Effects of nectar concentration on butterfly feeding: measured feeding rates for Thymelicus lineola (Lepidoptera: Hesperiidae) and a general feeding model for adult Lepidoptera

Summary Field observations of the adult European skipper, Thymelicus lineola (Ochs), feeding on concentrated nectars (40–65% sucrose) from a variety of flower species led us to question recent literature stating that butterflies feed primarily, and most effectively, on dilute nectars. Rate of sucrose solution intake, volume consumed and feeding duration were measured for males and females at 25 and 35°C under laboratory conditions. As sucrose concentration increased, the volume of solution ingested per meal first increased and then decreased gradually, while sucrose intake was highest at concentrations 40%. Females fed more than males at all concentrations >10% while temperature had no significant effect on meal size. Feeding duration increased with concentration, was shorter at 35 than at 25°C, and was longer for females than males.The rate of volume intake decreased as concentration incresed, but not nearly as rapidly as predicted by earlier models. Rates did not differ between the sexes but were faster at 35 than 25°C. This increase was contributed to equally by a reduction in viscosity and an increase in power output of the cibarial pump. The form of the relations was similar, with maximum rate of sucrose intake occurring at 40% sucrose.A new mathematical model was developed to describe the rate — concentration relation based on the Hagen-Poiseuille equation for laminar fluid flow through pipes. Our model differs from previous models principally in that the power output of the insect's cibarial pump remains relatively constant while the pressure drop created by the pump to induce suction is highly variable. This change results in a very different feeding rate — sucrose concentration function with the optimal rate of sucrose intake at a concentration of approximately 40%. The model indicates that the same relation should hold for a wide range of proboscis shape and size and type of suction pump, and should therefore be applicable to all other nectar feeders with sucking mouth parts. Independent verifications of the model were carried out by measuring the rate of uptake of sucrose solutions of the adult common armyworm, Pseudaletia unipuncta (Haw.), and of human subjects using a volumetric pipette, both of which gave an excellent fit.Nectar concentrations which correspond to optimal rates of sucrose intake should be highly preferred by insects with high feeding costs, those which are time-limited, or which are very vulnerable while feeding. High transport costs and severe water stress may shift preferences to higher and lower concentrations respectively.     

Sugar concentration and timing of feeding affect feeding characteristics and survival of a parasitic wasp

The availability of food sources is important for parasitoid survival, especially for those that inhabit ecosystems where nectar and honeydew are spatially or temporally scarce. Therefore, the value of even a single meal can be crucial for survival. Psyttalia lounsburyi is a parasitoid, and biological control agent, of the olive fruit fly, Bactrocera oleae. In order to improve our understanding of the basic nutritional ecology of P. lounsburyi and its role in survival we evaluated the effect of a single sucrose meal on the longevity of female and male wasps. We measured the duration of feeding, volume ingested, sucrose consumption, energy content, and longevity of wasps provided with different concentrations of sucrose (0.5, 1, and 2 M) at different times after emergence (0, 1, 2 or 3 days after emergence). Our results showed that longevity was significantly influenced by sucrose concentration and timing of feeding. For females, feeding on sucrose increased the likelihood of survival to varying degrees, ranging from 32.3% to 95.4%, compared to water-only controls. The longest duration of feeding was observed for the highest sucrose concentrations and oldest wasps. The amount of sugar ingested and energy uptake increased, up to a point, as sugar concentration increased. Our results suggest that P. lounsburyi derived greatest benefit from the intermediate concentration (1 M) of sucrose provided 2 or 3 days after emergence. Our study emphasizes the importance of finding balance between increasing longevity and limiting the duration of feeding, and concomitant uptake of nutrients, that is fundamental for survival of the wasp in nature.     

