Toxoplasma gondii: Detection by mouse bioassay, histopathology, and polymerase chain reaction in tissues from experimentally infected pigs

In the present study, we evaluated three techniques, mouse bioassay, histopathology, and polymerase chain reaction (PCR) to detect Toxoplasma gondii infection in tissues from experimentally infected pigs. Twelve mixed breed pigs, seronegative for T. gondii using an indirect immunofluorescent antibody test (IFAT), were used. Ten pigs were infected with 4 × 104 VEG strain oocysts, and two were maintained as uninfected controls. Animals were killed 60 days pos infection. Muscle (heart, tongue, diaphragm, and masseter) and brain samples were collected to investigate the presence of T. gondii tissue cysts by the different assay methods. For the bioassay, samples of brain (50 g) and pool of muscle samples (12.5 g of tongue, masseter, diaphragm, and heart) were used. PCR was performed using Tox4 and Tox5 primers which amplified a 529 bp fragment. The DNA extraction and PCR were performed three times, and all tissue samples were tested individually (brain, tongue, masseter, diaphragm, and heart). For histopathology, fragments of tissues were fixed in 10% of buffered formal saline and stained with HE. Histopathological results were all negative. PCR showed 25/150 (16.6%) positive samples, being 17/120 (14.1%) and 8/30 (26.6%) from muscle, and brain tissues, respectively. Tissue cysts of T. gondii were identified by mouse bioassay in 54/98 (55.1%) samples, being 31/48 (64.6%) from muscle samples, and 23/50 (46.0%) from brain samples. Toxoplasma gondii isolation in muscle samples by mouse bioassay was higher than in PCR (P < 0.01). Results indicate that DNA from pig tissues interfered with 529-bp-PCR sensitivity, and mouse bioassay was better than PCR in detecting T. gondii in tissues from pigs. © 2006 Elsevier Inc. All rights reserved.

Mananoligossacarídeo em dietas para leitões desmamados

An assay was carried out to evaluate the use of mannanoligosaccharide (MOS) in piglet diets on performance, diarrhea incidence and blood parameters. Different levels of MOS inclusion (0, 0.1 and 0.2%) for pig diets were compared. A total of 72 piglets of Topigs lineage weaned at 21 days of age with 5.28±0.90 kg of live weight were used. It was used a randomized block design to control differences between initial weights of replicates. The results show that MOS inclusion in weaning pig diets did not promote better results on daily weight gain, daily feed intake and feed conversion. Although reduction in diarrhea incidence was observed in animals fed with 0.2% MOS diet, this prebiotic did not improve the immune response of piglets. Any level of MOS evaluated is recommended for piglets.

Desvio da proporção sexual por meio da suplementação de fêmeas suínas com ácido ascórbico e sua influência na resposta à seleção

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização genética de javalis por meio de maracdores microssatélites

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Origem do suíno casco-de-burro e sua relação genética com populações ibéricas e americanas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Manure production and mineral excretion in feces of gilts fed ractopamine

A study was conducted to evaluate the feces+urine produced per animal (FUPA), dry matter, mineral matter, organic matter, nitrogen, phosphorus, potassium and sulfur in feces of gilts fed diets with increasing levels of ractopamine (0, 5, 10 and 15 mg kg-1 of diet). A total of 468 finishing gilts were allotted into 36 pens. In two days of each week, feces and urine were daily sampled in four pens per treatment, quantifying the feces+urine. To determine the characterization of feces, two samples per week were taken daily, in nine pens per treatment. It was used a split plot design, considering the ractopamine level as the plot and the weeks as the subplots. There was no reduction in nitrogen amount in feces. An interaction was detected between ractopamine concentrations and weeks for FUPA and phosphorus, potassium and sulfur in feces. Ractopamine addition in diets for gilts has reduced the feces+urine production and nitrogen and phosphorus excretion. Higher values estimated for potassium content in feces of animals fed diets with 10 and 15 mg of ractopamine kg-1 were found between the second and third week. Increasing levels of ractopamine from 5 to 15 mg kg-1 promoted higher excretion of sulfur over the weeks of supply.

Molecular detection of the blood meal source of sand flies (Diptera: Psychodidae) in a transmission area of American cutaneous leishmaniasis, Parana State, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Hypersensitivity induced in dogs by nymphal extract of Amblyomma cajennense ticks (Acari:Ixodidae)

In general, hosts develop resistance to ticks after repeated infestations; nevertheless, several studies on naturally occurring host-tick interactions were unable to detect resistance of hosts to ticks even after repeated infestations. The purpose of this investigation was to study the type of cutaneous hypersensitivity to unfed nymphal extract of A. cajennense in dogs, which, unlike guinea pigs, do not develop resistance. A first, but no second, peak in skin reaction was observed, suggesting that cellular immunity is an important mechanism of resistance to ticks. This may partially explain why guinea pigs, but not dogs, develop resistance against ticks.

