Detection of anti-Toxoplasma gondiiantibodies in wild free-living birds and mammals from the northwest region of São Paulo state, Brazil / Detecção de anticorpos anti-Toxoplasma gondiiem aves e mamíferos silvestres de vida livre da região noroeste do estado de São Paulo, Brasil

Toxoplasmosis is a protozoonosis caused by an obligate intracellular parasite named Toxoplasma gondii, which can infect humans and a large number of homeothermic animal species with worldwide distribution. The present study aimed to detect anti-T. gondii antibodies from serological samples of free-living wild animals from the northwest region of São Paulo state, Brazil. Thirty-two samples (eight from birds and 24 from mammals) were analyzed by the modified agglutination test (MAT) using 5 cut-off points for birds and 25 for mammals. Seropositivity was observed in 25% (2/8) of birds, including the species Rupornis magnirostris (roadside hawk) and Caracara plancus (southern caracara), and 29.2% (7/24) animals were seropositive among mammals, including one hoary fox (Lycalopex vetulus), two maned wolves (Chrysocyon brachyurus), one black howler monkey (Alouatta caraya), two crab-eating foxes (Cerdocyon thous) and one gray brocket deer (Mazama gouazoubira). The results obtained with the present study indicate the exposure to T. gondiiof free-living wild animals from the northwest region of São Paulo state and, therefore, that they probably play a role in the transmission and maintenance of T. gondii in the environment they inhabit. Thus, identification of the infection in several animal species in the region indicates the environmental contamination of the area. Studies of this nature may help to understand the importance of the prevention and control of this disease in Brazil.(AU)

A toxoplasmose é uma protozoonose causada por um parasita intracelular obrigatório denominado Toxoplasma gondii, que pode infectar os humanos e um vasto número de espécies animais homeotérmicas, apresentando distribuição mundial. O presente estudo objetivou a detecção de anticorpos anti-T. gondii a partir de amostras sorológicas de animais silvestres de vida livre da região noroeste do estado de São Paulo. Foram analisadas 32 amostras (oito de aves e 24 de mamíferos) por meio do teste de aglutinação modificado (MAT), utilizando ponto de corte 5 para as aves e 25 para os mamíferos. Soropositividade foi observada em 25% (2/8) das aves, incluindo as espécies Rupornis magnirostris (gavião-carijó) e Caracara plancus (carcará); entre os mamíferos, 29,2% (7/24) foram soropositivos incluindo uma raposa-do-campo (Lycalopex vetulus), dois lobos-guará (Chrysocyon brachyurus), um bugio-preto (Alouatta caraya), dois cachorros-do-mato (Cerdocyon thous) e um veado-catingueiro (Mazama gouazoubira). Os resultados obtidos com o presente estudo indicam a exposição dos animais selvagens de vida livre a T. gondii na região noroeste do estado de São Paulo e, portanto, que provavelmente apresentam papel na transmissão e manutenção de T. gondii no meio ambiente em que vivem. Assim, a identificação da infecção em várias espécies de animais na região indica a contaminação ambiental da área. Estudos dessa natureza podem ajudar no entendimento sobre a prevenção e o controle dessa importante doença no Brasil.(AU)

Ocorrência de anticorpos anti-toxoplasma gondii em aves silvestres de três unidades de conservação federais da Paraíba e da Bahia / Occurrence of antibodies to toxoplasma gondii in wild birds from three federal conservation units of Paraíba and Bahia, Brazil

