Sighting patterns reveal unobserved pupping events to revise reproductive rate estimates for Hawaiian monk seals in the main Hawaiian Islands

We used sighting reports, including decades of citizen-reported Hawaiian monk seal (Neomonachus schauinslandi) sightings, to describe female breeding biology and reproductive success in the main Hawaiian Islands. We first used this data set to describe the timing of events in the female reproductive cycle. We then conducted an expert review of patterns in sighting histories to detect unobserved pupping events. Finally, we estimated the age-specific reproductive curve for female monk seals in the main Hawaiian Islands. Charting reproductive cycles showed indications of the robust condition of female monk seals in the main Hawaiian Islands; they nursed pups 12% longer than their counterparts in the Northwestern Hawaiian Islands and regained condition to molt more quickly after weaning a pup. By examining sighting histories, we were able to infer 25 unobserved pupping events that had previously gone uncounted. We accounted for additional uncertainty with a randomization procedure. After accounting for unobserved pupping events, the age-specific reproductive rate of main Hawaiian Islands monk seals exceeded 0.70 for prime aged females (8–18 years). This is the highest reproductive rate reported for any of the Hawaiian monk seal breeding sites, illustrating the important role of the main Hawaiian Islands population in Hawaiian monk seal recovery.     

Infectious disease monitoring of the endangered Hawaiian monk seal

As part of conservation efforts between 1997 and 2001, more than 25% (332 animals) of the endangered Hawaiian monk seal (Monachus schauinslandi) population was sampled in the northwestern Hawaiian Islands. Serum samples were tested for antibodies to viruses, bacteria, and parasites known to cause morbidity and mortality in other marine mammal species. Antibodies were found to phocine herpesvirus-1 by using an enzyme-linked immunosorbent assay, but seropositive results were not confirmed by virus neutralization test. Antibodies to Leptospira bratislava, L. hardjo, L. icterohaemorrhagiae, and L. pomona were detected in seals from several sites with the microagglutination test. Antibodies to Brucella spp. were detected using 10 conventional serologic tests, but because of inconsistencies in test results and laboratories used, and the lack of validation by culture, the Brucella serology should be interpreted with caution. Antibodies to B. canis were not detected by card test. Chlamydophila abortus antibodies were detected by complement fixation (CF) test, and prevalence increased significantly as a function of age; the low sensitivity and specificity associated with the CF make interpretation of results difficult. Antibodies to Toxoplasma gondii and Dirofilaria immitis were rarely found. There was no serologic evidence of exposure to four morbilliviruses, influenza A virus, canine adenovirus, caliciviruses, or other selected viruses. Continuous surveillance provides a means to detect the introduction or emergence of these or other infectious diseases, but it is dependent on the development or improvement of diagnostic tools. Continued and improved surveillance are both needed as part of future conservation efforts of Hawaiian monk seals.     

Estimating the carrying capacity of French Frigate Shoals for the endangered Hawaiian monk seal using Ecopath with Ecosim

The carrying capacity of the French Frigate Shoals (FFS) region for the endangered Hawaiian monk seal was appraised using an updated version of the original FFS Ecopath model ( Polovina 1984 ). Model parameters were updated using recent literature, and data from surveys of the seal population and its bottom‐associated prey. Together they produced a static mass balance model for 1998 when the prey surveys began. The Ecopath‐estimated monk seal biomass was 0.0045 t/km², which was in close agreement with the biomass calculated from monk seal field beach counts (0.0046 t/km²). Model simulations through time were done in Ecosim using the Ecopath balanced model and included fisheries data time series from 1998 to 2008. Monk seal biomass declined concurrently with decreases in benthic bottomfish biomass, which were influenced by large‐scale changes in the environment of the North Pacific. This model scenario was extended from 2010, when the last permitted fishery in the Northwestern Hawaiian Islands was closed, through to 2040, assuming a constant environmental signal. Model results for this time period did not show a recovery of monk seals that exceeded the initial 1998 model biomass levels, highlighting the importance of including environmental variability in estimates of monk seals recovery at FFS.     

Toxoplasmosis in an elephant seal (Mirounga angustirostris)

Toxoplasma gondii infections in fish-eating marine mammals is intriguing and indicative of contamination of the sea environment with oocysts. Toxoplasma gondii was identified in an elephant seal (Mirounga angustirostris) that had encephalitis. Tissue cysts were found in sections of cerebrum, and the diagnosis was confirmed by immunohistochemical staining with T. gondii-specific polyclonal rabbit serum. This is the first report of T. gondii infection in an elephant seal.     

