Further tests of the Scalar Expectancy Theory (SET) and the Learning-to-Time (LeT) model in a temporal bisection task

To contrast two models of timing, Scalar Expectancy Theory (SET) and Learning to Time (LeT), pigeons were exposed to a double temporal bisection procedure. On half of the trials, they learned to choose a red key after a 1s signal and a green key after a 4s signal; on the other half of the trials, they learned to choose a blue key after a 4-s signal and a yellow key after a 16-s signal. This was Phase A of an ABA design. On Phase B, the pigeons were divided into two groups and exposed to a new bisection task in which the signals ranged from 1 to 16s and the choice keys were blue and green. One group was reinforced for choosing blue after 1-s signals and green after 16-s signals and the other group was reinforced for the opposite mapping (green after 1-s signals and blue after 16-s signals). Whereas SET predicted no differences between the groups, LeT predicted that the former group would learn the new discrimination faster than the latter group. The results were consistent with LeT. Finally, the pigeons returned to Phase A. Only LeT made specific predictions regarding the reacquisition of the four temporal discriminations. These predictions were only partly consistent with the results.     

Manipulating pre-feed, density of reinforcement, and extinction produces disruption in the Location variation of a temporal discrimination task in pigeons

When pharmacological and non-pharmacological agents are used to disrupt temporal discrimination, two major findings have emerged in the literature. One result reveals lateral shifts of the psychophysical curve for time due to disruptors, while the other is a decrease in accuracy for classifying short and long intervals and a flattening of the psychophysical curve. These results represent a discrepancy within the timing literature that requires clarification. The current study determined the effects of pre-feed, increased density of reinforcement during session, and extinction on the Location variation of a temporal discrimination procedure. The results showed that extinction and pre-feed (at higher levels), when presented in an acute fashion, led to right-ward shifts in the psychophysical curve. Our results, when compared to similar studies in the literature, suggest that lateral shifts are more likely to be found due to disruptors when the Location variation is being used and when procedures are less complicated.     

Feature-short and feature-long discrimination learning in the pigeon: conditional control of a two-event temporal map

Eight pigeons were trained to peck an illuminated target key on discrete-trial fixed-interval schedules of reinforcement by food. Four birds were exposed to a feature-short (FS) task where a feature light signaled shortening of the forthcoming target-outcome interval from 30 to 15s, while the other four birds were exposed to a feature-long (FL) task where a feature light signaled extension of the forthcoming target-outcome interval from 15 to 30s. The discrimination performance measured by differential temporal distributions of pecks between featured and non-featured target trials suggested that the target-food temporal map was under conditional control of the feature light in both groups. The FS discrimination was more difficult to learn than the FL discrimination. This FS inferiority implies that our birds did not resort on the simple temporal discrimination by timing from the trial onset. The simple temporal discrimination account was also negated by the finding that increasing the feature-target gap did not have a predicted effect on the response distribution.     

Interstimulus interval as a discriminative stimulus: Evidence of the generality of a novel asymmetry in temporal discrimination learning

Three appetitive conditioning experiments with rats examined temporal discrimination learning within Pavlovian conditioning trials. In all experiments, the duration of a feature white noise stimulus signaled whether or not a subsequent 10-s target tone would be reinforced. In Experiment 1, the feature durations were 4 and 1 min. For one group of rats (Group 4+/1−), 4 min of noise signaled that the tone would be reinforced and 1 min of noise signaled that the tone would not be reinforced. A second group (Group 1+/4−) was trained with the reverse contingency. The results showed a clear asymmetry in temporal discrimination learning: rats trained with 4+/1− (Long+/Short−) learned the discrimination readily (responding more in the tone on reinforced than on nonreinforced trials), whereas rats trained with 1+/4− (Short+/Long) did not. In Experiment 2, the feature durations were shortened to 60 and 15 s. Due to strong excitatory conditioning of the 15-s feature, the reverse asymmetry was observed, with the Short+/Long− discrimination learned more readily than the Long+/Short− discrimination. However, Experiment 3 demonstrated that the original Long+/Short− advantage could be recovered while using 60− and 15-s feature durations if the excitatory conditioning of the feature was reduced by including nonreinforced feature trials. The results support previous research involving the timing of intertrial intervals and are consistent with the temporal elements hypothesis which holds that the passage of time is encoded as a series of hypothetical stimulus elements.     