A vigilância e monitoramento de doenças em animais silvestres são imprescindíveis no contexto ambiental e de saúde pública, pois estes animais agem como sentinelas, refletindo alterações ambientais precocemente, o que proporciona maior eficácia no monitoramento ambiental e permite o acesso rápido a informações sobre as condições da área. Neste contexto, as aves são importantes no ciclo biológico do Toxoplasma gondii e na epidemiologia da toxoplasmose, principalmente porque seus tecidos representam importantes fontes de proteína na alimentação de felídeos e humanos. Objetivou-se detectar anticorpos anti-T. gondii, por meio do teste de aglutinação modificada em aves silvestres de três Unidades de Conservação (UC) Federais dos Estados da Paraíba e Bahia. No período de dezembro de 2011 a outubro de 2013 foram capturadas com redes de neblina 222 aves silvestres pertencentes a 67 espécies, 27 famílias e 12 ordens. Após a captura, foi colhido sangue de cada animal e separado o soro, que foi submetido ao Teste de Aglutinação Modificada (MAT≥1:25) utilizando taquizoítos inativados na formalina e 2-mercaptoetanol. Dentre as 222 amostras analisadas, três (1,3%) foram sororreagentes: 1 de 16 (6,2%) pipira-preta Tachyphonus rufus (título 50), 1 de 5 (20%) juriti-gemedeira Leptotila rufaxilla (título 50) e 1 de 1 (100%) caneleiro-enxofre Casiornis fuscus (título 25). Este é o primeiro relato da ocorrência de anticorpos anti-T. gondii nas referidas espécies de aves silvestres de vida livre nas duas UC Federais estudadas.

Surveillance and monitoring of wildlife pathogens are essential in the environmental context and human public health, as these animals act as sentinels, reflecting environmental changes early on, whath gives more efficient environmental monitoring and allows quick access to information on the conditions of area. Birds are important in the epidemiology and life cycle of Toxoplasma gondii, because their tissues are important source of protein in the diet of felids and humans. The objective was to determine antibodies to Toxoplasma gondii in wild birds from three Federal Conservation Units of the states of Paraíba and Bahia by Modified Agglutination Test (MAT). From December 2011 to October 2013, 222 wild birds of 67 species from 27 families and 12 Orders were captured with mist nets. Blood samples were then collected and the serum was separated by centrifugation. The sera were tested (MAT≥1:25) using formalin-fixed whole tachyzoites and 2-mercaptoethanol. Antibodies to T. gondii were found in 3 of 222 (1.3%) birds: in 1 of 16 (6.2%) white-lined tanager (Tachyphonus rufus, titer 50), in 1 of 5 (20%) gray-fronted dove (Leptotilla rufaxila, titer 50), and in 1 of 1 (100%) ashy-throated casiornis (Casiornis fuscus, titer 25). This is the first report of occurrence of antibodies to T. gondii in these tree bird species from two Federal Conservation Units.

Protective roles of free avian respiratory macrophages in captive birds

In the mammalian lung, respiratory macrophages provide front line defense against invading pathogens and particulate matter. In birds, respiratory macrophages are known as free avian respiratory macrophages (FARM) and a dearth of the cells in the avian lung has been purported to foreordain a weak first line of pulmonary defense, a condition associated with high mortality of domestic birds occasioned by respiratory inflictions. Avian pulmonary mechanisms including a three tiered aerodynamic filtration system, tight epithelial junctions and an efficient mucociliary escalator system have been known to supplement FARM protective roles. Current studies, however, report FARM to exhibit an exceptionally efficient phagocytic capacity and are effective in elimination of invading pathogens. In this review, we also report on effects of selective synthetic peroxisome proliferator activated receptor gamma (PPAR γ) agonists on non phlogistic phagocytic properties in the FARM. To develop effective therapeutic interventions targeting FARM in treatment and management of respiratory disease conditions in the poultry, further studies are required to fully understand the role of FARM in innate and adaptive immune responses.

Pet avoidance in allergy cases: Is it possible to implement it? / Evitación de mascotas en casos de alergia. ¿Es factible lograrla?

Introduction: Among allergic patients, pet avoidance is commonly recommended. It is difficult for patients to accomplish this because of their emotional attachment to the pets, and its effectiveness is controversial. Objective: To explore the applicability and effectiveness of pet avoidance measures among sensitized patients. Materials and methods: We evaluated 288 patients with asthma, rhinitis, conjunctivitis and/or dermatitis using skin prick test to measure their sensitization to cats, dogs and other animals to which they were exposed. Exposure to animals was evaluated in each patient (pets at home, frequent indirect exposure or no exposure). In those patients sensitized to animals some avoidance measures, such as removing pets from home and preventing indirect exposure, were recommended. On the following two appointments, we evaluated patients' fulfillment of these recommendations. Results: Sensitization to cats, dogs and birds was high (9%, 48%, 14%, respectively), as well as direct and indirect exposure (30%, 46%, 24%, respectively). Most patients denied contact with other animals (horses, hamsters, rabbits or cows), and sensitization to them was low. During the follow-up of patients sensitized to their pets at home (n=50), most of them refused to remove them from their house due to emotional attachment, and only two followed this recommendation. Conclusions: High exposure to animals could explain the frequency of sensitization to pets in this population. However, emotional attachment and prevalent indirect exposure to animals among sensitized patients make avoidance recommendations impractical or impossible to achieve.