Seasonal testosterone pattern in Hawaiian monk seals (Monachus schauinslandi).

Blood samples from four captive male Hawaiian monk seals were collected at intervals of one month for one year for testosterone assay. Plasma testosterone concentrations, measured by radioimmunoassay, revealed a clear seasonal pattern. The lowest mean testosterone concentration (0.09 +/- 0.04 ng ml-1) occurred in January, and the highest (1.78 +/- 0.40 ng ml-1) in June. The seasonal occurrence of births and of injuries related to mating in wild populations of Hawaiian monk seals showed a distinct association with the period of high testosterone. This study supports other data that indicate that the Hawaiian monk seal is a seasonal breeder and is reproductively active for longer than monachine seals that live in higher latitudes.     

CONCENTRATIONS OF ESTRONE SULFATE and PROGESTERONE IN PLASMA and SALIVA, VAGINAL CYTOLOGY, and BIOELECTRIC IMPEDANCE DURING THE ESTROUS CYCLE OF THE HAWAIIAN MONK SEAL (MONACHUS SCHAUNSLAnDI)

Blood and saliva samples, vaginal swabs, and bioelectric impedance measurements were collected 1–3 times per week from a captive adult female Hawaiian monk seal ( Monacbus schauinslandi ) during the spring and summer of 1991. During 1992 saliva only was collected on average 4 times weekly. Concentrations of progesterone and estrone sulfate, and the periodic appearance of cornified epithelial cells from the vagina indicated consecutive estrous cycles ranging 35 ± 3 days in duration. Progesterone concentrations in plasma and saliva had a correlation of 0.868, while estrone sulfate had a correlation of 0.982. Bioimpedance of the whole seal body resulted in a pattern similar to the estrone sulfate concentrations, but shifted forward by 2 d. Bioimpedance of the upper vaginal tissues paralleled the estrone sulfate pattern while the bioimpedance of the lower reproductive tract fluctuated without a distinct pattern. The luteal phase ranged 17–20 d and the follicular phase was 15–18 d in length. These results indicate that: (1) the Hawaiian monk seal, in captivity, is a polyestrous seal, in contrast to other phocid seals that have been studied; (2) salivary concentrations of estrogen and progesterone may provide an accurate, less-invasive method of monitoring reproductive hormones in captive Hawaiian monk seals; and (3) vaginal cytology and bioelectric impedance reflect physiological changes associated with the estrous cycle.     

Biogeography and taxonomy of extinct and endangered monk seals illuminated by ancient DNA and skull morphology

Extinctions and declines of large marine vertebrates have major ecological impacts and are of critical concern in marine environments. The Caribbean monk seal, Monachus tropicalis, last definitively reported in 1952, was one of the few marine mammal species to become extinct in historical times. Despite its importance for understanding the evolutionary biogeography of southern phocids, the relationships of M. tropicalis to the two living species of critically endangered monk seals have not been resolved. In this study we present the first molecular data for M. tropicalis, derived from museum skins. Phylogenetic analysis of cytochrome b sequences indicates that M. tropicalis was more closely related to the Hawaiian rather than the Mediterranean monk seal. Divergence time estimation implicates the formation of the Panamanian Isthmus in the speciation of Caribbean and Hawaiian monk seals. Molecular, morphological and temporal divergence between the Mediterranean and “New World monk seals” (Hawaiian and Caribbean) is profound, equivalent to or greater than between sister genera of phocids. As a result, we classify the Caribbean and Hawaiian monk seals together in a newly erected genus, Neomonachus. The two genera of extant monk seals (Monachus and Neomonachus) represent old evolutionary lineages each represented by a single critically endangered species, both warranting continuing and concerted conservation attention and investment if they are to avoid the fate of their Caribbean relative.     

Experimental Toxoplasma gondii infection in grey seals (Halichoerus grypus)

Laboratory-reared animals were used to assess the susceptibility of seals (Halichoerus grypus) to Toxoplasma gondii infection. Four seals were each orally inoculated with 100 or 10,000 oocysts of T. gondii (VEG strain), and another 4 seals served as negative controls. Occasionally, mild behavioral changes were observed in all inoculated seals but not in control animals. A modified agglutination test revealed the presence of antibodies to T. gondii in sera collected from inoculated seals and mice inoculated as controls. No evidence of the parasite was found on an extensive histological examination of seal tissues, and immunohistochemical staining of tissue sections from inoculated seals revealed a single tissue cyst in only 1 seal. Control mice inoculated with 10 oocysts from the same inoculum given to seals became serologically and histologically positive for T. gondii. Cats that were fed brain or muscle tissue collected from inoculated seals passed T. gondii oocysts in feces. This study demonstrates that T. gondii oocysts can establish viable infection in seals and supports the hypothesis that toxoplasmosis in marine mammals can be acquired from oocysts in surface water runoff and sewer discharge.     

