The behaviour of Bulinus africanus: A circadian profile

Six different behaviour patterns have been studied across the circadian spectrum in the tropical freshwater snail Bulinus africanus. Snails entrained to a circadian light/dark regimen show a diurnal rhythm of locomotor activity, and feeding, photokinetic responsiveness, egg laying and hatching all follow a similar pattern. The rhythms are endogenous and persist under constant conditions. In contrast defaecation occurs mainly during the dark period. This antiphase relationship may be due in part to a cycle of absorption and fragmentation in the digestive gland cells, but it is likely that other factors are involved.     

Defaecation by African elephants (Loxodonta africana africana (Blumenbach))

A study of defaecation in the African elephant was carried out at the Voi headquarters of the Tsavo (East) National Park. Four orphaned animals aged between 1 and 10 years were observed for 4 days and 3 nights. During the day the time and weight of each individual's defaecation was recorded while at night only the time of defaecation. Details of all the records are presented. Analysis has shown that the amount of dung produced with each defaecation bears a similar characteristic to that of the growth curve of these animals. The rate of defaecation does not vary significantly with age. Records of defaecation arranged by time demonstrate an apparent periodicity with a low peak mid-morning and a high peak mid-afternoon. The potential use of this information in feeding and population studies is discussed.     

Feeding and defaecation behaviour of Triatoma patagonica (Del Ponte, 1929) (Hemiptera: Reduviidae)

Among the vectors of Chagas disease, Triatoma patagonica is a species in the process of adaptation to the human environment being recently registered in urban and suburban zones. However, its importance as a vector of Chagas disease is unknown. The aim of this work was to evaluate two aspects of vectorial competence: the feeding behaviour and the defaecation pattern. These processes were studied in females of T. patagonica fed ad libitum on a restrained pigeon. The results showed that the blood meal size was negatively correlated with the time of first defaecation (r = -0.42). The first defaecation was emitted before the first 10 min and defaecations during feeding were frequent. A total of 73% of females, defaecated during the first 30 min post-feeding. These results suggest that if this species subsequently colonizes the domicile, it would be capable of transmitting Trypanosoma cruzi.     

The twenty-four hour activity cycle of captive coypus (Myocastor coypus)

The twenty-four hour activity cycle of captive coypus was investigated by direct visual observation, automatic recording and by measuring the amount of faeces eliminated at hourly intervals. Coypus were active from just before sunset and the "Zeitgeber" or cue for activity onset may be declining light. The activity period usually ended before sunrise and the interval between the end of the period and dawn was inversely related to temperature. The animals fed and defaecated throughout the active period and 80–86 % of the faecal pellets were produced in water. Refection started shortly after the end of the feeding period, when the animals had returned to the nest, and continued until an hour or two after midday.
A model is suggested which involves two passages of food through the gut and a cycle length of 24 hours from ingestion to defaecation. The feeding and defaecation periods were equal in duration, but the refection period was 40% shorter. Hypothetically this may be due to a caecum by-pass mechanism during the second passage of food through the gut so that the defaecation period, like the feeding period, is controlled by the rate at which food passes through the pyloric sphincter. In contrast the period of refection may be controlled by the rate at which food remains are voided from the caecum. Coypus spent the pre-dawn and daylight hours on nests and in contact with familiar conspecifics. These behaviours may have the function of temperature maintenance in the coldest part of the night and of avoiding diurnal predators.     

The feeding ecology of Axiothella rubrocincta (Johnson) (Polychaeta: Maldanidae)

The feeding ecology of Axiothella rubrocincta (Johnson) from Tomales Bay, California, is described. This worm inhabits a U-shaped tube of agglutinated sand grains and mucus. Morphological adaptations such as nuchal and anal plaques prevent the tube from becoming clogged. Foreign debris entering the tube is either consumed or incorporated into the tube wall. The proboscis and notosetae are probably used to clean the tube wall. A. rubrocincta combines feeding and burrowing activities to form the funnel and complete the tube: it does not ingest sediment while burrowing. Organic matter deposited into the funnel is buried there by sand slides originating at the rim of the funnel. The concentration of organic matter within the funnel is significantly higher than for non-funnel sediments. A. rubrocincta consumes food from the upper 2 cm of the substratum and is 4.6% efficient as a depositfeeder. It also feeds within the funnel and can ingest large quantities of food. This feeding process is described. A. rubrocincta irrigates its tube at a rate of 5.1 ml sea water/g/h while feeding, and briefly reverses this current to a rate of 0.1 ml/g/h when defaecating. The rhythmic activity patterns are integrated: the mean defaecation and inverse pumping time is 14.4 min at 15 ± 1 C.     