Intravital microscopy and image analysis of Rhodnius prolixus (Hemiptera: Reduviidae) hematophagy: The challenge of blood intake from mouse skin

Hematophagous insects transmit many of the most dangerous parasitic diseases. The transmission usually occurs during hematophagy or just after as this is when the vector and the host are in contact. The contact time is determined by the feeding performance of the insect in each host. In triatomines, feeding performance interferes with both their life cycle and the vectorial competence to transmit the hemoflagellate Trypanosoma cruzi. Triatomine bugs are vessel feeders, obtaining their blood meals directly from the vessels (venules or arterioles) of their vertebrate hosts. The host blood intake rate is not constant during the feeding, and the sucking frequency of triatomines tends to be higher and to contain fewer interruptions in pigeons than in mice. To identify the difficulties encountered by triatomine bugs in obtaining blood meals from mouse skin, we used intravital microscopy techniques associated with electromyograms of the cibarial pump. To monitor the vibration of the cannulated vessels and the blood flow through the head of the insect during the engorgement phase, we introduced a novel method for image analysis. The mean number of vessels used during a Rhodnius prolixus blood meal was 3.4 ± 1.2, and the insects fed more in venules (63%) than in arterioles (37%). An important increase in vascular permeability was observed throughout the feeding. Platelet aggregation, rolling and leukocyte adherence were analyzed on the venular endothelium, showing remarkable increases for some time following the R. prolixus feeding. The reduction in sucking frequency that was observed during insect feeding was likely due to the increased cibarial pump filling time. The monitoring of the vessel wall pulsation also permitted the registration of regurgitation-like movements during blood pumping, with these movements being recorded mostly during the second half of the feeding. The evaluation of blood flow through the head of the insect suggested that the regurgitation-like movements were not true regurgitations and were caused by abrupt difficulties in the function of the cibarial pump. The role of the platelet plugs and the changes in blood viscosity at the R. prolixus feeding site are discussed. The method introduced in the present study to analyze the images brings new insights into the interaction between hematophagous vectors and their hosts, reinforcing the importance of insect saliva throughout the feeding process.     

A relationship between weight loss during food deprivation and subsequent meal size in the locust, Chortoicetes terminifera

Sizes of meals of 0·5 M sucrose taken by adults of either sex of Chortoicetes terminifera showed a strong positive correlation with the weight loss of the insects during the preceding 24 hr period of food deprivation. The relationship was less marked in males fed water, and there was no relationship in those fed 1·0 M sucrose. Meal size in females fed water was as strongly weight loss dependent as in those fed 0·5 M sucrose, but was less so in those fed 1·0 M sucrose. The results indicated that changes occurring after the completion of gut emptying influence meal size.     

Sensory adaptation and the regulation of meal size in the Australian plague locust, Chortoicetes terminifera

Adults of Chortoicetes terminifera consumed larger meals of sucrose solution, applied as drops directly to their mouthparts, than of water. Meal size increased with increasing sucrose concentration up to 0·5 M but meals of 1·0 M sucrose were of lower volume. Locusts fed so that there was a short interval between each successive drop of sugar solution consumed greater volumes than those fed continuously, which indicates that sensory adaptation plays a part in bringing about the termination of meals of sugar solutions. Meals of water were not increased in size by discontinuous application.     

Mechanical determinants of nectar-feeding energetics in butterflies: muscle mechanics,feeding geometry,and functional equivalence

Summary We develop a mechanistic model for nectar feeding in butterflies that integrates the two basic components of the feeding process: the fluid dynamics of nectar flow through the food canal and the contractile mechanics of the muscular, cibarial pump. We use the model to predict the relation between rate of energy intake during feeding and nectar concentration. We then identify nectar concentations that maximize energy intake rates (the optimal concentrations). We illustrate the model using measurements of the food canal and cibarium of Pieris butterflies. The model predicts an overall optimal range of nectar concentration of 31–39% sucrose for butterflies, which is in agreement with previously reported laboratory values. The model also predicts an interaction among the geometries of the food canal, the cibarial cavity, and the cibarial muscles, that allows us to identify the combinations of food canal, cibarium, and muscle dimensions that yield the highest rates of energy intake. Nectar-feeding is functionally equivalent in butterflies and hummingbirds: two physically different feeding mechanisms can yield identical energy intake rates. This equivalence results from a mathematical and physical similarity between quasi-steady-state fluid flow in hummingbrid tongues and the force-velocity characteristics of contracting cibarial muscle in butterflies.     