Nutrient, fatty acid, amino acid and mineral analysis of natural prey of the Hawaiian monk seal, Monachus schauinslandi.

Proximate nutrients, gross energy content, mineral, amino acid and fatty acid composition were determined for teleost, cephalopod and crustacean prey of the Hawaiian monk seal. Crude protein was highest in the octopus, Octopus cyanea (80.0%), crude fat was highest in the Muraenid teleost, Gymnothorax eurostus (14.1%), whereas crude ash was highest in the lobster, Panulirus marginatus (11.6%). Gross energies ranged from 4.0 +/- 0.01 kcal g-1 in the Labrid teleost Bodianus bilulunatus to 6.0 +/- 0.12 kcal g-1 in the moray eel, Gymnothorax undulatus. Essential amino acids occurred in lower concentrations as a percentage of the total amino acids (35.8 +/- 2.6%) than non-essential amino acids (64.2 +/- 2.6%), but the ratio of individual amino acids to total amino acid concentrations were similar to those required by some monogastric terrestrial species and fingerling salmon. The fatty acid concentrations varied widely among species (range = 1.2-16.5 mg 100 mg-1); however, the teleosts had higher total fatty acids than the non-teleosts. This study indicates that, from a nutritional standpoint, some prey may be more beneficial to the Hawaiian monk seal; however, these prey are not necessarily the most abundant or available to some populations of the monk seal.     

Hepatitis associated with a Sarcocystis canis-like protozoan in a Hawaiian monk seal (Monachus schauinslandi)

A Hawaiian monk seal (Monachus schauinslandi) died in captivity at the National Marine Fisheries Service, Kewalo Basin Facility in Honolulu, Hawaii. The animal was icteric, and the liver was friable. Microscopic lesions were detected in the colon and liver. Colonic lesions included multifocal, necrohemorrhagic colitis associated with gram-negative bacilli. The liver lesions included random hepatic necrosis and cholestasis. Asexual stages of a Sarcocystis canis-like apicomplexan were detected in hepatocytes. The parasite divided by endopolygeny. Merozoites occasionally formed rosettes around a central residual body. Ultrastructurally, merozoites lacked rhoptries. This is the first report of S. canis infection in M. schauinslandi, which is an endangered pinniped in U.S. waters.     

REPRODUCTIVE PATTERNS OF THE HAWAIIAN MONK SEAL

We evaluated reproductive patterns of the Hawaiian monk seal ( Monachus schauinslandi ) using a combination of fitted age-specific reproductive curves and analysis of reproductive patterns of individual females. We review the difficulties inherent in the acquisition and modeling of reproductive data with emphasis on the significance of reproductive senescence to populations with dissimilar age/sex compositions. Validation of the fitted reproductive parameters was accomplished by Monte Carlo sampling of parameter distributions to compare the expected number of pups with the observed production. Although the fitted reproductive functions appear to provide an acceptable fit to the raw reproductive data, we found that the fitted curves did a poor job of predicting the actual pup production in individual years because of high variability among years. To further verify, and elaborate on, the patterns in the pooled (multi-seal, and multi-year) rates, we examined attributes of the reproductive performance of individual seals. The attributes included age of primiparity, reproductive rates computed over several age ranges, and the relationship between reproductive performance and seal longevity. Analysis of individual seal patterns reinforced the conclusion that reproductive senescence is operative in monk seal populations.     

IDENTIFYING DIURNAL FORAGING HABITAT OF ENDANGERED HAWAIIAN MONK SEALS USING A SEAL-MOUNTED VIDEO CAMERA

The Hawaiian monk seal ( Monachus schauinslandi ) is thought to be a foraging generalist, preying on numerous species in diverse habitats of the subtropical Northwestern Hawaiian Islands. At the atoll of French Frigate Shoals, recent evidence of emaciation and low survival in monk seals prompted a search for their specific prey communities and foraging habitat.
A video camera (National Geographic Television's CRITTERCAM) fitted to 24 adult male seals documented benthic and demersal foraging on the deep slopes (50-80 m) of the atoll and neighboring banks. The number of bottom searches for prey was compared by year, time of day, type of bottom, individual seal, and length of bottom time. Analysis of variance identified a significant interaction of seal and bottom type, explaining 65% of the total variance. Seals fed on communities of cryptic fauna (fish and large invertebrates) in transitional "ecotone" regions of low relief where consolidated substrate, rubble, and talus bordered areas of sand. Independent areal surveys of bottom types throughout the atoll and neighboring banks suggest that the type of bottom selected as foraging habitat represents a relatively small percentage of the total benthic area available.     

