Detection and characterization of diverse coccidian protozoa shed by California sea lions.

Tissue-cyst forming coccidia in the family Sarcocystidae are etiologic agents of protozoal encephalitis in marine mammals including the federally listed Southern sea otter (Enhydra lutris). California sea lions (Zalophus californianus), whose coastal habitat overlaps with sea otters, are definitive hosts for coccidian protozoa provisionally named Coccidia A, B and C. While Coccidia A and B have unknown clinical effects on aquatic wildlife hosts, Coccidia C is associated with severe protozoal disease in harbor seals (Phoca vitulina). In this study, we conducted surveillance for protozoal infection and fecal shedding in hospitalized and free-ranging California sea lions on the Pacific Coast and examined oocyst morphology and phenotypic characteristics of isolates via mouse bioassay and cell culture. Coccidia A and B were shed in similar frequency, particularly by yearlings. Oocysts shed by one free-ranging sea lion sampled at Año Nuevo State Park in California were previously unidentified in sea lions and were most similar to coccidia infecting Guadalupe fur seals (Arctocephalus townsendi) diagnosed with protozoal disease in Oregon (USA). Sporulated Coccidia A and B oocysts did not replicate in three strains of mice or in African green monkey kidney cells. However, cultivation experiments revealed that the inoculum of fecally-derived Coccidia A and B oocysts additionally contained organisms with genetic and antigenic similarity to Sarcocystis neurona; despite the absence of detectable free sporocysts in fecal samples by microscopic examination. In addition to the further characterization of Coccidia A and B in free-ranging and hospitalized sea lions, these results provide evidence of a new role for sea lions as putative mechanical vectors of S. neurona, or S. neurona-like species. Future work is needed to clarify the distribution, taxonomical status, and pathogenesis of these parasites in sea lions and other marine mammals that share their the near-shore marine environment.

Procedures and Uterine-Sparing Surgeries for Managing Postpartum Hemorrhage: A Systematic Review.

IMPORTANCE: Postpartum hemorrhage (PPH) is one of the leading causes of maternal mortality and morbidity worldwide. OBJECTIVE: The aim of this study was to systematically review literature on procedures (eg, uterine artery embolization) and uterine-sparing surgeries for PPH. EVIDENCE ACQUISITION: We searched MEDLINE and other databases from 1990 to November 2014. Two reviewers independently evaluated studies against predetermined criteria, extracted data, and assessed study quality and strength of the evidence (confidence in the effect). RESULTS: Twenty-eight small studies addressed 1 or more procedures (19 studies of embolization, 5 of uterine tamponade) or surgeries (5 studies of arterial ligation, 5 of uterine compression sutures). Studies primarily evaluated bleeding control and adverse effects. Rates of hemostasis were typically greater than 60% in studies reporting such data after failure of conservative management. Postprocedure infertility occurred in 0% to 43% of women in a small number of studies that reported these data. Uterine tamponade successfully controlled bleeding in more than 50% of women, with few harms reported. Success rates for ligation and sutures ranged from 36% to 96%; harms included surgical injury, infection, and fertility-associated effects. CONCLUSIONS AND RELEVANCE: A limited body of evidence addresses these interventions for PPH. Median rates of hemostasis ranged from 36% to 98%; however, these data come from few studies with less than 2100 total participants. Harms were not well characterized. Some studies with longer-term follow-up reported infertility in women undergoing embolization. Few adverse events with tamponade, ligation, or sutures were reported. Given the insufficient evidence, clinicians must continue to make individual care decisions based on each woman's clinical situation and available management options.

Barriers to Male Involvement in Antenatal Care in Rural Mozambique.

