Exploring interactions between parasites and their hosts in the Pantanal floodplain using an ecological network approach.

The study of host-parasite interactions is essential to understand the role of each host species in the parasitic transmission cycles in a given community. The use of ecological network highlights the patterns of interactions between hosts and parasites, allowing us to evaluate the underlying structural features and epidemiological roles of different species within this context. Through network analysis, we aimed to understand the epidemiological roles of mammalian hosts species (n = 67) and their parasites (n = 257) in the Pantanal biome. Our analysis revealed a modular pattern within the network, characterized by 14 distinct modules, as well as nestedness patterns within these modules. Some key nodes, such as the multi-host parasites Trypanosoma cruzi and T. evansi, connect different modules and species. These central nodes showed us that various hosts species, including those with high local abundances, contribute to parasite maintenance. Ectoparasites, such as ticks and fleas, exhibit connections that reflect their roles as vectors of certain parasites. Overall, our findings contribute to a comprehensive understanding of the structure of host-parasite interactions in the Pantanal ecosystem, highlighting the importance of network analysis as a tool to identifying the main transmission routes and maintenance of parasites pathways. Such insights are valuable for parasitic disease control and prevention strategies and shed light on the broader complexities of ecological communities.

Contribution to the knowledge of Neotrichodectes (Nasuicola) pallidus (Piaget, 1880) (Phthiraptera: Trichodectidae).

The species in the genus Neotrichodectes (Phthiraptera: Ischnocera) infest carnivores. Neotrichodectes (Nasuicola) pallidus (Piaget, 1880), which has been primarily found parasitizing Procyonidae mammals, has been recorded in ring-tailed coatis (Nasua nasua) in the Brazilian states of Minas Gerais, Pernambuco, Santa Catarina, Rio Grande do Sul and Pernambuco. We report a new record of N. pallidus in coatis in the state of Mato Grosso do Sul, central-western Brazil, using morphological (Light and Scanning Electronic Microscopy) and molecular approaches (PCR, sequencing and phylogenetic analysis). Coatis were sampled in two peri-urban areas of Campo Grande city, Mato Grosso do Sul state, Brazil, between March 2018 and March 2019, as well as in November 2021. Lice were collected and examined under light and Scanning Electron Microscopy. DNA was also extracted from nymphs and adults and submitted to PCR assays based on the 18S rRNA and cox-1 genes for molecular characterization. One hundred and one coatis were sampled from 2018 to 2019 and 20 coatis in 2021 [when the intensity of infestation (II) was not accessed]. Twenty-six coatis (26/101-25.7%) were infested with at least one louse, with a total of 59 lice collected in 2018-2019. The II ranged from one to seven lice (mean 2.2 ± SD 1.7). The louse species was confirmed based on the following morphological characteristics: female gonapophyses rounded with the setae along anterior region but not in the medial margin; the male genitalia with a parameral arch not extending beyond the endometrial plate. The same ornamentation was observed on the abdomen of the females, males, and nymphs. The nymphs and the eggs were described in detail for the first time. The obtained 18S rRNA and cox1 sequences from N. pallidus clustered in a clade with other sequences of Ischnocera species. In the present study, a new record of the louse N. pallidus in central-western Brazil was provided, along with new insights into the morphological features of this species, with the first morphology contribution of nymphal and eggs stages.

Molecular Survey of Hemotropic Mycoplasma spp. and Bartonella spp. in Coatis (Nasua nasua) from Central-Western Brazil.

