Essential oils isolated from popular medicinal plants and spices as alternative antimicrobial and antibiofilm compounds against the pig pathogen Actinobacilluspleuropneumoniae / Óleos essenciais isolados de plantas medicinais e temperos populares como compostos antimicrobianos e antibiofilme alternativos contra o patógeno de suínosActinobacillus pleuropneumoniae

ABSTRACT: Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

RESUMO: Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

Essential oils isolated from popular medicinal plants and spices as alternative antimicrobial and antibiofilm compounds against the pig pathogen Actinobacilluspleuropneumoniae / Óleos essenciais isolados de plantas medicinais e temperos populares como compostos antimicrobianos e antibiofilme alternativos contra o patógeno de suínosActinobacillus pleuropneumoniae

Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

Presence of antibiotic-resistant bacteria in pigs for consumption: one health challenge: literature review / Presença de bactérias resistentes a antibióticos em suínos para consumo: desafio à saúde única: revisão de literatura

Pig farming is an area of livestock that has been developing the most in Brazil and the world, with production increasing every year, generating jobs, and being of great importance for the Brazilian economy. In swine production, great health enables these animals to reach their highest point of development and antimicrobials are used, either prophylactically or through food, as growth promoters. Within swine culture, there is a concern regarding antibiotic-resistant bacteria; however, Staphylococcus spp. do not receive the necessary prominence in research, since the pathologies caused by them do not tend to cause great economic losses. Therefore, this review aimed to highlight the importance of bacterial resistance within breeding stock, its possible origins, the importance of Staphylococcus spp. within this topic, and its evolution in swine farming over the years. For this, studies were selected, with an emphasis on information such as country, number of samples, presence of Staphylococcus spp. resistant to methicillin, breeding phase, and phenotypic and molecular tests. In addition, publications were selected that show the importance of understanding the biological and resistance profiles of Staphylococcus spp. in swine herds in Brazil and around the world.

A suinocultura é uma das áreas da pecuária que mais se desenvolveu e se desenvolve no Brasil e no mundo, com a sua produção aumentando todos os anos, gerando empregos diretos e indiretos, e sendo de grande importância para a economia brasileira. Destaca-se na produção de suínos a necessidade de grande sanidade para que estes animais alcancem seu maior ponto de desenvolvimento e, para tal, são utilizados antimicrobianos, seja de forma profilática ou através da alimentação, como promotores de crescimento. Dentro da suinocultura também está presente a preocupação envolvendo as bactérias resistentes a antibióticos, contudo, os Staphylococcus spp. não recebem o destaque necessário em pesquisas, já que as patologias causadas por estes ainda não tendem a causar grandes prejuízos econômicos de forma direta aos produtores. Por este motivo, esta revisão objetivou evidenciar a importância da resistência bacteriana dentro dos planteis, suas possíveis origens, a importância dos Staphylococcus spp. dentro deste tópico e a sua evolução na suinocultura através dos anos. Para este foram selecionados trabalhos encontrados em diferentes bases de dados com ênfase em informações como país, número de amostras, presença de Staphylococcus spp. resistentes a meticilina, fase de criação, e testes fenotípicos e moleculares, além de publicações que evidenciam a importância de se conhecer os perfis biológicos e de resistência dos Staphylococcus spp. nos planteis de suínos do Brasil e do mundo.

Early puberty in short-haired Guinea pigs kept in laboratory animal facilities

Lab animals, such as Guinea pigs (Cavia porcellus), are crucial for scientific development, as they play an important role in the development and quality control chain of vaccines and drugs distributed by the Brazilian public health system. Investigating their biological and physiological parameters is fundamental to raise and keep these animals, so the handling of the facilities that hold them can be updated whenever new information comes up, with the well-being of the animals and alignment with the 3 Rs in mind. In the search for understanding reproductive aspects of Guinea pigs, the present study had the main goal of studying puberty by means of estrous cycle analysis in short-haired Guinea pigs. Guinea pigs have a vaginal occlusive membrane that covers the vaginal orifice. Its rupture takes place gradually and naturally, moments before labor and during estrus. The present study followed 42 females as for the presentation of the vaginal occlusive membrane. Once the membranes ruptured spontaneously, a swab was collected to study vaginal cytology. Membrane rupture was observed in 39 females; six females showed membrane rupture with less than 21 days of age (17 to 21 days). Twenty-three females were characterized as being in estrus due to cytology showing a prevalence of anucleated superficial cells. One of these females was younger than 21 days old. The opening of the vaginal occlusive membrane took place most frequently in intervals between 17 and 18 days, and the membrane remained open between one and three consecutive days. It was possible to follow three cycles of membrane opening on six females. The present study showed the need to adapt handling guidelines for C. porcellus kept in research animal facilities. The early age of puberty imposes the need of separate the female daughters from their fathers at 16 days old.(AU)

