The Role of Tactile and Chemical Stimuli in the Formation and Maintenance of the Processions of the Social Caterpillar Hylesia lineata (Lepidoptera: Saturniidae)

Colonies of the social caterpillar Hylesia lineata (Lepidoptera: Satumiidae) form long, single-file, head-to-tail processions as they move between their shelters and distant feeding sites. Although investigations of other processionary species have implicated a silk trail in the processionary process, silk plays little or no role in initiating or maintaining processions in H. lineata. Studies we report here implicate both tactile stimuli and a trail pheromone in the establishment and maintenance of processions. Processionaries elicit locomotion in the individual preceding them in line by brushing their heads against prominent sulci that project from the tips of their abdomens. Caterpillars mark their pathways with a pheromone deposited by brushing the ventral surfaces of their last abdominal segments against the substrate. The persistent pheromone is soluble in hexanes and appears to be secreted from glandular setae found on the proximal regions of the anal prolegs and the venter. In Y-choice tests, caterpillars selected newer trails over older trails and stronger trails over weaker trails. They did not distinguish between trials deposited by newly fed caterpillars and those deposited by starved caterpillars. Despite the unidirectional nature of processions, there is no indication that caterpillars can determine from the trail alone the direction in which the procession advanced. The significance of these findings to the foraging ecology of the caterpillars is discussed.     

Trail Marking by the Larva of the Madrone Butterfly Eucheira socialis and the Role of the Trail Pheromone in Communal Foraging Behavior

The larva of the Madrone butterfly Eucheira socialis (Lepidoptera: Pieridae) secretes a trail pheromone from the ventral surface of the posterior tip of its abdomen. Caterpillars mark trails by bringing the secretory site into brief contact with the substrate during a locomotive cycle. Foragers mark most heavily when they move onto new branches and little, if at all, when they move over established trails or when they return to the communal shelter after feeding. The caterpillars make careful comparisons of alternative pathways at choice points and select newer and stronger trails over older and weaker trails. Differential marking of new and established trails during nightly forays, coupled with sensory discrimination of trails by strength and age, leads colonies to abandon old trails in favor of new trails. When applied at a rate as low as 2.5 × 10 ^–10 g/mm, caterpillars followed synthetic trails prepared from 5-cholestane-3-one, a trail pheromone previously reported from the tent caterpillars (Malacosoma spp.). Although both Eucheira and Malacosoma mark with the tip of the abdomen and have near-identical sensitivites to 5-cholestane-3-one, our study shows that Eucheira employs a relatively unsophisticated system of trail-based communication and does not recruit to food. The trail-based communication system of Eucheira appears to represent an early stage in the evolution of cooperative foraging that is derived from, and motivationally linked to, conflict behavior.     

Trail-Based Communication in Social Caterpillars of Eriogaster lanestris (Lepidoptera: Lasiocampidae)

Caterpillars of Eriogaster lanestris (Lepidoptera: Lasiocampidae) mark trails as they move between feeding sites and their communal tent. They prefer new trails over aged ones. Hungry caterpillars prefer trails marked by fed caterpillars returning to the tent. Thus successful foragers direct tentmates to profitable food finds, in a manner similar to that of Malacosoma americanum. E. lanestris readily follows trails prepared from 5-cholestane-3-one, a component of the trail marker of M. americanum, when applied at a rate of 10^–10 g/cm. In choice tests, they preferred more highly concentrated over weaker trails. New trails are always established in groups, and the velocity increases from early to late caterpillars traversing a new trail.     

