Presence of antibody to Cyno-EBV in domestically bred cynomolgus monkeys (Macaca fascicularis).

The presence of an antibody to the Cyno-EBV was studied with Cyno-EBV bearing cell line (Ts-B-9) as antigen in 186 domestically bred cynomolgus monkeys (Macaca fascicularis) ranging in age from birth to 18 yr as well as in 20 adult (older than 10 yr) cynomolgus monkeys of wild origin. The maternal antibody to Cyno-EBV persisted for less than four mo after birth in infant monkeys. After one yr of age, all the domestically bred monkeys were consistently positive for Cyno-EBV. In the monkeys of wild origin also, the anti-Cyno-EBV antibody was detected without exception. Antibody titers in adult monkeys varied from 1:80 to 1:640.     

Spontaneously occurring congenital polycystic kidney in a cynomolgus monkey (Macaca fascicularis)

Congenital polycystic kidney disease was diagnosed at necropsy in a stillborn male cynomolgus monkey (Macaca fascicularis). This case was very similar to infantile polycystic kidney disease in man and the rhesus monkey, except that no increase in number of intrahepatic bile ducts was observed.     

Comparative studies with six extenders for sperm cryopreservation in the cynomolgus monkey (Macaca fascicularis) and rhesus monkey (Macaca mulatta)

Ejaculated spermatozoa from cynomolgus monkeys and rhesus monkeys were frozen in straws with six different extenders (TTE, DM, mDM, LG-DM, G-DM, and TCG) containing glycerol. Sperm motility and head membrane and acrosomal integrity were evaluated after freezing and thawing, and the cryoprotective effects were compared among the extenders and the two species studied. The results showed that sperm motility and motility recovery with the six extenders were comparable for the cynomolgus and rhesus monkeys. There was no significant difference in sperm motility and head membrane integrity among the six extenders in either the cynomolgus or rhesus monkeys (P>0.05). However, a slightly but statistically lower percentage of acrosomal integrity was found with TCG in both species compared to the other extenders (P<0.05). These findings demonstrate that TTE, DM, mDM, LG-DM, G-DM, and TCG are equally suitable extenders for the cryopreservation of spermatozoa from cynomolgus and rhesus monkeys.     

Determining genetic background in captive stocks of cynomolgus macaques (Macaca fascicularis)

Background As in other model organisms, genetic background in the non‐human primates Macaca mulatta and Macaca fascicularis is an experimental variable that affects the response of other study variables. Genetic background in model organisms is manipulated by breeding schemes but is generally pre‐determined by the source population used to found captive stocks. In M. fascicularis three such sources have been distinguished, however, these are not routinely taken into consideration when designing research. Methods We exemplify a mitochondrial DNA (mtDNA)‐based strategy to trace the maternal geographic origins of M. fascicularis animals of unspecified origins. Results Macaca fascicularis of unspecified origins kept at primate research centers carry mtDNA haplotypes representing all three major genetic subdivisions. Conclusions We suggest that the genetic background of study animals could be better specified in the future using an mtDNA‐based approach, which would enable informed selection of study animals and help reduce variation within and among studies.     

Determining acrosomal status of the cynomolgus monkey (Macaca fascicularis) sperm by fluorescence microscopy

The acrosome of Macaca fascicularis sperm cannot be distinguished by conventional light microscopy, so determining whether sperm are acrosome-intact or-reacted is difficult. We describe methods for labeling the acrosomal region of sperm with two different probes: fluoresceinated Pisum sativum agglutinin and anti-sperm antiserum. Acrosome-intact sperm are much more heavily labeled in the acrosomal region than are acrosome-reacted sperm, providing a simple means of differentiating the two types of sperm. The two probes detect similar numbers of acrosome-reacted sperm following treatment with the divalent cation ionophore, A23187.     

