Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans / Detecção molecular de protozoários Apicomplexa em tecidos de Alouatta guariba clamitans

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)

O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)

Anatomia do sistema reprodutor feminino de Alouatta belzebul (Linnaeus, 1766) / [Anatomy of the female reproductive system of the Alouatta belzebul (Linnaeus, 1766)]

O conhecimento da anatomia de qualquer animal silvestre é de fundamental importância para sua preservação e proteção. Neste contexto, o presente estudo objetivou descrever a morfologia do sistema reprodutor feminino de Alouatta belzebul. Foram utilizados seis espécimes de A. belzebul, fêmeas, adultas, e livres de lesões. Observou-se macroscopicamente que os ovários têm características morfológicas em formato ovoides, com superfície lisa, e, na análise histológica na região de córtex, evidenciou-se folículos ovarianos em diferentes estágios de desenvolvimento. As tubas uterinas anatomicamente são finas e curvilíneas, apresentando uma camada mucosa, uma muscular e outra serosa. O útero possui formato simples, com fundo globoso, com um miométrio altamente vascularizado, sendo organizado em feixes de fibras musculares lisas. A estrutura anatômica da vagina apresentou-se como um tubo muscular longo de paredes finas, onde, na região vestibular, o óstio externo da uretra é marcado por uma papila uretral bilobada e, na região de vulva, em sua porção caudal, contatou-se um clitóris bem desenvolvido. No que concerne à análise histológica da vagina, verificou-se, em região de mucosa vaginal, um extrato basal composto por epitélio estratificado pavimentoso não queratinizado atrófico. As descrições morfológicas fornecem, de forma inédita, informações importantes relativas à anatomia macroscópica e microscópica do sistema reprodutor feminino dessa espécie.(AU)

Knowledge of the anatomy of any wild animal is of fundamental importance for its preservation and protection. In this context the present study aimed to describe the morphology of the female reproductive system of A. belzebul. We used 6 specimens of A. belzebul, female, adult and free of lesions. It was macroscopically observed that the ovaries are ovoid with smooth surface and the histological analysis in cortical region showed ovarian follicles in different stages of development. The fallopian tubes are anatomically thin and curvilinear, with one mucous layer, one muscular and one serous layer. The uterus was presented in a simple format with a globular fundus, with a highly vascularized myometrium, being organized in bundles of smooth muscle fibers. The anatomical structure of the vagina presented itself as a long thin-walled muscular tube where in the vestibular region the external orifice of the urethra is marked by a bilobed urethral papilla and in the caudal portion in its caudal portion a well-developed clitoris. Regarding the histological analysis of the vagina, a basal extract composed of atrophic non-keratinized stratified squamous epithelium was found in the vaginal mucosa region. The morphological descriptions provide important information regarding the macroscopic and microscopic anatomy of the female reproductive system of this species in an unprecedented way.(AU)

Vascularization of the Alouatta belzebul brain base / Vascularização da base do encéfalo de Alouatta belzebul

We studied the arterial circle in the brain of five specimens of the Alouatta belzebul primate. The material had the arterial system perfused (water at 40°C), injected with stained latex (Neoprene 650), fixed in aqueous formaldehyde solution (10%) and dissected for vessel verification. The arterial circle of this primate is composed of two vascular systems: the vertebra-basilar and the carotid ones, which anastomose to close the arterial circuit. In the caudal portion of the arterial circle, there are the vertebral arteries and their branches: the rostral spinal artery and the caudal inferior cerebellar artery. The anastomosis of the vertebral arteries gives rise to the basilar artery. It presented an anatomical variation at the beginning of its path, forming a double basilar artery, called arterial island. In its course, it emitted branches giving rise to the rostral inferior cerebellar artery, the pontine arteries, the rostral cerebellar arteries, the satellite rostral cerebellar arteries and its terminal branch, the caudal cerebral artery, which presented itself in two segments: the pre-communicating one and post-communicating, joining the internal carotid artery and originating the caudal communicating artery. This group of arteries and anastomoses enclose the caudal portion of the arterial circle. From the right and left internal carotid arteries begins the rostral portion of the arterial circle, which consists of the right and left rostral cerebral arteries and the right and left middle cerebral arteries. The rostral cerebral arteries anastomose into a single trunk, giving rise to the interhemispheric artery, and in A. belzebul and Sapajus libidinosus, the rostral communicating artery is absent. The interhemispheric artery goes to the midbrain region and the corpus callosum knee divides into pericalous artery and callosarginal artery, which will supply the pre and post-central regions of the cerebral hemispheres of this species, as well as other non-human and human primates. It is noted that in the first part of the left rostral cerebral artery, there is a direct inosculation between the recurrent branch of the rostral cerebral artery and left middle cerebral artery to supply the entorhinal region. This fact also occurs in Pongo spp. The middle cerebral artery travels along the lateral sulcus where it emits several superficial branches to irrigate the superior and inferior lateral cortical regions of the frontal, parietal and temporal lobes. It is not part of the arterial circle but is the terminal branch of the internal carotid artery. A. belzebul can be considered to depend on two sources of supply to the brain: the vertebra-basilar and carotid systems, contributing to the intervention of veterinarians during clinical and surgical procedures in other primates, as well as the preservation of wild animals.(AU)

