Navigating the night: spatial cognition, locomotor and ranging behaviour in Nycticebus species

In this thesis I examined the various ways physical, social, cognitive, and evolutionary factors influence ranging behaviour and spacing in Nycticebus species. I explored these topics with the ultimate goal of informing future conservation actions throughout Southeast Asia, especially translocations and in-situ rehabilitation. Each chapter covers topics, comprising ranging and locomotor behaviour, physical ontogeny, spatial cognition, and sociality. I worked at two field sites between 2014 and 2015 in Java and Vietnam, focused on two species of slow lorises (Nycticebus javanicus ,13 ind. and N. pygmaeus, 6 ind.); augmenting this work with data from long-term datasets and data from existing literature. Despite its frequent use, translocation can have a low success rate in Nycticebus species. I found that home range sizes in translocated pygmy slow loris males were larger than those reported both in wild and other translocated pygmy slow lorises, causing subsequent males to leave the release site. As a natural proxy for human-mediated translocations I analysed long-term data on natal dispersal in the Javan slow loris. I found that both males and females disperse as late as two years after birth. Offspring development and dispersal is rooted in the social interactions between family group members. The skills they acquire during this time help them to find and establish a suitable home range; skills like accessing essential food resources. In the Javan slow loris, I looked at exudate feeding to assess their preparedness. While examining the behavioural repertoire used to access tree gums and the ontogenetic development of the postures needed to access them physically, I found that the Javan slow loris acquires rapidly adult-like limb proportions to help employ vertical postures while feeding on gums highlighting their specialization for vertical clinging. The demand on individuals to locate these resources may act as an important selection pressure on the evolution of spatial cognition. I identified the use of spatial cognition and cognitive maps as tools to revisit goal locations. The distribution of stable resources throughout a home range help individuals create a map of 4 their environment. Another factor affecting spacing and ranging is their social interactions. To reconstruct the ancestral state of primate social organisation in primates, I found that among lorisiforms the presumed promiscuous ancestral state in mammals, then evolved into a dispersed family group at the last common ancestor of lorisiforms. In conclusion, these new data bring to light the difficulties in carrying out a successful translocation, a previously overlooked social dynamic within Nycticebus family groups, the importance of limb proportion in development and vertical clinging, how gum producing trees shape nightly movement, and the evolutionary history of lorisiforms social organizations. This information can be used to formulate new conservation strategies and help refine existing efforts in-situ and in captivity. Especially in translocations, which should amend practices to incorporating the complex sociality and use of spatial cognition to support natural behaviours prior to and throughout the release process

Vertical clingers and gougers: rapid acquisition of adult limb proportions facilitates feeding behaviours in young Javan slow lorises (Nycticebus javanicus)

Animals of all ages need to access essential food resources, either on their own or with the assistance of conspecifics. Rapid physical and behavioural development is one strategy to help young animals reach adulthood. Specialized gum-feeding mammals exploit a food type that is relatively difficult to access and digest and must possess the appropriate adaptions to access large vertical substrates, i.e. tree trunks. Unlike other gum feeding mammals, the Javan slow loris (Nycticebus javanicus) lacks physical structures, such as keeled nails or claws, which animals commonly use to secure themselves to large vertical substrates. To understand how slow lorises of all ages exploit gum, we examined their vertical gouging posture, locomotor behaviour, habitat use during feeding, and their morphometric measures across three age classes (adult, sub-adult, juvenile). Using data collected in Cipaganti, Java, Indonesia between April 2012 and April 2016, we found that individuals of N. javanicus rely on their hand, foot, and limb morphology to maintain vertical gouging postures, in place of claws or keeled nails. Locomotor behaviour, position in tree, and tree DBH showed no significant difference across age classes while feeding. Juveniles were indistinguishable from adults and sub-adults in regards to limb proportion indices, lower leg length, hand span and foot span. Some morphometric measures scaled isometrically e.g. arm length, but those highlighted during prolonged vertical postures scaled allometrically e.g. leg, hand, and foot measures. These findings suggest that the rapid behavioural and physical development of key features may act as an ontogenetic adaptation to facilitate access to a stable food resource at a young age. The Javan slow loris exemplifies the complex relationship that exists between an animal’s diet and the specializations that facilitate access to these food resources

Slow lorises (Nycticebus spp.) display evidence of handedness in the wild and in captivity

It has been suggested that strepsirrhines (lemurs, lorises, and galagos) retain the more primitive left hand preference, whilst monkeys and apes more regularly display a right hand preference at the individual-level. We looked to address questions of laterality in the slow loris (Nycticebus spp.) using spontaneous observations of seven wild individuals, unimanual tests in six captive individuals, and photos of 44 individuals in a bilateral posture assessing handedness at the individual- and group-level. During the unimanual reach task, we found at the individual-level, only four slow lorises showed a hand use bias (R: 3, L:1), Handedness index (HI) ranged from -0.57-1.00. In the wild unimanual grasp task we found at the individual-level two individual showed a right-hand bias, the HI ranged from -0.19-0.70. The bilateral venom pose showed a trend toward a right hand dominant grip in those photographed in captivity, but an ambiguous difference in wild individuals. There are many environmental constraints in captivity that wild animals do not face, thus data collected in wild settings are more representative of their natural state. The presence of right-handedness in these species suggest that there is a need to re-evaluate the evolution of handedness in primates

