Functional inactivation of pulmonary MAIT cells following 5-OP-RU treatment of non-human primates.

Targeting MAIT cells holds promise for the treatment of different diseases and infections. We previously showed that treatment of Mycobacterium tuberculosis infected mice with 5-OP-RU, a major antigen for MAIT cells, expands MAIT cells and enhances bacterial control. Here we treated M. tuberculosis infected rhesus macaques with 5-OP-RU intratracheally but found no clinical or microbiological benefit. In fact, after 5-OP-RU treatment MAIT cells did not expand, but rather upregulated PD-1 and lost the ability to produce multiple cytokines, a phenotype resembling T cell exhaustion. Furthermore, we show that vaccination of uninfected macaques with 5-OP-RU+CpG instillation into the lungs also drives MAIT cell dysfunction, and PD-1 blockade during vaccination partly prevents the loss of MAIT cell function without facilitating their expansion. Thus, in rhesus macaques MAIT cells are prone to the loss of effector functions rather than expansion after TCR stimulation in vivo, representing a significant barrier to therapeutically targeting these cells.

Dysregulation of IL-17/IL-22 Effector Functions in Blood and Gut Mucosal Gamma Delta T Cells Correlates With Increase in Circulating Leaky Gut and Inflammatory Markers During cART-Treated Chronic SIV Infection in Macaques.

HIV-associated inflammation has been implicated in the premature aging and increased risk of age-associated comorbidities in cART-treated individuals. However, the immune mechanisms underlying the chronic inflammatory state of cART-suppressed HIV infection remain unclear. Here, we investigated the role of γδT cells, a group of innate IL-17 producing T lymphocytes, in the development of systemic inflammation and leaky gut phenotype during cART-suppressed SIV infection of macaques. Plasma levels of inflammatory mediators, intestinal epithelial barrier disruption (IEBD) and microbial translocation (MT) biomarkers, and Th1/Th17-type cytokine functions were longitudinally assessed in blood and gut mucosa of SIV-infected, cART-suppressed macaques. Among the various gut mucosal IL-17/IL-22-producing T lymphocyte subsets including Th17, γδT, CD161+ CD8+ T, and MAIT cells, a specific decline in the Vδ2 subset of γδT cells and impaired IL-17/IL-22 production in γδT cells significantly correlated with the subsequent increase in plasma IEBD/MT markers (IFABP, LPS-binding protein, and sCD14) and pro-inflammatory cytokines (IL-6, IL-1ß, IP10, etc.) despite continued viral suppression during long-term cART. Further, the plasma inflammatory cytokine signature during long-term cART was distinct from acute SIV infection and resembled the inflammatory cytokine profile of uninfected aging (inflammaging) macaques. Overall, our data suggest that during cART-suppressed chronic SIV infection, dysregulation of IL-17/IL-22 cytokine effector functions and decline of Vδ2 γδT cell subsets may contribute to gut epithelial barrier disruption and development of a distinct plasma inflammatory signature characteristic of inflammaging. Our results advance the current understanding of the impact of chronic HIV/SIV infection on γδT cell functions and demonstrate that in the setting of long-term cART, the loss of epithelial barrier-protective functions of Vδ2 T cells and ensuing IEBD/MT occurs before the hallmark expansion of Vδ1 subsets and skewed Vδ2/Vδ1 ratio. Thus, our work suggests that novel therapeutic approaches toward restoring IL-17/IL-22 cytokine functions of intestinal Vδ2 T cells may be beneficial in preserving gut epithelial barrier function and reducing chronic inflammation in HIV-infected individuals.

Effects of transdermal mirtazapine on hyporexic rhesus and cynomolgus macaques (Macaca mulatta and Macaca fascicularis).

