Determination of tumor necrosis factor-alpha responsiveness in piglets around weaning using an ex vivo whole blood stimulation assay

Ex vivo whole blood stimulation with endotoxin has proved to be a useful method for quantitative evaluations of the pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) response ability in humans. In the present study, a dose- and time-response study was carried out in order to develop an ex vivo whole blood stimulation assay for the quantification of TNF-alpha production in pigs. The TNF-alpha response was enhanced with increasing endotoxin stimulation dose. The maximum TNF-alpha response occurred after 2-8 h of stimulation. Subsequently, the assay was used to evaluate the TNF-alpha response in pigs (n=32) in relation to weaning. The TNF-alpha response was 332 pg ml(-1) (+/-59 pg) plasma 2 days before weaning and 127 pg ml(-1) (+/-23 pg) plasma 2 days after weaning, which was a significant reduction (p<0.001). Total and differential counts of leucocytes were the same before and after weaning. Thus the lower TNF-alpha response may be due to reduced monocyte responsiveness to the endotoxin, rather than caused by a reduction in monocyte numbers. The reduced TNF-alpha response in piglets after weaning may be a factor of importance to the increased disease susceptibility seen in piglets in this period.     

Zoonotic aspects of Mycobacterium bovis and Mycobacterium avium-intracellulare complex (MAC)

Pathogens that are transmitted between the environment, wildlife, livestock and humans represent major challenges for the protection of human and domestic animal health, the economic sustainability of agriculture, and the conservation of wildlife. Among such pathogens, the genus Mycobacterium is well represented by M. bovis, the etiological agent of bovine tuberculosis, M. avium ssp. paratuberculosis (Map) the etiological agent of Johne disease, M. avium ssp. avium (Maa) and in a few common cases by other emergent environmental mycobacteria. Epidemiologic surveys performed in Europe, North America and New Zealand have demonstrated the existence and importance of environmental and wildlife reservoirs of mycobacterial infections that limit the attempts of disease control programmes. The aim of this review is to examine the zoonotic aspects of mycobacteria transmitted from the environment and wildlife. This work is focused on the species of two main groups of mycobacteria classified as important pathogens for humans and animals: first, M. bovis, the causative agent of bovine tuberculosis, which belongs to the M. tuberculosis complex and has a broad host range including wildlife, captive wildlife, domestic livestock, non-human primates and humans; the second group examined, is the M. avium-intracellulare complex (MAC) which includes M. avium ssp. avium causing major health problems in AIDS patients and M. avium ssp. paratuberculosis the etiological agent of Johne disease in cattle and identified in patients with Crohn disease. MAC agents, in addition to a broad host range, are environmental mycobacteria found in numerous biotopes including the soil, water, aerosols, protozoa, deep litter and fresh tropical vegetation. This review examines the possible reservoirs of these pathogens in the environment and in wildlife, their role as sources of infection in humans and animals and their health impact on humans. The possibilities of control and management programmes for these mycobacterial infections are examined with regards to the importance of their natural reservoirs.     

Vitamin D status of wild Ricord's iguanas (Cyclura ricordii) and captive and wild rhinoceros iguanas (Cyclura cornuta cornuta) in the Dominican Republic.

Calcidiol (25-hydroxyvitamin D) values are reported for 22 wild Ricord's iguanas (Cyclura ricordii) and seven wild rhinoceros iguanas (Cyclura cornuta cornuta). Calcitriol (1,25-hydroxyvitamin D) values are reported for 12 wild Ricord's iguanas and seven wild rhinoceros iguanas. These animals were captured as part of a larger health assessment study being conducted on Ricord's iguanas in Isla Cabritos National Park, Dominican Republic. A total of 13 captive rhinoceros iguanas held outdoors at Parque Zoológico Nacional were also sampled for comparison. Mean concentrations of 25-hydroxyvitamin D were 554 nmol/L (222 ng/ml) with a range of 250-1,118 nmol/L (100-448 ng/ml) for wild Ricord's iguanas, 332 nmol/L (133 ng/ml) with a range of 260-369 nmol/L (104-148 ng/ml) for wild rhinoceros iguanas, and 317 nmol/L (127 ng/ml) with a range of 220-519 nmol/L (88-208 ng/ml) for captive rhinoceros iguanas. On the basis of these results, serum concentrations of at least 325 nmol/L (130 ng/ml) for 25-hydroxyvitamin D should be considered normal for healthy Ricord's and rhinoceros iguanas.     