Estudamos o círculo arterial no encéfalo de cinco espécimes do primata Alouatta belzebul. O material teve o sistema arterial perfundido (água a 40°C), injetado com látex corado (Neoprene 650), fixado em solução aquosa de formaldeído (10%) e dissecado para verificação dos vasos. O círculo arterial deste primata é composto por dois sistemas vasculares: vértebro-basilar e o sistema carotídeo, que se anastomosam para fechar o circuito arterial. Na porção caudal do círculo arterial encontra-se as artérias vertebrais e seus ramos: artéria espinal rostral e a cerebelar inferior caudal. A anastomose das artérias vertebrais dá origem a artéria basilar. Esta apresentou uma variação anatômica no início do seu trajeto, formando uma dupla artéria basilar, denominada ilha arterial. Em seu trajeto emitiu ramos dando origem a artéria cerebelar inferior rostral, as artérias pontinas, as artérias cerebelares rostrais, as artérias cerebelares rostrais satélites e o seu ramo terminal, a artéria cerebral caudal, que apresentou-se em dois segmentos: o pré-comunicante e pós-comunicante, unindo-se a artéria carótida interna e originando a artéria comunicante caudal. Este grupo de artérias e anastomoses encerram a porção caudal do círculo arterial. Das artérias carótidas internas direita e esquerda, inicia-se a porção rostral do círculo arterial, ao qual é constituído pelas artérias cerebrais rostrais direita e esquerda e as artérias cerebrais médias direita e esquerda. As artérias cerebrais rostrais se anastomosam em um tronco único dando origem a artéria inter-hemisférica e em A. belzebul e Sapajus libidinosus, a artéria comunicante rostral se encontra ausente. A artéria inter-hemisférica segue para região média do encéfalo e no joelho do corpo caloso se divide em artéria pericalosa e artéria calosomarginal, que vão suprir as regiões pré e pós-central dos hemisférios cerebrais desta espécie, assim como outros primatas não humanos e humano. Nota-se que na primeira parte da artéria cerebral rostral esquerda, ocorre uma inosculação direta entre o ramo recorrente da artéria cerebral rostral e artéria cerebral média esquerda para suprir a região entorrinal, esse fato também ocorre em Pongo spp. A artéria cerebral média segue seu trajeto pelo sulco lateral onde emite vários ramos superficiais para irrigar as regiões corticais supero e ínfero lateral do lobo frontal, parietal e temporal, esta não faz parte do círculo arterial mas é o ramo terminal da artéria carótida interna. Pode-se considerar que A. belzebul depende de duas fontes de suprimento para o encéfalo: os sistemas vértebro-basilar e carotídeo, contribuindo na intervenção de médicos veterinários durante os procedimentos clínicos e cirúrgicos em outros primatas, assim como na preservação de animais silvestres.(AU)

Combination of surveillance tools reveals that Yellow Fever virus can remain in the same Atlantic Forest area at least for three transmission seasons

BACKGROUND In Brazil, the Yellow Fever virus (YFV) is endemic in the Amazon, from where it eventually expands into epidemic waves. Coastal south-eastern (SE) Brazil, which has been a YFV-free region for eight decades, has reported a severe sylvatic outbreak since 2016. The virus spread from the north toward the south of the Rio de Janeiro (RJ) state, causing 307 human cases with 105 deaths during the 2016-2017 and 2017-2018 transmission seasons. It is unclear, however, whether the YFV would persist in the coastal Atlantic Forest of RJ during subsequent transmission seasons. OBJECTIVES To conduct a real-time surveillance and assess the potential persistence of YFV in the coastal Atlantic Forest of RJ during the 2018-2019 transmission season. METHODS We combined epizootic surveillance with fast diagnostic and molecular, phylogenetic, and evolutionary analyses. FINDINGS Using this integrative strategy, we detected the first evidence of YFV re-emergence in the third transmission season (2018-2019) in a dying howler monkey from the central region of the RJ state. The YFV detected in 2019 has the molecular signature associated with the current SE YFV outbreak and exhibited a close phylogenetic relationship with the YFV lineage that circulated in the same Atlantic Forest fragment during the past seasons. This lineage circulated along the coastal side of the Serra do Mar mountain chain, and its evolution seems to be mainly driven by genetic drift. The potential bridge vector Aedes albopictus was found probing on the recently dead howler monkey in the forest edge, very close to urban areas. MAIN CONCLUSIONS Collectively, our data revealed that YFV transmission persisted at the same Atlantic Forest area for at least three consecutive transmission seasons without the need of new introductions. Our real-time surveillance strategy permitted health authorities to take preventive actions within 48 h after the detection of the sick non-human primate. The local virus persistence and the proximity of the epizootic forest to urban areas reinforces the concern with regards to the risk of re-urbanisation and seasonal re-emergence of YFV, stressing the need for continuous effective surveillance and high vaccination coverage in the SE region, particularly in RJ, an important tourist location.

