Toxoplasma gondii antibodies in tropical seabirds from the Rocas Atoll Biological Reserve, Brazil.

**Abstract:** Rocas Atoll (3°52′S 33°49′W) is the only atoll in the South Atlantic Ocean, located approximately 260 km off the Northeast coast of Brazil. Despite its small surface (two islets with a combined area of 0.36 km2), Rocas Atoll is home to approximately 150,000 seabirds. Five species breed on the atoll: Black Noddy (Anous minutus), Brown Noddy (Anous stolidus), Sooty Tern (Onychoprion fuscatus), Masked Booby (Sula dactylatra), and Brown Booby (Sula leucogaster). Additionally, small numbers of Red-footed Booby (Sula sula) and Magnificent Frigatebird (Fregata magnificens) also roost on the islets. In June 2017, blood was collected from the tarsus (Suliformes) or jugular vein (Charadriiformes) of 214 seabirds: 26 A. minutus, 42 A. stolidus, 46 O. fuscatus, 33 S. dactylatra, 27 S. leucogaster, 20 S. sula, and 20 F. magnificens. Thin blood smears were freshly prepared, fixed with methanol, stained with Rosenfeld-Giemsa, and examined under the microscope. Two blood parasites were detected: Babesia sp. in Brown Booby (prevalence = 44%) and Haemoproteus sp. in Magnificent Frigatebird (prevalence = 10%). Although a previous study had detected DNA of Haemoproteus sp. in the blood of Brown Noddy, this parasite was not seen in this species' blood smears. Leukocyte profiles were generally similar across species and age groups, with the following averages (mean ± S.D.): heterophils 40.3%±15.9%, lymphocytes 53.8%±15.9%, monocytes 4.2%±3.9%, eosinophils 1.7%±3.0%, and basophils 0%±0.1%. The proportion of eosinophils was highest in S. dactylatra and S. leucogaster, especially in adults, suggesting a higher exposure to helminth parasites. The heterophil-to-leucocyte ratio, an indicative of physiological stress, was higher in adults than in chicks and immatures in most species, with the exception of A. stolidus where an opposite pattern was noted. This is the first hematological study of seabirds at Rocas Atoll, and is part of a broader health survey program that aims to identify pathogens and health challenges that may bear significance for seabird conservation at Brazil's oceanic islands. **Authors:** Ralph Vanstreels¹, Daniela Mariani², Renata Hurtado¹, Veridiana Galizia³, Maurizélia Silva⁴, Jean Ramos Silva² ¹Institute of Research and Rehabilitation of Marine Animals, ²Universidade Federal Rural de Pernambuco, ³Autonomous veterinarian, ⁴Instituto Chico Mendes de Conservação da Biodiversidade

This paper reports on post-1991 census data and on the breeding phenology of seabirds of the Chesterfield-Bampton and Bellona groups of coral islets in the Coral Sea. In total, 13 resident bird species were observed [Wedge-tailed Shearwater (Puffinus pacificus), Masked Booby (S. dactylatra), Brown Booby (Sula leucogaster), Red-footed Booby (S. sula), Lesser Frigatebird (Fregata ariel), Great Frigatebird (F. minor), Black Noddy (Anous minutus), Brown Noddy (A. stolidus), Crested Tern (Sterna bergii), Sooty Tern (S. fuscata), Fairy Tern (S. nereis), Black-naped Tern (S. sumatrana), and Buff-banded Rail (Gallirallus philippensis)]. Segregation for nesting habitat was similar to that previously observed on other coral-reef islets of the Coral Sea. Breeding periods were either in the winter (Masked and Red-footed Boobies, Frigatebirds, Fairy Tern) or in the summer (Wedge-tailed Shearwater, Black and Brown Noddies, Crested and Black-naped Terns) or year-round (Brown Booby). Sooty Terns bred twice a year (summer and spring), but this was not consistent across years. Estimates of breeding population sizes for the whole Chesterfield-Bampton and Bellona groups are proposed for Wedge-tailed Shearwater (90,000 to 106,000 breeding pairs), Masked Booby (280–500 pairs), Brown Booby (3,800–5,800 pairs), Redfooted Booby (7,200–7,300 pairs), Lesser Frigatebird (1,600 pairs), Great Frigatebird (350–480 pairs), Black Noddy (29,000–45,000 pairs), Brown Noddy (15,000–23,000 pairs), Crested Tern (80–100 pairs), Sooty Tern (11,000–46,000 pairs), and Black-naped Tern (70–90 pairs). Interannual fluctuation in breeding population size was apparent in Wedge-tailed Shearwater. Over the last 30 yr, an increase in Brown Booby abundance was noted, whereas declines are suspected for the Fairy Tern and Buff-banded Rail. Among the threats to nesting seabirds are stress and other disturbances caused by human frequentation, including poaching of seabird chicks and introduced mice.

