Possible involvement of the glycocalyx in the ichthyotoxicity of Chattonella marina (Raphidophyceae): immunological approach using antiserum against cell surface structures of the flagellate

Although the ichthyotoxic mechanism of Chattonella marina is still unknown, several lines of evidence suggest that the reactive oxygen species (ROS), such as superoxide anion (O₂^-), hydrogen peroxide (H₂O₂)_, and hydroxyl radical (·OH), produced by C. marina are involved in the mortality of fish exposed to this flagellate. Recently, we found that the cell-free supernatant prepared from C. marina, which is considered to contain the glycocalyx, showed NADPH-dependent O₂^- generation. In this study, we prepared antiserum against the crude glycocalyx of C. marina. Using indirect immunofluorescence, it was confirmed that the antiserum specifically reacted with C. marina cells. In addition to C. marina, the antiserum also reacted with other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica, whereas no reactivity was observed against six other flagellate species tested. These results suggest that raphidophycean flagellates have common epitopes recognized by the antiserum. Interestingly, immunohistochemical analysis of paraformaldehyde-fixed gill lamellae from yellowtail exposed to C. marina revealed that the antiserum stained the surface of gill lamellae, while no such staining pattern was observed in control gill lamellae. These results suggest that the glycocalyx may be discharged when C. marina cells are inhaled into the fishes' mouths and then come into contact with the gill surface. Based on the present results, together with our previous findings, we propose that continuous accumulation of the discharged glycocalyx on the gill surface occurs during C. marina exposure, which may be responsible for the ROS-mediated severe gill tissue damage leading to fish death.     

Regulation of ovary activation in worker honey-bees (Apis mellifera): larval signal production and adult response thresholds differ between anarchistic and wild-type bees

One-day-old anarchistic (selected for successful worker reproduction) and wild-type honey-bee workers were introduced into queenright colonies of honey-bees of two treatments. In treatment 1, all eggs and larvae were offspring of queens from an anarchistic line. In treatment 2, all eggs and larvae were offspring of wild-type queens. In both treatments, adult workers were wild type. This experimental arrangement was used to test the importance of larval genotype on ovary activation in young adult workers. After 12 days, the introduced bees were dissected to determine the frequency of ovary activation. In those colonies provided with wild-type brood, 0% of introduced wild-type bees and 16% of anarchistic bees had activated ovaries. In those colonies provided with anarchistic brood, 13% of introduced wild-type bees and 41% of anarchistic bees had activated ovaries. These results strongly support the hypothesis that selection for high levels of worker reproduction in anarchistic stocks has reduced the amount or composition of brood pheromones produced by larvae that normally signal workers to refrain from reproduction. They also suggest that anarchistic workers have a higher threshold for these signals than wild-type bees.     

Anti-sense expression of putrescine N-methyltransferase confirms defensive role of nicotine in Nicotiana sylvestris against Manduca sexta

Summary. Several lines of evidence support the defensive function of nicotine production in the Nicotiana genus against a range of herbivores, but the evidence is largely correlative. To suppress nicotine production in planta and to test its defensive function, we expressed DNA of putrescine N-methyl transferase in an anti-sense orientation (AS-PMT) in N. sylvestris and fed leaf material from two lines of transformed and wild type plants to Manduca sexta larvae. Larvae consumed more leaf area and gained more mass on the foliage of plants with low PMT expression and low nicotine levels as compared to plants with high PMT expression and high nicotine levels and wild type plants. Overall, larval consumption and performance were negatively correlated with constitutive nicotine levels. We conclude that nicotine decreases the palatability of N. sylvestris leaves to the nicotine-resistant M. sexta larvae.     

Role of Genetic Background in the Success of Reintroduced Peregrine Falcons

Abstract: Peregrine Falcons (    Falco peregrinus ) of seven subspecies from four continents were bred in captivity, and approximately 1173 of their progeny were released in the midwestern United States and adjacent regions of Ontario and Manitoba in an attempt to replace the original population that was extirpated by chlorinated hydrocarbon poisoning in the 1950s. We analyzed the success of individuals of the different subspecies introduced to the Midwest. Five of the seven subspecies released have contributed to the current breeding population. Subspecies of breeding Peregrine Falcons were equally represented when breeding birds of high productivity were compared with less prolific breeders. The subspecific makeup of the breeding population did not differ significantly from that of the released population, suggesting that adaptability in this species was sufficient to override genetic differences between subspecies. Peregrines of widely different genetic stocks have thrived after release, making substantial genetic contributions to the new population.     