Low rates of antenatal care (ANC) service uptake limit the potential impact of mother-to-child HIV-prevention strategies. Zambézia province, Mozambique, has one of the lowest proportions of ANC uptake among pregnant women in the country, despite the availability of free services. We sought to identify factors influencing ANC service uptake (including HIV counseling and testing) through qualitative methods. In addition, we encouraged discussion about strategies to improve uptake of services. We conducted 14 focus groups to explore community views on these topics. Based on thematic coding of discourse, two main themes emerged: (a) gender inequality in decision making and responsibility for pregnancy and (b) community beliefs that uptake of ANC services, particularly, if supported by a male partner, reflects a woman's HIV-positive status. Interventions to promote ANC uptake must work to shift cultural norms through male partner participation. Potential strategies to promote male engagement in ANC services are discussed.

Epidemiology and pathology of Toxoplasma gondii in free-ranging California sea lions (Zalophus californianus).

The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, as indicated by antibody. Five confirmed and four suspected cases of T. gondii infection were identified by analysis of 1,152 medical records of necropsied sea lions from 1975-2009. One suspected and two confirmed cases were identified in aborted fetuses from a sea lion rookery. Toxoplasmosis was the primary cause of death in five cases, including the two fetuses. Gross and histopathologic findings in T. gondii-infected sea lions were similar to those reported in other marine mammals. The most common lesions were encephalitis, meningitis, and myocarditis. The antibody prevalence in stranded, free-ranging sea lions for 1998-2009 was 2.5% (±0.03%; IgG titer 640). There was an increase in odds of exposure in sea lions with increasing age, suggesting cumulative risk of exposure and persistent antibody over time. The occurrence of disseminated T. gondii infection in aborted fetuses confirms vertical transmission in sea lions, and the increasing odds of exposure with age is consistent with additional opportunities for horizontal transmission in free-ranging sea lions over time. These data suggest that T. gondii may have two modes of transmission in the sea lion population. Overall, clinical disease was uncommon in our study which, along with low prevalence of T. gondii antibody, suggests substantially less-frequent exposure and lower susceptibility to clinical disease in California sea lions as compared to sympatric southern sea otters (Enhydra lutris nereis).

Circulating Fat-Soluble Vitamin Concentrations and Nutrient Composition of Aquatic Prey Eaten by American Oystercatchers (Haematopus palliatus palliatus) in the Southeastern United States.

The American oystercatcher (Haematopus palliatus palliatus) is currently listed as a species of high concern by the United States Shorebird Conservation Plan. Because nutritional status directly impacts overall health and reproduction of individuals and populations, adequate management of a wildlife population requires intimate knowledge of a species' diet and nutrient requirements. Fat-soluble vitamin concentrations in blood plasma obtained from American oystercatchers and proximate, vitamin, and mineral composition of various oystercatcher prey species were determined as baseline data to assess nutritional status and nutrient supply. Bird and prey species samples were collected from the Cape Romain region, South Carolina, USA, and the Altamaha River delta islands, Georgia, USA, where breeding populations appear relatively stable in recent years. Vitamin A levels in blood samples were higher than ranges reported as normal for domestic avian species, and vitamin D concentrations were lower than anticipated based on values observed in poultry. Vitamin E levels were within ranges previously reported for avian groups with broadly similar feeding niches such as herons, gulls, and terns (eg, aquatic/estuarine/marine). Prey species (oysters, mussels, clams, blood arks [Anadara ovalis], whelks [ Busycon carica ], false angel wings [ Petricola pholadiformis ]) were similar in water content to vertebrate prey, moderate to high in protein, and moderate to low in crude fat. Ash and macronutrient concentrations in prey species were high compared with requirements of carnivores or avian species. Prey items analyzed appear to meet nutritional requirements for oystercatchers, as estimated by extrapolation from domestic carnivores and poultry species; excesses, imbalances, and toxicities-particularly of minerals and fat-soluble vitamins-may warrant further investigation.

Diagnosis and treatment of Sarcocystis neurona-induced myositis in a free-ranging California sea lion.