Diet overlap and foraging activity between feral pigs and native peccaries in the pantanal

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

Detecção de anticorpos contra Leptospira spp. em animais de vida livre do Pantanal do Mato Grosso do Sul

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Wild Boar and Red Deer Display High Prevalences of Tuberculosis-Like Lesions in Spain

We describe the distribution of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red deer (Cervus elaphus) in Spain. Animals with TBL were confirmed in 84.21% of mixed populations (n = 57) of red deer and wild boar and in 75% of populations of wild boar alone (n = 8) in central and southern Spain (core area). The prevalence of TBL declined towards the periphery of this region. In the core area, the prevalence ranged up to 100% in local populations of wild boar (mean estate prevalence 42.51%) and up to 50% in red deer (mean estate prevalence 13.70%). We carried out exploratory statistical analyses to describe the epidemiology of TBL in both species throughout the core area. Prevalence of TBL increased with age in both species. Wild boar and red deer mean TBL prevalence at the estate level were positively associated, and lesion scores were consistently higher in wild boars than in red deer. The wild boar prevalence of TBL in wild boar did not differ between populations that were or were not cohabiting with red deer. Amongst the wild boars with TBL, 61.19% presented generalized lesions, and the proportion of generalized cases was similar between sex and age classes. In red deer, 57.14% of TBL-positive individuals presented generalized lesions, and the percentage of generalized cases increased with age class, but did not differ between the sexes. These results highlight the potential importance of wild boar and red deer in the maintenance of tuberculosis in south central Spain.

Risk Factors Associated With the Prevalence of Tuberculosis-Like Lesions in Fenced Wild Boar and Red Deer in South Central Spain

In recent decades the management of large game mammals has become increasingly intensive in south central Spain (SCS), resulting in complex epidemiological scenarios for disease maintenance, and has probably impeded schemes to eradicate tuberculosis (TB) in domestic livestock. We conducted an analysis of risk factors which investigated associations between the pattern of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red deer (Cervus elaphus) across 19 hunting estates from SCS and an extensive set of variables related to game management, land use and habitat structure. The aggregation of wild boar at artificial watering sites was significantly associated with an increasing risk of detecting TBL in both species, which probably relates to enhanced opportunities for transmission. Aggregation of wild boar at feeding sites was also associated with increased risks of TBL in red deer. Hardwood Quercus spp. forest availability was marginally associated with an increased risk of TB in both species, whereas scrubland cover was associated with a reduced individual risk of TBL in the wild boar. It is concluded that management practices that encourage the aggregation of hosts, and some characteristics of Mediterranean habitats could increase the frequency and probability of both direct and indirect transmission of TB. These findings are of concern for both veterinary and public health authorities, and reveal tuberculosis itself as a potential limiting factor for the development and sustainability of such intensive game management systems in Spanish Mediterranean habitats.

Feral swine damage and damage management in forested ecosystems

Invasive feral swine (Sus scrofa) cause deleterious impacts to ecosystem processes and functioning throughout their worldwide distribution, including forested ecosystems in the United States. Unfortunately, many feral swine damage management programs are conducted in a piecemeal fashion, are not adequately funded, and lack clearly stated or realistic objectives. This review paper identifies damage caused by feral swine to forest resources and presents techniques used to prevent and control feral swine damage. Concluding points related to planning a feral swine damage management program are: (1) the value of using a variety of techniques in an integrated fashion cannot be overstated; (2) there is value in using indices for both feral swine populations and their damage pre and post management activities; (3) innovative technologies will increasing be of value in the pursuit of feral swine damage reduction; and (4) though not appropriate in every situation, there is value in involving the public in feral swine damage management decisions and activities.

Development of spermatogenesis in captive-bred Spix's yellow-toothed cavy (Galea spixii)

The aim of this study was to evaluate the phases of sexual development and spermatogenesis of Spix's yellow-toothed cavy (Galea spixii) based on analyses of the structural components of the testes. The testes of animals from 0 to 150 days of age were collected by orchiectomy, weighed, and processed for analysis by light microscopy. At 45 days of age, spermatozoa were seen in the tubular lumen. Spermatogenesis was not established in animals from 45 to 150 days of age. The stages of sexual development may be classified into the following phases: from birth to the age of 15 days (immature); 30 days of age (prepubertal); 45-105 days of age (pubertal); and 120 and 150 days of age (postpubertal). This is the first study to address the male reproductive biology of Spix's yellow-toothed cavy.

Genetic Manipulation, Gene Silencing and Biomarker Development in Multiple Experimental Models.

The thesis is set in three different parts, according to the relative experimental models. First, the domestic pig (Sus scrofa) is part of the study on reproductive biotechnologies: the transgenesis technique of Sperm Mediated Gene Transfer is widely studied starting from the quality of the semen, through the study of multiple uptakes of exogenous DNA and lastly used in the production of multi-transgenic blastocysts. Finally we managed to couple the transgenesis pipeline with sperm sorting and therefore produced transgenic embryos of predetermined sex. In the second part of the thesis the attention is on the fruit fly (Drosophila melanogaster) and on its derived cell line: the S2 cells. The in vitro and in vivo models are used to develop and validate an efficient way to knock down the myc gene. First an efficient in vitro protocol is described, than we demonstrate how the decrease in myc transcript remarkably affects the ribosome biogenesis through the study of Polysome gradients, rRNA content and qPCR. In vivo we identified two optimal drivers for the conditional silencing of myc, once the flies are fed with RU486: the first one is throughout the whole body (Tubulin), while the second is a head fat body driver (S32). With these results we present a very efficient model to study the role of myc in multiple aspects of translation. In the third and last part, the focus is on human derived lung fibroblasts (hLF-1), mouse tail fibroblasts and mouse tissues. We developed an efficient assay to quantify the total protein content of the nucleus on a single cell level via fluorescence. We coupled the protocol with classical immunofluorescence so to have at the same time general and particular information, demonstrating that during senescence nuclear proteins increase by 1.8 fold either in human cells, mouse cells and mouse tissues.