Hematological and serum biochemistry evaluation in howler monkeys (Alouatta caraya) and capuchin monkeys (Sapajus apella): A comparative study.

BACKGROUND: Evaluation of blood parameters in captive non-human primates (NHPs) is crucial for monitoring their health and ensuring that their environment meets their physiological requirements. METHODS: We performed hemogram, serum biochemistry, and parasitological exams in 20 howler monkeys and 21 capuchin monkeys. RESULTS: In both species, over 50% of the individuals presented at least one parasite. There was a negative effect of age on red blood cell (RBC), white blood cell, platelets, total protein, globulin, and alkaline phosphatase, and a positive effect on the A:G ratio, gamma-glutamyl transferase, and mean platelet volume (MPV). Capuchin monkeys presented the highest platelets and alanine aminotransferase (ALT) values and howler monkeys presented the highest MPV, aspartate aminotransferase, ALT, amylase, glucose, bilirubin, and triglycerides values. We observed an interaction between species and sex on RBC, Htc, mean corpuscular hemoglobin concentration, and cholesterol. CONCLUSIONS: Species differences found in blood parameters may reflect differences in physiological adaptations associated with ecological and morphological traits and are clinically relevant for evaluating animal health and the suitability of breeding programs.

Zika Virus Infection Damages the Testes in Pubertal Common Squirrel Monkeys (Saimiri collinsi).

During the Zika virus (ZIKV) outbreak and after evidence of its sexual transmission was obtained, concerns arose about the impact of the adverse effects of ZIKV infection on human fertility. In this study, we evaluated the clinical-laboratory aspects and testicular histopathological patterns of pubertal squirrel monkeys (Saimiri collinsi) infected with ZIKV, analyzing the effects at different stages of infection. The susceptibility of S. collinsi to ZIKV infection was confirmed by laboratory tests, which detected viremia (mean 1.63 × 106 RNA copies/µL) and IgM antibody induction. Reduced fecal testosterone levels, severe testicular atrophy and prolonged orchitis were observed throughout the experiment by ultrasound. At 21 dpi, testicular damage associated with ZIKV was confirmed by histopathological and immunohistochemical (IHC) analyses. Tubular retraction, the degeneration and necrosis of somatic and germ cells in the seminiferous tubules, the proliferation of interstitial cells and an inflammatory infiltrate were observed. ZIKV antigen was identified in the same cells where tissue injuries were observed. In conclusion, squirrel monkeys were found to be susceptible to the Asian variant of ZIKV, and this model enabled the identification of multifocal lesions in the seminiferous tubules of the infected group evaluated. These findings may suggest an impact of ZIKV infection on male fertility.

Effect of age and sex in renal function by ultrasound and serum chemistry in two primate species (Alouatta caraya and Sapajus apella).

BACKGROUND: Comparative studies of kidney morphophysiology in nonhuman primates can help us investigate interspecies differences in growth and aging patterns. METHODS: We tested the effect of age and sex in renal morphophysiology in 21 Alouatta caraya and 21 Sapajus apella (age range = 0.5-26 years) by ultrasound, red blood cell (RBC) count, and kidney function tests. RESULTS: A. caraya had greater growth rate and absolute renal volume than S. apella, but the latter showed a greater relative renal volume and RBC count. There was a negative relationship between RBC and age, a positive relationship between creatinine and body mass, and an apparent negative relationship between creatinine and age only in S. apella. CONCLUSIONS: Our results indicate that A. caraya has a faster aging mechanism than S. apella, and the higher relative kidney volume in S. apella is suggestive of high metabolic demands in this species.

Hepatic lesions in captive owl monkeys (Aotus infulatus) with ultrasonographic "starry sky" liver.

