DISTRIBUTION AND MOVEMENTS OF BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS) ALONG THE NORTHERN SOUTH CAROLINA COAST: A RESEARCH PARTNERSHIP WITH ECOTOURISM

Assessments of bottlenose dolphin stocks rely on current information about the distribution and abundance of specific populations. We partnered with two ecotourism dolphin watch businesses to investigate bottlenose dolphins along the northern South Carolina coast. The temporal and spatial distribution of dolphins was examined by comparing dorsal fin images from research-based surveys with photo-identification efforts from commercial dolphin watch groups. Matches between survey locations and times revealed patterns of seasonal residency, potential migratory dispersal between seasons, and variable patterns for home range size. An enhanced understanding of the structure of local populations will contribute to the management of local fisheries, ecotour operations, and coastal ocean health

Characterization of an influenza a host range mutant

A mixed infection of primary chick kidney cells at 38[deg] with A/Ann Arbor/6/60 cold adapted virus and A/Alaska/6/77 wt virus yielded a cold-reassortant virus, CR43-clone 3, which had a host range different from that of either parent. It does not produce detectable virus when grown in Madin-Darby canine kidney cells, while growing normally in primary chick kidney cells at 33[deg]. Both parents, however, grow well in either cell type at 33[deg]. Genotypic analysis of viral RNA electrophoresed in polyacrylamide gels has shown that CR43-clone 3 virus has an aberrant NS gene different from the NS gene of either parent virus. Reassortant viruses made between CR43-clone 3 virus and A/California/ 10/78 (H1N1) virus in primary chick kidney cells at 33[deg] showed the same host range restriction only if the NS gene was derived from the CR43-clone 3 virus. A mixed infection with these same parents, but in Madin-Darby canine kidney cells at 33[deg], produced reassortants that always contained the A/California/10/78 NS gene instead of the CR43clone 3 NS gene. Ferrets inoculated intranasally with the CR43-clone 3 reassortant do not become sick or infected, based on the lack of symptoms: no rhinitis, coryza, or fever; and no detectable virus recovered from nasopharyngeal swabs, turbinate, or lung tissues at 48 hr after infection. Thus, CR43-clone 3 virus contains an aberrant NS gene and manifests a restricted host range phenotype in Madin-Darby canine kidney cells and ferrets.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25081/1/0000512.pd

Development and characterization of cold-adapted viruses for use as live virus vaccines

Representative viruses from twelve RNA and two DNA virus genera have been successfully adapted to growth at sub-optimal temperature (cold-adapted). In almost every case, there was a correlation between acquisition of the cold-adaptation phenotype and loss of virulence in the normal host whether animal or man. Overall, the best method of cold adaptation to develop a live virus vaccine line appeared to be a stepwise lowering of the growth temperature allowing time for multiple lesions to occur and/or be selected. In addition, the starting virus should be a recent isolate not as yet adapted to a tissue culture host and the cold-adaptation process should then occur in a host heterologous to the virus' normal host. These viruses have been reviewed in the light of their cold-adaptation method and successful production of an attenuated line as virus vaccine candidate. Finally, detailed information is presented for the cold-adaptation process in influenza virus.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25479/1/0000019.pd

Terrestrial groundwater and nutrient discharge along the 240-km-long Aquitanian coast

