Hematological reference values for Alouatta palliata (Primates: Atelidae) and Cebus imitator (Primates: Cebidae) from the Pacific Coast of Costa Rica / Valores de referencia hematológicos para Alouatta palliata (Primates: Atelidae) y Cebus imitator (Primates: Cebidae) de la costa Pacífica de Costa Rica

Abstract Introduction: Wildlife hematological patterns are fundamental for health monitoring, and allows elucidating variations both within and between populations. Among these, hematological parameters are particularly valuable to evaluate the health status of neotropical primate species in the wild. Objective: To define hematological reference values for two species of monkeys in Costa Rica. Methods: During 2014, we collected blood samples from free-ranging mantled howler monkeys, Alouatta palliata (17 females, 18 males) and white-faced capuchin monkeys, Cebus imitator (5 females, 7 males) in seven localities of the Costa Rican Pacific coast. Results: For both species, the hematological values were higher in males, and howler monkey populations differed significantly except for platelets. Conclusions: These hematological values, which differ by sex and locality, will help evaluate the health status of these neotropical primate populations.

Resumen Introducción: Los patrones hematológicos de la vida silvestre son fundamentales para el monitoreo de la salud y permiten dilucidar las variaciones tanto dentro como entre poblaciones. Entre estos, los parámetros hematológicos son particularmente valiosos para evaluar el estado de salud de las especies de primates neotropicales en la naturaleza. Objetivo: Definir valores de referencia hematológicos para dos especies de monos en Costa Rica. Métodos: Durante el 2014 recolectamos muestras de sangre de monos aulladores de manto, Alouatta palliata (17 hembras, 18 machos) y monos capuchinos cariblancos, Cebus imitador (5 hembras, 7 machos) en siete localidades de la costa Pacífica de Costa Rica. Resultados: Para ambas especies, los valores hematológicos fueron mayores en los machos, y las poblaciones de monos aulladores difirieron significativamente con excepción de las plaquetas. Conclusiones: Estos valores hematológicos, que difieren según el sexo y la localidad, ayudarán a evaluar el estado de salud de estas poblaciones de primates neotropicales.

Molecular Epidemiology and Resistance Profile of Campylobacter jejuni and Campylobacter coli Strains Isolated from Different Sources in Brazil.

Aims: The objectives of this study were to genotype a total of 48 Campylobacter jejuni and 39 Campylobacter coli strains isolated in Brazil from 1995 to 2016 by multilocus sequence typing (MLST) and to determine their resistance profile. The presence or points of mutation in the related resistance genes was verified. Results: By MLST, C. jejuni strains were typed into 36 STs and C. coli strains were typed into 27 STs. A total of 70.8% of C. jejuni and 35.9% of C. coli were resistant to at least one antimicrobial tested. The tet(O) gene was detected in 43.7% C. jejuni and in 12.8% C. coli. The ermB gene was not detected and one C. jejuni presented the mutation in the 23S rRNA gene. Besides, 58.3% C. jejuni presented the substitution T86I in the quinolone resistance-determining region of gyrA and 15.4% C. coli presented the substitution T38I. The cmeB gene was detected in 97.9% C. jejuni and in 97.4% C. coli. Conclusion: The presence of C. jejuni and C. coli resistant to some antimicrobial agents of clinical use is of public health concern. The presence of STs shared between Brazilian strains and isolates of different countries is of concern since it might suggest a possible spread of these shared types.

Molecular typing of Campylobacter jejuni strains: comparison among four different techniques.

This study compared the ability of pulsed-field gel electrophoresis (PFGE), flaA small variable region (SVR) sequencing, analysis of the clustered regularly interspaced short palindromic repeats locus by high resolution melting analysis (CRISPR-HRMA), and multilocus sequence typing (MLST) for typing 111 Campylobacter jejuni strains isolated from diverse sources during 20 years in Brazil. For this, we used previous results obtained by PFGE and flaA-SVR sequencing from our research group and performed CRISPR-HRMA and MLST typing for the first time. Furthermore, the discrimination index (DI) of each method was accessed. The DI for PFGE, flaA-SVR sequencing, CRISPR-HRMA, and MLST was 0.980, 0.932, 0.868, and 0.931, respectively. By PFGE and flaA-SVR sequencing, some strains from clinical and non-clinical sources and from humans and animals presented ≥ 80% similarity. Similarly, some strains from different origins presented the same ST and CRISPR-HRMA types. In conclusion, despite the different DI values, all assays provided the same epidemiological information suggesting that a potential transmission may have occurred between C. jejuni from clinical and non-clinical sources and from animals and humans in Brazil. Furthermore it was demonstrated the suitability of PFGE that should be used preferably together with MLST and/or flaA-SVR sequencing for typing C. jejuni strains.