Plasmodium infection and its association with biochemical and haematological parameters in free-living Alouatta guariba clamitans (Cabrera, 1940) (Primates: Atelidae) in Southern Brazil

BACKGROUND The influence of Plasmodium spp. infection in the health of Southern brown howler monkey, Alouatta guariba clamitans, the main reservoir of malaria in the Atlantic Forest, is still unknown. OBJECTIVES The aim of this study was to investigate the positivity rate of Plasmodium infection in free-living howler monkeys in an Atlantic Forest fragment in Joinville/SC and to associate the infection with clinical, morphometrical, haematological and biochemical alterations. METHODS Molecular diagnosis of Plasmodium infection in the captured monkeys was performed by Nested-polymerase chain reaction (PCR) (18S rRNA and coxI). Haematological and biochemical parameters were compared among infected and uninfected monkeys; clinical and morphometrical parameters were also compared. FINDINGS The positivity rate of Plasmodium infection was 70% among forty captured animals, the highest reported for neotropical primates. None statistical differences were detected in the clinical parameters, and morphometric measures comparing infected and uninfected groups. The main significant alteration was the higher alanine aminotransferase (ALT) levels in infected compared to uninfected monkeys. MAIN CONCLUSIONS Therefore, Plasmodium infection in howler monkeys may causes haematological/biochemical alterations which might suggest hepatic impairment. Moreover, infection must be monitored for the eco-epidemiological surveillance of malaria in the Atlantic Forest and during primate conservation program that involves the animal movement, such as translocations.

Aspectos ultrassonográficos, biometria e dopplerfluxometria ocular do bugio ruivo (Alouatta fusca) / Ultrasonography aspects, biometry and Doppler of howler monkey (Alouatta fusca) ocular bulb

Este estudo teve como objetivo geral descrever os aspectos ultrassonográficos normais das estruturas do bulbo ocular de primatas da espécie Alouatta fusca, determinando ainda os valores da biometria ocular, e os parâmetros da dopplerfluxometria ocular das artérias oftálmica interna e central da retina. Vinte exames ultrassonográficos oculares foram realizados em dez primatas da espécie Alouatta fusca clinicamente saudáveis, sem sinais de doença ocular ao exame oftalmológico. Procedeu-se a descrição ultrassonográfica das estruturas oculares e posteriormente a biometria foi obtida em quatro distâncias distintas: (D1) correspondentes à câmara anterior, (D2) espessura da lente, (D3) profundidade da câmara vítrea e (D4) comprimento axial do bulbo ocular. Na dopplerfluxometria foram avaliadas as artérias oftálmica interna e central da retina quanto ao índice de resistividade (IR), de pulsatilidade (IP), e as velocidades do pico sistólico (VPS) ediastólica final (VDF). Os valores de biometria foram submetidos a teste de comparação quanto ao gênero dos animais e cortes ultrassonográficos, utilizando o teste t de Student. O mesmo teste foi realizado para comparação dos resultados de dopplerfluxometria entre fêmeas e machos. A descrição ultrassonográfica das estruturas oculares e vascularização do bugio ruivo mostraram-se semelhantes a espécies como o cão, o gato e o homem. Os valores biométricos médios encontrados foram de 2,1±0,38 mm para D1, 3,7±0,30mm para D2, 10,4±0,78mm para D3 e 19,3±1,64mm para D4. Os valores de dopplerfluxometria da artéria oftálmica e da artéria central da retina foram respectivamente: VPS de 25,6cm/s e 14,6cm/s; VDF de 15,8cm/s e 10,7cm/s; IR de 0,7 e 0,5; IP de 1,4 e 0,8.(AU)

