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This study aimed to compare AFM1 occurrence in different cheese types produced by organic and conventional systems; and to evaluate the risk of food exposure to AFM1. A total of 176 commercial cheeses of 17 types were analyzed, 84 of organic and 92 of conventional production. Determination of AFM1 was performed by high- performance liquid chromatography (HPLC), being detected in 30.5% of samples, with 4.8% of organic cheese samples presenting quantifiable AFM1 values between 0.88 and 1.50 µg/kg. On the other hand, 4.3% of conventional cheese samples with values between 0.79 and 6.70 µg/kg. Two conventional cheese samples were above the limit of AFM1 allowed for cheeses by the Brazilian legislation. No statistical difference were found between organic and conventional cheeses regarding the occurrence (p = 0.1780) and concentration of AFM1 (p = 0.1810), according to the Chi-square and the T test, respectively. Estimated daily intake (EDI) and hazard index (HI) of dietary exposure to AFM1 were 0.26 ng/kg/day and 1.28 ng/kg/day, respectively, for conventional cheese samples, and 0.09 ng/kg/day and 0.47 ng/kg/day for organic samples, with no statistical difference for EDI (p = 0.1729) and HI (p = 0.1802) between the two production systems. Comparison between several cheese types from conventional and organic systems indicated that AFM1 is an obstacle to dairy production. Control and prevention of AFM1 contamination, as well as detoxification methods in the final products, are necessary. In the case of organic products, additional research is needed in order to determine which control and detoxification methods should be allowed in this production system.

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Cyprinid herpesvirus 3 has a worldwide distribution and presents high mortality rates in species of Cyprinus carpio, causing serious economic loss to the global aquaculture industry. The description of this infection in other ornamental fish species is still limited. For this purpose, 100 ornamental fish from 24 different species were tested by PCR for Cyprinid hespesvirus 3 and the positive samples represented 6% of the tested samples. Phylogenetic reconstruction, based on the Thymidine Kinase gene, revealed the existence of two distinct clades. One clade grouped a Brazilian sample with European and Asian genotypes of CyHV-3 and a second clade, containing only Brazilian sequences described in this study. All of the Brazilian sequences showed identity values greater than 97.7% when compared to each other. This is the first report of the occurrence of Cyprinid herpesvirus 3 in ornamental fish species in Brazil. These results in association with further studies of viral isolation and characterization can help in establishing effective surveillance and disease control program.

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In recent years, annual cases of gastroenteritis have been reported in the world at high rates, suggesting an association with the consumption of shellfish with enteric viruses in their tissues. Anthropic activities are considered a source of environmental pollution and the main responsible for contamination by pathogenic microorganisms in aquatic environments. The objective of this study was to evaluate, by RT-semi-nested PCR, the presence of astrovirus (AstV) and norovirus genogroup II (NoV GII) in mussels (Mytella falcata) and oysters (Crassostrea brasiliana) collected in two sites of the Lagunar Complex of Cananéia, State of São Paulo, Brazil. A total of 150 samples of mussels and oysters (75 samples each) were analyzed. AstV was not identified in any shellfish sample. NoV GII was detected in 21 samples (14%), 8 mussel samples (38%), and 13 oyster samples (62%). From the 21 positive samples, 16 were analyzed by nucleotide sequencing. The molecular characterization revealed that Brazilian samples were grouped into clades along with other sequences from Brazil, Japan, and Mexico. There was 93.8-100% amino acid sequence similarity among the samples in this study and > 94.9% when compared with the strains isolated from clinical cases in Brazil. The screening of shellfish for the presence of health-significant enteric viruses can help prevent outbreaks among consumers and contribute to the improvement of the estuarine environment.

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The present study analyzes influence of two cleaning and disinfection protocols on frequency of Aspergillus spp. in broiler facilities. We conducted an observational study, applying two cleaning and disinfection protocols before housing 960 one-day-old broilers randomly allocated in 32 boxes with 30 birds. We considered two types of housing as independent experiments, which differed as to the bedding. First experiment consisted of new bedding materials, which we reused in second experiment. We applied two different treatments, Common treatment included sweeping organic matter, humidifying residual material with low pressure water, washing with water and neutral detergent, with posterior rinsing. European treatment was according to the standards of the manufacturer of disinfectants. Procedures included dry and humid organic matter removal, humidification, washing with high pressure water, application of detergent, rinsing, and application of two compound disinfectants: glutaraldehyde (250 g/L) + formaldehyde (185 g/L); p-chloro-m-cresol (210 g/L). To evaluate the presence of Aspergillus spp., we collected samples from surfaces and equipment before and after cleaning and disinfection, at 7, 14, 28, and 42 days of experimental period. Mycological assessment included streaking on Petri dishes containing AFPA (Aspergillus Flavus and Parasiticus Agar, Sigma Aldrich®). Common cleaning and disinfection treatment led to a higher presence of fungus in floors and walls (83% vs. 0% p = 0.0030 and 50% vs. 0% p = 0.0450) in first and second experiments, respectively. As this study demonstrates by using the European procedures, performing a detailed cleaning and disinfection protocol reduces the presence of Aspergillus spp. in broiler facilities.(AU)

