Effectiveness of hyperbaric oxygen for experimental treatment of schistosomiasis mansoni using praziquantel-free and encapsulated into liposomes: Assay in adult worms and oviposition

The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

Serum protein profile of hookworm infection in dogs

Hookworms are parasitic nematodes that cause anemia and intestinal infections in dogs, especially with large worm burdens. However, the serum protein profile of this parasitological disease is still poorly understood. The present study was design to evaluate 80 asymptomatic dogs (age; 8 months–2 years) to detect the presence of the hookworm thin-shelled, morulated eggs in faeces using faecal flotation and to evaluate the serum protein fractions determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Forty dogs had an elevated quantity of eggs in faeces (+++) (PD) and 40 dogs were healthy (HD). Infected dogs showed significant increases for IgG estimated concentrations (PD 1.79±0.8 g/dL and HD 1.44±0.72 g/dL, p= 0.04), for ceruloplasmin estimated concentrations (PD 19±15 mg/dL and HD 5±3.5 mg/dL, p=0.0001), for alpha 1-acid glycoprotein estimated concentrations (PD 31.4±17.9 mg/dL and HD 13.5±12.1 mg/dL, p=0.0001) and for a non-identified protein of 23 kDa estimated concentrations (PD 641.5±194.9 mg/dL and HD 519.8± 197.9 mg/dL, p=0.007). Dogs with hookworm infection showed significant differences in the serum protein profile when compared to healthy animals.

Studies on the Helminth Fauna of Alaska. XLI. Observations on Cestodes of the Genus Diplogonoporus Lönnberg, 1892 (Diphyllobothriidae)

The study of a collection of cestodes assigned to the genus Diplogonoporus Lönnberg, 1892 disclosed but two species, D. balaenopterae Lönnberg, 1892, and D. tetrapterus (von Siebold, 1848) (provis.). These cestodes occur characteristically in marine mammals but occasionally are found in terrestrial hosts; D. balaenopterae is recorded for the first time from the domestic dog, and it is concluded that D. grandis (Blanchard, 1894), from man, is conspecific with D. balaenopterae. The latter species is recorded for the first time from the humpback whale, Megaptera novaeangliae (Borowski). The relatively small D. tetrapterus, a common parasite of the Steller sea lion, Eumetopias jubata (Schreber), is reported for the first time from the sea otter, Enhydra lutris Linnaeus, and from the domestic mink, Mustela vison Schreber. Descriptions of representative specimens are presented, and the taxonomic status of other species assigned to Diplogonoporus is discussed. Although the diplogonadic organization of these cestodes is somewhat variable, it is nevertheless constant and serves to characterize the genus Diplogonoporus. The process of asexual reproduction by means of transverse subdivision of primary segments is described. This ability and the diplogonadic structure of these cestodes are considered to be adaptations that increase the production of eggs and thereby the probability of reproductive success in the marine habitat.

Development of Nicarbazin as a Reproductive Inhibitor for Resident Canada Geese

Expanding populations of resident Canada geese that remain in suburban and urban areas year-round often result in increased conflicts with humans. Non-lethal and humane means are needed for managing the size of Canada goose flocks residing near or on airports, golf courses, industrial parks, government sites, and city parks. A side effect of nicarbazin, a veterinary drug used to control coccidiosis in chickens, is decreased egg production and hatching. Exploiting this side effect, studies of nicarbazin for reducing the hatchability of eggs from Canada geese were conducted. An initial study in Coturnix quail verified reduction in hatchability in a species other than chickens. Because plasma nicarbazin was not routinely measured, a study in chickens was conducted to determine the relationship between plasma and egg nicarbazin. A comparative study in chickens, mallards, and Canada geese showed that nicarbazin absorption was lowest in geese. Studies in both penned and wild Canada geese showed that reduction in hatchability was possible but neither study used bait suitable for general field application. Bait development led to the OvoControl-G® (Innolytics LLC) bait, which resulted in reduction in hatchability of 51% at treated sites compared to control sites in the field. Previous studies showed that nicarbazin is practically non-toxic and is environmentally friendly; timing and management of baiting will minimize non-target hazards. OvoControl-G® 2500 ppm nicarbazin bait is recommended for incorporation into a comprehensive management plan as a reproductive inhibitor for use in controlling resident Canada goose flock sizes.