This study aimed to describe the sonographic features of normal ocular structures, the ocular biometry and Doppler parameters of the internal ophthalmic artery and central retinal artery of the Alouatta fusca. Twenty ocular ultrasonographic examinations were perform in ten primate species of the Alouatta fusca. Proceeded to the sonographic description of the ocular structures and later biometrics was obtained in four distances: (D1) corresponding to the anterior chamber (D2) lens thickness (D3) vitreous chamber and (D4) axial length of the eyeball. Doppler ultrasound evaluated internal ophthalmic artery and central retinal artery as for the resistivity Index (RI), pulsatility index (PI), peak systolic velocity (PSV) and end diastolic velocity (EDV). The values of biometrics and flowmetry underwent comparison test between genders and sonographic views, by Student t test. The anatomical sonographic description of the ocular structures and vasculature of the red howler were similar to species such as dog, cat and man. The biometric average values found were 2.1±0.38mm for D1, 3.7±0.30mm for D2, 10.4±0.78mm for D3 and 19.3±1.64mm for D4. The Doppler values of the ophthalmic artery and central retinal artery found were: PSV of 25.6cm/s and 14.6cm/s; VDF 15.8cm/s and 10.7cm/s; IR 0.7 and 0.5; IP 1.4 and 0.8.(AU)

Bloqueio de plexo braquial em um bugio-ruivo (Alouatta guariba): relato de caso / Brachial plexus block in a red howler monkey (Alouatta guariba): case report

Os bloqueios regionais vêm sendo explorados e difundidos no dia a dia da medicina veterinária. O presente trabalho relata a execução do bloqueio de plexo braquial em um bugio-ruivo (Alouatta guariba) submetido à amputação de três dígitos. Foi utilizada ropivacaína para o bloqueio, com o paciente sob anestesia geral, com auxílio de neuroestimulador periférico. Quatro horas após a cirurgia e cinco horas após o bloqueio, o paciente não apresentava desconforto ou evitava a manipulação do membro operado e não foram observadas complicações. O bloqueio do plexo braquial é mais comumente utilizado quando se pretende promover analgesia e relaxamento muscular do membro torácico em procedimentos cirúrgicos distais à articulação escápulo-umeral. Há carência em estudos anatômicos sobre diversos primatas, incluindo o bugio-ruivo (Alouatta guariba). Apesar da crescente expansão da clínica de animais silvestres, ainda há lacunas no que diz respeito à execução de técnicas anestésicas em diversas espécies.(AU)

The regional blocks are being exploited and disseminated on a daily basis of veterinary medicine. This study describes the implementation of the brachial plexus block in a red howler-monkey (Alouatta guariba) that underwent amputation of three digits. Ropivacaine was employed for blocking, with the patient under general anesthesia, with the aid of a peripheral nerve stimulator. Four hours after surgery, and five hours after the blockade, the patient had no discomfort nor avoided manipulation of the operated limb and no complications were observed. The brachial plexus block is most commonly used when you want to promote analgesia and muscle relaxation of the forelimb in distal surgical procedures to the scapular-humeral joint. There is a lack of anatomical studies on various primates, including the red howler-monkey (Alouatta guariba). Despite the growing expansion of clinic of wild animals, there are still shortcomings with regards to the implementation of anesthetic techniques in many species.(AU)

Genome analysis of yellow fever virus of the ongoing outbreak in Brazil reveals polymorphisms

The current yellow fever outbreak in Brazil is the most severe one in the country in recent times. It has rapidly spread to areas where YF virus (YFV) activity has not been observed for more than 70 years and vaccine coverage is almost null. Here, we sequenced the whole YFV genome of two naturally infected howler-monkeys (Alouatta clamitans) obtained from the Municipality of Domingos Martins, state of Espírito Santo, Brazil. These two ongoing-outbreak genome sequences are identical. They clustered in the 1E sub-clade (South America genotype I) along with the Brazilian and Venezuelan strains recently characterised from infections in humans and non-human primates that have been described in the last 20 years. However, we detected eight unique amino acid changes in the viral proteins, including the structural capsid protein (one change), and the components of the viral replicase complex, the NS3 (two changes) and NS5 (five changes) proteins, that could impact the capacity of viral infection in vertebrate and/or invertebrate hosts and spreading of the ongoing outbreak.

Detection of Plasmodium in faeces of the New World primate Alouatta clamitans

Abstract Plasmodium falciparum and Plasmodium vivax have evolved with host switches between non-human primates (NHPs) and humans. Studies on the infection dynamics of Plasmodium species in NHPs will improve our understanding of the evolution of these parasites; however, such studies are hampered by the difficulty of handling animals in the field. The aim of this study was to detect genomic DNA of Plasmodium species from the faeces of New World monkeys. Faecal samples from 23 Alouatta clamitans from the Centre for Biological Research of Indaial (Santa Catarina, Brazil) were collected. Extracted DNA from faecal samples was used for molecular diagnosis of malaria by nested polymerase chain reaction. One natural infection with Plasmodium simium was identified by amplification of DNA extracted from the faeces of A. clamitans. Extracted DNA from a captive NHP was also used for parasite genotyping. The detection limit of the technique was evaluated in vitro using an artificial mixture of cultured P. falciparum in NHP faeces and determined to be 6.5 parasites/µL. Faecal samples of New World primates can be used to detect malaria infections in field surveys and also to monitor the genetic variability of parasites and dynamics of infection.