O presente estudo analisou a influência de dois protocolos de limpeza e desinfecção na frequência de Aspergillus spp. em instalações de frangos de corte. Realizou-se estudo observacional, aplicando-se dois protocolos de limpeza e desinfecção antes do alojamento de 960 pintainhos de um dia distribuídos aleatoriamente em 32 boxes com 30 aves. Foram realizados 2 experimentos independentes, que diferiam quanto ao material de cama, sendo nova no primeiro e reutilizada no segundo. Realizou-se dois tratamentos: Comum que incluiu varrição da matéria orgânica, umidificação do material residual com água sob baixa pressão, lavagem com detergente neutro e enxágue. O tratamento Europeu seguiu as recomendações do fabricante dos desinfetantes, incluindo remoção de matéria orgânica seca e úmida, umidificação, lavagem com água sob alta pressão, aplicação de detergente, enxágue e aplicação de dois desinfetantes compostos: glutaraldeído (250 g / L) + formaldeído (185 g / L); p-cloro-m-cresol (210 g / L). Foram coletadas amostras de superfícies e equipamentos antes e após a limpeza e desinfecção, aos 7, 14, 28 e 42 dias do período experimental para avaliação micológica, realizada em placas de Petri contendo AFPA (Aspergillus Flavus e Parasiticus Agar, Sigma Aldrich®). O tratamento Comum obteve maior presença de fungos nos pisos e paredes (83% vs. 0% p = 0,0030 e 50% vs. 0% p = 0,0450) no primeiro e segundo experimentos, respectivamente. Este estudo demonstra que a execução de um protocolo detalhado de limpeza e desinfecção, como o tratamento Europeu, reduz a presença de Aspergillus spp. em instalações de frangos de corte.(AU)

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RESUMO

The present study analyzes influence of two cleaning and disinfection protocols on frequency of Aspergillus spp. in broiler facilities. We conducted an observational study, applying two cleaning and disinfection protocols before housing 960 one-day-old broilers randomly allocated in 32 boxes with 30 birds. We considered two types of housing as independent experiments, which differed as to the bedding. First experiment consisted of new bedding materials, which we reused in second experiment. We applied two different treatments, Common treatment included sweeping organic matter, humidifying residual material with low pressure water, washing with water and neutral detergent, with posterior rinsing. European treatment was according to the standards of the manufacturer of disinfectants. Procedures included dry and humid organic matter removal, humidification, washing with high pressure water, application of detergent, rinsing, and application of two compound disinfectants: glutaraldehyde (250 g/L) + formaldehyde (185 g/L); p-chloro-m-cresol (210 g/L). To evaluate the presence of Aspergillus spp., we collected samples from surfaces and equipment before and after cleaning and disinfection, at 7, 14, 28, and 42 days of experimental period. Mycological assessment included streaking on Petri dishes containing AFPA (Aspergillus Flavus and Parasiticus Agar, Sigma Aldrich®). Common cleaning and disinfection treatment led to a higher presence of fungus in floors and walls (83% vs. 0% p = 0.0030 and 50% vs. 0% p = 0.0450) in first and second experiments, respectively. As this study demonstrates by using the European procedures, performing a detailed cleaning and disinfection protocol reduces the presence of Aspergillus spp. in broiler facilities.

