Showing papers by "Ministry of Agriculture published in 2018"

Combining extraction and purification steps in sample preparation for environmental matrices: A review of matrix solid phase dispersion (MSPD) and pressurized liquid extraction (PLE) applications

Abstract: The search for adequate sample preparation methods is a permanent concern for the analysis of contaminants and residues analysis in environment and biological samples. Ideally, a sample preparation method must be capable to perform extraction and purification in a simultaneous way. Although several methodologies have been developed in recent years, meeting the above proposed criteria, the overall aim of our review is to discuss two sample preparation techniques and their recent trends: matrix solid-phase dispersion (MSPD) and pressurized liquid extraction (PLE), focusing in the simultaneous extraction and clean-up steps.

Determination of pharmaceutical compounds in hospital wastewater and their elimination by advanced oxidation processes.

Abstract: This study investigates the mineralization efficiency, i.e. removal of total organic carbon (TOC) in hospital wastewater by direct ozonation, ozonation with UV radiation (O3/UV), homogeneous cataly...

Phenolic Compounds Determined by LC-MS/MS and In Vitro Antioxidant Capacity of Brazilian Fruits in Two Edible Ripening Stages.

Abstract: The aim of this study was to investigate the free individual phenolics and the in vitro antioxidant capacity of blackberry, acerola, yellow guava, guabiju, jambolan and jabuticaba fruits in two edible stages. Of the thirty-three phenolics investigated by liquid chromatography - tandem mass spectrometry (LC-MS/MS), twenty-five were quantified and the major ones were catechin, isoquercitrin, epicatechin and gallic acid. The highest values for the total phenolic content (in dry matter) were observed for acerola (83.6 to 97.7 mg gallic acid equivalents g−1 DM) and blackberry (18.9 to 28.3 mg gallic acid equivalents g−1 DM); however, acerola, jabuticaba, and blackberry showed the highest antioxidant capacities (134.6 to 1120.4 mg Trolox equivalents g−1 for 2,2-diphenyl-1-picrylhydrazyl and 43.6 to 501.8 μmol Trolox equivalents g−1 for ferric reducing antioxidant power). For most fruits, the antioxidant capacity decreased during the ripening, possibly due to a decrease in the concentration of most of the phenolics.

HoBi-like is the most prevalent ruminant pestivirus in Northeastern Brazil

Abstract: The ruminant pestiviral species BVDV-1, BVDV-2 and BDV, along with the putative species HoBi-like, may cause substantial economic losses in cattle, sheep and goats. Brazil's large size, variable biomes and wide range of ruminant animal production within different geographic regions suggest that the presence and prevalence of ruminant pestivirus may differ by regions within Brazil. This study investigated the genetic diversity of ruminant pestiviruses and determined the frequency of active infections within two states of the Northeast Region of Brazil, Maranhao and Rio Grande do Norte. Serum samples from 16,621 cattle and 2,672 small ruminants from 569 different herds residing in this region were tested by RT-PCR followed by DNA sequencing. Seventeen positive cattle were detected (0.1%) from fifteen different herds (2.64%). All isolates were classified as HoBi-like pestiviruses based on phylogenetic analysis. All small ruminant samples tested negative. The findings presented herein suggest that the Northeast Region of Brazil has a uniquely high prevalence of HoBi-like viruses. The increasing reports of HoBi-like viruses detected in cattle in the field suggest that natural infection with these viruses may be more widespread than previously thought. The identification of HoBi-like viruses as the most prevalent type of ruminant pestivirus circulating in the Northeast Region of Brazil indicates the need for both continued monitoring and determination of the extent of economic losses associated with HoBi-like virus infections. In addition, it must be taken into account in the choice of diagnostic tests and in vaccine formulations.

Wide-Scope Determination of Pharmaceuticals and Pesticides in Water Samples: Qualitative and Confirmatory Screening Method Using LC-qTOF-MS

Abstract: Aquatic system contamination is a subject of concern due to the high number of contaminants with the potential to be hazardous for plant and animal species, including humans. A large number of analytical methods have been developed to evaluate the extension of water resource pollution, most of them focused on target residue analysis. Screening methods for potential target and non-target compounds are a potential alternative and could be used for a comprehensive evaluation of samples composition. Liquid chromatography coupled to time of flight mass spectrometry (LC-qTOF-MS/MS) was developed to provide full-spectrum data and high mass resolution with appropriate selectivity and sensitivity for monitoring of 300 pharmaceuticals and pesticides in environmental water samples. The validation was based on anti-doping analysis validation protocols, using three different concentration levels (0.01, 0.1, and 1.0 μg L−1). Confirmation analysis was achieved using two fragment ions monitoring for each analyte. Samples were extracted by solid-phase extraction and analyzed by LC-qTOF-MS/MS. The final method was able to detect 170 chemicals in wastewater treatment plant effluent and 198 chemicals in surface water. The advantage of using post-acquisition data processing provided by a qTOF-MS/MS system allowed for non-target analysis for an illicit drug metabolite determination. The method was successfully applied to real samples of wastewater and surface water.