Reversal of Chloroquine Resistance of Plasmodium vivax in Aotus Monkeys.

Chloroquine-resistant (CQR) vivax malaria has emerged as a threat to the malaria elimination agenda. The objective of this study was to assess if a combination of chloroquine (CQ) and prochlorperazine was able to reverse CQ resistance of the Plasmodium vivax AMRU-1 strain from Papua New Guinea in infected Aotus monkeys. For this purpose, in two independent experimental drug efficacy trials, a total of 18 Aotus monkeys infected with blood obtained from donor animals were randomly assigned to treatment and control groups and orally administered CQ at 10 mg/kg or prochlorperazine at 20 mg/kg, alone or in combination, for five consecutive days. Reversal of CQR was achieved in animals that received the drug combination, whereas neither drug alone produced cures. This same drug combination reverses CQR in P. falciparum and could be an alternative for treatment in humans with chloroquine-resistant P. vivax infections.

Molecular detection of fungi of public health importance in wild animals from Southern Brazil.

Some animals have an important relationship with fungal infections, and searching for pathogens in animal samples may be an opportunity for eco-epidemiological research. Since studies involving wildlife are generally restricted, using samples from road kills is an alternative. The aim of this study was to verify whether pathogenic fungi of public health importance occur in wildlife road kills from Santa Catarina State, Brazil. Organ samples (n = 1063) from 297 animals were analysed according to Polymerase Chain Reaction (PCR) using universal primers to detect fungi in general and, subsequently, using primers specific to Paracoccidioides brasiliensis, Histoplasma capsulatum and Cryptococcus spp. There were 102 samples positive for fungal species. Eight samples were positive for P. brasiliensis, three samples were positive for Cryptococcus spp. and one sample had coinfection by these two fungi. No sample was positive for Histoplasma spp. according to the molecular detection. Genetic sequencing allowed the identification of Fungal sp. in 89 samples, Cryptococcus neoformans in two samples and Aspergillus penicillioides in three samples. This study shows the importance of wild animals in the epidemiology of fungal infections and assists in the mapping of pathogen occurrence in a region that was not previously evaluated.

Pathogenic potential and genotypic diversity of Campylobacter jejuni: a neglected food-borne pathogen in Brazil.

Purpose and methodology.Campylobacter jejuni is a major zoonotic pathogen that causes food-borne gastroenteritis worldwide. However, there are only a few studies available that have molecularly characterized C. jejuni strains isolated in Brazil. The aim of this study was to genotype 111 C. jejuni strains isolated from sick humans (43), monkey faeces (19), chicken faeces (14), chicken meat (33) and sewage (2) between 1996 and 2016 in Brazil using flaA-SVR (short variable region) sequencing and PFGE. Furthermore, the presence of 16 virulence genes was analysed by PCR. RESULTS: Using PFGE and flaA-SVR sequencing, the 111 C. jejuni strains studied were grouped into three and two clusters, respectively, and some strains of different origin presented a similarity of ≥80 %. In total, 35 flaA-SVR alleles were detected. Alleles gt45, gt49 and gt57 were the most prevalent, in contrast with those frequently described in the PubMLST database. All 111 C. jejuni strains contained the genes flaA, flhA, cadF, docA, cdtA, cdtB, cdtC, iamA, ciaB, sodB, dnaJ, pldA, racR and csrA. The wlaN gene was detected in 11 strains (9.9 %), and the virB11 in just one strain (0.9 %). CONCLUSIONS: In conclusion, the pathogenic potential of the C. jejuni strains studied was highlighted by the high frequency of the majority of the virulence genes searched. The flaA-SVR sequencing and PFGE results showed that some of the strains studied presented a high genotypic similarity, suggesting potential for transmission between animal sources and humans in this country. Altogether, the results characterize further C. jejuni isolates from Brazil, an important producer and exporter of chicken meat.

Identification of bacterial infection in neotropical primates.

Emerging infectious diseases usually arise from wild animal populations. In the present work, we performed a screening for bacterial infection in natural populations of New World primates. The blood cell bulk DNAs from 181 individuals of four Platyrrhini genera were PCR screened for eubacterial 16S rRNA genes. Bacteria were detected and identified in 13 distinct individuals of Alouatta belzebul, Alouatta caraya, and Cebus apella monkeys from geographically distant regions in the states of Mato Grosso and Pará, Brazil. Sequence analyses showed that these Platyrrhini bacteria are closely related not only to human pathogens Pseudomonas spp. but also to Pseudomonas simiae and sheep-Acari infecting Pseudomonas spp. The identified Pseudomonas possibly represents a group of bacteria circulating in natural monkey populations.