Do monk seals exert top‐down pressure in subphotic ecosystems?

Patterns of subphotic fish assemblages on seamounts in the Northwestern Hawaiian Islands were identified and compared for potential structuring influences, including the bottom-up effects of regional oceanic productivity and top-down predation pressure exerted by visiting monk seals ( Monachus schauinslandi ). Patterns in fish size, density, and biomass were evaluated at the deep extreme (350–500 m) of the seals feeding range to avoid confounding effects of diverse shallow habitats ( e.g. , coral reefs). Fish number and size were used to calculate biomass density of the seamount fish assemblages that were then compared to the independent variables of summit depth, substrate type, relief, oceanic productivity, distance to seal colonies, and seal colony population. Only the variables of distance to seal colony and seal colony population were retained in a multiple regression model that explained 31% of the variance. Despite the presence of obvious regional differences in oceanic productivity, the overall patterns in the subphotic fish assemblages are better explained by the top-down hypothesis of predation pressure from monk seals.     

EFFECTS OF RESEARCH HANDLING ON THE ENDANGERED HAWAIIAN MONK SEAL

We examined the effects of research handling on free-ranging endangered Hawaiian monk seals, Monachus schauinslandi , by analyzing differences in subsequent year survival, migration, and condition between handled seals and controls during 1983–1998. Each of 549 handled seals was matched to a control seal of the same age, sex, location, and year. Handling included instrumentation with tel metry devices ( n = 93), blood sampling ( n = 19), and tagging ( n = 437). No significant differences were found between handled seals and their controls in one-year resighting rates, observed migration rates, or condition. Resighting rates of handled and control seals were high (80%-100%). Available sample sizes were sufficient to detect reasonably small (9%-20%) differences in resighting rates had they existed among instrumented or tagged seals and controls (α= 0.05, power = 0.90). Too few seals were captured for blood sampling to detect even large differences in their resighting rates. However, blood samples were drawn from most instrumented seals, and there was no indication that this larger group suffered harmful effects. Duration of restraint during flipper tagging had no effect on subsequent probability of resighting. Our analysis suggests that conservative selection procedures and careful handling techniques have no deleterious effects on Hawaiian monk seals.     

Meningoencephalitis associated with an unidentified apicomplexan protozoan in a Pacific harbor seal

A Pacific harbor seal (Phoca vitulina richardsii) was found on the central California coast with neurologic signs and labored breathing, which were unresponsive to treatment. Necropsy revealed a nonsuppurative necrotizing meningoencephalitis, a multilocular thymic cyst, and nonsuppurative cystitis and renal pyelitis. Microscopic examination revealed protozoans in the brain, thymic cyst, and bladder mucosa. Ultrastructurally, the protozoal tachyzoites were different from those of Neospora caninum, Toxoplasma gondii, and Sarcocystis neurona; the rhoptries were small and had electron-dense contents, and the organism divided by endodyogeny. Specific antibodies were not detected in serum using agglutination (N. caninum, T. gondii) and immunoblot assays (S. neurona). Immunohistochemistry for these organisms was negative. Polymerase chain reaction on brain tissue using specific primers did not amplify T. gondii deoxyribonucleic acid. The meningoencephalitis in this seal thus appears to have been caused by a novel protozoan.     

A COPROLOGICAL SURVEY OF HELMINTH PARASITES OF THE HAWAIIAN MONK SEAL FROM THE NORTHWESTERN HAWAIIAN ISLANDS

In conjunction with an Hawaiian monk seal, Monachus schauinslandi , breeding enhancement program, fecal samples were examined for the presence of helminth parasite products (eggs, larvae, and proglottids). Eighty-two samples were examined from four island groups in the northwestern Hawaiian Islands: 38 from Kure Atoll, 32 from French Frigate Shoals, 11 from Laysan Island, and one from Lisianski Island. Identified helminth products include those of Diphyllobothrium sp., D. cameroni, Contracaecum turgidum, Anisakis sp., Corynosoma rauschi , and Trematoda. In addition, eggs from Halarachne laysanae (Atari) were found in 3 samples. Eggs of C. turgidum were present in the highest percentage (>64%) of samples from all locations. Eggs of Anisakis sp. and Trematoda are reported from this host for the first time.     

