Report on Marine Mammal Unusual Mortality Event UMESE0501Sp: Multispecies Mass Stranding of Pilot Whales (Globicephala macrorhynchus), Minke Whale (Balaenoptera acutorostrata), and Dwarf Sperm Whales (Kogia sima) in North Carolina on 15-16 January 2005

On 15-16 January 2005, three offshore species of cetaceans (33 short-finned pilot whales, Globicephala macrorhynchus, one minke whale, Balaenoptera acutorostrata, and two dwarf sperm whales, Kogia sima) stranded alive on the beaches of North Carolina. The pilot whales stranded near Oregon Inlet, the minke whale in northern North Carolina, and the dwarf sperm whales near Cape Hatteras. Live strandings of three species in one weekend was unique in North Carolina and qualified as an Unusual Mortality Event. Gross necropsies were conducted on 16-17 January 2005 on 27 pilot whales, two dwarf sperm whales, and the minke whale. Samples were collected for clinical pathology, parasitology, gross pathology, histopathology, microbiology and serology. There was variation in the number of animals sampled for each collection type, however, due to carcasses washing off the beach or degradation in carcass condition during the course of the response. Comprehensive histologic examination was conducted on 16 pilot whales, both dwarf sperm whales, and the minke whale. Limited organ or only head tissue suites were obtained from nine pilot whales. Histologic examination of tissues began in February 2005 and concluded in December 2005 when final sampling was concluded. Neither the pilot whales nor dwarf sperm whales were emaciated although none had recently ingested prey in their stomachs. The minke whale was emaciated; it was likely a dependent calf that became separated from the female. Most serum biochemistry abnormalities appear to have resulted from the stranding and indicated deteriorating condition from being on land for an extended period. Three pilot whales had clinical evidence of pre-existing systemic inflammation, which was supported by histopathologic findings. Although gross and histologic lesions involving all organ systems were noted, consistent lesions were not observed across species. Verminous pterygoid sinusitis and healed fishery interactions were seen in pilot whales but neither of these changes were causes of debilitation or death. In three pilot whales and one dwarf sperm whale there was evidence of clinically significant disease in postcranial tissues which led to chronic debilitation. Cardiovascular disease was present in one pilot whale and one dwarf sperm whale; musculoskeletal disease and intra-abdominal granulomas were present in two pilot whales. These lesions were possible, but not definitive, causal factors in the stranding. Remaining lesions were incidental or post-stranding. The minke whale and three of five tested pilot whales had positive morbillivirus titers (≥1:8 with one at >1:256), but there was no histologic evidence of active viral infection. Parasites (nematodes, cestodes, and trematodes) were collected from 26 pilot whales and two dwarf sperm whales. Sites of collection included stomach, nasal/pterygoid, peribullar sinuses, blubber, and abdominal cavity. Parasite species, locations and loads were within normal limits for free-ranging cetaceans and were not considered causative for the stranding event. Gas emboli lesions which were considered consistent with or diagnostic of sonarassociated strandings of beaked whales or small cetaceans were not found in the whales stranded as part of UMESE0501Sp. Twenty-five heads were examined with nine specific anatomic locations of interest: extramandibular fat, intramandibular fat, auditory meatus, peribullar acoustic fat, peribullar soft tissue, peribullar sinus, pterygoid sinus, melon, and brain. The common finding in all examined heads was verminous pterygoid sinusitis. Intramandibular adipose tissue reddening, typically adjacent to the vascular plexus, was observed in some individuals and could represent localized hemorrhage resulting from vascular rete rupture, hypostatic congestion, or erythrocyte rupture during the freeze/thaw cycle. One cetacean had peracute to acute subdural hemorrhage that likely occurred from thrashing on the beach post-stranding, although its occurrence prior to stranding cannot be excluded. Information provided to NMFS by the U.S. Navy indicated routine tactical mid-frequency sonar operations from individual surface vessels over relatively short durations and small spatial scales within the area and time period investigated. No marine mammals were detected by marine mammal observers on operational vessels; standard operating procedure for surface naval vessels operating mid-frequency sonar is the use of trained visual lookouts using high-powered binoculars. Sound propagation modeling using information provided to NMFS indicated that acoustic conditions in the vicinity likely depended heavily on position of the receivers (e.g., range, bearing, depth) relative to that of the sources. Absent explicit information on the location of animals meant that it was not possible to estimate received acoustic exposures from active sonar transmissions. Nonetheless, the event was associated in time and space with naval activity using mid-frequency active sonar. It also had a number of features in common (e.g., the “atypical” distribution of strandings involving multiple offshore species, all stranding alive, and without evidence of common infectious or other disease process) with other sonar-related cetacean mass stranding events. Given that this event was the only stranding of offshore species to occur within a 2-3 day period in the region on record (i.e., a very rare event), and given the occurrence of the event simultaneously in time and space with a naval exercise using active sonar, the association between the naval sonar activity and the location and timing of the event could be a causal rather than a coincidental relationship. However, evidence supporting a definitive association is lacking, and, in particular, there are differences in operational/environmental characteristics between this event and previous events where sonar has apparently played a role in marine mammal strandings. This does not preclude behavorial avoidance of noise exposure. No harmful algal blooms were present along the Atlantic coast south of the Chesapeake Bay during the months prior to the event. Environmental conditions, including strong winds, changes in upwelling- to downwelling-favorable conditions, and gently sloping bathymetry, were consistent with conditions which have been correlated with other mass strandings. In summary, we did not find commonality in gross and histologic lesions that would indicate a single cause for this stranding event. Three pilot whales and one dwarf sperm whale had debilitating conditions identified that could have contributed to stranding, one pilot whale had a debilitating condition (subdural hemorrhage) that could have been present prior to or resulting from stranding. While the pilot and dwarf sperm whale strandings may have had a common cause, the minke whale stranding was probably just coincidental. On the basis of examination of physical evidence in the affected whales, however, we cannot definitively conclude that there was or was not a causal link between anthropogenic sonar activity or environmental conditions (or a combination of these factors) and the strandings. Overall, the cause of UMESE0501Sp in North Carolina is not and likely will not be definitively known. (PDF contains 240 pages)

