Seroprevalence of and risk factors for Toxoplasma gondii in sheep raised in the Jaboticabal microregion, São Paulo State, Brazil

Seroprevalence of and risk factors for toxoplasmosis in sheep from different properties in the Jaboticabal microregion, São Paulo State, Brazil were determined. Antibodies to Toxoplasma gondii were found in sera of 52.0% of 488 sheep tested by indirect fluorescent antibody test (IFAT ? 64). T. gondii seropositivity in sheep was significantly associated with gender of the sheep, pasturing system, contact with cats, and the use of mineral supplements and the type of feed.     

Multiplex RT-PCR for detection and identification of three necroviruses that infect olive trees

An optimized multiplex RT-PCR assay was developed to discriminate three necrovirus (Olive latent virus 1 (OLV-1), Tobacco necrosis virus D (TNV-D) and Olive mild mosaic virus (OMMV)) that infect olive trees. An olive orchard consisting of 54 trees of cv. “Galega vulgar” in the south of Portugal was surveyed. dsRNA fraction was used as template and revealed the 3 viruses, singly or in multiple infections, present in 17 out of 54 trees in the orchard. OMMV was the most frequent occurring in 15 trees, followed by OLV-1 in 12 and TNV-D in 4 plants. The results obtained showed that necrovirus- specific dsRNAs do exist in infected tissues in amounts below the resolution permitted by gel electrophoresis analysis and that the developed multiplex PCR based assay is of much higher sensitivity. The design of the specific primers described enabled, for the first time, to discriminate between OMMV and TNV-D by means of RT-PCR assays, an indispensable tool in identification, epidemiology and survey studies.     

Babesia canis canis and Babesia canis vogeli infections in dogs from northern Portugal

Canine babesiosis represents an important veterinary medical problem. This study describes the molecular characterization of babesial parasites detected in eight clinically suspected dogs from northern Portugal, affected by lethargy, muscle tremors, weight loss, pale mucous membranes, hyperthermia or red-coloured urine. Microscopic examination of peripheral blood smears showed large intraerythrocytic piroplasms morphologically compatible with Babesia canis in all eight animals. DNA was extracted from blood on filter paper, and a Babesia spp. infection confirmed by polymerase chain reaction (PCR) amplification of a 408bp fragment of the 18S rRNA gene. Analysis of PCR-derived sequences revealed that seven dogs were infected with B. canis canis and one with B. canis vogeli. This is the first molecular identification report of both the species B. canis and the subspecies B. canis canis and B. canis vogeli in dogs from Portugal.     

Serological evaluation of experimentally infected dogs by LicTXNPx-ELISA and amastigote-flow cytometry

Canine visceral leishmaniasis (CVL) is characterized by a high incidence of asymptomatic infections. Because of the high prevalence of asymptomatic dogs in the endemic areas of visceral leishmaniasis (VL), a sensitive test is required for an accurate diagnosis. In this study, we evaluated the detection of symptomatic and asymptomatic Leishmania infantum infection in dogs using the secreted LicTXNPx antigen (Leishmania infantum cytosolic tryparedoxin peroxidase) in an ELISA format and compared it to soluble Leishmania antigens from promastigote or amastigote forms (SPLA and SALA) and two other unrelated secreted Leishmania proteins (LiTXN1 and TDR1). Moreover, we evaluated the diagnostic potential using the promastigote or amastigote-flow cytometric methodologies. The assays utilized sera collected from a cohort of L. infantum experimentally infected dogs, in which the intravenous or intradermal parasite injection mimics a symptomatic or asymptomatic pattern of infection, respectively. Our study indicated that anti-LicTXNPx antibodies were present in both symptomatic and asymptomatic experimental infections. Among the different Leishmania recombinant proteins tested, LicTXNPx showed a good predictive correlation with total soluble promastigote or amastigote Leishmania antigens, suggesting this antigen as a good candidate for a marker in either symptomatic or asymptomatic infection. The use of flow cytometry using both forms of live parasites was also tested with the same group of dogs. Amastigotes were shown to have more advantages than promastigotes for the serological diagnostic in both symptomatic and asymptomatic dogs, since higher continuous levels of anti-amastigote antibodies were detected during the course of experimental infection. Moreover, additional studies were done using sera from non-infected dogs and clinically asymptomatic and symptomatic dogs with confirmed naturally occurring L. infantum infections. The sensitivities of amastigote and promastigote flow cytometry were 96% vs. 89%, respectively, while the specificity for both was 93.2%. Therefore, our findings showed for the first time the potential of amastigote-flow cytometry regarding their applicability to detect both symptomatic and asymptomatic VL canine infections.     

Productivity improvement of Lysmata seticaudata (Risso, 1816) larval rearing protocol through modelling

The Monaco shrimp Lysmata seticaudata (Risso, 1816) is a marine ornamental species whose ecology and biology, as well as its larval culture has previously been addressed. The objective of the study was to predict and improve productivity of this species rearing protocol through modelling. The models developed intend to help aquaculturists to maximize survival to postlarva, decrease larval duration and increase synchronism of metamorphosis and newly metamorphosed postlarvae size by manipulating temperature, diet, first feeding period and stocking density.The models developed allow us to conclude that the L. seticaudata rearing protocol productivity can be improved by raising larvae at a density of 40 larvae L^− 1 and fed newly hatched Artemia nauplii since hatching to zoea V, and with Algamac 2000™ enriched Artemia metanauplii from zoea V to metamorphosis to postlarvae.By providing more productive protocols to aquaculturists, destructive practices and wild collection may be reduced.     

