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ABSTRACT: Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs) mainly caused by roundworms (ascarids) and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides) and other predator (Duddingtonia flagrans). Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4 · 12 H2O, and wheat (Triticum aestivum)). A Fasciola hepatica recombinant protein (FhrAPS) was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor) and a predator species (Duddingtonia) for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs.
BioMed research international. 01/2013; 2013:567876.
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P Mula,
C Pilo,
C Solinas,
A P Pipia,
A Varcasia,
I Francisco, M S Arias,
A Paz Silva,
R Sánchez-Andrade,
P Morrondo,
P Díez-Baños,
A Scala
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ABSTRACT: From January to December 2008, 265 horses slaughtered in Sardinia (Italy) were examined for the presence of Rhinoestrus spp. (Diptera: Oestridae) through the examination of the nasal cavities and pharynges. Larvae were detected in 49% of the horses, with a mean intensity of infestation of 16.09 and abundance of 7.95. A total of 2108 larvae were collected, 66% of which were classified in first instar (L1), 22% in second instar (L2) and 12% in third instar (L3). The most frequent localization of larvae was the ethmoid, while the less one the larynx. According to the dynamics of Rhinoestrus larval stages, three periods in the chronobiology can be considered, the diapause (September-February) characterized by an absolute prevalence of first larval stage; the active phase of the endogenous phase (February-September) with an increase in the percentages of L2 and L3, and the exit phase (May-September), pointed by a further increase of L1. Morphological examination of L3 larvae revealed the presence of the Rhinoestrus purpureus features in 8% of the examined larvae, of 8% of the Rhinoestrus usbekistanicus features, while in 84% of the larvae were evidenced intermediate features. Contrastingly biomolecular analysis of the COI gene of the larvae evidenced uniformity at genetic level, confirming the presence of a unique species in the Mediterranean area. The results of the present paper, reveal the wide diffusion of rhinoestrosis among Sardinian horses, and suggest the need for applying appropriate control measures. Chemotherapy should be very useful if administered during the diapause period, for reducing the presence of L1 stages and interrupting thus the life cycle of this myiasis.
Veterinary Parasitology 10/2012; · 2.58 Impact Factor
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ABSTRACT: The analysis of infection by Paramphistomidae trematodes was conducted in two agricultural regions with different knowledge on this parasitosis. Faecal and blood samples were collected from 374 cattle in Salto (NW Uruguay) where there is a lack of information about paramphistomosis. A total of 429 cattle from Galicia (NW Spain), an area with previous records of infection by gastric flukes, were sampled. Diagnostics of trematodosis was developed by using a copromicroscopic probe and an ELISA with excretory/secretory antigens collected from adult Calicophoron daubneyi (Paramphistomidae) specimens. Results were evaluated according intrinsic and extrinsic factors. In the Uruguay, the percentage of cattle passing Paramphistomidae-eggs by faeces was 7% (95% Confidence Interval 5, 10). A significantly higher prevalence of paramphistomosis in the Hereford×Angus cattle (OR=3.5) was recorded, as observed for the oldest ruminants (>3.5years). An overall seroprevalence of 29% (25, 34) was obtained by ELISA, with the highest values in the Friesians (OR=3), the youngest bovines (<2.5years) and dairy cattle (Friesians). Twenty-six percent (22, 30) of the cattle from Spain passed eggs by faeces, and cattle aged 2.5-7years reached significant highest prevalences. By means of the ELISA, a percentage of 55 cattle (50, 59) had antibodies against the gastric fluke, and the highest seroprevalence was observed among the bovines under 6years. It is concluded that paramphistomosis is on the increase in cattle from NW Spain, partly due to the absence of an effective treatment against the trematode. There is a need for reducing the risk of infection by Paramhistomidae spp. in cattle from Uruguay, especially by improving their management to avoid exposure to the gastric trematode. Further studies are in progress for identifying the species of Paramphistomidae affecting ruminants in Uruguay.
