Reference Intervals and Percentiles for Hematologic and Serum Biochemical Values in Captive Bred Rhesus (Macaca mulatta) and Cynomolgus Macaques (Macaca fascicularis)

Simple Summary We explored the effect of age, gender, weight-for-height indices, sedation protocol, and housing conditions on the hematologic and serum biochemical values of captive rhesus and cynomolgus macaques. Several blood parameters demonstrated significant and clinically relevant changes in relation to the investigated variables. The results will provide veterinarians and researchers with important reference intervals for evaluating experimental results and health control from rhesus and cynomolgus macaques. Abstract Several physiological characteristics and housing conditions are known to affect hematologic and serum biochemical values in macaques. However, the studies that have been conducted either report values calculated based on a small number of animals, were designed specifically to document the effect of a particular condition on the normal range of hematologic and serum biochemical values, or used parametric assumptions to calculate hematologic and serum biochemical reference intervals. We conducted a retrospective longitudinal cohort study to estimate reference intervals for hematologic and serum biochemical values in clinically healthy macaques based on observed percentiles without parametric assumptions. Data were obtained as part of the Biomedical Primate Research Centre (Rijswijk, The Netherlands) health monitoring program between 2018 and 2021. In total, 4009 blood samples from 1475 macaques were analyzed with a maximum of one repeat per year per animal. Data were established by species, gender, age, weight-for-height indices, pregnancy, sedation protocol, and housing conditions. Most of the parameters profoundly affected just some hematologic and serum biochemical values. A significant glucose difference was observed between the ketamine and ketamine-medetomidine sedation protocols. The results emphasize the importance of establishing uniform experimental groups with validated animal husbandry and housing conditions to improve the reproducibility of the experiments.


Introduction
Rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis) are frequently used as biomedical research models [1,2]. Monitoring their wellbeing and physical health is mandatory. Hematologic and serum biochemical values are of great importance in evaluating the individual health status of these macaques. However, many factors such as normal growth and maturation, husbandry, gender, relocation, fasting, and anesthesia can affect hematologic and serum biochemical values in macaques . To provide optimal healthcare and to advance our understanding of macaque models of human disease, it is essential to determine the correct reference intervals to avoid inaccurate conclusions from studies on macaques. In addition, macaques are a prey species and therefore tend to mask disease until it is very severe. This can complicate attempts to evaluate the health status or effect of a treatment, vaccination, or therapy on macaques during experimental research. The early detection of underlying disease is especially important when using animal models for human disease to avoid confounding.
Currently, limited variables are investigated for assessing the reference intervals for macaques (mostly age and gender); reference intervals are calculated from relatively small numbers, and parametric assumptions are used . Therefore, hematologic (HER) and serum biochemical (CER) values from a large cohort of domestically bred rhesus and cynomolgus macaques without overt clinical signs of disease were analyzed in this study. As many hematologic and serum biochemical values are not normally distributed, data are presented as observed percentiles without parametric assumptions. We hypothesize that significant differences in HER and CER values in relation to age, gender, weightfor-height indices, used sedation protocol, pregnancy, and housing conditions exist. Our study provides veterinarians and researchers with a complete set of reference intervals for domestically reared macaques.

Materials and Methods
All data were obtained retrospectively from the electronic health and medical records of animals that were housed at the Biomedical Primate Research Centre (BPRC, Rijswijk, Netherlands) between 2018 and 2021. All animals were housed in accordance with Dutch law and international ethical and scientific standards and guidelines (EU Directive 63/2010). All procedures and husbandry were compliant with the above standards and legislations. The BPRC is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC).
The BPRC houses an outbred breeding colony of rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis), consisting of approximately 1000 rhesus and 150 cynomolgus macaques. The colony was formed around 1975 and consisted initially of captive-bred macaques obtained from various accredited suppliers. Later, new breeding lines were introduced on several occasions to maintain the outbred character of the colony.
All animals underwent annual health evaluations that included weight-for-height indices assessment [38] and blood sampling for HER and CER evaluation. In addition, a thorough physical examination including a pregnancy test was performed by a veterinarian.
The animals were not considered to be specific pathogen-free as they were potentially infected with other common subclinical viral pathogens, including simian foamy virus. The colony was negative for Mycobacterium tuberculosis, Salmonella enterica Typhi, and Shigella dysenteriae at the time of sampling.
Qualified animal caretakers observed all animals for injuries, illness, and fecal consistency at least twice daily. Abnormalities were reported to a veterinarian. Parturitions and stillbirths were recorded, and ultrasonography was used to assist in the judgment of the animal being pregnant at the time of blood sampling.