BACKGROUND: The "starry sky" (SK) liver is ultrasonographic pattern characterized by multiple hyperechogenic foci in hepatic parenchyma. The study aimed to characterize the microscopic hepatic lesions in captive owl monkeys with SK liver. METHODS: Thirty-seven clinically healthy owl monkeys had their liver scanned and 18 of them had liver biopsy. Animals with SK and peliosis hepatis (PH) were subjected to immunohistochemical and molecular screening for Bartonella spp. RESULTS: SK liver occurred in 59.4% (22/37) of the owl monkeys. Biopsied animals showed steatosis, hydropic degeneration, hemosiderosis, PH, and multifocal granulomatous hepatitis. Two monkeys had SK, granulomatous hepatitis, and PH which were negative for Bartonella spp. CONCLUSIONS: PH and granulomatous hepatitis associated with hepatocellular degenerative lesions may present as hyperechoic nodular liver lesions consisted of SK liver; therefore, concomitant occurrence of two lesions or more contributed to the hepatic SK pattern among owl monkeys and such cases might be clinically monitored.

Histopathological lesions of congenital Zika syndrome in newborn squirrel monkeys.

The absence of an adequate animal model for studies has limited the understanding of congenital Zika syndrome (CZS) in humans during the outbreak in America. In this study, we used squirrel monkeys (Saimiri collinsi), a neotropical primate (which mimics the stages of human pregnancy), as a model of Zika virus (ZIKV) infection. Seven pregnant female squirrel monkeys were experimentally infected at three different gestational stages, and we were able reproduce a broad range of clinical manifestations of ZIKV lesions observed in newborn humans. Histopathological and immunohistochemical analyses of early-infected newborns (2/4) revealed damage to various areas of the brain and ZIKV antigens in the cytoplasm of neurons and glial cells, indicative of CZS. The changes caused by ZIKV infection were intrauterine developmental delay, ventriculomegaly, simplified brain gyri, vascular impairment and neuroprogenitor cell dysfunction. Our data show that the ZIKV infection outcome in squirrel monkeys is similar to that in humans, indicating that this model can be used to help answer questions about the effect of ZIKV infection on neuroembryonic development and the morphological changes induced by CZS.

Ultrasonography-guided oocyte recovery in owl monkeys (aotus azarai infulatus).

The study evaluated an echo-guided oocyte recovery technique in owl monkeys. Twelve females were selected for the transabdominal ovum retrieval technique. This procedure collected twenty-six follicles, of which nine oocytes were recovered, without harm to the animals. The technique is feasible and is a minimally invasive protocol for neotropical primates.

Prenatal disorders and congenital Zika syndrome in squirrel monkeys.

During the Zika virus (ZIKV) outbreak in Brazil (2015-2016), the clinical manifestations associated with its infection were complex and included miscarriage and congenital malformations, not previously described. In this study, we evaluated the prenatal conditions of pregnant female squirrel monkeys (Saimiri collinsi) infected during different gestational thirds (GTs) and assessed all clinical aspects, diagnostic imaging, viremia and the immune response. In our study, 75% of the infected animals in the 1st GT group had significant clinical manifestations, such as miscarriage and prolonged viremia associated with a late immune response. Consequently, their neonates showed fetal neuropathology, such as cerebral hemorrhage, lissencephaly or malformations of the brain grooves, ventriculomegaly, and craniofacial malformations. Thus, our study demonstrated the relevance of pregnant squirrel monkeys as a model for the study of ZIKV infection in neonates due to the broad clinical manifestations presented, including the typical congenital Zika syndrome manifestations described in humans.

Hematological and biochemical parameters of Spix's Saddleback Tamarin (Leontocebus fuscicollis) raised in captivity.

The Spix's Saddleback Tamarin, Leontocebus fuscicollis is widely distributed across the Amazon region, but is endangered. This species is serving an important role in biomedical research in captivity. However, reference values for hematological and biochemical parameters are required for the proper characterization of the species. It was therefore the objective of our research to establish these parameters taking into consideration sex and body mass differences in healthy adult Spix's saddleback tamarins. Collecting 2 mL of blood from each individual, 20 animals were examined (7 males, 13 females), and hematological and biochemical parameters were determined using commercial kits. Of the sixteen variables measured, only red blood cell (RBC), hemoglobin (Hb) and hematocrit (Ht) values were significantly higher in males (7.12 ± 0.98 106/mm, 14.98 ± 1.25 g/dL and 48.71 ± 4.91%, respectively), while red cell distribution width (RDW) was higher in females (14.58 ± 1.89%). Of the biochemical parameters measured, only gamma-glutamyl transferase (GGT) enzyme showed higher activity in females (8.08 ± 4.87 U/L), and a high glucose concentration range was observed (102.0 to 521.0 mg/dL) for both sexes. These parameters established with reference ranges for healthy adults provide a reliable reference source for the interpretation of laboratory housed saddleback tamarin.