SIGNIFICANCE AND CONTROL OF FUNGAL DISEASES RELATED TO BIRD ROOSTS

Certain fungi have been found frequently as saprophytes in areas containing large amounts of bird excreta. These fungi have the ability to survive, multiply, and cause disease once they have entered a host. Two of these are Crypto-coccus neoformans and Histoplasma capsulatum. Both may easily become airborne and be disseminated throughout an area by the prevailing winds. C. neo-formans is commonly isolated from the excreta of pigeon habitats, and in turn has been associated with clinical cases of cryptococcosis, while blackbird roosts, harboring H. capsulatum, have been responsible for several outbreaks of histoplasmosis. When either of these fungi have become established in nature, the sites may become foci for infection and epidemics may occur if the sites are disturbed. This has led to investigation of these organisms with respect to: 1) the frequency of isolation of H. capsulatum from the soil beneath blackbird roosts in a histoplasmosis endemic area; 2) the infectivity of undisturbed roosts positive for H. capsulatum; and 3) the effectiveness of chemical decontamination of areas containing C. neoformans or H. capsulatum.

Intraspecific competition in Zabrotes subfasciatus: Physiological and behavioral adaptations to different amounts of host

The effects of competition on populations of the bean weevil Zabrotes subfasciatus were analyzed during 41 generations under different competition levels. Three competition environments were established by maintaining the number of couples (6) and varying the amount of available host seeds: HC, high (limited availability of host: 1.35 g); IC, intermediate (intermediate availability of host: 6 g); and LC, low competition (abundance of host: 36 g). It was found that the distribution of the eggs laid on grains was different among treatments: in LC, for example, although females showed high fecundity (35.4 +/- 5.6 eggs/female) the number of eggs laid on each grain was small (1.2 +/- 0.4 eggs on each seed), thus avoiding larval competition of their offspring; whereas in HC treatment, females showed low fecundity (27.04 +/- 4.5 eggs/female) but laid many eggs on each grain (15.03 +/- 4.3 eggs). There were no changes in the ability to respond to different amounts of host via oviposition behavior (egg distribution) during 41 generations. However, HC females had more offspring than LC females under HC conditions. This suggests that HC insects evolved toward higher fitness in crowded conditions. In addition, after inverting the competition level, insects behaved independently of the treatment conditions they experienced through generations, thus showing that oviposition behavior is flexible. Taken together, our results show that Z. subfasciatus presents a broad range of behavioral and physiological responses which allows for quick and reversible adjustments to sudden changes in the amount of resources.

Mating behaviour, nympho-imaginal development and description of a new Mesabolivar species (Araneae: Pholcidae) from the Brazilian dry forest

A new species of Mesabolivar is described from Brazilian forests: Mesabolivar delclaroi. The mating behaviour, postembryonic development, number of egg sacs and eggs, birth rate, number of instars, developmental time until adulthood, sex ratio and size of cephalothorax (per instar) were recorded. The sexual behaviour was described and categorized into four steps: courtship, pre-copulation, copulation and post-copulation. After hatching, individuals presented five instars until maturity. The mean number of eggs (42 +/- 16.6) and live births (31.5 +/- 3.4) of the first egg sac were significantly greater than that of a second one (23.8 +/- 3.8, and 19.25 +/- 3.9, respectively). The developmental time from birth to adulthood (130.8 +/- 9.6 days) did not differ significantly between egg sacs produced (128.61 +/- 11.1). The size of the cephalothorax did not differ among adults or between sexes. The sex ratio revealed a shift in favour of females (4: 3).

Morphology and biological aspects of Gundlachia ticaga from SE Brazil (Gastropoda: Basommatophora: Ancylidae)

Some aspects of the biology, reproduction and morphology of Gundlachia ticaga (MARCUS & MARCUS 1962) were studied and compared with literature data. A sample of similar to 100 specimens, collected in a darn in Ribeirao Preto, were maintained in Petri dishes at the laboratory during a period no longer than a month. The oviposition, number of capsules, and incubation time of eggs were registered for each individual. A morphologic study was carried out in both, alive and fixed specimens. Although some morphological discrepancies were identified, the majority of data confirms what was previously described for the species. Regarding the biology, however, the results were somewhat different from those previously reported. The incubation time of the eggs varied from 10 to 13 days, and the number of embryos in the interior of the ovigerous capsules varied from 1 to 4. In addition, each individual laid 9 eggs during the first month of the reproductive period. The formation of septa occurred in some animals and intermediary phases are figured.