BACKGROUND: Hyporexia and weight loss are important indicators of physical and psychological well-being in macaque colonies. An FDA-approved transdermal formulated Mirtazapine (MTZ) shows effectiveness in managing feline hyporexia. This study sought to determine its effectiveness as an appetite stimulant in macaques. METHODS: Fourteen macaques with idiopathic hyporexia, intractable to conventional management were treated with transdermal MTZ (0.5 mg/kg) topically administered to aural pinnae once daily for 14 days. Qualitative food consumption was monitored daily for 6 months. Body weights were collected prior to treatment, every 2 weeks for the first 6 weeks, 10 weeks, and 6 months post-treatment. RESULTS: Transdermal MTZ significantly reduced the frequency of hyporexia during treatment and monthly for 6 months. No significant increase in weight noted until approximately 6 months post-treatment. CONCLUSIONS: Results from this study indicate that a short course of transdermal MTZ is an effective way to increase food consumption in macaques chronically.

Diagnosis and treatment of endometriosis in a hooded capuchin (Sapajus apella).

Spontaneous endometriosis is common in women, great apes and Old World monkeys, but rare in New World monkeys. We report on the clinical diagnosis and successful combination of surgical and medical treatment in a case of spontaneous endometriosis in a diabetic hooded capuchin. Addressing this painful condition had a positive welfare impact.

Treatment of primate platynosomosis: A word of caution about the use of praziquantel in marmosets.

BACKGROUND: The successive reports of Platynosomum illiciens in Neotropical captive primates have increased interest in platynosomosis; however, its treatment is little known. METHODS: Callithrix penicillata (n = 10) naturally and chronically infected with P. illiciens were treated with praziquantel (25 mg/kg BW, three s.c. doses at 24 hours intervals), and coproparasitological tests performed over 67 days. The proportions of primates with a reduction in fecal egg counts (FEC) or negative results progressively increased after treatment, and at the last fecal tests, marmosets were negative. RESULTS AND CONCLUSIONS: Although all primates tolerated the initial days of study well, 40% (4/10) of them died between the 8th and 16th days after the onset of treatment. Clinical signs and necropsies indicated the occurrence of hepatic involvement, biliary obstruction, and cholangitis. Marmosets with a higher previous FEC were more likely to die after treatment. Use of praziquantel should be considered carefully on a case-by-case basis.

Pterygodermatites nycticebi infections in golden lion tamarins (Leontopithecus rosalia rosalia) and aye-ayes (Daubentonia madagascariensis) from a German zoo.

In a golden lion tamarin (Leontopithecus rosalia rosalia) colony kept indoors in a German zoo, two animals presented a sudden onset of reduced general condition, lethargy, and diarrhea. At animal capture for clinical examination, adult nematode stages were observed after stress-induced defecation. Despite treatment, two golden lion tamarins died in the following 2 days. At necropsy, spirurid stages were found in the lungs and intestine. Additionally, adult Pterygodermatites spp. were identified in histopathological samples of intestine and pancreas, confirming the previous diagnosis. Upon diagnosis, all animals were treated with ivermectin (0.2 mg/kg; SC). Thereafter, the general condition of the golden lion tamarins improved, whereby some of them excreted spirurid nematodes over 3 days. Four weeks after treatment, 20 fecal samples from the colony were examined and proved negative for parasitic stages. Given that common German cockroaches (Blattella germanica) are suitable intermediate hosts of Pterygodermatites nycticebi, 30 specimens were collected from seven different locations around the golden lion tamarins housing. Third-stage larvae of Pterygodermatites spp. were recovered from those cockroaches. Regular anthelmintic treatments, coprological screenings, and controls for intermediate hosts were recommended. More than 2 years later, P. nycticebi infection was diagnosed again histopathologically in an aye-aye (Daubentonia madagascariensis) which suddenly died. Coprological analysis confirmed the presence of spirurid eggs. Due to prosimian primates' cockroach-eating habits and given that total cockroach eradication proved impossible, continuous cockroach control strategies and regular treatments of primates are currently performed to prevent further P. nycticebi infections.

