Molecular Detection of Tick-Borne Pathogens in Dogs from Indigenous Communities, Amazon, Brazil.

Background: There are few reports of tick-borne pathogens infecting dogs living in indigenous communities of Brazil. Herein, we aimed to molecularly detect vector-borne pathogens in dogs from two indigenous communities in the Brazilian Amazon. Materials and Methods: We surveyed 327 dogs raised in Amazon region at 2 distinct indigenous ethnicities for the molecular detection of tick-borne pathogens (114 from Tapirapé and 213 from Karajá indigenous ethnicity). Whole blood samples were subjected to PCR and sequencing for Ehrlichia, Babesia, and Hepatozoon. Univariate and multivariate analyses were used to investigate the factors affecting the pathogen infection patterns in dogs. Results: Among the 327 blood samples, 40 were positive for Ehrlichia canis (12.2%), 2 for Anaplasma platys (0.61%), and 204 were positive for Hepatozoon canis (66.5%). Binary Logistic Regression showed association between E. canis infection and ethnicity (p = 0.010) and tick attachment (p = 0.041). Karajá dogs were 3.4 times (95% CI 1.3-8.5) more likely to be positive for E. canis than Tapirapé dogs. Dogs with ticks were 2.5 times more likely (95% CI 1.0-7.6) to be positive for E. canis than dogs without ticks. Conclusions: Our survey expands the knowledge regarding the presence of vector-borne pathogens in dogs from indigenous communities in the Amazon region.

Molecular Characterization of New Haplotype of Genus Sarcocystis in Seabirds from Magdalena Island, Southern Chile.

Evidence of sarcocystid infection was investigated in samples of 16 penguins (Spheniscus. magellanicus), four Dominican gulls (Larus dominicanus) and two Chilean skuas (Stercorarius chilensis) found in Madalenas Islands, Chile, in 2017. Samples of skeletal muscle, cardiac muscle and brain from all birds were screened by a pan-sarcocystid nested-PCR targeting a short fragment of the gene encoding the small ribosomal unit (nPCR-18Sa). The only two positive samples by nPCR-18Sa, both from skuas, were tested by a nested-PCR directed to the internal transcribed spacer 1 (nPCR-ITS1), also a pan-sarcocystidae nested-PCR, and to a nested-PCR directed to the B1 gene (nPCR-B1), for the exclusive detection of Toxoplasma gondii. The two nPCR-18Sa-positive samples were nPCR-ITS1-positive and nPCR-B1-negative. The nPCR-ITS1 nucleotide sequences from the two skuas, which were identical to each other, were revealed closely related to homologous sequences of Sarcocystis halieti, species found in seabirds of northern hemisphere. Larger fragments of genes encoding 18S and partial sequences of genes coding for cytochrome oxidase subunit 1 were also analyzed, corroborating ITS1 data. The haplotypes found in the skuas are unprecedent and closely related to species that use birds as the definitive host. Further studies need to be carried out to detect, identify and isolate this parasite to understand the epidemiology of the infection and its impact on the health of marine fauna.

Vertical transmission of Toxoplasma gondii in naturally infected ewes in the semiarid region of Brazil.

To evaluate transplacental transmission of Toxoplasma gondii in naturally infected ewes, blood samples were collected from 55 pregnant ewes and their offspring, before ingestion of colostrum. From 16 offspring of positive ewes and nine offspring from negative ewes, blood samples were obtained after 48 h and 7, 14, 21, 28, 35, 42, 49 and 56 days after birth. T. gondii antibodies were detected in serum samples using the indirect fluorescence antibody test (IFAT ≥ 64). Four of the 30 positive ewes (13.3 %) had offspring positive for T. gondii before ingesting colostrum (vertical transmission). The colostrum antibody titers decreased every week, and only 20 % (2/10) of the lambs in continued to present detectable antibody titers until day 56 after birth. Therefore, vertical transmission of T. gondii in lambs was indication of occur and is an important route for transferring and maintaining the agent in sheep herds in the Brazilian semiarid region.

Zoonotic parasites infecting free-living armadillos from Brazil.

