Effects of the epibiotic heterotrich ciliate Mirofolliculina limnoriae and of moulting on faecal pellet production by the wood-boring isopods, Limnoria tripunctata and Limnoria quadripunctata

The heterotrich ciliate Mirofolliculina limnoriae occurs in an obligate association with wood-boring species of the isopod Limnoria. It attaches to the dorsal surface of the limnoriid pleotelson. Folliculinid-bearing populations of Limnoria tripunctata from the Ria Formosa, Portugal, and of Limnoria quadripunctata from Portsmouth, UK, were sampled. Up to four folliculinids were found on one individual of L. quadripunctata and up to twelve on L. tripunctata. There was no evidence that folliculinid load increased with host size. The nature of the epibiont/basibiont relationship was examined by observing animals with and without epibionts, kept individually with small sticks of wood in cell culture wells over periods of up to 15 days. Faecal pellet production and moulting were recorded. In both host species, feeding rate as measured by faecal pellet production was significantly lower in animals bearing folliculinids. Feeding rate also diminished with the onset of moulting, but returned to the original rate within a week after moulting. The estimated intermoult period for the population of L. tripunctata examined was about 25 days and for L. quadripunctata was about 32 days. However, the proportion of individuals moulting over the period of observation declined with increasing size in L. tripunctata, suggesting increasing length of intermoult period with age. As the host faces toward the blind end of its tunnel and ventilates the burrow with its pleopods, the folliculinids are well placed to intercept particulate food in the incoming water stream. There are no clear advantages for the host in the association, but the suppression of feeding and the potential of the folliculinids to hinder swimming during dispersal migrations indicate that the association should be viewed as ectoparasitism rather than mutualism or commensalism.     

Biogeography of Wood-Boring Crustaceans (Isopoda: Limnoriidae) Established in European Coastal Waters

Marine wood-borers of the Limnoriidae cause great destruction to wooden structures exposed in the marine environment. In this study we collated occurrence data obtained from field surveys, spanning over a period of 10 years, and from an extensive literature review. We aimed to determine which wood-boring limnoriid species are established in European coastal waters; to map their past and recent distribution in Europe in order to infer species range extension or contraction; to determine species environmental requirements using climatic envelopes. Of the six species of wood-boring Limnoria previously reported occurring in Europe, only Limnoria lignorum, L. quadripunctata and L. tripunctata are established in European coastal waters. L. carinata and L. tuberculata have uncertain established status, whereas L. borealis is not established in European waters. The species with the widest distribution in Europe is Limnoria lignorum, which is also the most tolerant species to a range of salinities. L. quadripunctata and L. tripunctata appear to be stenohaline. However, the present study shows that both L. quadripunctata and L. tripunctata are more widespread in Europe than previous reports suggested. Both species have been found occurring in Europe since they were described, and their increased distribution is probably the results of a range expansion. On the other hand L. lignorum appears to be retreating poleward with ocean warming. In certain areas (e.g. southern England, and southern Portugal), limnoriids appear to be very abundant and their activity is rivalling that of teredinids. Therefore, it is important to monitor the distribution and destructive activity of these organisms in Europe.     

The effect of relative humidity on photoresponse in sympatric species of Drosophila: Long-term exposure to desiccating environments

The photoresponse of isofemale strains of the sympatric species Drosophila immigrans and Drosophila tripunctata were compared at relative humidities of 95, 50 and 5% during an 8-h period. The onset of a change in photobehaviour due to the stress of desiccation was calculated for each strain for the three relative humidities. The two species could be distinguished on the basis of their light responses under the three desiccating conditions. With increased desiccation there was an increased positive response to light after the behavioural shift. The effect was greater and more uniform in D. tripunctata, which is a fungus feeder, than in D. immigrans, possibly because D. tripunctata occupies a narrower niche than D. immigran. The species were observed to have higher activity levels as they desiccated. It is postulated that the increased positive light responses with increased relative humidity are attributable to both desiccation and starvation. The time of onset of the shift in photobehaviour is dependent on the relative humidity.     