Anatomopathological lesions of infection caused by Platynosomum illiciens (Braun, 1901) in Neotropical primates kept in captivity.

This study described the hepatobiliary anatomopathological lesions associated with trematode Platynosomum illiciens parasitism in Neotropical primates kept in captivity. In the evaluated organs, we observed portal fibrosis, biliary epithelial hyperplasia, and inflammatory reaction with a predominance of lymphocytes and plasmocytes, and in some cases infiltration of eosinophils and neutrophils.

Experimental yellow fever virus infection in the squirrel monkey (Saimiri spp.) I: gross anatomical and histopathological findings in organs at necropsy.

BACKGROUND: Non-human primates contribute to the spread of the yellow fever virus (YFV) and the establishment of transmission cycles in endemic areas. OBJECTIVE: To describe the severe histopathological aspects of YFV infection, 10 squirrel monkeys were infected with YFV and blood, brain, liver, kidney, spleen, heart, lung, lymph node and stomach were collected at 1-7, 10, 20 and 30 days post-infection (dpi). METHODS: Histopathological analysis and detection of the genome and viral antigens and neutralising antibodies were performed by RT-PCR, immunohistochemistry and neutralisation test, respectively. FINDINGS: Only one animal died from the experimental infection. The genome and viral antigens were detected in all investigated organs (1-30 dpi) and the neutralising antibodies from seven to 30 dpi. The brain contained perivascular haemorrhage (6 dpi); in the liver, midzonal haemorrhage and lytic necrosis (6 dpi) were observed. The kidney had bleeding in the Bowman's capsule and tubular necrosis (6 dpi). Pyknotic lymphocytes were observed in the spleen (1-20 dpi), the lung had haemorrhage (2-6 dpi), in the endocardium it contained nuclear pyknosis and necrosis (2-3 dpi) and the stomach contained blood in the lumen (6 dpi). MAIN FINDINGS: Squirrel monkeys reliably reproduced the responses observed in human cases of yellow fever and, therefore, constitute an excellent experimental model for studies on the pathophysiology of the disease.

Experimental yellow fever virus infection in the squirrel monkey (Saimiri spp. ) I: gross anatomical and histopathological findings in organs at necropsy

BACKGROUND Non-human primates contribute to the spread of the yellow fever virus (YFV) and the establishment of transmission cycles in endemic areas. OBJECTIVE To describe the severe histopathological aspects of YFV infection, 10 squirrel monkeys were infected with YFV and blood, brain, liver, kidney, spleen, heart, lung, lymph node and stomach were collected at 1-7, 10, 20 and 30 days post-infection (dpi). METHODS Histopathological analysis and detection of the genome and viral antigens and neutralising antibodies were performed by RT-PCR, immunohistochemistry and neutralisation test, respectively. FINDINGS Only one animal died from the experimental infection. The genome and viral antigens were detected in all investigated organs (1-30 dpi) and the neutralising antibodies from seven to 30 dpi. The brain contained perivascular haemorrhage (6 dpi); in the liver, midzonal haemorrhage and lytic necrosis (6 dpi) were observed. The kidney had bleeding in the Bowman's capsule and tubular necrosis (6 dpi). Pyknotic lymphocytes were observed in the spleen (1-20 dpi), the lung had haemorrhage (2-6 dpi), in the endocardium it contained nuclear pyknosis and necrosis (2-3 dpi) and the stomach contained blood in the lumen (6 dpi). MAIN FINDINGS Squirrel monkeys reliably reproduced the responses observed in human cases of yellow fever and, therefore, constitute an excellent experimental model for studies on the pathophysiology of the disease.

Renal biochemistry variables and ultrasonographic imaging in healthy Squirrel monkeys (Saimiri collinsi).