Introducción. Entre los pacientes alérgicos se recomienda comúnmente la evitación de mascotas; sin embargo, es difícil que los pacientes cumplan con esta recomendación debido al apego emocional y, además, su efecto clínico no es claro. Objetivo. Explorar la aplicabilidad de las medidas de evitación entre pacientes sensibilizados a mascotas . Materiales y métodos. En 284 pacientes con asma, rinitis, conjuntivitis y dermatitis, se evaluó la sensibilización a gatos, perros y otros animales mediante pruebas de punción epidérmica. Se evaluó, igualmente, el nivel de exposición a animales (mascotas en la casa y exposición indirecta frecuente). A aquellos pacientes sensibilizados a los animales, se les recomendaron medidas de evitación como retirar la mascota de la casa y evitar la exposición indirecta. En las dos citas médicas siguientes se evaluó el cumplimiento de estas recomendaciones. Resultados. La sensibilización a gatos, perros y aves fue alta (9, 48, y 14 %, respectivamente), al igual que la exposición directa o indirecta a estos animales (30, 46, 24 %, respectivamente). La mayoría de los pacientes negó el contacto frecuente con otros animales (caballos, hámsteres, conejos, vacas) y la sensibilización a estos fue baja. La mayoría de los pacientes sensibilizados a su propia mascota (n= 50) se rehusó a retirar la mascota de la casa y solo dos de ellos siguieron la recomendación de hacerlo. Conclusiones. La exposición frecuente a los animales podría explicar la gran frecuencia de sensibilización a las mascotas en esta población. Sin embargo, el apego emocional y la exposición indirecta frecuente, hacen que las recomendaciones de evitación sean imprácticas o casi imposibles de lograr.

Seleção, caracterização e clonagem dos genes fljB e groEL agonistas dos receptores de reconhecimento de padrão do sistema imune inato das aves / Selection, characterization and cloning of the fljB and groEL genes agonists of the innate immune system pattern recognition receptors of birds

The recombinant production of innate immune system pattern recognition receptor agonists has provided a new tool for the production of immunostimulants for animals. The molecular pattern associated with the pathogen (PAMP), flagellin, coded by the fljB gene from Salmonella Typhimirium, and the molecular pattern associated to the damage (DAMP), HSP60, coded by the groEL gene from S. Typhimurium and S. Enteritidis, are recognized by pattern recognition receptors (PRRs) of the innate immune system of birds. In the present study, we performed the cloning of genetic fragments of the genes fljB, from S. Typhimurium, and groEL from S. Typhimurium and S. Enteritidis inserted in expression vector pET100/D-TOPO and transformed in E. coli TO10 cells. The clones were evaluated by colony PCR, plasmidial DNA PCR and genome sequencing in order to confirm the presence of these genes. In the colony PCR, we identified the presence of genes groEL (S. Enteritidis), groEL (S. Typhimurium) and fljB (S. Typhimurium) in 80%, 60% and 80% of the transformed colonies, respectively. The cloning system adopted allowed the production of HSP60 genetic fragment clones and flagellin of Salmonella strains, allowing the posterior use of these clones in gene expression trials, with the future potential of being used as non-specific immunostimulants for birds.

