Expression of calbindin-D28K by yolk sac and chorioallantoic membranes of the corn snake, Elaphe guttata

The yolk splanchnopleure and chorioallantoic membrane of oviparous reptiles transport calcium from the yolk and eggshell to the developing embryo. Among oviparous amniotes, the mechanism of calcium mobilization to embryos has been studied only in domestic fowl, in which the mechanism of calcium transport of the yolk splanchnopleure differs from the chorioallantoic membrane. Transport of calcium is facilitated by calbindin-D(28K) in endodermal cells of the yolk splanchnopleure of chickens but the chorioallantoic membrane does not express calbindin-D(28K). We used immunoblotting to assay for calbindin-D(28K) expression in yolk splanchnopleure and chorioallantoic membrane of the corn snake, Elaphe guttata, to test the hypothesis that the mechanism of calcium transport by extraembryonic membranes of snakes is similar to birds. High calbindin-D(28K) expression was detected in samples of yolk splanchnopleure and chorioallantoic membrane during late embryonic stages. We conclude that calbindin-D(28K) is expressed in these extraembryonic membranes to facilitate transport of calcium and that the mechanism of calcium transport of the chorioallantoic membrane of the corn snake differs from that of the chicken. Further, we conclude that calbindin-D(28K) expression is developmentally regulated and increases during later embryonic stages in the corn snake.     

Somatic polyploidy in extraembryonic membranes of the snake,Elaphe guttata

Summary Video-densitometric DNA measurements of Feulgenstained tissues of 42 day old eggs of the corn snake,Elaphe g. guttata (Columbridae, Serpentes), revealed a basic DNA content of 2C=2.17 pg, with somatic polyploidy in the allantois, the chorioallontois, the yolk sac, and other extraembryonic membranes. The maximum value determined was 128C (in binucleate cells 2×128C) at the distal pole of the egg. This is the first report of somatic polyploidy in a snake, and one of the first in reptiles in general.     

Effects of temperature and perch diameter on arboreal locomotion in the snake Elaphe guttata

Arboreality is widespread in multiple lineages of snakes and these habitats are important for foraging, escaping predators, and thermoregulation for many species. However, very little is known about factors influencing the arboreal locomotor abilities of snakes. Arboreal performance was assessed in a semi-arboreal snake (Elaphe guttata) using an artificial perch apparatus. Locomotor velocity, body posture, and balance was measured during movement on three perch diameters (3, 6, 10-cm) at three temperatures (10, 20, 30 degrees C). Velocities attained by E. guttata on perches are much slower than those of terrestrial lateral undulation and swimming and somewhat slower than concertina velocities recorded in other species across the same experimental temperatures. At higher temperatures, faster speeds were associated with a more elongated posture. At lower temperatures, snakes displayed a more looped body posture, but still fell more often than at higher temperatures. Our results suggest that temperature has a large influence on both balance and movement by snakes on perches. Although there were no differences in velocities resulting from perch diameter, snakes fell more often from thicker perches. This differs from arboreal velocities attained by limbed vertebrates, which decrease with decreasing perch diameter, suggesting that snakes have a size-relative advantage over limbed animals, such as lizards, when traversing a network of narrow branches. Future studies investigating arboreal locomotion among snakes that vary both phylogenetically and morphologically are needed to assess the potential benefits of limblessness in complex, three-dimensional environments.     