Diversity of Diptera species associated with pig carcasses in a Brazilian city exposed to high rates of homicide

Abstract Cities in northeastern Brazil experience extreme rates of unsolved homicides, a situation that stimulates innovative procedures in the police work, such as forensic entomology. We surveyed necrophagous insects associated with carrion in a city exposed to high rates of homicides in Northeastern Brazil. The experiments were carried out in a rainforest fragment located in Recife, State of Pernambuco. Two pig carcasses were used as models, one in the dry and the other in the rainy season. The collection of adults was performed daily until the complete skeletonization of the carcasses. At least 32 Diptera species from the families Calliphoridae, Muscidae, Sarcophagidae, Fanniidae, Phoridae, Anthomyiidae, Piophilidae, and Stratiomyidae were registered, some of which have been previously documented on cadavers. A high richness of Diptera species was registered in all stages of decomposition. A strong overlap in the occurrence of most species was observed, which invalidates a defined entomological succession on the carcasses. Two species stood out in terms of abundance: Ophyra chalcogaster (Muscidae) and Chrysomya albiceps (Calliphoridae). The ubiquity of Hemilucilia semidiaphana (Calliphoridae) seems to confirm its preference for forest fragments exposed to low anthropogenic action. Our data contribute to expand the knowledge on the geographical distribution of forensically relevant species in the region and confirm the rapid dissemination of invasive Chrysomya species in forested areas.

Viability of Brazilian distillers dried grains with solubles for pigs / Viabilidade de grãos secos de destilaria com solúveis produzidos no Brasil para suínos

The purpose of this study was to evaluate the performance, carcass, meat and economic viability characteristics of pigs fed Brazilian coproducts of corn ethanol production. A total of 40 finishing pigs were subjected to four levels of distillers dried grains with solubles (DDGS; 0, 100, 200, or 300 g kg-1) for 28 days, and body weight, daily feed intake, daily weight gain and feed conversion were measured. After slaughter, the carcass yield, fat thickness, loin eye area, the depth and perimeter of the loin eye, and the pH and temperature at 45 min and 24 h post-mortem, along with the drip loss and color of the Longissimus lumborum were evaluated. The economic viability of the diets was calculated. The inclusion of DDGS showed a quadratic effect on daily feed intake over 0-14 days. Pigs fed with 300 g kg-1 of DDGS showed worse feed conversion than those fed the control diet over 0-7, 0-14 and 0-21 days. The a* color parameter of the meat decreased linearly with increasing DDGS concentration but the other carcass and meat characteristics were not affected. Inclusion of 184.1 g kg-1 of DDGS showed the best economic viability, considering the cost of weight gain. DDGS may be included in pig diets up to 200 g kg-1 without affecting the performance or carcass and meat characteristics, while 300 g kg-1 compromises feed conversion.(AU)

O propósito deste estudo foi avaliar o desempenho, características de carcaça e carne e viabilidade econômica de suínos alimentados com coprodutos da produção de etanol de milho produzido no Brasil. Um total de 40 suínos em terminação foi distribuído em quatro níveis de DDGS de milho (0, 100, 200, ou 300 g kg-1) por 28 dias e seu peso corporal, consumo de ração diário, ganho de peso diário e conversão alimentar foram mensurados. Após o abate, rendimento de carcaça, espessura de toucinho, área de olho de lombo, profundidade e perímetro do olho de lombo, pH e temperatura a 45 min e 24 h post-mortem além da perda por gotejamento e do Longissimus lumborum foram avaliados. A viabilidade econômica das dietas foi calculada. A inclusão de DDGS mostrou efeito quadrático no consumo de ração em 0-14 dias. Suínos alimentados com 300 g kg-1 de DDGS apresentaram pior conversão alimentar em comparação àqueles da dieta controle em 0-7, 0-14 e 0-21 dias. A cor a* da carne diminuiu linearmente com aumento da concentração de DDGS nas dietas mas as outras características de carcaça e carne não foram afetadas. Inclusão de 184.1 g kg-1 de DDGS mostrou melhor viabilidade econômica, considerando o custo por ganho de peso. Os DDGS podem ser incluídos em dietas de suínos até 200 g kg-1 sem afetar o desempenho e características de carcaça e carne enquanto 300 g kg-1 compromete a conversão alimentar.(AU)