Distribución geográfica del mono aullador rojo ( Alouatta seniculus) y la fiebre amarilla en Colombia / Geographical distribution of the red howler monkey ( Alouatta seniculus ) and yellow fever in Colombia

Introducción. Colombia es un país con gran diversidad de primates no humanos, entre los cuales se destaca el mono aullador rojo ( Alouatta seniculus ) por su distribución y el papel que desempeña en la presentación de la fiebre amarilla. Objetivo. Describir la coincidencia geográfica del hábitat del mono aullador rojo y la presencia de fiebre amarilla. Materiales y métodos. Se hizo un estudio de carácter descriptivo. Los antecedentes de la fiebre amarilla en Colombia se obtuvieron de los informes y boletines del Instituto Nacional de Salud y del estudio de 2013 de Segura, et al. La presencia de A. seniculus se determinó con base en la plataforma Global Biodiversity Information Facility y el Sistema de Información sobre Biodiversidad de Colombia; los mapas se elaboraron con el programa Diva-Gis, y el modelo de nicho ecológico bajo las condiciones actuales, con el programa Maxent. Resultados. Los departamentos con mayor presencia de A. seniculus fueron Antioquia, Meta y Casanare; en 69,5 % de los departamentos con antecedentes de notificación de fiebre amarilla también había A. seniculus. El modelo de nicho ecológico evidenció que Antioquia, Bolívar, La Guajira, Magdalena, Meta, Santander, Norte de Santander y Vichada tenían porciones de territorio con un índice de probabilidad cercano a 0,9 (90 %). Conclusiones. En 69,5 % de los departamentos con antecedentes de fiebre amarilla se registró la presencia de A. seniculus , lo cual resulta relevante por el papel que los primates no humanos desempeñan como reservorio natural del virus y por su contribución en la presentación de la fiebre amarilla, lo cual les confiere gran utilidad como centinelas.

Introduction: Colombia is a country with an important diversity of non-human primates, of which the red howler monkey ( Alouatta seniculus ) stands out because of its distribution and the role it plays in the occurrence of yellow fever. Objective: To describe the geographic co-occurrence of Alouatta seniculus and the reported presence of yellow fever. Materials and methods: We conducted a descriptive study. The reported presence of yellow fever in Colombia was obtained from the reports and bulletins issued by the Instituto Nacional de Salud , and the study by Segura, et al . (2013). The occurrence of A. seniculus was determined based on the data from the Global Biodiversity Information Facility and the Colombian Biodiversity Information System. A map of the occurrence was developed using the DIVA-GIS program, and the ecological niche model under current conditions was created with the Maxent program. Results: The departments with the highest occurrence of A. seniculus were Antioquia, Meta and Casanare; 69.5% of the departments with reported history of yellow fever had co-occurrence with A. seniculus . The ecological niche model showed that Antioquia, Bolívar, La Guajira, Magdalena, Meta, Santander, Norte de Santander and Vichada had geographical portions with a probability rate nearing to 0.9 (90%). Conclusions: In 69.5% of the departments with a history of yellow fever there was co-occurrence with A. seniculus , which is relevant because non-human primates play a well-known role as natural reservoirs of the virus, and they might contribute to the occurrence of the yellow fever, which makes them very useful as sentinels.

Frequency of histopathological changes in Howler monkeys ( Alouatta sp. ) naturally infected with yellow fever virus in Brazil

Abstract: INTRODUCTION: Due to the importance that Howler monkeys have on the yellow fever (YF) epidemiological sylvatic cycle in Brazil, more accurate morphological diagnostic criteria needs to be established, especially considering the differences that may exist between the genera of Brazilian non-human primates (NHPs) involved in yellow fever virus (YFV) epizootics. METHODS: Records of YF epizootics in NHPs in Brazil between 2007 and 2009 were obtained from the Brazilian Ministry of Health database to select YF positive (n=98) Howler monkeys (Alouatta sp.) for this study. The changes described in the histopathological reports were categorized by organ and their frequencies calculated. RESULTS: The most frequent lesions observed in the animals with YF were hepatocyte apoptosis (Councilman body formation), midzonal hepatocyte necrosis, steatosis, liver hemorrhage, inflammatory mononuclear cell infiltration of the liver, renal acute tubular necrosis and interstitial nephritis. Midzonal hepatocyte necrosis, steatosis and hemorrhage presented positive correlations with apoptosis of hepatocytes, suggesting strong YFV pathogenic effect association; they were also the main histopathological changes in the Alouatta sp. A pronounced negative correlation between apoptosis of hepatocytes and hepatic mononuclear cell infiltration pointed to significant histopathological differences between YFV infection in Howler monkeys and humans. CONCLUSIONS: The results warn that NHPs may exhibit different response patterns following YFV infection and require a more careful diagnosis. Presumptive diagnosis based on primate histopathological lesions may contribute to public health service control.