Veterinary Parasitology 08/2012; · 2.58 Impact Factor
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ABSTRACT: Three major protein complexes (51, 29, and 15 kDa, named P1 to P3, respectively) were resolved by gel filtration of the excretory/secretory antigens collected from a mixture of horse cyathostomin third-stage larvae (L3s). The potential application for the detection of infected horses was assessed with an enzyme-linked immunosorbent assay (ELISA) by the comparison of the serological and copromicroscopical results. The value of the area under the receiver operating characteristic (ROC) curve was higher than 0.9 when the three peaks were used. Elevated values (>90%) for the sensitivity, specificity, and the positive-likelihood ratio were also observed for all the antigen complexes. A significant increment in the IgG antibody levels 4 weeks prior to the observation of eggs in the feces of weanlings naturally infected was recorded. Our results indicate that the evaluation of chemotherapy is possible by using immunoenzymatic probes and fast protein liquid chromatography (FPLC)-purified antigens. Data collected in the present investigation indicate that FPLC isolation offers a very helpful one-step method for collecting antigens with diagnostic potential to be employed in immunoenzymatic probes.
Clinical and vaccine immunology: CVI 07/2011; 18(9):1462-6. · 2.37 Impact Factor
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R Sánchez-Andrade,
F J Cortiñas,
I Francisco,
J A Sánchez,
P Mula,
C Cazapal,
L Vázquez,
J L Suárez,
R Francisco, M S Arias,
P Díez-Baños,
A Scala,
A Paz-Silva
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ABSTRACT: We have developed a novel enzyme-linked immunosorbent assay (ELISA) based on excretory/secretory antigens of second instar Gasterophilus for the diagnosis of gasterophilosis in grazing horses. Between January 2007 and January 2009, two experiments were carried out on free-ranging horses in northwest Spain. During the first year, monthly blood samples were collected from a herd of 25 horses. In the second year, a monthly serological survey was conducted for a total of 398 different horses. All the sera were analyzed by ELISA using excretory/secretory antigens from Gasterophilus intestinalis (GphiL2ES) and Gasterophilus nasalis second-stage larvae (GphnL2ES). Climatic data were collected between January 2007 and January 2009 from local meteorological automated stations to establish the weather pattern in the study area. Observations of Gasterophilus eggs on the horses' hair and third instars passed in the faeces were also done. The kinetics of IgG response decreased against GphiL2ES from January to July, increased slowly from August and rose up to January. After a slight decrease in January, the absorbances against GphnL2ES reduced from April to August, when the lowest values were observed. The IgG values rose until the end of the study in January. Third instars were observed in the faeces in March to May, and Gasterophilus eggs were seen on the horses' hair from June to September. The highest IgG seroprevalences were achieved in winter (January-February; 100%) against both antigens. The lowest percentages of seropositivity were observed in June (3%) to the GphiL2ES, and in July (9%) to the GphnL2ES. The use of antigens from G. intestinalis second-stage larvae was shown to be suitable for diagnosing infestation by G. intestinalis or G. nasalis. We concluded that under oceanic climate conditions, the egg-laying period occurs from late spring, and eggs and first instars are found in the mouth in early summer. During summer the second instars move into the stomach and intestine, where the third-stage larvae remain until the end of winter, when pupation takes place. The adult horse bot fly emerges in the spring. Two treatments for the control of gasterophilosis are suggested: a curative in the summer to eliminate the first instars and a preventive in the autumn to suppress the second instars.
Veterinary Parasitology 08/2010; 171(3-4):314-20. · 2.58 Impact Factor
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J A Romero, M S Arias,
J L Suárez,
A Paz-Silva,
I Francisco,
F Alonso,
F J Cortiñas,
V Dacal,
A Romasanta,
P Morrondo,
P Díez-Baños,
A Scala,
R Sánchez-Andrade
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ABSTRACT: The analysis of immunoglobulin (Ig) M and IgG serum isotypes as indicators of the presence of Oestrus ovis developmental stages was carried out. A serological survey to discover the seasonal variations in the prevalence of oestrosis in sheep from an oceanic climate area was developed. Six hundred and sixty-nine blood samples were analyzed for the presence of IgM and IgG using an enzyme-linked immunosorbent assay and O. ovis second-stage larvae excretory/secretory antigens. In addition, the effect of an ivermectin-based treatment on the humoral immune response was measured. The percentage of positive animals was 54% for IgM and 55% for IgG. The highest percentages of sheep positive to IgM were recorded in the winter and to IgG in the summer. No animals positive to IgM were detected in the ivermectin-treated sheep. These results seem to underline that oestrosis is a parasitic disease to keep watch for in regions with an oceanic climate. A noninvasive evaluation of the success of chemotherapy seems possible by analyzing the serum IgM response.
