Characterization and antioxidant potential of Brazilian fruits from the Myrtaceae family

The objective of this study is to evaluating the Brazilian biodiversity through physicochemical characterization and determination of antioxidant potential of three species from the Myrtaceae family, namely yellow guava (Psidium cattleyanum Sabine), guabiroba (Campomanesia xanthocarpa O. Berg), and uvaia ( Eugenia pyriformis Cambess). Guabiroba had the greater quantity of phenolic compounds (9033 mg chlorogenic acid/100 g) and vitamin C (30.58 mg/g) and showed the best TSS/TTA (total soluble solid/total titratable acid) ratio (45.12). For the ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic) method, the guabiroba (507.49 μM Trolox/g) presented the highest antioxidant potential; however, in the DPPH (2,2-diphenyl-1-picrylhydrazyl) method, uvaia (170.26 g/g DPPH) and guabiroba (161.29 g/g DPPH) were not statistically different. The uvaia outranked the other fruits with respect to its high carotenoid (909.33 μg/g) and vitamin A (37.83 μg/g) contents, and the yellow guava, although showing a lower bioactive compound content and antioxidant activity, nevertheless presented much higher values than many traditionally consumed fruits.     

Seeding and mulching + seeding effects on post‐fire runoff,soil erosion and species diversity in Galicia (NW Spain)

The effects of two different soil rehabilitation treatments on runoff, infiltration, erosion and species diversity were evaluated in a shrubland area in Galicia (NW Spain) after an experimental fire by means of rainfall simulations. The treatments compared were: seeding, seeding + mulching and control (untreated). Rainfall simulations were conducted 9 months after fire and the application of soil rehabilitation treatments. A rainfall rate of 67 mm h⁻¹ was applied for 30 min to each runoff plot. Seeding significantly increased plant species richness in the treated plots relative to the control plots, although it had no effect on diversity or evenness. Rehabilitation treatments did not significantly increase soil cover or affect runoff and infiltration. Soil losses were low in all cases, varying from 75·6 kg ha⁻¹ in the seeded + mulched plots to 212·1 kg ha⁻¹ in the untreated plots. However, there were no significant differences in sediment yields between treatments. The percentage of bare soil appeared to be a critical variable in controlling runoff and erosion. Copyright © 2010 John Wiley & Sons, Ltd.     

Transfer, viability and colonisation of Campylobacter jejuni in the chicken vitellus and in embryos

1. The objective of this study was to evaluate the ability of Campylobacter jejuni to penetrate and colonise eggs from specific-pathogen-free (SPF) and heavy breeder hens, and to determine its effects on the viability of SPF embryos.

2. We detected C. jejuni in 10% of breeder hens and 20% of SPF eggs, which demonstrates the ability of the bacteria to go through the pores of eggs and contaminate the vitellus after 3?h of contact. These results indicate that there is a risk of contamination under commercial production conditions, where, after oviposition, there is contact between the egg and organic material such as faeces and blood.

3. We observed that in 80% of SPF eggs analysed, C. jejuni survived the 21-d incubation period. This positive result suggests that this microorganism was also responsible for early embryonic mortality.

4. The ability of C. jejuni to penetrate the eggs in this study suggests that serious problems may occur under natural field conditions, which may cause significant problems for producers.     

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.     

Fatty acid profile of hair lambs and their crossbreds slaughtered at different weights