Determinação qualitativa da microbiota bacteriana prepucial e vaginal em macacos-da-noite (Aotus azarai infulatus) criados em cativeiro

O potencial patogênico de diferentes micro-organismos, os relatos de resistência microbiana e a escassez de informações sobre a microbiota do trato genital de primatas não humanos são fatores que justificam a realização deste estudo. O objetivo do estudo foi caracterizar a microbiota bacteriana aeróbica vaginal e prepucial dos macacos-da-noite (Aotus azarai infulatus) criados em cativeiro, avaliando suas frequências entre os sexos e a organização social (acasalados ou não), além de avaliar a sensibilidade antibacteriana. As amostras foram colhidas, de 30 animais adultos e saudáveis, pertencentes ao plantel do Centro Nacional de Primatas, Ananindeua, Pará, através de swabs. Foram semeadas e incubadas em aerobiose em ágar sangue, ágar MacConkey e ágar Chapman, para identificação bacteriana e realização dos testes de sensibilidade através do sistema automatizado VITEK® 2 Compact. Os resultados foram analisados através de estimadores da riqueza e diversidade de espécies, nas comparações foi utilizado o teste qui-quadrado (?2) com (p < 0,05). Entre as cepas gram positivas isoladas a espécie Staphylococcus intermedius foi a de maior frequência entre fêmeas e machos (48,05% e 30,30%, respectivamente). Entre as cepas gram negativas Proteus mirabilis obteve maior frequência em fêmeas (27,10%) e em machos (31,34%). As mesmas espécies também foram as mais isoladas em animais não acasalados (S. intermedius: 41,33% e P. mirabilis: 29,17%) e acasalados (S. intermedius: 38,23% e P. mirabilis: 27,85%). Para as cepas gram positivas o perfil de sensibilidade antimicrobiana in vitro foi o mesmo para ambos os sexos, apresentando 100% de resistência a benzilpenicilina e 100% de sensibilidade a gentamicina, a ciprofloxacina e a norfloxacina. Para as cepas gram negativas, as fêmeas mostraram-se 100% sensíveis somente a levofloxacina e os machos a cinco antibióticos (cefepima, meropenem, amicacina, ciprofloxacina e levofloxacina), ambos os sexos foram menos susceptíveis a nitrofurantoína (fêmeas: 33,70% e machos: 25,40%). A análise dos dados demonstrou que há similaridade entre a composição bacteriana aeróbica da mucosa prepucial e vaginal de A. a. infulatus, que os números de isolamentos e de espécies bacterianas não variam em relação ao sexo e a organização social e que o perfil de sensibilidade antimicrobiana foi semelhante em ambos os sexos para cepas gram positivas e diferente para cepas gram negativas

The pathogenic potential about different micro-organisms, the reports os microbial resistance from the genital treat of not human primates and lack of information are factors that justify the reason for this study. The purpose for this study was to characterize the vaginal and prepuce aerobic microbiota from Nighty Monkeys (Aotus azarai infulatus) raised in captivity, valuating the frequency between sex and social organization (mated or not), besides to evaluate the antibacterial sensibility. The samples were collected from 30 adult and healthy individuals,that are raised on the National Center of Primates, in the municipality of Ananindeua on the State of Pará, Brazil using swabs to do it so. They were sown and incubated in aerobics with blood agar, MacConkey agar and Chapman agar, to bacterial identification and the sensibility tests were done by the automatic system VITEK® 2 Compact. The results were checked using the estimators of richness and diversity of species, on the comparisons were used the qui-squared (Χ2) with (p < 0,05). Between the isolated gram positive strains the Staphylococcus intermedius had the major frequency among females and males (48,05% e 30,30%, respectively). Between the gram negative strains the Proteus mirabilis had major frequency on females (27,10%) and males (31,34%). The same species were the most found in not mated animals (S. intermedius: 41,33% e P. mirabilis: 29,17%) and mated (S. intermedius: 38,23% e P. mirabilis: 27,85%). To the gram positive strains the standard antimicrobial sensibility in vitro were the same for both genders, showing 100% of resistance to benzilpenicilina and 100% de of sensigility against gentamicina, ciprofloxacina and norfloxacina. To the gram negative strains, he females showed 100% of sensibility against levofloxacina and the males against five antibiotics (cefepima, meropenem, amicacina, ciprofloxacina and levofloxacina), both genders were less sensitive to nitrofurantoína (females: 33,70% and males: 25,40%). The data analysis showed that are similiarity between the aerobic bacterial composition of the prepuce and vaginal mucosa from A. a. infulatus, and the numbers of isolations and bacterial species don’t vary related to gender and social organization and the standard antimicrobial sensibility were the same among genders to gram positive and different for gram negative strains.