Hepatozoon species (Apicomplexa: Hepatozoidae) of the corn snake,Elaphe guttata (Serpentes: Colubridae) and the pigmy rattlesnake,Sistrurus miliarius barbouri (Serpentes: Viperidae) in south Florida

Hepatozoon guttata n. sp. is described from the corn snake (Elaphe guttata guttata) from south Florida. Gamonts average 14.6 x 4.6 (13-17 x 3.5-6) and are not recurved, with nucleus situated in the second quarter of the gamont. Erythrocyte cytoplasm rarely appears partially dehemoglobinized. The infected cells are usually distorted and are longer than the uninfected erythrocytes but do not differ in width; nuclei are smaller in length and width than those of uninfected cells. Sporogony in Aedes aegypti occurs within the head and the thorax but rarely in the abdomen. Oocysts are spherical to usually ovoid, 87.2 x 75.9 (45-155 x 40-152.5) and contain on average 7.1 (2-32) sporocysts. Sporocysts are spherical to ovoid, 34.8 x 31.0 (20-55 x 17.5-47.5), with 45.7 (14-89) sporozoites contained within. The pigmy rattlesnake (Sistrurus miliarius barbouri) in south Florida is parasitized by H. sistruri n. sp. Gamonts average 14.1 x 5.6 (12.6-15.8 x 4.7-6.3) in breadth and are not recurved, with the nucleus always situated in the second quarter of the gamont commonly at the midbody. Erythrocyte cytoplasm is not dehemoglobinized. The infected cells are always distorted and similar in length to the uninfected cells but with greater width and enlarged nuclei. Sporogony in A. aegypti occurs within head, thorax, and abdomen. Oocysts are spherical to usually ovoid, 163.6 x 154.7 (92-245 x 82-240) and contain 27.4 (12-42) sporocysts. Sporocysts are round to usually ovoid, 39.7 x 33.5 (25-50 x 20-50), with 45.7 (19-70) sporozoites.     

Molecular analysis of the 18S rRNA gene of Cryptosporidium serpentis in a wild-caught corn snake (Elaphe guttata guttata) and a five-species restriction fragment length polymorphism- based assay that can additionally discern C. parvum from C. wrairi

An adult wild-caught corn snake (Elaphe guttata guttata) was presented for humane euthanasia and necropsy because of severe cryptosporidiosis. The animal was lethargic and >5% dehydrated but in good flesh. Gastric lavage was performed prior to euthanasia. Histopathologic findings included gastric mucosal hypertrophy and a hemorrhagic erosive gastritis. Numerous 5- to 7-microm-diameter round extracellular organisms were associated with the mucosal hypertrophy. A PCR, acid-fast stains, Giemsa stains, and an enzyme immunoassay were all positive for Cryptosporidium spp. PCR and restriction fragment length polymorphism (RFLP) analysis on gastric lavage and gastric mucosal specimens, and subsequent sequencing of the 18S rRNA gene, enabled a distinct molecular characterization of the infecting organism as Cryptosporidium serpentis. Until recently, studies on snake Cryptosporidium have relied on host specificity and gross and histopathologic observations to identify the infecting species. A multiple alignment of our sequence against recently published sequences of the 18S rRNA gene of C. serpentis (GenBank accession no. AF093499, AF093500, and AF093501 [L. Xiao et al., unpublished data, 1998]) revealed 100% homology with the C. serpentis (Snake) sequence (AF093499) previously described by Xiao et al. An RFLP method to differentiate the five presently sequenced strains of Cryptosporidium at this locus was developed. This assay, which uses SpeI and SspI, complements a previously reported assay by additionally distinguishing the bovine strain of Cryptosporidium from Cryptosporidium wrairi.     

Phylogeographic analysis of the cornsnake (Elaphe guttata) complex as inferred from maximum likelihood and Bayesian analyses

