where mitigation structures should be placed to improve wildlife connectivity in biodiversity hotspots like the Madrean Archipelago and semi-arid ecosystems worldwide.     

The hmu locus of Yersinia pestis is essential for utilization of free haemin and haem-protein complexes as iron sources

Yersinia pestis strains utilize haem and several haem-protein complexes as sole sources of iron. In this study, the haemin uptake locus (hmu) of Y. pestis KIM6+ was selected from a genomic library by trans-duction into an Escherichia coli siderophore synthesis (entC) mutant. Recombinant plasmids containing a common 16 kb BamHI insert were isolated that allowed E. coli entC to use haemin as an iron source. An 8.6 kb region of this insert was found to be essential for haemin utilization and encoded at least five proteins with molecular masses of 79/77, 44, 37, 35, and 30/27.5 kDa. A 10.9 kb Clal fragment containing the hmu locus showed varying degrees of homology to genomic DNA from Yersinia pseudotuberculosis, Yersinia enter-ocolitica, and other genera of Enterobacteriaceae. An E. coli hemA aroB strain harbouring cloned hmu genes used haemin as both an iron and porphyrin source but only on iron-poor medium, suggesting that haemin uptake is tightly iron regulated. Additionally, haemoglobin and myoglobin were used as iron sources by an E. coli entC (pHMU2.2) strain. Deletion of the hmu locus from Y. pestis KIM6+ chromosome generated a mutant that grew poorly on iron-depleted medium containing free haemin as well as mammalian haem-protein complexes including haemoglobin, haemoglobin-haptoglobin, myoglobin, haem-haemopexin, and haem-albumin unless it was complemented with cloned hmu genes.