Survey of practitioners handling slow lorises (Primates: Nycticebus): an assessment of the harmful effects of slow loris bites

Slow lorises (Nycticebus spp.) are one of six venomous mammals, and the only known venomous primate. In the wild envenomation occurs mainly during conspecific competition for mates and territory, but may also be used as an application against parasites or for predator defense. Envenomation in humans is documented, with the most extreme accounts detailing near-fatal anaphylactic shock. From September 2016 – August 2017, we received questionnaire responses from 80 wild animal practitioners working with Nycticebus spp. in zoos, rescue centres and in the wild. We identified 54 practitioners who had experience of being bitten or were otherwise affected by slow loris venom, and an additional 26 incomplete entries. No fatalities were reported. Fifteen respondents noted that medical intervention was required, 12 respondents indicated no reaction to being bitten (9 of these indicated they were wearing gloves). Symptoms for those affected included: anaphylactic shock, paraesthesia, haematuria, dyspnoea, extreme pain, infection and general malaise. Impact of slow loris bites ranged from instantaneous to long-persisting complications, and healing time ranged from 1 day to >8 months. Extremities, including hands and arms, were mostly affected from the bites. Six of nine species of slow loris were reported to bite, with N. pygmaeus being the most common in our sample. We make suggestions regarding the use of these highly threatened yet dangerous primates as unsuitable tourist photo props and zoo animal ambassadors. We discuss the medical complications experienced in relation to protein sensitisation, and bacterial pathogenesis. We recommend future work to ascertain the protein content of slow loris venom to aid in enabling mitigation of risks posed

Slow learning of feeding skills in a nocturnal extractive forager

A long developmental period in animals is often needed to learn skills for adult reproduction and survival, including feeding behaviour. The nocturnal Javan slow loris, Nycticebus javanicus, is unusual in that it consumes a specialized diet of difficult to extract resources, as well as disperses up to a year after sexual maturity. Here, we examined the ontogeny of its feeding behaviour to understand whether learning to feed on difficult resources, including by co-feeding, is related to delayed dispersal. We collected feeding and proximity data on developing and adult wild slow lorises at a long-term field site in Cipaganti, West Java from 2012 to 2018. To determine whether acquisition of insects, exudates, nectar and flowers varied by age, we ran logistic generalized additive mixed models. We found that intake of insects and exudates occurred significantly more in the early stages, and feeding on nectar significantly more in the later stages, of development. Co feeding occurred for all food types, with insects showing the most co-feeding events during early development, and co-feeding on exudates remaining high throughout development. Social learning via co-feeding is a potentially important factor in transmission of dietary information from older individuals, including siblings and parents, to young slow lorises. Differences between immature and adult diets levelled off after sexual maturity and before average dispersal. Together these factors suggest that the period required to learn to forage on difficult items could help explain the delayed dispersal patterns seen in mammals with similar foraging strategies

Venom in furs: Facial masks as aposematic signals in a venomous mammal

The function of colouration in animals includes concealment, communication and signaling, such as the use of aposematism as a warning signal. Aposematism is unusual in mammals, and exceptions help us to understand its ecology and evolution. The Javan slow loris is a highly territorial venomous mammal that has a distinctive facial mask and monochromatic vision. To help understand if they use aposematism to advertise their venom to conspecifics or predators with different visual systems, we studied a population in Java, Indonesia. Using ImageJ, we selected colours from the facial masks of 58 individuals, converted RBG colours into monochromatic, dichromatic and trichromatic modes, and created a contrast index. During 290 captures, we recorded venom secretion and aggressiveness. Using Non-metric Multidimensional Scaling and generalised additive models for location, scale and shape, we found that young slow lorises differ significantly from adults, being both more contrasting and more aggressive, with aggressive animals showing fewer wounds. We suggest aposematic facial masks serve multiple purposes in slow lorises based on age. Change in colouration through development may play a role in intraspecific competition, and advertise toxicity or aggressiveness to competitors and/or predators in juveniles. Aposematic signals combined with intraspecific competition may provide clues to new venomous taxa among mammals

Urine Gastrin Releasing Peptide in the First Week Correlates with BPD and Post-Prematurity Respiratory Disease