Seroprevalence of Toxoplasma gondii in seabirds from Abrolhos Archipelago, Brazil

Persistent organic pollutant patterns in seabirds from marine protected areas in the tropical Atlantic Ocean

**Abstract:** The islets of the Tubbataha Reefs Natural Park serve as an important breeding ground for seabirds. This study aimed to determine how the variability in climate has influenced the population dynamics and phenology of six resident breeding seabird species in the park: Red-footed Booby, Brown Booby, Great Crested Tern, Sooty Tern, Brown Noddy and Black Noddy. Survey methods followed Bibby et al. (2002). The data suggests that the increase in sea surface temperature in the area did not negatively influence the inter-annual variability of the seabird population. However, during El Niño events the effects of sea surface temperature changes to Great Crested Tern, Sooty Tern, and Brown Noddy were significant. Chlorophyll a concentration, which was used as a proxy for prey availability, could not be linked to the presence of adult birds, but it reliably predicted the breeding phenology of Great Crested Tern and Sooty Tern in the area. The peak of sea surface temperature was strongly linked to the start of egg-laying for the Great Crested Tern. The future changes in climate could possibly influence these environmental factors and consequently affect the phenology as well as the population of the seabird species in the area. The future occurrence of long and strong El Niño episodes might delay the breeding of these seabirds. The result of this and further studies will help park management better understand seabirds and come up with effective conservation strategies for the species. **Authors:** Maria Retchie Pagliawan-Alaba¹, Angelique Songco¹, Arnold van Vliet², Arne Jensen³, Rowell Alarcon¹, Gerlie Gedoria¹ ¹Tubbataha Management Office, ²Wageningen University and Research Centre, ³Wild Bird Club of the Philippines

During the hunting season of 1993, hearts from 28 black bears (Ursus americanus) from Pennsylvania were examined for Toxoplasma gondii infection. Serum samples from heart blood were examined for T. gondii antibodies by the use of the modified agglutination test (MAT), the Sabin-Feldman dye test (DT), the latex agglutination test (LAT), and the indirect hemagglutination test (IHA). One-hundred grams of myocardium from each bear were bioassayed in mice. Approximately 500 g of myocardium from 11 bears were fed to 11 cats, and feces from those cats were examined for T. gondii oocysts. Toxoplasma gondii MAT antibodies (> or = 1:40) were found in 22 bears in titers of 1:40 (4 bears), 1:80 (3 bears), 1:160 (7 bears), 1:320 (4 bears), and > or = 1:2,000 (4 bears). Antibodies to T. gondii by DT (> or = 1:10), LAT (> or = 1:32), and IHA (> or = 1:64) tests were found in 21, 9, and 6 bears, respectively. All 6 bears that did not have T. gondii antibodies in MAT were also negative in DT, IHA, and LAT. Viable T. gondii was isolated from 3 bears by bioassays in mice and from an additional 7 bears by bioassays in cats. All 10 bears that had viable T. gondii had MAT and DT antibodies but only 6 and 5 of them had antibodies by LAT and IHA, respectively.

Bird pathogens promote morbidity and mortality that affect bird population stability. In this context, the present study aimed to detect blood intracellular parasites by molecular methods in four species of seabirds, Masked Booby Sula dactylatra, Red-footed Booby S. sula, Brown Booby S. leucogaster, and Magnificent Frigatebird Fregata magnificens, on the Brazilian coast. Blood samples were collected in the Archipelagos of São Pedro and São Paulo, Fernando de Noronha, Atol das Rocas, and Trindade and Martim Vaz. Genomic DNA was extracted with proteinase K digestion and purification with phenol and chloroform. For each sample, three independent polymerase chain reactions were performed with a positive control for Plasmodium and Haemoproteus. Corroborating other studies, our results indicate a low prevalence of Plasmodium and Haemoproteus parasitemia in this group of birds in the wild. We propose that the low parasitemia occurred due to 1) the absence of the vector for transmission among individuals of these islands; 2) the low number of parasitized individuals detected in the colonies sampled; and 3) an effective immune response that prevented installation of the parasite. Only the Masked Booby and Magnificent Frigatebird have sufficient flight capacity to reach the Brazilian coast, but Masked Boobies rarely visit waters near the coast. Without vector studies at the archipelagos that were sampled, these hypotheses cannot be assessed.