Effect of amino acids on cryopreservation of cynomolgus monkey (Macaca fascicularis) sperm

The effects of three amino acids (proline, glutamine, and glycine) added to the freezing medium Tes-Tris-egg yolk (TTE) for cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa were studied. This is the first report on the effects of amino acids on nonhuman primate sperm cryopreservation. The addition of 5mM proline, 10mM glutamine, and 10 or 20mM glycine each significantly improved post-thaw sperm motility and membrane and acrosome integrity compared with the control (TTE alone). However, a significant decrease in motility and membrane/acrosome integrity was observed when amino acid concentrations increased to 60mM for proline and glutamine, and 80 mM for glycine. The results suggest that adding a limited amount of amino acids to the freezing media is beneficial for freezing cynomolgus monkey sperm.     

Immunohistochemical distribution of leucocyte antigens in lymphoid tissues of cynomolgus monkeys (Macaca fascicularis)

Crossreactivity of antibodies to human leucocyte antigens with lymphoid tissues of cynomolgus monkeys was studied by immunohistochemistry and immunoblotting. Of a total of 54 clusters of differentiation (CD) antigens, 39 were expressed essentially with the same immunostaining patterns in the monkey as in human lymphoid tissues. By immunoblotting L26 (CD20) detected a 35 Kd molecule in the monkey lymph node. Our observations indicated that most of the CD antigens are expressed and can be studied in lymphoid tissues of cynomolgus monkeys.     

Social status and reproductive success of male Macaca fascicularis

Dominant males of nonseasonally breeding species should have more opportunity than seasonal breeders to monopolize access to estrous females and thus enhance their reproductive success. They may also use other strategies for maximizing reproductive success such as producing offspring by relatively high ranking females. To test these hypotheses, the paternity of 35 (80%) of 44 Macaca fascicularis offspring, born over a 28-month period, was established using electrophoretically and serologically defined genetic markers. The social ranks of each of the seven potential fathers and 26 potential mothers were determined by recording outcomes of dyadio agonistic interactions. No evidence could be found to support either hypothesis. Further, in view of previously reported significant, positive correlations between male social rank and sexual activity rates in this study group, sexual activity rates do not appear to predict the number of offspring a male sires in this species.     

Effects of caffeine and dbcAMP on zona pellucida penetration by epididymal spermatozoa of cynomolgus monkeys (Macaca fascicularis)

Spermatozoa mature during epididymal transit, acquiring the abilities to swim progressively, fertilize oocytes, and produce viable offspring. In this study, we investigate the capacity of spermatozoa retrieved from the midcorpus and distal cauda regions of the epididymis of the cynomolgus monkey to penetrate homologous zone pellucida. Successful in vitro fertilization by ejaculated macaque sperm is dependent upon the addition of caffeine and dbcAMP. Therefore, the effect of these cyclic nucleotide mediators was also examined in this study. Results of sperm motion analysis indicate no difference in baseline values (without stimulators) for any motion parameter. With the addition of caffeine and dbcAMP, curvilinear velocity significantly increased only for the distal cauda sperm (P = 0.05). Amplitude of the lateral head displacement was significantly increased for distal cauda sperm (P < 0.01); although elevated above baseline, the increase observed after activation by corpus sperm was significantly lower than that achieved by cauda sperm (P < 0.05). The addition of caffeine and dbcAMP was an absolute requirement for zona penetration by both midcorpus and distal cauda sperm. With activation, zona penetration was significantly decreased for corpus sperm compared to cauda sperm (P < 0.001). These results suggest that cynomolgus monkey sperm reaching the midcorpus region of the epididymis have not completed all of the maturational changes requisite for successful fertilization; this immaturity is evidenced by decreased sperm motion and by impedance at the level of zona penetration. © 1996 Wiley-Liss, Inc.     