Feeding behaviour of morphologically similar Rhodnius species: influence of mechanical characteristics and salivary function

Despite their morphological similarities, very similar Rhodnius species (R. prolixus, R. robustus, R. nasutus and R. neglectus) displayed a distinct feeding behaviour when fed on artificial feeder, pigeon or mouse. On pigeon hosts, these species showed distinct groups in terms of cumulative probing time - quicker species (R. prolixus and R. neglectus) followed by R. nasutus and finally a much slower species (R. robustus). On mouse hosts, R. nasutus showed quicker probing time compared to the other three species. Moreover, R. prolixus displayed quicker probing time compared to R. robustus and R. neglectus. Except for R. nasutus, the mean total ingestion rate tended to have different values between feeding sources (artificial feeder>pigeon>mouse). The volume ingested by each cibarial pump contraction and maximum frequency obtained using the artificial feeder are expected to be related to intrinsic mechanical characteristics of the insect feeding apparatus. However, probing time and the modulation of cibarial pump frequency on live hosts may be related to salivary function. R. prolixus showed high mechanical and salivary efficiency, achieving high values of total ingestion rate when fed on artificial feeder or either of the hosts. Comparative analysis suggests that species which possess higher total ingestion rates tend to achieve higher nutritional status, allowing them to reach higher densities.     

An energy budget for juvenile thick-lipped mullet, Crenimugil labrosus (Risso)

A series of experiments were carried out to construct an energy budget for juvenile thick lipped mullet, Crenimugil labrosus Risso. A partial factorial experimental design was used to examine the effects of temperature, fish size and meal size on growth. The maximum ration that the fish were able to ingest completely per day was found to be 0·8, 1·4 and 2·3% wet body weight (b.w.) at 13,18 and 23°C, respectively. Ingested maintenance requirements (M.R.) were estimated to be 137, 205 and 288 cal fish^-1 day^-1 at 13, 18 and 23°C, respectively. At 18deg; C, M.R. varied as 25 W^1.04 cal day^-1, where W= fish weight (g). Growth rate increased with increasing temperature. Maximal conversion efficiency was 21–24% and was achieved closer to the maximum ingested ration with increasing temperature. The relationship between respiration rate and W at 18deg; C for 3-20 g fish is described by: respiration rate (ml O₂ h^-1) = 0·128 W^0.976 The energy cost of apparent specific dynamic action at 18deg; C was found to vary between 5·1% and 23·6% of the calorific value of the ingested meal (1% wet b.w.) , mean (± S.E.)=10·2 ± 2·0%. Post mortem analyses of groups of fish fed 0·2, 0·8 and 1·5% wet b.w. meals showed a significant increase in total lipid and a significant decrease in water content with increasing ratio size. A negative correlation was found between body water content and total lipid (and calories). The mean assimilation efficiency (±s.e.) for 5–10 g mullet at 18deg; C was 73·9 ± 3·6%. The observations reported in this study were brought together to construct an energy budget for juvenile C. labrosus which was found to give a reliable prediction (within 10%) of energy demand and growth under the prevailing experimental conditions. Both gross (K₁) and net (K₂) growth efficiencies, based on energy values, increased with increasing ratio size up to satiation and were independent of temperature. The maximum values of K₁ and K₂ observed were 0·33 and 0·46, respectively. The third order efficiency (K₃) appeared to be independent of temperature and ration size; mean values ranged between 0·66 and 0·84.     