Even though previous works showed molecular evidence of hemotropic Mycoplasma spp. (hemoplasmas) in ring-tailed coatis (Nasua nasua) from Brazil, Bartonella sp. has not been reported in these mammals so far. The present study aimed to detect the above-mentioned agents in coatis' blood and associated ectoparasites, assessing the association between these infections and red blood parameters. Between March 2018 and January 2019, coati (n = 97) blood samples, Amblyomma sp. ticks (2242 individual ticks, resulting in 265 pools), and Neotrichodectes pallidus louse (n = 59) were collected in forested urban areas from midwestern Brazil. DNA extracted from coatis' blood, and ectoparasite samples were submitted to quantitative PCR (qPCR) (16S rRNA) and conventional PCR (cPCR) (16S rRNA and 23S rRNA) for hemoplasmas and qPCR (nuoG gene) and culturing (only blood) for Bartonella spp. Two different hemoplasma genotypes were detected in blood samples: 71% coatis positive for myc1 and 17% positive for myc2. While 10% of ticks were positive for hemoplasmas (myc1), no louse was positive. The estimated bacterial load of hemoplasmas showed no association with anemia indicators. All coatis were negative for Bartonella sp. in qPCR assay and culturing, albeit two Amblyomma sp. larvae pools, and 2 A. dubitatum nymph pools were positive in the qPCR. The present work showed a high occurrence of hemoplasmas, with two distinct hemoplasma genotypes, in coatis from forested urban areas in midwestern Brazil.

Corrigendum: Leishmania infantum infecting the carnivore Nasua nasua from urban forest fragments in an endemic area of visceral leishmaniasis in Brazilian Midwest.

[This corrects the article DOI: 10.3389/fcimb.2022.1050339.].

Contribution to the knowledge of Neotrichodectes (Nasuicola) pallidus (Piaget, 1880) (Phthiraptera: Trichodectidae)

The species in the genus Neotrichodectes (Phthiraptera: Ischnocera) infest carnivores. Neotrichodectes (Nasuicola) pallidus (Piaget, 1880), which has been primarily found parasitizing Procyonidae mammals, has been recorded in ring-tailed coatis (Nasua nasua) in the Brazilian states of Minas Gerais, Pernambuco, Santa Catarina, Rio Grande do Sul and Pernambuco. We report a new record of N. pallidus in coatis in the state of Mato Grosso do Sul, central-western Brazil, using morphological (Light and Scanning Electronic Microscopy) and molecular approaches (PCR, sequencing and phylogenetic analysis). Coatis were sampled in two peri-urban areas of Campo Grande city, Mato Grosso do Sul state, Brazil, between March 2018 and March 2019, as well as in November 2021. Lice were collected and examined under light and Scanning Electron Microscopy. DNA was also extracted from nymphs and adults and submitted to PCR assays based on the 18S rRNA and cox-1 genes for molecular characterization. One hundred and one coatis were sampled from 2018 to 2019 and 20 coatis in 2021 [when the intensity of infestation (II) was not accessed]. Twenty-six coatis (26/101–25.7%) were infested with at least one louse, with a total of 59 lice collected in 2018–2019. The II ranged from one to seven lice (mean 2.2 ± SD 1.7). The louse species was confirmed based on the following morphological characteristics: female gonapophyses rounded with the setae along anterior region but not in the medial margin; the male genitalia with a parameral arch not extending beyond the endometrial plate. The same ornamentation was observed on the abdomen of the females, males, and nymphs. The nymphs and the eggs were described in detail for the first time. The obtained 18S rRNA and cox1 sequences from N. pallidus clustered in a clade with other sequences of Ischnocera species. In the present study, a new record of the louse N. pallidus in central-western Brazil was provided, along with new insights into the morphological features of this species, with the first morphology contribution of nymphal and eggs stages.

Brucellosis in the Brazilian Pantanal wetland: threat to animal production and wildlife conservation.

This review was performed to gather knowledge about brucellosis in livestock and wildlife in the Brazilian Pantanal, a biome with a huge biodiversity and extensive livestock production. Following the preferred reporting items for narrative review guidelines and using the terms "Brucella" and "Pantanal," we explored the PubMed, SciELO, Jstor, Science Direct, and Scholar Google databases. Information on host species, diagnostic test, number of positive animals, and positivity rates were acquired. Articles dating from 1998 to 2022 registered 14 studies including cattle, dogs, and the following wildlife species: Ozotoceros bezoarticus, Sus scrofa, Tayassu peccari, Nasua nasua, Cerdocyon thous, Panthera onca, Dasypus novemcintus, Cabassous unicinctus, Euphractus sexcinctus, Priodontes maximus, Myrmecophaga tridactyla and Hydrochoerus hydrochaeris. Brucella occurrence in cattle was demonstrated through the serological confirmatory test 2-mercaptoetanol. Molecular diagnosis detected Brucella abortus in dogs, smooth Brucella in O. beoarticus, and Brucella spp. in T. peccari. Cattle may have a pivotal importance in maintenance and spreading of Brucella spp. due to their high population density, environmental contamination from abortion of infected cows, and eventual excretion of B. abortus S19 strain from vaccinated heifers. The occurrence of Brucella spp. in O. bezoarticus and T. peccari represent a risk for wildlife conservation. These data indicate that Brucella spp. are enzootic in the Pantanal wetland, sustained by a reservoir system including domestic and wild mammals. Due to marked seasonality and large populations of wildlife species sharing same environments with livestock, brucellosis acquires great complexity in Pantanal and, therefore, must be analyzed considering both animal production and conservation.

