Leishmania amazonensis: effects of oral treatment with copaiba oil in mice

Leishmaniasis is a severe public-health problem, with high rates of morbidity and mortality. Efforts to find new, effective and safe oral agents for the treatment of leishmaniasis have been ongoing for several decades, in order to avoid the problems with the currently used antimonials. In the present study, we found that a copaiba oil oral treatment (Group IV) caused a significant reduction in the average lesion size (1.1 ± 0.4 mm) against Leishmania amazonensis lesions compared with untreated mice (Group I) (4.4 ± 1.3 mm). To prove the safety of the oil, the toxicity and genotoxicity were also determined. Histopathological evaluation did not reveal changes in the copaiba oil-treated animals compared to the control animals. In the mutagenicity evaluation, (micronucleus test) the dose tested (2000 mg/kg) showed no genotoxic effects. Morphological and ultrastructural analyses demonstrated notable changes in parasite cells treated with this oleoresin. The main ultrastructural effect was mitochondrial swelling. We also demonstrated that in vitro copaiba oil treatment of L. amazonensis led to an increase in plasma membrane permeability, and depolarization in the mitochondrial membrane potential in parasite cells. Although the mechanism of action of the oleoresin is still unclear, these findings indicate that copaiba oil is a possible new drug, which would provide a safer, shorter, less-expensive, and more easily administered treatment for leishmaniasis.     

The reproductive system of Hippolyte niezabitowskii (Decapoda, Caridea)

Data on the reproductive biology of the genus Hippolyte are available from studies of Hippolyte inermis. These findings support the idea that these species are protandric hermaphrodites, exhibiting sex reversal. However, recent studies of this and other species, such as Hippolyte obliquimanus and Hippolyte williamsi, have not reported sex reversal and suggest a gonochoric condition. In the present study, histological analyses were conducted of the ovaries, testes, oviducts, vasa deferentia and the development of the male appendage. The results of this study show that the Hippolyte niezabitowskii population from southern Spain should also be viewed as a gonochoric species. The study found no evidence of sex reversals. These conclusions are also supported by information on the structure of the population and by the results of studies of the male appendage. The population structure involves different size distributions of males and of females. Size classes 2-14 exhibit substantial overlap. No evidence of sex reversal was obtained from the study of the male appendage.     

The reproductive performance of female Formosan sambar deer (Cervus unicolor swinhoei) in semi-domesticated herds

This study documented the reproductive performance of 210 adult female Formosan sambar deer (FSD, Cervus unicolor swinhoei) from four semi-domesticated deer herds in Taiwan. An extensive analysis of 525 reproductive records from 2000 to 2008, including the conditions of estrus, gestation, and parturition was conducted. The mean ± S.E.M. lengths of the estrous cycle, gestation, and fawning interval were 18.2 ± 0.5 d (n = 56), 258.6 ± 0.3 d (n = 160), and 369.9 ± 2.3 d (n = 122), respectively. Hand breeding was performed between June and December (n = 494), with the majority (93.1%) occurring between July and October (P < 0.05). Fawning occurred from February to September (n = 318), and most frequently (83.0%) between April and June (P < 0.05). Pregnancy rate per mating in FSD hinds was 64.4%. There was a 1.3:1 male-to-female ratio at birth (P < 0.05) among 320 fawns, and only two cases of twinning (0.63%). The postnatal mortality rate was 6.6% (21/320), and the mortality rate in fawns before weaning did not exceed 8% on any farm. Fecundity was enhanced by high pregnancy rates and high offspring survival rates. This study provides baseline information on reproductive performance of FSD, which should be valuable to veterinarians and deer industry personnel for management of FSD on farms in subtropical countries.     

Notes on the reproductive biology of Phrynus exsul Harvey, 2002 (Arachnida: Amblypygi: Phrynidae)

Mating behaviour and spermatophore morphology of the only Old World species of the Phrynidae, Phrynus exsul Harvey, 2002, are described and compared to the reproductive biology of other whip spiders. Mating behaviour and spermatophore morphology clearly show that P. exsul is correctly included in the Phrynidae. But some details and other characters suggest that P.exsul has no close relation with any of the Neotropical species of Phrynus Lamarck, 1793, probably having been evolutionary distinct for a long time from the Neotropical species of Phrynus. Its occurrence on the Indonesian Island Flores thus remains an enigma; is P. exsul a relic from a more global distribution of the genus Phrynus in the past or the result of prehistoric rafting across the ocean?     