The larva of Oecetis tripunctata (Fabricius, 1793) (Trichoptera,Leptoceridae)

Oecetis tripunctata is a widely distributed leptocerid in Europe, ranging from the Iberian and Apennine peninsulas and the Central and Western European highlands to the plains of Eastern Europe. The long, single-bladed mandibles are indicative for a predacious lifestyle. This paper describes the previously unknown larva of Oecetis tripunctata. Information on the morphology of the 5th larval instar is given, and the most important diagnostic features are illustrated. A synoptic key for the European species of Oecetis is also provided. In the context of existing identification keys the larva of Oecetis tripunctata keys together with Oecetis intima and Oecetis notata. Oecetis tripunctata is separated from the other two species by the fact that a double row of long setal fringes is lacking at the hind tibiae and that several long setae are present on the protrochantinus.     

Divergence in the neural control of oviposition in Drosophila

Of several species of Drosophila tested, only D. melanogaster was capable of ovipositing after decapitation. Two other species (D. tripunctata and D. pseudoobscura) were also capable of reflex oviposition during the operation but D. virilis and D. palustris were not. This divergence of neural control mechanisms suggests the existence of at least two alternate circuits for the control of insect oviposition.     

Arsenic concentration in the urine of patients with Blackfoot disease and Bowen’s disease

Extensive epidemiological study implicates that high arsenic content in artesian well water is the causal factor responsible for Blackfoot disease. We determine the arsenic concentration in urine samples of patients with Blackfoot and Bowen’s diseases and examine whether there exists any discrepancy of urinary arsenic concentrations among patients and the normal population. The analyses were made by hydride atomic absorption spectrophotometry (AAS) and the analytical reliability of the method was checked with a standard urine sample (ORTHO Bi-Level Urine Metal Control). The results show that the mean urinary arsenic concentration in 100 healthy adults is 63.4±29.7 μg/L, and those means for 23 and 11 patients with Blackfoot disease and Bowen’s disease are 75.7±39.1 μg/L (P vs controls >0.05) and 201±58 μg/L (P vs controls <0.001), respectively. From the analytical results obtained, we cannot conclude that urinary arsenic is associated with Blackfoot disease, as was disclosed from the epidemiological studies. However, urinary arsenic concentrations are possibly very closely associated with Bowen’s disease.     

Historical patterns of niche dynamics in Neotropical species of the <Emphasis Type="Italic">Drosophila</Emphasis> subgenus (Drosophilidae,Diptera)

Niche conservatism (NC) presence is a controversial question in evolutionary ecology. In Drosophila, little is known about which is the preponderant evolutionary pattern, since the adaptive radiation hypothesis first proposed by Throckmorton assumed niche divergence (ND) according to a niche occupancy scenario. Nevertheless, this hypothesis has not yet been straightforwardly tested. Our aim here was to test the role of NC patterns across evolution of American drosophilids belonging to the tripunctata and virilis-repleta lineages of the Drosophila subgenus, through measures of geographical, abiotic and biotic niche overlap and evaluations regarding the presence of phylogenetic signal or niche identity. We recovered phylogenetic signal attributable to phylogenetic niche conservatism when all species were analyzed together, but not in more restricted groups. Identity tests showed that niche equivalency was seldom rejected for the tripunctata lineage species. So, in general, neither the results for the Drosophila subgenus nor those for the tripunctata lineage support the hypothesis of an adaptive radiation. Notwithstanding, there were also several isolated cases supporting a scenario of ND, and ecological speciation was evident in some of the evaluated sister species pairs.     

Molecular phylogeny of the Drosophila tripunctata and closely related species groups (Diptera: Drosophilidae)

We suggest a new phylogenetic hypothesis for the tripunctata radiation based on sequences of mitochondrial genes. Phylogenetic trees were reconstructed by parsimony, maximum likelihood and Bayesian methods. We performed tests for hypotheses of monophyly for taxonomic groups and other specific hypotheses. Results reject the monophyly for the tripunctata group whereas monophyly is not rejected for the tripunctata radiation and other specific groups within the radiation. Although most of the basal nodes were unresolved we were able to identify four clusters within the tripunctata radiation. These results suggest the collection of additional data before a proper taxonomic revision could be proposed.     