Armadillos are specialist diggers and their burrows are used to find food, seek shelter and protect their pups. These burrows can also be shared with dozens of vertebrate and invertebrate species and; consequently, their parasites including the zoonotics. The aim of this study was to diagnose the presence of zoonotic parasites in four wild-caught armadillo species from two different Brazilian ecosystems, the Cerrado (Brazilian savanna) and the Pantanal (wetland). The investigated parasites and their correspondent diseases were: Toxoplasma gondii (toxoplasmosis), Trypanosoma cruzi (Chagas disease), Leishmania spp., (leishmaniasis), Paracoccidioides brasiliensis (Paracoccidioidomicosis) and Mycobacterium leprae (Hansen's disease). Forty-three free-living armadillos from Pantanal and seven road-killed armadillos from the Cerrado were sampled. Trypanosoma cruzi DTU TcIII were isolated from 2 out of 43 (4.65%) armadillos, including one of them also infected with Trypanosoma rangeli. Antibodies anti-T. gondii were detected in 13 out of 43 (30.2%) armadillos. All seven armadillos from Cerrado tested positive for P. brasiliensis DNA, in the lungs, spleen, liver fragments. Also, by molecular analysis, all 43 individuals were negative for M. leprae and Leishmania spp. Armadillos were infected by T. cruzi, T. rangeli, P. brasiliensis and presented seric antibodies to T. gondii, highlighting the importance of those armadillos could have in the epidemiology of zoonotic parasites.

Novel Ehrlichia sp. detected in Magellanic penguins (Sphenicus magellanicus) and in the seabird tick Ixodes uriae from Magdalena Island, southern Chile.

Ehrlichia spp. are obligatory intracellular microorganisms that infect hematopoietic, endothelial or blood cells of mammals. Ticks are the only vectors of these agents in nature. To date, the role of birds and their associated ticks as reservoirs of ehrlichiae remains almost unexplored. In this study, we performed a molecular screening for bacteria of Anaplasmataceae family in samples of spleen (n = 72) and lung (n = 17), recovered from 72 carcasses of Magellanic penguins (Spheniscus magellanicus) in Brazil and Chile. One apparently unengorged tick (Ixodes uriae) was also collected while wandering upon one of the carcasses and submitted to molecular analyses as well. Through conventional and nested PCR protocols three genes (16S rRNA, dsb and groEL) of a new Ehrlichia sp. were partially characterized upon organs of three penguins and in the tick coming from Magdalena Island (Chile). First matches after BLASTn comparisons showed that our sequences share 99.4% (16S rRNA), 94.6% (groEL) and 79.3% (dsb) of identity with "Candidatus Ehrlichia ornithorhynchi", Ehrlichia sp. NS101 and Ehrlichia canis CCZ, respectively. Matrixes of genetic distance including other representatives of the Ehrlichia genus point a 99.4%, 94.0%, and 80.0% of identity with 16S rRNA, groEL and dsb genes from Ehrlichia sp. It25, Ehrlichia sp. NS101, and Ehrlichia chaffeensis San Louis, respectively. A Bayesian phylogenetic analysis of Anaplasmataceae 16S rRNA gene places the detected Ehrlichia sp. into a group with Ehrlichia sp. BAT and Ehrlichia sp. Natal. Although depicting different topologies, Bayesian unrooted phylogenetic trees constructed for groEL and dsb genes position this Ehrlichia sp. into well-supported branches, which reinforces the finding of a new taxon. For the moment, any pathogenic effect of this new Ehrlichia sp. on penguins is still unknown. However, this fact becomes important to assess from a conservation point of view since populations of Magellanic penguins are currently threatened and in an ongoing decrease.

Clinical Toxoplasmosis in Dogs and Cats: An Update.

Toxoplasmosis is caused by the globally distributed protozoan parasite Toxoplasma gondii (phylum Apicomplexa); the disease can be clinically important for almost all homeothermic animals, including birds and humans. Toxoplasmosis course involves general clinical signs, such as fever, anorexia, or dyspnea, and more specific signs with neural, respiratory, cutaneous, or ocular involvement. Because of the wide range of clinical signs, the diagnosis in domestic and pet animals can be complicated. Hence, this review aims to provide a comprehensive analysis of some scarcely discussed aspects of toxoplasmosis, such as ocular and cutaneous manifestations, congenital infections, influence of T. gondii genotype on clinical toxoplasmosis, and recent findings regarding differential diagnosis. This review could be of special interest to clinicians and researchers.