Presence and titer of methyl palmitate in the Medfly (Ceratitis capitata) during reproductive maturation

The relative amounts of methyl palmitate (MP) during the first 10 days post-eclosion were determined in whole-body extracts of adult female Ceratitis capitata by SIM monitoring of the 74 m/z fragment. MP peaks in receptive 3-day-old virgin females coincide with previously reported production of Juvenile Hormone (JH) by the corpus allatum (CA). Mating in the Medfly induces female non-receptivity. Indirect evidence suggests that the mevalonate pathway to sesquiterpene biosynthesis is underdeveloped in newly eclosed females. We propose that the pathway leading to synthesis of JH is markedly diverted in non-receptive virgin females to fatty acid synthesis, and partly so-in non-receptive mated females, leading to production of palmitic acid, presumably methylated thereafter. MP is depressed and remains marginal thereafter for the 7 days examined in the virgin female but goes through an apparent second cycle in the mated female. This contrasts with the consistent increase of allatal biosynthesis of MP of virgin and mated females previously reported and suggests additional control mechanisms in vivo. During the period of reduced receptivity following the first mating a second apparent peak of MP is observed. MP is a metabolic default metabolite of reproductively immature females whose putative role in reproductive physiology remains to be defined.     

Structural alterations in the male reproductive system of the freshwater crayfish, Cherax quadricarinatus (Decapoda, Parastacidae)

No diseases affecting reproductive performance have been previously reported in freshwater crayfishes. This study aims to characterise one reproductive system abnormality found in males of Cherax quadricarinatus reared in captivity. Fifteen adult males of C. quadricarinatus (70-110 g) were purchased from San Mateo S.A. farm (Entre Ríos, Argentina) each season during 2007. Macroscopic analysis showed that 26.6% of the animals sacrificed in winter presented brownish distal vasa deferentia. Histological analysis showed different levels of structural abnormality in the epithelium of the vasa deferentia and spermatophore. Granular and hyaline haemocytes were identified within the vasa deferentia but no significant differences were found in the sperm count between normal and brownish vas deferens. Histological analysis of the crayfishes sacrificed in autumn also showed these modifications in 22% of the animals, however, they did not show the brownish colour under macroscopic analysis. The similarities between the male reproductive system syndrome in shrimps and the abnormalities found in C. quadricarinatus are notable. An unspecific response to thermic stress is a possible explanation of these structural alterations.     

Temporal variability of abundance and reproductive traits of Centropages kroyeri (Calanoida; Copepoda) in Bizerte Channel (SW Mediterranean Sea, Tunisia)

Abundance and reproductive traits (spawning female, egg production, egg hatching success) and naupliar survival of the dominant calanoid copepod Centropages kroyeri were investigated in relation to biotic (chlorophyll a, phytoplankton and microplankton) and abiotic (temperature and salinity) parameters at a monitoring station located in Bizerte Channel, northern Tunisia. The monitoring was carried out between July 2004 and December 2004, according to two temporal sampling strategies. First, during July, the supposed maximum abundance period of C. kroyeri, daily sampling was conducted. Thereafter, sampling was carried out weekly until the end of the sampling period.The temporal variability of the environmental factors in relation to the reproductive traits of C. kroyeri revealed, first, that temperature is important in controlling the reproduction and dynamics of this species, and second, that the quantity and quality of food available in the field also strongly influence its reproduction. On several sampling dates, the output of certain reproductive traits such as hatching success after 24 h was very weak. It is possible that C. kroyeri produced resting eggs during the study period. Nerveless, the analysis of the phytoplankton during these periods showed that low reproductive activity may have been caused by low phytoplankton quantity and/or by the dominant presence of certain diatom species that are supposed to have an inhibitory effect on the reproductive traits of C. kroyeri, especially on egg hatching success.Among the considered external parameters, temperature and food quality and availability seem to be the most important factors for the reproduction and dynamics of C. kroyeri. Under these biotic and abiotic field conditions, this copepod seems able to modulate the abundance of its population while controlling its reproduction.     