BACKGROUND: The combined use of renal biochemistry and ultrasonographic imaging may improve the correct management of renal disease. Although renal disease is frequently observed in nonhuman primates, renal function markers have not yet been studied in Squirrel monkeys (Saimiri collinsi). OBJECTIVES: The aim of this study was to establish normal renal biochemistry variables and ultrasonographic features in Squirrel monkeys. MATERIAL AND METHODS: Renal biochemistry variables and ultrasonographic images were documented in 29 healthy Squirrel monkeys (15 males and 14 females). Urea, serum creatinine (SCr), and uric acid (UA) concentrations were measured by kinetic assay. Cystatin C (CysC) was analyzed by immunonephelometry. A multiple frequency linear array probe (5-12 MHz) was used for ultrasonographic imaging. The studied indicators of renal function were related to sex, age, and body mass. RESULTS: Serum creatinine was influenced by sex and body mass. Serum concentration of urea, UA, and CysC were not influenced by sex, age, and body mass. Ultrasonographic images provided accurate and comprehensive data for making clinical decisions for Squirrel monkeys. The total renal volume was only influenced by the body mass nested in sex and was positively correlated to body mass. Right renal volume was bigger than the left one. CONCLUSION: Normative standards for the renal evaluation, including biochemistry and ultrasonography, in the Squirrel monkey have been established correlated to age, sex, and body mass.

Evaluation of blood pressure in feline night monkeys (Aotus azarae infulatus) under different restraint protocols.

BACKGROUND: Feline night monkeys are very susceptible to stress when handled, and because of that, the use of anesthetic and physical restraint methods should be carefully evaluated. To ensure a safer anesthesia to the animals the present study aimed to observe the influence of four restraint protocols in the blood pressure of Aotus azarae infulatus. METHODS: Blood pressure was obtained from 10 animals subjected to restraining protocols using tiletamine and zolazepam, isoflurane, ketamine and midazolam, and physical restraint and results were evaluated to determine their effect on this exam. RESULTS: Among the restraint protocols tested, the anesthetic isoflurane promoted lower blood pressure values and no differences were observed between values measured in both arms in all groups. CONCLUSION: The results of our study suggest that all restraint protocols tested significantly alter blood pressure in this species of primates.

Antiviral activity of chloroquine against dengue virus type 2 replication in Aotus monkeys.

Dengue virus (DENV) of the Flaviviridae family is a single positive-stranded RNA virus that is transmitted by Aedes aegypti and Aedes albopictus mosquitoes. The objective of this study was to investigate the use of chloroquine (CLQ) as an antiviral drug against dengue virus in monkeys. To analyze the action of the drug in vivo, nonhuman primates groups (Aotus azarai infulatus) were inoculated with a subcutaneous injection of a virulent strain of DENV-2, treated and untreated CLQ. Blood hematological, viremia, and serum biochemical values were obtained from 16 DENV-2-inoculated, treated and untreated; four received only CLQ and one mock-infected Aotus monkeys. Monkey serum samples (day 0-10 post-inoculation) were assayed by reverse transcription polymerase chain reaction and Cytometric Bead Array for determination of viremia and inflammatory cytokines, respectively. Additionally, body temperature and activity levels were determined. In the present work, CLQ was effective on replication of DENV-2 in Aotus monkeys; a time viremia reduction was observed compared with the controls. The concentration of tumor necrosis factor alpha and interferon gamma in the serum of the animals had a statistically significant reduction in the groups treated with CLQ after infection compared with the controls. A significant decrease in systemic levels of the liver enzyme aspartate aminotransferase (AST) was also observed in the animals treated with CLQ after infection compared with the controls. These results suggest that CLQ interferes in DENV-2 replication in Aotus monkeys.

Goniometric evaluation of both forelimbs and hind limbs in two species of Neotropical monkeys.

BACKGROUND: The aim of this study was to evaluate the goniometric measurements of the forelimb and hind limb joints in two species of Neotropical primates maintained in captivity. METHODS: Eighteen intact monkeys organized into two groups were studied: Group 1-9 brown howlers (Alouatta fusca), and Group 2-9 tufted capuchins (Sapajus apella). RESULTS: Significant differences were observed in the maximum flexion of the shoulder, elbow, carpus, stifle, and tarsus (G1>G2); maximum extension and abduction of the shoulder (G1G2) and adduction (G1G2). The ROM was significantly different in all joints: shoulder (G1G2), hip (G1>G2), stifle (G1

