Authentication of the botanical and geographical origin of honey by front-face fluorescence spectroscopy

Front-face fluorescence spectroscopy, directly applied on honey samples, was used for the authentication of 11 unifloral and polyfloral honey types (n = 371 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis. Excitation spectra (220-400 nm) were recorded with the emission measured at 420 nm. In addition, emission spectra were recorded between 290 and 500 nm (excitation at 270 nm) as well as between 330 and 550 nm (excitation at 310 nm). A total of four different spectral data sets were considered for data analysis. Chemometric evaluation of the spectra included principal component analysis and linear discriminant analysis; the error rates of the discriminant models were calculated by using Bayes' theorem. They ranged from <0.1% (polyfloral and chestnut honeys) to 9.9% (fir honeydew honey) by using single spectral data sets and from <0.1% (metcalfa honeydew, polyfloral, and chestnut honeys) to 7.5% (lime honey) by combining two data sets. This study indicates that front-face fluorescence spectroscopy is a promising technique for the authentication of the botanical origin of honey and may also be useful for the determination of the geographical origin within the same unifloral honey type.     

Efficacy of albendazole and moxidectin and resistance to ivermectin against Libyostrongylus douglassii and Libyostrongylus dentatus in ostriches

Anthelmintic resistance has emerged globally as a problem amongst nematode of livestock and has been particularly well documented in equine and small ruminants. There are no studies regarding the efficacy of anthelmintics against the hematophagous nematodes in ostriches, Libyostrongylus dentatus; and just a few on L. douglassii. Here the efficacy of albendazole, ivermectin and moxidectin were evaluated against these two species in an ostrich farm in Minas Gerais state, Brazil. The feces were collected on the day of treatment and after 13days of an oral dose of albendazole (6mg/kg), or an injected dose (0.2mg/kg) of ivermectin or moxidectin. The fecal egg count reduction test and coprocultures were performed to determine possible resistance against the drugs used. An efficacy of 60% was found for ivermectin, while albendazole and moxidectin were 100% effective. Both worm species appeared to have reduced sensitivity to ivermectin.     

Secretion of multi-protein migratory complex induced by Toxoplasma gondii infection in macrophages involves the uPA/uPAR activation system

Toxoplasmosis is a world wide spread zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite that is able to disseminate into deep tissues and cross biological barriers, reaching immunoprivileged sites such as the brain and retina. The parasite is able to infect macrophages and dendritic cells for dispersal throughout the body. However, the molecular mechanisms or outcomes of the subversion of the host cell are largely unknown. Recently our group established that metalloproteinases are involved in migration of infected macrophages. Herein, we evaluated the recruitment of host invasive machinery components in T. gondii infected murine macrophages. We showed by immunoprecipitation assays that MMP-9, CD44 TIMP-1 and uPAR were secreted as a multi-protein complex by infected macrophages. Zymographic analysis revealed that MMP-9 was present in its pro- and active form. Moreover, inhibition of uPA/uPAR pathway by PAI-1 decreased secretion of MMP-9 active forms, as well those associated to uPAR and TIMP-1, but not to CD44. Data presented here suggest that MMP-9 is secreted as a multiprotein complex by T. gondii infected macrophages, similar to that observed in metastatic cells. We further speculate that uPA/uPAR system is involved in the expression/secretion of complexes containing active MMP-9 forms.     

A new type of pterocarpanquinone that affects Toxoplasma gondii tachyzoites in vitro

Toxoplasma gondii, the agent of Toxoplasmosis, is an obligate intracellular protozoan able to infect a wide range of vertebrate cells, including nonprofessional and professional phagocytes. Therefore, drugs must have intracellular activities in order to control this parasite. The most common therapy for Toxoplasmosis is the combination of sulfadiazine and pyrimethamine. This treatment is associated with adverse reactions, thus, the development of new drugs is necessary. In previous studies, naphthoquinone derivatives showed anti-cancer activity functioning as agents capable of acting on groups of DNA, preventing cancer cells duplication. These derivatives also display anti-parasitic activity against Plasmodium falciparum and Leishmania amazonensis. The derivative pterocarpanquinone tested in this work resulted from the molecular hybridization between pterocarpans and naphtoquinone that presents anti-tumoral and anti-parasitic activities of lapachol. The aim of this work was to determine if this derivative is able to change T. gondii growth within LLC-MK2 cells. The drug did not arrest host cell growth, but was able to decrease the infection index of T. gondii with an IC(50) of 2.5 μM. Scanning and transmission electron microscopy analysis showed morphological changes of parasites including membrane damage. The parasite that survived tended to encyst as seen by Dolichos biflorus lectin staining and Bag-1 expression. These results suggest that pterocarpanquinones are drugs potentially important for the killing and encystment of T. gondii.     