Luteinizing hormone receptors in ovaries of the cynomolgus monkey (Macaca fascicularis) during the menstrual cycle

Ovarian luteinizing hormone (LH) receptors were characterized using ovarian tissues from 17 cynomolgus monkeys at different phases of the menstrual cycle. Low binding affinity receptors for ¹²⁵I-LH were observed throughout the menstrual cycle. The binding affinity of these receptors for LH (< 12 × 10¹⁰ M^?1) was approximately the same as that of ovarian LH receptors previously reported in human and nonhuman primates. In addition, high-affinity receptors (17?85 × 10¹⁰ M^?1) were also detected at the mid-luteal phase, during which a large functional corpus luteum was present. Thus the high-affinity LH receptors appear with the formation of the corpus luteum and disappear with its regression. Almost no fluctuation of binding capacity was observed throughout the menstrual cycle (32?112 fmol/ mg of ovarian tissue). The high-affinity LH receptor was judged to be derived from the functional corpus luteum.     

The influence of rearing conditions on maternal behavior in cynomolgus macaques (Macaca fascicularis)

We studied the influence of rearing on the adequacy of maternal behavior by comparing 20 harem-reared and 15 peer-reared primiparous cynomolgus monkeys. We used them plus 11 wild-caught females to extend this comparison to multiparous subjects and also to compare primiparae with multiparae. We observed no behavioral effect of rearing and parity. Primiparous maternal adequacy amounted to 93% in peer-reared subjects and to 95% in harem-reared monkeys. We extensively discuss theories concerning rearing effects. Leitmotifs throughout the discussion are survival value of maternal behavior immediately after parturition and predictive value of this behavior for the further course of maternal behavior. Support is accumulated for the hypothesis that certain tactile and visual experiences suffice for the development of adequate maternal behavior.     

Urinary and fecal immunoglobulin A, cortisol and 11-17 dioxoandrostanes, and serum cortisol in metabolic cage housed female cynomolgus monkeys (Macaca fascicularis)

BACKGROUND AND METHODS: Quantitative enzyme-immunoassays of urinary and fecal immunoglobulin A (IgA), cortisol and 11-17-dioxoandrostanes (11,17-DOA), and serum cortisol in eight metabolic-cage-housed female cynomolgus monkeys were performed. The monkeys were divided into two groups, B and NB. Group B animals were blood sampled every 6 hours, whereas Group NB animals were not handled/blood sampled. RESULTS: No differences were recorded between the amounts of feces and urine excreted by the two groups. Group B animals excreted more urinary cortisol than did Group NB animals indicating that restraint-blood sampling resulted in a stress response. Excreted amounts of IgA and 11,17-DOA (urine and feces) did not differ between the groups. CONCLUSIONS: Urinary cortisol was a reliable marker of the stress associated with repeated blood sampling. Declining amounts of excreted urinary cortisol indicated that cynomolgus monkeys acclimated quickly to repeated blood sampling in metabolism cages. Within and between animal variation in amounts of feces voided demonstrated the importance of expressing fecal markers as 'amounts excreted per time unit per kg body weight' rather than just measuring the concentrations in fecal samples.     

Idiopathic chronic diarrhea associated with dysbiosis in a captive cynomolgus macaque (Macaca fascicularis)

Chronic inflammatory enteric diseases occur commonly in humans and animals, especially in captive bred macaques. However, information about the etiology of idiopathic chronic inflammatory diarrhea in cynomolgus monkeys is limited. In this paper, we reported the unusual case of idiopathic chronic diarrhea in a captive cynomolgus monkey based on microbial, imaging, and microbiome examinations.     

Establishment of a B-lymphoblastoid cell line infected with Epstein-Barr-related virus from a cynomolgus monkey (Macaca fascicularis)

A B-lymphoblastoid cell line (Ts-B) was established from lymph nodes of an apparently healthy cynomolgus monkey. The cells were demonstrated to contain Epstein-Barr virus-related antigens. Moreover, herpesvirus particles were found in the cells by electron microscopy. The cell-free culture medium transformed lymphocytes of cynomolgus rhesus monkeys, and of Japanese monkeys.     