An excitatory state generated during feeding in the locust, Chortoicetes terminifera

Various sequences of 2 μl drops of 1·0 M sucrose and water were applied directly to the mouthparts of restrained adults of the locust Chortoicetes terminifera which had been deprived of all food for 24 hr, and the number of drops consumed was recorded. Locusts fed either single drops of sucrose solution and water in alternation or single drops of sucrose solution alternated with groups of four consecutive drops of water, consumed larger total volumes than locusts fed only drops of water or locusts offered water after having been fed partially or wholly to repletion with 1·0 M sucrose. Towards the end of meals consisting of repeated cycles of one drop of sucrose solution and four drops of water, locusts consumed the one or two drops of water which followed a sucrose drop more quickly than the drop which preceded it. Furthermore, a few locusts which received this treatment terminated their meal while either a drop of sucrose solution or the water drop which immediately followed the sucrose drop was in contact with their mouthparts. The findings provide strong evidence that stimulation received during ingestion of sucrose induces, in addition to an immediate excitatory effect on the feeding system, a perseverating state of neural excitation which, for a short time afterward, increases the chance that the insect will ingest water.     

Effects of sucrose on blood avidity in mosquitoes.

Within 10 min after engorging on 10% sucrose, most females of Aedes aegypti do not seek a human blood meal, but remain quiescent and unresponsive to a human hand in a 1 ft³ cage. The duration of this inhibition in blood avidity varies greatly among individuals, but may last for 2 to 5 hr after drinking sucrose. There was no specific correlation between abdominal distension and blood avidity. When females engorged on varying concentrations of sucrose, it was found that as the concentration increased, fewer mosquitoes would take blood when it was offered after 1 hr. Only 25% of the females which engorged on 0·5 to 1 M sucrose took human blood at this time. As the concentration of sucrose increases, there is a marked decrease in spontaneous flight activity during the first hour after feeding. When unfed females are injected with 10% sucrose or trehalose, none of them took blood for 3 hr, whereas 50% of the saline-injected controls fed on blood.     

Sucking pump activity in feeding behaviour regulation in carpenter ants

Modulation of liquid feeding-rate would allow insects to ingest more food in the same time when this was required. Ants can vary nectar intake rate by increasing sucking pump frequency according to colony requirements. We analysed electrical signals generated by sucking pump activity of ants during drinking solutions of different sucrose concentrations and under different carbohydrate-deprivation levels. Our aim was to define parameters that characterize the recordings and analyse their relationship with feeding behaviour.Signals showed that the initial and final frequencies of sucking pump activity, as well as the difference between them were higher in sugar-deprived ants. However, these parameters were not influenced by sucrose solution concentration, which affected the number of pump contractions and the volume per contraction. Unexpectedly, we found two different responses in feeding behaviour of starved and non-starved ants depending on concentration. Starved ants drank dilute solutions for the same length of time as non-starved ants but ingested higher volumes. While drinking the concentrated solutions, starved ants drank the same volume, but did so in a shorter time than the non-starved ones. Despite these differences, for each analysed concentration the total number of pump contractions remained constant independently of sugar-deprivation level. These results are discussed in the frame of feeding regulation and decision making in ant foraging behaviour.     

Nectar feeding by the hovering hawk moth Macroglossum stellatarum: intake rate as a function of viscosity and concentration of sucrose solutions

Although nectar feeding in insects has long been studied, the knowledge of the effect of nectar energy content on the ingestion dynamics separately from the viscosity of the fluid is very limited. To determine the effects of both factors on the feeding behavior of the hovering hawk moth Macroglossum stellatarum, we developed a method to independently manipulate sucrose concentrations and viscosity. The intake rate was analyzed as a function of sucrose concentration, the concentration at constant viscosity (kept constant by adding tylose, an inert polysaccharide), and of the different viscosities of a 30% weight/weight (w/w) sucrose solution (by adding different amounts of tylose). By increasing the concentration, and thus its viscosity, the solution intake rate (in microl s (-1)) decreased beyond a 20% w/w sucrose solution. For a 30% sucrose solution, the intake rate decreased with increasing viscosity. At constant viscosity, the solution intake rate decreased beyond a 30% w/w sucrose solution. However, if we considered the quantity of sucrose ingested per unit time (sucrose intake rate), the same fitted maximum was attained for both series in which the sucrose concentration changed (33.6% w/w). Results suggest that the gustatory input affects the dynamics of fluid ingestion separately from the viscosity.     