CASE DESCRIPTION: An underweight, lethargic adult female California sea lion (Zalophus californianus) became stranded along the California shore and was captured and transported to a rehabilitation hospital for assessment and care. CLINICAL FINDINGS: Initial physical assessment revealed the sea lion was lethargic and in poor body condition. Active myositis was diagnosed on the basis of concurrent elevations in activities of alanine aminotransferase and creatine kinase detected during serum biochemical analysis. Infection with Sarcocystis neurona was diagnosed after serologic titers increased 4-fold over a 3-week period. Diagnosis was confirmed on the basis of histopathologic findings, positive results on immunohistochemical staining, and results of quantitative PCR assay on biopsy specimens obtained from the diaphragm and muscles of the dorsal cervical region. TREATMENT AND OUTCOME: Anticoccidial treatment was instituted with ponazuril (10 mg/kg [4.5 mg/lb], PO, q 24 h) and continued for 28 days. Prednisone (0.2 mg/kg [0.09 mg/lb], PO, q 12 h) was administered for 2 days and then every 24 hours for 5 days to treat associated inflammation. At the end of treatment, the sea lion was clinically normal, alanine aminotransferase and creatine kinase values were within reference limits, and antibody titers against S neurona had decreased 6-fold. The sea lion was released approximately 3 months after becoming stranded. CLINICAL RELEVANCE: S neurona-induced myositis was diagnosed in a free-ranging California sea lion. On the basis of the successful treatment and release of this sea lion, anticoccidial treatment should be considered for marine mammals in which protozoal disease is diagnosed.

Identification of two novel coccidian species shed by California sea lions (Zalophus californianus).

Routine fecal examination revealed novel coccidian oocysts in asymptomatic California sea lions (Zalophus californianus) in a rehabilitation facility. Coccidian oocysts were observed in fecal samples collected from 15 of 410 California sea lions admitted to The Marine Mammal Center between April 2007 and October 2009. Phylogenetic analysis using the full ITS-1 region, partial small subunit 18S rDNA sequence, and the Apicomplexa rpoB region identified 2 distinct sequence clades, referred to as Coccidia A and Coccidia B, and placed them in the Sarcocystidae, grouped with the tissue-cyst-forming coccidia. Both sequence clades resolved as individual taxa at ITS-1 and rpoB and were most closely related to Neospora caninum. Coccidia A was identified in 11 and Coccidia B in 4 of 12 sea lion oocyst samples successfully sequenced (3 of those sea lions were co-infected with both parasites). Shedding of Coccidia A oocysts was not associated with age class, sex, or stranding location, but yearlings represented the majority of shedders (8/15). This is the first study to use molecular phylogenetics to identify and describe coccidian parasites shed by a marine mammal.

Discovery of three novel coccidian parasites infecting California sea lions (Zalophus californianus), with evidence of sexual replication and interspecies pathogenicity.

Enteric protozoal infection was identified in 5 stranded California sea lions (Zalophus californianus). Microscopically, the apical cytoplasm of distal jejunal enterocytes contained multiple stages of coccidian parasites, including schizonts with merozoites and spherical gametocytes, which were morphologically similar to coccidians. By histopathology, organisms appeared to be confined to the intestine and accompanied by only mild enteritis. Using electron microscopy, both sexual (microgametocytes, macrogamonts) and asexual (schizonts, merozoites) coccidian stages were identified in enterocytes within parasitophorous vacuoles, consistent with apicomplexan development in a definitive host. Serology was negative for tissue cyst-forming coccidians, and immunohistochemistry for Toxoplasma gondii was inconclusive and negative for Neospora caninum and Sarcocystis neurona. Analysis of ITS-1 gene sequences amplified from frozen or formalin-fixed paraffin-embedded intestinal sections identified DNA sequences with closest homology to Neospora sp. (80%); these novel sequences were referred to as belonging to coccidian parasites "A," "B," and "C." Subsequent molecular analyses completed on a neonatal harbor seal (Phoca vitulina) with protozoal lymphadenitis, hepatitis, myocarditis, and encephalitis showed that it was infected with a coccidian parasite bearing the "C" sequence type. Our results indicate that sea lions likely serve as definitive hosts for 3 newly described coccidian parasites, at least 1 of which is pathogenic in a marine mammal intermediate host species.