The role of temporal generalization in a temporal discrimination task

Two experiments trained rats to discriminate two or three stimulus durations using a temporal discrimination task. A standard peak shift effect was observed when training was administered with short versus long signals in Experiment 1. Both discrimination accuracy scores and the generalization gradients revealed that shorter intervals were discriminated more accurately, which may be due to the scalar property of timing. In Experiment 2, three signals (short, medium, and long) were associated with three different responses, or two of the intervals were associated with one response (short and long or short and medium) and the other interval with a different response. Here, the short/medium versus long discrimination was learned most readily of the three tasks. The results of both experiments indicated a strong contribution of learning of individual durations combined with scalar generalization gradients, but Experiment 2 indicated that categorical encoding of durations may have also been operating.     

Song recognition by temporal cues in a group of closely related bushcricket species (genus Tettigonia)

Female phonotaxis of Tettigonia viridissima and T. caudata was investigated on a walking compensator to determine the temporal parameters of the male song used for song recognition, and to compare them with the previously described pulse rate filtering of T. cantans. The T. cantans song is continuous with a ≈30-Hz pulse rate. The T. caudata song has a higher pulse rate (≈40 Hz) and duty cycle than T. cantans and a distinct verse structure. The T. viridissima song is continuous with a double-pulse pattern. While the pulse rate is essential for song recognition in T. cantans, neither pulse rate not verse structure were essential for song recognition in T. caudata: females responded to signals above a minimum duty cycle. T. viridissima females did not require the double-pulse structure, but a single long pulse, equivalent to the duration of the double pulses and interval between them, was effective. Song attractiveness was limited by a minimum duration of the merged double pulse, and by minimum and maximum duration of the interval between them. Pulse rate recognition had little if any importance in either of the species investigated. Thus, the three congeners use different mechanisms for temporal song recognition. Accepted: 18 June 1998     

雄性高原鼠兔长鸣时间分配及其影响因素

长鸣是高原鼠兔常见的鸣声类型之一。了解高原鼠兔长鸣发生的时间分配规律及其影响因素对于理解其生存策略具有重要意义。本研究从2018年2月至2019年2月,采用录音机记录其鸣叫声音与单反相机录制鸣叫行为相结合的方法,在青藏高原东缘的甘肃省玛曲县记录了高原鼠兔一年内长鸣发生的时间规律,并研究其长鸣发生频率与外界非生物因素(温度和风速)以及内在生理因素(睾酮和孕酮)的关系。结果表明:(1)高原鼠兔长鸣由雄性个体发出,主要出现在3—5月的繁殖季节,每只个体每日长鸣总时长为130～180 s,每日长鸣主要发生在日出后2 h和日落前2 h内;(2)高原鼠兔繁殖期长鸣时长与睾酮水平呈正相关。当睾酮水平为40 pg/mL左右时,长鸣时长达到1000 s,睾酮水平下降到36 pg/mL以下时,长鸣时长只有150 s左右。非繁殖期长鸣发生与气温和风速呈负相关。综述,高原鼠兔雄性个体全年长鸣发生呈现明显的季节性,繁殖季长鸣与睾酮水平增高密切相关。     

Spatial and temporal differences in the community structure of endophytic fungi in the carnivorous plant Pinguicula moranensis (Lentibulariaceae)

According to our hypothesis, temporal heterogeneity due to seasonality (summer rosette vs. winter rosette) instead of spatial heterogeneity, showed higher influence on both the community and the infection rate of endophytes present in the carnivorous plant Pinguicula moranensis. In addition, the infection rate of endophytic fungi isolated from roots was higher than that observed in leaves although the endophytic community was similar between both tissues. Overall, we isolated endophytic fungi members of Dothideomycetes, Leotimocyetes, Eurotiomycetes and Sordariomycetes. P. moranensis harbors at least 10 genera, seven of them (Astrocystis sp., Bipolaris sp., Cercospora sp., Coniochaeta sp., Ijuhya sp., Neomollisia sp. and Ochroconis sp.) reported for the first time in carnivorous plants. We advanced some implications about the role that these endophytes could be playing in this carnivorous plant, illustrating a certain level of specificity in the endophytic community of fungi present in leaves and roots.     

