Co-infection by multiple vector-borne agents in wild ring-tailed coatis (Nasua nasua) from Iguaçu National Park, southern Brazil.

The present study aimed to detect molecularly the presence of co-infections by vector-borne agents (VBA) in ring-tailed coatis' (Nasua nasua) blood samples from Iguaçu National Park (INP), southern Brazil, and assess the phylogenetic positioning of the detected agents. DNA blood samples were submitted to molecular screening and characterization for Anaplasmataceae agents, Piroplasmids, Hepatozoon sp., hemotropic mycoplasmas, and Bartonella spp. In total, 42 (85.7%) coatis were positive for hemotropic Mycoplasma sp., 12 (24.5%) for Bartonella machadoae, 7 (14.3%) for Anaplasma sp. closely related to 'Candidatus Anaplasma brasiliensis', and 3 (6%) for Hepatozoon procyonis. The most prevalent co-infections observed was from bacterial VBA: while 18.3% were co-infected by hemotropic Mycoplasma sp. and Bartonella sp., 12.2% were co-infected by Anaplasma sp. and hemotropic Mycoplasma sp. Only two animals (4%) presented co-infections by three VBA (Bartonella sp., Anaplasma sp. and hemotropic Mycoplasma sp.). The coati is a wild carnivore found in INP, mainly in areas visited by tourists. These animals are frequently seen searching for food in garbage dumps or in tourists' belongings. The present study expands the host specificity range of B. machadoae, which has been isolated only from rodents until the present moment. Since the zoonotic potential and transmission routes of the detected VBA are not yet known, surveillance in this area is much needed.

Isolation of microorganisms from the feces of ring-tailed coati related to the production of "misha coffee" in the central forest of Peru and evaluation of some features of technological importance.

A diversity of yeasts and lactic bacteria were isolated from the feces of ring-tailed coaties bred in captivity and related to the production of "misha coffee". Isolation of yeasts was carried out using oxytetracycline-glucose-yeast extract agar containing 100 mg/L oxytetracycline and, lactic bacteria using de Man-Rogosa and Sharpe agar containing 20 mg/L of vancomicin. Then, isolates were biochemically analysed using API strips (ID 32C for yeasts and 50CHL for lactic bacteria) followed by 16S and 26S rRNA gene sequencing. Among the yeasts, Debaryomyces hansenii, Pichia kluyveri, Pichia kudriavzevii, and Candida sorboxilosa were the most frequent, whereas Weissella cibaria, Weissella paramesenteroides, Enterococcus thailandicus and Enterococcus faecalis were the most important lactic bacteria. Cultivation of the isolated yeasts under agitated conditions, showed that Pichia kluyveri LBFT.Lev3 (0.15 ± 0.01 h-1) and Pichia kudriavzevii LBTF.Lev7 (0.14 ± 0.01 h-1) had higher specific growth rates than Debaryomyces hansenii LBFT.Lev9 (0.09 ± 0.01 h-1), whereas cultivation of lactic bacteria under static fashion showed that Weisella paramesenteroides LBTF.Bal2 (0.16 ± 0.01 h-1) and Weisella cibaria LBTF.Bal3 (0.18 ± 0.01 h-1) had better growth than Enterococcus thailandicus LBTF.Bal1 (0.1 ± 0.015 h-1) and Enterococcus faecalis LBTF.Bal7 (0.14 ± 0.01 h-1). Additionally, evaluation of pectinolytic activity revealed that Pichia kudriavzevii LBTF.Lev7 and Debaryomyces hansenii LBFT.Lev9 were able to use pectin as carbon source for their growth. On the other hand, W. cibaria LBTF.Bal3, E. thailandicus LBTF.Bal1 and W. paramesenteroides LBTF.Bal2 showed inhibitory activity against S. mutans ATCC 35668, B. subtilis subsp. spizizenii ATCC 6633 and Staph. epidermidis ATCC 14990. Results of this study are useful for the search of potential application of the isolated yeasts and lactic bacteria in coffee and other food fermentations.

Hand Cellulitis and Abscess From a Kinkajou Bite: a Case Report and Review of Kinkajou Bites in Humans.

Medical literature on the microbiology of kinkajou bites is scarce. We present a patient presenting with hand cellulitis and abscess after a kinkajou bite that grew Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum. We review the literature regarding microbiology and management of kinkajou bites.

IMMUNOHISTOCHEMICAL AND MOLECULAR EVIDENCE OF PUTATIVE NEORICKETTSIA INFECTION IN COATIS ( NASUA NASUA) FROM SOUTHERN BRAZIL.