Spatial and temporal variation in pheromone composition of ant foraging trails

Many social insects use pheromones to communicate and coordinatetheir activities. Investigation of intraspecific differencesin pheromone use is a new area of social insect research. Forexample, interindividual variation in alarm pheromone contenthas been found in physical castes of polymorphic ants. Manyant species use multiple trail pheromones. Here we present novelresearch into trail pheromone variations between behavioralsubcastes of pharaoh ants, Monomorium pharaonis. Monomoriumpharaonis is attracted to trail pheromones found in its poisonglands (monomorines) and Dufour's glands (faranal). We showthat the most abundant monomorines, I (M1) and III (M3), canbe readily detected in pheromone trails. A behaviorally distinctsubcaste known as "pathfinder" foragers can relocate long-livedpheromone trails. Chemical analysis showed that pathfinder foragershad low M3:M1 ratios (mean 3.09 ± 1.53, range 1.03–7.10).Nonpathfinder foragers had significantly greater M3:M1 ratios(38.3 ± 60.0, range 3.54–289). We found that M3:M1ratio did not differ between foragers of different age but wascorrelated with behavioral subcaste at all ages. The relativeabundance of M3:M1 on foraging trails ranged from 3.03–41.3over time during pheromone trail build-up. M3:M1 ratio alsovaried spatially throughout trail networks, being lowest ontrail sections closest to a food source (M3:M1 = 1.9–3.61)and highest near the nest (M3:M1 = 67–267). Our resultsindicate a functional role for differences in pheromone trailcomposition, whereby pathfinder foragers might preferentiallymark sections of pheromone trail networks for future exploration.     

Enhancement of polysaccharides production in <Emphasis Type="Italic">Ganoderma lucidum</Emphasis> by the addition of ethyl acetate extracts from <Emphasis Type="Italic">Eupolyphaga sinensis</Emphasis> and <Emphasis Type="Italic">Catharsius molossus</Emphasis>

To screen stimulators from Chinese medicinal insects for mycelial growth and polysaccharides production of Ganoderma lucidum, G. lucidum was inoculated into the media with and without supplementation of medicinal insect extracts. The ethyl acetate extract of Eupolyphaga sinensis at 55 mg l⁻¹ lead to significant increase in both biomass and intracellular polysaccharides (IPS) concentration from 8.53 ± 0.41 to 14.16 ± 0.43 and 1.28 ± 0.09 to 2.13 ± 0.11 g l⁻¹, respectively. In addition, the ethyl acetate extract of Catharsius molossus at 55 mg l⁻¹ significantly enhanced extracellular polysaccharides (EPS) production; the EPS yield increased from 350.9 ± 14.1 to 475.1 ± 15.3 mg l⁻¹. There were no new components in the two types of polysaccharides obtained by the addition of the insect extracts.     

Trail‐sharing among tropical ants: interspecific use of trail pheromones?

1. Trail‐sharing between different ant species is rare and restricted to a small number of species pairs. Its underlying mechanisms are largely unknown. For trail‐sharing to occur, two factors are required: (i) one or both species must recognise the other species or its pheromone trails and (ii) both species must tolerate each other to a certain extent to allow joint use of the trail. A species that follows another's trails can efficiently exploit the other's information on food sources contained in the pheromone trails. Hence, food competition and thus aggressive interactions between a species following another's trail and the species being followed, seem likely. 2. In the present study, we investigated interspecific trail following and interspecific aggression in trail sharing associations (i) among Polyrhachis ypsilon, Camponotus saundersi, and Dolichoderus cuspidatus, and (ii) among Camponotus rufifemur and Crematogaster modiglianii. We tested whether trail‐sharing species follow each other's pheromone trails, and whether the ants tolerated or attacked their trail‐sharing partners. In both associations, we confronted workers with pheromone trails of their associated species, and, for the former association, measured interspecific aggression among the trail‐sharing species. 3. In our assays, D. cuspidatus and C. rufifemur regularly followed heterospecific pheromone trails of P. ypsilon and C. modiglianii, respectively. However, only few workers of the remaining species followed heterospecific pheromone trails. Thus, shared trails of P. ypsilon and C. saundersi cannot be explained by interspecific trail‐following. 4. Interspecific aggression among P. ypsilon, C. saundersi, and D. cuspidatus was strongly asymmetric, C. saundersi being submissive to the other two. All three species differentiated between heterospecific workers from the same or another site, suggesting habituation to the respective trail‐sharing partners. We therefore hypothesise that differential tolerance by dominant ant species may be mediated by selective habituation towards submissive species and this way determines the assembly of trail‐sharing associations.     