Evaluation of traditional Brazilian sausage (linguiça calabresa) elaborated with oregano and basil extracts as natural antioxidants / Avaliação da linguiça calabresa elaborada com extratos de orégano e manjericão como antioxidantes naturais

This study aimed to evaluate traditional Brazilian sausage (linguiça calabresa) elaborated with oregano and basil extracts as natural antioxidants. Hydroalcoholic extracts of dried oregano and basil leaves were prepared, freeze-dried, and evaluated for antioxidant activity by Folin-Ciocalteu, DPPH, FRAP, and ABTS assays. Freeze-dried oregano extract showed higher (p < 0.05) antioxidant activity than freeze-dried basil extract by all methods. Extracts were used in preparations of seven sausage formulations, as follows: F1, 0.05% oregano extract and 0.05% basil extract; F2, 0.1% oregano extract and 0.05% basil extract; F3, 0.05% oregano extract and 0.1% basil extract; F4, 0.075% oregano extract and 0.075% basil extract, F5, 0.1% oregano extract and 0.1% basil extract; F6 (control), without extract; and F7, 0.01% butylated hydroxytoluene (BHT). Sausage formulations were analyzed for chemical composition, pH, acidity, water activity, lipid oxidation, color, and texture. Formulations showed differences in pH at different storage periods. F1, F2, F3, and F7 had the lowest (p < 0.05) lipid oxidation values at 53 days of storage. F1, F2, F6, and F7 showed higher a* values and lower b* values. Hardness and chewiness decreased (p < 0.05) during storage for all formulations. F1, F2, and F3 had similar efficiency to that with synthetic antioxidant BHT (F7), therefore, oregano and basil extracts have potential application as natural antioxidants in the food industry.(AU)

Este estudo objetivou avaliar linguiça calabresa elaborada com extratos de orégano e manjericão como antioxidantes naturais. Extratos hidroalcoólicos de orégano e manjericão desidratados foram preparados, liofilizados e submetidos a avaliação de sua atividade antioxidante pelos métodos de FolinCiocalteu, DPPH, FRAP e ABTS. O extrato liofilizado de orégano apresentou maior atividade antioxidante (p < 0,05) que o extrato de manjericão para todos os métodos. Os extratos liofilizados foram usados em sete formulações de linguiça calabresa, como segue: F1 com 0,05% de cada extrato, F2 com 0,1% de extrato de orégano e 0,05% de extrato de manjericão, F3 com 0,05% de extrato de orégano e 0,1% de extrato de manjericão, F4 com 0,075% de cada extrato, F5 com 0,1% de cada extrato, F6 (controle), sem adição de extratos e F7 adição de 0,01% de hidroxitolueno butilado (BHT). As linguiças calabresas foram avaliadas, quanto a composição química, pH, acidez, atividade de água, oxidação lipídica, cor e textura. As formulações apresentaram variação de pH nos diferentes períodos de estocagem. As formulações F1, F2, F3 e F7 apresentaram os menores valores de oxidação lipídica (p < 0,05) aos 53 dias de armazenamento. As formulações F1, F2, F6 e F7 apresentaram maiores valores de a* e menores valores de b*. Observouse redução (p < 0,05) de dureza e mastigabilidade ao longo do período de armazenamento para todas as formulações. As formulações F1, F2 e F3 apresentaram eficiência similar à formulação com antioxidante sintético BHT (F7), portanto, os extratos de orégano e manjericão tem potencial como antioxidante natural na indústria de alimentos.(AU)

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production

Abstract The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.

Detection of sapovirus in brazilian pig farms / Detecção de sapovírus em criações de suínos brasileiras

Sapoviruses (Caliciviridae) are considered important agents of gastroenteritis worldwide affecting animals and humans. In pig farming, the epidemiology is not completely understood because it can affect all stages of production, with symptomatic (diarrhea) or asymptomatic pigs. The aim of our study was to investigate Sapovirus occurrence in Brazilian pig farms. A total of 166 fecal samples of pigs, with different ages, from Minas Gerais, São Paulo, and Mato Grosso States were submitted to RT-PCR reactions and confirmed with nucleotide sequencing of Sapovirus RdRp gene. Six (3.61%) samples were positive and four had partial RdRp gene sequenced, putatively belonging to GVII.1 genogroup, also reported in swine herds in Brazil.