Aggressive keloid-mimicking tumor in Melanosuchus niger in captivity / Queloide agressivo imitando tumor em Melanosuchus niger em cativeiro

The objective of this note is to describe a case of exuberant scarring formation, with keloid characteristics and pseudo-tumoral configuration in a male Black caiman (Melanosuchus niger), with an estimated age of 60 years, belonging to the Zoobotanical Park at the Emílio Goeldi Museum, located in Belém, Pará, Brazil. The alteration appeared on the right posterior limb involving two distal phalanges of the lateral digit and measured 12.4cm at the greatest width. The keloid tissue was surgically removed and samples were processed and analyzed histopathologically, revealing growth made up of fibrous connective tissue with the habitual morphology, which was structurally mature in the more central areas.

O objetivo do presente relato é descrever um caso de formação cicatricial exuberante, característica de queloide e configuração pseudo-tumoral em um jacaré-açu (Melanosuchus niger), macho, com idade estimada de 60 anos, pertencente ao Parque Zoobotânico do Museu Emílio Goeldi, situado em Belém, Pará, Brasil. A alteração manifestou-se no membro posterior direito, envolvendo as duas falanges distais do dedo lateral e mediu 12,4cm na maior largura. A neoformação foi removida cirurgicamente e amostras foram processadas e analisadas por histopatologia, que revelou crescimento constituído por tecido conjuntivo fibroso de morfologia habitual, estando estruturalmente maduro nas áreas mais centrais.

The protective effect of Canova homeopathic medicine in cyclophosphamide-treated non-human primates.

BACKGROUND: Canova activates macrophages and indirectly induces lymphocyte proliferation. Here we evaluated the effects of Canova in cyclophosphamide-treated non-human primates. METHODS: Twelve Cebus apella were evaluated. Four animals were treated with Canova only. Eight animals were treated with two doses of cyclophosphamide (50 mg/kg) and four of these animals received Canova. Body weight, biochemistry and hematologic analyses were performed for 40 days. Micronucleus and comet assays were performed for the evaluation of DNA damage. RESULTS: We observed that cyclophosphamide induced abnormal WBC count in all animals. However, the group treated with cyclophosphamide plus Canova presented a higher leukocyte count than that which received only cyclophosphamide. Cyclophosphamide induced micronucleus and DNA damage in all animals. The frequency of these alterations was significantly lower in the Canova group than in the group without this medicine. CONCLUSIONS: Our results demonstrated that Canova treatment minimizes cyclophosphamide myelotoxicity in C. apella.

Lymphocyte proliferation stimulated by activated Cebus apella macrophages treated with a complex homeopathic immune response modifiers.

INTRODUCTION: Canova is a complex homeopathic medicine that enhances a specific immunologic responses against several exogenous and endogenous conditions. Canova activates macrophages both in vivo and in vitro. AIM AND METHOD: We evaluated the effects of macrophages activated by Canova in vivo and ex vitro in the proliferation of lymphocytes. Canova was used to activate Cebus apella macrophages in vivo or ex vitro with Canova. Lymphocytes were cultured with the macrophage culture medium. The analysis of Canova effects in cultured lymphocytes was performed according to the cell cycle phase using flow cytometry. The Interferon gamma and Interleukin-5 cytokines quantification in these lymphocyte culture media was performed by Enzyme-linked immunosorbent assay (ELISA). RESULTS: We observed that Canova actives macrophages in vivo and ex vitro. The lymphocytes cultured in a supplemented medium with macrophages activated by Canova treatment presented a higher number of proliferation cells than lymphocytes not exposed to macrophages activated by Canova. The Interferon gamma and Interleukin-5 cytokines were only observed in the medium of lymphocytes exposed to macrophages activated by Canova. Thus, Canova has potential as a new adjuvant therapy.