Permeabilization of <Emphasis Type="Italic">Microbacterium oxylans</Emphasis> shifts the conversion of puerarin from puerarin-7-<Emphasis Type="Italic">O</Emphasis>-glucoside to puerarin-7-<Emphasis Type="Italic">O</Emphasis>-fructoside

The main product of the conversion of puerarin by unpermeabilized cells of bacterium Microbacterium oxydans CGMCC 1788 was puerarin-7-O-glucoside (241 ± 31.9 μM). Permeabilization with 40% ethanol could not increase conversion yield, whereas it resulted in change of main product; a previous trace product became a main product (213 ± 48.0 μM) which was identified as a novel puerarin-7-O-fructoside by electrospray ionization time-of-flight MS, ¹³C NMR, ¹H NMR, and GC-MS analysis of sugar composition, and puerarin-7-O-glucoside became a trace product (14.8 ± 5.4 μM). However, the extract from cells of M. oxydans CGMCC 1788 permeabilized with ethanol converted puerarin to form 113.9 ± 27.7 μM puerarin-7-O-glucoside and 187.8 ± 29.5 μM puerarin-7-O-fructoside under the same conditions. When unpermeabilized intact cells were recovered and used repeatedly for the conversion of puerarin, with increase of reuse times, the yield of puerarin-7-O-glucoside gradually decreased, whereas the yield of puerarin-7-O-fructoside increased gradually in the conversion mixture. The main product of the conversion of puerarin by the tenth recycled unpremerbilized cells was puerarin-7-O-fructoside (288.4 ± 24.0 μM). Therefore, the change of permeability of cell membrane of bacterium M. oxydans CGMCC 1788 contributed to the change of conversion of the product’s composition.     

Habitat use,feeding and territorial behavior of a Brazilian endemic damselfish <Emphasis Type="Italic">Stegastes rocasensis</Emphasis> (Actinopterygii: Pomacentridae)

This paper presents the results of the first field study of the Brazilian endemic damselfish Stegastes rocasensis. Underwater data were collected at the Fernando de Noronha Archipelago (NE Brazil), where four sites were visually assayed for local territorial and feeding behaviors, as well as habitat use. S. rocasensis preferentially fed on turf and macroalgae, and during the afternoon period. Its territory size ranged from 0.26 to 13.86 m² (1.61 ± 0.23 m²). Our findings suggest that adults preferably inhabit shallow, turf-rich areas, while sub-adults and juveniles occupy deeper areas with higher percent cover of invertebrates and hard corals, respectively. With regards to agonistic behaviors, pursuit of intruders was the most frequent reaction recorded (p < 0.001); the highest level of aggression was elicited by territorial herbivores (p < 0.001). Nevertheless, analysis of attacks elicited by different fish species within S. rocasensis’ territory showed that more attacks were launched on conspecifics than on other fish species.     

Mitochondrial DNA reveals low population differentiation in elongate loach, <Emphasis Type="Italic">Leptobotia elongata</Emphasis> (Bleeker): implications for conservation

Elongate loach (Leptobotia elongata (Bleeker)), an endemic fish species to China, is a famous ornamental freshwater fish. Here, a comparative study of mtDNA control region (D-loop) (835 bp) sequences was performed to analyze its wild population structure and evaluate the genetic diversity for 110 individuals from five locations in the upper reaches of the Yangtze River, China. A total of 49 polymorphic sites and 45 haplotypes yielded high haplotype diversity (h = 0.952), but low nucleotide diversity (π = 0.00454) as that of many fish species. Sequence divergences between haplotypes ranged from 0.0033 ± 0.0011 to 0.0050 ± 0.0012 in intra-groups, and from 0.0037 ± 0.0.0011 to 0.0050 ± 0.0012 between groups. Significant values of Tajima’s D (−1.86383, P < 0.01) and Fu’s F _S (−25.93, P < 0.01), together with uni-modal mismatch distribution, indicated a recent genetic bottleneck or population expansion of the species. Analysis of molecular variance (AMOVA) indicated a small amount of differentiation among groups (1.7%); most of the total variation occurred within groups (98.3%). Also, there was no significant population structure (F _ST = 0.017, P > 0.05), and estimates of gene flows among groups were extremely high (Nm = 28.88), suggesting low genetic divergence between populations in the species. The lack of genetic differentiation among groups is most likely due to the combined gene flow from the downstream movement of eggs and larvae with currents and the upstream or downstream migration of adults throughout the distribution. These groups of L. elongata distributed in upper reaches of the Yangtze River should be considered as a single management unit.     