Remdesivir (GS-5734) Is Efficacious in Cynomolgus Macaques Infected With Marburg Virus.

Marburg virus (MARV) is a filovirus with documented human case-fatality rates of up to 90%. Here, we evaluated the therapeutic efficacy of remdesivir (GS-5734) in nonhuman primates experimentally infected with MARV. Beginning 4 or 5 days post inoculation, cynomolgus macaques were treated once daily for 12 days with vehicle, 5 mg/kg remdesivir, or a 10-mg/kg loading dose followed by 5 mg/kg remdesivir. All vehicle-control animals died, whereas 83% of animals receiving a 10-mg/kg loading dose of remdesivir survived, as did 50% of animals receiving a 5-mg/kg remdesivir regimen. Remdesivir-treated animals exhibited improved clinical scores, lower plasma viral RNA, and improved markers of kidney function, liver function, and coagulopathy versus vehicle-control animals. The small molecule remdesivir showed therapeutic efficacy in this Marburg virus disease model with treatment initiation 5 days post inoculation, supporting further assessment of remdesivir for the treatment of Marburg virus disease in humans.

Clinical and Pathologic Features of Spontaneous Klebsiella pneumoniae Infection in 9 Rhesus Macaques (Macaca mulatta).

Klebsiella pneumoniae is a gram-negative bacterium found in the environment and as a commensal in humans and animals. In humans, K. pneumoniae is one of the most serious nosocomial infections encountered currently and is characterized by liver abscesses, pneumonia, and bacteremia resulting in meningoencephalitis and endophthalmitis. K. pneumoniae in veterinary medicine is rarely reported in NHP, and recent literature describing this disease is sparse. In our colony of predominantly outdoor-housed rhesus macaques (Macaca mulatta), K. pneumoniae is cultured infrequently from healthy animals during routine screening and is even rarer in sick animals. This report summarizes the clinical and postmortem findings associated with this pathogen in 9 rhesus macaques and compares these results with the disease outcomes reported for humans. In these cases, K. pneumoniae infection was confirmed through culture or PCR testing or both. In our experience, when this bacterium does cause clinical signs, the disease is rapidly progressive and severe. At necropsy of NHP, the findings are strikingly similar to opportunistic Klebsiella-associated syndromes described in humans and include liver abscesses, meningoencephalitis, and endophthalmitis. In addition, many of the affected macaques had similar risk factors to humans that succumb to disease, thus perhaps indicating that rhesus macaques could be a viable model for investigating these syndromes.

YERSINIA PSEUDOTUBERCULOSIS INFECTIONS IN PRIMATES, ARTIODACTYLS, AND BIRDS WITHIN A ZOOLOGICAL FACILITY IN THE UNITED KINGDOM.

Infection with Yersinia pseudotuberculosis can be difficult to diagnose and treat successfully. Twenty-four cases from the Zoological Society of London (ZSL) London Zoo and ZSL Whipsnade Zoo were identified between 2001 and 2019. Husbandry, medical, and postmortem records for six primates, 10 artiodactyls, and eight birds were reviewed to identify common clinical signs and gross lesions. Most cases occurred during the winter; however, an outbreak in four primates occurred during the summer following a period of stress associated with increased ambient noise and activity. Common clinical signs included lethargy (6/6 primates, 4/10 artiodactyls, 4/8 birds) or death without premonitory signs (3/10 artiodactyls, 4/8 birds). Once clinical signs were observed, disease progressed quickly. Poor condition was common in mammals (6/6 primates, 9/10 artiodactyls), but often went undetected until postmortem examination. Neurological signs occurred in three of six primates. Diarrhea and anorexia were uncommon in all animals. Hepatitis was observed in all groups (4/6 primates, 2/10 artiodactyls, 4/8 birds), mesenteric lymphadenomegaly was common in mammals (4/6 primates, 8/10 artiodactyls), and gastroenteritis was common in artiodactyls (7/10). Erythematous, punctate rashes, which have only been reported with yersiniosis in humans, were present in three of six primates. Bacterial cultures from the liver in primates and birds or enlarged mesenteric lymph nodes in artiodactyls were often diagnostic. All isolates were susceptible to marbofloxacin, oxytetracycline, streptomycin, ceftazidime, amoxicillin clavulanic acid, trimethoprim sulfamethoxazole, azithromycin, and doxycycline, and resistant to clindamycin. Histopathology and Perl's Prussian blue stains were performed on available liver samples (n = 18). Intracellular hemosiderin was present in 17 of 18 cases. Additional research is needed to determine if there is a relationship between hemosiderosis and yersiniosis.

