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)

Development of kelp rockfish, Sebastes atrovirens (Jordan and Gilbert 1880), and brown rockfish, S. auriculatus (Girard 1854), from birth to pelagic juvenile stage, with notes on early larval development of black-and-yellow rockfish, S. chrysomelas (Jordan and Gilbert 1880), reared in the laboratory (Pisces: Sebastidae).

Larval kelp (Sebastes atrovirens), brown (S. auriculatus), and blackand-yellow (S. chrysomelas) rockfish were reared from known adults, to preflexion stage, nine days after birth for S. chrysomelas, to late postflexion stage for S. atrovirens, and to pelagic juvenile stage for S. auriculatus. Larval S. atrovirens and S. chrysomelas were about 4.6 mm body length (BL) and S. auriculatus about 5.2 mm BL at birth. Both S. atrovirens and S. auriculatus underwent notochord flexion at about 6–9 mm BL. Sebastes atrovirens transform to the pelagic juvenile stage at about 14–16 mm BL and S. auriculatus transformed at ca. 25 mm BL. Early larvae of all three species were characterized by melanistic pigment dorsally on the head, on the gut, on most of the ventral margin of the tail, and in a long series on the dorsal margin of the tail. Larval S. atrovirens and S. auriculatus developed a posterior bar on the tail during the flexion or postflexion stage. In S. atrovirens xanthic pigment resembled the melanistic pattern throughout larval development. Larval S. auriculatus lacked xanthophores except on the head until late preflexion stage, when a pattern much like the melanophore pattern gradually developed. Larval S. chrysomelas had extensive xanthic pigmentation dorsally, but none ventrally, in preflexion stage. All members of the Sebastes subgenus Pteropodus (S. atrovirens, S. auriculatus, S. carnatus, S. caurinus, S. chrysomelas, S. dalli, S. maliger, S. nebulosus, S. rastrelliger) are morphologically similar and all share the basic melanistic pigment pattern described here. Although the three species reared in this study can be distinguished on the basis of xanthic pigmentation, it seems unlikely that it will be possible to reliably identify field-collected larvae to species using traditional morphological and melanistic pigmentation characters. (PDF file contains 36 pages.)

White spot syndrome virus (WSSV) transmission risk through infected cooked shrimp products assessed by polymerase chain reaction (PCR) and bio-inoculation studies

The aim of the study was to evaluate the resistance of white spot syndrome virus (WSSV) in shrimps (Penaeus monodon) to the process of cooking. The cooking was carried out at 1000C six different durations 5, 10, 15, 20, 25 and 30 min. The presence of WSSV was tested by single step and nested polymerase chain reaction (PCR). In the single step PCR, the primers 1s5 & 1a16 and IK1 & IK2 were used. While in the nested PCR, primers IK1 &IK2 – IK3 & IK4 were used for the detection of WSSV. WSSV was detected in the single step PCR with the primers 1s5 and 1a16 and the nested PCR with the primers IK1 and IK2 – IK3 & IK4 from the cooked shrimp samples. The cooked shrimps, which gave positive results for WSSV by PCR, were further confirmed for the viability of WSSV by conducting the bio-inoculation studies. Mortality (100%) was observed within 123 h of intra-muscular post injection (P.I) into the live healthy WSSV-free shrimps (P. monodon). These results show that the WSSV survive the cooking process and even infected cooked shrimp products may pose a transmission risk for WSSV to the native shrimp farming systems.

Yellow (Perca flavescens) and Eurasian (P. fluviatilis) perch distinguished in fried fish samples by DNA analysis

DNA techniques are increasingly used as diagnostic tools in many fields and venues. In particular, a relatively new application is its use as a check for proper advertisement in markets and on restaurant menus. The identification of fish from markets and restaurants is a growing problem because economic practices often render it cost-effective to substitute one species for another. DNA sequences that are diagnostic for many commercially important fishes are now documented on public databases, such as the National Center for Biotechnology Information’s (NCBI) GenBank.1 It is now possible for most genetics laboratories to identify the species from which a tissue sample was taken without sequencing all the possible taxa it might represent.

Feeding habits of the dwarf weakfish (Cynoscion nannus) off the coasts of Jalisco and Colima, Mexico

Sciaenids from the Pacific coast of Mexico are used as a second-class fish species for human consumption (Aguilar-Palomino et al., 1996). The dwarf weakfish (Cynoscion nannus) (Castro-Aguirre and Arvizu-Martínez, 1976) is often caught as bycatch in the shrimp fishery but, because of its small size (<27 cm TL, total length), it is not considered a valuable resource. This species can be found in great numbers in waters between 100 and 812 m (Allen and Robertson, 1994; Fischer et al., 1995) associated with the soft-bottom regions off the coast of Jalisco and Colima (González-Sansón et al., 1997).