Housing Condition
The majority of the colony was born and bred in our breeding groups with outdoor access. To perform animal experiments, the macaques were relocated to permanent indoor housing. Therefore, all animals that resided in indoor housing all had a history of outdoor access. The specifics of the housing conditions are detailed below.

Outdoor Access
The animals were housed socially in open enclosures holding naturalistic family groups of between 20 and 30 macaques. The enclosures consisted of freely accessible indoor and outdoor enclosures [39]. Both indoor (75 m 2 , 2.85 m high) and outdoor enclosures (208 m 2 , 3.1 m high) consisted of several compartments and visual barriers. Outdoor enclosures were covered by a galvanized wire mesh and had roofed areas, windbreaks, and shaded areas. The floors in the indoor enclosures were provided with wood fiber bedding (Lignocel 3 4 Grade x, J. Rettenmaier & Sohne GmbH + CO, Rosenberg, Germany), whereas outdoor enclosures had sand bedding. The cleaning procedures of the outdoor enclosures consisted of removing feces on a regular basis. As for the indoor enclosures, high-pressure water cleaning, including disinfection (Anistel Surface disinfectant, Tristel Solutions Limited, Cambridgeshire, UK), was performed monthly. Following disinfection, the enclosures were rinsed with clean water, and the floor was wiped dry. After allowing for a 30-40 min air drying period, new bedding was provided. Environmental enrichment consisted of several climbing structures, beams, fire hoses, and sitting platforms. The indoor enclosure's temperature was at minimum 18 • C for rhesus and 21 • C for cynomolgus macaques, with a 12 h light-dark cycle, and with six air changes per hour. Concerning outdoor enclosures, the atmospheric temperature ranged from approximately −5 to 37 • C throughout the year with a maritime climate. The animals were fed commercial monkey pellets (Ssniff, Soest, Germany) supplemented with limited amounts of fruit, vegetables, or grain mixtures in the afternoon. Enrichment-containing food was provided regularly. Municipal water was available ad libitum, provided by automatic water dispensers.
2.1.2. Permanent Indoor Housing but with a History of Outdoor Access (see Section 2.1.1) Pair-housed animals used in various research programs were housed in indoor cages measuring D 2 × W 2 × H 3 m. The room temperature was 20 ± 2 • C, with six air changes per hour, with a 12 h light-dark cycle and relative humidity of 50 ± 10%. The wood fiber bedding of the enclosures was replaced weekly; one week without additional cleaning procedures, one week after rinsing with tap water, and one week after high-pressure water cleaning, including disinfection (Anistel Surface disinfectant). Enrichment such as mirrors and toys were provided weekly. The animals were fed commercial monkey pellets (Ssniff, Soest, Germany), and limited amounts of fruits or vegetables were provided twice daily. All animals also received additional food enrichment items daily. Municipal water was available ad libitum, provided by automatic water dispensers.

Data Collection and Analysis
Blood samples were obtained during annual veterinary examinations between 2018-2021. These annual exams were performed distributed over the year, with an aimed interval of approximately 12 months for each animal. The animals that were included in the analysis did not present clinical signs of diseases based on daily care and observations. The standard health program did not include preventative anti-parasitic treatment. All study animal pedigrees and birth dates were known. Animals were included in the annual veterinary examinations when they were aged 6 and 9 months and over, for rhesus and cynomolgus macaques, respectively, resulting in an age range between 0.5-27.7 and 0.6-26.7 years for rhesus and cynomolgus macaques, respectively. Tables 1 and 2 shows the animal details by the number of repeats by gender and species.
The macaques were fasted overnight (16 h) prior to sample collection while water was maintained available throughout. All blood samples were collected in the morning.   1  120  132  17  25  2  100  91  32  57  3  229  122  49  49  4  263  77  95  17 An effort was made to minimize stress during the capture procedure before sedation. As our institute is well-experienced and progressive in refinement, a continuous training program was developed to establish cooperation during the capture procedure in the grouphoused macaques. During this training program, the animals were trained to voluntarily enter an individual squeeze cage. After an intramuscular injection of the sedative, the body weight was recorded, and blood samples were obtained by qualified caretakers approximately within 20 min. The collection site was sterilized with alcohol 70% and 1 mL EDTA, and 2.5 mL clothed blood samples were collected from a femoral vein by using a 20-gauge Vacuette needle and hub. The samples for HER were collected in EDTA tubes (Greiner Bio-One GmbH, Kremsmünster, Austria) and mixed by inversion, whereas the samples for CER were collected in tubes without anticoagulants (Greiner Bio-One GmbH, Kremsmünster, Austria) and allowed to clot at room temperature for one hour. HER samples with clots or insufficient blood for the anticoagulant were discarded, and hemolytic serum samples were excluded from the CER analysis. Prior to processing and analysis HER samples were once again mixed by inversion and the CER samples were centrifuged at 3000 r.p.m. for 10 minutes. All values were determined on-site at the BPRC within 24 h after sampling.
Both machines were calibrated routinely every 6 months by a service professional.