Leishmania infantum infecting the carnivore Nasua nasua from urban forest fragments in an endemic area of visceral leishmaniasis in Brazilian Midwest.

Introduction: The aim of the present study was to investigate the occurrence of Leishmania infantum in South American coatis inhabiting two forest fragments in Campo Grande, Mato Grosso do Sul, Midwest region of Brazil, an endemic area of human and canine visceral leishmaniasis (VL). Material and methods: A total of 110 South American coatis were sampled in the conservation unit "Parque Estadual do Prosa" (PEP) and in the residential area "Vila da Base Aérea" (VBA) from March 2018 to April 2019. As a longitudinal study that include up to six recaptures of the same individual, a total of 190 capture events were obtained. Blood, bone marrow and skin samples were obtained for parasitological (axenic culture), serological (Enzyme Linked Immunosorbent Assay - ELISA and Dual-path Platform immunoassay - DPP® CVL) and molecular diagnostic assays (targeting kDNA for Leishmania spp. and L. infantum; and HSP70 followed by sequence analysis). Results: Seropositivity for L. infantum was found in 33 individuals, six in PEP and 27 in VBA. Furthermore, L. infantum was detected by molecular analysis in 16 individuals, seven from PEP and nine from VBA. We also isolated L. infantum from bone marrow of one individual and detected a single positive skin sample in molecular assay from other individual, both from VBA. Discussion: An overall infection rate of 36.4% (40/110) was observed, significantly higher in the VBA (49.1%) than in the PEP (21.6%), probably because VBA presents: (i) a large number of resident dogs and chickens that would be attracting sandflies; (ii) a denser population of this wild mammal species; and (iii) physical barriers and a lack of functional connectivity in the surroundings, preventing these animals to disperse out. We conclude that South American coati populations living in urban forest fragments of Campo Grande are affected by the epidemiological scenario of VL, known to involve dogs, vectors and humans. We highlight the importance of investigate the parasitism by L. infantum in this and other potential L. infantum reservoirs that inhabit urbanized regions endemic to VL.

Trypanosomatid species in Didelphis albiventris from urban forest fragments.

Urbanization results in loss of natural habitats and, consequently, reduction of richness and abundance of specialist to the detriment of generalist species. We hypothesized that a greater richness of trypanosomatid in Didelphis albiventris would be found in fragments of urban forests in Campo Grande, Mato Grosso do Sul, Brazil, that presented a larger richness of small mammals. We used parasitological, molecular, and serological methods to detect Trypanosoma spp. infection in D. albiventris (n = 43) from forest fragments. PCR was performed with primers specific for 18S rDNA, 24Sα rDNA, mini-chromosome satellites, and mini-exon genes. IFAT was used to detect anti-Trypanosoma cruzi IgG. All hemoculture was negative. We detected trypanosomatid DNA in blood of 35% of opossum. Two opossums were seropositive for T. cruzi. The trypanosomatid species number infecting D. albiventris was higher in the areas with greater abundance, rather than richness of small mammals. We found D. albiventris parasitized by T. cruzi in single and co-infections with Leishmania spp., recently described molecular operational taxonomic unit (MOTU) named DID, and Trypanosoma lainsoni. We concluded that (i) trypanosome richness may be determined by small mammal abundance, (ii) D. albiventris confirmed to be bio-accumulators of trypanosomatids, and (iii) T. lainsoni demonstrated a higher host range than described up to the present.