Most phylogeographic studies have used maximum likelihood or maximum parsimony to infer phylogeny and bootstrap analysis to evaluate support for trees. Recently, Bayesian methods using Marlov chain Monte Carlo to search tree space and simultaneously estimate tree support have become popular due to its fast search speed and ability to create a posterior distribution of parameters of interest. Here, I present a study that utilizes Bayesian methods to infer phylogenetic relationships of the cornsnake (Elaphe guttata) complex using cytochrome b sequences. Examination of the posterior probability distributions confirms the existence of three geographic lineages. Additionally, there is no support for the monophyly of the subspecies of E. guttata. Results suggest the three geographic lineages partially conform to the ranges of previously defined subspecies, although Shimodaira-Hasegawa tests suggest that subspecies-constrained trees produce significantly poorer likelihood estimates than the most likely trees reflecting the evolution of three geographic assemblages. Based on molecular support, these three geographic assemblages are recognized as species using evolutionary species criteria: E. guttata, Elaphe slowinskii, and Elaphe emoryi [phylogeographic, maximum likelihood, maximum parsimony, bootstrap, Bayesian, Markov chain Monte Carlo, cornsnake, Cytochrome b, geographic lineages, E. guttta, E. slowinskii, and E. emoryi].     

Observational learning of tool-use by young chimpanzees

In the current study two groups of young chimpanzees (4–6 and 8–9 years old) were given a T-bar and a food item that could only be reached by using the T-bar. Experimental subjects were given the opportunity to observe an adult using the stick as a tool to obtain the food; control subjects were exposed to the adult but were given no demonstration. Subjects in the older group did not learn to use the tool. Subjects in the younger group who were exposed to the demonstrator learned to use the stick as a tool much more readily than those who were not. None of the subjects demonstrated an ability to imitatively copy the demonstrator's precise behavioral strategies. More than simple stimulus enhancement was involved, however, since both groups manipulated the T-bar, but only experimental subjects used it in its function as a tool. Our findings complement naturalistic observations in suggesting that chimpanzee tool-use is in some sense «culturally transmitted» — though perhaps not in the same sense as social-conventional behaviors for which precise copying of conspecifics is crucial.     

Experimentally reducing clutch size reveals a fixed upper limit to egg size in snakes, evidence from the king ratsnake, Elaphe carinata

Snakes are free of the pelvic girdle's constraint on maximum offspring size, and therefore present an opportunity to investigate the upper limit to offspring size without the limit imposed by the pelvic girdle dimension. We used the king ratsnake (Elaphe carinata) as a model animal to examine whether follicle ablation may result in enlargement of egg size in snakes and, if so, whether there is a fixed upper limit to egg size. Females with small sized yolking follicles were assigned to three manipulated, one sham-manipulated and one control treatments in mid-May, and two, four or six yolking follicles in the manipulated females were then ablated. Females undergoing follicle ablation produced fewer, but larger as well as more elongated, eggs than control females primarily by increasing egg length. This finding suggests that follicle ablation may result in enlargement of egg size in E. carinata. Mean values for egg width remained almost unchanged across the five treatments, suggesting that egg width is more likely to be shaped by the morphological feature of the oviduct. Clutch mass dropped dramatically in four- and six-follicle ablated females. The function describing the relationship between size and number of eggs reveals that egg size increases with decreasing clutch size at an ever-decreasing rate, with the tangent slope of the function for the six-follicle ablation treatment being -0.04. According to the function describing instantaneous variation in tangent slope, the maximum value of tangent slope should converge towards zero. This result provides evidence that there is a fixed upper limit to egg size in E. carinata.     

Spatial learning in golden hamsters: Relationship between food-searching strategies and difficulty of the task

In an elevated maze consisting of three reconvergent radial arms, golden hamsters were tested with the same experimental rule: to choose each path without repeating any choice. However, variations of procedure concerning (a) the location of the reward in the maze, and (b) reinforcement contingencies, were introduced in order to define several problems involving variable levels of difficulty. The relationship between response strategies and difficulty of the task was then studied. The common learning criterion was the achievement of three consecutive correct daily sessions, each session corresponding to a particular sequence (pattern) of choices of paths. Response strategies were studied by analyzing the patterns obtained over the three final sessions in which an animal reached the learning criterion. Such a set of patterns (triplet) could be heterogeneous (patterns all different), mixed (two identical patterns, one different) or stereotyped (identical patterns). No relationship was found between the mean level of difficulty presented by each learning problem and the occurrence of a particular type of triplet. However, in each situation, mixed triplets were the most frequently recorded and corresponded to the medium individual speeds of learning whereas heterogeneous triplets corresponded to rapid successes and stereotyped triplets to delayed successes. These findings indicate that, whatever the problem designed to be tested in a three-arm maze, the various forms of solutions reflect different individual adaptative mechanisms.     