Behavioural coordination between an aphis (Symydobius oblongus von Heyden; Hemiptera: Callaphidae) and the ant that attends it (Formica lugubris Zetterstedt; Hymenoptera: Formicidae): An ethological analysis

The aphis Symydobius oblongus von Heyden can coordinate its defaecation with the presence of its attendant Formica lugubris Zetterstedt, so that 86% of droplets are produced in the 14% of time when the aphis is attended. The behaviour of aphids and ants was recorded in the field and subjected to sequence analysis and temporal analysis. A method of temporal analysis using the 1st-nth order pre-and post-event histogram (PPEH) is described. The results of these analyses support the view that coordination of defaecation and attendance is the result of two processes of communication. The commonest is the response of the aphis to the arrival of an attendant, probably mediated by contact of the ant's antennae and palps with the aphis. Less frequently, aphids ‘call’ ants, perhaps signalling their readiness to defaecate by movements of the abdomen which the ants detect visually.     

Feeding by the planktonic calanoid copepod Acartia clausi Giesbrecht on natural suspended particulate matter of varying quantity and quality

The defaecation rate of females of the calanoid copepod Acartia clausi Giesbrecht as a measure of the ingestion rate was measured in relation to quantity and quality of natural suspended particulate matter. In the April–September experiments, phytoplankton was the dominant component of suspended particulate matter and the ambient chlorophyll a concentration was high, ranging from l.11 to 23.91 μg·l⁻¹. The defaecation rate became saturated below the ambient particle concentration and the weight-specific defaecation rate was rectilinearly fitted against particle concentration. In terms of chlorophyll a, the critical concentration for saturation of the weight-specific defaecation rate ranged from 0.616 to 1.710 μg·l⁻¹. In the October–November experiments, the ambient chlorophyll a concentration ranged from 0.27 to 0.53 μg·l⁻¹ and non-living particles became the dominant component. No critical concentration was observed and the slope of the line describing the relationship between particle concentration and weight-specific defaecation rate was greatly reduced in comparison with that obtained in the April–September experiments. The reduction of the slope may be due to the dominance of non-living particles.     

Defaecation in the stem nematode, Ditylenchus dipsaci

The process of defaecation in the stem nematode, Ditylenchus dipsaci , has been analysed using cinemicrography. During feeding regular shortenings of the hind end of the body are accompanied by groups of a few defaecation periods. Each period begins as the shortened hind body re-elongates, and consists of several defaecation cycles. In each cycle the posterior rectum becomes dorsally bowed and, as it straightens, the rectal valve, rectum and anus open. Faeces pass out irregularly while the rectum remains open and are forcibly expelled as it closes. Defaecation in D. dipsaci is intermediate between that of the high-pressure nematode Aphelenchoidzs blastophthorus and the low-pressure passive feeder Hexatylus viviparus. We suggest that the main function of defaecation in plant nematodes is to excrete excess water.     

A quantitative description of the behaviour of wild juvenile plaice (Pleuronectes platessa L.)

The behaviour of young plaice (Pleuronectes platessa L.) as they migrate up and down sandy beaches with the tide is described. Their behaviour during this migration consists mainly of swimming and feeding interspersed with rarer behavioural acts. Over short periods swimming behaviour can be described by a random model in which the probability of a swim occurring remains constant. This probability varies markedly, however, from hour to hour. Two types of swimming movement are recognized: one of very short duration represents searching for food and the other, longer, category serves to transport the fish up and down the shore. Variations in the feeding rate and in the frequency, duration, and direction of swimming movements over the tidal cycle are described and related to the changing physical and biological conditions that the fish experience during their intertidal movements.     