A two‐step extraction method to measure fecal steroid hormones in female cynomolgus monkeys (Macaca fascicularis)

We developed a two‐step extraction method for measuring fecal steroid concentrations. In the first step, distilled water was used to extract steroids from fecal samples. In the second step, a mixture of organic solvents (hexane and ether) was used to re‐extract water extracts that had been transferred to a glass tube. A portion of the upper layer of the organic solvents was transferred to separate assay‐tubes for measurement of estradiol (E2) or progesterone (P), and the organic solvents were evaporated in vacuo. After phosphate‐buffered saline was added to each tube, commercially supplied radioimmunoassay (RIA) kits were used to determine the steroids. We demonstrated the advantages and reliability of this method by using it to assay the steroid hormone concentrations in fecal samples and serum samples collected on the same day from female cynomolgus monkeys who showed normal menstrual cycles and from monkeys who had induced hyperfunction of ovarian steroidgenesis. Different fecal samples from each monkey were used to determine the recovery rate of each steroid in water extraction from the fecal samples and the reproductivity of hormone concentrations in the fecal samples. The results demonstrate that this two‐step method is simple and effective for measuring fecal steroids for monitoring the reproductive status of cynomolgus monkeys, without having to collect serum samples. Am. J. Primatol. 48:291–298, 1999. © 1999 Wiley‐Liss, Inc.     

Occurrence of spontaneous diabetes mellitus in a cynomolgus monkey (Macaca fascicularis) and impaired glucose tolerance in its descendants

Spontaneous diabetes mellitus was diagnosed in a cynomolgus monkey. Clinical and pathological features, such as abnormal glucose tolerance, loss of insulin response, or degeneration of pancreatic beta-cells, resemble human noninsulin-dependent diabetes mellitus. Two descendants of the monkey have developed impaired glucose tolerance and insulin response. Genetic factors seem involved in the appearance of carbohydrate intolerance in this family group of monkeys.     

Mobility of surrogate mothers and persistent neophobia in cynomolgus monkeys (Macaca fascicularis)

In search of the relevant difference between mother rearing and surrogate rearing with respect to the emergence of neophobic behavior in macaques, we tested the hypothesis that mobility of surrogate mothers prevents the development of neophobic behavior in cynomolgus macaques. The experimental group consisted of ten surrogate-reared subjects which during their first year got rides on surrogate mothers moving through the cage. The control group consisted of nine subjects reared with immobile surrogate mothers. Between the age of 7 and 27 months the reactions to two different novel objects were tested. First exposure to one object occurred at 6 months of age and to the other object at 20 months. The kind of object and the age of first exposure made no difference. Contrary to our expectations mobility of surrogates did not affect the incidence of neophobia: most subjects in both groups persistently avoided the objects. This finding seems to support Hofer's hypothesis that the mother acts as a regulator of the infant's behavior and physiology.     

Movement characteristics of ejaculated sperm from cynomolgus monkeys (Macaca fascicularis) analyzed by manual and automated computerized image analysis

Sperm motility is an important indicator of male reproductive function. An automated computerized system was used to measure the movement characteristics of cynomolgus monkey sperm. Swimming velocities were in good agreement with data derived from tracking sperm heads manually with a digitizer, but sperm counting by the system was erroneous. In some ejaculates, there were two subpopulations of sperm with different curvilinear velocities, linearities of swim-paths, and lateral movements of the heads.     

Establishment and characterization of Macaca fascicularis lymphoblastoid cell lines.

A panel of cynomolgus macaque lymphoblastoid cell lines (LCL) was established by transforming peripheral blood mononuclear cells (PBMC) with Herpesvirus papio (HVP), and selected lines were examined by flow cytometry. Results indicate that HVP-transformed macaque LCL are phenotypically heterogeneous and resemble human Epstein-Barr virus (EBV)-transformed LCL in the abundant expression of major histocompatibility complex (MHC) class I and class II molecules. At least some lines are of B cell origin.