Inter‐ and intraspecific variation in mycotoxin tolerance: A study of four Drosophila species

Many mycophagous Drosophila species have adapted to tolerate high concentrations of mycotoxins, an ability not reported in any other eukaryotes. Although an association between mycophagy and mycotoxin tolerance has been established in many Drosophila species, the genetic mechanisms of the tolerance are unknown. This study presents the inter‐ and intraspecific variation in the mycotoxin tolerance trait. We studied the mycotoxin tolerance in four Drosophila species from four separate clades within the immigrans‐tripunctata radiation from two distinct locations. The effect of mycotoxin treatment on 20 isofemale lines per species was studied using seven gross phenotypes: survival to pupation, survival to eclosion, development time to pupation and eclosion, thorax length, fecundity, and longevity. We observed interspecific variation among four species, with D. falleni being the most tolerant, followed by D. recens, D. neotestacea, and D. tripunctata, in that order. The results also revealed geographical variation and intraspecific genetic variation in mycotoxin tolerance. This report provides the foundation for further delineating the genetic mechanisms of the mycotoxin tolerance trait.     

Phylogenetic relationships and macro-evolutionary patterns within the <Emphasis Type="Italic">Drosophila tripunctata</Emphasis> “radiation” (Diptera: Drosophilidae)

Despite previous efforts, the evolutionary history of the immigrans-tripunctata clade remains obscure in part due to its hypothesized origin through a rapid radiation. We performed a supermatrix analysis (3,243 base pairs) coupled with richness patterns, environmental phylogenetic signal and radiation tests in order to address phylogenetic relationships and macro-evolutionary hypotheses within this complex group of species. We propose a well-supported evolutionary scenario for the immigrans-tripunctata clade species, in which the tripunctata “radiation” was monophyletic and subdivided into three main lineages: the first including D. pallidipennis (pallidipennis group) imbedded among members of the tripunctata group; the second clustering the cardini and guarani groups; and the third grouping representatives from the tripunctata, calloptera and guaramunu groups. Therefore, we hypothesize that the tripunctata group encompasses a diphyletic taxon, with one clade including the pallidipennis group and the other showing a close affinity to the calloptera and guaramunu groups. Our results also suggest that niche evolution seems to have played a central role in the evolutionary history of the tripunctata species “radiation” allowing effective dispersion and diversification in the Neotropics, possibly in a southwards direction. Although the data as a whole support the notion that this occurred through rapid and successive speciation events, the radiation hypothesis remains to be further corroborated.     

In Vitro and In Vivo Inhibition of Helicobacter pylori by Lactobacillus casei Strain Shirota

We studied the potential inhibitory effect of Lactobacillus casei strain Shirota (from the fermented milk product Yakult [Yakult Ltd., Tokyo, Japan]) on Helicobacter pylori by using (i) in vitro inhibition assays with H. pylori SS1 (Sydney strain 1) and nine H. pylori clinical isolates and (ii) the in vivo H. pylori SS1 mouse model of infection over a period of 9 months. In vitro activity against H. pylori SS1 and all of the clinical isolates was observed in the presence of viable L. casei strain Shirota cells but not in the cell-free culture supernatant, although there was profound inhibition of urease activity. In vivo experiments were performed by oral administration of L. casei strain Shirota in the water supply over a period of 9 months to 6-week-old C57BL/6 mice previously infected with H. pylori SS1 (study group; n = 25). Appropriate control groups of H. pylori-infected but untreated animals (n = 25) and uninfected animals given L. casei strain Shirota (n = 25) also were included in the study. H. pylori colonization and development of gastritis were assessed at 1, 2, 3, 6, and 9 months postinfection. A significant reduction in the levels of H. pylori colonization was observed in the antrum and body mucosa in vivo in the lactobacillus-treated study group, as assessed by viable cultures, compared to the levels in the H. pylori-infected control group. This reduction was accompanied by a significant decline in the associated chronic and active gastric mucosal inflammation observed at each time point throughout the observation period. A trend toward a decrease in the anti-H. pylori immunoglobulin G response was measured in the serum of the animals treated with lactobacillus, although this decrease was not significant.     