A characterization model with spatial and temporal resolution for life cycle impact assessment of photochemical precursors in the United States

Background, aim, and scope  Traditional life cycle impact assessment methodologies have used aggregated characterization factors, neglecting spatial and temporal variations in regional impacts like photochemical oxidant formation. This increases the uncertainty of the LCA results and diminishes the ease of decision-making. This study compares four common impact assessment methods, CML2001, Eco-indicator 99, TRACI, and EDIP2003, on their underlying models, spatial and temporal resolution, and the level at which photochemical oxidant impacts are calculated. A new characterization model is proposed that incorporates spatial and temporal differentiation. Materials and methods  A photochemical air quality modeling system (CAMx-MM5-SMOKE) is used to simulate the process of formation, transformation, transport, and removal of photochemical pollutants. Monthly characterization factors for individual US states are calculated at three levels along the cause–effect chain, namely, fate level, human and ecosystem exposure level, and human effect level. Results and discussion  The results indicate that a spatial variability of one order of magnitude and a temporal variability of two orders of magnitude exist in both the fate level and human exposure and effect level characterization factors for NO_x. The summer time characterization factors for NO_x are higher than the winter time factors. However, for anthropogenic VOC, the summer time factors are lower than the winter time in almost half of the states. This is due to the higher emission rates of biogenic VOCs in the summer. The ecosystem exposure factors for NO_x and VOC do not follow a regular pattern and show a spatial variation of about three orders of magnitude. They do not show strong correlation with the human exposure factors. Sensitivity analysis has shown that the effect of meteorology and emission inputs is limited to a factor of three, which is several times smaller than the variation seen in the factors. Conclusions  Uncertainties are introduced in the characterization of photochemical precursors due to a failure to consider the spatial and temporal variations. Seasonal variations in photochemical activity influence the characterization factors more than the location of emissions. The human and ecosystem exposures occur through different mechanisms, and impacts calculated at the fate level based only on ozone concentration are not a good indicator for ecosystem impacts. Recommendations and perspectives  Spatial and temporal differentiation account for fate and transport of the pollutant, and the exposure of and effect on the sensitive human population or ecosystem. Adequate resolution for seasonal and regional processes, like photochemical oxidant formation, is important to reduce the uncertainty in impact assessment and improve decision-making power. An emphasis on incorporating some form of spatial and temporal information within standard LCI databases and using adequately resolved characterization factors will greatly increase the fidelity of a standard LCA. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Determination of ovulation and pregnancy in the marmoset (Callithrix jacchus) by monitoring of urinary hydroxypregnanolone excretion

In Callithrix jacchus the urinary excretion of hydroxypregnanolone (5 alpha-pregnane-3 alpha, 7 alpha-diol-20-one) shows a luteal rise during the ovulatory cycle. This progesterone metabolite can therefore be used as an indication of ovulation, implantation, and subsequently for the persistence of the pregnancy. The excretion can be monitored by high performance thin-layer chromatography (HPTLC) and postchromatographic derivatization after enzymatic hydrolysis of the conjugate. Profiles of urinary hydroxypregnanolone levels are presented and correlated with luteinizing hormone excretion. The method is noninvasive and therefore suited for long-term studies in these monkeys.     

Spatial and temporal patterns in the recruitment of the intertidal barnacle Chthamalus malayensis Pilsbry (Crustacea: Cirripedia) on the equatorial shores of Peninsular Malaysia and Singapore

Spatial and temporal variation in the recruitment of the intertidal barnacle Chthamalus malayensis was examined over one year (September 2003-August 2004) on the equatorial shores of southern Peninsular Malaysia and Singapore. A nested sampling design was applied for the first time on these shores, over three spatial scales - hundreds of kilometres, kilometres to tens of kilometres and tens of metres - and temporal variation was determined through monthly sampling of recruits. Shores within 2° north of the equator on the East and West coasts of southern Peninsular Malaysia and the southern coast of Singapore were selected as study sites. Generally, all three coasts recruited throughout the year, with varying intensities. There was a clear regional pattern, where the largest number of recruits was found on the East Coast of Malaysia, and the least in Singapore. Analyses of variance (ANOVA) showed that variability occurred on the smallest scale, but only in nine of the twelve months examined, consequently resulting in significant temporal and spatial interaction. Calculated variance components indicated that small-scale variation accounted for most of the overall variability. The potential causes of the spatial and temporal patterns of C. malayensis recruitment, and implications on tropical-temperate comparisons will be discussed.     