POPULATION DENSITY OF THE HELMETED CURASSOW (PAUXI PAUXI) IN TAMA NATIONAL PARK, COLOMBIA

We estimated the population density of the Helmeted Curassow (Pauxi pauxi) in Tama National Park (TNP) Colombia, using visual counts between December 2006 and December 2008. We used six line transects (1 km each) equitably distributed in a natural forest between 800 and 1,200 m asl in the southern part of the park. The sampling effort was 588 hrs with a total distance of 490 km, a detection rate of 0.06 records/hr, and an encounter rate of 0.08 individuals/km. Only solitary individuals were recorded (n = 40); the estimated density was 4.8 individuals/km(2). Most detections occurred in the lower strata of the forest (floor and sub-canopy) where hunters take advantage of curassows in the lower strata for successful harvest. The southern sector of TNP becomes important in the dry season. Our study suggests a large population is in TNP, but harvesting activities including removal of eggs, chicks, and juveniles, and hunting adults are affecting the reproductive rate and population of the species. Received 6 June 2011. Accepted 2 February 2012.

Vitellogenin content in fat body and ovary homogenates of workers and queens of Melipona quadrifasciata anthidioides during vitellogenesis

Vitellogenin (Vg) is an egg yolk protein that is produced primarily in the fat body of most female insects. In the advanced social structure of eusocial honeybees, the presence of the queen inhibits egg maturation in the workers ovaries. However in the stingless bee Melipona quadrifasciata, the workers always develop ovaries and lay a certain amount of eggs while provisioning the brood cells with larval food during what is known as the worker nurse phase. The present work is a comparative study of the presence of Vg in homogenates of the fat bodies and ovaries of the nurse workers, and the virgin and physogastric queens of M. quadrifasciata. The presence of Vg was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting using Apis mellifera anti-egg antibody. Vg was not detected in the fat bodies or ovaries of the workers, but it was found in the ovaries of virgin and physogastric queens and in the fat body of physogastric queens. The results are discussed, taking into account the reproductive state of the individuals and the other possible roles of Vg, such as a storage protein for metoabolism of other organs.

Embryonic development and larval stages of Steindachneridion parahybae (Siluriformes: Pimelodidae) - implications for the conservation and rearing of this endangered Neotropical species

Steindachneridion parahybae is a freshwater catfish endemic to the Paraiba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the embryonic and larval development of S. parahybae in captivity, with emphasis in major events during the ontogeny of S. parahybae. Broodstocks were artificially induced to reproduce, and the extrusion occurred 200-255 degree-hours after hormonal induction at 24 degrees C. Larval ontogeny was evaluated every 10 minutes under microscopic/stereomicroscopic using fresh eggs samples. The main embryogenic development stages were identified: zygote, cleavage, including the morula, blastula, gastrula phase, organogenesis, and hatching. The extruded oocytes showed an average diameter of 1.10 +/- 0.10 mm, and after fertilization and hydration of eggs, the average diameter of eggs increased to about 1.90 +/- 0.60 mm, characterized by a large perivitelline space that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). Cell division started about 2 minutes after fertilization (AF), resulting in 2, 4, 8 (4 x 2 arrangement of cells), 16 (4 x 4), 32 (4 x 8) and 64 (2 x 4 x 8) cells. Furthermore, the blastula and gastrula stages followed after these cells divisions. The closed blastopore occurred at 11 h 20 min AF; following the development, the organogenetic stages were identified and subdivided respectively in: early segmentation phase and late segmentation phase. In the early segmentation phase, there was the establishment of the embryonic axis, and it was possible to distinguish between the cephalic and caudal regions; somites, and the optic vesicles developed about 20 h AF. Total hatching occurred at 54 h AF, and the larvae average length was 4.30 +/- 0.70 mm. Gradual yolk sac reduction was observed during the first two days of larval development. The first feeding occurred at the end of the second day. During the larval phase, cannibalism, heterogeneous larval growth and photophobia were also observed. This information will be important in improving the artificial reproduction protocols of S. parahybae in controlled breeding programs.

Fungal Transformation and Schistosomicidal Effects of Pimaradienoic Acid

The schistosomicidal effects of pimaradienoic acid (PA) and two derivatives, obtained by fungal transformation in the presence of Aspergillus ochraceus, were investigated. PA was the only compound with antischistosomal activity among the three diterpenes studied, with the ability to significantly reduce the viability of the parasites at concentrations ranging from 25 to 100 mu M. PA also promoted morphological alterations of the tegument of Schistosoma mansoni, separated all the worm couples, and affected the production and development of eggs. Moreover, this compound was devoid of toxicity toward human fibroblasts. In a preliminary in vivo experiment, PA at a dose of 100 mg/kg significantly diminished the number of parasites in infected Balb/c mice. Taken together, these results show that PA may be potentially employed in the discovery of novel schistosomicidal agents, and that diterpenes are an important class of natural compounds for the investigation of agents capable of fighting the parasite responsible for human schistosomiasis.