The Great Barrier Reef is an iconic ecosystem, known globally for its rich marine biodiversity that includes many thousands of tropical breeding seabirds. Despite indications of localized declines in some seabird species from as early as the mid‐1990s, trends in seabird populations across the reef have never been quantified. With a long history of human impact and ongoing environmental change, seabirds are likely sentinels in this important ecosystem. Using 4 decades of monitoring data, we estimated site‐specific trends for 9 seabird species from 32 islands and cays across the reef. Trends varied markedly among species and sites, but probable declines occurred at 45% of the 86 species‐by‐site combinations analyzed compared with increases at 14%. For 5 species, we combined site‐specific trends into a multisite trend in scaled abundance, which revealed probable declines of Common Noddy (Anous stolidus), Sooty Tern (Onychoprion fuscatus), and Masked Booby (Sula dactylatra), but no long‐term changes in the 2 most widely distributed species, Greater Crested Tern (Thalasseus bergii) and Brown Booby (Sula leucogaster). For Brown Booby, long‐term stability largely resulted from increases at a single large colony on East Fairfax Island that offset declines at most other sites. Although growth of the Brown Booby population on East Fairfax points to the likely success of habitat restoration on the island, it also highlights a general vulnerability wherein large numbers of some species are concentrated at a small number of key sites. Identifying drivers of variation in population change across species and sites while ensuring long‐term protection of key sites will be essential to securing the future of seabirds on the reef.

The Tinhosas islands, in São Tomé e Príncipe, host the most important seabird breeding colony in the Gulf of Guinea, but information on its conservation status was hitherto unpublished or anecdotal, the last assessment having been performed in 1997. A two-day expedition to the Tinhosas islands was undertaken to estimate the status of breeding seabirds in 2013. Four of the five seabird species known to breed in São Tomé e Príncipe, namely Brown Booby Sula leucogaster, Sooty Tern Onychoprion fuscatus, Brown Noddy Anous stolidus and Black Noddy Anous minutus, occur on the Tinhosas. A decrease of 80% in Brown Booby numbers, possibly due to occasional exploitation, and a 30% increase in Sooty Tern and Black Noddy numbers, were found compared to 1997 data although survey methods differed. Breeding of Brown Noddy and Madeiran Storm-petrel Hydrobates castro remains unconfirmed. Our estimates confirmed that BirdLife International Important Bird and Biodiversity Area criteria are met for at least one species, the Sooty Tern. The islands are not legally protected, nonetheless, apart from moderate levels of disturbance by fishermen who land on Tinhosa Grande, no alien species were seen, and no immediate threats to the Tinhosas colony were detected. Multiple visits within and between years are recommended, to census breeders, monitor threats and establish breeding phenologies.

Toxoplasma gondii is the aetiological agent of the zoonotic disease toxoplasmosis and transmitted among other ways by chemically and physically untreated, that is, raw pork to humans. The detection of Toxoplasma gondii is impossible by currently practiced meat inspection, but serological tests can be used to detect Toxoplasma gondii antibodies in pig herds and can consequently be helpful to identify potentially contaminated pork. Therefore, appropriate serological tests are required. In this study, serum samples of 1368 naturally exposed slaughter pigs from 73 Austrian farms were collected. Serum samples of at least 16 slaughter pigs per farm were tested. The prevalence of Toxoplasma gondii antibodies in serum was measured by a commercial available modified agglutination test (MAT) and compared to three different commercial available enzyme-linked immunosorbent assays (ELISA). The MAT detected 6.5%, ELISA I 6.7%, ELISA II 4.8% and ELISA III 4.3% of the pigs as Toxoplasma gondii antibody positive. The agreement, according to the kappa coefficient (κ), was substantial between the MAT and ELISA I (κ = 0.62), II (κ = 0.64) and III (κ = 0.67). A better agreement was determined between ELISA I and II (κ = 0.715), ELISA I and III (κ = 0.747) and ELISA II and III (κ = 0.865). At least one pig per farm was detected Toxoplasma gondii antibody positive in 17 (23.3%) farms by the MAT, 26 (35.6%) farms by ELISA I, 16 (21.9%) farms by ELISA II and 11 (15.1%) farms by ELISA III. Pig farms with a high number of Toxoplasma gondii antibody-positive pigs or high antibody titres were identified by all of the four used serological tests. Concerning the occurrence of Toxoplasma gondii antibodies in Austrian pig farms, a monitoring and surveillance programme would be reasonable to find high-risk farms.

