Search

Phospholipase C zeta (PLCζ) is a sperm-specific protein that triggers oocyte activation. The analysis of PLCζ expression in human spermatozoa can be used as a diagnostic marker for oocyte activation deficiency. Our laboratory has previously optimized a standard “in-house” assay to determine PLCζ expression in human spermatozoa. However, one study has suggested that an antigen unmasking method (AUM) would be more efficient in visualizing PLCζ in human sperm. This study aimed to compare our established assay and AUM (involving HCl, acidic Tyrode's solution ORIGINAL ARTICLE, and heat). The mean relative fluorescence (RF) intensity of PLCζ in frozen-thawed spermatozoa from fourteen fertile donors stained with the in-house method was significantly higher than three other AUM groups (in-house [mean ± standard error of mean]: 18.87 ± 2.39 arbitrary units [a.u.] vs non-AUM: 11.44 ± 1.61 a.u., AT-AUM: 12.38 ± 1.89 a.u., and HCl-AUM: 12.51 ± 2.16 a.u., P < 0.05, one-way analysis of variance). The mean RF intensity of PLCζ in AT- and HCl-treated spermatozoa from 12 infertile males was not significantly different from that of the non-AUM group. However, the in-house method resulted in the highest RF intensity (12.11 ± 1.36 a.u., P < 0.01). Furthermore, specificity testing of antibody-antigen binding indicated that the in-house method showed more specific binding than spermatozoa treated by the AUM. In conclusion, our in-house method showed superior visualization and reliability than the AUM, thus supporting the continued use of our in-house assay for clinical research screening.

Guanosine triphosphate (GTP) cyclohydrolase I (GCH1) catalyzes the conversion of GTP into dihydroneopterin triphosphate (DHNP). DHNP is the first intermediate of the folate de novo biosynthesis pathway in prokaryotic and lower eukaryotic microorganisms and the tetrahydrobiopterin (BH4) biosynthesis pathway in higher eukaryotes. The de novo folate biosynthesis provides essential cofactors for DNA replication, cell division, and synthesis of key amino acids in rapidly replicating pathogen cells, such as Plasmodium falciparum(P. falciparum), a causative agent of malaria. In eukaryotes, the product of the BH4 biosynthesis pathway is essential for the production of nitric oxide and several neurotransmitter precursors. An increased copy number of the malaria parasite P. falciparum GCH1 gene has been reported to influence antimalarial antifolate drug resistance evolution, whereas mutations in the human GCH1 are associated with neuropathic and inflammatory pain disorders. Thus, GCH1 stands as an important and attractive drug target for developing therapeutics. The GCH1 intrinsic dynamics that modulate its activity remains unclear, and key sites that exert allosteric effects across the structure are yet to be elucidated. This study employed the anisotropic network model to analyze the intrinsic motions of the GCH1 structure alone and in complex with its regulatory partner protein. We showed that the GCH1 tunnel-gating mechanism is regulated by a global shear motion and an outward expansion of the central five-helix bundle. We further identified hotspot residues within sites of structural significance for the GCH1 intrinsic allosteric modulation. The obtained results can provide a solid starting point to design novel antineuropathic treatments for humans and novel antimalarial drugs against the malaria parasite P. falciparum GCH1 enzyme.

The de novo folate synthesis pathway is a well-established drug target in the treatment of many infectious diseases. Antimalarial antifolate drugs have proven to be effective against malaria, however, rapid drug resistance has emerged on the two primary targeted enzymes: dihydrofolate reductase and dihydroptoreate synthase. The need to identify alternative antifolate drugs and novel metabolic targets is of imminent importance. The 6-pyruvol tetrahydropterin synthase (PTPS) enzyme belongs to the tunneling fold protein superfamily which is characterized by a distinct central tunnel/cavity. The enzyme catalyzes the second reaction step of the parasite’s de novo folate synthesis pathway and is responsible for the conversion of 7,8-dihydroneopterin to 6-pyruvoyl-tetrahydropterin. In this study, we examine the structural dynamics of Plasmodium falciparum PTPS using the anisotropic network model, to elucidate the collective motions that drive the function of the enzyme and identify potential sites for allosteric modulation of its binding properties. Based on our modal analysis, we identified key sites in the N-terminal domains and central helices which control the accessibility to the active site. Notably, the N-terminal domains were shown to regulate the open-to-closed transition of the tunnel, via a distinctive wringing motion that deformed the core of the protein. We, further, combined the dynamic analysis with motif discovery which revealed highly conserved motifs that are unique to the Plasmodium species and are located in the N-terminal domains and central helices. This provides essential structural information for the efficient design of drugs such as allosteric modulators that would have high specificity and low toxicity as they do not target the PTPS active site that is highly conserved in humans., Graphical Abstract