Marine Flora and Fauna of the eastern United States: Acanthocephala

The phylum Acanthocephala (intestinal worm parasites of vertebrates) of the Atlantic coast of the United States comprises 43 species and 20 genera belonging to three orders: Echinorhynchida, Neoechinorhynchida, and Polymorphida. Adults are exclusively intestinal parasites of vertebrates. This study includes those species found in vertebrates of marine and estuarine environments along the North American Atlantic coast between Maine and Texas. Species that can be found within that geographical range and those that typically infect freshwater fishes but that are occasionally present in marine or estuarine hosts are also included. The taxonamy, anatomy, natural history, and ecology of the phylum Acanthocephala are discussed, and an illustrated key to the genera is presented. Techniques, an annotated systematic treatment of all 43 species, and a systematic index are included. No systematic decisions will be made at this time, but areas where such decisions are pending will be indicated and discussed for future reports. (PDF file contains 32 pages.)

Marine Flora and Fauna of the Eastern United States: Dicyemida

This manual treats the six species of dicyemid mesozoans that have been reported in three species of hosts (Octopus vulgaris, O. joubini, and O. briareus) from the eastern coast of North America and the Gulf of Mexico, including the Florida Keys. All are parasites of species of Octopus and are in the genus Dicyema, family Dicyemidae. In the introduction, the life cycle, as known, and the general morphology of dicyemids are briefly described, and methods are given for collecting and preparing material for study. These are followed by a key to species and by an annotated checklist, which includes data, some hitherto unpublished, on their known prevalence in hosts from various localities including Bimini and Bermuda.(PDF file contains 20 pages.)

Prevalence, intensity, longevity, and persistence of Anisakis sp. larvae and Lacistorhynchus tenuis metacestodes in San Francisco striped bass

Thirteen hundred and seventy-three striped bass, Marone saxatilis, were collected from the San Francisco Bay-Delta area to correlate host diet with parasitic infections and to determine the prevalence, intensity, longevity, and persistence of larval Anisakis sp. nematodes and the metacestode Lacistorhynchus tenuis. There is an increase in the prevalence and intensity of Anisakis sp. and in the intensity of L. tenuis with increase of age of the host. These increases are probably related to the diet and the persistence of tbe parasites. The infections of both species are overdispersed. San Francisco Bay striped bass are an incompatible host for both species of parasites. Degenerated Anisakis sp. will remain in lhe host for at least 8 months and L. tenuis metacestodes for 22 months. The occurrence of several other species of parasites and a tumor are also reported. (PDF file contains 10 pages.)

