Proceedings of the International Workshop on age determination of oceanic pelagic fishes: Tunas, billfishes, and sharks, Miami, Florida, February 15-18,1982

Accurate and precise estimates of age and growth rates are essential parameters in understanding the population dynamics of fishes. Some of the more sophisticated stock assessment models, such as virtual population analysis, require age and growth information to partition catch data by age. Stock assessment efforts by regulatory agencies are usually directed at specific fisheries which are being heavily exploited and are suspected of being overfished. Interest in stock assessment of some of the oceanic pelagic fishes (tunas, billfishes, and sharks) has developed only over the last decade, during which exploitation has increased steadily in response to increases in worldwide demand for these resources. Traditionally, estimating the age of fishes has been done by enumerating growth bands on skeletal hardparts, through length frequency analysis, tag and recapture studies, and raising fish in enclosures. However, problems related to determining the age of some of the oceanic pelagic fishes are unique compared with other species. For example, sampling is difficult for these large, highly mobile fishes because of their size, extensive distributions throughout the world's oceans, and for some, such as the marlins, infrequent catches. In addition, movements of oceanic pelagic fishes often transect temperate as well as tropical oceans, making interpretation of growth bands on skeletal hardparts more difficult than with more sedentary temperate species. Many oceanic pelagics are also long-lived, attaining ages in excess of 30 yr, and more often than not, their life cycles do not lend themselves easily to artificial propagation and culture. These factors contribute to the difficulty of determining ages and are generally characteristic of this group-the tunas, billfishes, and sharks. Accordingly, the rapidly growing international concern in managing oceanic pelagic fishes, as well as unique difficulties in ageing these species, prompted us to hold this workshop. Our two major objectives for this workshop are to: I) Encourage the interchange of ideas on this subject, and 2) establish the "state of the art." A total of 65 scientists from 10 states in the continental United States and Hawaii, three provinces in Canada, France, Republic of Senegal, Spain, Mexico, Ivory Coast, and New South Wales (Australia) attended the workshop held at the Southeast Fisheries Center, Miami, Fla., 15-18 February 1982. Our first objective, encouraging the interchange of ideas, is well illustrated in the summaries of the Round Table Discussions and in the Glossary, which defines terms used in this volume. The majority of the workshop participants agreed that the lack of validation of age estimates and the means to accomplish the same are serious problems preventing advancements in assessing the age and growth of fishes, particularly oceanic pelagics. The alternatives relating to the validation problem were exhaustively reviewed during the Round Table Discussions and are a major highlight of this workshop. How well we accomplished our second objective, to establish the "state of the art" on age determination of oceanic pelagic fishes, will probably best be judged on the basis of these proceedings and whether future research efforts are directed at the problem areas we have identified. In order to produce high-quality papers, workshop participants served as referees for the manuscripts published in this volume. Several papers given orally at the workshop, and included in these proceedings, were summarized from full-length manuscripts, which have been submitted to or published in other scientific outlets-these papers are designated as SUMMARY PAPERS. In addition, the SUMMARY PAPER designation was also assigned to workshop papers that represented very preliminary or initial stages of research, cursory progress reports, papers that were data shy, or provide only brief reviews on general topics. Bilingual abstracts were included for all papers that required translation. We gratefully acknowledge the support of everyone involved in this workshop. Funding was provided by the Southeast Fisheries Center, and Jack C. Javech did the scientific illustrations appearing on the cover, between major sections, and in the Glossary. (PDF file contains 228 pages.)