Descrição morfológica dos ramos colaterais viscerais da aorta abdominal do macaco-de-cheiro / Morphological description of the collateral visceral branches from abdominal aorta of squirrel monkey

O Saimiri sciureus é um primata não humano existente na região amazônica e não consta na lista de animais em perigo de extinção, de acordo com o IBAMA. Neste trabalho, sistematizou-se os ramos colaterais viscerais da aorta abdominal de seis animais sendo três machos e três fêmeas, os quais tiveram o sistema circulatório preenchido com látex Neoprene, adicionado de contraste radiográfico. O resultado observado foi que a aorta abdominal emitiu ventralmente, como ramo colateral visceral, a artéria celíaca, que se trifurcou nas artérias gástrica esquerda, hepática e lienal. Os ramos da artéria celíaca promovem a irrigação do estômago, duodeno, fígado, pâncreas e baço. A seguir, a aorta abdominal emitiu a artéria mesentérica cranial, de calibre maior que a artéria celíaca e justaposta caudalmente. A artéria mesentérica cranial emitiu ramos que vascularizaram a parte final do duodeno, pâncreas, jejuno, íleo, ceco, cólon maior e cólon menor. A aorta abdominal emitiu lateralmente as artérias renais direita e esquerda. A artéria adrenal esquerda surgiu como um ramo colateral direto da artéria celíaca e a artéria adrenal direita surgiu da artéria renal direita. A artéria mesentérica caudal foi emitida da superfície ventral da aorta abdominal, logo abaixo das artérias renais. Em L6, a aorta abdominal se bifurcou, dando origem às artérias ilíacas externas direita e esquerda e estas deram origem à artéria ilíaca interna e femoral direita e esquerda. As artérias testiculares ou ováricas tiveram origem na artéria ilíaca interna. A continuidade da aorta abdominal deu origem à artéria sacral mediana e a continuidade desta é chamada de artéria caudal mediana. O estudo da espécie em questão é de suma importância para gerar conhecimentos a respeito dos primatas não humanos existentes em nosso país. Dessa forma, pode-se dizer que o Saimiri sciureus é um importante modelo biológico para o desenvolvimento de pesquisas biomédicas.

The Saimiri sciureus is a non human primate that exists in the amazon region, which is not in the list of endangered animals according to IBAMA. So, this research systemized the collateral branches visceral of the abdominal aorta of these animals. It was used six animals, three males and three females, which had the circulatory system filled with latex Neoprene added of radiographic contrast. The results showed that the abdominal aorta ventrally emitted a collateral visceral branch called celiac artery, which originates three branches: left gastric, hepatic and lienal arteries. The branches of the celiac artery promoted the irrigation of the stomach, duodenum, liver, pancreas and spleen. Then the abdominal aorta emitted the cranial mesenteric artery, which was of a larger size than the celiac artery and was caudally juxtaposed. The cranial mesenteric artery supply branches that irrigate the final portion of the duodenum, pancreas, jejunum, ileum, cecum and colon. Abdominal aorta emitted laterally the right and left kidney arteries. The left adrenal artery as a collateral branch of the celiac artery and the right adrenal artery has been originated in the right renal artery. The mesenteric caudal artery was emitted from the ventral surface of the abdominal aorta, below the kidney arteries. In L6, abdominal aorta has two derivations to origin the right and left external iliac arteries and these had given origin to the internal iliac and femoral artery right and left. The testicular or ovary arteries were emitted in internal iliac arteries. Of the abdominal aorta led to origin the sacral median artery and in this sequence there is the caudal median artery. The study of this specie is very important to generate knowledge about non human primates existing in our country. It can be said that Saimiri sciureus is an important biological model for development of biomedical researches.

Dioctophyma renale (Goeze, 1782) in the abdominal cavity of a capuchin monkey (Cebus apella), Brazil.

This study reports a case of parasitism by Dioctophyma renale (Goeze, 1762) encysted in the abdominal cavity of a capuchin monkey (Cebus apella) coming from the Centro Nacional de Primatas, Brazil. The animal was sent to the Veterinary Clinic sector with an increase in abdominal volume and no occurrence of any other clinical signs. Upon palpation, a movable circular mass with a diameter of approximately 10 cm was found. Urinalysis, complete blood count and serum biochemistry were performed without revealing any alterations. The animal was then submitted to an abdominal ultrasound exam. The cyst was punctured and a surgical removal procedure was performed, revealing a brownish-colored cylindrical structure that was already deteriorated, making it impossible to perform morphological analysis and classification. In the sediment of the liquid found, eggs were encountered that had morphological characteristics compatible with D. renale. The objective of this paper is to report the first case of parasitism by D. renale in C. apella (Linnaeus, 1758).