Sapovírus (Caliciviridae) são considerados importantes agentes causadores de gastroenterites em todo o mundo, afetando animais e humanos. Na suinocultura, sua epidemiologia ainda não foi totalmente esclarecida, pois pode afetar todas as fases da produção, com suínos sintomáticos (diarreia) ou assintomáticos. O objetivo do nosso estudo foi investigar a ocorrência de Sapovírus em granjas de suínos brasileiras. Um total de 166 amostras fecais de suínos, com diferentes idades, dos estados de Minas Gerais, São Paulo e Mato Grosso foram submetidas a reações de RT-PCR e confirmadas com sequenciamento de nucleotídeos do gene RdRp do Sapovírus. Seis (3,61%) amostras foram positivas e quatro delas tinham sequenciamento parcial do gene RdRp, supostamente pertencente ao genogrupo GVII.1, previamente relatado em rebanhos suínos no Brasil.

Detection of sapovirus in brazilian pig farms / Detecção de sapovírus em criações de suínos brasileiras

Sapoviruses (Caliciviridae) are considered important agents of gastroenteritis worldwide affecting animals and humans. In pig farming, the epidemiology is not completely understood because it can affect all stages of production, with symptomatic (diarrhea) or asymptomatic pigs. The aim of our study was to investigate Sapovirus occurrence in Brazilian pig farms. A total of 166 fecal samples of pigs, with different ages, from Minas Gerais, São Paulo, and Mato Grosso States were submitted to RT-PCR reactions and confirmed with nucleotide sequencing of Sapovirus RdRp gene. Six (3.61%) samples were positive and four had partial RdRp gene sequenced, putatively belonging to GVII.1 genogroup, also reported in swine herds in Brazil.(AU)

Sapovírus (Caliciviridae) são considerados importantes agentes causadores de gastroenterites em todo o mundo, afetando animais e humanos. Na suinocultura, sua epidemiologia ainda não foi totalmente esclarecida, pois pode afetar todas as fases da produção, com suínos sintomáticos (diarreia) ou assintomáticos. O objetivo do nosso estudo foi investigar a ocorrência de Sapovírus em granjas de suínos brasileiras. Um total de 166 amostras fecais de suínos, com diferentes idades, dos estados de Minas Gerais, São Paulo e Mato Grosso foram submetidas a reações de RT-PCR e confirmadas com sequenciamento de nucleotídeos do gene RdRp do Sapovírus. Seis (3,61%) amostras foram positivas e quatro delas tinham sequenciamento parcial do gene RdRp, supostamente pertencente ao genogrupo GVII.1, previamente relatado em rebanhos suínos no Brasil.(AU)

Phosphorus accumulation in a southern Brazilian Ultisol amended with pig manure for nine years

This study evaluated P pools after nine years of successive application of either pig slurry (PS) or deep pig litter (DL) in a no-till Ultisol from southern Brazil. The experiment was established in Dec 2002 with the treatments control, application of 90 and 180 kg N ha−1 N as PS and as DL. In Mar 2010, soil samples were taken at six layers up to 30 cm deep. Total organic and inorganic P were assessed by the ignition method, and P compounds classes were evaluated by 31P-NMR spectroscopy. Total soil P increased proportionally with the P amount applied via DL and PS. Only DL application increased soil organic P, mainly at the highest dose and in the uppermost soil layers. The application of high doses of manure to these soils under no-till to meet crop N demands significantly increased P accumulation at the soil surface, especially with DL. This, in turn, increases the risk of contamination of water bodies due to P transfer from soil to rivers via runoff. The ignition method overestimates organic P compared to P-NMR. The highest proportion of organic P estimated by the ignition and P-NMR methods, at surface layers in the control suggests that inorganic P is added to the plots treated, increasing inorganic P and decreasing organic P. Moreover, with no P additions to the control, inorganic soil P is removed by plants, causing an apparent increase in the organic P proportion.