Incorporation of trace elements on iron-rich concretions around plant roots of tagus estuary salt marsh (portugal)

Objective

The concentration gradients of solid Al, Fe, Mn, Zn, Cd, Pb, Cr and Ni in freshly formed concretions and in older concretions preserved in the sediment were evaluated in a millimetre resolution scale. These results provide a better understanding of the complex biogeochemical processes in the root-sediment system and elucidate the potentialities of marshes in the restoration of contaminated aquatic systems.

Methods

Tens (soll das vielleicht tonnes/tons heißen) of freshly formed concretions and preserved concretions were collected in the Rosário salt marsh. Each tubular structure and involving sediments were scraped in concentric layers of 2-mm thickness to form two sets of composite samples. The elemental composition of concretion and sediment samples were determined by atomic absorption spectrometry.

Results and Discussion

The sediment involving the concretions contained ～0.5 mmol g^-1 Fe, but concentrations increased towards the root up to 3.4 mmol g^-1 in 3-mm interval. Manganese, Zn, Cd and Pb exhibited similar radial distribution in both preserved and freshly formed concretions indicating that these metals remain in those structures with the age. Chromium and Ni showed a different pattern being lower in concretions with respect to involving sediments, and consequently, concretions do not act as a barrier for these elements. The estimated amount of Zn (59 tons), Pb (5 tons) and Cd (0.4 tons) retained in the concretions all over the Rosario salt marsh highlights the importance of these structures in sequestering metals.

Conclusions and Outlook

The sequestering may be particularly relevant in salt marshes located near urban and industrial zones, being areas working as a natural remediation system and contributing for the detoxification of aquatic environments and the well-being of coastal resources.

     

Erylusamides: Novel Atypical Glycolipids from Erylus cf. deficiens

Among marine organisms, sponges are the richest sources of pharmacologically-active compounds. Stemming from a previous lead discovery program that gathered a comprehensive library of organic extracts of marine sponges from the off-shore region of Portugal, crude extracts of Erylus cf. deficiens collected in the Gorringe Bank (Atlantic Ocean) were tested in the innovative high throughput screening (HTS) assay for inhibitors of indoleamine 2,3-dioxygenase (IDO) and showed activity. Bioassay guided fractionation of the dichloromethane extract led to the isolation of four new glycolipids, named erylusamide A–D. The structures of the isolated compounds were established by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy, high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and chemical derivatization. The metabolites shared a pentasaccharide moiety constituted by unusual highly acetylated d-glucose moieties as well as d-xylose and d-galactose. The aglycones were unprecedented long chain dihydroxyketo amides. Erylusamides A, B and D differ in the length of the hydrocarbon chain, while erylusamide C is a structural isomer of erylusamide B.     

Formation of hemoglobin adducts of acrylamide after its ingestion in rats is dependent on age and sex

The effect of fiber and fat contents of food and of age and sex of animals on the formation of hemoglobin adducts of acrylamide (AA-Hb) in blood has been studied. The results suggest that the absorption of acrylamide (AA) present in food is not affected by the fiber or fat contents of food. However, AA-Hb resulting from the intake of an aqueous solution of AA is dependent on the age and sex of rats: AA-Hb levels were higher in females than in males (3.53- and 2.55-fold higher, respectively, for AA doses of 25 and 100 mg/kg) and in younger than in older rats (30.1% higher in 1.5 month old as compared to 14 month old rats). In males, AA-Hb levels found after the oral administration of AA in an aqueous solution were significantly lower than those found after dietary or intravenous administration. In conclusion, these results show the existence of significant differences in AA bioavailability from an aqueous solution depending on the sex and age of animals. If similar differences also occur in humans, they would be relevant to assess the exposure of different subpopulations to AA.     

Color properties of four cyanidin-pyruvic acid adducts

Four anthocyanin-pyruvic adducts were synthesized through the reaction of cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-sophoroside, and cyanidin 3-O-sambubioside with pyruvic acid, structurally characterized by liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR), and their chromatic properties were studied (pH and SO2 stability assays). Overall, these pigments were shown to have a higher resistance to discoloration toward pH variations and also in the presence of SO2, being that this resistance to discoloration was explained by a higher protection of the chromophore group against the water or bisulfite nucleophilic attack that gives rise to the colorless hemiacetal form. Only slight differences in the protection against the nucleophilic attack of water and bisulfite were found to occur between all of the cyanidin-pyruvic acid adducts studied. Indeed, anthocyanin-pyruvic acid adducts with glucose or sambubiose attached to the 3-O position of the flavylium moiety were shown to have smaller bleaching constants compared with similar pigments that possess a rutinosyl or sophorosyl moiety. The study of the pigments (A-D and cyanidin-3-O-glucoside) color parameters, namely, chroma (C), lightness (L), and the hue angle (h(a,b)), obtained from the CIELAB system, revealed that different patterns of sugars in the anthocyanin-pyruvic acid adduct moiety affected the referred three parameters of color. The loss of saturation (DeltaC < 0) and the increase of lightness (DeltaL > 0) presented by the cyanidin-pyruvic acid adduct solutions at acidic pH values (1.0 and 2.0) showed that they are much less colored than the cyanidin-3-O-glucoside. For higher pH values (5.0 and 7.0), the reverse trend was observed. This means that the cyanidin-pyruvic acid adducts A-D are much more colored than the anthocyanin at these pH values. The higher coloring capacity of these pigments at higher pH values may be an important feature, indicating a putative application of these compounds in food products.