From imaging to precision: low cost and accurate determination of stereotactic coordinates for brain surgery Sapajus apella using MRI.

The capuchin monkey (Sapajus apella), a New World monkey species, exhibits prominent characteristics that make it an ideal model for neuroscience research. These characteristics include its phylogenetic traits, telencephalization coefficient, anatomical structures and pathways, genetic profile, immune responses, cognitive abilities, and complex behavioral repertoires. Traditionally, methodologies for stereotactic neurosurgery in research models have relied on the use of brain atlases. However, this approach can lead to errors due to the considerable variation in brain size and shape among individual monkeys. To address this issue, we developed a protocol for deriving individual coordinates for each monkey using a straightforward and relatively inexpensive method involving MRI imaging. Our protocol utilizes a specially designed, 3D-printed stereotactic head-holder that is safe to use with an MR magnet, non-invasive placement of fiducial markers, and post-processing with open-source software. This approach enhances MRI data visualization, improves anatomical targeting, and refines the design of neurosurgical experiments. Our technique could also prove beneficial in other areas of neuroscience research that require accurate calculation of stereotaxic coordinates. Furthermore, it could be useful for other nonhuman primate species for which brain atlases are typically unavailable.

Experimental Yellow Fever in Squirrel Monkey: Characterization of Liver In Situ Immune Response.

Non-human primates contribute to the spread of yellow fever virus (YFV) and the establishment of transmission cycles in endemic areas, such as Brazil. This study aims to investigate virological, histopathological and immunohistochemical findings in livers of squirrel monkeys (Saimiri spp.) infected with the YFV. Viremia occurred 1-30 days post infection (dpi) and the virus showed a predilection for the middle zone (Z2). The livers were jaundiced with subcapsular and hemorrhagic multifocal petechiae. Apoptosis, lytic and coagulative necrosis, steatosis and cellular edema were also observed. The immune response was characterized by the expression of S100, CD11b, CD57, CD4 and CD20; endothelial markers; stress and cell death; pro and anti-inflammatory cytokines, as well as Treg (IL-35) and IL-17 throughout the experimental period. Lesions during the severe phase of the disease were associated with excessive production of apoptotic pro-inflammatory cytokines, such as IFN-γ and TNF-α, released by inflammatory response cells (CD4+ and CD8+ T lymphocytes) and associated with high expression of molecules of adhesion in the inflammatory foci observed in Z2. Immunostaining of the local endothelium in vascular cells and the bile duct was intense, suggesting a fundamental role in liver damage and in the pathogenesis of the disease.

Experimental Yellow Fever in the Squirrel Monkey (Saimiri spp.): Hematological, Biochemical, and Immunological Findings.

Between 2016 and 2018, Brazil experienced the largest sylvatic epidemic of yellow fever virus (YFV). Despite to the magnitude and rapid spread of the epidemic, little is known about YFV dispersion. The study evaluated whether the squirrel monkey is a good model for yellow fever (YF) studies. Methods: Ten animals were infected with 1 × 106 PFU/mL of YFV, with one negative control. Blood samples were collected daily during the first 7 days and at 10, 20 and 30 days post infection (dpi) for detection of viral load and cytokines by RT-qPCR; measurements of AST, ALT, urea and creatinine were taken; IgM/IgG antibodies were detected by ELISA, and hemagglutination inhibition and neutralization tests were performed. The animals exhibited fever, flushed appearance, vomiting and petechiae, and one animal died. Viremia was detected between 1 and 10 dpi, and IgM/IgG antibodies appeared between 4 and 30 dpi. The levels of AST, ALT and urea increased. The immune responses were characterized by expression of S100 and CD11b cells; endothelial markers (VCAM-1, ICAM-1 and VLA-4), cell death and stress (Lysozyme and iNOS); and pro-inflammatory cytokines (IL-8, TNF-α, and IFN-γ) and anti-inflammatory cytokines (IL-10 and TGF-ß). The squirrel monkeys showed changes similar to those described in humans with YF, and are a good experimental model for the study of YF.

Developmental sources of conservation and variation in the evolution of the primate eye.

Conserved developmental programs, such as the order of neurogenesis in the mammalian eye, suggest the presence of useful features for evolutionary stability and variability. The owl monkey, Aotus azarae, has developed a fully nocturnal retina in recent evolution. Description and quantification of cell cycle kinetics show that embryonic cytogenesis is extended in Aotus compared with the diurnal New World monkey Cebus apella. Combined with the conserved mammalian pattern of retinal cell specification, this single change in retinal progenitor cell proliferation can produce the multiple alterations of the nocturnal retina, including coordinated reduction in cone and ganglion cell numbers, increase in rod and rod bipolar numbers, and potentially loss of the fovea.

Number and topography of cones, rods and optic nerve axons in New and Old World primates.