A produção recombinante de agonistas dos receptores do reconhecimento de padrão do sistema imune inato tem fornecido uma nova ferramenta para a produção de imunoestimulantes para animais. O padrão molecular associado ao patógeno (PAMP), flagelina, codificado pelo gene fljB de Salmonella Typhimurium e o padrão molecular associado ao dano (DAMP) HSP60, codificado pelo gene groEL da S. Typhimurium e S. Enteritidis, são reconhecidos por receptores de reconhecimento de padrões (RRPs) do sistema imune inato das aves. No presente estudo, foi feita a clonagem de fragmentos genéticos dos genes fljB de S. Typhimurium e groEL de S. Typhimurium e S. Enteritidis inseridos no vetor de expressão pET100/D-TOPO e transformados em células de E. coli TOP10. Os clones foram avaliados pela PCR de colônia, PCR de DNA plasmidial e sequenciamento genômico para a confirmação da presença desses genes. Na PCR de colônia, foram identificadas em 80%, 60% e 80% das colônias transformadas, a presença dos genes groEL (S. Enteritidis), groEL (S. Typhimurium) e fljB (S. Typhimurium) respectivamente. O sistema de clonagem adotado possibilitou a produção de clones dos fragmentos genéticos da HSP60 e flagelina das cepas de Salmonella, permitindo a utilização posterior desses clones em ensaios de expressão gênica, com potencial futuro de serem utilizados como imunoestimulante inespecífico das aves.

Comparison of techniques of detecting immunoglobulin-binding protein reactivity to immunoglobulin produced by different avian and mammalian species / Comparación de las técnicas de detección de la reactividad de la proteína de unión de la inmunoglobulina frente a la inmunoglobulina producida por diferentes especies aviarias y mamíferas

The rationale of this study was to use several immunological assays to investigate the reactivity of immunoglobulin binding protein (IBP) to immunoglobulins from various avian and mammalian species. The IBP studied were Staphylococcal protein A (SpA), Streptococcal protein G (SpG), Peptostreptococcal protein L (SpL) and recombinant protein LA (SpLA). The various immunological techniques used were double immunodiffusion (Ouchterlony technique) that tested positive high protein reactivities, direct and competitive enzyme-linked immunosorbent assays (ELISAs) that tested moderate and low positive protein binding capacities, respectively. In addition to sandwich ELISAs, immunoblot analyses and Ig-purification by SpA-affinity chromatography, which were sensitive tests and helpful in the screening and confirmatory tests were also used. The Ouchterlony technique showed that compared to the other proteins, SpLA had the highest range of reactivity with animal sera and purified immunoglobulins while SpL was least reactive. With the direct ELISA, SpL reacted with the raccoon sera, rabbit IgG and with IgY from bantam hens and pigeons. While with the direct ELISA, SpA reacted with sera from skunk, coyote, raccoon, mule, donkey and human. The sandwich ELISA revealed high reactivity of both SpG and SpLA with mammalian sera titres ranging from 1:32 (raccoon serum) to 1:1024 (mule and donkey sera).These results suggest that IBP can be used for the detection of immunoglobulin using various immunological assays and this is important for the diagnosis of infectious diseases in animal and bird populations studied and in the purification of immunoglobulins.

El fundamento de este estudio radica en el uso de varios ensayos inmunológicos para investigar la reactividad de la proteína de unión de la inmunoglobulina (IBP) frente a las inmunoglobulinas de varias especies aviarias y mamíferas. Las proteínas IBP estudiadas fueron la proteína estafilocócica A (SpA), la proteína estreptocócica G (SpG), la proteína peptoestreptocócica L (SpL), y la proteína recombinante LA (SpLA). Las varias técnicas inmunológicas usadas fueron: la inmunodifusión doble (técnica de Ouchterlony) para examinar las reactividades positivas de la proteína alta; el ensayo por inmunoabsorción ligado a enzimas(ELISA), de tipo directo y competitivo, para examinar la capacidad de realizar uniones positivas de proteína moderada y baja, respectivamente, además del ensayo ELISA 'Sándwich', los análisis inmunoblot, yla purificación de IgG, mediante cromatografía de afinidad, los cuales fueron pruebas sensibles y útiles en el tamizaje y las pruebas de confirmación. La técnica de Ouchterlony mostró que - en comparación con otras proteínas - la SpLA tenía el grado más alto de reactividad con los sueros animales y las inmunoglobulinas purificadas, mientras que la SpL fue la menos reactiva. Con el ELISA directo, la SpL reaccionó con los sueros de mapache, la IgG de conejo, así como con la IgY de palomas y gallinas de Bantam, en tanto con el ELISA directo, la SpA reaccionó con sueros de mofeta, coyote, mapache, mula, asno y seres humanos. ELISA "sándwich" reveló una alta reactividad tanto de SpG como de SpLA, con títulos séricos mamíferos que iban desde 1:32 (suero de mapache) hasta 1:1024 (sueros de mula y de asno). Estos resultados sugieren que la proteína de unión IBP puede usarse en la detección de la inmunoglobulina usando varios ensayos inmunológicos, lo cual es importante para el diagnóstico de enfermedades infecciosas en las poblaciones animales y aviarias bajo estudio, así como para la purificación de inmunoglobulinas.