Effects of Pregnancy, Outdoor Access, and Antifungal Medication on Hair Loss in Breeding-age Female Pigtailed Macaques (Macaca nemestrina).

Over 18 mo, adult female pigtailed macaques (Macaca nemestrina) housed at a breeding facility in Arizona were monitored every 6 mo for alopecia. The study period coincided with the movement of a majority of animals from primarily outdoor housing to continuous indoor housing and a corresponding decrease in available space. These changes were made due to the newly recognized prevalence of coccidioidomycosis at this site. The effects of pregnancy status, changes in outdoor access or space, and administration of fluconazole for the treatment of coccidioidomycosis on alopecia were explored. In this group of pigtailed macaques pregnancy did not appear to affect alopecia, in contrast to findings from a closely related species, rhesus macaques. Fluconazole administration increased alopecia in older animals but not in the youngest age group. Conversely, the effects of limited outdoor access or decreased space on increasing alopecia were greatest in the youngest group of animals.

Paromomycin sulfate is an effective treatment for balantidiasis in captive cynomolgus monkeys.

There are few effective antimicrobial agents against Balantidium coli infection. The effect of paromomycin sulfate (PS) against B. coli was confirmed in this study of 596 captive cynomolgus monkeys. In several trials, the minimum dose and duration of oral administration of PS were 25 mg/day for 5 + 5 days, with a 2-day withdrawal interval. To facilitate daily PS administration, pumpkin cakes supplemented with PS were made, which not only resulted in precise effects but also increased the efficiency of preparation and administration of PS by the animal care staff. No cysts or trophozoites were detected at 14 or 16 days after the last treatments. There were no obvious differences in blood and biochemical parameters between before and after administration of PS. These results indicate that PS is effective for elimination of B. coli without hematological side effects. These data could contribute to the control of microbiological pathogens during veterinary care and colony management in primate facilities.

Mucosal T Helper 17 and T Regulatory Cell Homeostasis Correlate with Acute Simian Immunodeficiency Virus Viremia and Responsiveness to Antiretroviral Therapy in Macaques.

Depletion of gut T helper 17 (Th17) cells during HIV infection leads to decreased mucosal integrity and increased disease progression. Conversely, T regulatory (Treg) cells may inhibit antiviral responses or immune activation. In HIV elite controllers, a balanced Th17/Treg ratio is maintained in the blood, suggesting a role for these responses in controlling inflammation and viral replication. HIV-infected individuals exhibit a range in responsiveness to combination antiretroviral therapy (cART). Given the link between the Th17/Treg ratio and HIV disease, we reasoned these responses may play a role in cART responsiveness. In this study, we investigated the relationship between the mucosal Th17/Treg ratio to acute simian immunodeficiency virus (SIV) viremia and the response to cART. Nineteen rhesus macaques were infected with highly pathogenic SIVΔB670 virus and cART was initiated 6 weeks postinfection. Mucosal CD4 T cell subsets were assessed by intracellular cytokine staining in the colon and mesenteric lymph nodes. Higher baseline Th17/Treg ratios corresponded with increased acute SIV viremia. Th17/Treg ratios decreased during acute SIV infection and were not restored during cART, and this corresponded to increased gut immune activation (Ki67+), markers of microbial translocation (sCD14), and T cell exhaustion (TIGIT+). Animals that maintained a more balanced mucosal Th17/Treg ratio at the time of cART initiation exhibited a better virological response to cART and maintained higher peripheral CD4 counts. These results suggest mucosal Th17 and Treg homeostasis influences acute viremia and the response to cART, a result that suggests therapeutic interventions that improve the Th17/Treg ratio before or during cART may improve treatment of HIV.