Preservation of fish by freezing and glazing. Pt. 2. Keeping quality of fish with particular reference to yellow discolouration and other allied organoleptic changes on prolonged storage

Organoleptic observations of quick, slow and block frozen, glazed and stored fish were recorded at regular intervals. Glazing was renewed at intervals of four weeks. Development of yellow discolouration in the case of white pomfret was followed. Keeping quality of glazed fish was better than unglazed frozen fish. Yellow discolouration could be controlled by ascorbic acid for 42 months and by a mixture of sodium chloride and glucose for 52 months.

Growth performance and nutrient utilization of catfish Clarias gariepinus fed varying inclusion level of fermented unsieved yellow maize

This study was conducted using 150 fish of Clarias gariepinus to investigate the growth performance and nutrient utilization of Clarias gariepinus fed five treatment diets containing varying inclusion level of fermented unsieved maize. The diets were grouped into CT, T1, T2, T3, and T4 with inclusion levels of 25%, 50%, 75%, and 100% of fermented unsieved maize respectively. Highest weight gain was recorded in T4 with value of 10.24 and lowest weight was recorded in CT with 9.17. High FCR were observed in T2 with value of 0.70 and lower value was observed in T4 with value of 0.62. While, T2, T3, and T4 have highest survival rates with values of 90% in each treatment CT and T1 recorded 80% and 70% respectively. There was a significant (p< 0.05) difference between the food conversion ratios treatment T4 with the best value and other treatments. There was a significant (p< 0.05) difference between the levels of fermented unsieved maize inclusion and the specific growth rate of the experimental fish. The highest value of protein level and feed efficiency were observed in T4 at significant difference level (p< 0.05) than other treatments. It was concluded that fermentation of maize in fish feed has positive effects on the nutritional value of the feed. It is recommend that fermented maize can replace raw maize in fish feed diet for growth performance. KEYWORDS: Fermentation, yellow maize, Clarias gariepinus, Fish, Feed.

Growth performance and nutrient utilization of catfish Clarias gariepinus fed varying inclusion level of fermented unsieved yellow maize

This study was conducted using 150 fish of Clarias gariepinus to investigate the growth performance and nutrient utilization of Clarias gariepinus fed five treatment diets containing varying inclusion level of fermented unsieved maize. The diets were grouped into CT, T1, T2, T3, and T4 with inclusion levels of 25%, 50%, 75%, and 100% of fermented unsieved maize respectively. Highest weight gain was recorded in T4 with value of 10.24 and lowest weight was recorded in CT with 9.17. High FCR were observed in T2 with value of 0.70 and lower value was observed in T4 with value of 0.62. While, T2, T3, and T4 have highest survival rates with values of 90% in each treatment CT and T1 recorded 80% and 70% respectively. There was a significant (p< 0.05) difference between the food conversion ratios treatment T4 with the best value and other treatments. There was a significant (p< 0.05) difference between the levels of fermented unsieved maize inclusion and the specific growth rate of the experimental fish. The highest value of protein level and feed efficiency were observed in T4 at significant difference level (p< 0.05) than other treatments. It was concluded that fermentation of maize in fish feed has positive effects on the nutritional value of the feed. It is recommend that fermented maize can replace raw maize in fish feed diet for growth performance.

Rapid detection of white spot syndrome virus (WSSV) of Penaeus monodon by latex agglutination test using monoclonal antibodies

Latex beads were sensitized with monoclonal antibodies (MAb) rose against VP28 of WSSV. The optimum concentration of MAb required to sensitize the latex beads was 125 µg/ml. The sensitized latex beads were used to detect WSSV from PCR-positive stomach tissue homogenates obtained from infected shrimp. Stomach tissue homogenates from WSSV-infected shrimp agglutinated the sensitized latex beads within 10 minutes, while uninfected samples did not produce any agglutination, although non-specific agglutinations were observed in some samples. The analytical sensitivity, analytical specificity, diagnostic sensitivity and diagnostic specificity of the (LAT) agglutination test were assessed. The analytical sensitivity of the test was 40 ng of purified WSSV (2 µg/ml). The sensitized latex beads did not agglutinate with normal shrimp tissue or MBV-infected tissue homogenate. The test has a diagnostic sensitivity of 70 and 45%, respectively, compared to single-step and nested PCR. The diagnostic specificity of the test was 82%. This test is a simple and rapid on-farm test which can be used to corroborate clinical signs for the detection of WSSV in grow-out ponds.

Studies on radiation pasteurisation of medium fatty fish 2. Precursors of yellow discolouration in irradiated white pomfret (Stromateus cinereus)

Triglycerides, phospholipids and sarcoplasmic proteins fractions of white pomfret produced considerable amounts of thiobarbituric acid reactive substances (TBRS) on irradiation. Incubation of malonaldehyde with pomfret skin under aseptic conditions developed yellow pigmentation of the skin tissues, similar in spectral characteristics to those produced on irradiation of the skin.