The pathologic, molecular, and immunohistochemical findings associated with Neorickettsia helminthoeca are described in coatis ( Nasua nasua). Tissue sections (small intestine, lungs, kidney, liver, and spleen) of coatis ( n = 3) that died at the Bela Vista Biological Refuge, Foz do Iguaçu, Paraná, southern Brazil were routinely processed from histopathology. Selected formalin-fixed paraffin-embedded (FFPE) tissue sections of the small intestine, lungs, and spleen were used in an immunohistochemical (IHC) assay designed to identify the antigens of N. helminthoeca. Additionally, FFPE tissue sections of the small intestine were used to demonstrate antigens of canine parvovirus-2 (CPV-2) by IHC. Histopathology revealed chronic enteritis in all coatis. Parasitic enteritis was diagnosed in two coatis; one of these contained examples of a trematode within the lumen of the small intestine and the ovum of a trematode encysted in the intestinal mucosa. Other significant pathologic findings included interstitial pneumonia ( n = 2) and pyogranulomatous splenitis ( n = 1). Positive immunolabeling for N. helminthoeca was identified within macrophages of the small intestine and reticuloendothelial cells within the germinal centers of the spleen of all coatis; the intestinal trematode was N. helminthoeca IHC-positive. All pulmonary sections revealed negative immunolabeling for N. helminthoeca. Furthermore, the antigens of CPV-2 were not identified in the intestine of any coati. These findings indicate that these coatis were infected by N. helminthoeca, but since clinical and gross pathological findings were not recorded, it is uncertain if this pathogen produced clinical disease in this canid host; therefore, coatis may be asymptomatic or dead-end hosts for this organism.

Microscopic and molecular identification of hemotropic mycoplasmas in South American coatis (Nasua nasua).

Hemoplasmas were detected in two apparently healthy captive South American coatis (Nasua nasua) from southern Brazil during an investigation for vector-borne pathogens. Blood was subjected to packed cell volume (PCV) determination, a commercial real-time PCR panel for the detection of Anaplasma spp., Babesia spp., Bartonella spp., Hepatozoon spp., Leishmania spp., Mycoplasma haemofelis, 'Candidatus Mycoplasma turicensis', 'Candidatus Mycoplasma haemominutum', Neorickettsia risticii, Rickettsia rickettsii and Leptospira spp., and a pan-hemoplasma conventional PCR assay. PCV was normal, but both coatis tested positive for hemoplasmas and negative for all the remaining pathogens tested. Using different techniques for microscopy (light, confocal or SEM), structures compatible with hemoplasmas were identified. Sequencing of the 16S rRNA gene identified an organism resembling Mycoplasma haemofelis and another hemotropic Mycoplasma sp., with a sequence identity of 96.8% to a Mycoplasma sp. previously detected in capybaras.

Occurrence and molecular characterization of hemoplasmas in domestic dogs and wild mammals in a Brazilian wetland.

Hemotropic mycoplasmas are known to cause anemia in several mammalian species. The present work aimed to investigate the occurrence of Mycoplasma spp. in wild mammals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil. Between August 2013 and March 2015, 31 Nasua nasua, 78 Cerdocyon thous, seven Leopardus pardalis, 42 dogs, 110 wild rodents, and 30 marsupials were trapped and ectoparasites (ticks and fleas) found parasitizing the animals were collected. Mammals and ectoparasites DNA samples were submitted to conventional PCR assays for Mycoplasma spp. targeting 16S rRNA and RnaseP genes. Twenty-four N. nasua, three C. thous, two domestic dogs, one L. pardalis and one wild rodent were positive for 16S rRNA PCR protocols. Fourteen N. nasua samples were also positive in RnaseP PCR. No marsupial or arthropod showed positivity for Mycoplasma spp. The phylogenetic analyses based on 16S rRNA gene showed that all sequences obtained from dogs, two sequences obtained from C. thous and ten sequences obtained from N. nasua showed to be closely related to Mycoplasma haemocanis/Mycoplasma haemofelis species. Genotypes closely related to 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemomuris were detected in the L. pardalis and in the wild rodent, respectively. Probably a novel Mycoplasma genotype, closely related to a sequence obtained from a Brazilian capybara was detected in 14 N. nasua, based on a concatenated phylogenetic analysis of 16S rRNA and RnaseP genes. The present study revealed that wild animals in southern Pantanal region, Brazil, are exposed to different species of hemoplasmas.

Detection of wild animals as carriers of Leptospira by PCR in the Pantanal biome, Brazil.

Leptospiral infection is widespread in wildlife. In this context, wild ecosystems in tropical countries hold a vast biodiversity, including several species that may act as potential reservoirs of leptospires. The Pantanal biome presents highly favorable environmental conditions for the occurrence of leptospirosis, such as high temperatures, constant flooding, and high biodiversity. The purpose of this study was to detect wild animals as carriers of Leptospira sp. using direct methods (PCR and culture) in the Pantanal biome, Brazil. A total of 35 animals were studied, namely Cerdocyon thous, Nasua nasua, Ozotoceros bezoarticus, and Sus scrofa species. Blood for serology (MAT) and urine for bacteriological culturing and PCR was sampled. The most prevalent serogroups were Javanica and Djasiman. Additionally, 40.6% of these animals presented PCR positive reactions. Seroreactivity associated with the high frequency of leptospiral carriers among the different studied species suggests a high level of exposure of the studied animals to pathogenic Leptospira strains. Our results are still limited and the actual role of the studied animals in the epidemiology of leptospirosis in the Pantanal region remains to be elucidated.