To better understand the evolution of spatial and color vision, the number and spatial distributions of cones, rods, and optic nerve axon numbers were assessed in seven New World primates (Cebus apella, Saimiri ustius, Saguinus midas niger, Alouatta caraya, Aotus azarae, Calllithrix jacchus, and Callicebus moloch). The spatial distribution and number of rods and cones was determined from counts of retinal whole mounts. Optic axon number was determined from optic nerve sections by electron microscopy. These data were amassed with existing data on retinal cell number and distribution in Old World primates, and the scaling of relative densities and numbers with respect to retinal area, eye and brain sizes, and foveal specializations were evaluated. Regular scaling of all cell types was observed, with the exceptionally large, rod-enriched retina of the nocturnal owl monkey Aotus azarae, and the unusually high cone density of the fovea of the trichromatic howler monkey Alouatta caraya presenting interesting variations on this basic plan. Over all species, the lawful scaling of rods, cones, and retinal ganglion cell number is hypothesized to result from a conserved sequence of cell generation that defends retinal acuity and sensitivity over a large range of eye sizes.

Scaling of neuron number and volume of the pulvinar complex in New World primates: comparisons with humans, other primates, and mammals.

The lateral posterior nucleus and pulvinar (LP-pulvinar complex) are the principal thalamic nuclei associated with the elaborate development of the dorsal and ventral streams of the parietal cortex in primates. In humans, a novel site of origin for a subpopulation of pulvinar neurons has been observed, the ganglionic eminence of the telencephalon. This additional site of neuron origin has been proposed to contribute to the pulvinar's evolutionary expansion (Letinic and Rakic [2001] Nat Neurosci 4:930-936). Studies of neuron number in the LP-pulvinar complex in gibbon, chimpanzee, and gorilla compared to humans, however, did not show that the human LP-pulvinar was unexpectedly large (Armstrong [1981] Am J Phys Anthropol 55:369-383). Here we enlarge the allometric basis for comparison by determining neuron number in the LP-pulvinar complex of six New World primates (Cebus apella, Saimiri ustius, Saguinus midas niger, Alouatta caraya, Aotus azarae, and Callicebus moloch) as well as measuring LP-pulvinar volume in a further set of 24 species including additional primates, carnivores, and rodents. The volume of the LP-pulvinar complex scaled with positive allometry with respect to brain volume across all species examined. The scaling of the number of neurons in the LP-pulvinar complex was extremely similar in New World primates and anthropoid apes, with the human LP-pulvinar value close to the regression line. Comparison of the relative volumes of the LP-pulvinar in the larger sample confirmed this observation, and further demonstrated that both primates and carnivores showed a "grade shift" in its size compared to rodents, with the pulvinar comprising a greater proportion of total brain volume across the board. Diurnal, nocturnal, or crepuscular niche did not discriminate LP-pulvinar size across taxa.

Leishmania (Leishmania) amazonensis-induced cutaneous leishmaniasis in the primate Cebus apella: a model for vaccine trials.

A primate model of leishmaniasis was developed with the objective of future vaccine testing. Lesion development and immunological parameters were studied upon primary and secondary infections. Seven Cebus apella were injected subcutaneously with 2 x 10(6) Leishmania (Leishmania) amazonensis promastigotes. Erythematous nodules appeared 19-29 days p.i., which disappeared 100 days p.i. Four months later, six of the monkeys were challenged with the same inoculum; three of them developed erythematous nodules after 7 days p.i., with ulcer formation in two of these subjects. The lesions were short-lived and all were cured 40 days post challenge. Anti-Leishmania IgG antibodies were detected and they increased after the challenge infection. Leishmania antigen-induced lymphoproliferation was found 1 month post-primary infection, which coincided with IFN-gamma production and lesion development. It decreased to control levels afterwards, but at the time of the challenge dose, it was significantly above the initial level. After the challenge infection, it first increased then decreased sharply at 40 days post-challenge, coinciding with the healing of the lesion. It increased again to a higher level at 60 days post-challenge. Leishmania (Leishmania) amazonensis-infection in C. apella did not induce complete protection against a secondary infection with a homologous parasite although specific antibody production and lymphoproliferation with IFN-gamma production were observed. This fact indicates that vaccine has to be better than infection in the induction of protective immunity, and raises a question on in vitro parameters that should be considered as a counterpart of expected protection induced by vaccine candidate. In addition, we conclude that this is a useful primate model for the evaluation of candidate vaccines.

Chromosome studies in Alouatta belzebul.

The chromosome constitution of Alouatta belzebul was studied with G-, C-, and silver staining. In ten specimens identified as Alouatta belzebul, the chromosome constitution of males (2n = 49) differed from that of females (2n = 50) owing to a Y-autosome translocation. In another single female specimen, probably Alouatta belzebul nigerrima, the diploid chromosome number was also 50, though its karyotype was drastically different from that of Alouatta belzebul belzebul. Chromosome studies, taken together with phenotypic and field observations, suggest that Alouatta belzebul belzebul is phenotypically variable in respect to pelage coloration. This attribute is therefore unreliable for the precise identification of Alouatta belzebul subspecies. Conversely, relatively minor phenotypic differences, allowing for the characterization of subspecies within a same species, coexist with unparalleled, drastic karyotypic divergence. These findings clearly question gross morphological attributes as discriminative characteristics of Alouatta belzebul subspecies.