Phosphorus accumulation in a southern Brazilian Ultisol amended with pig manure for nine years

This study evaluated P pools after nine years of successive application of either pig slurry (PS) or deep pig litter (DL) in a no-till Ultisol from southern Brazil. The experiment was established in Dec 2002 with the treatments control, application of 90 and 180 kg N ha−1 N as PS and as DL. In Mar 2010, soil samples were taken at six layers up to 30 cm deep. Total organic and inorganic P were assessed by the ignition method, and P compounds classes were evaluated by 31P-NMR spectroscopy. Total soil P increased proportionally with the P amount applied via DL and PS. Only DL application increased soil organic P, mainly at the highest dose and in the uppermost soil layers. The application of high doses of manure to these soils under no-till to meet crop N demands significantly increased P accumulation at the soil surface, especially with DL. This, in turn, increases the risk of contamination of water bodies due to P transfer from soil to rivers via runoff. The ignition method overestimates organic P compared to P-NMR. The highest proportion of organic P estimated by the ignition and P-NMR methods, at surface layers in the control suggests that inorganic P is added to the plots treated, increasing inorganic P and decreasing organic P. Moreover, with no P additions to the control, inorganic soil P is removed by plants, causing an apparent increase in the organic P proportion.(AU)

Diversity of Diptera species associated with pig carcasses in a Brazilian city exposed to high rates of homicide

Cities in northeastern Brazil experience extreme rates of unsolved homicides, a situation that stimulates innovative procedures in the police work, such as forensic entomology. We surveyed necrophagous insects associated with carrion in a city exposed to high rates of homicides in Northeastern Brazil. The experiments were carried out in a rainforest fragment located in Recife, State of Pernambuco. Two pig carcasses were used as models, one in the dry and the other in the rainy season. The collection of adults was performed daily until the complete skeletonization of the carcasses. At least 32 Diptera species from the families Calliphoridae, Muscidae, Sarcophagidae, Fanniidae, Phoridae, Anthomyiidae, Piophilidae, and Stratiomyidae were registered, some of which have been previously documented on cadavers. A high richness of Diptera species was registered in all stages of decomposition. A strong overlap in the occurrence of most species was observed, which invalidates a defined entomological succession on the carcasses. Two species stood out in terms of abundance: Ophyra chalcogaster (Muscidae) and Chrysomya albiceps (Calliphoridae). The ubiquity of Hemilucilia semidiaphana (Calliphoridae) seems to confirm its preference for forest fragments exposed to low anthropogenic action. Our data contribute to expand the knowledge on the geographical distribution of forensically relevant species in the region and confirm the rapid dissemination of invasive Chrysomya species in forested areas.

Diversity of Diptera species associated with pig carcasses in a Brazilian city exposed to high rates of homicide

Cities in northeastern Brazil experience extreme rates of unsolved homicides, a situation that stimulates innovative procedures in the police work, such as forensic entomology. We surveyed necrophagous insects associated with carrion in a city exposed to high rates of homicides in Northeastern Brazil. The experiments were carried out in a rainforest fragment located in Recife, State of Pernambuco. Two pig carcasses were used as models, one in the dry and the other in the rainy season. The collection of adults was performed daily until the complete skeletonization of the carcasses. At least 32 Diptera species from the families Calliphoridae, Muscidae, Sarcophagidae, Fanniidae, Phoridae, Anthomyiidae, Piophilidae, and Stratiomyidae were registered, some of which have been previously documented on cadavers. A high richness of Diptera species was registered in all stages of decomposition. A strong overlap in the occurrence of most species was observed, which invalidates a defined entomological succession on the carcasses. Two species stood out in terms of abundance: Ophyra chalcogaster (Muscidae) and Chrysomya albiceps (Calliphoridae). The ubiquity of Hemilucilia semidiaphana (Calliphoridae) seems to confirm its preference for forest fragments exposed to low anthropogenic action. Our data contribute to expand the knowledge on the geographical distribution of forensically relevant species in the region and confirm the rapid dissemination of invasive Chrysomya species in forested areas.(AU)

Causes of death of sows in three Brazilian pig farms / Causas de morte de matrizes suínas em três granjas brasileiras

Sow mortality is directly related to financial losses and productive efficiency in pig farms. Despite this, diseases associated with the death of sows are poorly understood. This study aimed to determine the main causes of death of sows in Brazilian pig farms. To perform this research, three Brazilian pig farms were visited, and necropsies were performed on all sows that had died spontaneously or were subjected to euthanasia. Tissue fragments were collected for histopathological and bacteriological examination. In addition, the clinical signs, productive stage, parity, and type of death (spontaneous or euthanasia) were recorded. A total of 138 necropsies were performed, and 132 had a conclusive diagnosis. The most frequent productive stages were gestation and lactation (33.3 and 31.9%, respectively), followed by parturient sows (17.4%), cull sows (9.4%), weaning-to-estrus interval (WEI) (4.3%), and unmated gilts (3.6%). The most affected organ systems were reproductive (28%), digestive (25%), locomotor (22%), cardiovascular (9.1%), and hematopoietic (6.1%). The most frequently diagnosed conditions were uterine prolapse (16/132; 12.1%), gastric ulcer (13/132; 9.8%), suppurative arthritis (11/132; 8.3%), liver lobe torsion (11/132; 8.3%), heart failure (9/132; 6.8%), vaginal or vaginal and rectal prolapse (9/132; 6.8%), and pododermatitis (8/132; 6.1%). Although 58.2% of the deaths were due to one of these seven diseases, there was a great variability in diagnoses.(AU)