Low occurrence of Bartonella in synanthropic mammals and associated ectoparasites in peri-urban areas from Central-Western and Southern Brazil.

Worldwide, Bartonella species are known to infect a wide range of mammalian and arthropod hosts, including humans. The current study aimed to investigate the prevalence of Bartonella spp. in synanthropic mammals captured in peri-urban areas from Central-Western and Southern Brazil and their ectoparasites. For this aim, 160 mammals belonging to four species, and 218 associated arthropods were sampled. DNA was extracted and subjected to different Bartonella screening assays. Additionally, blood samples from 48 small rodents were submitted to liquid BAPGM culture followed by qPCR assay and solid culture. Two out of 55 Rattus captured in Santa Catarina state were PCR-positive for Bartonella when targeting the nuoG, 16S, and ITS loci. Sequences showed high homology with Bartonella coopersplainsensis. Conversely, all 48 small rodents, 14 capybaras and 43 opossum DNA samples from animals trapped in Mato Grosso do Sul were Bartonella negative in the HRM real time PCR assays targeting the ITS locus and gltA gene. Additionally, all mammal-associated ectoparasites showed negativity results based on HRM real time PCR assays. The present study showed, for the first time, the occurrence of B. coopersplainsensis in Brazil, shedding some light on the distribution of rats-related Bartonella in South America. In addition, the majority of rodents and marsupials were negative for Bartonella spp. Since B. coopersplainsensis reservoirs - Rattus spp. - are widely dispersed around the globe, their zoonotic potential should be further investigated.

Genetic diversity and lack of molecular evidence for hemoplasma cross-species transmission between wild and synanthropic mammals from Central-Western Brazil.

Globally, hemotropic mycoplasmas (hemoplasmas) comprise an emerging or remerging bacteria group that attaches to red blood cells of several mammal's species and in some cases, causing hemolytic anemia. Herein, we assessed the occurrence, genetic diversity, the factors coupled to mammals infection, and the phylogeographic distribution of hemoplasmas in sylvatic and synanthropic mammals and their associated ectoparasites from Brazil. We collected spleen and/or blood samples from synanthropic rodents (Rattus rattus [N = 39] and Mus musculus [N = 9]), sylvatic rodents (Hydrochoerus hydrochaeris [N = 14]) and opossums (Didelphis albiventris [N = 43]). In addition, ticks (Amblyomma spp. [N = 270] and lice (Polyplax spinulosa [N = 6]) specimens were also sampled. Using a PCR targeting the 16S rRNA region, out of 48 small rodents, 14 capybaras and 43 opossums DNA samples, hemoplasma DNA was found in 25%, 50%, and 32.5% animals, respectively. Besides, we reported hemoplasma DNA in Amblyomma sp. (22.2% [2/9]) and lice (100% [2/2]) pools samples from rats, and one female A. sculptum DNA sample (3% [1/33]) obtained from a capybara. Additionally, and in agreement with ML analysis, the network analyses showed a clear phylogenetic separation among the hemoplasmas genotypes found in the different host species sampled, thus, suggesting the absence of cross-species hemoplasmas transmission between the mammals trapped. Finally, using the NTC network analysis, we reported the same 16S rRNA Mycoplasma genotype circulating in Rattus sampled in Brazil, Hungary, and Japan.

The reservoir system for Trypanosoma (Kinetoplastida, Trypanosomatidae) species in large neotropical wetland.

Distinct species of Trypanosoma have been documented sharing the same hosts in different environments in intricate transmission networks. Knowing this, this study investigated the role of different hosts in the transmission cycles of Trypanosoma species in the Pantanal biome. The mammals were sampled from November 2015 to October 2016. We sampled a total of 272 wild mammals from 27 species belonging to six orders and 15 families, and three species of triatomines (n = 7). We found high parasitemias by Hemoculture test for Trypanosoma cruzi (TcI), Trypanosoma rangeli, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, and high parasitemias by Microhematocrit Centrifuge Technique for Trypanosoma evansi. The carnivore Nasua nasua is a key host in the transmission cycles since it displayed high parasitemias for T. cruzi, T. evansi and T. rangeli. This is the first report of high parasitemias in Tamandua tetradactyla and cryptic infection in Dasypus novemcinctus by T. cruzi; cryptic infection by T. evansi in Eira barbara, Euphractus sexcinctus and Dasyprocta azarae. The collection of Panstrongylus geniculatus increased the geographic distribution of this vector species in the South America. Our results indicate that Trypanosoma species circulate in a complex reservoir system including different host species with different infective competences.