Olfactory discrimination of eight food flavors in the capuchin monkey (Cebus apella): Comparison between fruity and fishy odors

Operant conditioning was used to investigate how primates discriminate between odor qualities. Eight artificial food flavors, selected from either a “Fishy” or “Aroma/Fruity” category, were used. During the presence of one of the two odors S+ or S−, the monkey was reinforced by pushing a response key when S + was presented. The tufted capuchins discriminated most accurately when both odors were Aroma. Discrimination was more accurate when S+ was Fruity odor and S− was Fishy odor. When both odors were Fishy, discrimination could not be acquired in 20 sessions. All of the flavors used, except apple, were novel for the subjects, which suggests that capuchins can innately discriminate among them. The data also suggest that Aroma odors are more salient than Fishy odors. The results also suggested an innate aversion to Fishy odors.     

Cholesterol as a factor regulating the influence of natural (PAF and lysoPAF) vs synthetic (ED) ether lipids on model lipid membranes

In this work we have performed a comparative study on the effect of antineoplastic ether lipid-edelfosine (ED), its natural analogs — Platelet Activating Factor (PAF) and its precursor (lyso-PAF), both lacking anticancer properties, on cholesterol/phosphatidylcholine (Chol/PC) monolayers, serving as model membranes. Since all the above ether lipids are membrane active, it can be expected that their effect on membranes may differentiate their biological activity. Our investigations were aimed at studying potential relationship of the effect of ED, PAF and lyso-PAF on model membranes, differing in condensation. We have modified molecular packing of Chol/PC model systems either by increasing the level of sterol in the system or changing the structure of PC, while keeping the same sterol content. Additionally, we have performed a detailed comparison of the miscibility of ED, PAF and lyso-PAF with various membrane lipids. The collected data evidenced that all the investigated ether lipids influence Chol/PC films in the same way; however, in a different magnitude. Moreover, the interactions of ED, PAF and lyso-PAF with model membranes were the strongest at the highest level of sterol in the system. A thorough analysis of the obtained results has proved that the effect of the investigated ether lipids on membranes is not dependent on the condensation of the system, but it is strongly determined by the concentration of cholesterol. Since ED was found to interact with model membranes stronger than PAF and lyso-PAF, we have suggested that this fact may contribute to differences in cytotoxicity of these compounds.     

Mathematical modelling of morphogenesis in fungi: a key role for curvature compensation ('autotropism') in the local curvature distribution model

The assumption that the mushroom stem has the ability to undergo autonomic straightening enables a mathematical model to be written that accurately mimics the gravitropic reaction of the stems of Coprinus cinereus . The straightening mechanism is called curvature compensation here, but is equivalent to the 'autotropism' that often accompanies the gravitropic reactions of axial organs in plants. In the consequently revised local curvature distribution model, local bending rate is determined by the difference between the 'bending signal' (generated by gravitropic signal perception systems) and the 'straightening signal' (proportional to the local curvature at the given point). The model describes gravitropic stem bending in the standard assay with great accuracy but has the virtue of operating well outside the experimental data set used in its derivation. It is shown, for example, that the mathematical model can be fitted to the gravitropic reactions of stems treated with metabolic inhibitors by a change of parameters that parallel the independently derived physiological interpretation of inhibitor action. The revised local curvature distribution model promises to be a predictive tool in the further analysis of gravitropism in mushrooms.     

Diel and seasonal variation of a molluscan taxocoenosis associated with a Zostera marina bed in southern Spain (Alboran Sea)