Variables Controlling Entry into and Exit from the Steady-State,One of Two Modes of Feeding in Aplysia

Background

Aplysia feeding is a model system for examining the neural mechanisms by which changes in motivational state control behavior. When food is intermittently present, Aplysia eat large meals controlled by a balance between food stimuli exciting feeding and gut stimuli inhibiting feeding. However, when food is continuously present animals are in a state in which feeding is relatively inhibited and animals eat little. We examined which stimuli provided by food and feeding initiate steady-state inhibition of feeding, and which stimuli maintain the inhibition.

Results

Multiple stimuli were found to control entry into the steady-state inhibition, and its maintenance. The major variable governing entry into the steady-state is fill of the gut with bulk provided by food, but this stimulus cannot alone cause entry into the steady-state. Food odor and nutritional stimuli such as increased hemolymph glucose and L-arginine concentrations also contribute to inhibition of feeding leading to entry into the steady-state. Although food odor can alone cause some inhibition of feeding, it does not amplify the effect of gut fill. By contrast, neither increased hemolymph glucose nor L-arginine alone inhibits feeding in hungry animals, but both amplify the inhibitory effects of food odor, and increased glucose also amplifies the effect of gut fill. The major variable maintaining the steady-state is the continued presence of food odor, which can alone maintain the steady-state for 48–72 hrs. Neither increased glucose nor L-arginine can alone preserve the steady-state, although they partially preserve it. Glucose and arginine partially extend the effect of food odor after 72 hrs.

Conclusions

These findings show that control of Aplysia feeding is more complex than was previously thought, in that multiple inhibitory factors interact in its control.     

Molecular analysis of photic inhibition of blood-feeding in Anopheles gambiae

Background

Anopheles gambiae mosquitoes exhibit an endophilic, nocturnal blood feeding behavior. Despite the importance of light as a regulator of malaria transmission, our knowledge on the molecular interactions between environmental cues, the circadian oscillators and the host seeking and feeding systems of the Anopheles mosquitoes is limited.

Results

In the present study, we show that the blood feeding behavior of mosquitoes is under circadian control and can be modulated by light pulses, both in a clock dependent and in an independent manner. Short light pulses (~2–5 min) in the dark phase can inhibit the blood-feeding propensity of mosquitoes momentarily in a clock independent manner, while longer durations of light stimulation (~1–2 h) can induce a phase advance in blood-feeding propensity in a clock dependent manner. The temporary feeding inhibition after short light pulses may reflect a masking effect of light, an unknown mechanism which is known to superimpose on the true circadian regulation. Nonetheless, the shorter light pulses resulted in the differential regulation of a variety of genes including those implicated in the circadian control, suggesting that light induced masking effects also involve clock components. Light pulses (both short and long) also regulated genes implicated in feeding as well as different physiological processes like metabolism, transport, immunity and protease digestions. RNAi-mediated gene silencing assays of the light pulse regulated circadian factors timeless, cryptochrome and three takeout homologues significantly up-regulated the mosquito's blood-feeding propensity. In contrast, gene silencing of light pulse regulated olfactory factors down-regulated the mosquito's propensity to feed on blood.

Conclusion

Our study show that the mosquito's feeding behavior is under circadian control. Long and short light pulses can induce inhibition of blood-feeding through circadian and unknown mechanisms, respectively, that involve the chemosensory system.     