A mortalidade de porcas está diretamente relacionada a perdas financeiras e à eficiência produtiva das granjas. Apesar disso, as doenças associadas à morte de porcas são pouco conhecidas. Este estudo teve como objetivo determinar as principais causas de morte de porcas em granjas suinícolas brasileiras. Para a realização desta pesquisa, três granjas brasileiras foram visitadas e necropsias foram realizadas em todas as porcas que morreram espontaneamente ou foram submetidas à eutanásia. Fragmentos de tecidos foram coletados para exame histopatológico e bacteriológico. Além disso, foram registrados os sinais clínicos, estágio reprodutivo, ordem de parto e tipo de morte (espontânea ou eutanásia). Um total de 138 necropsias foram realizadas e 132 tiveram um diagnóstico conclusivo. Os estágios produtivos mais frequentes foram gestação e lactação (33,3 e 31,9%, respectivamente), seguidos por porcas parturientes (17,4%), porcas de descarte (9,4%), intervalo desmame-estro (IDE) (4,3%) e leitoas vazias (3,6%). Os sistemas orgânicos mais afetados foram reprodutor (28%), digestivo (25%), locomotor (22%), cardiovascular (9,1%) e hematopoiético (6,1%). As condições mais frequentemente diagnosticadas foram prolapso uterino (16/132; 12,1%), úlcera gástrica (13/132; 9,8%), artrite supurativa (11/132; 8,3%), torção do lobo hepático (11/132; 8,3%), insuficiência cardíaca (9/132; 6,8%), prolapso vaginal ou prolapso vaginal e retal (9/132; 6,8%) e pododermatite (8/132; 6,1%). Embora 58,2% dos óbitos tenham ocorrido por uma dessas sete doenças, houve grande variabilidade de diagnósticos.(AU)

Causes of death of sows in three Brazilian pig farms / Causas de morte de matrizes suínas em três granjas brasileiras

Sow mortality is directly related to financial losses and productive efficiency in pig farms. Despite this, diseases associated with the death of sows are poorly understood. This study aimed to determine the main causes of death of sows in Brazilian pig farms. To perform this research, three Brazilian pig farms were visited, and necropsies were performed on all sows that had died spontaneously or were subjected to euthanasia. Tissue fragments were collected for histopathological and bacteriological examination. In addition, the clinical signs, productive stage, parity, and type of death (spontaneous or euthanasia) were recorded. A total of 138 necropsies were performed, and 132 had a conclusive diagnosis. The most frequent productive stages were gestation and lactation (33.3 and 31.9%, respectively), followed by parturient sows (17.4%), cull sows (9.4%), weaning-to-estrus interval (WEI) (4.3%), and unmated gilts (3.6%). The most affected organ systems were reproductive (28%), digestive (25%), locomotor (22%), cardiovascular (9.1%), and hematopoietic (6.1%). The most frequently diagnosed conditions were uterine prolapse (16/132; 12.1%), gastric ulcer (13/132; 9.8%), suppurative arthritis (11/132; 8.3%), liver lobe torsion (11/132; 8.3%), heart failure (9/132; 6.8%), vaginal or vaginal and rectal prolapse (9/132; 6.8%), and pododermatitis (8/132; 6.1%). Although 58.2% of the deaths were due to one of these seven diseases, there was a great variability in diagnoses.(AU)