The formation of zig-zags, chevrons, Y-junctions and line segments is demonstrated in thin films formed from cylindrical morphology Si-containing rod-coil diblock copolymers and triblock terpolymers under solvent annealing. Directed self-assembly of the block copolymers within trenches yields well-ordered cylindrical microdomains oriented either parallel or transverse to the sidewalls depending on the chemical functionalization of the sidewalls, and the location and structure of concentric bends in the cylinders is determined by the shape of the trenches. The innate etching contrast, the spontaneous sharp bends and junctions, and the range of demonstrated periodicity and line/space ratios make these conformationally asymmetric rod-coil polymers attractive for nanoscale pattern generation.

The assembly of picornavirus capsids proceeds through the stepwise oligomerization of capsid protein subunits and depends on interactions between critical residues known as hotspots. Few studies have described the identification of hotspot residues at the protein subunit interfaces of the picornavirus capsid, some of which could represent novel drug targets. Using a combination of accessible web servers for hotspot prediction, we performed a comprehensive bioinformatic analysis of the hotspot residues at the intraprotomer, interprotomer and interpentamer interfaces of the Theiler&rsquo, s murine encephalomyelitis virus (TMEV) capsid. Significantly, many of the predicted hotspot residues were found to be conserved in representative viruses from different genera, suggesting that the molecular determinants of capsid assembly are conserved across the family. The analysis presented here can be applied to any icosahedral structure and provides a platform for in vitro mutagenesis studies to further investigate the significance of these hotspots in critical stages of the virus life cycle with a view to identify potential targets for antiviral drug design.

Enterovirus 71 can be a severe pathogen in small children and immunocompromised adults. Virus uncoating is a critical step in the infection of the host cell; however, the mechanisms that control this process remain poorly understood. We applied normal mode analysis and perturbation response scanning to several complexes of the virus capsid and present a coarse-graining approach to analyze the full capsid. We show that our method offers an alternative to expressing the system as a set of rigid blocks and accounts for the interconnection between nodes within each subunit and protein interfaces across the capsid. In our coarse-grained approach, the modes associated with capsid expansion are captured in the first three nondegenerate modes and correspond to the changes observed in structural studies of the virus. We show that the resolution of the analysis may be modified without losing information on the global motions leading to uncoating. Perturbation response scanning revealed that a protomer cannot serve as a functional unit to explain deformations of the capsid. Instead, we define a pentamer as the minimum functional unit to investigate changes within the capsid. From the modal analysis and perturbation response scanning, we locate a hotspot region surrounding the fivefold axis. The range of the effect of these single, hotspot residues extend to 140 A. The perturbation of internal capsid residues in this region displayed greatest propensity to capsid expansion, thus indicating the significant role that the RNA genome may play in triggering uncoating.

Enteroviruses are responsible for a multitude of human diseases. Expansion of the virus capsid is associated with a cascade of conformational changes that allow the subsequent release of RNA. For the first time, this study presents a comprehensive bioinformatic screen for the prediction of interacting motifs within intraprotomer interfaces and across respective interfaces surrounding the five-fold and two-fold axes. The results identify a network of conserved motif residues involved in interactions in Enteroviruses that may be critical to capsid stabilisation, providing guidelines towards developing antivirals that interfere with viral expansion during RNA release. This article is protected by copyright. All rights reserved.

Baboons, members of the genus Papio, comprise six closely related species distributed throughout sub-Saharan Africa and southwest Arabia. The species exhibit more ecological flexibility and a wider range of social systems than many other primates. This article summarizes our current knowledge of the natural history of baboons and highlights directions for future research. We suggest that baboons can serve as a valuable model for complex evolutionary processes, such as speciation and hybridization. The evolution of baboons has been heavily shaped by climatic changes and population expansion and fragmentation in the African savanna environment, similar to the processes that acted during human evolution. With accumulating long-term data, and new data from previously understudied species, baboons are ideally suited for investigating the links between sociality, health, longevity and reproductive success. To achieve these aims, we propose a closer integration of studies at the proximate level, including functional genomics, with behavioral and ecological studies.

Objective To investigate the applicability of phospholipase C zeta (PLCζ) analysis in assisting the clinical decision-making process when considering artificial oocyte activation (AOA) for infertile males in assisted reproductive technology. Design Fifty-six males (43 infertile/13 fertile) were screened using our PLCζ assay. Setting Fertility unit/university laboratory. Patient(s) Infertile males with abnormal sperm morphology or total fertilization failure, low fertilization rate ( Intervention(s) We analyzed PLCζ levels in sperm from fertile and infertile males. Eligible patients subsequently underwent intracytoplasmic sperm injection (ICSI)/artificial oocyte activation (AOA) with calcimycin (GM508). Main Outcome Measure(s) PLCζ localization and level and the proportion of sperm expressing PLCζ. Thresholds of PLCζ deficiency, fertilization rates, pregnancy rates, and live birth rates of AOA and non-AOA cycles. Result(s) Compared with 13 fertile controls, 34 of the 43 infertile males had significantly lower levels of PLCζ and/or a significantly lower proportion of sperm exhibiting PLCζ. Of these 34 patients, 15 showed a significant PLCζ reduction in both parameters, which we termed “PLCζ deficiency.” Five PLCζ-deficient patients opted for AOA; all five achieved fertilization, and four achieved clinical pregnancies and live births. The fertilization rate improved significantly from 18.6% (ICSI) to 56.8% (ICSI/AOA). The clinical pregnancy rate and live birth rate with AOA were both 40% per initiated cycle. Youden index analysis revealed that the cutoffs below which infertile males were likely to benefit from AOA were 71% for the proportion of sperm expressing PLCζ and 15.57 arbitrary units for mean PLCζ level. Conclusion(s) PLCζ analysis is a useful diagnostic tool to determine patient eligibility for subsequent AOA treatment.

Baboons are well studied in savannah but less so in more closed habitats. We investigated predation on mammals by olive baboons (Papio anubis) at a geographical and climatic outlier, Gashaka Gumti National Park (Nigeria), the wettest and most forested site so far studied. Despite abundant wildlife, meat eating was rare and selective. Over 16 years, baboons killed 7 bushbuck (Tragelaphus scriptus) and 3 red-flanked duiker (Cephalophus rufilatus), mostly still-lying ‘parked' infants. Taking observation time into account, this is 1 predation per group every 3.3 months - far lower than at other sites. Some features of meat eating resemble those elsewhere; predation is opportunistic, adult males monopolize most prey, a targeted killing bite is lacking and begging or active sharing is absent. Carcass owners employ evasive tactics, as meat is often competed over, but satiated owners may tolerate others taking meat. Other features are unusual; this is only the second study site with predation records for bushbuck and the only one for red-flanked duiker. The atypical prey and rarity of eating mammals probably reflects the difficulty of acquiring prey animals when vegetation cover is dense. Our data support the general prediction of the socioecological model that environments shape behavioural patterns, while acknowledging their intraspecific or intrageneric plasticity.

Objectives Fruit husks are rarely uniformly hard, varying in penetrability via sulci and changes in thickness. We tested whether a hard-food specialist primate i) bites randomly on food fruit husk surfaces to access seeds, or ii) selects areas most easily penetrated by canines. We consider this would occur so as to minimize deployed mechanical force, energetic expenditure and risk of dental breakage when feeding. Methods A sulcus is the natural line of weakness where a dehiscent fruit breaks open. Using fruits dentally opened for seeds by golden-back uacaris (Cacajao ouakary) we: 1) analysed bite mark distribution on surface of four fruits types (hard-with-sulcus, soft-with-sulcus, hard-no-sulcus, soft-no-sulcus); 2) quantified the force needed to penetrate hard and soft fruits at sulci and elsewhere on fruit surface; 3) measured fruit wall thickness and correlated it with bite-mark distribution in all four categories of fruit. Results 1) Bite marks were distributed at random only on surfaces of soft fruits. For other fruits types, bite locations were concentrated at the thinnest areas of husk, either over the entire surface (non-sulcate fruits), or at sulci (sulcate fruits). 2) For hard-husked fruits, areas where uacaris concentrated their bites were significantly easier to penetrate than those where they did not. Conclusions This hard-fruit feeding specialist primate is not biting at random on the surface of diet fruits. To access seeds they are focusing on those areas requiring less force to penetrate. This may be to save energy, to minimize the risk of breaking teeth used in food processing, or a combination of both. The study shows, for the first time, the subtlety by which these powerfully-jawed animals process their diet items. Am J Phys Anthropol 160:113–125, 2016. © 2016 Wiley Periodicals, Inc.

Three popular approaches exist for quantifying personality in animals: behavioral coding in unconstrained and experimental settings and trait assessment. Both behavioral coding in an unconstrained setting and trait assessment aim to identify an overview of personality structure by reducing the behavioral repertoire of a species into broad personality dimensions, whereas experimental assays quantify personality as reactive tendencies to particular stimuli. Criticisms of these methods include that they generate personality dimensions with low levels of cross-study or cross-species comparability (behavioral coding in unconstrained and experimental settings) or that the personality dimensions generated are not ecologically valid, that is, not reflecting naturally occurring behavior (trait assessment and experimental assays). Which method is best for comparative research is currently debated, and there is presently a paucity of personality research conducted in wild subjects. In our study, all three described methods are used to quantify personality in a wild animal subject, the Barbary macaque (Macaca sylvanus). Our results show that the structures generated by unconstrained behavioral coding and trait assessment were not equivalent. Personality dimensions derived from both trait assessments and experimental assays demonstrated low levels of ecological validity, with very limited correlation with behaviors observed in nonmanipulated circumstances. Our results reflect the methodological differences between these quantification methods. Based on these findings and the practical considerations of wild animal research, we suggest future comparative studies of quantification methods within similar methodological frameworks to best identify methods viable for future comparisons of personality structures in wild animals. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

This work was supported by Medical Research Council (MRC, South Africa) and Research Council (RC, Rhodes University) grants. CR and ÖTB thank the National Research Foundation of South Africa (grant number 93690). NU was supported by postgraduate fellowships from the NRF and the German Academic Exchange Service (DAAD) and a Henderson Fellowship from Rhodes University. The early stages of picornavirus capsid assembly and the host factors involved are poorly understood. Since the localisation of viral proteins in infected cells can provide information on their function, antibodies against purified Theiler's murine encephalomyelitis virus (TMEV) GDVII capsids were generated by immunisation of rabbits. The resultant anti-TMEV capsid antibodies recognised a C-terminal region of VP1 but not VP2 or VP3 by Western analysis. Examination of the sites of TMEV capsid assembly by indirect immunofluorescence and confocal microscopy showed that at 5 h post infection, capsid signal was diffusely cytoplasmic with strong perinuclear staining and moved into large punctate structures from 6 to 8 h post infection. A plaque reduction neutralisation assay showed that the anti-TMEV capsid antibodies but not anti-VP1 antibodies could neutralise viral infection in vitro. The VP1 C-terminal residues recognised by the anti-TMEV capsid antibodies were mapped to a loop on the capsid surface near to the putative receptor binding pocket. In silico docking experiments showed that the known TMEV co-receptor, heparan sulfate, interacts with residues of VP1 in the putative receptor binding pocket, residues of VP3 in the adjacent pit and residues of the adjoining VP1 C-terminal loop which is recognised by the anti-TMEV capsid antibodies. These findings suggest that the anti-TMEV capsid antibodies neutralise virus infection by preventing heparan sulfate from binding to the capsid. The antibodies produced in this study are an important tool for further investigating virus-host cell interactions essential to picornavirus assembly. Postprint

Summary MODE-TASK, a novel and versatile software suite, comprises Principal Component Analysis, Multidimensional Scaling, and t-Distributed Stochastic Neighbor Embedding techniques using Molecular Dynamics trajectories. MODE-TASK also includes a Normal Mode Analysis tool based on Anisotropic Network Model so as to provide a variety of ways to analyse and compare large-scale motions of protein complexes for which long MD simulations are prohibitive. Beside the command line function, a GUI has been developed as a PyMOL plugin. Availability and implementation MODE-TASK is open source, and available for download from https://github.com/RUBi-ZA/MODE-TASK. It is implemented in Python and C++. It is compatible with Python 2.x and Python 3.x and can be installed by Conda. Supplementary information Supplementary data are available at Bioinformatics online.

Although plant-inhabiting ants are known to act as effective deterrents to a variety of vertebrate and invertebrate herbivores, this has been reported only once before for primates, a group better known for their predation of ants. In the present study, we investigated the effects that colonies of Pseudomyrmex viduus ants living in individual Macrolobium acaciifolium (Fabaceae) trees have on the rates of visitation and fruit removal by four taxa of seed-predating vertebrates: the primate Cacajao melanocephalus ouakary; macaws (Ara spp.); large parrots (Amazona spp.); and the Northern Amazonian red squirrel (Sciurus igniventris). We found that ant presence significantly reduced both rates of visitation and of fruit removal by C. m. ouakary. The same pattern of reduced fruit removal was also observed for other seed predators (parrots, macaws, and squirrels) but not for visitation rates (although this may be a result of the small sample size). This appears to be only the second-known demonstration of the repellent effect of ants on primates and, indeed, the first for squirrels and psittacine birds. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114, 260–273.

Spatial position within a group affects the value of group-living benefits such as reduced predation risk and improved foraging. The threat of predation, poor nutrition or increased competition from conspecifics can all cause stress. In many species, central positions are known to be more beneficial than peripheral positions in terms of reduced predation, vigilance and foraging. In this study, we examine whether spatial position within a group is associated with stress and anxiety in a troop of olive baboons (Papio anubis). We predicted that the benefits of occupying central positions would be reflected by a reduction in stress and anxiety for animals who spent the most time in the centre of the group. The study subjects appeared to compete actively for the centre of the group. Physiological stress measures (faecal glucocorticoid concentrations) were positively correlated with time spent in central positions. Time spent in central positions was positively correlated with proximity but negatively correlated with vigilance behaviours (alarm barks). Vigilance rates were positively correlated with measures of anxiety (self-scratch frequency). It is suggested that individuals experience chronic stress due to proximity to conspecifics in central positions, whilst perceived predation risk causes anxiety, with perceived predation risk experienced more by individuals on the periphery.

A female mammal's reproductive function and output are limited by the energy she is able to extract from her environment. Previous studies of the interrelationships between energetic circumstances and reproductive function in a variety of mammal species have produced varied results, which do not all support the common assumption that higher female reproductive hormone levels, specifically progesterone, indicate better ovarian function and greater reproductive potential, and are associated with lower energetic stress. In the present study faecal progesterone and glucocorticoid levels were assessed in two troops of olive baboons (Papio anubis) in the same population. They face similar ecological challenges, except that one troop crop-raids, potentially affecting its energetic intake and stress levels. The energy intake of individual females was assessed by combining detailed feeding observations with nutritional analysis of food samples. The crop-raiding troop experienced 50% higher energy intake rates and 50% lower glucocorticoid levels compared to the non-crop-raiding troop alongside substantially lower progesterone levels. This suggests that energetic stress is associated with elevated progesterone levels and may be the cause of the non-crop-raiding troop's lower reproductive output. By comparing groups which differ little, except in terms of food access, and also by directly assessing energy intake, our study addresses some of the design limitations of previous research investigating variation in progesterone levels and energetic stress. It therefore has the potential to contribute to greater understanding of the factors affecting differences in reproductive and stress hormone levels and reproductive function in mammals experiencing different energetic circumstances.

Crop-raiding by primates and bushpigsPotamochoerus porcusis a major cause of human–wildlife conflict around Budongo Forest Reserve, Uganda. In 2006–2007 a project was initiated, with farmer participation, to investigate the efficacy of on-farm techniques to reduce crop-raiding, including guarding and early-warning techniques, fences, plant barriers, trenches, lights and nets. Here, farmers' perceptions of the effectiveness and sustainability of these deterrents were evaluated using semi-structured interviews and direct observations. Factors important to farmers in effective, sustainable and locally appropriate crop-raiding mitigation are that deterrents be cost-effective, easily manipulated, improve guarding efficiency and require minimal labour inputs. Farmers reported paid guards, guard dogs, wire fences, lights and bells/alarms as most effective. This differs from observations that farmers independently maintained certain deterrents that they presumably considered valuable, namely wire fences, guard dogs, bells/alarms, trenches, lights and nets. This evaluation demonstrates the importance of farmers' participation and perceptions in the viability and uptake of crop-raiding deterrents, and the importance of assessing conflict mitigation trials over the long term.

Morphological adaptations related to food processing generally reflect those elements of the diet that represent the greatest biomechanical challenge or that numerically dominate the diet. However, in periods of the annual cycle when the availability of such foods is low, items to which a species has low apparent morphological adaptation may be included in the diet. Here we test the responses of a diet-specialist primate to limitations in the supply of the resource it is specialized to exploit. Uacaris are primarily predators of immature seeds, in seasonally flooded forests in Amazonian Brazil, and have dental specializations to open hard-shelled fruits. We investigated the importance of arthropods in the diet of golden-backed uacaris (Cacajao melanocephalus ouakary), examining their seasonal importance in the uacari diet, and the ways C. m. ouakary used to access them. Using scan and ad libitum sampling of feeding and phenology from botanical study plots to assess fruit availability, we conducted an 18-mo study in Jau National Park, Amazonas State, Brazil. We recorded arthropod predation 298 times, with Cacajao melanocephalus ouakary feeding on 26 invertebrate taxa in ≥11 families and 9 different orders. Uacaris extracted wood-boring beetles dentally from rotting wood and smaller larvae from twigs, stems, and petioles, but this food class did not predominate. This food class (encapsulated foods) constituted 23.4 % of the arthropod records. The majority of arthropod food items were either manually removed from substrates (ants, beetle larvae, caterpillars, fulgorid bugs, grasshoppers, mayflies, spiders, termites, wasps, and a whip-scorpion) or plucked from the air (volant Lepidoptera). Uacaris appeared to avoid toxic caterpillars. Insectivory was most frequent when fruit and seeds were least available. Arthropods seem to be seasonally important to this primate, supplementing or making up for shortfalls in the hard fruits and immature seeds for which uacaris have highly developed dental, and possibly intestinal, adaptations.

Individuals under stress and those of intrinsically low quality may have insufficient resources to invest in developing equally on both sides. Such individuals may thus display higher levels of fluctuating asymmetry (FA)—small, random departures from perfect bilateral symmetry. These small deviations from symmetry are often less than 1 % of trait size, similar to variation in symmetry generated by measurement error. Studies regarding FA must thoroughly assess measurement error if reliable conclusions regarding FA and fitness parameters are to be drawn. Although many studies regarding facial symmetry in humans have been conducted, few have been carried out on non-human primates. The primary aim of this study was to assess whether facial FA in a non-human primate, the olive baboon (Papio anubis) can be reliably measured from digital photographs. Facial FA was measured in three sets of bilaterally symmetrical landmarks from digital images of 35 olive baboons at Gashaka-Gumti National Park, Nigeria. Measurement error was found to be low indicating a high level of reliability based on comparisons of values from two photos of the same individual (r = 0.85). Measurement error was found to be related to the size of the face. This suggests that orientation error increases in large faces and could potentially influence FA scores. These findings highlight the need for FA studies to measure and control error caused by orientation and position of subjects if FA is to be of utility to behavioural ecology and conservation (e.g. if FA reflects whether individuals in a population are suffering from environmental or genetic stress).

Summary Molecular dynamics (MD) determines the physical motions of atoms of a biological macromolecule in a cell-like environment and is an important method in structural bioinformatics. Traditionally, measurements such as root mean square deviation, root mean square fluctuation, radius of gyration, and various energy measures have been used to analyze MD simulations. Here, we present MD-TASK, a novel software suite that employs graph theory techniques, perturbation response scanning, and dynamic cross-correlation to provide unique ways for analyzing MD trajectories. Availability and implementation MD-TASK has been open-sourced and is available for download from https://github.com/RUBi-ZA/MD-TASK, implemented in Python and supported on Linux/Unix.

This work was supported by SIR (Medical Research Council, South Africa) and Research Council (RC, Rhodes University) grants. CR and ÖTB thank the National Research Foundation of South Africa (grant number 93690). NU was supported by postgraduate fellowships from the NRF and the German Academic Exchange Service (DAAD) and a Henderson Fellowship from Rhodes University. The VP1 subunit of the picornavirus capsid is the major antigenic determinant and mediates host cell attachment and virus entry. To investigate the localisation of Theiler's murine encephalomyelitis virus (TMEV) VP1 during infection, a bioinformatics approach was used to predict a surface-exposed, linear epitope region of the protein for subsequent expression and purification. This region, comprising the N-terminal 112 amino acids of the protein, was then used for rabbit immunisation, and the resultant polyclonal antibodies were able to recognise full length VP1 in infected cell lysates by Western blot. Following optimisation, the antibodies were used to investigate the localisation of VP1 in relation to Hsp90 in infected cells by indirect immunofluorescence and confocal microscopy. At 5 hours post infection, VP1 was distributed diffusely in the cytoplasm with strong perinuclear staining but was absent from the nucleus of all cells analysed. Dual-label immunofluorescence using anti-TMEV VP1 and anti-Hsp90 antibodies indicated that the distribution of both proteins colocalised in the cytoplasm and perinuclear region of infected cells. This is the first report describing the localisation of TMEV VP1 in infected cells, and the antibodies produced provide a valuable tool for investigating the poorly understood mechanisms underlying the early steps of picornavirus assembly. Postprint

The Neotropics house two guilds of large arboreal vertebrate seed predators: parrots and the pitheciin primates. Both have diets dominated by immature fruits. The possibility of members of the Pitheciinae (generaCacajao,ChiropotesandPithecia) acting as occasional seed dispersers has been mooted, but not experimentally shown. We combined primate behavioural data and seed germination data from three separate field studies in the Brazilian states of Amazonas and Pará to analyse patterns of post-consumption seed survivorship for seeds discarded by three pitheciin species (Cacajao melanocephalus ouakary,Chiropotes chiropotesandChiropotes albinasus). We then calculated the frequency of dispersal events for four species eaten byC. m. ouakary. All three primate species dropped intact seeds while feeding, and 30.7% of 674 dropped seeds germinated ex situ. Undamaged seeds from unripe and ripe samples germinated (29.3% and 42.7%, respectively), and all three primate species carried some fruits up to 20 m from the parent tree before consuming them. Potential seed-dispersal events varied from 1 (Macrolobium acaciifolium) per fruiting cycle to more than 6500 (Duroia velutina), suggesting that there are differences in dispersal potential. In summary, although they are highly specialized seed predators, these primates may also act as important dispersers for some plant species, and effective dispersal is not restricted to ripe fruits, as immature fruits removed from a tree may continue to mature and the seeds later germinate, a much-neglected aspect of dispersal ecology. The possibility that similar events occur in parrots should be experimentally investigated.

We analyse the behaviour of Cacajao melanocephalus ouakary feeding at patches of germinating seedlings in dried-out flooded forest. Seedlings of Eschweilera tenuifolia (Lecythidaceae) were the most commonly eaten (88.9%). Some seed patches were revisited over several days, while others were consistently ignored. We tested 3 predictions relating uacari terrestrial foraging behaviour to: (1) arboreal escape route proximity, (2) seed patch size choice and (3) temporal patterns of repeat exploitation. Comparison of fed-at and ignored patches revealed significant preferences for larger patches, and for those close to arboreal refuges but distant from dense ground-based vegetation. Support for these predictions is interpreted as evidence for predation risk-sensitive foraging.

Predation of tropical arboreal mammals is rarely observed. Here, we report the first observation of predation of a member of the genus Cacajao, and add to the knowledge of eagle predation in the genus Cebus. We compare responses to predation by Cacajao ouakary and Cebus albifrons with previous studies of congenerics and with other close relatives, and compare the alarm vocalizations of C. ouakary with those of other pitheciines. Reactions to non-predating raptors are reported. Constraints on primate anti-predator responses in flooded and non-flooded habitats are considered.

Sexual selection has driven the evolution and elaboration of a wide variety of displays and ornaments in male nonhuman primates, including capes, cheek flanges, and sexual coloration. Among the most sexually dimorphic of all primates is the drill (Mandrillus leucophaeus), the males of which can be 3 times the mass of females, possess large canines, and exhibit extremely bright sexual skin coloration. However, the function of male coloration in this species has never been examined. Here, we present data on male color (measured objectively using digital photography), dominance rank, measures of male-female association, and key sexual behaviors, of adult male drills (n = 17) living in four semi free-ranging enclosures at the Drill Rehabilitation and Breeding Center in Nigeria. We test the hypothesis that male coloration is a badge of status, indicating dominance rank, and the hypothesis that male coloration attracts females. We found that male coloration did indicate rank, and that high ranking, strongly colored males were more likely to associate with adult females, and more specifically with fully tumescent females. These males also engaged in more sexual activity. However, measures of male-female association and sexual behaviors were not related to male color once rank had been taken into consideration; i.e., for males of a given rank, females did not prefer those that were more colorful. We discuss the results in light of what is known about the wild drill social system, in which unfamiliar individuals may come together in “supergroups,” and in which quality indicators of competitive ability may be particularly important.

Non-invasive methods to quantify components of stress in non-human animals rely typically on the use of physiological or behavioural measures. At the physiological level, stress is usually measured non-invasively in terms of faecal or urinary glucocorticoid output. A common group of behavioural measures used are self-directed behaviours (SDBs), which have been shown to be linked to anxiety, a subset of stress, although a number of authors have explicitly linked SDBs to stress more generally. Whether increased rates of SDBs are likely to be associated with increased faecal glucocorticoid ouput in wild mammals remains unclear. Here, for wild female olive baboons, we show no association between day-to-day changes in levels of SDB and cortisol metabolite excretion. We also show no relationship between long-term mean levels of these variables. We discuss several possible interpretations of our results, including the possibility that SDBs represent a behavioural coping mechanism, helping to ameliorate the physiological stress response.

Baboon sexual swellings are among the largest and most colorful signals displayed by any mammal, and many baboon studies have shown an association between sexual swellings and both female and male sexual behavior. However, the extent to which female behavior and sexual swellings combine to signal the timing of ovulation and the fertile period to males, and the extent to which males use these and other signals when determining patterns of mating behavior, remain key topics of research. Here we assess the social and sexual behavior of both female and male olive baboons with respect to detailed measures of swelling size made from digital photographs, measures of fecal progestogen and estrogen levels, and estimates of the timing of ovulation and the fertile period based on those levels. Female aggression and grooming behavior were unrelated to fecal progestogen and estrogen levels, but there were some significant relationships between these hormonal measures and presenting behaviors. Measures of female behavior collected during the study did not appear to reveal the timing of ovulation or the fertile period. Male consortship behavior was closely tied to fine-scale changes in sexual swelling size, but copulation behavior was not. Copulation behavior of consorting males was, however, linked to the timing of both ovulation and the fertile period, suggesting that males did have knowledge about these timings. Together these results suggest that males used fine-scale swelling size changes when deciding when to consort, but that consorting males did not use fine-scale swelling size changes in deciding when to copulate. We propose that swelling size may advertise the period during which males should consort with females, with other signals available only from closer inspection then used by consorting males to assess the timing of the fertile period more accurately. An important implication of this interpretation is that different males may have access to different signals of ovulation at any one time. Such a system would allow females to offer different males different information simultaneously, perhaps offering a solution to the 'female dilemma' of how females can simultaneously assure and confuse paternity in multi-male societies.

Several authors have suggested that the consumption of plant compounds may have direct effects on wild primate reproductive biology, but no studies have presented physiological evidence of such effects. Here, for two troops of olive baboons (Papio hamadryas anubis) at Gashaka-Gumti National Park, Nigeria, we show major seasonal increases in levels of fecal progesterone metabolites in females, and provide evidence that this is linked to the consumption of natural plant compounds. Increases in fecal progestogen excretion occurred seasonally in all females, in all reproductive states, including lactation. Detailed feeding data on the study animals showed that only one food species is consumed by both troops at the time of observed progestogen peaks, and at no other times of the year: the African black plum, Vitex doniana. Laboratory tests demonstrated the presence of high concentrations of progestogen-like compounds in V. doniana. Together with published findings linking the consumption of a related Vitex species (Vitex agnus castus) to increased progestogen levels in humans, our data suggest that natural consumption of V. doniana was a likely cause of the observed increases in progestogens. Levels of progestogen excretion in the study baboons during periods of V. doniana consumption are higher than those found during pregnancy, and prevent the expression of the sexual swelling, which is associated with ovulatory activity. As consortship and copulatory activity in baboons occur almost exclusively in the presence of a sexual swelling, V. doniana appears to act on cycling females as both a physiological contraceptive (simulating pregnancy in a similar way to some forms of the human contraceptive pill) and a social contraceptive (preventing sexual swelling, thus reducing association and copulation with males). The negative effects of V. doniana on reproduction may be counter-balanced by the wide-range of medicinal properties attributed to plants in this genus. This is the first time that physiological evidence has been presented of direct effects of plant consumption on the reproductive biology of wild primates.

This paper examines crop damage by mammals and the farmers' experiences of crop losses around Gashaka village on the south-western border of Gashaka Gumti National Park, Nigeria. Data were collected via a weekly count of damaged maize crops in 23 fields in the wet season and 30 in the dry season. We examined: (i) seasonal frequency and extent of damage, (ii) losses experienced relative to expected gains, and (iii) aspects of fields in relation to their susceptibility to raiding. Cows caused the greatest amount of damage during the year. Tantalus monkeys were the wild species that caused the most damage. The experience of individual farmers varied from no damage to complete loss of an entire season's crop. Currently the only apparent certainty in predicting losses for farmers was that those farmers with fields closer to wildlife refuges are more likely to experience greater losses. The importance of domestic animal crop damage should not be underestimated in its impact on farmers' livelihoods. Man...