Selective attraction and reproductive performance of a harpacticoid copepod in a response to biofilms

The relationship between monobacterial films and the preference of harpacticoid copepods for such films was investigated using still water multiple-choice assays with natural biofilm and sterile conditions as controls. Adult Schizopera sp. were most attracted by a heterogeneous natural biofilm, followed by monospecies-biofilms of Rhodovulum sp., Vibrio proteolyticus, and Flexibacter sp. The preferred bacterial films stemmed from different phylogenetic and physiological groups. The results indicated that the harpacticoid Schizopera sp. was effectively and differentially attracted by bacterial films. Since bacteria constitute a substantial portion of the organic carbon available at the sea bottom as nutritive sources for harpacticoid copepods, we subsequently examined the influence of 9 bacterial strains and a natural biofilm as a nutrient source on the growth and reproductive performance of ontogenetic stages (nauplii and copepodids) of Schizopera sp. The food value of bacterial strains was assayed in terms of life table data that provided growth parameters. All variables were affected by the type of food offered. A diet on Rhodovulum sp. resulted in optimal growth performance of nauplii and copepodids demonstrating that bacteria can be used as a sole diet to support postembryonic development. The present study is the first to link behavioral preferences to bacterial biofilms with life history parameters when cultivating harpacticoid copepods on the same bacterial strains as the only diet. This study revealed a discrepancy between the biofilm favored (natural biofilm) and the one leading to maximal reproductive performance (monobacterial film of Rhodovulum sp. MB253) as indicated by major life table data as net reproductive rate (R_o), mean generation time (T_m), and capacity for increase (r_c).     

The oleoresin secretory system in seedlings and adult plants of copaíba (Copaifera langsdorffii Desf., Leguminosae-Caesalpinioideae)

The ecological and economic importance of oleoresin produced by Copaifera langsdorffii is well established. This study aims to investigate the ontogeny, anatomy and ultrastructure of the internal glands of C. langsdorffii during plant development. Samples were processed for light and electron microscopy and a specific technique was applied to impregnate endomembranes. Internal secretory glands were observed in the hypocotyl, epicotyl and eophylls of seedlings, and in the primary stem, pulvinus, petiole, rachis and leaf blade of adult plants. Canals and cavities show differential distribution. They arise from ground meristem cells, and the lumen is first formed by schizogenesis followed by later schizolysigenous development. The dense cytoplasm of epithelial cells shows mitochondria, plastids without thylakoids, polyribosomes and endoplasmic reticulum. A periplastidial reticulum was also observed. Secretion is released by eccrine, granulocrine and holocrine processes. Lipophilic and hydrophilic compounds were histochemically detected in both canals and cavities, whereas resin was detected only in canals. The presence of these substances has been associated with plants’ defences against dehydration, as well as against attacks from herbivores and pathogens, from seedling stage onwards.     

The reproductive biology of the genusBaldellia (Alismataceae)

The reproductive biology of the genusBaldellia was studied under cultivation and in nature.B. ranunculoides subsp.repens is self-incompatible whereasB. ranunculoides subsp.ranunculoides andB. alpestris are self-compatible. Bud-pollination occurs in theB. alpestris. Self-incompatibility correlates with the production of fewer, larger, more showy flowers, higher pollen production (but reduced pollen grain size), fewer carpels and smaller seeds. Resource allocation to sexual reproduction is inversely correlated with the degree of vegetative growth.B. ranunculoides subsp.repens produces more ramets than subsp.ranunculoides, which reproduces mainly by seeds.B. alpestris is intermediate between the two since it is self-compatible, produces fewer, larger seed than subsp.ranunculoides and is also able to propagate by vegetative means.     

Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control

Lavandula angustifolia is a well known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of L. angustifolia essential oil (EO) against engorged Rhipicephalus (Boophilus) annulatus (Acari; Ixodidae) females. For this purpose six serial concentrations (0.5, 1.0, 2.0, 4.0, 6.0 and 8.0% w/v) of L. angustifolia EO were used. There was considerable mortality in concentrations more than 4.0% although there were no differences between 6.0 and 8.0% in all measured criteria. The mortality rate 24 h after inoculation was 73.26 and 100% in groups treated with 4.0 and 8.0% EO, respectively. Lavender EO also reduced tick egg weight in a concentration-dependent manner. The amount of eggs produced varied from 0.12 g (at 0.5% EO) to 0.00 g (at 8.0% EO) but did not differ statistically from the control. L. angustifolia EO caused 100% failure in egg laying at 6.0 and 8.0% whereas this value in the control group was zero. A positive correlation between L. angustifolia EO concentration and tick control, assessed by relative mortality rate and egg-laying weight, was observed by the EO LC/EC₅₀, which, when calculated using the Probit test, was 2.76-fold higher than the control. Lavender is a promising acaricidal against R. (B.) annulatusin vitro.     