The diel and seasonal variation of molluscs living in a Zostera marina bed (12–14 m depth) from southern Spain have been studied for one year using a small Agassiz trawl for collecting the samples (222 m²). The frequent and dominant species were very similar in both diurnal and nocturnal samples, including mainly gastropods such as Jujubinus striatus, Nassarius pygmaeus, Mitrella minor, Calliostoma planatum, Rissoa membranacea or Smaragdia viridis. Nevertheless, a significant increase of abundance of scavengers (e.g. Nassarius spp.) and carnivores (e.g. cephalopods) was registered in nocturnal samples. The abundance was maximal in spring and summer in diurnal and nocturnal samples and also in autumn for nocturnal ones, displaying significantly higher values in nocturnal samples. The species richness, diversity and evenness displayed a similar seasonal trend for diurnal and nocturnal samples, with maximum values during summer months. Monthly variation of the molluscan composition (species presence-absence data) was more acute than diel variation, according to the Cluster, MDS and ANOSIM results. Nevertheless, both monthly and diel changes in the structure (species abundance data) of the molluscan taxocoenosis were important throughout the year. Diel changes in the structure of the molluscan fauna are related to an increase of abundance of some species at nighttime due to vertical movements from the sediment to the shoots or along them (e.g. J. striatus, Nassarius spp.) or due to horizontal movements from adjacent habitats (e.g. cephalopods). Nevertheless, some species such as Rissoa spp. or Bittium spp. stay on the leaves of Z. marina during day as well as nighttime. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Spatial and temporal distribution of the zoobenthos community in a woodland pond (Switzerland)

Spatial and temporal changes of zoobenthos composition and density were assessed in a woodland pond, near Geneva (Switzerland), by monthly sampling (during 15 months), on the three main substrates: a submerged macrophyte (Chara sp.), an emergent macrophyte (Typha latifolia stems) and allochthonous detritus (oak leaves).Many taxa showed preferences for one or two of the substrates, as expressed by differences in densities, but few taxa presented an exclusivity for one substrate. Zoobenthos densities (per m² of pond bottom) were largely dominated on all substrates and seasons by Chironomidae and Oligochaeta. Other dense macroinvertebrates included Ostracoda (on Chara and Typha stems), Gastropoda (on Typha stems), Ephemeroptera (all substrates), and Ceratopogonidae (on Typha stems).The magnitude of temporal differences in densities was expressed by the ratio of maximal to minimal monthly densities and reached values between 2.8 to 11.8. On all substrates the highest monthly densities were observed in summer or beginning of autumn. The temporal changes in densities were strongly influenced by the life cycles of the invertebrates: presence of numerous young individuals in summer (for example Cypridopsis vidua, Cloeon dipterum, Caenis horaria, Ferrissia wautieri), and emergence (for example Chironomidae in April). Fluctuations in densities, especially on Chara and leaves, were also attributed to modification of substrate condition (surface availability, stage in the decomposition or senescence process). Temporal fluctuations of densities were compared with other lentic water bodies: highest densities showed a common trend, occurring between end of spring and beginning of autumn.     

The role of pigeon consumption in the population dynamics and breeding performance of a peregrine falcon (<Emphasis Type="Italic">Falco peregrinus</Emphasis>) population: conservation implications

In this paper, we describe and analyze the diet of peregrine falcons during a long-term period (1982–2002). A combination of direct observations of prey brought to nests, prey remains, and regurgitated pellets were used to calculate diet diversity and dietary overlap between peregrine pairs. We also examined diet diversity in relation to breeding performance. All peregrine pairs fed mainly on birds, with pigeons the most common prey. An increase in pigeon availability has been associated with both an increase in population size and an increase in breeding performance (measured as the average productivity of pairs per year) of a small peregrine falcon population in eastern Spain. Average productivity was lower when dietary breadth was higher. We speculate that our results were the synergistic effect of declining persecution and increased pigeon availability through increased popularity of keeping racing pigeons. There is a conflict of interests between pigeon fanciers and peregrine conservation. As a consequence, this could result to an increased risk of mortality by direct persecution. In accordance with this, conservation measures aimed at preventing direct persecution are encouraged.     

Fitness consequences of pheromone production and host selection strategies in a tree-killing bark beetle (Coleoptera: Curculionidae: Scolytinae)