Ticks, rickettsial and erlichial infection in small mammals from Atlantic forest remnants in northeastern Brazil.

We evaluated infection by Rickettsia spp. and Ehrlichia spp in small mammals and their ticks from two Atlantic forest conservation areas in the state of Rio Grande do Norte, northeastern Brazil. A total of 39 small mammals were captured during 2012-2013, encompassing 33 marsupials (29 Didelphis albiventris, four Monodelphis domestica), three Cricetidae rodents (two Necromys lasiurus, one Rattus rattus), one Caviomorpha rodent (Thrichomys apereoides) and two armadillos (Euphractus sexcinctus). The ticks Amblyomma auricularium, Ixodes loricatus, and Ornithodoros mimon were collected from D. albiventris, whereas only A. auricularium was collected from armadillos. Through immunofluorescence assay with Rickettsia spp. antigens, 6/28 (21%) D. albiventris and the single R. rattus specimen reacted to at least one rickettsial antigen, with highest seroprevalence and endpoint titers to Rickettsia amblyommatis. A total of 150 ticks (126 A. auricularium, nine I. loricatus, 15 O. mimon) was tested for rickettsial infection by PCR, which detected only R. amblyommatis in most of the A. auricularium ticks. Lung and spleen samples were collected from small mammals (two N. lasiurus, six D. albiventris, three M. domestica, one T. apereoides, one R. rattus) and were tested by PCR for Anaplasmataceae agents. The spleen from one D. albiventris contained a new ehrlichial agent, here named as Ehrlichia sp. strain Natal. Phylogenetic analysis inferred from the dsb gene of Ehrlichia spp. indicates that this novel agent is potentially a new species. Future studies should monitor the possible role of rickettsial and/or ehrlichial microorganisms as agents of emerging diseases in these degraded areas of Atlantic forest, just as has occurred with other agents in degraded areas of this biome in southeastern Brazil.

Fatal toxoplasmosis in a southern muriqui (Brachyteles arachnoides) from São Paulo state, Brazil: Pathological, immunohistochemical, and molecular characterization.

We report the pathological, immunohistochemical, and molecular features of fatal acute systemic toxoplasmosis in an adult, female, free-living southern muriqui (Brachyteles arachnoides) from São Paulo state, Brazil. PCR-RFLP genotyping analysis identified the #21 genotype of Toxoplasma gondii. This represents the first report of acute toxoplasmosis involving this genotype in humans and animals.

Detection of anti-Toxoplasma gondii antibodies in small wild mammals from preserved and non-preserved areas in the Caatinga biome, a semi-arid region of Northeast Brazil.

This study was conducted to determine the seroprevalence of anti-Toxoplasma gondii antibodies in 152 free-living small wild mammals from distinct regions in the Caatinga biome, a semi-arid region in the Northeast of Brazil: the National Park of Serra das Confusões (NPSC), which is a preserved area in the state of Piauí, and the municipalities of Petrolina and Lagoa Grande, two non-preserved areas in the state of Pernambuco. Using the modified agglutination test (MAT), we found that 5.3% (4/75) and 3.3% (2/60) of small wild mammals were positive for IgG anti-T. gondii antibodies in the NPSC and Petrolina, respectively. All mammals from Lagoa Grande (0/17) tested negative on the MAT. Indirect infection of T. gondii was determined by MAT in Galea spixii, Monodelphis domestica and Thrichomys laurentius (from NPSC) and in Didelphis albiventris (from Petrolina). Seropositive animals were observed in both preserved and non-preserved areas within the Caatinga biome. Low seroprevalences observed can be related to the extreme temperature and humidity in this particular biome.