The feeding process of Cimex lectularius (Linnaeus 1758) and Cimex hemipterus (Fabricius 1803) on different bloodmeal sources

The bedbugs Cimex lectularius and Cimex hemipterus are obligate hematophages in all their nymphal instars as well as in the adult stage. The efficiency with which the insects obtain blood from their hosts is directly related to their population dynamics. In the present study we compared the feeding process and salivary content in individuals of these two species when fed on different blood sources or host sites, using a cibarial pump electromyogram. Females ingested more blood than males but needed longer contact time with the host to complete the meal. The bedbug C. lectularius was more efficient than C. hemipterus in obtaining blood from mice and pigeons. With regard to the feeding site on mice, it was easier for the insects to obtain blood from the skin of the belly than that of the back. Individuals of C. hemipterus were able to maintain the cibarial pump functioning at higher frequencies for longer periods when fed on pigeons treated with anticoagulant. Although saliva from C. lectularius contained more hemeproteins and showed more anti-clotting activity its total protein content was similar to that of C. hemipterus. Overall, C. lectularius obtains a bloodmeal more efficiently from its hosts, which may have enabled this species to reach higher levels of infestation than C. hemipterus.     

Why are there no small species among xylem-sucking insects?

Xylem-sucking herbivores must cope with negative tension and extremely low nutrient concentration of xylem fluid. We test the hypothesis that the energetic cost of xylem fluid extraction grows with decreasing body size, as small species have relatively high metabolic rates and small feeding apparatus. The suction pressure gradient needed for feeding (P), calculated using Poiseuilles Law, and the cibarial pump load (L), estimated as xylem fluid volume per unit volume of cibarial muscles, were used to quantify energetic costs associated with feeding. Interspecific relationships between body size and frontoclypeus (cibarial pump) volume, proboscis length and food canal diameter were quantified using the structural relations model and model I (ordinary) regressions applied on species as data points (phylogeny ignored), and on independent (intrageneric) contrasts (taxonomy used as a surrogate of phylogeny). All the dimensions of feeding apparatus changed approximately isometrically with body volume. In consequence, the suction pressure gradient P (body volume)–0.21, and the cibarial pump load L (body volume)–0.17. These relationships result in an allometric change in energetic costs associated with xylem feeding. Energy needed to overcome the resistance of feeding apparatus (P) becomes negligible for any species longer than about 8mm, and there is an important decrease in the L values in species over about 17mm in body length. These allometric trends can result in large species having a wider range of potential (energetically profitable) host plants than small species. Analysis of body size distributions in xylem-feeding insects, which all belong to the Auchenorrhyncha (Hemiptera), showed that xylem feeders tend to be larger than phloem feeders, especially in terms of the minimum body size. Two evolutionary transitions from phloem to xylem feeding in Hemiptera were associated with an increase in the minimum body size. A possibility that the evolution of minimum body size in xylem feeders is energetically constrained is discussed.     

