Development, validation, and utilization of a competitive enzyme-linked immunosorbent assay for the detection of antibodies against Brucella species in marine mammals

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal (Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal (Monachus schauinslandi; n = 28) and bottlenose dolphin (Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.

An investigation of survey technologies and modeling techniques for improving deepwater surveys of nonindigenous species in the Northwestern Hawaiian Islands

Nonindigenous species (NIS) are a major threat to marine ecosystems, with possible dramatic effects on biodiversity, biological productivity, habitat structure and fisheries. The Papahānaumokuākea Marine National Monument (PMNM) has taken active steps to mitigate the threats of NIS in Northwestern Hawaiian Islands (NWHI). Of particular concern are the 13 NIS already detected in NWHI and two invasive species found among the main Hawaiian Islands, snowflake coral (Carijoa riseii) and a red alga (Hypnea musciformis). Much of the information regarding NIS in NWHI has been collected or informed by surveys using conventional SCUBA or fishing gear. These technologies have significant drawbacks. SCUBA is generally constrained to depths shallower than 40 m and several NIS of concern have been detected well below this limit (e.g., L. kasmira – 256 m) and fishing gear is highly selective. Consequently, not all habitats or species can be properly represented. Effective management of NIS requires knowledge of their spatial distribution and abundance over their entire range. Surveys which provide this requisite information can be expensive, especially in the marine environment and even more so in deepwater. Technologies which minimize costs, increase the probability of detection and are capable of satisfying multiple objectives simultaneously are desired. This report examines survey technologies, with a focus on towed camera systems (TCSs), and modeling techniques which can increase NIS detection and sampling efficiency in deepwater habitats of NWHI; thus filling a critical data gap in present datasets. A pilot study conducted in 2008 at French Frigate Shoals and Brooks Banks was used to investigate the application of TCSs for surveying NIS in habitats deeper than 40 m. Cost and data quality were assessed. Over 100 hours of video was collected, in which 124 sightings of NIS were made among benthic habitats from 20 to 250 m. Most sightings were of a single cosmopolitan species, Lutjanus kasmira, but Cephalopholis argus, and Lutjanus fulvus, were also detected. The data expand the spatial distributions of observed NIS into deepwater habitats, identify algal plain as an important habitat and complement existing data collected using SCUBA and fishing gear. The technology’s principal drawback was its inability to identify organisms of particular concern, such as Carijoa riseii and Hypnea musciformis due to inadequate camera resolution and inability to thoroughly inspect sites. To solve this issue we recommend incorporating high-resolution cameras into TCSs, or using alternative technologies, such as technical SCUBA diving or remotely operated vehicles, in place of TCSs. We compared several different survey technologies by cost and their ability to detect NIS and these results are summarized in Table 3.

Survey protocol for invasive species

This protocol was developed by the Biogeography Branch of NOAA’s Center for Coastal Monitoring and Assessment to support invasive species research by the Papahānaumokuākea Marine National Monument. The protocol’s objective is to detect Carijoa riisei and Hypnea musciformis in deepwater habitats using visual surveys by technical divers. Note: This protocol is designed to detect the presence or absence of invasive species. A distinct protocol is required to collect information on abundance and impact, or monitor changes over time.

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.