Fatal toxoplasmosis in a southern muriqui (Brachyteles arachnoides) from São Paulo state, Brazil: Pathological, immunohistochemical, and molecular characterization

We report the pathological, immunohistochemical, and molecular features of fatal acute systemic toxoplasmosis in an adult, female, free­living southern muriqui (Brachyteles arachnoides) from São Paulo state, Brazil. PCR­RFLP genotyping analysis identified the #21 genotype of Toxoplasma gondii. This represents the first report of acute toxoplasmosis involving this genotype in humans and animals.

Relatamos as características patológicas, imuno-histoquímicas e moleculares da toxoplasmose sistêmica aguda fatal em um muriqui do sul adulto (feminino) de vida livre (Brachyteles arachnoides) do estado de São Paulo, Brasil. A análise de genotipagem por PCR ­ RFLP identificou o genótipo # 21 de Toxoplasma gondii. Isso representa o primeiro relato de toxoplasmose aguda envolvendo esse genótipo em humanos e animais

Pesquisa de anticorpos IgG para Neospora caninum e avaliação dos fatores de risco em ovinos do Estado de Sergipe / Neospora caninum IgG antibody research and evaluation of the risk factors associated to the infection in ovine, State of Sergipe, Brazil

O objetivo desse trabalho foi determinar a ocorrência de anticorpos IgG para Neospora caninum bem como avaliar os fatores de risco associados à infecção em rebanhos ovinos do estado de Sergipe, Brasil. Foram coletadas, nos anos de 2011 e 2012, 1200 amostras de sangue de ovinos oriundos de sessenta propriedades distribuídas em três mesorregiões do estado para pesquisa de anticorpos para N. caninum pela Reação de Imunofluorescência Indireta (RIFI) utilizando-se como ponto de corte de 50 e as amostras diluídas na base 2. Os dados de 34 variáveis estudadas foram obtidos a partir de questionários aplicados aos proprietários e analisados para se determinar a frequências absolutas e relativas e análise dos fatores de risco pelo teste Qui-quadrado de Pearson (p≤0,05). A ocorrência de ovinos soropositivos para N. caninum foi de 39,83% (478/1200). Em relação às mesorregiões a ocorrência de animais e propriedades positivas foi de, respectivamente, 55,88% (380/680) e 88,24% (30/34) na Leste; 21,42% (60/280) e 42,85% (6/14) no Agreste e 15,83% (38/240) e 41,67% (5/12) no Sertão. Os títulos de anticorpos variaram de 50, representando 96,02% (459/478) das amostras soropositivas, a 6400 (1/478). Dentre as variáveis significantes, na analise multivariada, que foram consideradas com fatores de risco para a infecção pelo N. caninum estavam propriedades localizadas na mesorregião Leste (p=0,000, OR=4,64, IC95%=3,36-6,41), presença de fonte de água parada e corrente (p=0,000 OR=2,03, IC95%=1,41-2,92), ausência de quarentena (p=0,000 OR=2,71, IC95%=2,08-3,53), não utilização de esterqueiras (p=0,000 OR=3,14, IC95%=2,45-4,02), criações com finalidade de subsistência (p=0,000 OR=4,99, IC95%=3,15-7,92), de reprodução (p=0,002, OR=1,74, IC95%=1,22-2,49), presença de cães (p=0,000 OR=2,74, IC95%=1,73-433) e circulação de animais silvestres nos rebanhos (p=0,000 OR=3,45, IC95%=2,44-4,87). Os resultados evidenciam a ocorrência de N. caninum em rebanhos ovinos sergipanos, demonstrando o manejo e a localização dos rebanhos no estado como importantes fatores de risco.(AU)