Investigated Variables
The investigated variables were: • Age: macaques < 4 and ≥4 years of age were compared. The age of 4 years was selected as macaques are considered to reach sexual maturity at the age of 4 years; • Gender: males versus females; • Weight-to-height ratio assessed by weight-for-height indices (WHI). The best measure of relative adiposity and explored the boundaries of overweight and underweight in captive group-housed rhesus and cynomolgus macaques is a species-specific WHI with height to the power of 3.0 (rhesus macaques) and 2.7 (cynomolgus macaques) as it depended least on height and showed high correlation with other relative adiposity measures [38].

Statistics
The reference intervals for a laboratory parameter are defined by the boundaries of the parameter that include 95% of all observations in a population. It is assumed that 95% of a population is normal. The 5% considered not normal are the 2.5% smaller than the lower reference value and the 2.5% greater than the upper reference value. Because most laboratory parameters are not normally distributed, the estimation of the 2.5 and 97.5% percentiles should not be performed when using parametric methods (i.e., Arithmetic Mean ± 1.96 * Standard Deviations). Therefore, the 2.5 and 97.5% percentiles are calculated using nonparametric methods. We used the criteria outlined by Lahti et al. (2002) to decide whether to establish different reference intervals for age and gender groups; the criteria for not partitioning are that >0.9% and <4.1% of the group distributions should be outside the 2.5 and 97.5 percentiles of the common distribution [40]. Nonparametrically estimated reference intervals should be based on group sizes greater than 120; therefore, partitioning resulting in groups with less than 120 observations was not used to define reference intervals [41].
To be able to compare the magnitude of the differences between the groups, betweengroup differences are expressed as the percentual difference of the medians (hereafter referred to as Delta), which is calculated as 100 * (Median Group 1 − Median Group 2)/Mean (Median Group 1 and Median Group 2). Absolute delta values greater than 5% were considered clinically relevant. The statistical significance of between-group differences was evaluated using nonparametric tests (Mann-Whitney for independent and Wilcoxon's signed rank test for paired observations), and p values < 0.001 were considered statistically significant.

Results
Only absolute delta values greater than 5% that were also significant (p < 0.001) were considered clinically relevant and will be discussed below. Tables 3-6 shows the HER and CER intervals and percentiles, excluding glucose, for rhesus and cynomolgus macaques.

Temporal Trends and Sedation Protocol
The most prominent finding was a significant increase in glucose levels between 2018-2019 and 2020-2021 in all ages, genders, and both species (12.2% young females; 13.6% adult females; 10.2% young males; 10.2% adult males in rhesus macaques and 14.9% young females; 15.8% adult females; 15.8% young males; 12.1% adult males in cynomolgus macaques; p < 0.000001) ( Figures S1 and S2). This is a clear indication that the sedation protocol, which changed in 2020 from ketamine to ketamine-medetomidine, influences glucose levels. Therefore, in Tables 3-6, the reference intervals are presented without glucose. The glucose reference intervals are shown in separate tables (Tables 7 and 8). Moreover, higher ALP levels in male rhesus macaques in 2019 compared to 2018 (10.8%; p < 0.000001) and 2020 compared to 2019 (10.5% p < 0.000001) were observed.
We used the criteria outlined by Lahti et al. (2002) to decide whether to establish different reference intervals for age and gender groups; the criteria for not partitioning were that >0.9% and <4.1% of the group distributions should be outside the 2.5 and 97.5 percentiles of the common distribution. Our results showed that reference values for K, Na, and MCHC values can be combined for all age groups and both genders in rhesus macaques (Table 9). Eo can be combined in cynomolgus macaques in all age groups and both genders (Table 9). TBIL can be combined in cynomolgus macaques for both ages but not per gender. In rhesus macaques, Cl, K, PLT, P-LCR, Neut, and Mono can be combined for both ages but not per gender (Table 10). ALT, AST, LDH, Chol, BIC, MCV, MCH, MCHC, PDW, and WBC can be combined for rhesus macaques for gender but not age groups (Table 11).
Concerning glucose, for rhesus macaques, reference intervals can be combined for both genders and both ages, using the reference intervals for sedation protocols A and B (Table 12).