This study aims to investigate the effect of breed and slaughter weight on fatty the acid profile of the Longissimus dorsi muscle in lambs. Twenty-four Santa Inês (SI), 24.5 Ile de France × 0.5 Santa Inês (Ile × SI) and 12.5 Texel × 0.5 Santa Inês (Te × SI), slaughtered at different weights (30, 35, 40, and 45 kg), were evaluated. The animals were reared with creep feeding to weaning and were feedlot finished, receiving a diet composed of 30% hay and 70% concentrate. The analyses of fatty acids were carried out on the Longissimus muscle of the 13th rib. The experiment was in a three by four factorial design. The total mean saturated fatty acid level was 44.88%, with 43.30% monounsaturated fats and 1.72% polyunsaturated. The major fatty acids found included oleic (43%), palmitic (22%), and stearic (18%). The concentration of desirable fatty acids varied from 61.56% to 66.78%, with Te × SI (66.78%) having the highest levels (P < 0.05). The slaughter weight affected (P < 0.05) the saturated and unsaturated (both mono and poly) fatty acid profiles. The 35-kg slaughter weight showed the most desirable fatty acid profile. The saturated myristic and stearic fatty acids decreased with an increase in the slaughter weight, but oleic acid increased. In the conditions of the present study, the meat from Santa Ines and Texel × Santa Ines lambs had fatty acid profiles more beneficial for human health due to the higher levels of unsaturated fatty acids.     

Validation of a Leishmania infantum ELISA rapid test for serological diagnosis of Leishmania chagasi in dogs

Canine visceral leishmaniasis (CVL) is caused by Leishmania donovani complex parasites including L. donovani, Leishmania infantum and Leishmania chagasi. As some studies suggest that L. chagasi and L. infantum may be very similar or even the same species, the aim of the present study was to evaluate a commercial rapid ELISA test, originally designed for L. infantum, in the diagnosis of CVL in dogs naturally infected by L. chagasi. A total of 400 serum canine samples, including 283 positive dogs for CVL from an endemic area, 86 clinically healthy dogs from a non-endemic area and 31 dogs seropositive for confounding infectious agents (Trypanosoma cruzi, Toxoplasma gondii, Neospora caninum, Babesia canis and Ehrlichia canis) were used for test validation. An overall sensitivity of 94.7% (95% CI=91.41-97.01%) and specificity of 90.6% (95% CI=83.80-95.21%) was found, with a high degree of agreement (k=0.8445) to the indirect ELISA. When confounding infectious diseases were excluded, specificity increased to 100% (95% CI=95.8-100%), with a higher degree of agreement (k=0.8928). In conclusion, the commercial kit designed for L. infantum was a highly sensitive and specific device for detection of L. chagasi infection in dogs, which indicates high immunoreactivity similarities between L. infantum and L. chagasi.     

Temporal IgG subclasses response in dogs following vaccination against Leishmania with Leishmune®

Canine Visceral Leishmaniasis (CVL) is a widespread zoonotic disease with mandatory euthanasia of infected dogs determined by the Brazilian Ministry of Health. Development of vaccines against CVL may provide a prophylactic barrier, but transitory peak of antibody response detected by standard diagnostic techniques in vaccinated dogs may be interpreted as natural infection. Accordingly, the aim of the present study was to sequentially evaluate total and IgG subclasses response between naturally Leishmania-infected and dogs vaccinated with Leishmune(?). A total of 172 mongrel dogs were divided in four groups: Group 1 (G1) with 45 clinically healthy dogs, Group 2 (G2) and Group 3 (G3) with 45 dogs naturally infected by Leishmania sp. each, symptomatic and asymptomatic respectively, and G4 (G4) with 37 healthy dogs submitted to a complete protocol of a commercially available vaccine against CVL, monitored and evaluated in 5 different chronological moments (M0-M4) up to 180 days after M0. Total IgG, IgG1 and IgG2 were unable to differentiate between infected (G2 and G3) and vaccinated (G4) dogs, demonstrating that polyclonal commercial antibodies do not distinguish these groups apart. Total and IgG subclasses antibodies were not detected until 21 days of the second vaccination dose; however, seroconversion was observed on 21 days and sustained positivity up to 6 months after the vaccination start. A peak of antibodies response was observed on 90 days (M3), when results for total IgG, IgG1, IgG2, IgG3 and IgG4 where highly significant when compared to M0 (P<0.0001). Neither total IgG nor IgG1 effectively differentiated between infected (G2 and G3) and vaccinated (G4) dogs. In conclusion, despite dogs may test serologically negative immediately after vaccination against CVL with Leishmune(?), subsequent seroconversion, antibody peak and positivity up to six months may lead vaccinated dogs to be mistakenly identified as naturally infected dogs during this period.     