Management of northern fur seals on the Pribilof Islands, Alaska, 1786-1981

This paper includes information about the Pribilof Islands since their discovery by Russia in 1786 and the population of northern fur seals, Cailorhinus ursinus, that return there each summer to bear young and to breed. Russia exterminated the native population of sea Oilers, Enhydra lulris, here and nearly subjected the northern fur seal to the same fate before providing proper protection. The northern fur seal was twice more exposed to extinction following the purchase of Alaska and the Pribilof Islands by the United States in 1867. Excessive harvesting was stopped as a result of strict management by the United States of the animals while on land and a treaty between Japan, Russia, Great Britain (for Canada), and the United States that provided needed protection at sea. In 1941, Japan abrogated this treaty which was replaced by a provisional agreement between Canada and the United States that protected the fur seals in the eastern North Pacific Ocean. Japan, the U.S.S.R., Canada, and the United States again insured the survival of these animals with ratification in 1957 of the "Interim Convention on the Conservation of North Pacific Fur Seals," which is still in force. Under the auspices of this Convention, the United States launched an unprecedented manipulation of the resource through controlled removal during 1956-68 of over 300,000 females considered surplus. The biological rationale for the reduction was that production of fewer pups would result in a higher pregnancy rate and increased survival, which would, in turn, produce a sustained annual harvest of 55,000-60,000 males and 10,000-30,000 females. Predicted results did not occur. The herd reduction program instead coincided with the beginning of a decline in the number of males available for harvest. Suspected but unproven causes were changes in the toll normally accounted for by predation, disease, adverse weather, and hookworms. Depletion of the animals' food supply by foreign fishing Heets and the entanglement of fur seals in trawl webbing and other debris discarded at sea became a prime suspect in altering the average annual harvest of males on the Pribilof Islands from 71,500 (1940-56) to 40,000 (1957-59) to 36,000 (1960) to 82,000 (1961) and to 27,347 (1972-81). Thus was born the concept of a research control area for fur seals, which was agreed upon by members of the Convention in 1973 and instituted by the United States on St. George Island beginning in 1974. All commercial harvesting of fur seals was stopped on St. George Island and intensive behavioral studies were begun on the now unharvested population as it responds to the moratorium and attempts to reach its natural ceiling. The results of these and other studies here and on St. Paul Island are expected to eventually permit a comparison between the dynamics of unharvested and harvested populations, which should in turn permit more precise management of fur seals as nations continue to exploit the marine resources of the North Pacific Ocean and Bering Sea. (PDF file contains 32 pages.)

Proceedings of the Twenty-third Annual Symposium on sea turtle biology and conservation, 17 to 21 March 2003, Kuala Lumpur, Malaysia

The 23rd Annual Symposium on Sea Turtle Biology and Conservation was held between 17 and 21 March 2003 at The Legend Hotel in Kuala Lumpur, Malaysia, hosted by the Community Conservation Network, Hawaii, and WWF-Malaysia. The meeting was attended by slightly more than 300 participants representing 73 countries, a dramatic drop in participation from previous years brought about in no small part by the looming war in the middle east region and concerns over travel safety. For 22 years the Symposium had bee an Americas-based event, even though it is the annual gathering of the "international" sea turtle society, and with the move to Malaysia, the Symposium hoped to raise the awareness among the general public of the plight of amrine turtles in Southeast Asia, and share the enormous exspertise of the world authorities on sea turtles with this so-far underrepresented region. Adopting the thems, "Living With Turtles", the Symposium had a very personal flavour, and the smaller number of participants made it possible to make and renew acquaintances, and have time for discussion between sessions. While the travel safety concern excuse was often quoted, it was a pity, particularly to the large contingent of people who attended the event for the first time from underrepresented regions, that many of the household names linked to marine turtle biology and conservation were not present to share their knowledge and promote the global concerns on the plight of turtle populations.

Underwater Passive Acoustic Monitoring for Remote Regions, Hawaii Institute of Marine Biology, Coconut Island, Hawaii, February 7-9, 2007: workshop proceedings