Introduction of soybean to North America by Samuel Bowen in 1765

This paper documents the history of the introduction of the soybean (Glycine max), a domesticate of China, to North America. Henry Yonge, the Surveyor-General of Georgia, planted soybeans on his farm at the request of Samuel Bowen in 1765. Mr. Bowen, a former seaman employed by the East India Company, brought soybeans to Savannah, Georgia, from China via London. From 1766, Mr. Bowen planted soybeans on his plantation “Greenwich” located at Thunderbolt, Georgia. The soybeans were used to manufacture soy sauce and vermicelli (soybean noodles). In addition, he manufactured a sago powder substitute made from sweet potatoes. The 3 products were then exported to England. Samuel Bowen received a patent for his manufacturing inventions for producing these products. Another early introduction of soybeans to North America was by Benjamin Franklin. In 1770 he sent seeds from London to John Bartram in Philadelphia.     

Biodegradation of wood exposed in the marine environment: Evaluation of the hazard posed by marine wood-borers in fifteen European sites

The activity of marine wood-borers causes great destruction in maritime wooden structures. Therefore, the aim of this study was to evaluate the hazard posed by marine wood-borers in fifteen European sites, to assist authorities and researchers concerned with the protection of wood in the sea. In northern Europe, Teredo navalis is the species that poses the highest borer hazard while in the Atlantic coast of southern Europe Lyrodus pedicellatus is the most destructive species, with the exception of two sites in Portugal. In these sites, Limnoria tripunctata was more destructive than L. pedicellatus. In the Mediterranean both T. navalis and L. pedicellatus pose a very high borer hazard to wooden structures.Salinity and temperature emerged as the environmental conditions that best explain the occurrence and abundance of wood boring species in the sites surveyed. Three of the species highlighted in this study are warm water species. Therefore their activity might increase in the future, due to global warming. Considering that wood is still a very valuable material for construction, its use for maritime construction should be favoured. Thus research to improve the durability of wooden materials in the marine environment is of paramount importance.     

青杨脊虎天牛对植物源挥发物的EAG和行为反应

测定了青杨脊虎天牛Xylotrechus rusticus （L.）雌、雄成虫对其寄主杨树中的水杨醛（0.95 μmol/μL）和非寄主植物中0.3 μmol/μL的叶绿醇、0.4 μmol/μL的水芹烯和0.6 μmol/μL的 R 型α-蒎烯、S 型α-蒎烯、S型β-蒎烯、3-蒈烯、罗勒烯、香草烯和松节油等10种植物挥发性气味物质的触角电位（EAG）反应。结果表明,与对照相比,这10种植物挥发物多能引起成虫明显的EAG反应( P<0.05,P<0.01 ),其中雌虫对松节油、水杨醛、R 型α-蒎烯和 S 型α-蒎烯的EAG反应较强; 雄虫对 R 型α-蒎烯的EAG反应最强,松节油次之。根据雌虫对这10种挥发物EAG反应的强弱,进一步测定了雌虫对0.00006、0.0006、0.006、0.06、0.6、0.12 μmol/μL的松节油、R 型α-蒎烯、S 型α-蒎烯以及0.000095、0.00095、0.0095、0.095、0.95、0.19 μmol/μL的水杨醛的EAG和行为反应。结果表明,雌虫对松节油、水杨醛和 R 型α-蒎烯的EAG反应随气味物质浓度的增加而增加,水杨醛浓度增加到0.95 μmol/μL、松节油和 R 型α-蒎烯浓度增加到0.6 μmol/μL以后,EAG反应值趋于平稳;对 S 型α-蒎烯的反应随浓度的增加而呈线性增加。水杨醛浓度低于0.095时,对雌虫没有明显的定向作用( P>0.05 ),高于此浓度时表现为驱避作用( P<0.05 ); 松节油在浓度低于或等于0.6 μmol/μL时对雌虫表现为驱避作用,浓度为0.6时驱避效果最佳（ P<0.01 ）。雌虫对 R 型α-蒎烯和 S 型α-蒎烯没有明显的定向行为反应。     

Host Gene Expression Profiling and In Vivo Cytokine Studies to Characterize the Role of Linezolid and Vancomycin in Methicillin-Resistant Staphylococcus aureus (MRSA) Murine Sepsis Model