[Estimation of the survival rate, the relative density and the infection rate of a population of Haemagogus janthinomys Dyar (Diptera, Culicidae) from which strains of yellow fever were isolated in Brazilian Amazon]. / Estimation du taux de survie, de la densité relative et du taux d'infection d'une population d'Haemagogus janthinomys Dyar (Diptera, Culicidae) ayant fourni des souches de fièvre jaune en Amazonie Brésilienne.

The conditions of maintenance of YF virus in brazilian Amazonia are not yet elucidated. Generally, the presence of the virus is attested by human cases of sylvatic origin. During a survey done at the exact place where a man have probably been contaminated, it was possible for the first time in South America, to estimate the mean parity rate of a population of the potential vector Haemagogus janthinomys, from which the YF virus was actually isolated. The survival rate (Ts = 0.96), the biting rate (0.60 mosquitoes/man x hour), and the infection rate (1.71%) were also determinated for the same mosquitoes and have values compatible with the probable conditions of the human contamination. However, more data are needed, in particular in relation with other possible human contaminations and/or circulation of the YF virus in the monkey population (extension and duration of the epizootic episode), in order to know what maintenance cycle is prevalent in this region: a low level transmission, with the mosquito being a "vector-reservoir", or a "constantly moving epizootic wave".

In vitro HIV infection of primate lymphocytes.

Peripheral blood mononuclear cells (PBMC) obtained from four different primate species were tested for their respective ability to support the "in vitro" replication of the human immunodeficiency viruses, HIV-1, and HIV-2. PBMC of Cebus apella, patas (Erythrocebus patas), green (cercopithecus aethiops sabaeus) and rhesus monkeys (Macaca mulatta) were infected "in vitro" with either HIV-1 or HIV-2. Cultures were assayed weekly for particle-associated reverse transcriptase activity. Both viruses were found to be cytolytic for all these monkey's PBMC. Low levels of HIV-1 and HIV-2 infection were observed in Cebus cells. However, productive infection was only detected in HIV-2 infected rhesus PBMC. The capacity of HIV-2 to replicate in rhesus cells may provide a useful model for evaluating antiviral drugs and vaccines.

In vitro HIV infection of primate lymphocytes

El propósito de este estudio fue el de evaluar la capacidad de los virus del SIDA (VIH-1 y VIH-2) para multiplicarse en las células mononuclearres de la sangre periférica (CMSP) de cuatro especies de primates. CMSP de Cebus apella, patas (Erythrocebus patas), monos verdes (cercopithecus aethiops sabeus) y rhesus (Macaca mulatta) fueron infectados "in vitro" con VIH-1 y con VIH-2. La multiplicación de estos virus se determinó midiendo la actividad de la enzima retrotranscriptasa en los cultivos infectados. Ambos virus produjeron efectos citipáticos en dichos cultivos. Se observó un bajo nivel de multiplicación de los virus VIH-1 y VIH-2 en las células provenientes de monos Cebus. Sin embargo, el virus VIH-2 se multiplicó eficientemente en CMSP de monos rhesus. La capacidad que posee el virus de la inmunodeficiencia humana tipo 2, (VIH-2) de multiplicarse en estas células, podría ser utilizada para en la evaluación "in vivo" de productos antivirales y de vacunas

An epidemic of yellow fever in Central Brazil, 1972-1973. II. Ecological studies.

Results of ecological studies undertaken during the 1972-1973 epidemic of yellow fever (YF) in Goiás State, Brazil, suggest that mosquitoes of the genus Haemagogus were the only vectors infected with YF virus. Nine isolations of the agent were made from 1,688 Haemagogus spp. caught within or near forests from 27 January to 3 March 1973. Seven of these isolations cane from mosquitoes collected on the forest floor. No YF virus isolation was made from 791 Haemagogus leucocelaenus or 1,096 Sabethes chloropterus. Haemagogus were caught both in and outside houses located 500 m from the forest, although at a lower rate than in the forest. Haemagogus collection rates in a secondary growth forest were similar to or even higher than those in primary forests. YF virus was recovered from a Cebus monkey and hemagglutination-inhibition and/or neutralizing antibody to YF virus was found in seven (30.4%) of 23 primates examined. There was, however, little evidence of sick or dying monkeys. Among other wild-caught animals only a single marsupial (Caluromys) had antibodies to the virus. These data show that sylvan YF is not confined to the forest, nor always associated with conspicuous epizootics.