The Alliance for Coastal Technologies (ACT) convened a workshop, sponsored by the Hawaii-Pacific and Alaska Regional Partners, entitled Underwater Passive Acoustic Monitoring for Remote Regions at the Hawaii Institute of Marine Biology from February 7-9, 2007. The workshop was designed to summarize existing passive acoustic technologies and their uses, as well as to make strategic recommendations for future development and collaborative programs that use passive acoustic tools for scientific investigation and resource management. The workshop was attended by 29 people representing three sectors: research scientists, resource managers, and technology developers. The majority of passive acoustic tools are being developed by individual scientists for specific applications and few tools are available commercially. Most scientists are developing hydrophone-based systems to listen for species-specific information on fish or cetaceans; a few scientists are listening for biological indicators of ecosystem health. Resource managers are interested in passive acoustics primarily for vessel detection in remote protected areas and secondarily to obtain biological and ecological information. The military has been monitoring with hydrophones for decades;however, data and signal processing software has not been readily available to the scientific community, and future collaboration is greatly needed. The challenges that impede future development of passive acoustics are surmountable with greater collaboration. Hardware exists and is accessible; the limits are in the software and in the interpretation of sounds and their correlation with ecological events. Collaboration with the military and the private companies it contracts will assist scientists and managers with obtaining and developing software and data analysis tools. Collaborative proposals among scientists to receive larger pools of money for exploratory acoustic science will further develop the ability to correlate noise with ecological activities. The existing technologies and data analysis are adequate to meet resource managers' needs for vessel detection. However, collaboration is needed among resource managers to prepare large-scale programs that include centralized processing in an effort to address the lack of local capacity within management agencies to analyze and interpret the data. Workshop participants suggested that ACT might facilitate such collaborations through its website and by providing recommendations to key agencies and programs, such as DOD, NOAA, and I00s. There is a need to standardize data formats and archive acoustic environmental data at the national and international levels. Specifically, there is a need for local training and primers for public education, as well as by pilot demonstration projects, perhaps in conjunction with National Marine Sanctuaries. Passive acoustic technologies should be implemented immediately to address vessel monitoring needs. Ecological and health monitoring applications should be developed as vessel monitoring programs provide additional data and opportunities for more exploratory research. Passive acoustic monitoring should also be correlated with water quality monitoring to ease integration into long-term monitoring programs, such as the ocean observing systems. [PDF contains 52 pages]

Measures of turbidity in coastal waters, Coconut Island, Oahu, Hawaii, August 31 - September 2, 2005: workshop proceedings

A three day workshop on turbidity measurements was held at the Hawaii Institute of Marine Biology from August 3 1 to September 2, 2005. The workshop was attended by 30 participants from industry, coastal management agencies, and academic institutions. All groups recognized common issues regarding the definition of turbidity, limitations of consistent calibration, and the large variety of instrumentation that nominally measure "turbidity." The major recommendations, in order of importance for the coastal monitoring community are listed below: 1. The community of users in coastal ecosystems should tighten instrument design configurations to minimize inter-instrument variability, choosing a set of specifications that are best suited for coastal waters. The IS0 7027 design standard is not tight enough. Advice on these design criteria should be solicited through the ASTM as well as Federal and State regulatory agencies representing the majority of turbidity sensor end users. Parties interested in making turbidity measurements in coastal waters should develop design specifications for these water types rather than relying on design standards made for the analysis of drinking water. 2. The coastal observing groups should assemble a community database relating output of specific sensors to different environmental parameters, so that the entire community of users can benefit from shared information. This would include an unbiased, parallel study of different turbidity sensors, employing a variety of designs and configuration in the broadest range of coastal environments. 3. Turbidity should be used as a measure of relative change in water quality rather than an absolute measure of water quality. Thus, this is a recommendation for managers to develop their own local calibrations. See next recommendation. 4. If the end user specifically wants to use a turbidity sensor to measure a specific water quality parameter such as suspended particle concentration, then direct measurement of that water quality parameter is necessary to correlate with 'turbidity1 for a particular environment. These correlations, however, will be specific to the environment in which they are measured. This works because there are many environments in which water composition is relatively stable but varies in magnitude or concentration. (pdf contains 22 pages)

In situ measurement of dissolved inorganic carbon speciation in natural waters, pH, pC02, TA, & TC02, Honolulu, Hawaii, February 16-18, 2005: workshop proceedings