O presente estudo analisou a influência de dois protocolos de limpeza e desinfecção na frequência de Aspergillus spp. em instalações de frangos de corte. Realizou-se estudo observacional, aplicando-se dois protocolos de limpeza e desinfecção antes do alojamento de 960 pintainhos de um dia distribuídos aleatoriamente em 32 boxes com 30 aves. Foram realizados 2 experimentos independentes, que diferiam quanto ao material de cama, sendo nova no primeiro e reutilizada no segundo. Realizou-se dois tratamentos: Comum que incluiu varrição da matéria orgânica, umidificação do material residual com água sob baixa pressão, lavagem com detergente neutro e enxágue. O tratamento Europeu seguiu as recomendações do fabricante dos desinfetantes, incluindo remoção de matéria orgânica seca e úmida, umidificação, lavagem com água sob alta pressão, aplicação de detergente, enxágue e aplicação de dois desinfetantes compostos: glutaraldeído (250 g / L) + formaldeído (185 g / L); p-cloro-m-cresol (210 g / L). Foram coletadas amostras de superfícies e equipamentos antes e após a limpeza e desinfecção, aos 7, 14, 28 e 42 dias do período experimental para avaliação micológica, realizada em placas de Petri contendo AFPA (Aspergillus Flavus e Parasiticus Agar, Sigma Aldrich®). O tratamento Comum obteve maior presença de fungos nos pisos e paredes (83% vs. 0% p = 0,0030 e 50% vs. 0% p = 0,0450) no primeiro e segundo experimentos, respectivamente. Este estudo demonstra que a execução de um protocolo detalhado de limpeza e desinfecção, como o tratamento Europeu, reduz a presença de Aspergillus spp. em instalações de frangos de corte.

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The aim of this study is to report the occurrence of Lymphocystivirus in Brazilian ornamental fish. From 25 ornamental fish species submitted for molecular diagnosis, only one sample (Pomacanthus imperator) was positive, and its viral nucleotide sequence obtained clustered with sequences of genotype VII. To our knowledge, this is the first report on the genetic characterization of Lymphocystivirus in Brazil.

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Ranaviruses (family Iridoviridae) cause important diseases in cold-blooded vertebrates. In addition, some occurrences indicate that, in this genus, the same virus can infect animals from different taxonomic groups. A strain isolated from a Ranavirus outbreak (2012) in the state of Sao Paulo, Brazil, had its genome sequenced and presented 99.26% and 36.85% identity with samples of Frog virus 3 (FV3) and Singapore grouper iridovirus (SGIV) ranaviruses, respectively. Eight potential recombination events among the analyzed sample and reference FV3 samples were identified, including a recombination with Bohle iridovirus (BIV) sample from Oceania. The analyzed sample presented several rearrangements compared to FV3 reference samples from North America and European continent. We report for the first time the complete genome of Ranavirus FV3 isolated from South America, these results contribute to a greater knowledge related to evolutionary events of potentially lethal infectious agent for cold-blooded animals.

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Picobirnaviruses (PBVs) are emerging and opportunistic viruses with possible zoonotic potential. In this study, we present the detection, molecular characterization, and genotypic differentiation of PBVs from genogroup I in bovine stool samples from different Brazilian regions. A high proportion of PCR-positive samples (23.4%) was detected in a total of 77 analyzed. Nucleotide identity, alignment, and phylogenetic analyses revealed high diversity among the studied sequences. The results obtained indicate, for the first time, the circulation of bovine PBVs belonging to genogroup I in different Brazilian states, with heterogeneous phylogenetic-clustering profiles.

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Megalocytiviruses have a worldwide distribution, causing serious economic loss to the global aquaculture industry. They also present a threat to ornamental fish trade because megalocytiviral infections have unspecified symptoms, making early diagnosis difficult. In this study, 100 ornamental fish from 24 different species were tested by PCR for megalocytivirus, with a 47% positive rate being identified. Phylogenetic reconstruction, based on the major capsid protein (MCP) gene, clustered all Brazilian samples into a single clade, showing identity values ranging from 99% to 100% when compared to each other. This is the first report of megalocytivirus infection in some ornamental fish species in Brazil.

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It is suggested that Bovine kobuvirus (BKV) is involved in the etiology of gastroenteric diseases especially among calves; however, this association remains unknown. This study evaluated 216 fecal samples from cattle with and without diarrhea symptoms obtained from different regions of Brazil. A 216 bp fragment of the BKV 3D gene was amplified by RT-PCR in 14.4 % (31/216) of the studied samples, and 17 samples were subjected to nucleotide sequencing. All positive samples were obtained from animals aged less than 5 months, and most of animals presented diarrhea (p < 0.05). Phylogenetic analyses showed that the obtained sequences were grouped within the genogroup 2 of BKV forming subclades specific for each Brazilian municipality sampled. In addition, the alignment of the sequences revealed differences of nucleotides between sequences from different locations. Our results indicate for the first time that there is a regional genotypic differentiation of BKV in Brazil.