Digestion and metabolism of low and high residual feed intake Nellore bulls

Understanding the reasons why animals of similar performances have different feed requirements is important to increase profits for cattle producers and to decrease the environmental footprint of beef cattle production. This study was carried out aiming to identify the associations between residual feed intake (RFI) and animal performance, nutrient digestibility, and blood metabolites related to energy balance of young Nellore bulls during the finishing period. Animals previously classified as low (n?=?13) and high RFI (n?=?12), with average initial body weight of 398 kg and age of 503 days were used. Cattle were fed a high energy diet and were slaughtered when rib fat thickness measured by ultrasound between the 12th and 13th ribs reached the minimum of 4 mm. A completely randomized design was adopted, being data analyzed with a mixed model that included the random effect of slaughter group, the fixed effect of RFI class, and linear effect of the covariate feedlot time. No differences were found (p?>?0.10) between RFI classes for performance, dry matter, and nutrients intake. However, dry (p?=?0.0911) and organic matter (p?=?0.0876) digestibility tended to be lower, and digestibility of neutral detergent fiber corrected for ash and protein (p?=?0.0017), and total digestible nutrients (p?=?0.0657) were lower for high RFI animals, indicating lesser capacity of food utilization. Difference between low and high RFI animals was also found for blood cortisol at the end of the trial (p?=?0.0044), having low RFI animals lower cortisol concentrations. Differences in the ability to digest food can affect the efficiency of transforming feed into meat by Nellore cattle.     

Cloning and expression of pigeon IFN-γ gene

This is the first paper describing the cloning of pigeon IFN-γ gene (PiIFN-γ) and the analysis of the in vitro expressed recombinant protein. The PiIFN-γ gene was identified by RT-PCR as a 498 bp, fragment coding for a precursor protein of 165 amino acids instead of 164 amino acids, as observed in the other avian species. The recombinant protein was expressed in vitro by an eukaryotic system and the biological properties of the cytokine were tested using a chicken macrophage cell line. The high degree of amino acid and nucleotide identity, shared with the ChIFN-γ, and the fact that the pigeon protein was functional on chicken cells, indicates a cross-reactivity between pigeon and chicken IFN-γ. The detection of the PiIFN-γ could represent an useful instrument in understanding the role played by this cytokine in immune response related to vaccinations and infectious diseases in the pigeon.     

NECROTIZING ENCEPHALITIS IN A YORKSHIRE TERRIER: CLINICAL, IMAGING, AND PATHOLOGIC FINDINGS

A 5-year-old, male Yorkshire Terrier had chronic and progressive neurologic signs compatible with lesions in the right brain stem and right forebrain. In magnetic resonance images of the brain there were multifocal lesions at different stages of evolution, consistent with an inflammatory disease. The lesions were located in the white matter of the cerebrum, in the diencephalon and mesencephalon. These lesions were hypointense in T1-weighted images and hyperintense in T2-weighted images with no mass effect and minimal enhancement with gadolinium. Necrotizing encephalitis was confirmed post mortem.     

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.     

Native and exotic seed dispersal by the stone marten (Martes foina): implications for the regeneration of a relict climactic forest in central Portugal

The stone marten (Martes foina) is a carnivorous mammal that often consumes fleshy fruits, thus potentially promoting seed dispersion. The present study was developed in Bussaco National Forest, central Portugal, and aimed to assess the potential role of the stone marten in dispersing native and exotic plants in different forest landscape types. Seeds from stone marten scats and fleshy fruits were collected monthly and were thereafter identified and sowed in a nursery, following a randomized experimental setup. Plant emergence was monitored fortnightly. Generalized linear models were used to test for differences in time and success of emergence between seeds from scats and fruits of 3 native species (Rubus ulmifolius, Arbutus unedo and Celtis australis) and 1 exotic plant species with invasive behavior (Prunus laurocerasus). Fruit consumption by the stone martens significantly increased and accelerated the germination of the native R. ulmifolius but had no effect on the other 2 native species or on the invasive species. This suggested that stone martens contribute to gene flow and forest regeneration by dispersing native plant seeds. However, although the germination was not enhanced in the invasive species, the preference of stone martens for these fruits may potentially contribute to the proliferation of P. laurocerasus. Our study represents a contribution to better understanding the fauna and flora interactions, enabling for a more conscious and effective decision‐making in forest management.     

Assessment of the long‐acting ivermectin formulation in sheep: Further insight into potential pharmacokinetic interactions

The aim of the current study was to evaluate the in vivo pharmacokinetic of ivermectin (IVM) after the administration of a long‐acting (LA) formulation to sheep and its impact on potential drug‐drug interactions. The work included the evaluation of the comparative plasma profiles of IVM administered at a single therapeutic dose (200 μg/kg) and as LA formulation at 630 μg/kg. Additionally, IVM was measured in different gastrointestinal tissues at 15 days posttreatment with both IVM formulations. The impact of the long‐lasting and enhanced IVM exposure on the disposition kinetics of abamectin (ABM) was also assessed. Plasma (IVM and ABM) and gastrointestinal (IVM) concentrations were analyzed by HPLC with fluorescent detection. In plasma, the calculated C_max and AUC_0‐t values of the IVM‐LA formulation were 1.47‐ and 3.35‐fold higher compared with IVM 1% formulation, respectively. The T_1/2ab and T_max collected after administration of the LA formulation were 2‐ and 3.5‐fold longer than those observed after administration of IVM 1% formulation, respectively. Significantly higher IVM concentrations were measured in the intestine mucosal tissues and luminal contents with the LA formulation, and in the liver, the increase was 7‐fold higher than conventional formulation. There was no drug interaction between IVM and ABM after the single administration of ABM at 15 days post‐administration of the IVM LA formulation. The characterization of the kinetic behavior of the LA formulation to sheep and its potential influence on drug‐drug interactions is a further contribution to the field.