Epizootic amebiasis outbreak in wild black howler monkeys (Alouatta caraya) in a wildlife facility during captivity prior to translocation - Recommendations to wildlife management programs / Surto epizoótico de amebíase em Bugio preto (Alouatta caraya) provenientes de vida livre durante o cativeiro prévio a translocação - Recomendações a programas de manejo de fauna

The capture and quarantine of non-human primates could be necessary in some circumstances including those required under environmental permit. Mortality is undesirable for wildlife management programs and could be related to opportunistic pathogens, for example, deaths due to intestinal protozoa infection outbreaks as described here. Parasitological, necroscopic, microbiological, and molecular tests were used in the diagnosis of severe necrotic enteritis leading to death of three female and two male black howler monkeys (Alouatta caraya) captured and held in quarantine prior to translocation. Parasitological tests showed the presence of cysts of Entamoeba histolytica/dispar (5/5), Entamoeba coli (5/5) and Giardia duodenalis (1/5). Necroscopic assessment revealed areas of severe multifocal necrosis in the intestinal mucosa and submucosa. Histopathological examination revealed the presence of structures morphologically compatible to Entamoeba spp. in all individuals examined. Furthermore, G. duodenalis (1/5) was demonstrated by the nested PCR technique. During temporary captivity of non-human primates in management programs, proper handling protocols, including fast or immediate destination, are suggested in order to mitigate the negative effects of stress and decrease the risk of infections.(AU)

Óbitos por infecções causadas por protozoários intestinais são indesejáveis para programas de manejo de fauna, exigidos no âmbito do licenciamento ambiental e que demandam a captura e quarentena de primatas não humanos. Exames coproparasitológicos, necroscópicos, microbiológicos e moleculares foram utilizados para a confirmação do diagnóstico de uma severa enterite necrótica que levou a óbito três fêmeas e dois machos de Alouatta (A.) caraya capturados e mantidos em quarentena prévia à translocação. Exames coproparasitológicos revelaram a presença de cistos de Entamoeba (E.) histolytica/dispar (5/5), Entamoeba (E.) coli (5/5) e Giardia (G.) duodenalis (1/5). A avaliação necroscópica revelou áreas de necrose multifocal severa na mucosa e submucosa intestinal. A análise microscópica revelou a presença de estruturas morfologicamente compatíveis com trofozoítos do gênero Entamoeba spp. em todos os indivíduos examinados. Além disso, G. duodenalis (1/5) foi demonstrada pela técnica de Nested PCR. Sugere-se que quando for necessário o cativeiro temporário de primatas, deverão ser adotados protocolos de manejo adequados buscando a destinação imediata dos animais, de moto a mitigar os efeitos negativos do estresse e reduzir o risco da ocorrência de infecções.(AU)

Yellow fever impact on brown howler monkeys (Alouatta guariba clamitans) in Argentina: a metamodelling approach based on population viability analysis and epidemiological dynamics

In South America, yellow fever (YF) is an established infectious disease that has been identified outside of its traditional endemic areas, affecting human and nonhuman primate (NHP) populations. In the epidemics that occurred in Argentina between 2007-2009, several outbreaks affecting humans and howler monkeys (Alouatta spp) were reported, highlighting the importance of this disease in the context of conservation medicine and public health policies. Considering the lack of information about YF dynamics in New World NHP, our main goal was to apply modelling tools to better understand YF transmission dynamics among endangered brown howler monkey (Alouatta guariba clamitans) populations in northeastern Argentina. Two complementary modelling tools were used to evaluate brown howler population dynamics in the presence of the disease: Vortex, a stochastic demographic simulation model, and Outbreak, a stochastic disease epidemiology simulation. The baseline model of YF disease epidemiology predicted a very high probability of population decline over the next 100 years. We believe the modelling approach discussed here is a reasonable description of the disease and its effects on the howler monkey population and can be useful to support evidence-based decision-making to guide actions at a regional level.