Follow-up of the development of femoral degeneration lesions in broilers / Seguimiento del desarrollo de lesiones por degeneración en pollos de engorde

A study was carried out in the experimental facilities of FMVZ/UNESP-Botucatu, with the aim of following-up the development and the incidence of femoral degeneration (FD). A total of 305 one-day-old male broilers were housed in six pens of 5m² each. Histological analyses of femur head collected when broilers were 0, 7, 14, 21, 28, 35, and 42 days of age were carried out. At 42 days of age, 30 birds were taken to the experimental processing plant of FMVZ for leg gross examination. Ten legs per FD score where selected, and histologically analyzed to determine the most probable age at the beginning of the lesions, and to standardize femoral degeneration lesion scores. The histological results showed that cell architecture started to disorganize at 21 days of age in the resting and proliferation zones, and that angiogenesis increased, invading the joint cartilage, The gross lesion indexes due to femoral degeneration were 22.5 percent, 42.5%, and 65% at 28, 35, and 42 days of age, respectively.

Se realizó un estudio en las instalaciones experimentales de FMVZ/UNESP-Botucatu, con el objetivo de seguir el desarrollo y la incidencia de degeneración femoral (DF) en pollos. Se utilizaron 305 polluelos de un día, machos, distribuidos en seis corrales de 5m² cada uno. Se analizaron cortes histológicos de cabezas de fémur recolectadas a los 0, 7, 14, 21, 28, 35 y 42 días de edad. A los 40 días de edad, se llevaron 30 aves al Matadero Experimental de FMVZ, para análisis macroscópico de las piernas. Se escogieron 10 muslos por escore de DF, y se analizaron histológicamente para determinar la edad más probable del inicio de la lesión y estandarizar los escores de lesión por degeneración femoral. Los resultados histológicos indicaron que a los 21 días ocurre el inicio de la desorganización celular en la zona de reposo y de proliferación, además del aumento de la angiogénesis, invadiendo el cartílago articular. Microscópicamente, el índice de lesión por degeneración femoral fue del 22.5 por ciento, 42.5% y 65% a los 28, 35 y 42 días de edad, respectivamente.

Consideraciones técnicas sobre la problemática de las micotoxinas y las micotoxicosis aviares / Technical considerations on the problem of mycotoxins and poultry mycotoxicosis

La contaminación con Micotoxinas puede ocurrir en el cultivo, cosecha, almacenaje, incluso en el alimento producido. Las Micotoxinas son compuestos altamente estables que pueden producir diversos cuadros en animales y en el ser humano. En las aves, las Micotoxinas más importantes son Aflatoxina, Ocratoxina, Toxina T-2, Diacetoxiscirpenol (DAS), Fumoninisina, Citrinina y Ácido Ciclopiazónico. Entre los mecanismos de prevención de la producción de las Micotoxinas —que comienzan con la eliminación o disminución del crecimiento de hongos—, resalta que en el almacenaje se recomienda utilizar inhibidores de hongos, y uso de agentes secuestrantes o absorbentes de Micotoxinas.

Anticuerpos aviares: alternativa en producción y diagnóstico / Avian antibodies: an alternative in production and diagnosis

El uso de anticuerpos para investigación y diagnóstico se realiza desde hace varias décadas en todo el mundo. Normalmente, estos anticuerpos se obtienen a partir del suero de mamíferos (roedores, caprinos, equinos, etc.) De acuerdo con el tamaño de huésped, se pueden producir pequeñas o grandes cantidades de suero, haciendo siempre sangrías regulares para su recolección. En los últimos años, se han utilizado cada vez con mayor frecuencia anticuerpos purificados a partir de huevos de gallina inmunizadas, los cuales presentan diferencias con los anticuerpos producidos en mamíferos en su estructura y características fisicoquímicas, pero además, son una alternativa que disminuye el estrés e injuria al huésped y tiene alta productividad y facilidad para su recolección. Se ha informado el uso de estos anticuerpos aviares en ensayos inmunoquímicos, producción de conjugados y en terapéutica con un éxito similar al de los anticuerpos en mamíferos y a un costo menor. En este trabajo, se hace una revisión del tema y se plantean sus posibles usos tanto en investigación y diagnóstico, como en terapia