Nectar feeding by the ant Camponotus mus: intake rate and crop filling as a function of sucrose concentration

In independent assays, workers of the ant Camponotus mus were conditioned to visit an arena where they found a large drop of sucrose solution of different concentrations, from 5 to 70% weight on weight (w/w). Single ants were allowed to collect the sucrose solution ad libitum, and feeding time, feeding interruptions, crop load, and intake rates were recorded. Feeding time increased exponentially with sucrose concentration, and this relationship was quantitatively described by the increase in viscosity with concentration corresponding to pure sucrose solutions. Ants collecting dilute solutions (5 to 15% w/w) returned to the nest with partial crop loads. Crop filling increased with increasing sucrose concentration, and reached a maximum at 42.6% w/w. Workers collecting highly concentrated solutions (70% w/w) also returned to the nest with a partially-filled crop, as observed for dilute solutions. Nectar intake rate was observed to increase with increasing sucrose concentration in the range 5 to 30% sucrose. It reached a maximum at 30.8%, and declined with increasing sucrose concentration. Results suggest that both sucrose concentration and viscosity of the ingested solution modulate feeding mechanics as well as the worker's decision about the load size to be collected before leaving the source.     

Physiology of feeding-preference patterns of female black blowflies (Phormia regina Meigen): Modification in responsiveness to salts subsequent to salt feeding

Behavioural experimentation on Phormia regina shows that females previously fed only sucrose, and thus in a state of yeast preference, will ingest single-salt-sucrose mixtures in volumes similar to that of 10% yeast (Rachman et al., 1982). In particular, females will ingest NaCl 0.1 M in 0.2 M sucrose in volumes not statistically different from 10% yeast. In the experiments reported here, groups of females were given ad libitum access to both 0.2 M sucrose and NaCl 0.1 M in 0.2 M sucrose from days 0–4 after eclosion. Potassium and sodium salts were added to 0.2 M sucrose over a spectrum of concentrations and presented to the flies on day 6. The flies rejected all single-salt-sucrose mixtures offered. Even when the pre-test sodium salt was changed to a potassium salt during days 0–4, the flies still rejected the single-salt-sucrose mixtures offered subsequently. However, when sodium and potassium salts were mixed together in 0.2 M sucrose, females ingested the solutions in amounts similar to 10% yeast. Several different concentrations of the double-salt-sucrose solutions were ingested in volumes not statistically different from 10% yeast. Results support the underlying hypothesis that activity in the salt-sensitive chemosensory neurones is involved in protein preference.     

Filtration and ingestion rates of Salpa fusiformis Cuvier (Tunicata : Thaliacea): Effects of size,individual weight and algal concentration

Filtration and ingestion rates of Salpa fusiformis Cuvier were determined while fed Phaeodactylumtricomutum Bohlin at concentrations of 2?64 × 10³ cells·ml^?1. Filtration and ingestion rates increase exponentially with increasing length and body protein. The relations between protein content and body length, and between filtration rate and weight are similar for blastozooids and oozooids. A capture efficiency of the order of 6–32% is calculated: Salpa fusiformis seems to have a low retention efficiency, but its very high filtration rate gives it pride of place amongst filter-feeders. Specific filtration rates are independent of weight; specific ingestion rates are independent of weight for blastozooids, but for oozooids they seem to diminish with increasing weight. The mean daily ration (μg C ingested · μg body C^?1) is 107% for a blastozooid and 117% for an oozooid. Specific filtration rates decrease exponentially as particle concentration rises, as for many other filter-feeders, and the specific ingestion rate follows an Ivlev relation.     

Swallowing motor pattern triggered and modified by sucrose stimulation in the larvae of the silkworm, Bombyx mori

To determine the contribution of sucrose signals to swallowing motor patterns, a series of behavioral, morphological and electrophysiological experiments were carried out in the larvae of the silkworm, Bombyx mori. The larvae ingested a droplet of sucrose solution applied to the mouth. The rate of ingestion was increased for higher sucrose concentrations. The swallowing movements were produced by a cibarial pump system that consisted of a circular compressor and pairs of dilators. The circular compressor was innervated by at least two dorsal motor neurons with the somata in the frontal ganglion. One of these neurons with arborized in both the frontal ganglion and the tritocerebrum of the brain. Both extra- and intracellular recording from the compressor showed that the rhythmic motor patterns were modified by different concentration of sucrose. A higher concentration of sucrose lengthened the duration of a burst or caused more excitatory junction potentials (EJPs) in the compressor, resulting in stronger swallowing contractions. Transection of both frontal connectives deleted the sucrose response, but spontaneous rhythmic motor patterns remained in the compressor, suggesting that the motor rhythm could be generated in the frontal ganglion, and triggered and/or modified by sucrose signals processed through the tritocerebrum of the brain.     