(1) Ozawa (1966) reported the Brown booby, Sula leucogaster ranged as far north as Kusagaki-jima and Sumisu-to, froming the northernmost limit of its breed-ing range, likely on the latter island, in the North-west Pacific Ocean. After that, observations of the Brown booby and other species around these islands were carried out by two ships, the Umitaka Maru and Seiyo Maru, of the Tokyo University of Fisheries. The Umitaka Maru cruised around Kusagaki-jima in July, 1967 and Tori-shima west of the Danjyo-guntto in August 1967. Sumisu-to was observed by the cruises of the Seiyo Maru in July, 1966 and July, 1967 as well as by the Umitaka Maru in February, 1968. The Umitaka Maru cruised around Sofu-gan lying approximately 40 miles south of Tori-shima, Izu-retto on her way northward to Sumisu-to on February 18, 1968.(2) In total 1669 Brown boobies consisted of 6 individuals and 50 flocks were recorded at sea from 42 miles east of Kusagaki-jima westward to the Islands during the period of 1600-1900 (Sunset time) July 23, 1967. The Brown booby flying towards the Islands (nesting place) increased in percentages and the average number per flock decreased as the island was near and the Sunset closed.The distance (mile), then the Brown booby came in sight without an aid of binocular, in relation to the number of the birds in a flock (n) was estimated: D (mile)=0.7 n 0. 19(3) A total of 5 Brown boobies was recorded around Tori-shima lying 18.5 milesnorth-west of the Danjyo-guntto, west of Kyushu on August 6, 1967.(4) The birds recorded around Sumisu-to, in July 1966 were 80 individuals in total of Sooty tern, Sterna fuscata and Common noddy, Anous stolidus and then no Brown booby. In July 1967, 7 Brown boobies and a total of 64 individuals of Sooty tern and Common noddy were observed. In February 1968, 15 individuals of the Brown booby were sighted at all.(5) No birds were seen around Sofu-gan in February 18, 1968.

Seabirds are often free from blood parasites, and a recent review suggested that phylogenetic, ecological and life-history parameters can determine the prevalence of blood parasites in seabirds. However, there is a lack of data available from many seabird groups, and a larger database is needed to understand prevalence patterns of blood parasites. We used a molecular screening approach to detect parasites of the genera Plasmodium, Haemoproteus, Leucocytozoon and Babesia in five species of two genera of seabirds that breed on Atlantic Ocean islands off Brazil. The observed patterns differed between the two bird genera. Like other Laridae, brown noddy, Anous stolidus adults were infected with Haemoproteus with low prevalence. Masked boobies, Sula dactylatra and brown boobies, Sula leucogaster were infected with Babesia. Of the latter, mainly juveniles were infected. In all species, intensity of infection (i.e. number of infected erythrocytes) was so low that parasites remained undetected in blood smears. This may explain the absence of major effects on the body condition of birds, although infected juvenile masked boobies were lighter than juveniles that were not infected with Babesia. Two tree-nesting species; black noddy, Anous minutus and red-footed booby, Sula sula did not have blood parasites, suggesting that tree-nesting may reduce the exposure to arthropod vectors compared with ground nesting in these species.

In recent decades, several seabird populations have declined globally due to anthropogenic activities. In Brazil, 14 seabird species breed at four oceanic islands and one atoll: the Abrolhos, Fernando de Noronha, and Sao Pedro and Sao Paulo (SPSPA) archipelagos; the Trindade/Martin Vaz Islands; and the Atol das Rocas. Seven species are listed as nationally threatened by extinction. This study aimed to present new information on breeding seabird populations in Brazilian oceanic islands, compile all available data previously published and, when possible, to provide updated information on population estimates from censuses carried out sporadically at different islands between 2006 and 2013. Based on new data and the thorough review provided here, of the 35 seabird breeding populations analysed, 14% were increasing (as Red-billed Tropicbird Phaethon aethereus , Magnificent Frigatebird Fregata magnificens and Brown Noddy Anous stolidus in Abrolhos), 11% were decreasing (as Brown Booby Sula leucogaster in Atol das Rocas and Great Frigatebird Fregata minor in Trindade Island), 23% were stable (as White-tailed Tropicbird Phaethon lepturus in Fernando de Noronha and Brown Noddy and Black Noddy Anous minutus in Sao Pedro and Sao Paulo Archipelago), and the remaining 49% were unknown or not possible to evaluate. The Red-footed Booby ( Sula sula ) is locally extinct in Trindade Island, however there are colonies of only a few individuals of other species, such as the Audubon's Shearwater Puffinus lherminieri and Red- billed Tropicbird in Noronha, Black Noddy in Martin Vaz, and Great ( Fregata ariel trinitatis ) and Lesser ( F. m. nicoll i) Frigatebirds in Trindade, that may become extinct soon. Censuses at distinct periods of the breeding cycles and protocols were highly variable, making temporal comparisons difficult. These results indicate an urgent need for long-term studies to improve the scenario to assess seabird population trends based on comparable methodologies, in order to determine trends in the future.