Linezolid (L), a potent antibiotic for Methicillin Resistant Staphylococcus aureus (MRSA), inhibits bacterial protein synthesis. By contrast, vancomycin (V) is a cell wall active agent. Here, we used a murine sepsis model to test the hypothesis that L treatment is associated with differences in bacterial and host characteristics as compared to V. Mice were injected with S. aureus USA300, and then intravenously treated with 25 mg/kg of either L or V at 2 hours post infection (hpi). In vivo alpha-hemolysin production was reduced in both L and V-treated mice compared to untreated mice but the reduction did not reach the statistical significance [P = 0.12 for L; P = 0.70 for V). PVL was significantly reduced in L-treated mice compared to untreated mice (P = 0.02). However the reduction of in vivo PVL did not reach the statistical significance in V- treated mice compared to untreated mice (P = 0.27). Both antibiotics significantly reduced IL-1β production [P = 0.001 for L; P = 0.006 for V]. IL-6 was significantly reduced with L but not V antibiotic treatment [P<0.001 for L; P = 0.11 for V]. Neither treatment significantly reduced production of TNF-α. Whole-blood gene expression profiling showed no significant effect of L and V on uninfected mice. In S. aureus-infected mice, L altered the expression of a greater number of genes than V (95 vs. 42; P = 0.001). Pathway analysis for the differentially expressed genes identified toll-like receptor signaling pathway to be common to each S. aureus-infected comparison. Expression of immunomodulatory genes like Cxcl9, Cxcl10, Il1r2, Cd14 and Nfkbia was different among the treatment groups. Glycerolipid metabolism pathway was uniquely associated with L treatment in S. aureus infection. This study demonstrates that, as compared to V, treatment with L is associated with reduced levels of toxin production, differences in host inflammatory response, and distinct host gene expression characteristics in MRSA sepsis.     

Production of raw cassava starch-digestive glucoamylase by Rhizopus sp. in liquid culture

Among about 200 Rhizopus strains isolated in Thailand, Rhizopus sp. MB46 was selected as a producer of raw cassava starch-digestive glucoamylase. Rice bran was effective for the enzyme production in a solid culture as well as wheat bran. Addition of turpentine oil into the rice bran solid culture increased the productivity. Rhizopus sp. MB46 was found to produce glucoamylase in a liquid culture containing 1% rice bran but not in one consisting of 10% raw cassava starch of 2% glucose. The productivity per 1 g solids in the medium in liquid culture was finally improved 6-times by utilization of n-hexane-treated rice bran, supplement of 0.1% meat extract and addition of gauze as a support. The activity was superior to that in turpentine oil-supplemented solid culture.     

Evidence for a high molecular weight pre-robustoxin molecule in the venom of the male Sydney funnel-web spider (Atrax robustus)

Robustoxin is the lethal polypeptide toxin in Atrax robustus venom. A monoclonal antibody was produced using synthetic, unfolded robustoxin conjugated to keyhole limpet haemocyanin as the immunogen. This monoclonal antibody did not protect newborn mice against challenge with the crude venom of the male Sydney funnel-web spider, but did slightly prolong their survival time. Western blotted crude venom of the male Sydney funnel-web spider showed two monoclonal antibody binding bands. One band at low M_r corresponded to robustoxin (M_r 4854), while the other higher M_r band (approximately 37,000) may be due to a pre-robustoxin molecule.     

Effects in s.e. Queensland during 1967–72 of insects introduced to controlLantana camara

Seasonal fluctuations in populations of the introduced speciesTeleonemia scrupulosa Stål and damage by this insect to common taxa ofLantana camara, were observed at several field sites and experimental plots. At 2 plots effects ofT. scrupulosa populations on untreated plants were compared with plants protected by application of insecticides. On untreated plants damage varied seasonally and between taxa ofL. camara. In general attack byT. scrupulosa reduced fruiting and caused severe short term foliage damage. Combined with severe stress from some other cause heavy attack may result in long term damage or death to plants. However Common PinkL. camara was more resistant to damage than were the other taxa studied. At the time of these experiments other insect species were at very low population levels and did not have observable effects onL. camara.     