Evaluation of arboviruses of public health interest in free-living non-human primates (Alouatta spp., Callithrix spp., Sapajus spp.) in Brazil

INTRODUCTION: The aim of the present study was to evaluate the presence of arboviruses from the Flavivirus genus in asymptomatic free-living non-human primates (NHPs) living in close contact with humans and vectors in the States of Paraná and Mato Grosso do Sul, Brazil. METHODS: NHP sera samples (total n = 80, Alouatta spp. n = 07, Callithrix spp. n = 29 and Sapajus spp. n = 44) were screened for the presence of viral genomes using reverse transcription polymerase chain reaction and 10% polyacrylamide gel electrophoresis techniques. RESULTS: All of the samples were negative for the Flavivirus genome following the 10% polyacrylamide gel electrophoresis analysis. CONCLUSIONS: These negative results indicate that the analyzed animals were not infected with arboviruses from the Flavivirus genus and did not represent a risk for viral transmission through vectors during the period in which the samples were collected. .

Avaliação ultrassonográfica e mensurações das glândulas adrenais em primatas não humanos neotropicais: mico-de-cheiro (Saimiri sciureus), macaco-da-noite (Aotus azarae infulatus) e bugio-ruivo (Alouatta guariba clamitans) / Adrenal glands measurements and ultrasound evaluation in nonhuman primates: squirrel monkey (Saimiri sciureus), owl monkey (Aotus azarae infulatus) and howler monkey (Alouatta guariba clamitans)

As glândulas adrenais possuem funções endócrinas relacionadas a múltiplas funções vitais, estando intimamente relacionadas à capacidade do animal em se adaptar ao estresse. O exame ultrassonográfico é o método diagnóstico de escolha para avaliação das glândulas em diferentes espécies. Considerando a escassa literatura, questiona-se se as doenças adrenais em primatas não humanos são incomuns ou subdiagnosticadas, havendo a hipótese desse fato ser determinado pela falta de parâmetros. Objetivou-se descrever as características ultrassonográficas das glândulas adrenais para três espécies de primatas não humanos mantidas em cativeiro: Saimiri sciureus (mico-de-cheiro), Aotus azarae infulatus (macaco-da-noite) e Alouatta guariba clamitans (bugio-ruivo). Conclui-se que é possível a identificação das glândulas adrenais por meio de exame ultrassonográfico, sendo que os padrões de referência foram estabelecidos com sucesso para as espécies em questão. Ressalta-se que a adequação de animais em ambientes estressantes é frequentemente acompanhada por uma hipertrofia das glândulas adrenais, portanto deve-se levar em consideração que as mensurações realizadas nesse estudo foram estabelecidas em animais de cativeiro...

The adrenal glands have endocrine functions related to multiple vital functions and are closely related to the animal's ability to adapt to stress. The ultrasound is the diagnostic method of choice for evaluation of glands in different species. Considering the scarce literature, one may question whether the adrenal disorders in nonhuman primates are uncommon or underdiagnosed, and a hypothesis exists that this fact is determined by the lack of parameters. The goal is to describe the sonographic features of the adrenal glands for three species of nonhuman primates kept in captivity: squirrel monkey (Saimiri sciureus), owl monkey (Aotus azarae infulatus) and howler monkey (Alouatta guariba clamitans). It is concluded that it is possible to identify the adrenal glands by ultrasound, and the reference standards have been established successfully for the species in question. It is noteworthy that the adaptation of animals in many stressful environments is often accompanied by a hypertrophy of the adrenal glands, so one should take into account that the measurements performed in this study were established in captive animals...

Plasmodium simium/Plasmodium vivax infections in southern brown howler monkeys from the Atlantic Forest

Blood infection by the simian parasite, Plasmodium simium, was identified in captive (n = 45, 4.4%) and in wild Alouatta clamitans monkeys (n = 20, 35%) from the Atlantic Forest of southern Brazil. A single malaria infection was symptomatic and the monkey presented clinical and haematological alterations. A high frequency of Plasmodium vivax-specific antibodies was detected among these monkeys, with 87% of the monkeys testing positive against P. vivax antigens. These findings highlight the possibility of malaria as a zoonosis in the remaining Atlantic Forest and its impact on the epidemiology of the disease.

Ultraestrutura comparativa da língua do sagui-de-tufo-preto (Callithrix penicillata) e do bugio-preto (Alouatta caraya) em diferentes faixas etárias / Comparative ultrastructure of the tongue of black-tufted-ear-marmoset (Callithrix penicillata) and black-howler-monkey (Alouatta caraya) in different age groups