A novel flexible method to split feeding behaviour into bouts

Before meal patterns can be analysed properly, a biologically relevant meal criterion must be determined in order to group short-term feeding behaviour into meals. Existing methodologies are based on modelling of the frequency distributions of intervals between feeding events but these methods cannot be used if the proper distributions cannot be clearly identified. For such cases we developed two new methods – (1) based on the analysis of the distribution of between-meal interval lengths only and (2) based on the analysis of changes in the probability of animals starting to feed with time since the last feeding event. Both methods were developed using a data set of over 700,000 records of visits to feeders obtained with broilers (Gallus gallus) aged between 2 and 5 weeks. The two methods resulted in meal criteria estimates of 20.1 and 17.5 min, respectively, which, when applied to the data set, gave statistically significant but very small differences in meal characteristics. The new methods were tested against an independent cow (Bos taurus) data set and the resultant meal criteria compared with those predicted by an existing method. The two novel methods estimated meal criteria for cows at 27.9 and 35.5 min, compared with 28.9 min for the existing method. Again, these differences in meal criteria resulted in only very small differences in meal characteristics. Even though meal criteria were relatively similar for birds and cows, characteristics such as average daily number of meals (10.9 and 5.9), meal size (12.5 g and 7.4 kg) and meal duration (7.7 and 31.4 min) were very different. The analyses show that, if the appropriate distributions of intervals cannot be identified, meal criteria can still be estimated for species as diverse as mature cows and young birds by the novel methodologies developed here.     

The temporal structure of feeding behavior

Meals have long been considered relevant units of feeding behavior. Large data sets of feeding behavior of cattle, pigs, chickens, ducks, turkeys, dolphins, and rats were analyzed with the aims of 1) describing the temporal structure of feeding behavior and 2) developing appropriate methods for estimating meal criteria. Longer (between-meal) intervals were never distributed as the negative exponential assumed by traditional methods, such as log-survivorship analysis, but as a skewed Gaussian, which can be (almost) normalized by log-transformation of interval lengths. Log-transformation can also normalize frequency distributions of within-meal intervals. Meal criteria, i.e., the longest interval considered to occur within meals, can be estimated after fitting models consisting of Gaussian functions alone or of one Weibull and one or more Gaussian functions to the distribution of log-transformed interval lengths. Nonuniform data sets may require disaggregation before this can be achieved. Observations from all species were in conflict with assumptions of random behavior that underlie traditional methods for criteria estimation. Instead, the observed structure of feeding behavior is consistent with 1) a decrease in satiety associated with an increase in the probability of animals starting a meal with time since the last meal and 2) an increase in satiation associated with an increase in the probability of animals ending a meal with the amount of food already consumed. The novel methodology proposed here will avoid biased conclusions from analyses of feeding behavior associated with previous methods and, as demonstrated, can be applied across a range of species to address questions relevant to the control of food intake.     

Successful Predation of Filamentous Bacteria by a Nanoflagellate Challenges Current Models of Flagellate Bacterivory

Current models suggest that (i) filamentous bacteria are protected against predation by nanoflagellates, (ii) prey size is positively correlated with prey-predator contact probability, and (iii) contact probability is mainly responsible for size-selective predation by interception-feeding flagellates. We used five strains of filamentous bacteria and one bacterivorous nanoflagellate, Ochromonas sp. strain DS, to test these assumptions. The five strains, including one spirochete and four Betaproteobacteria strains, were isolated by the filtration-acclimatization method. All five strains possess flexible cells, but they differ in average cell length, which ranged from 4.5 to 13.7 μm. High-resolution video microscopy was used to measure contact, capture, and ingestion rates, as well as selectivity of the flagellate feeding. Growth and feeding experiments with satiating and nonsatiating food conditions, as well as experiments including alternative well-edible prey, were performed. In contrast to predictions by current models, the flagellate successfully consumed all the tested filamentous strains. The ingestion rate was negatively correlated with bacterial length. On the other hand, the lengths of the filamentous bacteria were not positively correlated to the contact rate and capture rate but were negatively correlated to ingestion efficiency. In experiments including alternative nonfilamentous prey, the flagellates showed negative selection for filamentous bacteria, which was independent of food concentration and is interpreted as a passive selection. Our observations indicate that (i) size alone is not sufficient to define a refuge for filamentous bacteria from nanoflagellate predation and (ii) for the investigated filamentous bacteria, prey-predator contact probability could be more influenced by factors other than the prey size.     

