Identification of larval sea basses (Centropristis spp.) using ribosomal DNA-specific molecular assays

The identification of sea bass (Centropristis) larvae to species is difficult because of similar morphological characters, spawning times, and overlapping species ranges. Black sea bass (Centropristis striata) is an important fishery species and is currently considered to be overfished south of Cape Hatteras, North Carolina. We describe methods for identifying three species of sea bass larvae using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays based on species-specific amplification of rDNA internal transcribed spacer regions. The assays were tested against DNA of ten other co-occurring reef fish species to ensure the assay's specificity. Centropristis larvae were collected on three cruises during cross-shelf transects and were used to validate the assays. Seventy-six Centropristis larva were assayed and 69 (91%) were identified successfully. DNA was not amplified from 5% of the larvae and identification was inconclusive for 3% of the larvae. Those assays can be used to identify sea bass eggs and larvae and will help to assess spawning locations, spawning times, and larval dispersal.

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)

Shoal manipulation as an effective solution to site specific beach management, examples from three South Carolina beaches

Soft engineering solutions are the current standard for addressing coastal erosion in the US. In South Carolina, beach nourishment from offshore sand deposits and navigation channels has mostly replaced construction of seawalls and groins, which were common occurrences in earlier decades. Soft engineering solutions typically provide a more natural product than hard solutions, and also eliminate negative impacts to adjacent areas which are often associated with hard solutions. A soft engineering solution which may be underutilized in certain areas is shoal manipulation. (PDF contains 4 pages)

Regulation of the fitness of Atlantic salmon (Salmo salar) by intra-specific competition amongst the juveniles

Factors affecting the fitness of juvenile salmon are discussed. Although fitness from the genetic point of view is defined as the relative capacity of carriers of a given genotype to transmit their genes to the gene pool of the following generations, growth and survival of individuals are also components of fitness, and are influenced by responses to competition, which is the major topic of this article including implications for management. In order to better understand the relationships of density-dependent survival in Newfoundland, egg depositions were manipulated experimentally in the Freshwater River. Figures demonstrate the relationship between stock (number of eggs per 100 m2 of river) and recruitment (number of smolts per l00 m2 of Atlantic salmon, and also the percentage survival from egg to smolt stage related to potential egg depositions.

Tests of some specific effect substances in duck nutrition. Translation from: Biologisace a chemisaze vyzivy svirat 5, 43-48, 1969.]

There is a constantly increasing collection of manufactured substances, whose effectiveness in the sustenance of ducks is under investigation. The author examines the effects of some substances already previously tested and also there were examined substances which had not hitherto been studied. The use of different supplements for late autumn fattening is studied through various experiments.

Detection of specific bacteria in water: implications of survival strategy

It is widely recognised that conventional culture techniques may underestimate true viable bacterial numbers by several orders of magnitude. The basis of this discrepancy is that a culture in or on media of high nutrient concentration is highly selective (either through ”nutrient shock” or failure to provide vital co-factors) and decreases apparent diversity; thus it is unrepresentative of the natural community. In addition, the non-culturable but viable state (NCBV) is a strategy adopted by some bacteria as a response to environmental stress. The basis for the non-culturable state is that cells placed in conditions present in the environment cannot be recultured but can be shown to maintain their viability. Consequently, these cells would not be detected by standard water quality techniques that are based on culture. In the case of pathogens, it may explain outbreaks of disease in populations that have not come into contact with the pathogen. However, the NCBV state is difficult to attribute, due to the failure to distinguish between NCBV and non-viable cells. This article will describe experiences with the fish pathogen Aeromonas salmonicida subsp. salmonicida and the application of molecular techniques for its detection and physiological analysis.

Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no convenient method of preparing viruses for study or for reagent production. One problem facing those attempting to use PCR-based methods for the detection of HuCVs and SRSVs is the lack of a suitable positive control substrate. This is particularly important when screening complex samples in which the levels of inhibitors present may significantly interfere with amplificiation. Regions within the RNA polymerase regions of two genetically distinct human caliciviruses have been amplified and used to produce recombinant baculoviruses which express RNA corresponding to the calicivirus polymerase. This RNA is being investigated as a positive control substrate for PCR testing, using current diagnostic primer sets. Recombinant baculovirus technology will enable efficient and cost-effective production of large quantities of positive control RNA with a specific known genotype. We consider the development of these systems as essential for successful screening and monitoring applications.