Virulence, horizontal transmission, and sublethal reproductive effects of Metarhizium anisopliae (Anamorphic fungi) on the German cockroach (Blattodea: Blattellidae)

Virulence of Metarhizium anisopliae (Metschnikoff) Sorokin strain EAMa 01/121-Su against the German Cockroach, Blatella germanica (L.), was determined using four concentrations ranging from 4.2 x 10(6) to 4.2 x 10(9) spores per milliliter. The LD50 value was 1.4 x 10(7) spores per milliliter (56,000 spores per cockroach) and LT50 values were 14.8 days and 5.3 days for 4.2 x 10(8) and 4.2 x 10(9) spores per milliliter, respectively. An experiment was conducted to evaluate whether a fungal transmission could exist among infected and healthy cockroaches. Percentage mortality at a ratio of 1:10 of infected to unexposed cockroaches was 87.5% and LT50 was 12.2 days, which indicated the potential of this strain to be horizontally transmitted and to rapidly spread the infection in the insect population. The effect of a sublethal dose (ca. LD60) of M. anisopliae EAMa 01/121-Su strain, applied topically on German cockroaches, was studied by reciprocal crossing. Othecal production, oothecal hatchability, and nymphal production declined upon exposure to M. anisopliae EAMa 01/121-Su strain. The mean number of oothecae laid by female was progressively and significantly reduced by fungal treatment from second oviposition period onwards. Oothecal hatch of fungally challenged females was reduced by 46-49%, oothecal viability by 48-85%, and nymphal production by 22-35%. Only treated females showed an effect on oothecal production, oothecal hatch, and nymphal production, although oothecal hatch was also governed by treated males at a higher significance level. Our results on virulence and horizontal transmission of fungal conidia of M. anisopliae EAMa 01/121-Su strain and its sublethal reproductive effects on German cockroach females are discussed in terms of its potential to decrease the pest status of B. germanica in the short and long terms.     

Rh(I) complexes containing tris(pyrazolyl)amine and bis(pyrazolyl)amine ligands: synthesis and NMR studies

The tris(pyrazolyl)amine ligands: tris[2-(1-pyrazolyl)methyl]amine (tpma), tris [3,5-dimethyl-1-pyrazolyl)methyl]amine (tdma), tris[2-(1-pyrazolyl)ethyl]amine (tpea), tris[2-(3,5-dimethyl-1-pyrazolyl)ethyl]amine (tdea) and bis(pyrazolyl)amine ligands: bis[2-(1-pyrazolyl)ethyl]amine (bpea) and bis[2-(3,5-dimethyl-1-pyrazolyl)ethyl]amine (bdea) react with [RhCl(cod)]₂ in presence of NaBF₄ (tpma, tdma and bdea) or AgBF₄ (tpea, tdea and bpea) to lead to [Rh(cod)L] (BF₄) (L=tpma (1), tdma (2), bdea (3), tpea (4), tdea (5) and bpea (6)). These complexes have been characterised by elemental analyses, conductivity, IR, ¹H and ¹³C NMR spectroscopy and liquid mass (with electrospray) spectrometry. The ¹H NMR spectra of 1, 2 show the presence of two isomers in solution in a 3:1 ratio (coordination κ² or κ³ type) in a thermodynamic equilibrium. The steric bulk of cyclo-octa-1,5-diene causes it to prefer the κ² mode of bonding as majority. Similar to previous published results, complexes 4 and 5 exist in a sole form in solution (probably κ² isomer). Finally, the complexes 3 and 6 are fluxional. A NMR study shows that this fluxional process is not frozen at 183 K.     

Controlled trial of induction of labor by vaginal suppositories containing prostaglandin E2

A group of 84 women at 39 – 43 weeks of pregnancy were randomly allocated to a blind trial of induction of labor with vaginal suppositories containing inert material or either 0.2 mg or 0.4 mg of prostaglandin E₂. The suppositories were self-administered every two hours during waking hours on two successive days until labor started or 15 had been used. Side-effects were absent. Labor was established within 48 hr of insertion of the first suppository in 9.3% of control patients, 65.4% of those treated with 0.2 mg PGE₂ and 85.7% of those treated with 0.4 mg PGE₂. The mean Apgar scores in the three groups were the same. The mean total dose of PGE₂ were 2.0 mg (0.2 mg group) and 2.3 mg (0.4 mg group). It is concluded that vaginal PGE₂ is an effective and acceptable method of inducing labor at term.     