Reinforcing effect for corn oil stimulus was concentration dependent in an operant task in mice

Corn oil is reported to elicit a conditioned place preference (CPP) in a CPP test in mice. To further investigate a reinforcing effect of corn oil, we studied whether the corn oil acts as a reinforcer under a progressive ratio (PR) schedule in the operant task. BALB/c mice were trained to lever press for sucrose and corn oil. After reaching a stable break-point for 100% corn oil, the PR test was conducted for various concentrations of corn oil (0%-100%). The reinforcing effect of corn oil was increased in a concentration-dependent manner under the PR schedule. A mineral oil and 0.3% xanthan gum as vehicles did not show any reinforcing effect in the PR test, suggesting that oily and viscous texture was not related to the reinforcing property of corn oil. The break-point for corn oil was attenuated by pretreatment with (-)-sulpiride, a D(2) antagonist, in the PR test. On the other hand, SCH23390, a D(1) antagonist, did not influence the break-point. Furthermore, the pretreatment with (-)-sulpiride or SCH23390 did not influence the intake of corn oil in a one-bottle test for 30 min, suggesting that the dopaminergic system is involved in the reinforcing effect but not the consumption of corn oil in mice. In conclusion, operant response to corn oil is concentration-dependently enhanced under the PR schedule. This reinforcing effect of corn oil is at least partly mediated through the dopaminergic systems via D(2) receptors.     

Chronic treatment with estrogen receptor agonists restores acquisition of a spatial learning task in young ovariectomized rats

Previous work has shown that continuous estradiol replacement in young ovariectomized rats enhances acquisition of a delayed matching-to-position (DMP) T-maze task over that of ovariectomized controls. The mechanism by which estradiol confers this benefit has not been fully elucidated. This study examined the role of selective estrogen receptor agonists of ERα, ERβ, and GPR30 in the enhancement of spatial learning on a DMP task by comparing continuous estradiol replacement with continuous administration of PPT (an agonist of ERα), DPN (an agonist of ERβ), or G-1 (an agonist of GPR30) relative to gonadally intact and ovariectomized vehicle-treated controls. It was found that ovariectomy impaired acquisition on this task, whereas all ER selective agonists restored the rate of acquisition to that of gonadally intact controls. These data suggest that estradiol can work through any of several estrogen receptors to enhance the rate of acquisition on this task.     

Effects of task complexity on learning to skip steps: an operant analysis

In most response sequences auxiliary responses stop occurring as training increases. Auxiliary responses are precurrent responses that increase the likelihood of reinforcement for subsequent responding, are not required by the programmed contingencies, and occur in situations in which transfer of stimulus control is not prevented. For example, when someone is learning to solve arithmetic problems, some steps, such as writing down intermediate calculations, are skipped as training increases. A paired-associates task was used to investigate the decrease of auxiliary response, in which participants had to learn the second member (arbitrary characters) of each pair upon being presented with the first member (different shapes), and could look up an auxiliary screen (auxiliary response) in order to do so. Task complexity was varied by changing the average programmed frequency of reinforcement for individual responses (Experiment 1) and response sequences (Experiment 3), the programmed probability of reinforcement for responses given a position (PPRPos) with a fixed (Experiment 2) or variable number of associated pairs (Experiment 4), and the programmed probability of reinforcement for responses given a shape with fixed (Experiment 5) or variable (Experiment 6) number of characters per shape. Increases in these variables produced systematic decreases in the duration of auxiliary behavior necessary to learn the task. These results suggest that some aspects of task complexity can be measured based upon the quantification of the programmed contingencies of reinforcement.     