We collected samples from sea water, runnel water, beach pore waters, water from the unconfined surficial aquifer discharging at the beach face, groundwater, and rainwater from the Aquitanian coast in order to determine the flux of dissolved inorganic nitrogen (DIN), phosphorus and silica from terrestrial submarine groundwater discharge (SGD). The flux of fresh groundwater was obtained from a water balance calculation based on precipitation and evapotranspiration and assessment of the coastal watershed from hydrograph separation. Waters with intermediate salinities between sea water and freshwaters are found all along the 240-km-long coast, indicating that SGD is ubiquitous. The estimated fresh water flux is 2.25 m3 d− 1 m− 1 longshore. Terrestrial SGD provides a DIN flux of 9·106 mol each year to the adjacent coastal zone. This flux is about four times lower than the release of DIN due to tidally driven saline SGD. The freshwater DIN flux is low because the upland land use consists almost exclusively of pine forest. Dissolved organic nitrogen represents more than 60% of the total dissolved nitrogen flux. Dissolved iron, phosphorus and silica have much higher concentrations in the anoxic forest aquifer than in the fresh-water end-member of the subterranean estuary sampled in the upper beach aquifer. This suggests that the salinity gradient of the estuary does not correspond to a redox gradient. The redox front between anoxic groundwater and fresh oxic waters occurs below the soil-depleted foredune/yellow dune. Anoxic P- and Si-rich waters seep directly on the beach face only in the north Gironde, where the foredunes are eroded. This study reveals the role of the sandy foredune aquifer in biogeochemical fluxes from SGD, which is to dilute and oxidize waters from the unconfined surficial upland aquifer

Genetics of cold-adapted B/Ann Arbor/1/66 influenza virus reassortants: the acidic polymerase (PA) protein gene confers temperature sensitivity and attenuated virulence

The cold-adapted B/Ann Arbor/1/66 influenza virus (ca B/AA/1/66) expresses temperature-sensitive (ts), cold-adapted (ca) and attenuation phenotypes. Reassortants which inherit one or more genes from ca B/AA/1/66 and all other genes from a virulent, wild-type influenza virus, B/Houston/1732/76, were produced and evaluated in order to identify the gene(s) responsible for the ts, ca and attenuation phenotypes. Only reassortants which inherited the PA gene from ca B/AA/1/66 expressed the ts phenotype in MDCK cells at 39 [deg]C. None of the reassortants tested expressed the ca phenotype in embryonated eggs at 25 [deg]C. The virulence of several reassortants was evaluated in ferrets. Inheritance of the PA gene from ca B/AA/1/66 was correlated with significant febrile attentuation and the apparent restriction of viral replication in the lower respiratory tract. Isolation of a virulent, non-ts revertant virus inheriting only the PA gene from ca B/AA/1/66 established a direct relationship between expression of the ts phenotype and attenuated virulence. Evidence for the contribution of at least one other gene from ca B/AA/1/66 to attenuation was observed. Thus, based on the methods used to determine reassortant gene compositions, these results indicate that the PA gene is primarily responsible for attenuation of ca B/AA/1/66 and its reassortants.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26616/1/0000157.pd

Virus occurrence and transport in a school septic system and unconfined aquifer

A study was conducted to analyze viral indicators of fecal contamination using a high school population as a community source. The study also examined the occurrence, distribution and transport rate of viruses in a productive sand and gravel aquifer. Concentrated septic tank effluent samples were tested for virucidal activity. Results indicated that recovery values for enteroviruses in the ground water ranged between 5 and 30%

Cold-adapted reassortants of influenza A virus: Pathogenicity of A/Ann Arbor/6/60×A/Alaska/6/77 reassortant viruses In vivo and In vitro

Cold-adapted reassortants of A/Ann Arbor/6/60×A/Alaska/6/77 viruses made in MDCK cells have recently been assessed genotypically and for temperature-sensitive and cold-adapted phenotypes. These reassortants were used to infect ferrets and hamsters and to inoculate organ cultures of hamster tracheal rings, in order to assess their degree of virulence. Virulence in the three model systems corresponded quite well, and a correlation between loss of virulence and particular A/AA/6/60 genes present in the reassortants was noted. Two different reassortants containing either RNA 2 or RNA 5 (NA gene) alone from A/AA/6/60 showed little attenuation from the wild-type parent. A reassortant containing both RNA2 and the NA gene from A/AA/6/60 and all remaining wild-type genes showed some small decrease in virulence compared to the wild-type virus. However a reassortant containing these two A/AA/6/60 genes and RNA 3 as an additional gene from this parent, had a level of attenuation comparable to that of the cold-adapted virus.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41693/1/705_2005_Article_BF01316727.pd