Timing of arrival at a resource often determines an individual’s reproductive success. Tree-killing bark beetles can reproduce in healthy trees by attacking in adequate numbers to overcome host defences that could otherwise be lethal. This process is mediated by aggregation and antiaggregation pheromones. Beetles that arrive early in such a “mass attack” must contend with undiminished tree defences, and produce enough pheromones to attract more beetles, but have a head start on gallery construction and egg-laying. Beetles that arrive late may be impeded by competition and diminishing availability of phloem, but should experience fewer costs associated with pheromone production and battling tree defences. We investigated relationships between timing of arrival, body size, pheromone production and fitness in the southern pine beetle, Dendroctonus frontalis. In field experiments, we captured beetles that arrived early (pioneers) and late on slash pine trees, Pinus elliottii, and measured pheromone amounts in their hindguts. We marked gallery entrances of beetles as they landed on a tree and measured their reproductive success after the attack terminated. We found no difference in body size or pheromone amounts between early and late arrivers. Most beetles arrived at the middle of the attack sequence, and excavated longer galleries per day than early arrivers. The number of offspring produced per day by beetles that established galleries midway through mass attack was higher than those that arrived early or very late in the sequence. Our results suggest that beetles do not exhibit adaptive phenotypic plasticity in pre-landing pheromone production, depending on the extent of previous colonisation of a host. Rather, it appears that stabilising selection favours beetles that attack in the middle of the sequence, and contributes to attack synchrony. Synchronous attack on trees is essential before population booms characteristic of tree-killing bark beetles can occur in nature.     

Comparison of the predicted in vivo behaviour of the Sn(II)-APDDMP complex and the results as studied in a rodent model

In a quest for more effective radiopharmaceuticals for pain palliation of metastatic bone cancer, this paper relates results obtained with ((117m)Sn labelled) Sn(II) complexed to the bone seeking bisphosphonate, N,N-dimethylenephosphonate-1-hydroxy-3-aminopropylidenediphosphonate (APDDMP). APDDMP is synthesised from the known bone cancer pain palliation agent 1-hydroxy-3-aminopropylidenediphosphonate (APD, Pamindronate). This work is performed to utilise the idea that the low bone marrow radio toxicity of (117m)Sn could afford a highly effective radiopharmaceutical in pain palliation but also in the curative treatment of bone metastasis. Complex-formation constants of APDDMP with the important blood plasma metal-ions, Ca(2+), Mg(2+), Zn(2+) as well as the added metal ion, Sn(2+) were measured by glass electrode potentiometry at 25 degrees C and I = 150 mM. Blood plasma models were constructed using the computer code ECCLES and the results compared with those gathered from tests on a rodent model. The ((117m)Sn-labelled) Sn(II)-APDDMP complex was found to have only some liver and bone uptake although a high trabecular to normal bone ratio was recorded. From the blood plasma model this was shown to be primarily due to the high affinity of APDDMP for Ca(II) causing some of the Sn(II)-APDDMP complex to dissociate. High kidney uptake and excretion as well as high bladder uptake was recorded which was shown to be due to the dissociation of the Sn(II)-APDDMP complex in blood plasma. Animal model observations could be explained by the blood plasma modelling.     

Brood defense and brood size in the great tit (Parus major): a test of a model of unshared parental investment

To understand the interaction of the many contextual variablesthat affect parental behavior a number of static optimalitymodels have been developed. Among these the one by Lazarus andInglis (1986) is the only one to specifically predict the magnitudeof unshared parental investment (PI), i.e., of parental carethat carries a cost to the parent and that benefits all currentoffspring equally because it cannot be divided among them. Weinvestigated specifically how parent great tits (Parus major)gear their brood defense, a form of unshared PI, to the sizeof the brood at stake and to the risk incurred as a functionof the type of predator. The predators used were dummies ofthe great spotted woodpecker (Picoides major) and of the tawnyowl (Strix aluco). Normally, adults can approach the woodpeckerwith impunity; it had inflicted heavy losses to nestlings ofthe study populations of great tits near Wolfsburg, Lower Saxony.Parent great tits whose brood had been artificially reducedto two young responded to the dummy with less defense than dida control group with their pre-test brood size left intact.The nature of defense was qualitatively the same as that elicitedby a live woodpecker. Parents confronting the owl near theirbrood decreased their response with an artificial reductionin brood size much less. Because the owl used poses a seriousrisk to the defenders, as compared to the woodpecker, the resultlends powerful support to the "total loss" version of the modelof unshared PI; it predicts brood size to affect unshared PImore strongly when there is less risk to the parent. This interpretationis correct to the extent that one premise of the model, namelythat of uncompromised parentage, can be relaxed; great tit broodscontain a sizeable number of extrapair young. Males defendedtheir brood more strongly than did females. Sex and brood manipulationadded up linearly when affecting defense level, i.e., therewas no interaction.