Age versus Gender
Figures S3 and S4 show the percentage difference (Delta %) in the blood values. Animals < 4 years (young animals) and animals > 4 years (adult animals) were compared in both species along with gender.
In both genders of both species, a significant age effect in the ALP levels was observed. Higher ALP levels were observed in young animals compared to adult animals (27.5% in rhesus females, 15.6% in rhesus males, 28.8% in cynomolgus females, and 25.8% in cynomolgus males (all p < 0.000001). Male rhesus adults showed higher ALP levels compared to female rhesus adults (18.9%; p < 0.000001). In young male rhesus macaques, the ALP level was higher compared to young female rhesus macaques (5.1%; p < 0.000001).
Significantly higher ALAT levels were observed in young cynomolgus males compared with adult males (7.2%; p < 0.000001).
AST levels were significantly higher in young rhesus females compared to adult females (6.5%; p < 0.000001). When comparing young males and adult males, higher AST levels were noted in both species (6.8% and 8% in rhesus and cynomolgus macaques, respectively; p < 0.000001).
The TBIL levels in the rhesus macaques were significantly higher in young females compared to adult females (5%; p < 0.000001).
In the rhesus macaques, the Fe levels were higher in adult males compared to young males (6.5%; p < 0.000001) and higher in adult males compared to adult females (5.3%; p < 0.000001).
The Cre levels were higher in adult rhesus compared to young rhesus macaques (7.4% females; 9.1% males; p < 0.000001). In the cynomolgus macaques, we observed higher levels in adult males compared to young males (9.6%; p < 0.000001) and also higher levels in adult males compared to adult females (6.3%; p < 0.000001).
WBC was significantly higher in young rhesus males compared to adult rhesus males (7.9%; p < 0.000001). In both species, a gender effect in adult animals was observed: females showed a higher count than males (7.1% rhesus; 6% cynomolgus; p < 0.000001). Neut was higher in females compared to males in both rhesus (11.9%; p < 0.000001) and cynomolgus macaques (9.3%; p < 0.000001). In rhesus macaques, a higher count in young males compared to adult males (9%; p < 0.000001) was observed. Lymphocytes showed a gender effect with a higher count in males compared to females in both species (6.7% rhesus; 7.2% cynomolgus; p < 0.000001). In addition, higher levels were observed in young animals compared to adults in both genders (12.1% female rhesus; 7.2% male rhesus, 12.5% female cynomolgus; p < 0.000001; 5.3% male cynomolgus (p < 0.001). Eo was higher in young female rhesus macaques compared to adult female rhesus macaques (13.6%; p < 0.00001). In contrast to that, in cynomolgus macaques, a higher count in adult males compared to young males (12.5%; p < 0.00001) was observed. Baso was higher in young female rhesus and young female cynomolgus macaques compared to young males (16.7%; p < 0.0001) and adult females (16.7%; p < 0.000001).
In 2018-2019 (sedation protocol A), we observed higher glucose levels in adult male rhesus macaques compared to adult female adult rhesus macaques (6.7%; p < 0.000001).

Indoor versus Outdoor Housing
Outdoor-housed rhesus macaques had a significantly higher WBC compared to indoorhoused animals (7.6% in females and 5% in males; p < 0.000001) due to an increase in neutrophils (12.5% in females and 7.9% in males p < 0.000001) ( Figures S5 and S6). The TBIL levels in outdoor-housed rhesus macaques were higher compared to indoor-housed rhesus macaques (5.6% in females; p < 0.0001 and 6.5% in males; p < 0.00001).