Outbreaks, toxicology, and analytical methods of marine toxins in seafood

Abstract: The consumption of seafood with marine toxins can cause intoxications of varied severity and often-fatal symptoms which may appear in few minutes. Marine toxins are resistant to cooking and freezing, posing a serious hazard to public health. They are usually classified as paralytic, neurotoxic, amnesic, or diarrheic shellfish poisoning. Azaspiracids are a separate group whose poisoning symptoms resemble the diarrheic group. Moreover, some fish species may be contaminated with tetrodotoxin and ciguatoxins. Seafood and seawater control is practiced to protect the consumers’ health and to avoid damages to the fishing economy, due to closures of cultivated areas. Some of the main outbreaks and toxicological properties of marine toxins, as well as the main analytical control tools available were reviewed.

Risk assessment of 22 chemical elements in dry and canned pet foods

Abstract: There is little information on the levels of chemical elements in pet food considering the dietary requirements as well as risk assessment of toxicity. This study aimed to determine the essential and toxic elements in dry and canned foods for dogs and cats and estimate their daily intake. We compared the levels of the chemical elements between the dry and wet (canned) food to the levels recommended by the Association of American Feed Control Officials (AAFCO) and the European Pet Food Industry Federation (FEDIAF) and the maximum tolerable level proposed by European Commission (EC). In addition, the estimated daily intake (EDI) for each one of the elements through food was calculated. Seventy-six dry food samples (dogs n = 62 and cats n = 14) from 43 brands and 12 canned foods (dogs n = 6 and cats n = 6) from 5 brands, were purchased from Brazilian supermarkets. Mean levels of all essential elements reached the minimum level recommended by AACFO. Selenium levels were very close to the maximum limit proposed by AAFCO. Besides, the iron concentrations in canned (moist) food were statistically higher than in dry food and its EDI for cats (54 mg/day × kg body weight) exceeded the maximum limit recommended by FEDIAF. Regarding the toxic metals, the concentrations of mercury and cadmium, in dry and canned food, were considerably higher than the maximum tolerable level proposed by EC. Overall, the results show that levels of essential elements are in agreement with the nutrient requirement. On the other hand, mercury and cadmium in pet food are an issue of concern.

Risk factors for human Mycobacterium bovis infections in an urban area of Brazil

Abstract: BACKGROUND The World Health Organization (WHO) has classified human zoonotic tuberculosis (TB) due to Mycobacterium bovis as a neglected issue in the developing world. In a recent cross-sectional study in Brazil, three of 189 TB patients presented with a coinfection of M. bovis and M. tuberculosis and were selected as cases for this study. OBJECTIVE The aim was to evaluate risk factors (RF) for zoonotic TB in an urban area of Brazil in order to guide preventive programmes. METHODS A matched case-control study was carried out nested within a cross-sectional study. For each of the three cases, 14 age- and sex-matched controls (TB due to M. tuberculosis) were selected. FINDINGS Zoonotic potential exposures (ZE) and extrapulmonary TB (EPTB) were independently associated with zoonotic TB in multivariate analyses. CONCLUSIONS ZE by occupation and consumption of raw milk and derivative products that place individuals in direct and indirect contact with animals and their excretions/secretions increase the risk for zoonotic TB in Brazil, especially among those with EPTB. Therefore, measures such as efficient control of bovine TB, distribution of pasteurised milk and its derivative products, and the diagnosis and monitoring of zoonotic TB in humans are essential steps, especially in developing countries where bovine TB is enzootic, and further studies are necessary.