Para a análise e descrição comparativa da morfologia da língua e de suas papilas, bem como a distribuição destas, foram utilizados três animais de faixas etárias distintas (filhote, jovem, adulto), de duas espécies de primatas, Callithrix penicillata (sagui-de-tufo-preto) e Alouatta caraya (bugio-preto), ambas sendo observadas através da microscopia eletrônica de varredura. Os animais da espécie Callithrix penicillata eram procedentes de um criadouro comercial de animais selvagens em Atibaia/SP e vieram a óbito por causas naturais, e os da espécie Alouatta caraya eram provenientes do acervo do Laboratório de Anatomia Macroscópica da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP). Com o material analisado e a técnica empregada pôde-se constatar que as línguas dos saguis-de-tufo-preto e dos bugios-pretos analisadas apresentavam papilas linguais similares às descritas para outras espécies de primatas. As línguas possuíam diferenças morfológicas, principalmente para alguns tipos de papilas linguais, em detrimento do avançar da idade dos animais. De forma geral, foram observados quatro tipos de papilas linguais, sendo estas: filiforme, fungiforme, valada e folhada. As línguas dos bugio-pretos apresentavam características de animais com dietas herbívoras. Possuindo desta forma papilas filiformes com formatos variados (coroa, lança, multifilamentar), de acordo com as regiões da língua, e também robustas papilas, principalmente na proeminência lingual. Já as línguas dos saguis-de-tufo-preto possuíam características de animais onívoros. Com papilas filiformes em formato de coroa, grandes quantidades de papilas fungiformes e desenvolvidas papilas folhadas. Observou-se que as modificações das papilas linguais que ocorrem nos animais após o nascimento apresentaram correlação com a mudança nos tipos de alimentos consumidos. Sendo presumível então concluir que as dietas possuem relação direta para com as alterações morfológicas e estruturais das papilas linguais nos animais ora aqui analisados.

For the comparative analysis and description of the morphology of the tongue and taste buds as well as the distribution of these we have used three animals of different age groups (puppy, young adult), of two species of primate, Callithrix penicillata (black-tufted-ear-marmoset) and Alouatta caraya (black-howler-monkey), both being observed by scanning electron microscopy. The animals of the species Callithrix penicillata were from a commercial breeding of wild animals in Atibaia-SP and they had died of natural causes, and the specie Alouatta caraya were from the collection of Macroscopic Anatomy Laboratory of Faculty of Veterinary Medicine and Animal Science University of São Paulo (FMVZ-USP). With the material analyzed and the technique employed could be seen that the tongue of black-tufted-ear-marmosets and black-howler-monkeys had examined lingual papillae similar to those described for other primate species. The tongue has morphological differences, especially for some types of lingual papillae, at the expense of advancing age of the animals. Overall, we found four types of lingual papillae, which are: filiform, fungiform, vallate and litter. The tongues of the black-howler-monkey had characteristics of animals with herbivorous diets. Possessing this way with the filiform papillae varied formats (crown, spear, multifilament), according to the regions of the tongue, and also robust taste, especially in the lingual prominence. Since the tongues of the black-tufted-ear-marmosets owned features of omnivorous animals. With filiform-shaped crown, large amounts of fungiform papillae and foliate papillae developed. It was observed that the changes of tongue papillae that occur in animals after birth correlated with the change in the types of foods consumed. Being suspected then conclude that diets have direct relation to the morphological and structural of the lingual papillae now here in the animals analyzed.

Detection of arboviruses of public health interest in free-living New World primates (Sapajus spp.; Alouatta caraya) captured in Mato Grosso do Sul, Brazil

Introduction A sero-epidemiological survey was undertaken to detect the circulation of arboviruses in free-living non-human primates. Methods Blood samples were obtained from 16 non-human primates (13 Sapajus spp. and three Alouatta caraya) that were captured using terrestrial traps and anesthetic darts in woodland regions in the municipalities of Campo Grande, Aquidauana, Jardim, Miranda and Corumbá in the State of Mato Grosso do Sul, Brazil. The samples were sent to the Instituto Evandro Chagas (IEC) in Ananindeua, Pará, Brazil, to detect antibodies against 19 species of arboviruses using a hemagglutination inhibition test (HI). Results Of the 16 primates investigated in the present study, five (31.2%) were serologically positive for an arbovirus. Of these five, two (12.5%) exhibited antibodies to the Flavivirus genus, one (6.2%) exhibited a monotypic reaction to Cacipacoré virus, one (6.2%) was associated with Mayaro virus, and one (6.2%) was positive for Oropouche virus. Conclusions Based on the positive serology observed in the present study, it was possible to conclude that arboviruses circulate among free-living primates. The viruses in the areas studied might have been introduced by infected humans or by primates from endemic or enzootic areas. Studies of this nature, as well as efficient and continuous surveillance programs, are needed to monitor viral activities in endemic and enzootic regions. .

First report of yellow fever virus in non-human primates in the State of Parana, Brazil

Sylvatic yellow fever is a zoonosis associated mainly with wild animals, especially those in the genus Alouatta, that act as the source of infection. Once infected, these animals pass the disease on to humans by way of an infected mosquito belonging to the genera Aedes, Haemagogus, or Sabethes. The present study is the first report of a case of yellow fever in non-human primates (NHP) in the State of Paraná, Brazil. After the case was diagnosed, several prophylactic measures were adopted to prevent outbreaks of the disease in humans.