Trail following and stowaway behaviour of the myrmecophilous staphylinid beetle,Homoeusa acuminata,during foraging trips of its hostLasius fuliginosus (Hymenoptera: Formicidae)

Summary The behaviour of adultHomoeusa acuminata on trails of its hostLasius fuliginosus was investigated both in the field and in the laboratory. The beetles were active from May to September, accurately following the foraging trails of their hosts up to 20 metres from the nest. Most of the time, they were ignored by the ants, but if attacked they raised their abdomen as a possible appeasement or defensive behaviour. On trails the beetles most probably act as food robbers, feeding on prey collected by ants. The following method, called stowaway behaviour, was used by the beetles: when a beetle encountered an ant carrying a prey back to the nest it jumped on the prey, probably feeding on it while being transported.Laboratory experiments on circular artificial trails demonstrated thatH. acuminata follows a water extract of hindguts of the ants, the source of the trail pheromone. Both beetles and ants responded to an artificial trail of 0.03 hindgut equivalent per cm, but the mean distance followed by the beetles was about twelve times higher than that covered by the ants themselves. In contrast, experiments with solutions of the six fatty acids reported as the active components of the trail pheromone showed that the beetles did not respond at all, and that the ants only respond to the fatty acids at a very high concentration.     

Host-ant trail following by myrmecophilous larvae of Liphyrinae (Lepidoptera,Lycaenidae)

In this study we report a case of ant-trail following by lycaenid caterpillars. Euliphyra mirifica and E. leucyana caterpillars are involved in a commensal association with the weaver ant Oecophylla longinoda. The host nests are made with leaves which over the course of time dry out or are broken open by storms, forcing the ants to migrate and build a new nest elsewhere. Euliphyra caterpillars are stimulated by recruitment behaviour which triggers the migration of their host. They then follow the host trails leading to the new nesting site. Laboratory experiments showed that these caterpillars are able to follow host trails under varied conditions: (1) fresh trails actually used by workers, (2) fresh trails in the absence of workers, (3) heterocolonial, 2-month-old trails, and (4) fresh trails washed with water (to simulate the effect of tropical rains). They can also bridge trail gaps of more than 1 cm. Under natural conditions, the trails are frequently situated along thin twigs. The forward progress of the ants in such a situation is not impeded by the presence of large Euliphyra larvae. Workers just climb over the caterpillars, even on larger trails where there is enough room to pass alongside them. This suggests that an allomone is secreted on the dorsal part of the caterpillars. When crawling along heterocolonial trails, the caterpillars are not attacked, even if about 21% of the workers from the new colony spread their mandibles when encountering them. They are then adopted and are admitted to the nest of the new host colony of O. longinoda.     

Effect of Different Iodine Nutrition on Cerebellum Pcp-2 in Rat Offspring During Lactation

The thyroid functions of breastfed infants, as well as (indirectly) the development of their central nervous system, are dependent on the iodine status of the lactating mother. Purkinje cell protein-2 is a cell-specific marker of the cerebellum Purkinje cell and is a suitable indicator for observing the postnatal development of the cerebellum after birth. We measured the Purkinje cell protein-2 mRNA and protein levels in the rat cerebellum in the critical postnatal (14 days after birth) and maturation periods (28 days after birth) to determine the effect of different nutritional iodine levels on cerebellum growth in the offspring during lactation. We found that severe iodine deficiency resulted in thyroid dysfunction in lactating rats and their offspring on both 14 and 28 days, showing maternal total T₄ 16.7 ± 12.0 vs 36.4 ± 15.0, P < 0.05 (14 days) and 22.6 ± 18.7 vs 53.4 ± 9.4, P < 0.01 (28 days), and neonatal total T₄ 10.6 ± 2.3 vs 16.4 ± 4.7, P < 0.01(14 days) and 12.8 ± 2.9 vs 16.7 ± 3.4, P < 0.05 (28 days), respectively. The Purkinje cell protein-2 mRNA and its protein levels in offspring rats were significantly reduced that showed Purkinje cell protein-2 mRNA 1.12 ± 0.04 vs 2.25 ± 0.53, P < 0.05 (14 days) and 1.74 ± 0.94 vs 8.69 ± 2.71, P < 0.01 (28 days). However, mild iodine deficiency and excessive iodine maintained almost normal thyroid function in maternal and neonatal rats and normal Purkinje cell protein-2 mRNA and protein levels in offspring’s cerebellum. We conclude that severe iodine deficiency could significantly reduce Purkinje cell protein-2 mRNA and its protein levels, indicating that the cerebellum development was retarded, but mild iodine deficiency and excessive iodine could maintain them at an approximately normal level by the mother’s and offspring’s compensations, especially by the mother’s mammary glands.     