The feeding ecology of some zooplankters that are important prey items of larval fish

Diets of 76 species of fish larvae from most oceans of the world were inventoried on the basis of information in 40 published studies. Although certaln geographlc, size- and taxon-specific patterns were apparent, certain zooplankton taxa appeared in the diets of larvae of a variety of fish species in numerous localities. Included were six genera of calanoid copepods (Acartia, Calanus, Centropages, Paracalanus, Pseudocaianus, Temora), three genera of cyclopoid copepods (Corycaeus, Oilhona, Oncata), harpacticoid copepods, copepod nauplii, tintinoids, cladocerans of the genera Evadne and Podon, barnacle nauplii, gastropod larvae, pteropods of the genus Limacina, and appendicularians. Literature on feeding habits of these zooplankters reveals that most of the copepods are omnivorous, feeding upon both phytoplankton and other zooplankton. Some taxa, such as Calanus, Paracalanus, Pseudocalanus, and copepod nauplii appear to be primarily herbivorous, while others, such as Acartia, Centropages, Temora, and cyclopoids exhibit broad omnivory or carnivory. The noncopepod zooplankters are primarily filter-feeders upon pbytoplankton and/or bacterioplankton. Despite the importance of zooplankters in larval fish food webs, spectic knowledge of the feeding ecology of many taxa is poor. Further, much present knowledge comes only from laboratory investigations that may not accurately portray feeding habits of zooplankters in nature. Lack of knowledge of the feeding ecology of many abundant zooplankters, which are also important in larval fish food webs, precludes realistic understanding of pelagic ecosystem dynamics. (PDF file contains 34 pages.)

Inoculation of Triatoma Virus (Dicistroviridae: Cripavirus) elicits a non-infective immune response in mice

Background: Dicistroviridae is a new family of small, non-enveloped, +ssRNA viruses pathogenic to both beneficial arthropods and insect pests. Little is known about the dicistrovirus replication mechanism or gene function, and any knowledge on these subjects comes mainly from comparisons with mammalian viruses from the Picornaviridae family. Due to its peculiar genome organization and characteristics of the per os viral transmission route, dicistroviruses make good candidates for use as biopesticides. Triatoma virus (TrV) is a pathogen of Triatoma infestans (Hemiptera: Reduviidae), one of the main vectors of the human trypanosomiasis disease called Chagas disease. TrV was postulated as a potential control agent against Chagas' vectors. Although there is no evidence that TrV nor other dicistroviruses replicate in species outside the Insecta class, the innocuousness of these viruses in humans and animals needs to be ascertained. Methods: In this study, RT-PCR and ELISA were used to detect the infectivity of this virus in Mus musculus BALB/c mice. Results: In this study we have observed that there is no significant difference in the ratio IgG2a/IgG1 in sera from animals inoculated with TrV when compared with non-inoculated animals or mice inoculated only with non-infective TrV protein capsids. Conclusions: We conclude that, under our experimental conditions, TrV is unable to replicate inmice. This study constitutes the first test to evaluate the infectivity of a dicistrovirus in a vertebrate animal model.

Recovery of the stocks of the European eel (Anguilla anguilla) by spawner enhancement

The production of healthy high quality female European eel in recycle systems is proposed as a means to secure sufficient numbers of silver eel for spawning migration in order to meet the requirements of the European Commission’s proposal for a Regulation for the recovery of the stock of the European eel. Main advantages besides checks for parasites and viral diseases and avoidance of elevated levels of specific pollutants are the easily controllable numbers of spawners to be released and a reduction of labour and costs that will occur when acting along the lines of the Commission’s proposal.

Ecology and fishery in the Cross River Estuary - a research project

A literature review of 50 titles including nearly all relevant publications ensures adequate basis on the present level of knowledge. The proposal includes (a) the determination of the biozoenosis and selected environmental factors, and (b) of fishery and stock data of the main fish and shellfish species. The ecological research studies physical and chemical variables of the estuarine waters (flow velocity and direction, water temperature, conductivity, pH, dissolved oxygen, salinity, nutrients such as ammonium, nitrite, phosphate, silicate, pollutants such as hybrocarbons, pesticides and heavy metals, biochemical oxygen demand, chemical oxygen demand), plankton (bacterio-phyto-and zooplankton), benthos, sediment. The fishery biological and fishery investigations include: number of villages and fishermen, number of boats and gears by type, length and weight data of the main fishery objects with concentration on the shrimps, species and numbers of fish parasites. The ecological variables were monitored at fixed stations on sections in the Cross-River Estuary, Calabar and Great Kwara Rivers two times per month during spring and neap tides. The fishery biological and fishery variables were obtained during spring and neap tide too. For the determination of the detailed methodology the ecological and fishery part of the progeamme should be started with frame surveys based on a larger number of stations. These frame surveys should be repeated from time to time. Both parts of the programme are based on three years duration. It seems already appropriate to continue the work with selected representative stations, villages and variables in form of a long-term data chain

A review of diseases associated with cage culture systems: diagnosis and control in smale scale fish farming