Persistent organic pollutants in the blood of free-ranging sea otters (Enhydra lutris ssp.) in Alaska and California.

As part of tagging and ecologic research efforts in 1997 and 1998, apparently healthy sea otters of four age-sex classes in six locations in Alaska and three in California were sampled for persistent organic pollutants (POPs) and other chemicals of ecologic or environmental concern (COECs). Published techniques for the detection of POPs (specifically ∑polychlorinated biphenyls [PCBs], ∑DDTs, ∑hexachlorocyclohexanes [HCHs], ∑polycyclic aromatic hydrocarbons [PAHs], ∑chlordanes [CHLs], hexachlorobenzene [HCB], dieldrin, and mirex) in the tissue of dead otters were modified for use with serum from live sea otters. Toxic equivalencies (TEQs) were calculated for POPs with proven bioactivity. Strong location effects were seen for most POPs and COECs; sea otters in California generally showed higher mean concentrations than those in Alaska. Differences in contaminant concentrations were detected among age and sex classes, with high levels frequently observed in subadults. Very high levels of ∑DDT were detected in male sea otters in Elkhorn Slough, California, where strong freshwater outflow from agricultural areas occurs seasonally. All contaminants except mirex differed among Alaskan locations; only ∑DDT, HCB, and chlorpyrifos differed within California. High levels of ∑PCB (particularly larger, more persistent congeners) were detected at two locations in Alaska where associations between elevated PCBs and military activity have been established, while higher PCB levels were found at all three locations in California where no point source of PCBs has been identified. Although POP and COEC concentrations in blood may be less likely to reflect total body burden, concentrations in blood of healthy animals may be more biologically relevant and less influenced by state of nutrition or perimortem factors than other tissues routinely sampled.

Health assessment of American Oystercatchers (Haematopus palliatus palliatus) in Georgia and South Carolina.

The American Oystercatcher (Haematopus palliatus palliatus) is the only species of oystercatcher native to the Atlantic coast of North America and is restricted in distribution to intertidal shellfish beds in coastal areas. Currently, the American Oystercatcher population in South Carolina and Georgia is threatened by widespread habitat loss, resulting in low reproductive success and small population size. Oystercatchers could be an important indicator of ecosystem health because they depend on quality coastal breeding habitat and prey on bivalves, which can accumulate toxins and pathogens from the local environment. Data were collected from American Oystercatchers (n=171) captured at five sites in South Carolina and Georgia between 2001 and 2006. Iridial depigmentation was frequently noted during physical examination and was more prevalent in female birds. Female birds were larger than males on average, but ranges for weight and morphometric measurements had considerable overlap. Mean values were calculated for hematology, plasma biochemistry, and hormone levels, and prevalence of exposure to select pathogens was determined. Mercury was the only trace metal detected in blood samples. These data provide baseline health information needed for longitudinal monitoring and conservation efforts for American Oystercatchers. In addition, this study illustrates the potential use of this species as an indicator for the health of the southeastern US coastal nearshore ecosystem.

Marine species mortality in derelict fishing nets in Puget Sound, WA and the cost/benefits of derelict net removal.

Derelict fishing gear persists for decades and impacts marine species and underwater habitats. Agencies and organizations are removing significant amounts of derelict gear from marine waters in the United States. Using data collected from repeated survey dives on derelict gillnets in Puget Sound, Washington, we estimated the daily catch rate of a given derelict gillnet, and developed a model to predict expected total mortality caused by a given net based on entanglement data collected upon its removal. We also generated a cost:benefit ratio for derelict gear removal utilizing known true costs compared to known market values of the resources benefiting from derelict gear removal. For one study net, we calculated 4368 crab entangled during the impact lifetime of the net, at a loss of 19,656 dollars of Dungeness crab to the commercial fishery, compared to 1358 dollars in costs to remove a given gillnet, yielding a cost:benefit ratio of 1:14.5.