Genetic diversity of Babesia bovis in beef cattle in a large wetland in Brazil.

Babesia bovis is the etiological agent of bovine babesiosis, a disease transmitted by Rhipicephalus microplus, which affects cattle herds in tropical and subtropical regions of the world, causing significant economic losses due to decreasing meat and milk yield. This study used molecular techniques to determine the occurrence and genetic diversity of B. bovis, based on the genes encoding the spherical body protein (sbp-2) and the merozoite surface antigens (MSAs) genes, in a herd of 400 Nellore (Bos indicus) sampled from beef cattle farms in the Pantanal region, state of Mato Grosso do Sul, Midwestern Brazil. The results of the nested PCR assays based on the sbp-2 gene indicated that 18 (4.5%) calves were positive for B. bovis; out of them, while 77.7% (14/18) were positive for the B. bovis msa-2b fragment, 66.6% (12/18) were positive for the msa-2c fragment. The phylogenetic analysis based on the maximum likelihood method using 14 sequences from msa-2b clones and 13 sequences from msa-2c clones indicated that the sequences detected in this study are clearly distributed in different cladograms. These findings corroborated the diversity analysis of the same sequences, which revealed the presence of 14 and 11 haplotypes of the msa-2b and msa-2c genes, respectively. Furthermore, the entropy analyses of amino acid sequences revealed 78 and 44 high entropy peaks with values ranging from 0.25 to 1.53 and from 0.27 to 1.09 for MSA-2B and MSA-2C, respectively. Therefore, the results indicate a low molecular occurrence of B. bovis in beef cattle sampled in the Brazilian Pantanal. Despite this, a high degree of genetic diversity was found in the analyzed B. bovis population, with possibly different haplotypes coexisting in the same animal and/or in the same studied herd.

Genetic diversity of Anaplasma marginale in beef cattle in the Brazilian Pantanal.

There are few studies on the genetic diversity of Anaplasma marginale in Brazilian cattle herds, especially about beef cattle. The objective of this study was to evaluate the genetic diversity of A. marginale, based on the msp1α gene in Bos taurus indicus sampled from the Brazilian Pantanal. Aliquots of blood with and without EDTA were taken from 400 cattle (200 cows and 200 calves) across five extensive farms. The samples were submitted to the indirect immunoenzymatic assay (iELISA), quantitative real-time PCR (qPCR) for the msp1ß gene and to the semi-nested (sn) PCR for the msp1α gene. Positive samples were sequenced by the Sanger method and subjected to diversity analysis using the RepeatAnalyser software. The percentage of positive animals by iELISA, qPCR and (sn) PCR was 72.2% (289/400), 56.7% (227/400) and 23% (52/227), respectively. Cows (154/200) showed to be significantly more seropositive than calves (135/200). In qPCR, the number of calves and average quantification value (138/200; 1.3 × 106) A. marginale msp1α copies per µL proved to be higher when compared to that found for the cows (89/200; 3.9 × 104). The microsatellite analysis of the 26 sequences obtained from the msp1α gene revealed the presence of E (77%), C (15.4%) and B (7.7%) genotypes. Fourteen A. marginale strains were identified in the studied region, with eight that have never before been described in the literature (τ-10-13-13-18; τ-27-18; EV8-EV8-17; α-ß-ß-ß-100; EV7-11-10-15; τ-11-11-27-18; τ-11-10-15; τ-27-13-18). Beef cattle are highly exposed to A. marginale in the Brazilian Pantanal. Moreover, a high genetic diversity of A. marginale, with eight new strains, was found in the studied region. While cows may act as chronic carriers, perpetuating the pathogen within the herd, male beef calves sold to other regions may disperse these strains.

