Detection of the ADP-ribosyltransferase toxin gene (cdtA) and its activity in Clostridium difficile isolates from Equidae

Clostridium difficile is an antibiotic-associated emerging pathogen of humans and animals. Thus far three toxins of C. difficile have been described: an enterotoxin (ToxA), a cytotoxin (ToxB) and an ADP-ribosyltransferase (CDT). In the present work we describe the first isolation of CDT producing C. difficile from Equidae with gastro-intestinal disease. Out of 17 C. difficile strains isolated from Equidae, 11 were positive for the genes tcdA and tcdB encoding ToxA and ToxB. In addition four of these 11 isolates were positive for the cdtA gene encoding the catalytic subunit of the ADP-ribosyltransferase CDT. Interestingly none of the isolates derived from canines (41 isolates) and felines (4 isolates) harboured the cdtA gene. In C. difficile field isolates which contained the cdtA gene, ADP-ribosyltransferase activity could also be detected in culture supernatants indicating expression and secretion of CDT. All strains were associated with intestinal disorders, but no association was found for the occurrence of toxins with a specific clinical diagnosis.

IL26 gene inactivation in Equidae

Interleukin-26 (IL26) is a member of the IL10 cytokine family. The IL26 gene is located between two other well-known cytokines genes of this family encoding interferon-gamma (IFNG) and IL22 in an evolutionary conserved gene cluster. In contrast to humans and most other mammals, mice lack a functional Il26 gene. We analyzed the genome sequences of other vertebrates for the presence or absence of functional IL26 orthologs and found that the IL26 gene has also become inactivated in several equid species. We detected a one-base pair frameshift deletion in exon 2 of the IL26 gene in the domestic horse (Equus caballus), Przewalski horse (Equus przewalskii) and donkey (Equus asinus). The remnant IL26 gene in the horse is still transcribed and gives rise to at least five alternative transcripts. None of these transcripts share a conserved open reading frame with the human IL26 gene. A comparative analysis across diverse vertebrates revealed that the IL26 gene has also independently been inactivated in a few other mammals, including the African elephant and the European hedgehog. The IL26 gene thus appears to be highly variable, and the conserved open reading frame has been lost several times during mammalian evolution.

In Plant Activation: An inducible, hyperexpression platform for recombinant protein production in plants

In this study, we describe a novel protein production platform that provides both activation and amplification of transgene expression in planta. The In Plant Activation (INPACT) system is based on the replication machinery of tobacco yellow dwarf mastrevirus (TYDV) and is essentially transient gene expression from a stably transformed plant, thus combining the advantages of both means of expression. The INPACT cassette is uniquely arranged such that the gene of interest is split and only reconstituted in the presence of the TYDV-encoded Rep/RepA proteins. Rep/RepA expression is placed under the control of the AlcA:AlcR gene switch, which is responsive to trace levels of ethanol. Transgenic tobacco (Nicotiana tabacum cv Samsun) plants containing an INPACT cassette encoding the b-glucuronidase (GUS) reporter had negligible background expression but accumulated very high GUS levels (up to 10% total soluble protein) throughout the plant, within 3 d of a 1% ethanol application. The GUS reporter was replaced with a gene encoding a lethal ribonuclease, barnase, demonstrating that the INPACT system provides exquisite control of transgene expression and can be adapted to potentially toxic or inhibitory compounds. The INPACT gene expression platform is scalable, not host-limited, and has been used to express both a therapeutic and an industrial protein.

The evolution and biogeography of the austral horse fly tribe Scionini (Diptera: Tabanidae: Pangoniinae) inferred from multiple mitochondrial and nuclear genes

Phylogenetic relationships within the Tabanidae are largely unknown, despite their considerable medical and ecological importance. The first robust phylogenetic hypothesis for the horse fly tribe Scionini is provided, completing the systematic placement of all tribes in the subfamily Pangoniinae. The Scionini consists of seven mostly southern hemisphere genera distributed in Australia, New Guinea, New Zealand and South America. A 5757. bp alignment of 6 genes, including mitochondrial (COI and COII), ribosomal (28S) and nuclear (AATS and CAD regions 1, 3 and 4) genes, was analysed for 176 taxa using both Bayesian and maximum likelihood approaches. Results indicate the Scionini are strongly monophyletic, with the exclusion of the only northern hemisphere genus Goniops. The South American genera Fidena, Pityocera and Scione were strongly monophyletic, corresponding to current morphology-based classification schemes. The most widespread genus Scaptia was paraphyletic and formed nine strongly supported monophyletic clades, each corresponding to either the current subgenera or several previously synonymised genera that should be formally resurrected. Molecular results also reveal a newly recognised genus endemic to New Zealand, formerly placed within Scaptia. Divergence time estimation was employed to assess the global biogeographical patterns in the Pangoniinae. These analyses demonstrated that the Scionini are a typical Gondwanan group whose diversification was influenced by the fragmentation of that ancient land mass. Furthermore, results indicate that the Scionini most likely originated in Australia and subsequently radiated to New Zealand and South American by both long distance dispersal and vicariance. The phylogenetic framework of the Scionini provided herein will be valuable for taxonomic revisions of the Tabanidae.