A mortalidade de porcas está diretamente relacionada a perdas financeiras e à eficiência produtiva das granjas. Apesar disso, as doenças associadas à morte de porcas são pouco conhecidas. Este estudo teve como objetivo determinar as principais causas de morte de porcas em granjas suinícolas brasileiras. Para a realização desta pesquisa, três granjas brasileiras foram visitadas e necropsias foram realizadas em todas as porcas que morreram espontaneamente ou foram submetidas à eutanásia. Fragmentos de tecidos foram coletados para exame histopatológico e bacteriológico. Além disso, foram registrados os sinais clínicos, estágio reprodutivo, ordem de parto e tipo de morte (espontânea ou eutanásia). Um total de 138 necropsias foram realizadas e 132 tiveram um diagnóstico conclusivo. Os estágios produtivos mais frequentes foram gestação e lactação (33,3 e 31,9%, respectivamente), seguidos por porcas parturientes (17,4%), porcas de descarte (9,4%), intervalo desmame-estro (IDE) (4,3%) e leitoas vazias (3,6%). Os sistemas orgânicos mais afetados foram reprodutor (28%), digestivo (25%), locomotor (22%), cardiovascular (9,1%) e hematopoiético (6,1%). As condições mais frequentemente diagnosticadas foram prolapso uterino (16/132; 12,1%), úlcera gástrica (13/132; 9,8%), artrite supurativa (11/132; 8,3%), torção do lobo hepático (11/132; 8,3%), insuficiência cardíaca (9/132; 6,8%), prolapso vaginal ou prolapso vaginal e retal (9/132; 6,8%) e pododermatite (8/132; 6,1%). Embora 58,2% dos óbitos tenham ocorrido por uma dessas sete doenças, houve grande variabilidade de diagnósticos.(AU)

Occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in pigs in the State of Pará, Brazil / Ocorrência de anticorpos contra Toxoplasma gondii e Neospora caninum em suínos no estado do Pará, Brasil

We investigated the occurrence of Toxoplasma gondii and Neospora caninum antibodies in pigs raised in the Northeast of Pará, Brazil. At Study I, convenience sampled 151 pigs at two slaughterhouses, with and without state inspection; and Study II, which assessed 159 pigs with probabilistic sampling from nine pig farms. Serological analysis was performed using indirect fluorescent antibody test for T. gondii and N. caninum with a cutoff of 64 and 50, respectively. Overall, 6.77% pigs were seropositive for T. gondii and 5.16% for N. caninum. In Study I, pigs slaughtered with and without state inspection presented similar occurrence for both coccidia (p>0.05). Study II found an association between N. caninum seropositivity and sludge discarded into the soil, feeding pigs with animal-based protein, subsistence system, and absence of nipple drinkers. No association was found for T. gondii. Pigs from Pará are a potential source of T. gondii infection to humans. To our best knowledge, this is the first study to report anti-N. caninum antibodies in the serum of pigs in Pará State, Brazilian Amazon.(AU)

Foi investigada a ocorrência de anticorpos contra Toxoplasma gondii e Neospora caninum em suínos criados no nordeste do Pará, Brasil. No Estudo I, foram amostrados 151 porcos em dois matadouros, com e sem inspeção estadual. O Estudo II avaliou 159 suínos com amostragem probabilística de nove granjas de suínos. Para sorologia, utilizou-se o teste de imunofluorescência indireta para T. gondii e N. caninum com ponto de corte de 1:64 e 1:50, respectivamente. No geral, 6,77% dos suínos foram soropositivos para T. gondii e 5,16% para N. caninum. No Estudo I, suínos abatidos em matadouros com e sem inspeção estadual apresentaram ocorrência semelhante para ambos os coccídios (p> 0,05). Os animais amostrados de Belém, Benevides, Marituba, Bujaru, Castanhal e Igarapé-Miri foram positivos para T. gondii, enquanto os soropositivos para N. caninum foram encontrados em Belém, Bujaru, Castanhal e Santo Antônio do Tauá. O Estudo II encontrou associação entre soropositividade de N. caninum e esterco descartado no solo, alimentação dos suínos com proteína de origem animal, criação de subsistência e ausência de bebedores tipo nipple. Não foi encontrada associação para T. gondii. A carne suína apresenta potencial risco de transmissão de T. gondii para os habitantes da região. De acordo com nosso conhecimento, este é o primeiro relato de anticorpos anti-N. caninum em suínos no estado do Pará, Amazônia brasileira.(AU)

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production

The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.(AU)

Producers' knowledge and perception about environmental enrichment and materials used in pig farms