The Alliance for Coastal Technology (ACT) convened a workshop on the in situ measurement of dissolved inorganic carbon species in natural waters in Honolulu, Hawaii, on February 16, 17, and 18, 2005. The workshop was designed to summarize existing technologies for measuring the abundance and speciation of dissolved inorganic carbon and to make strategic recommendations for future development and application of these technologies to coastal research and management. The workshop was not focused on any specific technology, however, most of the attention of the workshop was on in situ pC02 sensors given their recent development and use on moorings for the measurement of global carbon fluxes. In addition, the problems and limitations arising from the long-term deployment of systems designed for the measurement of pH, total dissolved inorganic carbon (DIC), and total alkalinity (TA) were discussed. Participants included researchers involved in carbon biogeochemistry, industry representatives, and coastal resource managers. The primary questions asked during the workshop were: I. What are the major impediments to transform presently used shipboard pC02 measurement systems for use on cost-eficient moorings? 2. What are the major technical hurdles for the in situ measurement of TA and DIC? 3. What specific information do we need to coordinate efforts for proof of concept' testing of existing and new technologies, inter-calibration of those technologies, better software development, and more precise knowledge quantzjjing the geochemistry of dissolved inoeanic carbon species in order to develop an observing system for dissolved inorganic carbon? Based on the discussion resulting from these three questions, the following statements were made: Statement No. 1 Cost-effective, self-contained technologies for making long-term, accurate measurements of the partial pressure of C02 gas in water already exist and at present are ready for deployment on moorings in coastal observing systems. Statement No. 2 Cost-effective, self-contained systems for the measurement of pH, TA, and DIC are still needed to both fully define the carbonate chemistry of coastal waters and the fluxes of carbon between major biogeochemical compartments (e.g., air-sea, shelf-slope, water column-sediment, etc.). (pdf contains 23 pages)

Direct numerical simulation of hypersonic boundary layer transition over a blunt cone with a small angle of attack

The direct numerical simulation of boundary layer transition over a 5° half-cone-angle blunt cone is performed. The free-stream Mach number is 6 and the angle of attack is 1°. Random wall blow-and-suction perturbations are used to trigger the transition. Different from the authors’ previous work [Li et al., AIAA J. 46, 2899(2008)], the whole boundary layer flow over the cone is simulated (while in the author’s previous work, only two 45° regions around the leeward and the windward sections are simulated). The transition location on the cone surface is determined through the rapid increase in skin fraction coefficient (Cf). The transition line on the cone surface shows a nonmonotonic curve and the transition is delayed in the range of 0° ≤ θ ≤ 30° (θ = 0° is the leeward section). The mechanism of the delayed transition is studied by using joint frequency spectrum analysis and linear stability theory (LST). It is shown that the growth rates of unstable waves of the second mode are suppressed in the range of 20° ≤ θ ≤ 30°, which leads to the delayed transition location. Very low frequency waves VLFWs� are found in the time series recorded just before the transition location, and the periodic times of VLFWs are about one order larger than those of ordinary Mack second mode waves. Band-pass filter is used to analyze the low frequency waves, and they are deemed as the effect of large scale nonlinear perturbations triggered by LST waves when they are strong enough.The direct numerical simulation of boundary layer transition over a 5° half-cone-angle blunt cone is performed. The free-stream Mach number is 6 and the angle of attack is 1°. Random wall blow-and-suction perturbations are used to trigger the transition. Different from the authors’ previous work [ Li et al., AIAA J. 46, 2899 (2008) ], the whole boundary layer flow over the cone is simulated (while in the author’s previous work, only two 45° regions around the leeward and the windward sections are simulated). The transition location on the cone surface is determined through the rapid increase in skin fraction coefficient (Cf). The transition line on the cone surface shows a nonmonotonic curve and the transition is delayed in the range of 20° ≤ θ ≤ 30° (θ = 0° is the leeward section). The mechanism of the delayed transition is studied by using joint frequency spectrum analysis and linear stability theory (LST). It is shown that the growth rates of unstable waves of the second mode are suppressed in the range of 20° ≤ θ ≤ 30°, which leads to the delayed transition location. Very low frequency waves (VLFWs) are found in the time series recorded just before the transition location, and the periodic times of VLFWs are about one order larger than those of ordinary Mack second mode waves. Band-pass filter is used to analyze the low frequency waves, and they are deemed as the effect of large scale nonlinear perturbations triggered by LST waves when they are strong enough.