Journal of Medical Entomology 05/2010; 47(3):477-81. · 1.76 Impact Factor
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ABSTRACT: A series of 10α-acyloxy-N-phenethyl-8-azabicyclo[4.3.1]decane derivatives have been synthesized and studied by 1H and 13C nmr spectroscopy, and the crystal structure of N-phenethyl-10aL-(xanten-9″-carbonyloxy)bicyclo[4.3.1]decane 2 has been determined by X-ray diffraction. The compounds studied display in deuteriochloroform the same preferred conformation adopted by the piperidine ring as a distorted chair conformation flattened at N-8 with both the phenethyl and acyloxy groups in the equatorial position with respect to the piperidine ring. These results are in close agreement with that found for compound 2 in the crystalline state.
Journal of Heterocyclic Chemistry 03/2009; 26(4):965 - 970. · 1.22 Impact Factor
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ABSTRACT: A series of N-phenethyl-8-β-amidocamphidines 4a-f (3-phenethyl-8-β-(N-arylamido)-3-azabicyclo-[3.2.1]octane) has been designed, synthesized and stereochemically characterized as semirigid analogous of the 4-anilidopiperidine analgesics in an attempt to study the influence of certain stereochemical factors on analgesia in this class of compounds. In deuteriochloroform and deuteriobenzene solution, compounds 4a-f display the same preferred conformation. The cyclopentane and piperidine rings adopt an envelope and distorted chair conformation respectively flattened at N-3, with the N and C-8 substituents in equatorial and axial positions with respect to the piperidine ring.In vivo pharmacological testing demonstrated that compounds 4a-f were inactive in the analgesic test, with the exception of compound 4f which showed an ED50 of 250 mg/kg p.o.
Journal of Heterocyclic Chemistry 03/2009; 31(2):313 - 318. · 1.22 Impact Factor
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The Journal of Antibiotics 08/1996; 49(7):700-2. · 1.65 Impact Factor
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ABSTRACT: A series of 3 beta-acyloxy-3 alpha-methoxycarbonyltropanes were synthesized and studied by 1H and 13C NMR spectroscopy, and the crystal structure of 3 alpha-methoxycarbonyl-3 beta-pyridincarbonyloxytropane (5d) was determined by X-ray diffraction. In CDCl3 solution, compounds 5a-f display the same preferred conformation. The pyrrolidine and piperidine rings adopt an envelope conformation flattened at N-8 and a distorted chair conformation puckered at N-8 and flattened at C3, respectively, with the N-substituent in the equatorial position with respect to the piperidine ring. The pharmacological profile of one of these compounds makes it an adequate candidate for the design of novel GABAB antagonist agents.
Journal of Pharmaceutical Sciences 09/1993; 82(8):794-8. · 3.06 Impact Factor
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ABSTRACT: Two new pyrazolo-isoquinolinone antibiotics, APHE-1 and APHE-2, have been isolated from the culture filtrate and mycelia of Streptoverticillium griseocarneum NCIMB 40447. Molecular formulae were established as C13H12N2O for APHE-1 and C14H14N2O for APHE-2, by elemental analysis, NMR and mass spectra. 2D NMR techniques (1H-1H COSY-45 and 1H-13C correlated spectroscopy) have been applied to establish their structures.
The Journal of Antibiotics 12/1992; 45(11):1759-62. · 1.65 Impact Factor
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ABSTRACT: A series of 8-beta-acyloxy-3-phenethyl-3-azabicyclo[3.2.1]octane and its N-endo methiodides were synthesized and studied by 1H and 13C NMR spectroscopy, and the crystal structure of 8-beta-p-chlorobenzoyloxy-3-phenethyl-3-azabicyclo[3.2.1]octane methiodide (2c) was determined by X-ray diffraction. In CDCl3 solution, 1b-1e display the same preferred conformation. The cyclopentane and piperidine rings adopt an envelope conformation flattened at C-8 and a distorted chair conformation puckered at C-8 and flattened at N-3, respectively, with the N-substituent in the equatorial position with respect to the piperidine ring. In all cases, methylation takes place from the endo position. The ability of the title compounds to antagonize the acetylcholine-induced contraction of guinea pig ileum is also reported. An initial structure-activity relationship is proposed.