The aim of this study was to determine the occurrence of antibodies IgG against Neospora caninumand evaluate the risk factors associated with the infection in ovine herds, State of Sergipe, Brazil. Blood samples (n=1200) were collected from sheep raised in 60 sheep run located in the three mesoregions of the State of Sergipe, between 2011 e 2012. Antibodies were investigated by Indirect Immunofluorescence Antibody Test of which cutoff point was 50 and positive samples were diluted in base 2 until the last positive titer. Data from 15 variables was obtained from questionnaires given to farmers. Absolute and relative frequencies were determined and the risk factors were analyzed by Pearson's Qui-Square test (p≤0,05). The occurrences of serum reactive sheep were 39.83% (478/1200). The occurrences of positive sheep and sheep run were 55.88% (380/680) and 88.24% (30/34) in the Eastern region; 21.42% (60/280) e 42.85% (6/14) in dry region and 15.83% (38/240) e 41.67% (5/12) in the backwoods respectively. Antibody titers ranged from 50 (n=459), represented 96.02% (459/478) of seropositive samples to 6400 (1/478). Among the significant variables in the multivariate analysis were considered risk factors for infection with N. caninum were, sheep run located in Eastern region (p=0.000, OR=4.64, CI95%=3.36-6.41); standing and running water sources (p=0.000 OR=2.03, CI95%=1.41-2.92), absence of quarantine (p=0.000, OR=2.71, CI95%=2.08-3.53), absence of dunghill (p=0.000, OR=3.14, CI95%=2.45-4.02), presence of dogs (p=0.000, OR=2.74, CI95%=1.73-433), presence of wild animals (p=0.000, OR=3.45, CI95%=2.44-4.87) and subsistence (p=0.000, OR=4.99, CI95%=3.15-7.92) or reproduction (p=0.002, OR=1.74, CI95%=1.22-2.49) livestock were important risk factors. Our results highlight the occurrence ofN. caninumin the ovine herds from State of Sergipe. Management and location of sheep runs were important risk factors associated to the infection.(AU)

Novel Anaplasma and Ehrlichia organisms infecting the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals were sampled in the Amazon biome of Brazil. Animal tissues and blood were tested by polymerase chain reaction (PCR) assays targeting DNA of the bacterial family Anaplasmataceae (genera Anaplasma, Ehrlichia, Wolbachia) and the genus Borrelia. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. All birds and reptiles were negative by all PCR assays, as well as all mammals for the Borrelia PCR assay. Anaplasmataceae agents were searched by PCR assays targeting two different genes, the ribosomal 16S rRNA gene and the protein-coding dsb gene. Three dsb closely related haplotypes were generated from 3 white-lipped peccaries (Tayassu pecari). In a phylogenetic analysis inferred from dsb partial sequences, these haplotypes grouped with previously reported Ehrlichia haplotypes from jaguar (Panthera onca) and horse from Brazil, suggesting that they could all represent a single species, yet to be properly characterized. A unique dsb haplotype was generated from a sloth (Bradypus tridactylus), and could also represent a different Ehrlichia species. All these dsb haplotypes formed a clade sister to the Ehrlichia ruminantium clade. Three distinct 16S rRNA gene haplotypes were generated from a wild guinea pig (Cavia sp.), a woolly mouse opossum (Micoureus demerarae), and two from robust capuchin monkeys (Sapajus sp.). In a phylogenetic analysis inferred from 16S rRNA gene partial sequence, these haplotypes grouped within the Wolbachia clade, and are likely to represent Wolbachia organisms that were infecting invertebrate metazoarians (e.g., filarids) associated with the sampled mammals. Two deer (Mazama americana) samples yielded two distinct 16S rRNA gene sequences, one identical to several sequences of Anaplasma bovis, and an unique sequence that grouped in a clade with different Anaplasma species. Our results indicate that a variety of genetically distinct Anaplasmataceae organisms, including potentially new Ehrlichia species, circulate under natural conditions in the Amazonian wildlife.

Novel piroplasmid and Hepatozoon organisms infecting the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals were sampled in two areas within the Amazon biome of Brazil, in the states of Mato Grosso and Pará. Animal tissues and blood were molecularly tested for the presence of Piroplasmida (genera Babesia, Theileria, Cytauxzoon) or Hepatozoon DNA. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. The following Piroplasmida agents were detected: Cytauxzoon felis in one ocelot (Leopardus pardalis), Theileria cervi in two red brocket deer (Mazama americana), Theileria spp. in three nine-banded-armadillos (Dasypus novemcinctus), one agouti (Dasyprocta sp.), and four lowland pacas (Cuniculus paca), Babesia spp. in one common opossum (Didelphis marsupialis) and one white-lipped peccary (Tayassu pecari). The following Hepatozoon agents were detected: Hepatozoon sp. (possibly Hepatozoon caimani) in three spectacled caimans (Caiman crocodilus), Hepatozoon felis in an ocelot (Leopardus pardalis), and Hepatozoon spp. in one scorpion mud turtle (Kinosternon scorpioides) and one lowland paca (Cuniculus paca). Phylogenetic analyses inferred by the 18S rRNA gene partial sequences supported these results, highlighting at least five novel Piroplasmida agents, and two novel Hepatozoon agents. This study screened the presence of tick-borne protozoa in a number of wildlife species from the Amazon for the first time. Our results indicate that a variety of genetically distinct Piroplasmida and Hepatozoon organisms circulate under natural conditions in the Amazonian wildlife.