A preliminary investigation into the effect of different storage methods on the keeping quality of smoked Oreochromis niloticus

The study was carried out to asses the nutritional qualities of smoked O. niloticus and to discover the best methods of storage to minimize spoilage and infestation of smoked fish. Result showed that the protein contents in A and D decreased while the protein contents of b and C increased. The lipid content increased only in A while it decreased in B-C and D. The moisture content generally increased over the period of storage and there was an increase in ash content only in C while it decreased in A, B and D. The samples packed in polythene bag suffered about 35% mould infection and a few were attached by rodents with some fouling. Samples packed in jute bag were in good condition but were slightly attached by insect. All samples packed in carton and basket were still in good state but there were insect attack in those packed in carton

Climate change adaptation and planning: An example from Kailua Beach, Oahu, Hawaii

The University of Hawaii Sea Grant College Program (UHSG) in partnership with the Hawaii Department of Land and Natural Resources (DLNR), Office of Conservation and Coastal Lands (OCCL) is developing a beach and dune management plan for Kailua Beach on the eastern shoreline of Oahu. The objective of the plan is to develop a comprehensive beach management and land use development plan for Kailua Beach that reflects the state of scientific understanding of beach processes in Kailua Bay and abutting shoreline areas and is intended to provide long-term recommendations to adapting to climate change including potential coastal hazards such as sea level rise. The development of the plan has lead to wider recognition of the significance of projected sea level rise to the region and provides the rational behind some of the land use conservation strategies. The plan takes on a critical light given global predictions for continued, possibly accelerated, sea-level rise and the ongoing focus of intense development along the Hawaiian shoreline. Hawaii’s coastal resource managers are faced with the daunting prospect of managing the effects of erosion while simultaneously monitoring and regulating high-risk coastal development that often impacts the shoreline. The beach and dune preservation plan is the first step in a more comprehensive effort prepare for and adapt to sea level rise and ensure the preservation of the beach and dune ecosystem for the benefit of present and future generations. The Kailua Beach and Dune Management plan is intended to be the first in a series of regional plans in Hawaii to address climate change adaptation through land use planning. (PDF contains 3 pages)

Sea-level rise in Hawaii: Implications for future shoreline locations and Hawaii coastal management

Management of coastal development in Hawaii is based on the location of the certified shoreline, which is representative of the upper limit of marine inundation within the last several years. Though the certified shoreline location is significantly more variable than long-term erosion indicators, its migration will still follow the coastline's general trend. The long-term migration of Hawaii’s coasts will be significantly controlled by rising sea level. However, land use decisions adjacent to the shoreline and the shape and nature of the nearshore environment are also important controls to coastal migration. Though each of the islands has experienced local sea-level rise over the course of the last century, there are still locations across the islands of Kauai, Oahu, and Maui, which show long- term accretion or anomalously high erosion rates relative to their regions. As a result, engineering rules of thumb such as the Brunn rule do not always predict coastal migration and beach profile equilibrium in Hawaii. With coastlines facing all points of the compass rose, anthropogenic alteration of the coasts, complex coastal environments such as coral reefs, and the limited capacity to predict coastal change, Hawaii will require a more robust suite of proactive coastal management policies to weather future changes to its coastline. Continuing to use the current certified shoreline, adopting more stringent coastal setback rules similar to Kauai County, adding realistic sea-level rise components for all types of coastal planning, and developing regional beach management plans are some of the recommended adaptation strategies for Hawaii. (PDF contains 4 pages)