Liquid Chromatography-Tandem Mass Spectrometry Determination and Depletion Profile of Chlortetracycline, Doxycycline, and Oxytetracycline in Broiler Chicken Muscle After Oral Administration

Abstract: A simple and fast method based on liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) was developed and validated for determination of tetracyclines in broiler chicken muscle. Sample preparation was performed using extraction with acetonitrile, followed by low-temperature purification (at − 20 °C) and further concentration. The chromatographic analysis was carried out using a Zorbax SB-C18 column with gradient elution using water and methanol both acidified with 0.025 M of formic acid. Mass spectrometry with electrospray ionization was operated in positive polarity using selected reaction monitoring (SRM) analysis mode, achieving the requirements of four identification points for each compound. Demeclocycline was used as internal standard. The method validation was done according to the criteria of Commission Decision 2002/657/EC. Parameters such as recovery, matrix effects, selectivity/specificity, linearity, precision (intra- and inter-day precision), accuracy, limits of detection (LOD) and quantification (LOQ), decision limit (CCα), detection capability (CCβ), and robustness were determined. Intra-day precision values were within the range 2.2–5.8% and inter-day precision was less than 10% for all analytes. Accuracy ranged from 98.2 to 103.2%. The method was successfully applied for depletion studies of chlortetracycline, doxycycline, and oxytetracycline in broiler chicken tissues after multiple oral administrations. After the depletion studies, the present study support more prudent use of CTC, DOX, and OTC for treatment of chickens and suggest a dose of 60 mg kg−1 body weight for CTC and OTC and 20 mg kg−1 body weight for DOX, orally administrated for five consecutive days.

Comprehensive evolutionary and phylogenetic analysis of Hepacivirus N (HNV).

Abstract: Hepaciviruses (HVs) have been detected in several domestic and wild animals and present high genetic diversity. The actual classification divides the genus Hepacivirus into 14 species (A–N), according to their phylogenetic relationships, including the bovine hepacivirus [Hepacivirus N (HNV)]. In this study, we confirmed HNV circulation in Brazil and sequenced the whole genome of two strains. Based on the current classification of HCV, which is divided into genotypes and subtypes, we analysed all available bovine hepacivirus sequences in the GenBank database and proposed an HNV classification. All of the sequences were grouped into a single genotype, putatively named ‘genotype 1’. This genotype can be clearly divided into four subtypes: A and D containing sequences from Germany and Brazil, respectively, and B and C containing Ghanaian sequences. In addition, the NS3-coding region was used to estimate the time to the most recent common ancestor (TMRCA) of each subtype, using a Bayesian approach and a relaxed molecular clock model. The analyses indicated a common origin of the virus circulating in Germany and Brazil. Ghanaian sequences seemed to have an older TMRCA, indicating a long time of circulation of these viruses in the African continent.

First report of Ornithodoros peropteryx in Brazil, and the occurrence of Ornithodoros cavernicolous in the western Brazilian Amazon

Abstract: More than half of the Brazilian soft ticks (Argasidae) are represented by species associated with bats. Based on a morphological and molecular approach of free-living ticks collected inside two unexplored caves in Rondonia state, western Brazilian Amazon, we report the occurrence of Ornithodoros peropteryx for the first time in Brazil. Our findings also extend the distribution of Ornithodoros cavernicolous towards western Brazilian Amazon. Additionally, our collections include previously unknown male of O. peropteryx. In both species, partial 16S rRNA mitochondrial gene sequences showed >99% of identity when compared to available conspecific data from other localities. Although more data is necessary to make a general conclusion, high vagility of hosts could be favoring low genetic structure among these ectoparasites.

First record of the soybean stem fly Melanagromyza sojae (Diptera: Agromyzidae) in the Brazilian Savannah

Abstract: The soybean stem fly [ Melanagromyza sojae (Diptera: Agromyzidae)] is a pest widely distributed in the Asian continent. In Brazil, its occurrence has been reported since the 1980s, but it had not been reported in Brazilian Savannah areas yet. This is the first record of M. sojae in this region of great importance for the soybean world production. Specimens of the soybean stem fly were collected in a volunteer soybean area, in April 2018, in the Goias state, Brazil, and sent for molecular identification. The similarity index of the collected specimens with M. sojae was higher than 99 %, thus confirming the identification of the species. The infestation level in the collection area was approximately 100 % of plants with damage caused by this insect. In Brazil, the occurrence of M. sojae , up to the present time, had been described as restricted to the southern states. This record raises the hypothesis that M. sojae is widespread in other agricultural areas of the Brazilian Savannah, and that its dissemination is favored by the maintenance of volunteer soybean.