Seroprevalence of Toxoplasma gondii in seabirds from Abrolhos Archipelago, Brazil.

Toxoplasma gondii is a coccidian parasite that infects almost all warm-blooded animals, including birds. Abrolhos is an archipelago of five islands, located in the Atlantic Ocean, 56 nautical kilometers from the south coast of the state of Bahia, northeastern Brazil. Part of this archipelago is a National Marine Park, which is a conservation area protected by the Brazilian government. The objective of this study was to determine the occurrence of T. gondii antibodies in sera of seabird's species Sula spp. and Phaeton spp. from breeding colonies located in the Islands of Santa Bárbara and Redonda, Abrolhos's archipelago. Sera were tested by modified agglutination test, first screened at 1:5 dilution (cut-off point) and the positive samples were titrated at a two-fold serial dilution. Serum samples were obtained from 69 birds of four species: Sula dactylatra (23 birds), Sula leucogaster (19 birds), Phaeton aethereus (25 birds) and Phaeton lepturus (2 birds). Antibodies to T. gondii were found in 24 (34.8%) of 69 seabirds with titers that ranged from 5 to 640. Occurrence value in S. dactylatra was 34.8% (8/23), in S. leucogaster was 47.4% (9/19), in P. aethereus was 28% (7/25) and the 2 P. lepturus were negative. This is the first description of T. gondii antibodies in free ranging seabirds of the orders Suliformes and Phaethontiformes.

SEROPREVALENCE OF TOXOPLASMA GONDII IN CAPTIVE ANTILLEAN MANATEE (TRICHECHUS MANATUS MANATUS) IN BRAZIL.

Antillean manatees ( Trichechus manatus manatus) are aquatic mammals that inhabit marine waters from Central America to the northeastern region of Brazil, and they are an endangered species. Infection with Toxoplasma gondii through intake of water or food contaminated with oocysts has been reported among marine mammals. The present study aimed to evaluate the prevalence of antibodies to T. gondii in West Indian manatees living in captivity in northeastern Brazil. Serum samples from 55 West Indian manatees from three different captive groups were tested for T. gondii antibodies by means of the modified agglutination test using a cutoff of 1:25. The samples were screened at dilutions of 1:25, 1:50, and 1:500, and positive samples were end-titrated using twofold serial dilutions; antibodies were found in six Antillean manatees (10.9%) with titers of 1:50 in three, 1:500 in one, 1:3,200 in one, and 1:51,200 in one manatee. This study is the first report of T. gondii antibodies in captive Antillean manatees in Brazil.

Acute toxoplasmosis in pigs in Brazil caused by Toxoplasma gondii genotype Chinese 1.

This study reports the clinicopathological, immunohistochemical, and molecular findings from two cases of systemic toxoplasmosis in pigs showing apathy and dyspnea. In the post-mortem examination, severe diffuse necrotizing bronchointerstitial pneumonia with numerous intralesional tachyzoites of Toxoplasma gondii was observed. The lungs had not collapsed but were diffusely reddened, and the parenchyma showed friable whitish subpleural nodules with multifocal to coalescent distribution and diameters of 0.5-1.0 cm. The histopathological findings comprised mononuclear inflammation and multifocal areas of necrosis in alveolar septa (cases 1 and 2). In addition, esophagitis and ulcerations in the mucosa of the stomach and the small and large intestines were observed (case 1). Immunohistochemical analysis using anti-T. gondii antibodies on lung tissue in both cases revealed strong immunolabeling of free tachyzoites and tachyzoites in the cytoplasm of histiocytes and in cysts. Nested PCR targeting a 155-bp fragment of the B1 gene of T. gondii was positive for the DNA extracted from lung fragments from the two pigs. Genotyping of the samples by means of PCR-RFLP (10 markers) and by means of microsatellites (15 of them) revealed that these animals were infected with T. gondii that was molecularly characterized as the non-archetypal genotype Chinese 1. This presents worldwide circulation, but it had not previously been described in Brazil. The microsatellite analysis showed that the animals were infected with the same T. gondii isolate circulating in the environment.