Discussion
Our study included a uniquely high number of animals, and in addition, all data were calculated using observed percentiles without parametric assumptions, as the majority of hematologic and serum biochemical values do not display a true normal (gaussian) distribution [42]. The results of tests for normality presented in our study confirmed this. When data are not normally distributed, the usual summary statistics (means and standard deviations) and "normal ranges" should be used with caution or should not be used at all. Nevertheless, overall, the obtained reference intervals were comparable with those found in earlier studies [27][28][29][32][33][34][35] and were within the physiological range of our previously accumulated BPRC hematologic and serum biochemical reference intervals.
Nowadays, captive macaques in breeding colonies are maintained socially in outdoor enclosures [39,[43][44][45][46][47]. In the United States of America, already, seventy-five percent of rhesus macaques at national primate research centers are housed in outdoor enclosures [47]. However, this housing type is not always possible for reasons such as climate or research programs (e.g., Animal Biosafety Levels 3 and 4). Animals are then housed indoors, in small groups or pairs. The major benefit of outdoor enclosures is exposure to seasonal fluctuations in light and climate and increased sensory stimulation, which provide great opportunities for exploration and manipulation that all contribute positively to the animals' welfare. However, outdoor housing results in varying background incidences of primary and opportunistic bacterial, viral, parasitic, and fungal pathogens. The soil in the outdoor enclosures can form a host of microbes and parasites. The animals also might have direct or indirect contact with wildlife such as rodents and birds, which can result in exposure to various microorganisms. In addition, indoor facilities are cleaned once per week, but outdoor enclosures are 'cleaned' less frequently. Furthermore, cleaning and disinfection in indoor facilities are more efficient than in outdoor enclosures. Therefore, our finding that animals with outdoor access have higher WBC counts than indoor-housed animals is explainable. Eosinophilia is a central feature of the host response to parasitic infection, but it is generally not observed in protozoal infections. Therefore, it is a more specific sign of helminth infection [48,49]. In helminth infections, the eosinophilia is usually most pronounced early in infection, coinciding with the larval migration through tissues, which then slowly decreases over time. The observed eosinophil levels in our study did not show a clinically relevant difference between indoor and outdoor housing.
Several blood parameters are known to demonstrate significant changes with age in primates [16,50,51]. ALP, as a byproduct of osteoblast activity, is generally higher in children due to rapid bone metabolism and growth. In our study, ALP demonstrated the greatest increase with age, which is in line with the literature, in which it is described that bone growth in young primates produces elevated ALP levels [15,17,18,20,[50][51][52][53].
In non-overweight adult female and male rhesus macaques and female cynomolgus macaques, the ALP showed clinically relevant higher levels compared to overweight animals. Our finding of lower Chol levels in overweight female cynomolgus macaques is similar to Yue et al. (2016) who described lower total Chol levels in captive cynomolgus macaques with overweight compared to lean controls [54]. In addition, in a study with a smaller sample size the Chol levels were described as not being associated with WHI in adult female cynomolgus macaques [55]. In humans, being overweight tends to increase Chol levels in the blood, which is in contrast to our findings in macaques [56].
Physiological changes in pregnancy and puerperium are principally influenced by hormonal changes. Many hematological shifts that occur during these periods are physiological and are of inconsequential concern [57,58]. Our finding that WBC is increased during pregnancy is similar to human data [59]. However, it was observed that a significant increase in the numbers of neutrophils and a decrease in lymphocytes occurs on day 20 of the pregnancy compared to the non-pregnant state in rhesus macaques [57]. Due to the limited number of pregnancies, we did not divide into trimesters.
In macaques, chemical restraint is required to perform reliable physical examinations, diagnostic exercises, and the collection of tissue or body fluid samples (e.g., cerebrospinal fluid). Sedation not only reduces stress on the animal but promotes safety and improves the quality of data collection. Ketamine hydrochloride is the most used drug for the chemical restraint of macaques. However, ketamine sedation is known to cause significant alterations to serum biochemical and hematological variables [14,[23][24][25]30,31,42]. Increased AST, LDH, and CK levels are indicative of local myotoxicity of the injected ketamine, especially when sedations are performed on sequential days. Moreover, a study demonstrated that ketamine sedation reduces leukocyte counts in cynomolgus macaques [31]. Further, reduced leukocyte counts have been observed in rhesus macaques with ketamine sedation alone [23,24]. Ketamine-medetomidine anesthesia is a preferred anesthetic regimen for animals because of its wide safety range and favorable effects on hemodynamics. However, a negative aspect of this regimen is the development of hyperglycemia. α-2 agonists, such as medetomidine, not only impair insulin release from the pancreatic β cells but also increase glucagon release from the α cells. The decrease in the insulin/glucagon ratio results in decreased glucose uptake and increased glucose production by the liver [60]. This is reflected in our results, where we have shown a marked increase (~10%) in glucose levels after medetomidine was introduced to the sedation protocol. This iatrogenic hyperglycemia can hinder the correct interpretation of glucose levels in these animals.
No clinically relevant gender-related differences have been observed for RBC, HGB, and HCT. This is similar to what is described in other reports [3,9,54]. However, others have reported that male macaques show significantly higher RBC, Hb, and HCT values compared to females [11,12,28,32,36,42], similar to observations in humans. Although the general belief is that these results are related to menstrual blood loss in females, scientific justification was not presented.
Climate could be an influential factor in our reference intervals. Wild living primates can face (relative) scarcity and seasonal fluctuations of food resources in their habitat. Macaques adopt active foraging strategies, relying on a variety of food species and adjusting flexibly their food choices based on food availability [61,62]. However, captive macaques are provided a daily balanced diet, resulting in minimal to no variation of intake in nutritional value during the year. Parasite burdens are known to vary seasonally in wildlife, and rainfall is one key aspect of seasonality that has been linked to parasitism in a range of systems. Rainfall can have immediate effects on parasitism rates by affecting parasite survival and movement in the environment, or it can have delayed effects by affecting host susceptibility to parasites through changes in the host body condition or immune function [63][64][65][66][67]. However, the Netherlands has a maritime climate, with no temperature extremes in summer and winter and the absence of a marked wet and dry season (www.weatherbase.com, accessed on 18 January 2023), concluding that it was unlikely that climate was an influential variable for our study.
In the supplementary files, multiple outliers can be observed, the impact of these outliers is minimized because we employed non-parametric statistical methods to establish reference intervals. Hypothetically, this could be subclinical diseased animals as during the annual health evaluation, the veterinarian noted no clinical abnormalities. The animals with clinical abnormalities were excluded from our analysis. No specific follow-up was indicated for those outliers as all animals were observed twice daily by caretakers.
Our updated reference intervals in macaques demonstrate the importance of using appropriate statistical procedures, homogenous animal populations, and establishing uniform and validated animal housing and husbandry conditions to improve the reproducibility of experiments involving blood sampling. It is imperative to have a reliable and comprehensive set of hematologic and serum biochemical reference intervals based on gender, age, weight-for-height indices, pregnancy, sedation protocol, and housing condition to determine the overall health status of an individual macaque or a colony used as a breeding population.