Prevalência de leptospirose em rebanhos bovinos no Pantanal de Mato Grosso do Sul

Abstract: RESUMO: Foi realizado um estudo epidemiologico da leptospirose em femeas acima de 24 meses, provenientes de 246 rebanhos, e 2.766 animais amostrados aleatoriamente nos nove municipios que compoem a regiao do Pantanal de Mato Grosso do Sul, bem como identificados os fatores de risco associados a doenca. As amostras de sangue foram coletadas no periodo de setembro a novembro de 2009 e examinadas pelo teste de aglutinacao microscopica ante uma colecao de 24 antigenos vivos de Leptospira spp., representantes dos sorovares Australis, Bratislava, Autumnalis, Butembo, Castellonis, Batavie, Canicola, Whitcombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Javanica, Panama, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi, Sentot, Andamana e Patoc. Adicionalmente, representantes de doze estirpes de leptospiras isoladas no Brasil foram adicionados a colecao de antigenos do teste de soroaglutinacao microscopica (SAM). A prevalencia aparente foi de 66% e a prevalencia real de animais infectados, de 79,80%, com intervalo de confianca (IC) de 95% (78,3-81,3) e 241 rebanhos apresentando pelo menos um animal reagente. Os sorovares mais provaveis foram o Hardjo seguido pelo Wolffi. Os resultados demonstram que a leptospirose bovina continua presente no Pantanal, com alta prevalencia tanto em rebanhos quanto em individuos, sendo os principais fatores de risco para a doenca o tipo de exploracao e a raca.

Virome analyses of Hevea brasiliensis using small RNA deep sequencing and PCR techniques reveal the presence of a potential new virus.

Abstract: Hevea brasiliensis is an important commercial crop due to the high quality of the latex it produces; however, little is known about viral infections in this plant. The only virus described to infect H. brasiliensis until now is a Carlavirus, which was described more than 30 years ago. Virus-derived small interfering RNA (vsiRNAs) are the product of the plant’s antiviral defense triggered by dsRNA viral intermediates generated, during the replication cycle. These vsiRNAs are complementar to viral genomes and have been widely used to identify and characterize viruses in plants. In the present study, we investigated the virome of leaf and sapwood samples from native H. brasiliensis trees collected in two geographic areas in the Brazilian Amazon. Small RNA (sRNA) deep sequencing and bioinformatic tools were used to assembly, identify and characterize viral contigs. Subsequently, PCR amplification techniques were performed to experimentally verify the presence of the viral sequences. Finally, the phylogenetic relationship of the putative new virus with related viral genomes was analyzed. Our strategy allowed the identification of 32 contigs with high similarity to viral reference genomes, from which 23 exhibited homology to viruses of the Tymoviridae family. The reads showed a predominant size distribution at 21 nt derived from both strands, which was consistent with the vsiRNAs profile. The presence and genome position of the viral contigs were experimentally confirmed using droplet digital PCR amplifications. A 1913 aa long fragment was obtained and used to infer the phylogenetic relationship of the putative new virus, which indicated that it is taxonomically related to the Grapevine fleck virus, genus Maculavirus. The putative new virus was named Hevea brasiliensis virus (HBrV) in reference to its host. The methodological strategy applied here proved to be efficient in detecting and confirming the presence of new viral sequences on a ‘very difficult to manage’ sample. This is the second time that viral sequences, that could be ascribed as a putative novel virus, associated to the rubber tree has been identified.

Prostaglandin A1 triggers Mayaro virus inhibition and heat shock protein 70 expression in an epithelial cell model

Abstract: INTRODUCTION: The Mayaro virus (MAYV), which is an arbovirus closely related to the Chikungunya virus, causes a dengue-like acute illness that is endemic to Central and South America. We investigated the anti-MAYV activity of prostaglandin A1 (PGA1), a hormone which exhibits antiviral activity against both ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) viruses. Further, we examined the effects of inducting the stress protein HSP70 following PGA1 treatment. METHODS: Hep-2 cells infected with MAYV were treated with PGA1 (0.1-6μg/ml) 12h before infection and for different periods post-infection. Inhibition of viral replication inhibition was analyzed via viral titer determination, whereas the effect of PGA1 on viral morphogenesis was examined via transmission electron microscopy (TEM). Autoradiography (with 35S methionine labeling) and western blotting were used to assess the effect of PGA1 treatment on viral and cellular protein synthesis, and on HSP70 induction, respectively. RESULTS: PGA1 strongly reduced viral replication in Hep-2 cells, particularly when added during the early stages of viral replication. Although PGA1 treatment inhibited viral replication by 95% at 24 hours post-infection (hpi), viral structural protein synthesis was inhibited only by 15%. TEM analysis suggested that PGA1 inhibited replication before viral morphogenesis. Western blot and densitometry analyses showed that PGA1 treatment increased HSP70 protein levels, although this was not detectable via autoradiography. CONCLUSIONS: PGA1 inhibits MAYV replication in Hep-2 cells at early stages of viral replication, prior to production of viral structural proteins, possibly via HSP70 induction.