Characteristics of Night Sleeping Trees of Proboscis Monkeys (<Emphasis Type="BoldItalic">Nasalis larvatus</Emphasis>) in Sabah,Malaysia

Primates spend about half of their lives at sleeping sites, and their choice of sleeping sites may affect individual survival. We identified a total of 88 trees used by proboscis monkeys (Nasalis larvatus) as night sleeping sites on 16 nights from June to September 2008 in riverine, mangrove, and mixed mangrove–riverine forests along the Garama River, a tributary of the Klias River, in the west of Sabah, Malaysia. We recorded 11 variables for each tree, including the species, physical structure, distance from the riverbank, and connectivity with surrounding trees. We compared sleeping trees with 114 trees with ≥30 cm girth at breast height (GBH) located ≤50 m of the riverbank in 8 botanical plots (total 1 ha). Trees in the plots represented the general vegetation patterns of the study area. Choice of sleeping trees did not depend on the tree species. Although sleeping trees included trees ≤46 m from the river, those closer to riverbanks (5–35 m, n = 76) were more likely to be used as sleeping sites. Compared to the available trees, sleeping trees had larger trunks (mean±SD = 143.6 ± 56.9 cm GBH), and were taller (mean±SD = 34.3 ± 8.1 m), with greater number (median = 6; range = 12) and larger (mean±SD = 24.1 ± 15.2 cm circumference) main branches. They were also located near to other trees, with overlapping branches, creating good arboreal connectivity. Choice of sleeping trees by proboscis monkeys is likely to be related to risks of predation and injury from falling, as well as ease of social interaction and efficiency of locomotion.     

Tracking the Ovarian Cycle in Black-and-Gold Howlers (<Emphasis Type="Italic">Alouatta caraya</Emphasis>) by Measuring Fecal Steroids and Observing Vaginal Bleeding

A better understanding of a species’ reproductive physiology can help conservation programs to manage primates in the wild and develop assisted reproductive technologies in captivity. We investigated whether measurements of fecal progestin and estrogen metabolites obtained by a radioimmunoassay could be used to monitor the ovarian cycle of Alouatta caraya. We also compared the occurrence of vaginal bleeding with the hormone profiles. We collected fecal samples from 3 adult and 1 subadult captive female over 5 mo and performed vaginal cytology for the adults. The interval between fecal progestin surges in the adult females was 19.11 ± 2.14 d (n = 18 cycles). Fecal progestin concentrations remained at basal values for 9.83 ± 2.21 d (n = 18) and rose to elevated values for 9.47 ± 0.72 d (n = 19). The subadult female showed basal levels of fecal estrogen and progestin concentrations throughout the study, suggesting that our hormone measurements are valid to monitor the ovarian cycle. Bleeding periods coincided with basal levels of fecal estrogens and progestin at intervals of 19.8 ± 0.9 d and lasted for 4.1 ± 1.0 d. Although we obtained these data from only 3 individuals, the results indicate that this species likely has a menstrual-type ovarian cycle. These data provide the first endocrine profile for the Alouatta caraya ovarian cycle and are similar to results obtained for other howler species. This similarity is important for comparative studies of howlers, allowing for a better understanding of their reproductive physiology and contributing to a critical information base for managing Alouatta species.     