Molecular confirmation and anthelmintic efficacy assessment against natural trichurid infections in zoo-housed non-human primates.

INTRODUCTION: This study aimed to assess Trichuris species infection and evaluate the anthelmintic efficacy of fenbendazole and ivermectin against natural trichurid infections in non-human primates (NHPs), kept at Mahendra Chaudhury (MC) Zoological Park, Chhatbir, India. MATERIALS AND METHODS: Molecular confirmation of Trichuris infection was carried out using polymerase chain reaction targeting internal transcribed spacer sequences, and anthelmintic efficacy was assessed by fecal egg count reduction test, respectively. RESULTS: A 710 base pair product confirmed Trichuris species infection in NHPs. Fenbendazole, 10 mg/kg body weight orally for 5 consecutive days and ivermectin, 100 µg/kg body weight orally for 3 alternate days proved effective and showed a maximum fecal egg reduction of 99.20% and 100% (P < .05) at day 7 post-treatment. CONCLUSIONS: This study highlighted the molecular confirmation of Trichuris species in non-human primates and its management using fenbendazole and ivermectin.

Critical analysis of a doxycycline treatment trial of rhesus macaques infected with Borrelia burgdorferi.

A critical analysis was conducted of a doxycycline treatment trial of Indian rhesus macaques. In this treatment trial, the investigators attempted to infect the primates with Borrelia burgdorferi sensu stricto by at least 10 tick bites from artificially infected ticks. None of the primates became ill; nevertheless, 5 primates were treated with a 28-day course of oral doxycycline. In contrast to the conclusions of the authors, the data did not convincingly document the existence of viable B. burgdorferi in antibiotic-treated primates. The investigators were unable to cultivate the spirochete from any animal after treatment using highly sensitive in vitro methods. Like many prior animal studies, the current study also did not document that the doxycycline exposure in these animals was similar to that expected in humans. Numerous additional methodologic problems are discussed.

Use of Favipiravir to Treat Lassa Virus Infection in Macaques.

Lassa virus, the cause of Lassa fever in humans, is endemic to West Africa. Treatment of Lassa fever is primarily supportive, although ribavirin has shown limited efficacy if administered early during infection. We tested favipiravir in Lassa virus-viremic macaques and found that 300 mg/kg daily for 2 weeks successfully treated infection.

Management of Ocular Human herpesvirus 1 Infection in a White-faced Saki Monkey (Pithecia pithecia).

A 20-y-old male intact white-faced saki monkey (Pithecia pithecia) presented with an acute ocular disease of the right eye. Clinical signs included periocular swelling, conjunctivitis, and anisocoria with a miotic right pupil. Conjunctival swabs were positive for Human herpesvirus 1 (HHV1) according to PCR amplification with sequencing. Initial clinical signs resolved with supportive treatment, and the animal was managed chronically by using acyclovir (5 mg/kg PO twice daily) during flare-ups. After more than 2 y, the progression of clinical disease led to enucleation of the right eye. At 2 mo after surgery, acute presentation of severe neurologic signs, including ataxia and blindness, resulted in euthanasia. Histopathology, PCR analysis, and sequencing results were consistent with viral encephalitis due to HHV1; coinfection with Pithecia pithecia lymphocryptovirus 1 was identified. This report describes the first case of managed HHV1 infection in a platyrrhine primate and the first case of HHV1 in a white-faced saki monkey that was not rapidly fatal.