Resolution of a common RNA sequencing ambiguity by terminal deoxynucleotidyl transferase

One of the more common ambiguities which arise when using reverse transcriptase and dideoxynucleotide-chain termination to sequence RNA is a radioactive band of cDNA that extends over all four lanes on a sequencing gel. The adjacent sequences both above and below the band are not affected. Assuming then, that these ambiguities are caused by the termination of the DNA polymerase activity of reverse transcriptase for reasons other than the insertion of a dideoxynucleotide in the growing cDNA chain, terminal deoxynucleotidyl transferase should be able to continue to add deoxynucleotides to these products after the sequencing reaction is complete. It does, clearing the improperly terminated cDNA from these pileup sites, revealing the correct sequence. This technique can also be used to identify the template RNA's 5'-terminal base, although far more units of terminal deoxynucleotidyl transferase are required.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26058/1/0000132.pd

A mutation in the PA protein gene of cold-adapted B/Ann Arbor/1/66 influenza virus associated with reversion of temperature sensitivity and attenuated virulence

Reassortant SG3 inherits only the acidic polymerase (PA) protein gene from the cold-adapted B/AA/1/66 influenza virus (ca B/AA/l/66) and all remaining genes from a virulent, wild-type virus. This reassortant demonstrates attenuated virulence in ferrets and expresses a is phenotype characteristic of the ca parent. During virulence evaluation of SG3, a virulent, non-ts revertant virus (designated SG3rFL) was isolated from the lungs of one ferret. In order to determine whether the reversion of SG3 resulted from mutation of the PA gene and/or as the result of extragenic supressor mutations, the revertant PA gene of SG3rFL was transferred to a reassortant (SG3r) inheriting only the revertant PA gene from SG3rFL and all remaining genes from SG3. Reassortant SG3r was non-ts and virulent, indicating that mutation of the PA gene was sufficient for the reversion of the is and attenuation phenotypes expressed by SG3rFL. The nucleotide and predicted amino acid sequences of the SG3rFL PA gene were determined and compared to those of wt and ca B/AA/1 /66. The predicted PA proteins of wt and ca B/AA/1 /66 are known to differ by six amino acid substitutions including a valine to methionine substitution at residue 431. The PA proteins of ca B/AA/1/66 and SG3rFL were distinguished by only the single amino acid substitution of methionine to isoluecine also occurring at residue 431. Thus, the methionine residue was implicated in the attenuation of ca B/AA/1/66 and its reassortants. The hydropathic properties of valine, isoleucine, and methionine suggested that reversion involved the restoration of hydrophobic character at this site.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27363/1/0000388.pd

Sequence comparison of wild-type and cold-adapted B/Ann Arbor/1/66 influenza virus genes

Consensus sequences for both wt and ca B/Ann Arbor/1 /66 viral PB2, PB1, PA, NP, M, and NS genes were directly determined from vRNA using a combination of chemical and chain-termination sequencing methods. There were 105 sites of difference between the wt and ca sets of these six RNA genes. The differences resulted in 26 amino acid substitutions distributed over the six proteins. The sequence changes were compared to the sequences of other known influenza type B wt viruses to pinpoint those changes that were unique to the ca B/Ann Arbor/1/66 virus. Of the 26 amino acid differences, only 11 were unique to the cold-adapted virus. These unique sites were distributed among five of the six genes. The NS protein had no amino acid substitutions. The sequence changes are discussed in terms of their probable mode of origin and selection, and in terms of their importance to the cold-adapted, temperature-sensitive, and attenuation phenotypes of ca B/AA/1 /66 virus. The sequence and organization of the PB2 gene and predicted protein are also given. The PB2 gene was 2396 nucleotides long, and it encoded a predicted protein of 770 amino acids with a molecular weight of 88,035 Da for the wt virus and 88,072 Da for the ca virus. Both proteins were predominantly hydrophilic, and each had an overall charge of +24.5 at pH 7.0.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27362/1/0000387.pd