Detection of Brucella spp. in dogs at Pantanal wetlands.

Canine brucellosis is an infectious disease that produces reproductive disease in both males and females. Although Brucella canis is more common, the infection by Brucella abortus is more frequent in dogs sharing habitats with livestock and wild animals. We decided to investigate the role of dogs in the maintenance of Brucella spp. in the Pantanal wetland. Serum and whole blood samples were collected from 167 dogs. To detect antibodies against B. abortus and B. canis, buffered acidified plate antigen (BAPA) and agar gel immunodiffusion (AGID) tests were performed. To detect Brucella spp., B. abortus and B. canis DNA, PCR was performed using the bcsp31, BruAb2_0168, and BR00953 genes, respectively. To confirm the PCR results, three bcsp31 PCR products were sequenced and compared with sequences deposited in GenBank. The seropositivity rates of 7.8% and 9% were observed for the AGID and BAPA tests, respectively. Positivity rates of 45.5% and 10.8% were observed when testing bcsp31 and BruAb2_0168, respectively, while there was no positivity for BR00953. The sequenced products had 110 base pairs that aligned with 100% identity to B. abortus, B. canis, and B. suis. Considering our results, dogs may be acting as maintenance hosts of Brucella spp. in the Pantanal region.

Maintenance of Trypanosoma cruzi, T. evansi and Leishmania spp. by domestic dogs and wild mammals in a rural settlement in Brazil-Bolivian border.

Domestic dogs are considered reservoirs hosts for several vector-borne parasites. This study aimed to evaluate the role of domestic dogs as hosts for Trypanosoma cruzi, Trypanosoma evansi and Leishmania spp. in single and co-infections in the Urucum settlement, near the Brazil-Bolivian border. Additionally, we evaluated the involvement of wild mammals' in the maintenance of these parasites in the study area. Blood samples of dogs (n = 62) and six species of wild mammals (n = 36) were collected in July and August of 2015. The infections were assessed using parasitological, serological and molecular tests. Clinical examination of dogs was performed and their feeding habits were noted. Overall, 87% (54/62) of sampled dogs were positive for at least one trypanosomatid species, in single (n = 9) and co-infections (n = 45). We found that 76% of dogs were positive for T. cruzi, four of them displayed high parasitemias demonstrated by hemoculture, including one strain types TcI, two TcIII and one TcIII/TcV. Around 73% (45/62) of dogs were positive to T. evansi, three with high parasitemias as seen by positive microhematocrit centrifuge technique. Of dogs sampled, 50% (31/62) were positive for Leishmania spp. by PCR or serology. We found a positive influence of (i) T. evansi on mucous pallor, (ii) co-infection by T. cruzi and Leishmania with onychogryphosis, and (iii) all parasites to skin lesions of sampled dogs. Finally, feeding on wild mammals had a positive influence in the Leishmania spp. infection in dogs. We found that 28% (5/18) coati Nasua nasua was co-infected for all three trypanosamatids, demonstrating that it might play a key role in maintenance of these parasites. Our results showed the importance of Urucum region as a hotspot for T. cruzi, T. evansi and Leishmania spp. and demonstrated that dogs can be considered as incidental hosts.

Correction: Outcomes of Trypanosoma cruzi and Trypanosoma evansi infections on health of Southern coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis) in the Brazilian Pantanal.

[This corrects the article DOI: 10.1371/journal.pone.0201357.].

Outcomes of Trypanosoma cruzi and Trypanosoma evansi infections on health of Southern coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis) in the Brazilian Pantanal.