Factibilidad productiva y económica de producción equina para carne

Resumen: El propósito del siguiente trabajo es colaborar al esclarecimiento de una producción poco conocida y desarrollada en nuestro país, la cual a pesar de su condición permite ingresar anualmente más de 70 millones de dólares. La producción de equinos para carne propiamente dicha no existe, la faena nacional se abastece de animales descartados, lesionados o viejos. Para poder determinar la viabilidad de esta producción, se recurrió al aporte de datos, por medio de un análisis de la producción equina, descripción de sistemas productivos, clasificación y cuantificación de costos e ingresos, ventajas y desventajas de la producción, evaluación del mercado y legislación vigente. Finalizado este análisis, queda demostrado que el territorio nacional posee la capacidad para producir y albergar equinos con destino exclusivo a faena. Por otro lado, el análisis económico parcial realizado evidencia la escasez del margen, donde el precio del kilo vivo que perciben los productores representa menos de la mitad del precio medio de exportación. Se concluye que la producción de equinos para carne requiere un plan nacional, lo cual daría como resultado mejores precios de venta para el productor y por ende un aumento en la producción.

Karyotypic relationships of horses and zebras: results of cross-species chromosome painting

Complete sets of chromosome-specific painting probes, derived from flow-sorted chromosomes of human (HSA), Equus caballus (ECA) and Equus burchelli (EBU) were used to delineate conserved chromosomal segments between human and Equits burchelli, and among four equid species, E. przewalskii (EPR), E. caballus, E. burchelli and E. zebra hartmannae (EZH) by cross-species chromosome painting. Genome-wide comparative maps between these species have been established. Twenty-two human autosomal probes revealed 48 conserved segments in E. burchelli. The adjacent segment combinations HSA3/21, 7/16p, 16q/19q, 14/15, 12/22 and 4/8, presumed ancestral syntenies for all eutherian mammals, were also found conserved in E. burchelli. The comparative maps of equids allow for the unequivocal characterization of chromosomal rearrangements that differentiate the karyotypes of these equid species. The karyotypes of E. przewalskii and E. caballus differ by one Robertsonian translocation (ECA5 = EPR23 + EPR24); numerous Robertsonian translocations and tandem fusions and several inversions account for the karyotypic differences between the horses and zebras. Our results shed new light on the karyotypic evolution of Equidae. Copyright (C) 2003 S. Karger AG, Basel.

Avaliação da taxa de fertilidade em éguas da raça puro sangue lusitano : efeito da idade da égua e do tipo de cobrição (cobrição natural vs inseminação artificial)

Ao longo dos meses de estágio acompanhei diversos procedimentos médicos veterinários, sendo que a maior parte deles diziam respeito à área da reprodução equina. A Raça Puro Sangue Lusitano (PSL) é a mais importante raça portuguesa autóctone de equinos. Contudo, não existem muitos estudos no que respeita à sua eficiência reprodutiva. Este trabalho pretende fazer um estudo sobre o efeito da idade da égua e do tipo de cobrição (natural e inseminação artificial - IA) na taxa de fertilidade de éguas PSL, através do cálculo das diferentes taxas de gestação. Neste estudo, verificou-se que a percentagem de éguas gestantes no final da época reprodutiva foi de 90%, encontrando-se dentro dos valores esperados (71% - 96%). Os dados foram recolhidos durante a época reprodutiva de 2011/2012 e permitiram-me elaborar um estudo retrospectivo para melhor perceber a prática do maneio reprodutivo equino e a influência de alguns parâmetros na taxa de fertilidade das éguas. Relativamente aos diferentes tipos de sémen, verificou-se que a taxa de fertilidade em éguas cobertas por cobrição natural (n=14) foi de 78,6% e por IA (n=36) foi de 94,4%, resultado que se situa dentro dos valores esperados. No que respeita à IA, a taxa de fertilidade com sémen fresco (n=16) foi de 93,8%, com sémen refrigerado (n=19) foi de 94,7% e com sémen congelado (n=1) foi de 100%. Os valores para sémen fresco e refrigerado encontram-se dentro do esperado, sendo que para sémen congelado este é muito superior aos valores reportados na bibliografia, visto ser o tipo de sémen com menores taxas de fertilidade. Este facto deve-se, provavelmente, à dimensão da amostra, e ao facto de o sémen congelado ser de elevada qualidade. Neste estudo verificamos também que não há relação estatística entre a idade das éguas e a taxa de gestação, contrariamente ao que é referido na bibliografia, o que se deverá provavelmente ao tamanho da amostra (n=50).