Gastrointestinal passage time of seeds ingested by captive Japanese martens <Emphasis Type="Italic">Martes melampus</Emphasis>

The time it takes for ingested seeds to pass through the gut of animals is an important aspect of endozoochorous seed dispersal because it influences seed dispersal distance. Variations in the physical characteristics of seeds, such as their weight, volume, and specific gravity, can affect their movement through the gastrointestinal system of a given animal. We conducted feeding experiments with captive Japanese martens, Martes melampus (n = 4), at Toyama Municipal Family Park Zoo, central Japan to examine the effects of the physical characteristics of seeds on their passage times. The mean (±SD) transit time, mean retention time, and time of last appearance of four different types of commercial seeds were 2.6 ± 0.3 h (range 0.6–5.4), 9.7 ± 1.1 h (3.8–17.3), and 23.8 ± 3.1 h (12.2–51.8), respectively. All of these values are greater than those found during previous experiments conducted with mustelids. Similar to previous studies, however, none of these passage time variables was correlated with the physical characteristics of seeds. Our results thus indicate that martens disperse seeds of different plant species, whose size, volume, and specific gravity all fall within the range of those used in the present study, from parent plants at similar distances.     

Stingless bees (<Emphasis Type="Italic">Scaptotrigona pectoralis</Emphasis>) learn foreign trail pheromones and use them to find food

Foragers of several species of stingless bees (Hymenoptera, Apidae and Meliponini) deposit pheromone marks in the vegetation to guide nestmates to new food sources. These pheromones are produced in the labial glands and are nest and species specific. Thus, an important question is how recruited foragers recognize their nestmates’ pheromone in the field. We tested whether naïve workers learn a specific trail pheromone composition while being recruited by nestmates inside the hive in the species Scaptotrigona pectoralis. We installed artificial scent trails branching off from trails deposited by recruiting foragers and registered whether newly recruited bees follow these trails. The artificial trails were baited with trail pheromones of workers collected from foreign S. pectoralis colonies. When the same foreign trail pheromone was presented inside the experimental hives while recruitment took place a significant higher number of bees followed the artificial trails than in experiments without intranidal presentation. Our results demonstrate that recruits of S. pectoralis can learn the composition of specific trail pheromone bouquets inside the nest and subsequently follow this pheromone in the field. We, therefore, suggest that trail pheromone recognition in S. pectoralis is based on a flexible learning process rather than being a genetically fixed behaviour.     

RETRACTED ARTICLE: Archaeal <Emphasis Type="Italic">amoA</Emphasis> Genes Outnumber Bacterial <Emphasis Type="Italic">amoA</Emphasis> Genes in Municipal Wastewater Treatment Plants in Bangkok

The contribution of ammonia-oxidizing archaea (AOA) to nitrogen removal in wastewater treatment plants (WWTPs) remains unknown. This study investigated the abundance of archaeal (AOA) and bacterial (ammonia-oxidizing bacteria (AOB)) amoA genes in eight of Bangkok’s municipal WWTPs. AOA amoA genes (3.28 × 10⁷ ± 1.74 × 10⁷–2.23 × 10¹¹ ± 1.92 × 10¹¹ copies l⁻¹ sludge) outnumbered AOB amoA genes in most of the WWTPs even though the plants’ treatment processes, influent and effluent characteristics, removal efficiencies, and operation varied. An estimation of the ammonia-oxidizing activity of AOA and AOB suggests that AOA involved in autotrophic ammonia oxidation in the WWTPs. Statistical analysis shows that the numbers of AOA amoA genes correlated negatively to the ammonium levels in effluent wastewater, while no correlation was found between the AOA amoA gene numbers and the oxygen concentrations in aeration tanks. An analysis of the AOB sequences shows that AOB found in the WWTPs limited to only two AOB clusters which exhibit high or moderate affinity to ammonia. In contrast to AOB, AOA sequences of various clusters were retrieved, and they were previously recovered from a variety of environments, such as thermal and marine environments.     

Comparison of metabolite levels in callus of <Emphasis Type="Italic">Tecoma stans</Emphasis> (L.) Juss. ex Kunth. cultured in photoperiod and darkness