Rationale: Bronchopulmonary dysplasia (BPD) is associated with post-prematurity respiratory disease (PRD) in survivors of extreme preterm birth. Identifying early biomarkers that correlate with later development of BPD and PRD may provide insights for intervention. In a preterm baboon model, elevated gastrin-releasing peptide (GRP) is associated with BPD, and GRP inhibition mitigates BPD occurrence. Objective: We performed a prospective cohort study to investigate whether urine GRP levels obtained in the first postnatal week were associated with BPD, PRD, and other urinary biomarkers of oxidative stress. Methods: Extremely low gestational age infants (23-28 completed weeks) were enrolled in a US multicenter observational study, The Prematurity and Respiratory Outcomes Program (http://clinicaltrials.gov/ct2/show/NCT01435187). We used multivariable logistic regression to examine the association between urine GRP in the first postnatal week and multiple respiratory outcomes: BPD, defined as supplemental oxygen use at 36 + 0 weeks postmenstrual age, and post-PRD, defined by positive quarterly surveys for increased medical utilization over the first year (PRD score). Results: A total of 109 of 257 (42%) infants had BPD, and 120 of 217 (55%) had PRD. On adjusted analysis, GRP level more than 80 was associated with BPD (adjusted odds ratio [aOR], 1.83; 95% confidence interval [CI], 1.03-3.25) and positive PRD score (aOR, 2.46; 95% CI, 1.35-4.48). Urine GRP levels correlated with duration of NICU ventilatory and oxygen support and with biomarkers of oxidative stress: allantoin and 8-hydroxydeoxyguanosine. Conclusions: Urine GRP in the first postnatal week was associated with concurrent urine biomarkers of oxidative stress and with later diagnoses of BPD and PRD

Early-Onset Neonatal Sepsis 2015 to 2017, the Rise of Escherichia coli, and the Need for Novel Prevention Strategies

Importance: Early-onset sepsis (EOS) remains a potentially fatal newborn condition. Ongoing surveillance is critical to optimize prevention and treatment strategies. Objective: To describe the current incidence, microbiology, morbidity, and mortality of EOS among a cohort of term and preterm infants. Design, setting, and participants: This prospective surveillance study included a cohort of infants born at a gestational age (GA) of at least 22 weeks and birth weight of greater than 400 g from 18 centers of the Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network from April 1, 2015, to March 31, 2017. Data were analyzed from June 14, 2019, to January 28, 2020. Main outcomes and measures: Early-onset sepsis defined by isolation of pathogenic species from blood or cerebrospinal fluid culture within 72 hours of birth and antibiotic treatment for at least 5 days or until death. Results: A total of 235 EOS cases (127 male [54.0%]) were identified among 217 480 newborns (1.08 [95% CI, 0.95-1.23] cases per 1000 live births). Incidence varied significantly by GA and was highest among infants with a GA of 22 to 28 weeks (18.47 [95% CI, 14.57-23.38] cases per 1000). No significant differences in EOS incidence were observed by sex, race, or ethnicity. The most frequent pathogens were Escherichia coli (86 [36.6%]) and group B streptococcus (GBS; 71 [30.2%]). E coli disease primarily occurred among preterm infants (68 of 131 [51.9%]); GBS disease primarily occurred among term infants (54 of 104 [51.9%]), with 24 of 45 GBS cases (53.3%) seen in infants born to mothers with negative GBS screening test results. Intrapartum antibiotics were administered to 162 mothers (68.9%; 110 of 131 [84.0%] preterm and 52 of 104 [50.0%] term), most commonly for suspected chorioamnionitis. Neonatal empirical antibiotic treatment most frequently included ampicillin and gentamicin. All GBS isolates were tested, but only 18 of 81 (22.2%) E coli isolates tested were susceptible to ampicillin; 6 of 77 E coli isolates (7.8%) were resistant to both ampicillin and gentamicin. Nearly all newborns with EOS (220 of 235 [93.6%]) displayed signs of illness within 72 hours of birth. Death occurred in 38 of 131 infected infants with GA of less than 37 weeks (29.0%); no term infants died. Compared with earlier surveillance (2006-2009), the rate of E coli infection increased among very low-birth-weight (401-1500 g) infants (8.68 [95% CI, 6.50-11.60] vs 5.07 [95% CI, 3.93-6.53] per 1000 live births; P = .008). Conclusions and relevance: In this study, EOS incidence and associated mortality disproportionately occurred in preterm infants. Contemporary cases have demonstrated the limitations of current GBS prevention strategies. The increase in E coli infections among very low-birth-weight infants warrants continued study. Ampicillin and gentamicin remained effective antibiotics in most cases, but ongoing surveillance should monitor antibiotic susceptibilities of EOS pathogens

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Impact of Optimized Breastfeeding on the Costs of Necrotizing Enterocolitis in Extremely Low Birthweight Infants

To estimate risk of NEC for ELBW infants as a function of preterm formula and maternal milk (MM) intake and calculate the impact of suboptimal feeding on NEC incidence and costs