The occurrence of Trypanosoma spp. in wild carnivore populations has been intensively investigated during the last decades. However, the impact of these parasites on the health of free-living infected animals has been largely neglected. The Pantanal biome is the world's largest seasonal wetland, harboring a great diversity of species and habitats. This includes 174 species of mammals, of which 20 belong to the order Carnivora. The present study aimed to investigate the effect of Trypanosoma evansi and Trypanosoma cruzi infections and coinfections on the health of the most abundant carnivores in the Pantanal: coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis). We captured 39 coatis, 48 crab-eating foxes, and 19 ocelots. Diagnostic tests showed T. cruzi infection in 7 crab-eating foxes and 5 coatis. Additionally, 7 crab-eating foxes, 10 coatis, and 12 ocelots were positive for T. evansi. We observed coinfections in 9 crab-eating foxes, 8 coatis, and 2 ocelots. This is the first report of T. evansi and T. cruzi infection on the health of free-living ocelots and crab-eating foxes. We showed that single T. evansi or T. cruzi infection, as well as coinfection, caused some degree of anemia in all animals, as well as an indirect negative effect on body condition in coatis and crab-eating foxes via anemia indicators and immune investment, respectively. Furthermore, the vigorous immune investment observed in sampled coatis, crab-eating foxes and ocelots infected by T. evansi, T. cruzi and coinfected can be highly harmful to their health. Overall, our results indicate that single and combined infection with T. evansi and T. cruzi represent a severe risk to the health of wild carnivores in the Pantanal region.

Assessment of a quantitative 5' nuclease real-time polymerase chain reaction using the nicotinamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) for Bartonella species in domiciled and stray cats in Brazil.

OBJECTIVES: The objective of this study was to develop a quantitative 5' nuclease real-time polymerase chain reaction (PCR) assay to diagnose infections caused by Bartonella species. METHODS: Between January and April 2013 whole blood samples were collected by convenience from 151 cats (86 domiciled and 65 stray cats). The feline blood samples were subjected to a novel quantitative 5' nuclease real-time PCR (qPCR) for Bartonella species targeting the nictonamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) and conventional PCR assays targeting intergenic transcribed spacer, ribC, gltA, pap31 and rpoB, followed by sequencing and basic local alignment search tool analysis. RESULTS: The qPCR assay detected as few as 10 copies of plasmid per reaction. Forty-six (54.4% domiciled and 45.6% stray cats) of 151 sampled cats showed positive results in nuoG qPCR for Bartonella species. The absolute quantification of nuoG Bartonella DNA in sampled cats ranged from 1.1 × 10(4) to 1.3 × 10(4). Eighteen (39.1%) of 46 positive samples in the qPCR were also positive in conventional PCR assays. The sequencing confirmed that Bartonella henselae and Bartonella clarridgeiae circulate in cats in midwestern Brazil. CONCLUSIONS AND RELEVANCE: The present work provides details of a novel qPCR assay to diagnose infections caused by Bartonella species.

Tick-borne agents in domesticated and stray cats from the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil.

Anaplasmataceae agents, piroplasmids and Hepatozoon spp. have emerged as important pathogens among domestic and wild felines. The present work aimed to detect the presence of species belonging to the Anaplasmataceae family, piroplasmas and Hepatozoon spp. DNA in blood samples of domesticated and stray cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Between January and April 2013, whole blood samples were collected from 151 cats (54 males, 95 females and two without gender registration) in the city of Campo Grande, state of Mato Grosso do Sul, Brazil. DNA extracted from cat blood samples was submitted to conventional PCR assays for Theileria/Babesia/Cytauxzoon spp. (18S rRNA, ITS-1), Ehrlichia spp. (16S rRNA, dsb, groESL), Anaplasma spp. (16S rRNA, groESL) and Hepatozoon spp. (18S rRNA) followed by phylogenetic reconstructions. Out of 151 sampled cats, 13 (8.5%) were positive for Ehrlichia spp. closely related to Ehrlichia canis, 1 (0.66%) for Hepatozoon spp. closely related to Hepatozoon americanum and Hepatozoon spp. isolate from a wild felid, 1 (0.66%) for Cytauxzoon sp. closely related do Cytauxzoon felis, and 18 (11.9%) for Babesia/Theileria (one sequence was closely related to Babesia bigemina, eight for Babesia vogeli, five to Theileria spp. from ruminants [Theileria ovis, Theileria lestoquardi] and four to Theileria sp. recently detected in a cat). The present study showed that Ehrlichia spp., piroplasmids (B. vogeli, Theileria spp. and Cytauxzoon spp.) and, more rarely, Hepatozoon spp. circulate among stray and domesticated cats in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil.