Mating system evolution and worker caste diversity in Pheidole ants.

The efficiency of social groups is generally optimized by a division of labour, achieved through behavioural or morphological diversity of members. In social insects, colonies may increase the morphological diversity of workers by recruiting standing genetic variance for size and shape via multiply mated queens (polyandry) or multiple-breeding queens (polygyny). However, greater worker diversity in multi-lineage species may also have evolved due to mutual worker policing if there is worker reproduction. Such policing reduces the pressure on workers to maintain reproductive morphologies, allowing the evolution of greater developmental plasticity and the maintenance of more genetic variance for worker size and shape in populations. Pheidole ants vary greatly in the diversity of worker castes. Also, their workers lack ovaries and are thus invariably sterile regardless of the queen mating frequency and numbers of queens per colony. This allowed us to perform an across-species study examining the genetic effects of recruiting more patrilines on the developmental diversity of workers in the absence of confounding effects from worker policing. Using highly variable microsatellite markers, we found that the effective mating frequency of the soldier-polymorphic P. rhea (avg. meN = 2.65) was significantly higher than that of the dimorphic P. spadonia (avg. meN = 1.06), despite a significant paternity skew in P. rhea (avg. B = 0.10). Our findings support the idea that mating strategies of queens may co-evolve with selection to increase the diversity of workers. We also detected patriline bias in the production of different worker sizes, which provides direct evidence for a genetic component to worker polymorphism.

Immunogenetic Management Software: a new tool for visualization and analysis of complex immunogenetic datasets.

Here we describe the Immunogenetic Management Software (IMS) system, a novel web-based application that permits multiplexed analysis of complex immunogenetic traits that are necessary for the accurate planning and execution of experiments involving large animal models, including nonhuman primates. IMS is capable of housing complex pedigree relationships, microsatellite-based MHC typing data, as well as MHC pyrosequencing expression analysis of class I alleles. It includes a novel, automated MHC haplotype naming algorithm and has accomplished an innovative visualization protocol that allows users to view multiple familial and MHC haplotype relationships through a single, interactive graphical interface. Detailed DNA and RNA-based data can also be queried and analyzed in a highly accessible fashion, and flexible search capabilities allow experimental choices to be made based on multiple, individualized and expandable immunogenetic factors. This web application is implemented in Java, MySQL, Tomcat, and Apache, with supported browsers including Internet Explorer and Firefox on Windows and Safari on Mac OS. The software is freely available for distribution to noncommercial users by contacting Leslie.kean@emory.edu. A demonstration site for the software is available at http://typing.emory.edu/typing_demo , user name: imsdemo7@gmail.com and password: imsdemo.

Cooperative social clusters are not destroyed by dispersal in a ciliate.

BACKGROUND: The evolution of social cooperation is favored by aggregative behavior to facilitate stable social structure and proximity among kin. High dispersal rates reduce group stability and kin cohesion, so it is generally assumed that there is a fundamental trade-off between cooperation and dispersal. However, empirical tests of this relationship are rare. We tested this assumption experimentally using ten genetically isolated strains of a ciliate, Tetrahymena thermophila. RESULTS: The propensity for social aggregation was greater in strains with reduced cell quality and lower growth performance. While we found a trade-off between costly aggregation and local dispersal in phenotypic analyses, aggregative strains showed a dispersal polymorphism by producing either highly sedentary or long-distance dispersive cells, in contrast to less aggregative strains whose cells were monomorphic local dispersers. CONCLUSION: High dispersal among aggregative strains may not destroy group stability in T. thermophila because the dispersal polymorphism allows social strains to more readily escape kin groups than less aggregative strains, yet still benefit from stable group membership among sedentary morphs. Such dispersal polymorphisms should be common in other social organisms, serving to alter the nature of the negative impact of dispersal on social evolution.

Evolution of dispersal and life history strategies--Tetrahymena ciliates.

BACKGROUND: Considerable attention has focused on how selection on dispersal and other core life-history strategies (reproductive effort, survival ability, colonization capacity) may lead to so-called dispersal syndromes. Studies on genetic variation in these syndromes within species could importantly increase our understanding of their evolution, by revealing whether traits co-vary across genetic lineages in the manner predicted by theoretical models, and by stimulating further hypotheses for experimental testing. Yet such studies remain scarce. Here we studied the ciliated protist Tetrahymena thermophila, a particularly interesting organism due to cells being able to transform into morphs differing dramatically in swim-speed. We investigated dispersal, morphological responses, reproductive performance, and survival in ten different clonal strains. Then, we examined whether life history traits co-varied in the manner classically predicted for ruderal species, examined the investment of different strains into short- and putative long-distance dispersal, while considering also the likely impact of semi-sociality (cell aggregation, secretion of 'growth factors') on dispersal strategies. RESULTS: Very significant among-strain differences were found with regard to dispersal rate, morphological commitment and plasticity, and almost all core life-history traits (e.g. survival, growth performance and strategy), with most of these traits being significantly intercorrelated. Some strains showed high short-distance dispersal rates, high colonization capacity, bigger cell size, elevated growth performance, and good survival abilities. These well performing strains, however, produced fewer fast-swimming dispersal morphs when subjected to environmental degradation than did philopatric strains performing poorly under normal conditions. CONCLUSION: Strong evidence was found for a genetic covariation between dispersal strategies and core life history traits in T. thermophila, with a fair fit of observed trait associations with classic colonizer models. However, the well performing strains with high colonization success and short-distance dispersal likely suffered under a long-distance dispersal disadvantage, due to producing fewer fast-swimming dispersal morphs than did philopatric strains. The smaller cell size at carrying capacity of the latter strains and their poor capacity to colonize as individual cells suggest that they may be adapted to greater levels of dependency on clone-mates (stronger sociality). In summary, differential exposure to selection on competitive and cooperative abilities, in conjunction with selective factors targeting specifically dispersal distance, likely contributed importantly to shaping T. thermophila dispersal and life history evolution.

Molecularly cloned SHIV-1157ipd3N4: a highly replication- competent, mucosally transmissible R5 simian-human immunodeficiency virus encoding HIV clade C Env.

Human immunodeficiency virus type 1 (HIV-1) clade C causes >50% of all HIV infections worldwide, and an estimated 90% of all transmissions occur mucosally with R5 strains. A pathogenic R5 simian-human immunodeficiency virus (SHIV) encoding HIV clade C env is highly desirable to evaluate candidate AIDS vaccines in nonhuman primates. To this end, we generated SHIV-1157i, a molecular clone from a Zambian infant isolate that carries HIV clade C env. SHIV-1157i was adapted by serial passage in five monkeys, three of which developed peripheral CD4(+) T-cell depletion. After the first inoculated monkey developed AIDS at week 137 postinoculation, transfer of its infected blood to a naïve animal induced memory T-cell depletion and thrombocytopenia within 3 months in the recipient. In parallel, genomic DNA from the blood donor was amplified to generate the late proviral clone SHIV-1157ipd3. To increase the replicative capacity of SHIV-1157ipd3, an extra NF-kappaB binding site was engineered into its 3' long terminal repeat, giving rise to SHIV-1157ipd3N4. This virus was exclusively R5 tropic and replicated more potently in rhesus peripheral blood mononuclear cells than SHIV-1157ipd3 in the presence of tumor necrosis factor alpha. Rhesus macaques of Indian and Chinese origin were next inoculated intrarectally with SHIV-1157ipd3N4; this virus replicated vigorously in both sets of monkeys. We conclude that SHIV-1157ipd3N4 is a highly replication-competent, mucosally transmissible R5 SHIV that represents a valuable tool to test candidate AIDS vaccines targeting HIV-1 clade C Env.

The evolution of worker caste diversity in social insects.

Morphological diversification of workers is predicted to improve the division of labor within social insect colonies, yet many species have monomorphic workers. Individual-level selection on the reproductive capacities of workers may counter colony-level selection for diversification, and life-history differences between species (timing of caste determination, colony size, genetic variation available) may mediate the strength of this selection. We tested this through phylogenetically independent contrast analyses on a new data set for 35 ant species. Evidence was found that early divergence of queen-worker developmental pathways may facilitate the evolution of worker diversity because queen-worker dimorphism was strongly positively associated with diversity. By contrast, risks for colonies that invest in specialized workers and colony size effects on costs of worker reproduction seem unlikely to strongly affect the evolution of worker diversity because there was no significant association between colony size and diversity when controlling statistically for queen-worker dimorphism. Finally, worker diversity was greater in species with multiple lineages per colony, and it was negatively associated with relatedness in monogynous species. This could be due to high intracolonial genetic variance favoring the expression and evolution of great worker diversity or to diversity evolving more easily when there is selection for repression of worker reproduction (worker policing).

Control of body size of Lasius niger ant sexuals--worker interests, genes and environment.

In most animals, the survival and reproductive success of males and females is linked to their size. The ability of individuals to control environmental influences on size will therefore have consequences for their fitness. In eusocial insects, individual males and reproductive females do not have to forage for themselves or control their local environment. Instead, they are reared by nonreproductive siblings (workers) inside colonies. Workers should benefit from controlling the size of sexuals because these sexuals are usually the only means for workers to transmit their genes to future generations. Nevertheless, considerable intraspecific variation exists around mean sexual size in social hymenopterans, even in species with monomorphic sexuals. This variation could result from genetic influences on sexual size, for instance sexuals may be selected to not agree to worker interests, or be due to strong, unpredictable environmental conditions constraining the efforts of workers to control sexual size. In a study that is the first of its kind I investigated genetic and environmental components of sexual body size variation in the ant Lasius niger, examining sexuals from wild colonies with one or several fathers (paternity levels established through microsatellite DNA offspring analysis). Evidence was found for a genetic component of size (broad-sense heritability of up to 42%) but strong common-colony effects (among-colony variation in food availability or in worker capacities to restrain sexual selfishness) also increased the size differences among colonies. Workers thus seem to only have partial-control over sexual size, but may be doing the best of a bad job.

Relationships between phenotype, mating behavior, and fitness of queens in the ant Lasius niger.

Considerable attention has focused on why females of many species mate with several males. For social hymenopteran insects, efforts have primarily concentrated on determining whether multiple mating increases colony performance due to the increased genetic diversity. Most of these studies are correlative because it is difficult or impossible to experimentally mate queens in most species. Thus, the positive associations found between multiple paternity and colony fitness in some cases may not be due to direct effects of genetic diversity but could, in theory, arise from high-quality queens having more mates. Here we show that in the ant Lasius niger variation in the number of matings covaries with queen phenotype. Young queens that were heavier at the time of the mating flight were significantly more likely to mate with several males. As a result, heavier queens stored more sperm. The initial weight of queens was significantly associated with the probability of surviving mating flights during the two years of the study, with queens of intermediate weight having the highest across-year survival. Queen initial weight was also significantly and positively associated with the quantity of brood at the time of the first worker eclosion as well as colony productivity at the time of hibernation. By contrast, there was little evidence for a positive effect of the number of matings on colony performance when the effect of mate number and queen initial weight were considered simultaneously.

Why do some social insect queens mate with several males? Testing the sex-ratio manipulation hypothesis in Lasius niger.

Although multiple mating most likely increases mortality risk for social insect queens and lowers the kin benefits for nonreproductive workers, a significant proportion of hymenopteran queens mate with several males. It has been suggested that queens may mate multiply as a means to manipulate sex ratios to their advantage. Multiple paternity reduces the extreme relatedness value of females for workers, selecting for workers to invest more in males. In populations with female-biased sex ratios, queens heading such male-producing colonies would achieve a higher fitness. We tested this hypothesis in a Swiss and a Swedish population of the ant Lasius niger. There was substantial and consistent variation in queen mating frequency and colony sex allocation within and among populations, but no evidence that workers regulated sex allocation in response to queen mating frequency; the investment in females did not differ among paternity classes. Moreover, population-mean sex ratios were consistently less female biased than expected under worker control and were close to the queen optimum. Queens therefore had no incentive to manipulate sex ratios because their fitness did not depend on the sex ratio of their colony. Thus, we found no evidence that the sex-ratio manipulation theory can explain the evolution and maintenance of multiple mating in L. niger.

Isolation from African Sykes' monkeys (Cercopithecus mitis) of a lentivirus related to human and simian immunodeficiency viruses.

Analysis of serum samples from 100 wild-caught or colony-born Sykes' monkeys (Cercopithecus mitis) in Kenya revealed that 59 animals had antibodies cross-reactive to human immunodeficiency virus type 2 (HIV-2) and to simian immunodeficiency viruses (SIVs). A lentivirus, designated SIVsyk, was isolated from five of six seropositive asymptomatic Sykes' monkeys, but in four cases isolation was possible only after depletion of CD8+ lymphocytes and cocultivation of the CD4(+)-enriched cell population with peripheral blood mononuclear cells from seronegative Sykes' monkeys. SIVsyk resembled other SIVs and HIVs morphologically, had an Mg2(+)-dependent reverse transcriptase enzyme, and replicated in and was cytopathic for CEMx174 and Sup-T1 cells. SIVsyk differred substantially from other SIVs, however, in that it failed to replicate in normal human, mangabey, and macaque peripheral blood mononuclear cells and serum from seropositive Sykes' monkeys immunoprecipitated env antigens from HIV-1 as well as from HIV-2, SIVsmm, and SIVagm. These data demonstrate a high prevalence of natural infection in Sykes' monkeys in Kenya with a lentivirus that appears to be unique with respect to its host range and antigenic cross-reactivity.

Hippocampal damage associated with prolonged and fatal stress in primates.

Sustained exposure to glucocorticoids (GCs), adrenal hormones secreted during stress, can cause neural degeneration in the rat. This is particularly so in the hippocampus, a principal neural target site for GCs, in which GCs can exacerbate the rate of neuron death during normal aging, as well as the severity of neuronal damage after various neurological insults. Thus, stress can be a potent modulator of hippocampal degeneration in the rat. The present report suggests a similar association in the primate. Eight vervet monkeys, housed in a primate center in Kenya, that had died spontaneously from 1984 to 1986, were found at necropsy to have multiple gastric ulcers; a retrospective, neuropathological study was then done of this opportunistic population. Compared with controls euthanized for other research purposes, ulcerated monkeys had marked hippocampal degeneration that was apparent both quantitatively and qualitatively, and both ultrastructurally and on the light-microscopic level. Minimal damage occurred outside the hippocampus. Damage was unlikely to have been due to an agonal or post-mortem artifact. Instead, ulcerated monkeys appear to have been subject to sustained social stress, perhaps in the form of social subordinance in captive breeding groups: most came from social groups, had significantly high incidences of bite wounds at necropsy, and had hyperplastic adrenal cortices, indicative of sustained GC release. Moreover, the specific hippocampal cell fields damaged in ulcerated animals matched those damaged by GCs in the rodent hippocampus. Thus, this represents the first evidence suggesting that sustained stress, via GC hypersecretion, might be neurodegenerative in the primate.

Reproduction in the vervet monkey (Cercopithecus aethiops): III. The menstrual cycle.

The menstrual cycles of 17 multiparous vervet monkeys were studied. Based on estradiol, progesterone, and LH profiles, ovulation is predicted to occur on day 13 of the 32.4-day menstrual cycle. Estradiol peaked on the day preceding the LH peak in 75% of cycles. Average luteal phase length (progesterone greater than 4 nmol/l) was 18 days, with progesterone rising above 4 nmol/l on the day of the LH peak. Vaginal cytology and perianal skin coloration exhibited too much within- and among-animal variability to be reliable indicators of menstrual cycle stages. Uterine biopsies of the proliferative phase were characterized by mild pseudostratification of the columnar epithelium and absence of glandular secretion; in contrast, those of the luteal phase had marked pseudostratification of the tall columnar epithelium with glandular secretions in the lumen. A few follicular-phase samples contained structures such as tortuous uterine glands with secretions. Such structures are more characteristic of the luteal phase. It is suggested that their presence can be explained by incomplete sloughing of the endometrium at menstruation, as this is known to be light or convert in this species.

Serological survey and virus isolation of simian T-cell leukemia/T-lymphotropic virus type I (STLV-I) in non-human primates in their native countries.

Infection with a simian retrovirus (STLV-I) closely related to human T-lymphotropic virus type I (HTLV-I) was investigated in non-human primates living in their native countries in Africa and Asia. Serum antibodies cross-reacting with HTLV-I antigens were detected in 85 of 567 non-human primates of 30 species. Seropositive animals were found among African green monkeys, olive baboons, Sykes' monkeys, mandrills and patas monkeys in several countries in Africa, and cynomolgus monkeys, Celebes macaques and siamangs in Indonesia. The frequency of seropositivity was much higher in adult than in young African green monkeys, cynomolgus monkeys and Celebes macaques. STLV-Is were isolated by establishing II lines of virus-producing lymphoid cells in the presence of interleukin-2 from 5 species of seropositive non-human primates, i.e. the African green monkey, Sykes' monkey, Celebes macaque, cynomolgus monkey and siamang. All these cell lines had T-cell markers and Tac antigen, and the cell lines from the African green monkey and Sykes' monkeys were Leu2a+ while those from other species were Leu3a+. These cell lines expressed viral antigens reacting with human sera from adult T-cell leukemia (ATL) patients and monoclonal antibodies (MAbs) against p19 and p24 of HTLV-I core proteins, and produced virus particles having RNA-dependent DNA polymerase activity. Cellular DNAs from these cell lines contained provirus sequences homologous to HTLV-I, shown by Southern blot hybridization. The restriction patterns of these provirus genomes were different from those of HTLV-I and were also dissimilar in the different species.

Early implantation and embryonic development of the baboon: stages 5, 6 and 7.

Implantation stages of the olive baboon, Papio cynocephalus anubis, showing embryonic development equivalent to Carnegie stages 5, 6 and 7 of development, were collected by hysterotomy and examined histologically. The younger specimens (stage 5) consisted of a thick trophoblastic plate composed of cytotrophoblast and syncytiotrophoblast with multiple small clefts, and a bilaminar disk embryo with a small slit-like amniotic cavity. An epithelial plaque response was present in the uterine epithelium immediately peripheral to the implantation site, within an area of pronounced uterine edema. The bilaminar embryonic disk consisted of columnar epiblast cells underlying the amniotic cavity, and thickened visceral endodermal cells that form part of the yolk sac. The slightly further developed placenta (stage 6) consisted predominantly of cytotrophoblast including primary villi and syncytiotrophoblast lining large spaces containing maternal blood. Secondary placental villi were present in the oldest group (stage 7), and there was modest decidualization of the uterine stroma. An epithelial plaque response persisted, but varied in extent. The sequence of events in early development in the baboon is similar to that in the rhesus monkey insofar as blood space formation and endometrial responses are concerned. However, the plaque response is not so great as in the rhesus; there is no secondary placenta, and the decidual response is slightly more extensive.

Bovine tuberculosis in a wild baboon population: epidemiological aspects.

A follow-up of an outbreak of Mycobacterium bovis in a population of feral baboons in Kenya was undertaken after one year by necropsy of euthanized, clinically ill animals, and tuberculin testing of others. It was concluded that the source of the infection was animals feeding on village slaughterhouse offal of M. bovis infected cows. Secondary (direct) baboon-baboon transfer of the disease appeared to be minimal or nonexistent.

Pregnancy outcome in free-ranging vervet monkeys (Cercopithecus aethiops).

A preliminary assessment of individual female differences in conception rate and fetal wastage has been determined for a population of wild vervet monkeys (Cercopithecus aethiops). One of three troops of vervet monkeys, the subjects of a long-term behavioral study, was trapped and blood was obtained for electrophoretic analysis. Pregnant females exhibited a distinctive serum aminopeptidase phenotype allowing a conclusive determination of pregnancy. Of the seven females diagnosed as pregnant, three later gave birth. Of the females that aborted, two were nulliparous and one was very old. Studies of captive animals have indicated that age and rank may affect a female's ability to carry a fetus to term. These factors, rather than the trapping procedure, may have been responsible for most of the fetal loss in the trapped troop. A comparison of all three troops for a 3-year period indicated that there were fluctuations in yearly birth success of individual females, as well as a relatively high miscarriage rate. The results of this study indicate the advantages of obtaining joint behavioral and biological data.

STLV-I antibodies in feral populations of East African vervet monkeys (Cercopithecus aethiops).

Serum samples from feral populations of African green monkeys (Cercopithecus aethiops) were screened for antibodies to the simian T-lymphotropic virus, type I (STLV-I). Blood samples had been collected from 336 monkeys in 4 regions of central and southern Kenya in 1978 and 1979, from 114 monkeys in central Ethiopia in 1973, and from 85 monkeys from the Kampala region of Uganda in 1966. A total of 178/535 monkeys (33%) were seropositive (STLV-I+). Only 4/114 monkeys (4%) from Ethiopia were seropositive compared to 25/85 Ugandan monkeys (29%) and 149/336 Kenyan monkeys (44%). Epidemiological analysis of the Kenyan monkeys showed that 37% of the males and 54% of the females were STLV-I+, and that there was a progressive increase in the proportion of STLV-I+ monkeys of both sexes with age, rising from an average of 16% in infants (less than 9 months) to an average of 69% in adults (greater than 42 months). The proportion of STLV-I+ monkeys was higher among females in each age category. Seropositivity for antibodies to STLV-I had no apparent effect on the health of monkeys, and no association with the occurrence of Hepatocystis parasitemia was seen in this species. The analysis of data from infants of STLV-I+ mothers showed that seroconversion had occurred in 1 of 3 cases, suggesting that vertical transmission of the STLV-I virus is not an inevitable consequence for infants of seropositive mothers.

Experimental infection of the baboon (Papio cynocephalus) with Echinococcus granulosus of camel, cattle, sheep and goat origin from Kenya.

In different areas of the world, strains of Echinococcus granulosus have been described which appear to vary in their infectivity, and laboratory primates have been used as indicators of their infectivity to man. This phenomenon was evaluated in Kenya for hydatid material of human, camel, cattle, sheep and goat origin. Viable eggs, produced by experimental infections in dogs with larval material from all the above intermediate hosts, were fed separately to four baboons (Papio cynocephalus) in each case. Baboons were autopsied between 373 and 501 days following infection and the liver, lungs, heart, spleen and kidneys were thoroughly inspected. Hydatid cysts were recovered from two baboons in each of the camel, sheep and goat groups, three baboons in the cattle group and none of the baboons in the human group. Fertile cysts were found in the cattle and goat groups and it is suggested that the baboon could be used as an experimental model for this parasite.