Journal of Pharmaceutical Sciences 07/1991; 80(6):554-8. · 3.06 Impact Factor
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ABSTRACT: The infrared and 1H and 13C NMR spectra of 3-aza-bicyclo[3.2.1]octane-8-β-ol have been examined in several media. To assist in interpretation of the spectroscopic data, the crystal structure has been determined by X-ray diffraction.The bicyclic system adopts a chair—envelope conformation with OH and phenethyl groups, respectively, in axial and equatorial positions with respect to the piperidine ring. The crystal structure is stabilized by means of OH…N intermolecular hydrogen bonding. In CCl4 solution the initial chair—envelope conformation changes to a boat—envelope conformation which is stabilized by an intramolecular hydrogen bond.The unambiguous assignment of all protons of the bicyclic system, not previously described, has also been carried out.
Journal of Molecular Structure.
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ABSTRACT: The infrared, 1H and 13C NMR spectra of 9-(2′-hydroxyethyl)-9-azabicyclo[3.3.1] nonan-3α-ol (I) have been examined in several media. The crystal structure has been determined by X-ray diffraction. The bicycle system adopts in solid state a distorted double chair conformation, being the OH and 2′-hydroxyethyl group attached in axial position with respect to the piperidinol ring. In solution, by the contrary, the title compound adopts a preferred flattened chair-boat conformation with the piperidinol ring in a slight distorted boat form. The unambiguous assignment of all protons of the granatanine system, not described up to date, has been carried out.
Journal of Molecular Structure. 174:273-280.
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ABSTRACT: The conformational preferences of α(I) and β(II) epimers of 3-phenethyl-3-azabicyclo[3.3.1]nonan-9-ol, have been studied using the molecular mechanics method (MM2 and MM2′ programs). In both cases, the global minimum-energy conformer computed correspond to a slightly flattened CC conformation with the 3N-phenethyl group in the β-position. This behaviour is in agreement with the conformation adopted in solution in polar solvents. In relation to the energy differences between N-axial and N-equatorial conformations, these epimers exhibit an intermediate situation between the tropine and the α and β epimers of 3-phenethyl-3-azabicyclo[3.2.1]octan-8-ol.
Journal of Molecular Structure. 266:259-264.
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ABSTRACT: A series of 8-β-alcoxycarbonyl-8-α-hydroxy-3-phenethyl-3-azabicyclo[3.2.1]octane derivatives have been synthesized and studied by IR,1H and 13C NMR spectroscopy, and the crystal structure of ethyl-8-α-hydroxy-3-azabicyclo[3.2.1]octane-8-β-carboxylate (VI) has been determined by X-ray diffraction. The compounds studied display in deuterochloroform and benzene-d6 the same preferred chair-envelope conformation flattened at N-8 and puckered at C-3 with both the phenethyl and hydroxy groups in the equatorial position with respect to the piperidine ring. These results are in close agreement with those found for compound VI in the crystalline state. By comparing the NMR parameters of these compounds with those of 3-phenethyl-3-azabicyclo [3.2.1]octan-8-α-ol (VII) several stereoelectronic effects have been deduced.
Journal of Molecular Structure.
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ABSTRACT: The conformational preferences of the 3-azabicyclo[3.3.1]nonan-9β-ol (I) and several 3-alkylderivatives have been studied by means of the molecular mechanics method (MM2, MM2′ and MMX programs). In all cases, the most stable conformer corresponds to a slightly flattened chair-chair conformation with the 3N-substituent in the β-disposition. According to the theoretical values: 1) the steric hindrance of the 3N-group is not very important in determining the energy differences between conformers; 2) the conformational inversion of the 3N-substituent is favoured by considering intramolecular hydrogen bond.
Journal of Molecular Structure. 293:43-48.
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ABSTRACT: N-Substituted 9-azabicyclo [3.3.1] nonan-3-ones have been studied by 1H- and 13C-NMR spectroscopy. In CDCl3 solutions, these ketones can be described through an equilibrium between two flattened chair-chair conformers by nitrogen inversion. The piperidone ring is more flattened than the piperidine one and has a slight preference for the form with the N-substituent axial to the piperidone ring. The contribution of the conformers with piperidone or piperidine rings in an ideal boat conformation must be low. The application of the Altona empirical equation supports this hypothesis.
Journal of Molecular Structure.