This study investigated the knowledge and perception of Brazilian pig producers in terms of environmental enrichment (EE) for pigs, the materials used, and the forms of presentation, identifying the single conditions that can improve their application on pig farms. A questionnaire was applied to 1340 farms ­ representing 7.4% of the farms and 12% of the sows in the country. The questions included descriptions of farms and respondents, their knowledge, judgments and level of interest in the use of EE, and frequency of use of the materials. Enrichment was used in 89.1% of farms, but half of the respondents admitted to using it without knowing what was involved. The producers presented an optimistic view (92.3%) about the application of this tool and were interested in receiving more knowledge and guidance on the subject (97.8%). The materials were used mainly to avoid fights (46.3%) and to prevent tail biting (23.3%); on the other hand, the increase in production costs (39.6%) and lack of knowledge about the subject (31.3%) were the main reasons for not using enrichment; concerns about increased management on farms were sporadic (7.1%). Metal chains were the main artifacts used, followed by plastic containers and pieces of wood; the same breeding farm used up to five types of materials. The fact that the respondents stated that they knew what EE was and that they had a good perspective on its use were significant conditions for the use of some kind of enrichment on the farms. Younger farm owners were more likely to use enrichment materials than more experienced ones. The results suggested that Brazilian pig producers use EE even with limited knowledge about the subject and that there is an argument to improve the use of this animal welfare strategy.(AU)

Antimicrobial resistance in bacteria isolated from pigs with respiratory clinical signs in Brazil / Resistência a antimicrobianos em bactérias isoladas de suínos com sinais clínicos respiratórios no Brasil

Antimicrobial resistance is a current and important issue to public health, and it is usually associated with the indiscriminate use of antimicrobials in animal production. This study aimed to evaluate the antimicrobial susceptibility profile in bacterial isolates from pigs with clinical respiratory signs in Brazil. One hundred sixty bacterial strains isolated from pigs from 51 pig farms in Brazil were studied. In vitro disk-diffusion method was employed using 14 antimicrobial agents: amoxicillin, penicillin, ceftiofur, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, oxytetracycline, tetracycline, erythromycin, tilmicosin, florfenicol, lincomycin, and sulfadiazine/trimethoprim. The majority of isolates were resistant to at least one antimicrobial agent (98.75%; 158/160), while 31.25% (50/160) of the strains were multidrug resistant. Streptococcus suis and Bordetella bronchiseptica were the pathogens that showed higher resistance levels. Haemophilus parasuis showed high resistance levels to sulfadiazine/trimethoprim (9/18=50%). We observed that isolates from the midwestern and southern regions exhibited four times greater chance of being multidrug resistant than the isolates from the southeastern region studied. Overall, the results of the present study showed a great level of resistance to lincomycin, erythromycin, sulfadiazine/trimethoprim, and tetracycline among bacterial respiratory pathogens isolated from pigs in Brazil. The high levels of antimicrobial resistance in swine respiratory bacterial pathogens highlight the need for the proper use of antimicrobials in Brazilian pig farms.(AU)

A resistência antimicrobiana é uma questão atual e muito importante para a saúde pública, geralmente associada ao uso indiscriminado de antimicrobianos na produção animal. Diante disso, foi investigado o perfil de sensibilidade-antimicrobiana em isolados bacterianos de suínos com sinais clínicos respiratórios no Brasil. Foram estudadas 96 isolados provenientes de 51 granjas de suínos do Brasil. O método de disco-difusão foi empregado usando 14 antimicrobianos: amoxicilina, penicilina, ceftiofur, ciprofloxacina, enrofloxacina, clortetraciclina, doxiciclina, oxitetraciclina, tetraciclina, eritromicina, tilmicosina, florfenicol, lincomicina e sulfadiazina/trimetoprim. Streptococcus suis e Bordetella bronchiseptica foram os patógenos que apresentaram maiores níveis de resistência. Haemophilus parasuis apresentou altos níveis de resistência à sulfadiazina/trimetoprim (9/18=50%). Observou-se que isolados das regiões Centro-Oeste e Sul apresentaram quatro vezes mais chance de serem multirresistentes do que os isolados da região Sudeste. A maioria foi resistente a pelo menos um agente antimicrobiano (98,75%; 158/160) e 31,25% (50/160) das estirpes isoladas eram multirresistentes. No geral, os resultados do presente estudo mostraram grande nível de resistência à lincomicina, eritromicina, sulfadiazina/trimetoprim e tetraciclina entre patógenos respiratórios bacterianos isolados de suínos no Brasil. Os altos níveis de resistência antimicrobiana em patógenos bacterianos respiratórios em suínos reforçam a necessidade do uso criterioso de antimicrobianos na suinocultura brasileira.(AU)