Tecoma stans is a tropical plant from the Americas. Antioxidant activity and both phenolic compound and flavonoid total content were determined for callus tissue of T. stans cultured in either a set photoperiod or in darkness. Callus lines from three explant types (hypocotyls, stem, and leaf) were established on B5 culture medium supplemented with 0.5 μM 2,4-D and 5.0 μM kinetin. While leaf-derived callus grew slower under a 16-h photoperiod (specific growth rate, μ = 0.179 d⁻¹, t _D = 3.9 d) than in darkness (μ = 0.236 d⁻¹, t _D = 2.9 d), it accumulated the highest amount (p < 0.05) of both phenolics (86.6 ± 0.01 mg gallic acid equivalents/g) and flavonoids (339.6 ± 0.06 mg catechin equivalents/g). Similarly, antioxidant activity was significantly higher (p < 0.05) when callus was cultured in period light than when grown in extended darkness. Antioxidant activity measured with a 2,20-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS)-based assay was 350.5 ± 15.8 mmol Trolox/g extract for callus cultured under a defined photoperiod compared to 129.1 ± 7.5 mmol Trolox/g extract from callus cultured in darkness. Content of phenolic compounds and flavonoids was in agreement with a better antioxidant power (EC₅₀ = 450 μg extract/mg 1,1-diphenyl-2-picrylhydrazyl) and antiradical efficiency. Results of the present study show that calli of T. stans are a source of compounds with antioxidant activity that is favored by culture under a set photoperiod.     

Pathways for K<Superscript>+</Superscript> Efflux in Isolated Surface and Crypt Colonic Cells. Activation by Calcium

K⁺-conductive pathways were evaluated in isolated surface and crypt colonic cells, by measuring ⁸⁶Rb efflux. In crypt cells, basal K⁺ efflux (rate constant: 0.24 ± 0.044 min⁻¹, span: 24 ± 1.3%) was inhibited by 30 mM TEA and 5 mM Ba²⁺ in an additive way, suggesting the existence of two different conductive pathways. Basal efflux was insensitive to apamin, iberiotoxin, charybdotoxin and clotrimazole. Ionomycin (5 μM) stimulated K⁺ efflux, increasing the rate constant to 0.65 ± 0.007 min⁻¹ and the span to 83 ± 3.2%. Ionomycin-induced K⁺ efflux was inhibited by clotrimazole (IC₅₀ of 25 ± 0.4 μM) and charybdotoxin (IC₅₀ of 65 ± 5.0 nM) and was insensitive to TEA, Ba²⁺, apamin and iberiotoxin, suggesting that this conductive pathway is related to the Ca²⁺-activated intermediate-conductance K⁺ channels (IK_ca). Absence of extracellular Ca²⁺ did neither affect basal nor ionomycin-induced K⁺ efflux. However, intracellular Ca²⁺ depletion totally inhibited the ionomycin-induced K⁺ efflux, indicating that the activation of these K⁺ channels mainly depends on intracellular calcium liberation. K⁺ efflux was stimulated by intracellular Ca²⁺ with an EC₅₀ of 1.1 ± 0.04 μM. In surface cells, K⁺ efflux (rate constant: 0.17 ± 0.027 min⁻¹; span: 25 ± 3.4%) was insensitive to TEA and Ba²⁺. However, ionomycin induced K⁺ efflux with characteristics identical to that observed in crypt cells. In conclusion, both surface and crypt cells present IK_Ca channels but only crypt cells have TEA- and Ba²⁺-sensitive conductive pathways, which would determine their participation in colonic K⁺ secretion.     

Some reproductive characteristics of endangered Caspian Lamprey (<Emphasis Type="Italic">Caspiomyzon wagneri</Emphasis> Kessler, 1870) in the Shirud River southern Caspian Sea,Iran

Migration and reproduction of the Caspian Lamprey, Caspiomyzon wagneri, in the Shirud River were investigated during late-March to early-May at water temperatures ranging from 11 to 21.25°C. We examined the effect of water temperature on timing of spawning migrations. There was a significant negative relationship between temperature and intensive migration of Caspian Lamprey (p < 0.05). The most intensive migration of lampreys was at night (21:00–3:00 h) and when the water temperatures averaged 16°C (34.43%). The overall sex ratio (male to female) was 1.07 to 1. The individual absolute fecundity was 31 ‘758–51’ 198 eggs (mean±SD—41,924 ± 5,382). The egg diameter was 0.780–1.151 (0.92 ± 0.081) mm. The individual relative fecundity varies from 80.3 to 148.1 (107.2 ± 15.1) eggs per 1 mm of length and from 260.8 to 677.4 (397.6 ± 93) eggs per 1 g of weight. The gonadosomatic index (GSI) of females was 5.83–31.44 (11.22 ± 4.30).