Scaling the primate lateral geniculate nucleus: niche and neurodevelopment in the regulation of magnocellular and parvocellular cell number and nucleus volume.

New stereological assessments of lateral geniculate nucleus (LGN) neuron numbers and volumes in five New World primates (Cebus apella, Saguinus midas niger, Alouatta caraya, Aotus azarae, and Callicebus moloch) and compiled LGN volumes for an additional 26 mammals were analyzed for a better understanding of visual system evolution. Both the magnocellular (M)- and the parvocellular (P)-cell populations scale allometrically with brain volume in primates, P cells with a significantly higher slope such that, for every increase in M neuron number, P neuron numbers more than double (ln scale; y = 0.89x + 2.42R(2) = 0.664). In diurnal primates, the ratio of P to M cells was slightly but significantly higher than in nocturnal primates. For all mammals, including primates, LGN volume was unrelated to nocturnal or diurnal niche but showed marked differences in slope and intercept depending on taxonomic group. The allometric scaling of M and P cells can be related to the order of neurogenesis, with late-generated P cells increasing with positive allometry compared with the earlier-generated M cells. This developmental regularity links relative foveal representation to relative isocortex enlargement, which is also generated late. The small increase in the P/M cell ratio in diurnal primates may result from increased developmental neuron loss in the M-cell population as it competes for limited termination zones in primary visual cortex.

Pelvimetry in Squirrel Monkeys (Saimiri scireus Linnaeus, 1758) / Pelvimetria em macacos-de-cheiro (Saimiri scireus Linnaeus, 1758)

Saimiri sciureus is a neotropical primate widely used in research. However, there are still difficulties regarding their reproduction in vivaria due to the high incidence of dystocia. Dystocia in primates can be caused by cephalopelvic disproportion and in Squirrel Monkeys, pregnancy of large fetuses were reported. This paper describes pelvimetry data of adult females and subadults in specimens of Squirrel Monkeys, from a research colony of Para, Brazil. Pelvic radiographs were obtained in ventrodorsal projections and the following measurements were taken: superior bi-iliac diameter (SBID); inferior bi-iliac diameter (IBID); bi-iliac average diameter (BIAD); right diagonal diameter (RDD); left diagonal diameter (LDD); sacro-pubic diameter (SPD); Based on the obtained diameters, the entrance area of the pelvis (EAP) was also calculated. The average values of the pelvic diameters and EAP in adult females were SBID 1.714cm, BIAD 1.957cm, IBID 1.686cm, RDD 2.771cm, LDD 2.764cm, SPD 2.543cm and EAP 3.9056cm2; and subadult females: 1.588cm SBID, 1.850cm BIAD, 1.625cm IBID, 2.50cm RDD, LDD 2.474cm, 1.95cm SPD and 2.8293 cm2 EAP. Saimiri sciureus pelvis is characterized as dolichopelvic. There was statistical significance between the values for adult females and subadults to SBID, BIAD, RDD, LDD, SPD and EAP. The values of SBID and IBID were lower when compared to the published data for the same species. The result found on this paper will serve as a basis for future studies using pelvic measurements and dystocia prediction of neotropical primates and comparison between different vivaria.(AU)

Saimiri sciureus é uma espécie de primata neotropical muito utilizada como animal de pesquisa. No entanto ainda há dificuldades em biotérios quanto a sua reprodução devido à alta ocorrência de distocia. A distocia em primatas pode ter origem devido à desproporção cefalopélvica, sendo que em macacos-de-cheiro é relatada a gestação de fetos grandes. O presente trabalho descreve dados de pelvimetria em espécimes de macaco-de-cheiro, fêmeas adultas e subadultas provenientes de uma colônia de pesquisa do Pará, Brasil. Foram realizadas radiografias da pelve em projeção ventrodorsal e por meio destas mensurados os diâmetros biilíaco superior (DBIS); diâmetro biíliaco inferior (DBII); diâmetro biilíaco médio (DBIM); diâmetro diagonal direito (DDD); diâmetro diagonal esquerdo (DDE); diâmetro sacro-púbico (DSP); com base nos diâmetros obtidos também foi calculada a área de entrada da pelve (AEP). Os valores médios dos diâmetros pélvicos e da AEP em fêmeas adultas foram: DBIS 1,714cm, DBIM 1,957cm, DBII 1,686cm, DDD 2,771cm, DDE 2,764cm, DSP 2,543cm e AEP 3,9056cm2; e para fêmeas subadultas: 1,588cm DBIS, 1,850cm DBIM, 1,625cm DBII, 2,50cm DDD, 2,474cm DDE, 1,95cm DSP e 2,8293 cm2 AEP. Observou-se que a pelve de Saimiri sciureus é dolicopélvica. Houve diferença estatística significativa entre os valores para fêmeas adultas e subadultas para DBIS, DBIM, DDD, DDE, DSP e AEP. Em comparação com dados da literatura de pelvimetria para S. sciureus observaram-se menores valores de DBIS e DBII. O resultado deste trabalho servirá como base para futuros estudos utilizando-se mensurações pélvicas e predição de distocia em primatas neotropicais e referência para comparação entre S. sciureus de diferentes biotérios.(AU)