Fish cage culture is a rapid aquacultural practice of producing fish with more yield compared to traditional pond culture. Several species cultured by this method include Cyprinus carpio, Orechromis niloticus, Sarotherodon galilaeus, Tilapia zilli, Clarias lazera, C. gariepinus, Heterobranchus bidorsalis, Citharinus citharus, Distochodus rostratus and Alestes dentes. However, the culture of fish in cages has some problems that are due to mechanical defects of the cage or diseases due to infection. The mechanical problems which may lead to clogged net, toxicity and easy access by predators depend on defects associated with various types of nets which include fold sieve cloth net, wire net, polypropylene net, nylon, galvanized and welded net. The diseases problems are of two types namely introduced diseases due to parasites. The introduced parasites include Crustaseans, Ergasilus sp. Argulus africana, and Lamprolegna sp, Helminth, Diplostomulum tregnna: Protozoan, Trichodina sp, Myxosoma sp, Myxobolus sp. the second disease problems are inherent diseases aggravated by the very rich nutrient environment in cages for rapid bacterial, saprophytic fungi, and phytoplanktonic bloom resulting in clogging of net, stagnation of water and low biological oxygen demand (BOD). The consequence is fish kill, prevalence of gill rot and dropsy conditions. Recommendations on routine cage hygiene, diagnosis and control procedures to reduce fish mortality are highlighted

Some potential occupational and environmental hazards associated with fish pond production in Nigeria

The paper highlights the concept of information and the significance of environmental and occupational hazards associated with pond fish production in Nigeria and discuss the possible options for the ways forward. The major raw material used in fish production system is the organic manure (cow dung, poultry droppings, porcine manure etc) that serves as substrate for heterotrophic production of bacteria and protozoa, which act as food for zooplankton and the fish. The pathogenic organisms (viruses, bacteria, protozoa's, and parasites), are noted for the potential hazard to the fish handlers and consumers. Nine species from seven genera of bacteria associated with fish diseases are found to have association with diseases of human such as typhoid fever, bacillary dysentery and other gastrointestinal tract related problems. Also the environmental contaminants in pond fish production become important because of its significance to consumers' acceptance of the fish products

Internationaler BMB/ICES-Workshop "Fischkankheiten und Parasiten in der Ostsee"

From 25 November until 8 Deeember 1994, a sea-going workshop with the above title was carried out on board RV "Walther Herwig IlI" under the umbrella of the Baltie Marine Biologists (BMlJ) and the International Council for the Exploration of the Sea (ICES). Under the co-convenorship of the first two authors of this paper, 11 seientists from 8 of the 9 countries bordering the Baltic Sea (except Sweden) actively involved in research monitoring into fish diseases/parasites participated. The main reason for holding the workshop was that, although fish disease monitoring programmes on the abundance and spatial distribution of fish diseases/parasites are being carried out by some of the countries bordering the Baltic Sea, there is still a striking lack of seientific information as compared to other seas, such as the North Sea. Moreover, since new fish disease monitoring programmes have recently been started (mainly by the new Baltic republics Latvia and Estonia, which recently became ICES-member countries) or are planned for the near future (Lithuania), BMB as weil as ICES felt the need for a interealibration and standardization of methodologies applied during fish disease/parasite surveys for the specific conditions in the Baltic Sea. Therefore, the major objectives of the workshop were -to obtain information about the disease/parasite prevalence in Baltic (Platichthys flesus) and other abundant fish species along a transect from the western (Mecklenhurg Bight) to the eastern (Gulf of Finland) Baltic Sea to be used as a baseline for future studies, -to intercalibrate methodologies applied for sampling, diagnosis of diseases/parasites, reporting and analyses of disease data, -if necessary, to recommend standard methodologies to be used for fish disease/parasite monitoring programmes under the specific conditions in the Baltic Sea. Although the data have yet not been completely analyzed, there is indication for some pronounced spatial trends regarding the prevalence of certain diseases/parasites, mainly for Iymphocystis and skin ulcerations in flounder but also for diseases and parasites of cod (Gadus morhua) which can be seen in figures 2 and 4. Recommendations for standard methodologies will later be published elsewhere.

Bright-field and fluorescence chip-scale microscopy for biological imaging

Optical microscopy is an essential tool in biological science and one of the gold standards for medical examinations. Miniaturization of microscopes can be a crucial stepping stone towards realizing compact, cost-effective and portable platforms for biomedical research and healthcare. This thesis reports on implementations of bright-field and fluorescence chip-scale microscopes for a variety of biological imaging applications. The term “chip-scale microscopy” refers to lensless imaging techniques realized in the form of mass-producible semiconductor devices, which transforms the fundamental design of optical microscopes.