Immunohistological localization of serotonin in the CNS and feeding system of the stable fly Stomoxys calcitrans L. (Diptera: Muscidae)

Serotonin, or 5-hydroxytryptamine (5-HT), plays critical roles as a neurotransmitter and neuromodulator that control or modulate many behaviors in insects, such as feeding. Neurons immunoreactive (IR) to 5-HT were detected in the central nervous system (CNS) of the larval and adult stages of the stable fly, Stomoxys calcitrans, using an immunohistological technique. The location and pattern of the 5-HT IR neurons are described and compared for these two different developmental stages. Anatomical features of the fly feeding system were analyzed in third instar larvae and adult flies using a combination of histological and immunohistological techniques. In third instar larvae, the cibarial dilator muscles were observed within the cibarial pump skeleton and innervated by 5-HT IR neurons in nerves arising from the brain. There were four pairs of nerves arising from the frontal surface of the larval brain that innervate the cibarial pump muscles, pharynx, and muscles controlling the mouth hooks. A strong serotoninergic innervation of the anterior stomatogastric system was observed, which suggests 5-HT may play a role in the coordination of different phases of food ingestion by larvae. Similarly, many 5-HT IR neurons were found in both the brain and the thoracico-abdominal ganglia in the adult, some of which innervate the cibarial pump dilator muscles and the stomatogastric muscles. This is tnhe first report describing neuromuscular structures of the stable fly feeding system. The results reported here suggest 5-HT may play a critical role in feeding behaviors of stable fly larvae and adults.     

Viscometric and potentiometric study of high-methoxyl pectins in the presence of sucrose

The polyelectrolytic behaviour of high-methoxyl pectins (degree of esterification = 72·9) was studied in the presence of sucrose. Potentiometric measurements were carried out on the acidic form by titration with NaOH. Variations of the apparent dissociation constant pK_a as a function of the degree of dissociation α were studied in salt-free solutions on a range of sucrose contents (0–60% w/v). Experimental plots could be described by Lifson-Katchalsky's (LK) treatment up to a 20% sucrose content. The intrinsic apparent dissociation constant pK₀ value was 3·1 ± 0·1 in agreement with previous data on pectinic acid. Above the 20% sucrose content, the LK theory was not appropriate owing to aggregation phenomena of macromolecular chains. Viscometric measurement were performed on the sodium form (α = 1) and for three sucrose contents (0%, 40% and 60%). By carrying out isoionic dilutions, the effect of ionic strength (I_t) on the intrinsic viscosity [η] was studied. The same infinite limit value [η]_∞ was obtained for each sucrose content and linear variations of [η] as a function of $\text{I}{}_{\mathrm{<mtext>t</mtext>}}{}^{\mathrm{<mtext>?1</mtext><mtext>2</mtext>}}$ allowed an estimate of the relative stiffness of the macromolecular chain. This stiffness was slightly increased by sucrose addition.From these data, it could be concluded that a high sucrose content did not significantly change the macromolecular size (for α = 1), as shown by viscometry. However, when α decreased, potentiometric investigations with sucrose showed a higher value of pK_a and a down-curvature of pK_a(α) which could be interpreted by assuming aggregation. These occurred at about 40% sucrose and above which was a much lower concentration than the gelling conditions (about 60%). Therefore aggregation phenomena could be present for relatively low sucrose contents if α < 0·5. It was postulated that interchain associations could occur progressively (from 40 to 60%) and involve aggregate formation with a critical size for gelation.