Integrating climate change impacts to improve understanding of coastal climate change: heavy rains, strong winds, and high seas in coastal Hawaii, Alaska and the Pacific Northwest

Coastal storms, and the strong winds, heavy rains, and high seas that accompany them pose a serious threat to the lives and livelihoods of the peoples of the Pacific basin, from the tropics to the high latitudes. To reduce their vulnerability to the economic, social, and environmental risks associated with these phenomena (and correspondingly enhance their resiliency), decision-makers in coastal communities require timely access to accurate information that affords them an opportunity to plan and respond accordingly. This includes information about the potential for coastal flooding, inundation and erosion at time scales ranging from hours to years, as well as the longterm climatological context of this information. The Pacific Storms Climatology Project (PSCP) was formed in 2006 with the intent of improving scientific understanding of patterns and trends of storm frequency and intensity - “storminess”- and related impacts of these extreme events. The project is currently developing a suite of integrated information products that can be used by emergency managers, mitigation planners, government agencies and decision-makers in key sectors, including: water and natural resource management, agriculture and fisheries, transportation and communication, and recreation and tourism. The PSCP is exploring how the climate-related processes that govern extreme storm events are expressed within and between three primary thematic areas: heavy rains, strong winds, and high seas. To address these thematic areas, PSCP has focused on developing analyses of historical climate records collected throughout the Pacific region, and the integration of these climatological analyses with near-real time observations to put recent weather and climate events into a longer-term perspective.(PDF contains 4 pages)

The island of Kauai, Hawaii's progressive shoreline setback and coastal protection ordinance

Approximately two-thirds of coastal and Great Lakes states have some type of shoreline construction setback or construction control line requiring development to be a certain distance from the shoreline or other coastal feature (OCRM, 2008). Nineteen of 30 coastal states currently use erosion rates for new construction close to the shoreline. Seven states established setback distances based on expected years from the shoreline: the remainder specify a fixed setback distance (Heinz Report, 2000). Following public hearings by the County of Kauai Planning Commission and Kauai County Council, the ‘Shoreline Setback and Coastal Protection Ordinance’ was signed by the Mayor of Kauai on January 25, 2008. After a year of experience implementing this progressive, balanced shoreline setback ordinance several amendments were recently incorporated into the Ordinance (#887; Bill #2319 Draft 3). The Kauai Planning Department is presently drafting several more amendments to improve the effectiveness of the Ordinance. The intent of shoreline setbacks is to establish a buffer zone to protect shorefront development from loss due to coastal erosion - for a period of time; to provide protection from storm waves; to allow the natural dynamic cycles of erosion and accretion of beaches and dunes to occur; to maintain beach and dune habitat; and, to maintain lateral beach access and open space for the enjoyment of the natural shoreline environment. In addition, a primary goal of the Kauai setback ordinance is to avoid armoring or hardening of the shore which along eroding coasts has been documented to ultimately eliminate the fronting beach. (PDF contains 4 pages)

Studies on hymenopteran parasitism of Drosophila

Studies on Hymenopteran Parasitism of Drosophila

Flies of the genus Drosophila are subject to attack by a number of parasitic forms. Sturtevant (1921) has listed records of parasitism by protozoa (Leptomonas), fungi (Muiaria and Stigmatomyces), nematodes, mites and v~rious hymenoptera. According to Sturtevant, Perkins (1913) has bred at least five species of hymenoptera, belonging to the proctotrupoid, cynipoid and chalcidoid groups, upon Drosophiline flies. H.S. Smith has bred an unidentified proctotrupoid and a chalcidoid, Pachy crepoideus dubius Ashmead* from both Drosophila melanogaster ani D. hydei. Kieffer ( 1913) has described three species of hymenoptera from Africa collected by Silvestri and stated by him to be parasitic on Drosophila, species not given. They are Trichopria (Planopria) rhopalica (Diapriidae), Ashmeadopria drosophilae (Diapriidae), and the insect which forms the subject matter of the present investigation, Eucoila drosophilae (Figitidae).