SUCCESSFUL TREATMENT OF GENERALIZED DEMODICOSIS IN RED-HANDED TAMARINS ( SAGUINUS MIDAS) USING A SINGLE ADMINISTRATION OF ORAL FLURALANER.

Two adult sibling red-handed tamarins ( Saguinus midas) presented with weight loss and multifocal skin masses. A skin biopsy revealed pyogranulomatous dermatitis with intrafollicular Demodex sp. mites. Subsequent skin scrapes confirmed the presence of live mites within lesions. Initial treatment with topical and oral ivermectin was unsuccessful, and lesions continued to progress. A single dose of fluralaner (Bravecto®, Merck Animal Health, Kenilworth, New Jersey, 07033, USA; 28.125 mg po) was administered to each animal approximately 5 mo after initial presentation. Lesions resolved over the next 3 mo, and all follow-up skin scrapes were negative for both animals. No adverse effects were noted. A single oral administration of fluralaner at 30-35 mg/kg appears adequate and safe for the treatment of generalized demodicosis in red-handed tamarins.

A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset (Callithrix jacchus).

Hearing research has long been facilitated by rodent models, although in some diseases, human symptoms cannot be recapitulated. The common marmoset (Callithrix jacchus) is a small, easy-to-handle New World monkey which has a similar anatomy of the temporal bone, including the middle ear ossicular chains and inner ear to humans, than in comparison with that of rodents. Here, we report a reproducible, safe, and rational surgical approach to the cochlear round window niche for the drug delivery to the inner ear of the common marmoset. We adopted posterior tympanotomy, a procedure used clinically in human surgery, to avoid manipulation of the tympanic membrane that may cause conductive hearing loss. This surgical procedure did not lead to any significant hearing loss. This approach was possible due to the large bulla structure of the common marmoset, although the lateral semicircular canal and vertical portion of the facial nerve should be carefully considered. This surgical method allows us to perform the safe and accurate administration of drugs without hearing loss, which is of great importance in obtaining pre-clinical proof of concept for translational research.

PUSTULAR DERMATITIS CAUSED BY IMPETIGO IN RED-TAILED MONKEYS ( CERCOPITHECUS ASCANIUS).

Impetigo is a bacterial infection of the superficial layer of the epidermis with crusting or bullae caused by Streptococcus spp., Staphylococcus spp., or both. A 14-yr-old red-tailed monkey ( Cercopithecus ascanius) presented with recurrent scabbing and ulceration under the nares over an 8-yr period. Repeated cultures and biopsy samples led to a presumptive diagnosis of impetigo, later confirmed on necropsy. Multiple antibiotic regimens were employed with varying success during multiple episodes, while lesions resolved on their own at other times. This condition has not been previously reported in a nonhuman primate, although it is not uncommon in humans.

Detection of mycobacterial infection in non-human primates using the Xpert MTB/RIF molecular assay.

Tuberculosis is a major public health concern, and diagnostic strategies applied to animal populations are scarce. As part of ongoing efforts to control tuberculosis dissemination at our animal facility, two non-human primates (NHP, Saimiri sciureus) presenting cutaneous lesions were examined for mycobacterial infection. Both animals tested positive for acid-fast bacilli and Mycobacterium tuberculosis using a molecular assay (IS6110 PCR). Animals were euthanized and several samples were tested for M. tuberculosis using the Xpert MTB/RIF assay. Many samples were positive for M. tuberculosis and rifampicin resistance, and some produced mycobacterial growth. Oral swabs from cage mates were then tested with Xpert MTB/RIF, and the majority tested positive for M. tuberculosis and rifampicin resistance, and produced growth in culture. To our knowledge, this is the first report of multidrug-resistant mycobacterial infection in NHP. Additionally, our data shows that the Xpert MTB/RIF assay can be useful as a screening tool for tuberculosis infection in NHP.