Costs and benefits of temporary brood desertion in a Neotropical harvestman (Arachnida: Opiliones)

Brood desertion is a life history strategy that allows parents to minimize costs related to parental care and increase their future fecundity. The harvestman Neosadocus maximus is an interesting model organism to study costs and benefits of temporary brood desertion because females abandon their clutches periodically and keep adding eggs to their clutches for some weeks. In this study, we tested if temporary brood desertion (a) imposes a cost to caring females by increasing the risk of egg predation and (b) offers a benefit to caring females by increasing fecundity as a result of increased foraging opportunities. With intensive field observations followed by a model selection approach, we showed that the proportion of consumed eggs was very low during the day and it was not influenced by the frequency of brood desertion. The proportion of consumed eggs was higher at night and it was negatively related to the frequency of brood desertion. However, frequent brood desertion did not result in higher fecundity, measured both as the number of eggs added to the current clutch and the probability of laying a second clutch over the course of the reproductive season. Considering that harvestmen are sensitive to dehydration, brood desertion during the day may attenuate the physiological stress of remaining exposed on the vegetation. Moreover, since brood desertion is higher during the day, when egg predation pressure is lower, caring females could be adjusting their maternal effort to the temporal variation in predation risk, which is regarded as the main cost of brood desertion in ectotherms.

Involvement of TSSA (trypomastigote small surface antigen) in Trypanosoma cruzi invasion of mammalian cells

TSSA (trypomastigote small surface antigen) is a polymorphic mucin-like molecule displayed on the surface of Trypanosoma cruzi trypomastigote forms. To evaluate its functional properties, we undertook comparative biochemical and genetic approaches on isoforms present in parasite stocks from extant evolutionary lineages (CL Brener and Sylvio X-10). We show that CL Brener TSSA, but not the Sylvio X-10 counterpart, exhibits dose-dependent and saturable binding towards non-macrophagic cell lines. This binding triggers Ca2+-based signalling responses in the target cell while providing an anchor for the invading parasite. Accordingly, exogenous addition of either TSSA-derived peptides or specific antibodies significantly inhibits invasion of CL Brener, but not Sylvio X-10, trypomastigotes. Non-infective epimastigote forms, which do not express detectable levels of TSSA, were stably transfected with TSSA cDNA from either parasite stock. Although both transfectants produced a surface-associated mucin-like TSSA product, epimastigotes expressing CL Brener TSSA showed a similar to 2-fold increase in their attachment to mammalian cells. Overall, these findings indicate that CL Brener TSSA functions as a parasite adhesin, engaging surface receptor(s) and inducing signalling pathways on the host cell as a prerequisite for parasite internalization. More importantly, the contrasting functional features of TSSA isoforms provide one appealing mechanism underlying the differential infectivity of T. cruzi stocks.

Production of Excirolana armata (Dana, 1853) (Isopoda, Cirolanidae) on an exposed sandy beach in southeastern Brazil

The somatic and gonad productions of the cirolanid isopod Excirolana armata were analyzed by taking monthly samples from December 2003 to November 2005 on Una beach, So Paulo state (24A degrees S), southeastern Brazil. Sampling was performed along three fixed transects established from the base of the foredunes to the waterline. Weight-specific growth rate was used to estimate the E. armata somatic production for 2004 and 2005, separately. The gonad production was estimated based on the monthly reproductive potential (mean number of eggs/embryos per female x monthly abundance of ovigerous females with near-release broods) for 2004. The annual somatic production of E. armata population varied from 15.57 to 17.25 g AFDW m(-1) year(-1) and the somatic production/biomass ratio (P (s)/B) from 3.55 to 3.14 year(-1) for 2004 and 2005, respectively. The P (s)/B ratios were higher for males (4.02 and 3.19 year(-1) for 2004 and 2005) than for females (3.10 year(-1) for both years). The annual gonad production (P (g) = 1.07 g AFDW m(-1) year(-1)) contributed about 15 and 6% to the total production (P (s) + P (g)) of females and the population, respectively. The proportion of gonad to somatic production of females (P (g)/P (s)) increased with individual size (ca 90% in the 7.5 mm size class), and the annual weight-specific gonad production (P (g)/B ratio) was estimated to 0.24 year(-1). The high P (s)/B ratios estimated for E. armata derive from the fast growth of individuals and show the importance of this population to the energy flow on Una beach ecosystem. However, the low percentage of juveniles verified in this population and in other studies of populations of the genus Excirolana is discussed as an important source of underestimation of P (s)/B ratio.