Carriage of Clostridium difficile in free-living South American coati (Nasua nasua) in Brazil.

The objective of this study was to isolate and characterize Clostridium difficile strains in stool samples from a wild urban mammal, a South American coati (Nasua nasua) in Brazil. Forty-six free-living N. nasua were trapped, and stool samples were collected. C. difficile was isolated from three (6.5%) sampled animals, two strains were toxigenic (A+B+CDT-, PCR ribotype 014/020 and 106) and one was non toxigenic (A-B-CDT-, PCR ribotype 053). The present work confirms that ring-tailed coati (N. nasua) could harbor C. difficile strains, including those PCR ribotypes commonly reported in C. difficile infection in humans.

Identification of Encephalitozoon and Enterocytozoon (microsporidia) spores in stool and urine samples obtained from free-living South American Coatis (Nasua nasua).

This study emphasizes the importance of free-living coatis as a potential source of microsporidian infection for humans living in large cities. We found 19 (31.7%) positive results among 60 fecal samples analyzed by PCR-based analysis and the Gram-Chromotrope staining technique (11.7% were positive for Encephalitozoon cuniculi, 6.7% for E. intestinalis, 6.7% for E. hellem, and 6.7% for Enterocytozoon bieneusi). Only 5 (8.4%) urine samples tested positive for E. cuniculi as assessed by the two techniques.

Microbiologia de swabs retais e otológicos em carnívoros silvestres do zoológico do Parque Estadual de Dois Irmãos, Pernambuco / Microbiology of rectal and otologic swabs of wild carnivores from the Zoo of the Parque Estadual de Dois Irmãos, Pernambuco, Brazil

O estudo epidemiológico de patógenos em populações selvagens no ambiente in situ e ex situ é fundamental para implementação de programas de prevenção, controle e monitoramento de enfermidades e para elaboração de políticas públicas de saúde pública e animal. O estudo foi realizado no zoológico do Parque Estadual de Dois Irmãos no período de janeiro a julho de 2011 onde foram coletados swabs anais e otológicos referentes a 29 carnívoros silvestres cativos do zoológico. Dos swabs otológicos analisados, 1/29 (3,4%) foi positivo para Malassezia pachy dermatis no exame direto. No exame microbiológico dos swabs otológicos, observou-se maior freqüência para bactérias do gênero Bacillus 16/29 (55,2%), seguida de Sta phylococcus 15/29 (51,7%), Escherichia coli 7/29 (24,1%), Streptobacillus 1/29 (3,4%), Micrococcus 1/29 (3,4%) e Klebsiella 1/29 (3,4%). Com relação ao exame presuntivo para o gênero Salmonella a partir das amostras de swab retal, observou-se positividade para seis raposas (Cerdocyon thous) e um guaxinim (Procyon cancrivorus). O isolamento de Salmonella spp. em C. thous e em P. cancrivorus indica um risco à saúde pública, principalmente para os profissionais do zoológico que trabalham diretamente com esses animais. Essa pesquisa reforça a importância da criação de estratégias de vigilância epidemiológica voltadas para a prevenção, controle e monitoramento de potenciais reservatórios de agentes etiológicos de doenças infecciosas e parasitárias no ambiente dos zoológicos.

The epidemiological study of pathogens in wild populations in the environment in situ and ex situ is essential for implementation of prevention, control and monitoring of diseases and for development of public policies for public and animal health. The study was conducted at the Zoo of Parque Estadual de Dois Irmãos, Recife/PE, Brazil, from January to July 2011, where we collected anal and otological swabs from 29 Zoo's captive wild carnivores. The analysis of the otological swabs showed that 1/29 (3.4%) were positive for Malassezia pachydermatis on direct examination. In the microbiological examination of otologic swabs we found mainly bacteriae of the genus Bacillus 16/29 (55.2%), followed by Staphylococcus 15/29 (51.7%), Escherichia coli 7/29 (24.1%), Streptobacillus 1/29 (3.4%), Micrococcus 1/29 (3.4%) and Klebsiella 1/29 (3.4%). Regarding the presumptive test for the genus Sal monella from rectal swabs, positive samples were observed in six foxes (Cerdocyon thous) and a raccoon (Procyon cancrivorus). The isolation of Salmonella spp. in C. thous and P. can crivorus indicates risks to public health, especially for zoo professionals who work directly with these animals. This research reinforces the importance of establishing surveillance strategies aimed at prevention, control and monitoring of potential reservoirs of the etiologic agents of infectious and parasitic diseases in the environment of zoos.

Blastomycosis in man after kinkajou bite.

We report transmission of Blastomyces dermatitidis fungal infection from a pet kinkajou to a man. When treating a patient with a recalcitrant infection and a history of an animal bite, early and complete animal necropsy and consideration of nonbacterial etiologies are needed.