The virB-encoded type IV secretion system is critical for establishment of infection and persistence of Brucella ovis infection in mice

Brucella spp. are gram-negative intracellular bacterial pathogens that cause chronic infections. Brucella virulence factors include a type IV secretion system (T4SS) and its lipopolysaccharide (LPS), which are essential for persistence. However, the role of the virB-encoded T4SS has not been investigated in naturally rough Brucella species such as Brucella ovis. In this study, male 6-week old BALBc mice were infected with B. ovis, Brucella abortus, and their respective ΔvirB2 mutant strains. During early infection, B. ovis and B. abortus wild type strains were similarly recovered from spleen. Interestingly, in contrast to ΔvirB2 B. abortus that was recovered at similar levels when compared to the wild type strain, the ΔvirB2 B. ovis was markedly attenuated as early as 24h post infection (hpi). The ΔvirB2 B. ovis was unable to survive and multiply in murine peritoneal macrophages and extracellularly within the peritoneal cavity at 12 and 24 hpi with lower splenic colonization than the parental strain at 6, 12 and 24 hpi. In contrast, wild type B. abortus and ΔvirB2 B. abortus had a similar kinetics of infection in this model. As expected, the T4SS was essential for intracellular replication of smooth and rough strains in RAW macrophages at 48 hpi. These results suggest that T4SS is important for survival of B. ovis in murine model, and that a T4SS deficient B. ovis strain is cleared at earlier stages of infection when compared to a similar B. abortus mutant.     

Mydriatic effect of topically applied rocuronium bromide in tawny owls (Strix aluco): comparison between two protocols

Objective To evaluate the mydriatic efficacy of a neuromuscular blocking agent (rocuronium bromide) applied topically to only one eye of nocturnal birds of prey and to assess for any general and/or local adverse effects due to its use. Animal studied Twelve healthy adult tawny owls (Strix aluco) were randomly divided in two groups. Procedures Six birds (Group 1) received a single dose of 0.35 mg of rocuronium bromide. The second group of subjects (Group 2) received two doses of 0.35 mg of rocuronium bromide (total 0.70 mg/eye). In both groups, the curariform agent was instilled topically. Pupil diameter was measured with a pupillary gauge in 10 min intervals for a total of 100 min and then every 20 min for a total of 240 min. The assessment of the pupillary light reflex was performed using a standard light source during pupillary size recording. Results Maximal pupillary diameter was 11.5 ± 0.3 mm for Group 1 and 11.0 ± 0.6 mm for Group 2 and no statistically significant differences were detected among the two groups. The maximal pupillary diameter was achieved at T80 for Group 1, and at T60 for Group 2. A complete fundus examination was possible on all treated eyes of subjects of both groups. The drug did not cause any noticeable adverse effects in any of the examined birds. Conclusion Results of the present study suggest that a single topical administration of 0.35 mg of rocuronium bromide to the eyes of healthy tawny owls results in sufficient mydriasis to allow for a complete examination of the fundus.     

Evidence of farm-level synergies in mixed-farming systems in the Australian Wheat-Sheep Zone

Synergies are reported for mixed-enterprise farming systems in a benchmarking group in the Wheat-Sheep Zone in New South Wales, Australia. A stochastic input distance function is estimated and a measure of synergies is calculated, based on the second cross partial derivative of output variables in the distance function. Evidence is presented of strong synergies from complementarity between sheep and beef enterprises, sheep and crop enterprises, and beef and crop enterprises.     

Deduced Amino Acid Sequence of an α-Gliadin Gene from Spelt Wheat (Spelta) Includes Sequences Active in Celiac Disease

The complete amino acid sequence of an α-type gliadin from spelt wheat (spelta) has been deduced from the cloned DNA sequence and compared with α-type gliadin sequences from bread wheat. The comparison showed only minor differences in amino acid sequences between the α-type gliadin from bread wheat and the α-type gliadin from spelta. The two sequences had an identity of 98.5%. Larger differences can be found between different α-type gliadin amino acid sequences from common bread wheat. Because all the different classes of gliadins, α, β, γ, and ω, appear to be active in celiac disease, it is reasonably certain that the spelta gliadin is also toxic. We conclude that spelta is not a safe grain for people with celiac disease, contrary to the implications in labeling a bread made from spelta as “an alternative to wheat”. Our conclusions are in accord with spelta and bread wheat being classed taxonomically as subspecies of the same genus and species, Triticum aestivum L.