Modelling the effect of feeding-related mortality on the feeding strategy of tsetse (Diptera: Glossinidae)

Abstract. Free-living haematophagous insects risk death through host grooming responses or through increased susceptibility to predation whenever they take a bloodmeal. In this paper we investigate the effects of these risks on the feeding strategy of tsetse. A model is presented that allows for death of tsetse by starvation if they do not succeed in feeding within a fixed time (set at 6 days in the first instance) and for mortality specifically associated with feeding. In addition there is background mortality that applies to all flies at all times.
The model is used to compute the individual life-time fertility (number of female puparia per female) as a function of the probability of obtaining a meal (indicated by field data to be very high, usually > 0.85 per day) and the day on which flies start to search for a meal. We suggest that the feeding strategy that would be selected for is that which allows the maximum reproductive output. The model shows that this strategy involves making no attempts to feed for 3–4 days after the previous meal and then attempting to feed with the greatest possible probability until a meal is obtained. The predicted feeding interval, obtained independently of any trapping data, agrees closely with all previous estimates from field studies using a variety of methods. Preliminary results from a laboratory experiment reveal an increased risk of predation of recently fed as compared with hungry tsetse. The lower the actual feeding mortality the more frequently will flies be able to feed should conditions so demand. It is adaptive, however, for tsetse to delay attempting to feed for as long as they can, which is made possible by the near certainty of locating and feeding on a host within 1 day, using their sophisticated sensory systems.     

Antibodies and coinfection drive variation in nematode burdens in wild mice

Coinfections with parasitic helminths and microparasites are highly common in nature and can lead to complex within-host interactions between parasite species which can cause negative health outcomes for humans, and domestic and wild animals. Many of these negative health effects worsen with increasing parasite burdens. However, even though many studies have identified several key factors that determine worm burdens across various host systems, less is known about how the immune response interacts with these factors and what the consequences are for the outcome of within-host parasite interactions. We investigated two interacting gastrointestinal parasites of wild wood mice, Heligmosomoides polygyrus (nematode) and Eimeria spp. (coccidia), in order to investigate how host demographic factors, coinfection and the host’s immune response affected parasite burdens and infection probability, and to determine what factors predict parasite-specific and total antibody levels. We found that antibody levels were the only factors that significantly influenced variation in both H. polygyrus burden and infection probability, and Eimeria spp. infection probability. Total faecal IgA was negatively associated with H. polygyrus burden and Eimeria spp. infection, whereas H. polygyrus-specific IgG1 was positively associated with H. polygyrus infection. We further found that the presence of Eimeria spp. had a negative effect on both faecal IgA and H. polygyrus-specific IgG1. Our results show that even in the context of natural demographic and immunological variation amongst individuals, we were able to decipher a role for the host humoral immune response in shaping the within-host interaction between H. polygyrus and Eimeria spp.     

Using Latent Selection Difference to Model Persistence in a Declining Population

Population persistence is a direct measure of the viability of a population. Monitoring the distribution of declining populations or subpopulations over time can yield estimates of persistence, which we show can be modeled as a latent selection difference (LSD) contrasting attributes of sites where populations have persisted versus those that have not. Predicted persistence can be modeled with predictor covariates to identify factors correlated with species persistence. We demonstrate how to model persistence based on changes in occupancy that can include adjustments for detection probability. Using a known historical distribution of the western grebe (Aechmophorus occidentalis), we adapted methods originally developed for occupancy modeling to evaluate how environmental covariates including emergent vegetation and human developments have affected western grebe persistence in Alberta. The relative probability of persistence was correlated with the extent of shoreline bulrush (Scirpus lacustris), which is important vegetation for nesting cover. We also documented that western grebe populations were less likely to persist on lakes in the boreal forest, primarily located on the northern boundary of the species'' range. Factors influencing occupancy were different than those determining persistence by western grebes; persistence and occupancy were not correlated. Persistence was more likely on lakes with recreational development, reflecting reliance by grebes on the larger, fish-bearing waterbodies that also are attractive for lakeshore development. Unfortunately, the correlation with recreational development on Alberta''s lakes puts grebes at risk for loss of brood-rearing habitats—primary threats to altricial birds—if steps are not taken to prevent disturbance to bulrush stands. Identifying factors related to the persistence of a species—especially one in decline—is a fundamental step in conservation management.     