Viral susceptibility of newly established cell lines from the Hawaiian monk seal Monachus schauinslandi

Ten of 11 cell lines, recently established from the snout (MS-SN), periorbital soft tissue (MS-EY), liver (MS-LV), kidney (MS-KD), lung (MS-LG), spleen (MS-SP), heart (MS-HT), thyroid (MS-TY), brain (MS-BR) and urinary bladder (MS-UB) of a juvenile Hawaiian monk seal Monachus schauinslandi, were evaluated in vitro for their susceptibility to 5 mammalian viruses: herpes simplex virus type 1 (HSV-1), vesicular stomatitis virus (VSV), reovirus type 3 (Reo-3), poliovirus type 1 (Polio-1) and vaccinia virus (Vac); 5 fish viruses: channel catfish herpesvirus (CCV), infectious hematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV), fish rhabdovirus carpio (RC) and viral hemorrhagic septicemia virus (VHSV); and 2 marine mammal morbilliviruses: phocine distemper virus (PDV) and dolphin distemper virus (DMV). Four well-established continuous cell-lines of nonhuman primate (Vero) and fish (EPC, CHSE-214 and BB) origin served as controls to standardize the virus infectivity assays. Virus yields were quantified as 50% tissue culture infectious dose (TCID50) ml(-1) on Day 7 post-inoculation. Results of the viral challenge assays revealed that the monk seal cell lines shared a similar pattern of susceptibility to the mammalian viruses. Despite their different tissue origins, all monk seal cells were sensitive to HSV-1, Vac, VSV and Reo-3, but were refractory to Polio-1. A characteristic viral-induced cytopathic effect was noted with VSV and Reo-3, and distinct plaques were observed for HSV-1 and Vac. Monk seal cell lines were also susceptible to PDV and DMV, 2 morbilliviruses isolated from seals and dolphins, respectively. By contrast, these cell lines were generally resistant to VHSV, IHNV and IPNV, with varying susceptibility to RC and CCV. The wide range of viral susceptibility of these monk seal cell lines suggests their potential value in studying viruses of monk seals and other marine mammals.     

Dramatic shifts in Hawaiian monk seal distribution predicted from divergent regional trends

Total estimated abundance of Hawaiian monk seals was just 1,161 individuals in 2008 and this number is decreasing. Most monk seals reside in the remote Northwestern Hawaiian Islands (NWHI) where the decline is approximately 4%/yr, whereas relatively fewer seals currently occupy the main Hawaiian Islands (MHI). It is widely accepted that the MHI population is increasing, although there are no formal estimates of total abundance, population growth rate or vital rates. This lack of information has hampered efforts to anticipate future scenarios and plan conservation measures. We present the first estimates of MHI monk seal survival and age‐specific reproductive rates. Using these rates, a conservative estimate of current MHI abundance and a previously published stochastic simulation model, we estimate the MHI population growth rate and projected abundance trend. Analogous estimates for the NWHI are derived from a much richer data set. Estimated survival from weaning to age 1 yr is 77% in the MHI, much higher than recent NWHI estimates ranging from 42% to 57%. Moreover, MHI females begin reproducing at a younger age and attain higher birth rates than observed in the NWHI. The estimated MHI intrinsic rate of population growth is 1.07 compared to a 0.89–0.96 range in the NWHI. Assuming an initial abundance of 152 animals in the MHI, projections indicate that if current demographic trends continue, abundance in the NWHI and MHI will equalize in approximately 15 yr. These results underscore the imperative to mitigate the NWHI decline while devoting conservation efforts to foster population growth in the MHI, where documented threats including fishery interactions, direct killing, and disease could rapidly undo the current fragile positive trend.     

Seroprevalence of Toxoplasma gondii antibodies in wild dolphins from the Spanish Mediterranean coast

Although Toxoplasma gondii infection has been found occasionally in cetaceans, little is known of the prevalence of antibodies to T. gondii in wild dolphins. Antibodies to T. gondii were determined in serum samples from 58 dolphins stranded in the Spanish Mediterranean coast. Modified agglutination test was used to determine T. gondii antibodies, and a titer of 1:25 was considered indicative of T. gondii infection. Antibodies to T. gondii were found in 4 of 36 striped dolphins (Stenella coeruleoalba), in 2 of 4 common dolphins (Delphinus delphis), in 4 of 7 bottlenose dolphins (Tursiops truncatus), and in 1 harbour porpoise (Phocoena phocoena). Antibodies were not found in 9 Risso's dolphins (Grampus griseus) and in 1 long-finned pilot whale (Globicephala melas) surveyed. The results indicate that T. gondii infection is frequent in at least 3 dolphin species from the Mediterranean Sea.