There are in addition a number of predacious enemies among wasps, spiders, flies and beetles.

The present account is concerned with parasitism of various species of Drosophila by Eucoila drosophilae Kieff. The wasps were found b y Dr. w. P. Spencer who exposed traps in an effort to collect Drosophila at Long Lake, Ohio, in Sept. 1934 . Drosophila larvae from the trap gave a large number of pupae from which wasps emerged in considerable proportions. Since that time stock s have been maintained in culture on Drosophila melanogaster.

Studies on 4S and 5S RNA of HeLa cells

SECTION I

Section I is concerned with a partial sequence analysis conducted on 5S RNA from HeLa cells. Analysis of the oligonucleotide pattern after pancreatic ribonuclease digestion of a highly-purified preparation of 5S RNA gave results which were in general agreement with those published for KB cells, both with respect to the identity and the frequency of the partial sequences. However, the presence of a trinucleotide not found in the KB 5S pattern, together with the reproducibly much lower than expected molar yield of the larger oligonucleotides strongly suggested the occurrence of alternate sequences at various sites in the 5S molecules of human cells. The presence of ppGp and pppGp at the 5'-terminus of HeLa 5S RNA was clearly demonstrated. The implications of this finding with regard to the origin of 5S RNA are discussed.

SECTION II

In Section II the proportion of the HeLa cell genome complementary to tRNA was investigated by using RNA- DNA hybridization. The value for saturation of the HeLa DNA by tRNA was found to be 1.1 x 10^-5, which corresponds to about 4900 sites for tRNA per HeLa cell in an exponentially growing culture. Analysis of the nucleotide composition of the hybridized tRNA revealed significant differences from the nucleotide composition of the input tRNA, with the purine to pyrimidine ratio indicating, however, that these differences were not produced by excessive RNase attack of the hybrid. The size of the hybridized tRNA was only moderately smaller than that of the input RNA; the average S value in formaldehyde was 2.7 (corresponding to a length of about 65 nucleotides), suggesting that a relatively small portion near the ends of the hybridized 4S chains had been removed by RNase.

SECTION III

The proportion of the HeLa cell genome complementary to 5S RNA was investigated by using RNA-DNA hybridization. The value for saturation of the HeLa DNA by 5S RNA was found to be 2.3 x 10^-5, which corresponds to about 7,000 sites for 5S RNA per HeLa cell in an exponentially growing culture. Analysis of the nucleotide composition of the hybridized 5S RNA revealed no significant difference from the nucleotide composition of the input RNA. At the RNA to DNA input ratio of 1:1000, the average S value in formaldehyde of the hybridized 5S RNA corresponded to a polynucleotide chain about two-thirds the size of the input RNA.

I. Studies on bacteriophage DNA's and minicircular DNA's in M. lysodeikticus and E. coli 15. II. Flow dichroism of DNA solutions

Part I

Chapter 1.....A physicochemical study of the DNA molecules from the three bacteriophages, N1, N5, and N6, which infect the bacterium, M. lysodeikticus, has been made. The molecular weights, as measured by both electron microscopy and sedimentation velocity, are 23 x 10⁶ for N5 DNA and 31 x 10⁶ for N1 and N6 DNA's. All three DNA's are capable of thermally reversible cyclization. N1 and N6 DNA's have identical or very similar base sequences as judged by membrane filter hybridization and by electron microscope heteroduplex studies. They have identical or similar cohesive ends. These results are in accord with the close biological relation between N1 and N6 phages. N5 DNA is not closely related to N1 or N6 DNA. The denaturation T_m of all three DNA's is the same and corresponds to a (GC) content of 70%. However, the buoyant densities in CsCl of Nl and N6 DNA's are lower than expected, corresponding to predicted GC contents of 64 and 67%. The buoyant densities in Cs₂SO₄ are also somewhat anomalous. The buoyant density anomalies are probably due to the presence of odd bases. However, direct base composition analysis of N1 DNA by anion exchange chromatography confirms a GC content of 70%, and, in the elution system used, no peaks due to odd bases are present.