Food utilization efficiency in fifth instar larvae of Antheraea mylitta (Lepidoptera: Saturniidae) infected with Nosema sp. and its effect on reproductive potential and silk production

Antheraea mylitta, a sericigenous insect of economical importance is often infected with an intracellular parasite of the genus Nosema. This pathogen is known to cause fatal pebrine disease and is considered as an important factor that strongly influences the development of the host. Larvae developed from the eggs laid by a female infected with Nosema sp. showed extended development period. The increment in the larval weight declined significantly in infected larvae in comparison to uninfected ones. Food consumption, digestion, relative consumption rate (RCR), efficiency of conversion of ingested food (ECI), efficiency of conversion of digested food (ECD), and relative growth rate (RGR) values declined significantly, but at the same time a significant increase in approximate digestibility (AD) was also observed. Silk production declined in infected larvae. Silk gland weight and shell weight also significantly declined following infection over uninfected larvae. The reproductive potential in adults declined significantly (P<0.001) with decrease in ovary weight (31.6%), fecundity (54.1%), and fertility (34.9%). Egg chorionation was also affected in adults, which developed from infected larvae. The maternal infection level in one generation (10.4 x 10(6) spores/female) decreased significantly in the next generation (8.0 x 10(6) spores/female).     

Insecticidal effects of Flourensia oolepis Blake (Asteraceae) essential oil

Flourensia oolepis Blake (Asteraceae) essential oil had a complex chemical composition with τ-muurolene (6.14%), santolinetriene (6.22%), 2-methylene-4,8,8-trimethyl-4-vinyl-bicyclo[5.2.0]nonane (10.15%), δ-cadinene (10.27%) and γ-gurjunene (20.69%) comprising more than 50% of the oil. This oil had repellent and toxic effects on Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) adults, acting as a contact toxin. Myzus persicae (Sulzer) (Homoptera: Aphididae) and Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae) adults showed behavioral sensibility to this oil.     

Candida albicans strain-dependent modulation of pro-inflammatory cytokine release by in vitro oral and vaginal mucosal models

Candida albicans is a commensal organism at several sites and is a versatile, opportunistic pathogen. The underlying factors of pathogen and host associated with commensalism and pathogenicity in C. albicans are complex and their importance is largely unknown. We aimed to study the responses of oral epithelial (OEM) and vaginal epithelial models (VEM) to infection by oral and vaginal C. albicans strains to obtain evidence of inter-strain differences in pathogenicity and of site-specificity. Following inoculation of models, proinflammatory cytokines IL-1α, IL-1β, IL-6, IL-8 and prostaglandin E2 (PGE2) release were monitored and histological staining undertaken. Striking differences in strain behaviour and epithelial responses were observed. IL-1α, IL-1β and IL-8 release were significantly increased from the OEM in response to denture stomatitis strain NCYC 1467. Increased IL-8 release also followed infection of the OEM with both vaginal strains. Overall the VEM was relatively unresponsive to infection with either oral or vaginal strains under these conditions. Adherence and hyphal development were observed for all strains on both models although extensive, uniform tissue penetration was seen only with stomatitis strain NCYC 1467 on the OEM. Candidal strains were assayed for phospholipase (PL) and secreted aspartyl proteinase (SAP) activities where phospholipase (PL) activity was highest for strain NCYC 1467 although highest SAP activity was observed for vaginal strain NCPF 8112 in this assay. This is the first study to concurrently investigate cytokine production from oral and epithelial models using candidal strains originating from these respective mucosal sites from healthy and disease states. These data demonstrate significant differences in inflammatory responses of host epithelia to individual C. albicans strains.     

Number of reproductive cycles of Varroa jacobsoni in honey-bee (Apis mellifera) colonies

Very little data exists concerning the number of reproductive cycles performed by individual Varroa mites. To understand the population dynamics of the Varroa mite it is necessary to know the number of fertile female offspring each Varroa female produces during her lifetime. The lifetime reproduction capacity of the mite consists of the mean number of fertile female offspring produced during each reproductive cycle multiplied by the mean number of cell passages. This paper describes an experimental design to estimate the number of reproductive cycles where mites are transferred to new mite-free colonies for reproduction in sealed brood cells. The data presented suggests that the mean number of reproductive cycles performed by the individual female mite is larger than previously accepted. Under optimal conditions, the mean number of reproductive cycles by Varroa females is probably greater than 1.5 but less than 2. Furthermore, the results show that the reproductive success of Varroa females going into cells to reproduce is not influenced by previous brood cycles.