The use of laser capture microdissection to study the infection of Glycine max (soybean) by Heterodera glycines (soybean cyst nematode)

The soybean cyst nematode (Heterodera glycines) is an obligate parasite of soybean (Glycine max). It is the most destructive pathogen of G. max, accounting for approximately 0.46–0.82 billion dollars in crop losses, annually, in the U.S. Part of the infection process involves H. glycines establishing feeding sites (syncytia) that it derives its nourishment from throughout its lifecycle. Microscopic methods (i.e., laser capture microdissection [LCM]) that faithfully dissect out those feeding sites are important improvements to the study of this significant plant pathogen. Our isolation of developing feeding sites during an incompatible or a compatible reaction is providing new ways by which this important plant-pathogen interaction can be studied. We have used these methods to create cDNA libraries, clone genes and perform microarray analyses. Importantly, it is providing insight not only into how the root is responding at the organ level to H. glycines, but also how the syncytium is responding during its maturation into a functional feeding site.Key words: soybean, Glycine max, soybean cyst nematode, SCN, Heterodera glycines, microarray, gene expression, plant pathogen, parasite, laser capture microdissection     

Experimental Infection of Ornithodoros erraticus sensu stricto with Two Portuguese African Swine Fever Virus Strains. Study of Factors Involved in the Dynamics of Infection in Ticks

African swine fever (ASF) is a frequently devastating hemorrhagic disease of domestic pigs and wild boar and Ornithodoros erraticus sensu stricto argasid ticks are the only biological vectors of African swine fever virus (ASFV) known to occur in Europe. Recently this disease emerged in Eastern Europe and Russian Federation, showing a huge potential for a rapid spread between countries. There is some risk of re-emergence of ASF in the countries where these ticks exist, that can contribute for the persistence of infection and compromise control measures. In this study we aimed to identify factors that determine the probability of infection and its dynamics in the tick vector Ornithodoros erraticus sensu stricto, with two Portuguese strains of ASFV. Our results suggest that these ticks have a high likelihood of excreting the two haemadsorbing ASF viruses of different host origins and that, in field surveys, the analysis of adults and 5^th nymphal stage can provide the best chance of detecting virus infection. The results also indicate that infection of pigs with highly virulent ASF viruses will promote higher rates of infection and a higher likelihood for virus excretion by ticks. Nevertheless, there is also a risk, although lower, that ticks can become infected on pigs that have overcome the acute phase of infection, which was simulated in our study by membrane feeding ticks with low titres of virus. We believe these results can be valuable in designing and interpreting the results of ASF control programmes, and future work can also be undertaken as our dataset is released under open access, to perform studies in risk assessment for ASFV persistence in a region where O. erraticus sensu stricto ticks are present.     

Specifying the Probability Characteristics of Funnel Plot Control Limits: An Investigation of Three Approaches

Background

Emphasis is increasingly being placed on the monitoring and comparison of clinical outcomes between healthcare providers. Funnel plots have become a standard graphical methodology to identify outliers and comprise plotting an outcome summary statistic from each provider against a specified ‘target’ together with upper and lower control limits. With discrete probability distributions it is not possible to specify the exact probability that an observation from an ‘in-control’ provider will fall outside the control limits. However, general probability characteristics can be set and specified using interpolation methods. Guidelines recommend that providers falling outside such control limits should be investigated, potentially with significant consequences, so it is important that the properties of the limits are understood.

Methods

Control limits for funnel plots for the Standardised Mortality Ratio (SMR) based on the Poisson distribution were calculated using three proposed interpolation methods and the probability calculated of an ‘in-control’ provider falling outside of the limits. Examples using published data were shown to demonstrate the potential differences in the identification of outliers.

Results

The first interpolation method ensured that the probability of an observation of an ‘in control’ provider falling outside either limit was always less than a specified nominal probability (p). The second method resulted in such an observation falling outside either limit with a probability that could be either greater or less than p, depending on the expected number of events. The third method led to a probability that was always greater than, or equal to, p.

Conclusion

The use of different interpolation methods can lead to differences in the identification of outliers. This is particularly important when the expected number of events is small. We recommend that users of these methods be aware of the differences, and specify which interpolation method is to be used prior to any analysis.