Chapter 2.....A covalently closed circular DNA form has been observed as an intracellular form during both productive and abortive infection processes in M. lysodeikticus. This species has been isolated by the method of CsC1-ethidium bromide centrifugation and examined with an electron microscope.

Chapter 3.....A minute circular DNA has been discovered as a homogeneous population in M. lysodeikticus. Its length and molecular weight as determined by electron microscopy are 0.445 μ and 0.88 x 10⁶ daltons respectively. There is about one minicircle per bacterium.

Chapter 4.....Several strains of E. coli 15 harbor a prophage. Viral growth can be induced by exposing the host to mitomycin C or to uv irradiation. The coliphage 15 particles from E. coli 15 and E, coli 15 T^- appear as normal phage with head and tail structure; the particles from E. coli 15 TAU are tailless. The complete particles exert a colicinogenic activity on E.coli 15 and 15 T^-, the tailless particles do not. No host for a productive viral infection has been found and the phage may be defective. The properties of the DNA of the virus have been studied, mainly by electron microscopy. After induction but before lysis, a closed circular DNA with a contour length of about 11.9 μ is found in the bacterium; the mature phage DNA is a linear duplex and 7.5% longer than the intracellular circular form. This suggests the hypothesis that the mature phage DNA is terminally repetitious and circularly permuted. The hypothesis was confirmed by observing that denaturation and renaturation of the mature phage DNA produce circular duplexes with two single-stranded branches corresponding to the terminal repetition. The contour length of the mature phage DNA was measured relative to φX RFII DNA and λ DNA; the calculated molecular weight is 27 x 10⁶. The length of the single-stranded terminal repetition was compared to the length of φX 174 DNA under conditions where single-stranded DNA is seen in an extended form in electron micrographs. The length of the terminal repetition is found to be 7.4% of the length of the nonrepetitious part of the coliphage 15 DNA. The number of base pairs in the terminal repetition is variable in different molecules, with a fractional standard deviation of 0.18 of the average number in the terminal repetition. A new phenomenon termed "branch migration" has been discovered in renatured circular molecules; it results in forked branches, with two emerging single strands, at the position of the terminal repetition. The distribution of branch separations between the two terminal repetitions in the population of renatured circular molecules was studied. The observed distribution suggests that there is an excluded volume effect in the renaturation of a population of circularly permuted molecules such that strands with close beginning points preferentially renature with each other. This selective renaturation and the phenomenon of branch migration both affect the distribution of branch separations; the observed distribution does not contradict the hypothesis of a random distribution of beginning points around the chromosome.

Chapter 5....Some physicochemical studies on the minicircular DNA species in E. coli 15 (0.670 μ, 1.47 x 10⁶ daltons) have been made. Electron microscopic observations showed multimeric forms of the minicircle which amount to 5% of total DNA species and also showed presumably replicating forms of the minicircle. A renaturation kinetic study showed that the minicircle is a unique DNA species in its size and base sequence. A study on the minicircle replication has been made under condition in which host DNA synthesis is synchronized. Despite experimental uncertainties involved, it seems that the minicircle replication is random and the number of the minicircles increases continuously throughout a generation of the host, regardless of host DNA synchronization.

Part II

The flow dichroism of dilute DNA solutions (A₂₆₀≈0.1) has been studied in a Couette-type apparatus with the outer cylinder rotating and with the light path parallel to the cylinder axis. Shear gradients in the range of 5-160 sec.^-1 were studied. The DNA samples were whole, "half," and "quarter" molecules of T4 bacteriophage DNA, and linear and circular λb₂b₅c DNA. For the linear molecules, the fractional flow dichroism is a linear function of molecular weight. The dichroism for linear A DNA is about 1.8 that of the circular molecule. For a given DNA, the dichroism is an approximately linear function of shear gradient, but with a slight upward curvature at low values of G, and some trend toward saturation at larger values of G. The fractional dichroism increases as the supporting electrolyte concentration decreases.