Our strategy for chip-scale microscopy involves utilization of low-cost Complementary metal Oxide Semiconductor (CMOS) image sensors, computational image processing and micro-fabricated structural components. First, the sub-pixel resolving optofluidic microscope (SROFM), will be presented, which combines microfluidics and pixel super-resolution image reconstruction to perform high-throughput imaging of fluidic samples, such as blood cells. We discuss design parameters and construction of the device, as well as the resulting images and the resolution of the device, which was 0.66 µm at the highest acuity. The potential applications of SROFM for clinical diagnosis of malaria in the resource-limited settings is discussed.

Next, the implementations of ePetri, a self-imaging Petri dish platform with microscopy resolution, are presented. Here, we simply place the sample of interest on the surface of the image sensor and capture the direct shadow images under the illumination. By taking advantage of the inherent motion of the microorganisms, we achieve high resolution (~1 µm) imaging and long term culture of motile microorganisms over ultra large field-of-view (5.7 mm × 4.4 mm) in a specialized ePetri platform. We apply the pixel super-resolution reconstruction to a set of low-resolution shadow images of the microorganisms as they move across the sensing area of an image sensor chip and render an improved resolution image. We perform longitudinal study of Euglena gracilis cultured in an ePetri platform and image based analysis on the motion and morphology of the cells. The ePetri device for imaging non-motile cells are also demonstrated, by using the sweeping illumination of a light emitting diode (LED) matrix for pixel super-resolution reconstruction of sub-pixel shifted shadow images. Using this prototype device, we demonstrate the detection of waterborne parasites for the effective diagnosis of enteric parasite infection in resource-limited settings.

Then, we demonstrate the adaptation of a smartphone’s camera to function as a compact lensless microscope, which uses ambient illumination as its light source and does not require the incorporation of a dedicated light source. The method is also based on the image reconstruction with sweeping illumination technique, where the sequence of images are captured while the user is manually tilting the device around any ambient light source, such as the sun or a lamp. Image acquisition and reconstruction is performed on the device using a custom-built android application, constructing a stand-alone imaging device for field applications. We discuss the construction of the device using a commercial smartphone and demonstrate the imaging capabilities of our system.

Finally, we report on the implementation of fluorescence chip-scale microscope, based on a silo-filter structure fabricated on the pixel array of a CMOS image sensor. The extruded pixel design with metal walls between neighboring pixels successfully guides fluorescence emission through the thick absorptive filter to the photodiode layer of a pixel. Our silo-filter CMOS image sensor prototype achieves 13-µm resolution for fluorescence imaging over a wide field-of-view (4.8 mm × 4.4 mm). Here, we demonstrate bright-field and fluorescence longitudinal imaging of living cells in a compact, low-cost configuration.

The possibilities and dangers in using malachite green in pisciculture.[Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod No.3 17-21, 1969.]

For many years a dye has been used in the practise of pisciculture and pond management, which is known by the name of malachite green, and is used to combat fungus on fish, fish eggs and external animal parasites on fish. The authors describe the problem of the application of malachite green in pisciculture and undertake special, complementary research.

Keys to the larvae of the species of the genera Diamesa, Eukiefferiella, Orthocladius, Cricotopus, Psectrocladius and Chaetocladius. [Translation from: Larvae and pupae of midges of the subfamily Orthocladiinae of the fauna of the USSR (Diptera, Chironomidae = Tendipedidae) (ed. V.Y. Pankratova) p76-8,151-152,173-174,188-189. Leningrad, Nauka, 1970. ]

Eudiaptomus vulgaris Schmeil is the most abundant copepod in Lake Maggiore and forms also, in respect to other entomostraca, the most important element, through its average biomass and because it is fairly numerous throughout the year. Plankton samples collected in a systematic and quantitative way, gave the opportunity to study some aspects of the dynamics of the population of this copepod, in safety in view of the uncertainty which in this kind of study can ensue when samples are taken only at a single station - in consequence of the changes in size of population between different water masses. The results of the biometrical observations are of the population of Eudiaptomus vulgaris is presented.

Ecological significance of seasonal variation in clutch-size of Arctodiaptomus bacillifer in alpine lakes of high altitudes (Val Bognanco) [Translation from: Bollettino di Zoologia 37 491-492, 1970]

This brief paper describes the significance of seasonal variation in clutch-size of the copepod Arctodiaptomus bacillifer in alpine lakes of high altitudes (Val Bognanco). Seasonal dynamics of the zooplankton of these lakes was studied during summer and autumn of 1968 and 1969and results are summarised.