Stage-specific vertical distribution of Alaska plaice (Pleuronectes quadrituberculatus) eggs in the eastern Bering Sea

The stage-specific distribution of Alaska plaice (Pleuronectes quadrituberculatus) eggs in the southeastern Bering Sea was examined with collections made in mid-May in 2002, 2003, 2005, and 2006. Eggs in the early stages of development were found primarily offshore of the 40-m isobath. Eggs in the middle and late stages of development were found inshore and offshore of the 40-m isobath. There was some evidence that early-stage eggs occur deeper in the water column than late-stage eggs, although year-to-year variability in that trend was observed. Most eggs were in the later stages of development; therefore the majority of spawning is estimated to have occurred a few weeks before collection—probably April—and may be highly synchronized among local spawning areas. Results indicate that sampling with continuous underway fish egg collectors(CUFES) should be supplemented with sampling of the entire water column to ensure adequate samples of all egg stages of Alaska plaice. Data presented offer new information on the stage-dependent horizontal and vertical distribution of Alaska plaice eggs in the Bering Sea and provide further evidence that the early life history stages of this species are vulnerable to near-surface variations in hydrographical conditions and climate forcing.

Analysis of sex-specific spawning biomass per recruit of the sailfish (Istiophorus platypterus) in the waters off eastern Taiwan

Analyses of sex-specific yield per recruit and spawning stock biomass per recruit were conducted to evaluate the current status of the sailfish (Istiophorus platypterus) fishery in the waters off eastern Taiwan. Natural mortality rates estimated from Pauly’s empirical equation were 0.26/yr for females and 0.27/yr for males. The current fishing mortality rates were estimated as 0.24/yr and 0.43/yr for females and males, respectively, which are much lower than the estimated F0 .1 (0.62/yr and 0.79/yr for females and males, respectively) and FSSB40 (0.46/yr for females) which are commonly used as target reference points in fisheries management. The effects of the fishing mortality, natural mortality, and age at first capture on the estimates of biological reference points were evaluated by using the Monte Carlo simulation. The results indicate that failure to consider the uncertainty in parameters such as natural mortality or age at first capture may lead to the improper estimation of biological reference points. This study indicates the possibility of current fishing mortality exceeding the target biological reference points may be negligible for sailfish in the waters off eastern Taiwan. However, in view of the recent rapid increase in fishing effort, it is evident that the stock status and development of the fishery need to be closely monitore

Age-specific movement patterns of sablefish (Anoplopoma fimbria) in Alaska

Over 34,000 age 0–2 juvenile sablefish (Anoplopoma fimbria) were tagged and released in southeast Alaska waters during 1985–2005. The data set resulting from this tagging study was unusual because of its time span (20 years) and because age could be reliably inferred from release length (i.e., tagged and released fish were of known age); thus, age-specific movement patterns could be examined. The depth- and area-related recovery patterns supported the concepts that sablefish move to deeper water with age and migrate counterclockwise in the Gulf of Alaska. Availability to the fishery increased rapidly for fish of younger ages, peaked at age 5 to 6, and then gradually declined as sablefish moved deeper with age. Decreased availability with age may occur because of lower fishing effort in deep water and could have substantial implications for sablef ish stock assessments because “domeshaped” availability influences the reliability of abundance estimates. The area-related recovery pattern was not affected by year-class strength; i.e., there was no significant densitydependent relationship.

Species-specific effects of four preservative treatments on oocytes and ovarian material of Atlantic cod (Gadus morhua), haddock (Melanogrammus aeglefinus), and American plaice (Hippoglossoides platessoides)

The lack of information concerning the preservation of ovarian material of fish species inhibits standardization of methods for determining fecundity and measuring oocytes. The effects of four preservatives (10% phosphate-buffered formalin, modified Gilson’s solution, 70% ethanol, and freezing) on ovarian material weight and oocyte size were quantified for prespawning Atlantic cod (Gadus morhua), haddock (Melanogrammus aeglefinus), and American plaice (Hippoglossoides platessoides). Effects of preservation were similar between Atlantic cod and haddock but different between Atlantic cod and American plaice for nearly all comparisons. Although all treatments affected the weight of ovarian material, freezing caused the most change and formalin caused the least. Such signif icant species-specific effects should be quantified in the calculation of life history characteristics, such as fecundity, to minimize error. This is one of few studies dedicated to evaluating the effects of preservation on oocytes and ovarian material and is the first to evaluate multiple preservatives on species.

Is epizootic ulcerative syndrome (EUS) specific fungus of fishes a primary pathogen?: an opinion

Earlier findings on epizootic ulcerative syndrome (EUS) and the present observation of the authors on transmission of EUS to snakehead (Channa sp.) without skin damage provide evidence to suggest that the invasive fungus associated with EUS is a primary pathogen.

Influence of nutrient source on specific dynamic action of Pearl Spot, Etroplus suratensis (Bloch)

The magnitude of apparent specific dynamic action (SDA), the maximum rate of oxygen consumption and the length of time that the rate of oxygen uptake remained elevated above the prefeeding level were measured in the Pearl Spot, Etroplus suratensis, fed isonitrous test diets (D 1 - D 4 ) with varying nutrient sources. Irrespective of the diets, the metabolic rate increased immediately after feeding and reached the maximum within 3 to 4 hours. The source of nutrients in the diet significantly altered the magnitude of SDA. It was maximum (91.76% and 129.56%) for those fed on diets D 2 and D 3 and minimum 46.47% and 50.30% for those fed on diets D 1 and D 4 , respectively.