The use of an incremental repeated acquisition task to assess learning in children

The development of valid animal models of learning is especially important since learning is critical for nearly all aspects of human behavior and identifying appropriate surrogates provides additional opportunity to study various aspects of learning. Examining the factors that affect learning is often complicated by the need to administer the same task repeatedly across experimental conditions. Incremental repeated acquisition (IRA) tasks have been used extensively in animal research because they circumvent this problem by requiring a subject to learn different response chains repeatedly across sessions. The present study examined the association of age, sex of the participant, and IQ on the performance of an incremental repeated acquisition task in 837 children, aged 5-13 years. This task required children to learn to press four response levers in a specific sequence that was randomly chosen. Illumination of colored indicator lights signaled position in the required response chain. Initially, for the first link, only one of the four levers was correct: a response to it resulted in the delivery of a monetary reinforcer (5 cents). After mastery of the first link (i.e. three correct presses), the children were presented with a two-link response chain: a different lever had to be pressed before pressing the previously correct lever. After mastery of the two-link chain, the response chain length was once again increased, and so on until a response chain consisting of six links was completed or until the task timed out. Older children and children with higher IQs mastered longer response chain lengths and were more accurate in performance of this learning task than younger children. In addition, older children and children with higher IQs had higher effective response rates and lower ineffective response rates. No significant effects of the sex of the participant were demonstrated for any of the variables on this task, except overall response rate. The results indicate that this test is sensitive to developmental variables in children, with the degree of sensitivity of certain dependent variables being age-dependent. Characterization of performance of this task by humans facilitates comparisons with animal models employing the same task, thus enhancing its translational utility.     

Stimulus enhancement in greylag geese: socially mediated learning of an operant task

We recently observed the spreading of a novel tradition in a flock of semiferal greylag geese, Anser anser: an increasing number of individuals began to bite and chew the stems of butterbur, Petasites hybridus. Because this behaviour spread particularly fast within families, social learning seemed to be involved. We therefore designed an experiment with hand-reared goslings, which were socially imprinted on humans, to investigate whether and how the observation of an experienced tutor affects the acquisition of a novel skill. Goslings had to open the gliding lid of a box to get at a food reward. To each of seven hand-reared observers a human tutor demonstrated where and how to open the lid, whereas seven controls remained untutored. All observers learned to perform the task but only one of the controls succeeded. The observers explored more often at the position shown by the tutor than elsewhere and seemingly learned by trial and error. In contrast, control birds explored primarily at positions that did not allow them to open the box. These results indicate that in greylag goslings the observation of an experienced model facilitates the learning of an operant task. We conclude that stimulus enhancement followed by operant conditioning were the mechanisms involved, which may have accounted for the fast spread of the stem-chewing tradition between family members. Copyright 2000 The Association for the Study of Animal Behaviour.     

Spatial effects, sampling errors, and task specialization in the honey bee

Task allocation patterns should depend on the spatial distribution of work within the nest, variation in task demand, and the movement patterns of workers, however, relatively little research has focused on these topics. This study uses a spatially explicit agent based model to determine whether such factors alone can generate biases in task performance at the individual level in the honey bees, Apis mellifera. Specialization (bias in task performance) is shown to result from strong sampling error due to localized task demand, relatively slow moving workers relative to nest size, and strong spatial variation in task demand. To date, specialization has been primarily interpreted with the response threshold concept, which is focused on intrinsic (typically genotypic) differences between workers. Response threshold variation and sampling error due to spatial effects are not mutually exclusive, however, and this study suggests that both contribute to patterns of task bias at the individual level. While spatial effects are strong enough to explain some documented cases of specialization; they are relatively short term and not explanatory for long term cases of specialization. In general, this study suggests that the spatial layout of tasks and fluctuations in their demand must be explicitly controlled for in studies focused on identifying genotypic specialists.