Técnicas de medición de estrés en aves / Stress measuring techniques used in birds

Las condiciones ambientales que presentan los sitemas de producción intensivos propician estado de estrés en los animales domésticos. La producción comercial de aves es uno de los principales ejemplos de lo anterior, ya que cada día se hace más patente la relación de la presencia de problemas de salud y producción que pueden tener relación con los niveles de estrés al que se encuentra sometida esta especie. La medición sistemática de los niveles de estrés al que se encuentra sometida esta especie. La medición sitemática de los niveles de estrés es uno de los primeros pasos para su determinación objetiva y las consecuencias en la productividad animal. La presente revisión muestra algunas de las técnicas utilizadas en aves para llevar a cabo estas mediciones, unas de tipo directo como las conductuales y la prueba de ACTH exógena, y otras de tipo indirecto como pruebas hematológicas, medición de glucagon pancreático y la interpretación de los resultados en los parámetros de producción

Adecuación de la técnica de separación de heterófilos y monocitos aviares de sangre completa, utilizando gradientes discontínuos de Ficoll-Hypaque / Standardization of separation of avian heterophils and monocytes from whole blood using Ficoll-Hypaque discontinuous gradients

Se realizó la adecuación de la técnica de gradientes discontinuos de Ficoll-Hypaque, para la obtención de monocitos y heterófilos aviares puros y viables a partir sangre completa de pollos sanos de entre 8 y 9 semanas de edad. Al final de 10 repeticiones la cuenta total de monocitos obtenida fue de 0.25 X 10 6 a 0.46 X 10 6 por ml, con un porcentaje promedio de viabilidad y pureza de 97.65 ñ 1 para la primera y 95.17 ñ 0.97 para la segunda. La cuenta total de heterófilos fue de 0.41 X 10 6 a 1.15 X 10 6 por ml, con un porcentaje promedio de viabilidad y pureza de 99.4 ñ 0.4 para la primera y 96.77 ñ 1.88 para la segunda. Mediante esta técnica se pueden obtener monocitos y heterófilos viables, puros y cultivos in vitro para determinar mecanismos de quimiotaxis, fagocitosis y lisis contra bacterias

Relación entre mosquitos vectores y aves acuáticas en la transmisión potencial de dos arbovirus / The relationship between mosquito vectors and aquatic birds in the potential transmission of 2 arboviruses

The authors studied for two years the role of the chicks of aquatic birds in the arboviral cycles in coastal lagoons in central Panama in order to determine the relation between Culex (Melanoconion) ocossa and Mansonia (Mansonia) dyari mosquitoes in the transmission and dissemination of the viruses of Saint Louis Encephalitis (SLE) and Venezuelan Equine Encephalitis (VEE). Mosquitoes were captured every fifteen days on two consecutive nights to isolate the virus, using light traps (CDC) and baited traps. The attempts to isolate the virus were made using Vero cell cultures and the determination of antibodies was performed. The results of the serologic tests seem to indicate that four bird species: the ex (?) heron (Bubulcus ibis), the American heron (Casmerodius albus), the spoon-billed duck (Cochlearius cochlearius) and the needle crow (Anhinga anhinga) could function as intermediate hosts in the transmission cycle of SLE. Two species, the ibis (Endocimus albus) and the spoon-billed duck (Cochlearius cochlearius) could also be intermediate hosts of VEE in the coastal lagoons of Panama. The presence of antibodies in chicks could indicate an infection acquired recently, after their birth, in this area. The VEE virus was recovered from blood filled mosquitoes which had fed on a spoon-billed duck probably infected and exposed in a Trinidad #10 trap. No SLE virus was isolated. Other unknown viruses were isolated from mosquitoes selected for these studies, such as C. ocossa and M. dyari. The results obtained with these studies indicate the need for more studies utilizing new field techniques in order to establish a link between SLE and VEE, the vector mosquitoes and the aquatic birds in the coastal lagoons of the area under investigation