Conclusions
Knowing the effect of age, gender, weight-for-height indices, pregnancy, housing conditions, and sedation protocol on hematologic and serum biochemical values in macaques will help us to distinguish the boundaries between physiological and pathological changes. In addition, this knowledge will advance future research to identify other variables that may affect those parameters.
Supplementary Materials: The following supporting information can be downloaded at: https://www. mdpi.com/article/10.3390/ani13030445/s1, Figure S1: A. Paired comparisons between years in adult rhesus and cynomolgus macaques; numbers indicate percentual differences (Delta) between Year2-Year1. B. Unpaired comparisons between years in adult rhesus and cynomolgus macaques; numbers indicate percentual differences (Delta) between Year2-Year1. C. Unpaired comparisons between years in juvenile rhesus and cynomolgus macaques; numbers indicate percentual differences (Delta) between Year2-Year1. D. Unpaired comparisons between years in rhesus and cynomolgus macaques; numbers indicate percentual differences (Delta) between 20-21-18-19; Figure S2: Laboratory tests according to gender, age, and collection period in rhesus and cynomolgus macaques; Figure S3: Comparisons between gender and age in rhesus and cynomolgus macaques; numbers indicate percentual differences (Delta) between; Figure S4: Laboratory tests according to gender and age in rhesus and cynomolgus macaques; Figure S5: Comparisons between adult rhesus macaques with or without outdoor access, numbers indicate percentual differences (Delta) between outdoor-indoor; Figure S6: Laboratory tests according to outdoor access in adult rhesus and cynomolgus macaques; Figure S7: Comparisons between non-overweight and overweight adult rhesus and cynomolgus; numbers indicate percentual differences (Delta) between non-overweight and overweight; Figure S8: Laboratory tests according to body weight in adult rhesus and cynomolgus macaques; Figure S9: Comparisons between non-pregnant and pregnant adult rhesus and cynomolgus females, numbers indicate percentual differences (Delta) between non-pregnant and pregnant; Figure S10: Laboratory tests according to pregnancy in adult female rhesus and cynomolgus macaques. Institutional Review Board Statement: Ethical review and approval were waived for this study. All data were retrospectively acquired from the annual health evaluations, which are routine procedures as part of the regular health program at the Biomedical Primate Research Center, Rijswijk, the Netherlands.

Informed Consent Statement: Not applicable.
Data Availability Statement: Data are available on reasonable request.