Genotypic characterization of Toxoplasma gondii in sheep from Brazilian slaughterhouses: new atypical genotypes and the clonal type II strain identified

Toxoplasma gondii strains are genetically diverse in South America. To date, hundreds of T. gondii isolates from different animal hosts were genotyped in Brazil, most of them are different from those identified around the world. This study aimed to determine T. gondii infection rate in sheep from Brazilian slaughterhouses, as well as the genotype of these isolates. T. gondii antibodies were detected in 66/602 (10.96%) serum samples through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT). MAT-HS and IFAT-IgG presented high concordance (0.95) and strong correlation (r=0.79). T. gondii DNA was detected in tissue samples of 33% (22/66) serum positive sheep by PCR of the 529 bp repetitive element. In the bioassay in mice, T. gondii were detected in mice brain or muscle tissues in 30% (20/66) of serum positive sheep. Positive samples were typed through Restriction Fragment Length Polymorphism (RFLP-PCR) using 11 markers: SAG1, SAG2 (5'-3'SAG2 and alt.SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-6, PK1, Apico and CS3. Of 22 samples, 13 were positive and 9 genotypes were identified. Four of these 9 genotypes are unique. Nine samples had negative results in RFLP-PCR typing, which may be due to low DNA concentration. Six isolates were virulent killing mice between 12 and 25 days postinfection. Two non-virulent isolates belonged to clonal type II genotype, which were not observed in Brazil previously. These findings confirm the high diversity and high frequency of virulent genotypes among Brazilian animals. This study also proved the presence of type II T. gondii in Brazil.     

Synthesis of a combinatorial library of amides and its evaluation against the fall armyworm, Spodoptera frugiperda

The fall armyworm Spodoptera frugiperda is a polyphagous pest that causes important damage in different regions of America and mainly affects corn crops in both tropical and subtropical areas. Currently, control relies on both transgenic plants and/or chemical pesticides. In this work we describe the preparation of an indexed combinatorial library of amides and its toxic effect by contact against S. frugiperda . (E)-1-(1-Piperidinyl)-3-[4-(trifluoromethoxy)phenyl]-2-propen-1-one was the most active compound with an LD(50) = 0.793 μg mg(-1) of larva. This amide was also evaluated by ingestion and at the lowest concentration (1 mg kg(-1)) achieved 83.3% mortality.     

Cross comparison of seminal plasma proteins from cattle and buffalo (Bubalus bubalis)

The objective of this study was to evaluate seminal plasma proteins from cattle and buffalo (Bubalus bubalis), to identify differences between related species. Sixteen buffaloes and 16 cattle between 30 and 60 months of age were used. Semen collection was performed by electroejaculation, followed by macroscopic and microscopic subjective analyses. After analysis, the samples were centrifuged at 800 g for 10 min, and the supernatant (seminal plasma) was recentrifuged at 10,000 g for 30 min at 4°C. The total protein concentration was determined by the Bradford method, and the proteins were digested in solution for mass spectrometry (nLC-MS/MS). Multivariate statistical analysis was used to evaluate the proteomics results by non-hierarchical clustering the considering exponentially modified protein abundance index (emPAI). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used for clustering. Proteomics identified 78 proteins, and multivariate analysis showed 4 that were over-expressed in buffaloes (cystatin C, prosaposin, peptide YY and keratin type II cytoskeletal 5) and 9 in cattle (spermadhesin-1, seminal plasma protein PDC-109, ribonuclease 4, metalloproteinase inhibitor 2, acrosin inhibitor 1, seminal ribonuclease, C-type natriuretic peptide, angiogenin-1 and osteopontin). Among the proteins identified in seminal plasma, the C-type natriuretic peptide and metalloproteinase inhibitors were described for the first time in buffaloes. Some protease inhibitors were found over-expressed in buffaloes, and important proteins in seminal plasma of cattle were not identified or were found at lower expression levels in buffaloes, which can contribute to reproductive performance in this species.