Community deployment of a synthetic pheromone of the sand fly Lutzomyia longipalpis co-located with insecticide reduces vector abundance in treated and neighbouring untreated houses: Implications for control of Leishmania infantum

BackgroundThe rising incidence of visceral leishmaniasis due to Leishmania infantum requires novel methods to control transmission by the sand fly vector. Indoor residual spraying of insecticide (IRS) against these largely exophilic / exophagic vectors may not be the most effective method. A synthetic copy of the male sex-aggregation pheromone of the key vector species Lutzomyia longipalpis in the Americas, was co-located with residual pyrethroid insecticide, and tested for its effects on vector abundance, hence potential transmission, in a Brazilian community study.MethodsHouses within eight defined semi-urban blocks in an endemic municipality in Brazil were randomised to synthetic pheromone + insecticide or to placebo treatments. A similar number of houses located >100m from each block were placebo treated and considered as “True Controls” (thus, analysed as three trial arms). Insecticide was sprayed on a 2.6m² surface area of the property boundary or outbuilding wall, co-located within one metre of 50mg synthetic pheromone in controlled-release dispensers. Vector numbers captured in nearby CDC light traps were recorded at monthly intervals over 3 months post intervention. Recruited sentinel houses under True Control and pheromone + insecticide treatments were similarly monitored at 7–9 day intervals. The intervention effects were estimated by mixed effects negative binomial models compared to the True Control group.ResultsDose-response field assays using 50mg of the synthetic pheromone captured a mean 4.8 (95% C.L.: 3.91, 5.80) to 6.3 (95% C.L.: 3.24, 12.11) times more vectors (female Lu. longipalpis) than using 10mg of synthetic pheromone. The intervention reduced household female vector abundance by 59% (C.L.: 48.7, 66.7%) (IRR = 0.41) estimated by the cross-sectional community study, and by 70% (C.L.: 56.7%, 78.8%) estimated by the longitudinal sentinel study. Similar reductions in male Lu. longipalpis were observed. Beneficial spill-over intervention effects were also observed at nearby untreated households with a mean reduction of 24% (95% C.L.: 0.050%, 39.8%) in female vectors. The spill-over effect in untreated houses was 44% (95% C.L.: 29.7%, 56.1%) as effective as the intervention in pheromone-treated houses. Ownership of chickens increased the intervention effects in both treated and untreated houses, attributed to the suspected synergistic attraction of the synthetic pheromone and chicken kairomones. The variation in IRR between study blocks was not associated with inter-household distances, household densities, or coverage (proportion of total households treated).ConclusionsThe study confirms the entomological efficacy of the lure-and-kill method to reduce the abundance of this important sand fly vector in treated and untreated homesteads. The outcomes were achieved by low coverage and using only 1–2% of the quantity of insecticide as normally required for IRS, indicating the potential cost-effectiveness of this method. Implications for programmatic deployment of this vector control method are discussed.     

The acute phase response of cod (Gadus morhua L.): expression of immune response genes

An acute phase response (APR) was experimentally induced in Atlantic cod (Gadus morhua L.) by intramuscular injection of turpentine oil. The change in the expression of immune related genes was monitored in the anterior kidney and the spleen over a period of 7 days. The genes examined were two types of pentraxins, apolipoprotein A1 (ApoA-I), the complement component C3, interleukin-1β (IL-1β), transferrin, cathelicidin, and hepcidin. All genes were constitutively expressed in both organs and their expression amplified by the turpentine injection. A pattern of response was observed both with respect to the organ preference and to the timing of a maximum response. The increased gene expression of the pentraxins, ApoA-I and C3 was restricted to the anterior kidney, the gene expression of IL-1β, cathelicidin, and transferrin increased in both organs, while hepcidin gene expression was only significantly increased in the spleen. The pentraxins and ApoA-I appear to be early mediators of APR in cod, possibly stimulating C3 and IL-1β response, while the antimicrobial peptides may play a minor role. The increase in transferrin gene expression in both organs, and apparent indifference to cortisol release associated with the turpentine injection, suggests that this could be a typical acute phase protein in cod.