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Bovine astrovirus (BoAstV) is associated with gastroenterical disorders such as diarrhea, particularly in neonates and immunocompromised animals. Its prevalence is >60 % in the first five weeks of the animal's life. The aim of this study was to detect and perform a phylogenetic analysis of BoAstV in Brazilian cattle. A prevalence of 14.3 % of BoAstV in fecal samples from 272 head of cattle from different Brazilian states was detected, and 11 samples were analyzed by nucleotide sequencing. The majority of positive samples were obtained from diarrheic animals (p < 0.01). Phylogenetic analysis revealed that Brazilian samples were grouped in clades along with other BoAstV isolates. There was 74.3 %-96.5 % amino acid sequence similarity between the samples in this study and >74.8 % when compared with reference samples for enteric BoAstV. Our results indicate, for the first time, the occurrence of BoAstV circulation in cattle from different regions of Brazil, prevalently in diarrheic calves.

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As bactérias psicrotróficas podem afetar a qualidade de diversos produtos lácteos. A relação entre a contagem de psicrotróficos no leite pasteurizado e as frações de caseína do leite longa vida foi avaliada. O processo ultra alta temperatura foi conduzido de acordo com a seguinte sequência: pasteurização a 72-75 ºC por 15 s, pré-aquecimento a 85º C, esterilização a 142-145 ºC por 2 s por injeção direta de vapor ao leite, remoção da água condensada, resfriamento a 70 ºC, homogeneização, resfriamento a 20 ºC, envase asséptico em embalagens cartonadas de 1L e armazenamento à temperatura ambiente por 120 dias. As contagens de psicrotróficos foram determinadas no leite pasteurizado por contagem padrão em placas. As frações de caseína (as1, as2, beta e k) foram avaliadas no leite longa vida nos dias 8 e 120 de armazenamento por cromatografia líquida de alta eficiência. Foi observado aumento na quantidade de as2-caseína e redução da as2-caseína como porcentagem da caseína total ao final do armazenamento (P<0,05). Houve correlação positiva (P<0,05) no dia 120 de armazenamento entre psicrotróficos e a fração k-caseína.

Psycrothrophic bacteria can affect the quality of several dairy products. The relationship between psycrothrophic counts in pasteurized milk and casein fractions of ultra-high-temperature (UHT) milk was evaluated. The UHT process was performed according to the following sequence: pasteurization at 72-75 oC for 15 s, pre-heating at 85 oC, sterilization at 142-145 ºC for 2 s by direct steam injection into milk, removing of the condensed water, cooling to 70 ºC, homogenization, cooling to 20 ºC, bottling aseptically in 1-L sterile carton boxes and storage at room temperature for 120 days. Psycrothrophic counts were determined in pasteurized milk by standard plate count. Casein fractions (as1, as2, beta and k) were analyzed in UHT milk on days 8 and 120 by high performance liquid chromatography. It was observed increase in quantity of as2-casein and reduction of as2-casein as a percentage of total casein at the end of storage period (P<0,05). There was a positive correlation (P<0,05) on day 120 of storage between psychrotrophic and the k-casein fraction (P<0.05).

RESUMO

In the present study, composition, functional properties and sensory characteristics of Mozzarella cheese produced from milk with somatic cell counts (SCC) at low (<200,000 cells/mL), intermediate (≈400,000 cells/mL) and high (>800,000 cells/mL) levels were investigated. Three batches of cheese were produced for each SCC category. The cheeses were vacuum packed in plastic bags and analysed after 2, 9, 16, 23 and 30 days of storage at 4ºC. SCC level did not affect the moisture, fat, total protein and ash content, mesophilic and psychrotrophic bacteria, and sensory parameters of Mozzarella cheese. However, meltability increased in cheese manufactured from high SCC milk. Results indicated that raw milk used to produce Mozzarella cheese should not contain high SCC (>800,000 cells/mL) in order to avoid changes in the functional properties of the Mozzarella cheese.

No presente estudo foram investigadas a composição, as propriedades funcionais e as características sensoriais do queijo Mussarela produzido a partir de leite com contagens de células somáticas (CCS) em níveis baixos (<200.000 CS/mL), intermediários (≈400.000 CS/mL) e altos (>800.000 CS/mL). Foram produzidos 3 lotes de queijo para cada CCS. Os queijos foram embalados a vácuo e analisados após 2, 9, 16, 23 e 30 dias de armazenamento a 4ºC. O nível de CS não afetou a umidade, os teores de gordura, proteína total e cinzas, os níveis de bactérias mesófilas e psicrotróficas, e os parâmetros sensoriais do queijo Mussarela. Entretanto, houve aumento da capacidade de derretimento no queijo fabricado com leite de alta CCS. Os resultados indicam que o leite cru utilizado para a produção de queijo Mussarela não deve conter níveis de CS acima de 800.000/mL, para evitar alterações nas propriedades funcionais do queijo Mussarela.