Simultaneous hermaphroditism in the marine shrimp Lysmata wurdemanni (Caridea: Hippolytidae): an undescribed sexual system in the decapod Crustacea

Simultaneous hermaphroditism with outcrossing, a previously unconfirmed sexual system in decapod crustaceans, is documented in the shrimp Lysmata wurdemanni (Gibbes), using time-lapse video observations on mating in pairs of “female-phase” (FP) individuals. Copulations between FPs resulted in successful spawning and development of embryos. However, female-phase hermaphrodites maintained in isolation were unable to self-fertilize spawned eggs. All smaller individuals possessed characters typical of caridean males, including male gonopores, appendices masculinae, ejaculatory ducts, and cincinnuli on the first pleopods. However, the gonad of these male-phase (MP) individuals was an ovotestis with an undeveloped ovarian portion. FPs, which spawn eggs and incubate embryos, also had male gonopores and an ovotestis terminating in ejaculatory ducts containing sperm. In FPs, male pleopod characters were absent or reduced, and a female incubatory character, expanded pleopod flanges, showed a greater relative size. Smaller size classes of the population were composed only of MPs while larger size classes consisted of both MPs and FPs. “Transitionals,” individuals with MP characters but an ovotestis containing vitellogenic oocytes, were rare and overlapped completely in size with FP and larger MP individuals. It is suggested that not all MPs transform into FP hermaphrodites. Social interactions may mediate MP change into FP hermaphrodites in L. wurdemanni. The social system of L. wurdemanni and other Lysmata spp. may determine the exact form of the sexual system in these species. Received: 26 October 1997 / Accepted: 21 April 1998     

Lectin-induced enhancement of superoxide anion production by red tide phytoplankton

Chattonella marina, a raphidophycean flagellate, is a highly toxic red tide phytoplankton which causes severe damage to fish farming. Recent studies demonstrated that Chattonella spp. continuously release superoxide anions (O₂ ⁻) while they are living. Heterosigma akashiwo, another raphidophycean flagellate, also produces O₂ ⁻. In the present study, we found that lectins such as concanavalin A (Con A), wheat germ agglutinin (WGA), and castor bean hemagglutinin (CBH) stimulated  C. marina and H. akashiwo to generate enhanced amounts of O₂ ⁻ in a concentration-dependent manner. The lectin-specific sugars potently inhibited the lectin-induced increase of O₂ ⁻ production, suggesting that the effects of lectins are mediated mainly through the interaction of these lectins with carbohydrate moiety present on the flagellate cell surface. In contrast to the potent ability of native Con A (tetravalent), succinylated Con A (divalent) showed only a slight stimulative effect on these flagellates. O₂ ⁻ production was totally inhibited by treatment with proteinase K for 30 min, without affecting the viabilities of flagellates. These results suggest that cell-surface redox enzymes may be involved in O₂ ⁻ production, and such enzymes are responsible for the lectin-stimulation. Received: 21 August 1997 / Accepted: 8 January 1998     

Paternity and paternal care in the polygynandrous Smith's longspur

In species where females copulate with more than one male during a single breeding attempt, males risk investing in offspring that are not their own. In the polygynandrous Smith's longspur (Calcarius pictus), females copulate sequentially with one to three males for each clutch of eggs and most of these males later assist in feeding the young. Using multilocus DNA profiling, we determined that there was mixed paternity in >75% of broods (n=31) but that few offspring (<1% of 114 nestlings) were sired by males outside the polygynandrous group. Male feeding rate increased significantly with the number of young sired, with males siring four nestlings feeding the brood at double the frequency of males siring only a single nestling. However, male Smith's longspurs appear to show a graded adjustment of paternal care in response to paternity only when other males are available to compensate for reduced care: feeding rate did not vary in relation to paternity when only one male provisioned young at the nest. There was no evidence that males could recognise their own offspring within a brood and feed them preferentially. The number of offspring sired by each male was significantly correlated with the number of days spent copulating with the attending female: on average, a male sired one offspring for every 2 days of copulatory access. If males use their access to females to estimate paternity (and thereby decide on their subsequent level of parental investment), a positive relationship is expected between the amount of female access and the subsequent feeding rate to the nestlings. Nonetheless, male feeding effort was only weakly correlated with female access and more study is needed to determine how males estimate their paternity in a brood. Received: 1 June 1997 / Accepted after revision: 1 April 1998     

Identification and evaluation of hydrogeochemical processes on river Cooum, South India

The salient features of the river water chemistry and the seasonal variations on the individual chemical parameter were evaluated and characterized. The order of abundance of the ions in the water is determined for both seasons. The contribution of chemical weathering to the water chemistry has been determined using the (Ca²⁺ + Mg²⁺)/(Na⁺ + K⁺) ratio, ternary, and Gibb’s diagrams. The results show that the chemical composition of river water during premonsoon is controlled mainly by evaporation–crystallization, while in the postmonsoon, the rock–water interaction dominates. The unique characteristic of the river water is the linear relationship among the principal ions. Hydrochemical characteristics of ions in the water were studied using 1:1 equiline diagrams. The nature of the water samples was determined using the piper diagram. The influence of trace metals on the chemical composition and the quality of the river water in the study area has been assessed using Wilcox and US Salinity Laboratory diagrams.