Relação dos centros de pressão dos quatro apoios do cavalo com a conformação dos respetivos membros

A conformação pode modular a predisposição para lesões de um cavalo. Por outro lado, o comportamento dos centros de pressão (CdP) dos cascos do cavalo pode fornecer informação relativa ao desempenho mecânico do seu sistema biológico. O objetivo deste trabalho foi testar a correlação dos CdP dos cascos com os principais defeitos de conformação da população estudada. Oito cavalos Puro Sangue Lusitano (PSL) saudáveis, com a cabeça numa posição neutra foram colocados, durante 15 ensaios de 8 segundos, com os membros anteriores (MA) numa placa de pressão e com os membros posteriores (MP) noutra. Os resultados sincronizados foram usados para caracterizar o deslocamento do CdP de cada casco e estudar a sua correlação com a conformação. Os resultados foram analisados de forma qualitativa, através da interpretação de áreas de contato e estabilogramas, assim como de forma quantitativa utilizando estatística inferencial para determinar uma eventual correlação com a conformação, recorrendo a um índice de conformação (IC). O defeito de conformação mais frequente foi os cavalos serem “esquerdos” e o que menos se observou foi serem “fechados de frente”. O IC variou entre 1 e 7. A média das áreas de contato dos cascos dos 8 cavalos foi maior nos MP do que nos MA, tendo acontecido o mesmo para os valores dos deslocamentos do CdP, que foram significativamente maiores nos MP do que nos MA Não foi detetada correlação entre os deslocamentos totais dos CdP e a conformação. Concluíu-se que o CdP dos PSL em estação não está dependente de variações de conformação em que o IC seja igual ou inferior a 7.

Viabilidade celular da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo de equinos após o processo de congelamento e descongelamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cardiorespiratory and endocrine effects of endogenous opioid antagonism by naloxone in ponies anaesthetised with halothane

Halothane depresses cardiorespiratory function and activates the pituitary-adrenal axis, increasing beta endorphin. In horses, beta endorphin may enhance the anaesthetic-associated cardiorespiratory depression and mortality risk. The authors studied endogenous opioid effects on cardiorespiratory function and pituitary-adrenal activity in halothane-anaesthetised ponies by investigating opioid antagonism by naloxone. Six ponies were anaesthetised three times (crossover design). Anaesthesia was induced with thiopentone and maintained with 1.2 per cent halothane for 2 hours. Immediately after induction, naloxone was administered either intra venously (0.5 mg kg(-1) bolus then 0.25 mg kg(-1) hour(-1) for 2 hours) or intrathecally (0.5 mg) or was replaced by saline as control. Pulse and respiratory rates, arterial blood gases, cardiac output and plasma cortisol and adrenocorticotrophic hormone (ACTH) concentrations were measured. All groups developed cardiorespiratory depression (40 per cent decrease in cardiac output) and plasma cortisol increased. Plasma ACTH concentration was higher in ponies treated with intrathecal naloxone. Endogenous opioids may inhibit ACTH Secretion, attenuating the stress response to halothane anaesthesia in equidae. (C) 2001 Harcourt Publishers Ltd.

Geologia e paleontologia de depósitos fossilíferos pleistocênicos do Rio Grande do Norte

This study deals with origin, sedimentary filling and fossil content of three tanks situated at Antônio Martins, Barcelona, Rui Barbosa, and Apodi counties, Rio Grande do Norte State. In addition, fossil materials from the Câmara Cascudo Museum - UFRN - Fundação Amigos do Lajedo Soledade - FALS, and from private ownership were investigated. The following families were identified: Megatberiidae, Gomphotheriidae, Mylodontidae, Equidae, Felidae, Canidae, Hydrochoeridae, Camelidae, Cervidae, Dasypodidae, Glyptodontidae, Macraucheniidae, Toxodontidae, and an undetermined Edentata Pilosa. The megafauna analysis indicated that herbivorc families occur mainly in tanks, whereas carnivore families occur in a vast proportion in an investigated ravine. Taphonomie analyses were limited to physical features because the vast majority of fossils were previously colleted without appropriate care for this kind of study. The main fossization processes were identified during diagenetie investigation. Permineralization is the most important process and replacement is the secondary one during fossilization. The study concluded that paleoenvironmental conditions during the late Pleistocene were more humid than the current one. Tropical savana, characterized by fields and cerrados , was the dominant vegetation

Characteristics of NOR-banded chromosomes of Mangalarga horses

The nucleolar organizer regions (NORs) of Mangalarga horses were characterized by analysis of NOR-banded metaphase chromosomes according to the technique of Goodpasture and Bloom (Chromosoma 53: 37-50, 1975). NOR banding was detected by silver staining in the telomeric region of the short arm of pair no. 1, in the region adjacent to the centromere of the long arm of pair no. 25 and in the proximal region of the long arm of pair no. 31. Associations of NOR-bearing chromosomal regions occurred in 12% of all metaphases and were frequent between the chromosomes of pair no. 1. Most (52.15%) of the NOR bands appeared on four chromosomes in both males and females. The maximum number of NOR-banded chromosomes was six, though only 11.34% of the cells examined showed this characteristic.