SURVEY OF TOXOPLASMA GONDII ANTIBODIES IN MAGELLANIC PENGUINS (SPHENISCUS MAGELLANICUS FORSTER, 1781).

Magellanic penguins (Spheniscus magellanicus) breed on the Atlantic and Pacific coasts of the southernmost parts of South America and migrate northward as far as Peru and Brazil. Serum samples (n = 100) from Magellanic penguins from three zoos and two rehabilitation centers (RCs) in Brazil were assayed for the presence of antibodies to Toxoplasma gondii by means of the modified agglutination test (MAT, cut-off ≥ 20). The penguins were categorized as young (≤4 yr old) or adults (≥4 yr old) and sexed (male, female, or not identified), and data were analyzed using the chi-square test (P ≤ 0.05). Toxoplasma gondii antibodies were found in 28% of penguins: 25.8% males, 27.8% females, 30.3% unknown sex, 25.4% young, and 31.1% adults. Statistical analyses did not find any difference (P > 0.05) with respect to age, sex, or source of birds. This is the first report of T. gondii antibodies in S. magellanicus.

Local admixture of amplified and diversified secreted pathogenesis determinants shapes mosaic Toxoplasma gondii genomes.

Toxoplasma gondii is among the most prevalent parasites worldwide, infecting many wild and domestic animals and causing zoonotic infections in humans. T. gondii differs substantially in its broad distribution from closely related parasites that typically have narrow, specialized host ranges. To elucidate the genetic basis for these differences, we compared the genomes of 62 globally distributed T. gondii isolates to several closely related coccidian parasites. Our findings reveal that tandem amplification and diversification of secretory pathogenesis determinants is the primary feature that distinguishes the closely related genomes of these biologically diverse parasites. We further show that the unusual population structure of T. gondii is characterized by clade-specific inheritance of large conserved haploblocks that are significantly enriched in tandemly clustered secretory pathogenesis determinants. The shared inheritance of these conserved haploblocks, which show a different ancestry than the genome as a whole, may thus influence transmission, host range and pathogenicity.

The ROP18 and ROP5 gene allele types are highly predictive of virulence in mice across globally distributed strains of Toxoplasma gondii.

The protozoan parasite Toxoplasma gondii is one of the most successful known eukaryotic pathogens on Earth. Virulence of T. gondii strains varies greatly in mice, and mounting evidence suggests that such variations may be relevant to the manifestation of human toxoplasmosis. Polymorphic rhoptry-secreted kinases and pseudokinases (ROP) have been demonstrated to account for murine virulence among the archetypal clonal parasite lineages that dominate the populations of North America and Europe. However, the distribution of virulence gene alleles in natural populations and the broad influence of these allele combinations on T. gondii virulence have not been examined in depth. In the present study, we performed PCR-RFLP genotyping analysis on a diverse array of globally distributed T. gondii strains at four ROP gene loci including ROP18, ROP5, ROP16 and ROP17 that were previously implicated in influencing T. gondii virulence and pathogenesis. We demonstrated through correlation with published virulence data that the combination of ROP18 and ROP5 allele types is highly predictive of T. gondii virulence across a broad range of global T. gondii isolates. These findings indicate that the importance of ROP18 and ROP5 in determining strain virulence is not limited to the North American/European archetypal lineages most commonly used in molecular studies, but also appears to apply to diverse isolates from South/central America and Asia. Restriction fragment length polymorphism analysis of these loci may thus serve as a valuable tool in determining the potential virulence of uncharacterized T. gondii strains in future studies.