Simple SummaryToxoplasmosis is a zoonotic disease that has serious consequences for immunocompromised individuals, in particular for the fetus during primary infection in pregnant women. The parasite responsible, Toxoplasma gondii, can infect all warm-blooded animals, and studies on antibodies against the parasite in their blood can give a good estimate of its burden in the environment and the risk for human infections. This is less studied in dogs than in other animal species, especially in northern African countries, despite evidence that they can be a potential source of human contamination. The study reported here compared two different methods to assess the presence of antibodies against T. gondii in a cohort of hunting dogs from northern Algeria, as these dogs are in close contact with wildlife and humans, in both rural and urban environments. The results of this study show that about 37% of hunting dogs are infected by T. gondii and that both tests can be used for this diagnostic purpose. This work also highlights the importance of this kind of study in companion animals to assess infectious risk of T. gondii for human populations.Toxoplasmosis is a zoonotic disease, caused by the protozoan Toxoplasma gondii, affecting most warm-blooded animals. Assessing the seroprevalence of T. gondii in different animal species gives a good estimate of the global circulation of the parasite and the risk for human infections. However, the seroprevalence of T. gondii in dogs is not studied as much as other species, despite their close contact with wildlife and humans in rural or urban environments and evidence that dogs can also be a potential source for human contaminations. A commercial enzyme-inked immunosorbent assay (ELISA) kit to detect anti-T. gondii antibodies in sera of hunting dogs potentially naturally infected, was compared to the modified agglutination test (MAT), used as the reference method. The ELISA presented a sensitivity of 76.5% (CI 95%: 60.0–87.6) and a specificity of 87.7% (CI 95%: 76.7–93.9) and a substantial agreement with the MAT for the detection of canine anti-T. gondii antibodies. Both tests can therefore be used widely for epidemiology studies on T. gondii infections in dogs. With a mean seroprevalence of T. gondii infection in hunting dogs from northern Algeria of 36.8% (CI 95%: 34.9–38.7), this study also highlights the importance of T. gondii seroprevalence studies in companion animals to assess infectious risk for human populations.

The current study describes the chewing lice community associated with seabird populations resident at the São Pedro and São Paulo Archipelago in the Atlantic Ocean in Pernambuco State, Brazil. Adults of three species of seabirds were captured using hand-nets for the collection of biometric data and specimens of ectoparasites. In total, 320 birds were examined (120 Sula leucogaster (Boddaert), Brown Booby; 120 Anous stolidus (L.), --Brown Noddy; and 80 Anous minutus Boie, Black Noddy) of which 95% were infested with 8,625 chewing lice, representing eight species of the genera Actornithophilus, Austromenopon, Eidmanniella, Pectinopygus, Quadraceps, and Saemundssonia. On S. leucogaster, Pectinopygus garbei (Pessôa & Guimarães) was more prevalent and had a mean and median intensity of infestation significantly greater than those recorded for Eidmanniella albescens (Piaget). On the two Noddies, the species of Actornithophilus and Quadraceps were significantly more prevalent and abundant than Austromenopon atrofulvum (Piaget) and Saemundssonia remota Timmermann Most of the louse species had a highly aggregated distribution, with k exponent of the negative binomial distribution ranging from 0.04 to 3.06. A weak but significant correlation was found between the abundance of chewing lice and morphometric variables (body weight, wing, beak, tail, and tarsus lengths). It is possible that high rates of infestation have a negative effect on the morphological characteristics of the hosts, including the health of the plumage. All the lice species collected--except for P. garbei (ex S. leucogaster)--were reported for the first time from Brazilian populations of these seabird species.