Implications and challenges of tuberculosis in wildlife ungulates in Portugal: a molecular epidemiology perspective

Mycobacterium bovis and, more rarely, Mycobacterium caprae, may cause zoonotic bovine tuberculosis (bTB) in an extensive range of animal species. In Portugal, during 2009, a remarkable raise of bTB incidence was registered in cattle along with an increase of new cases in wildlife. In this work, we reassess and update the molecular epidemiology of bTB in wild ungulates by including 83 new M. bovis and M. caprae isolates from wild boar and red deer obtained during 2008-2009. Spoligotyping identified 27 patterns in wild ungulates, including 11 patterns exclusive from deer and five from wild boar. The genetic relatedness of wildlife and livestock isolates is confirmed. However, the relative prevalence of the predominant genotypes is different between the two groups. Contrasting with the disease in livestock, which is widespread in the territory, the isolation of bTB in wildlife is, apparently, geographically localized and genotypic similarities of strains are observed at the Iberian level.     

Vaccine immune response and interference of colostral antibodies in calves vaccinated against rabies at 2, 4 and 6 months of age born from antirabies revaccinated females

Considering the high prevalence of rabies in cattle, we aimed to evaluate the interference of colostral antibodies transferred to calves after birth and the benefit of administering an antirabies vaccination in two-month-old calves compared to vaccinating at 4 and 6 months of age. Calves born from females revaccinated against rabies during the third trimester of pregnancy were studied. Forty-eight hours after parturition, blood samples from dams and offspring were collected, and antirabies neutralizing antibody titers were analyzed using the Rapid Focus Fluorescent Inhibition Test. We found that all calves had similar titers of antibodies transferred through the colostrum. Furthermore, none of the calves presented a satisfactory serological response after the first vaccination, but all had an appropriate response after revaccination. This study demonstrates that antirabies vaccination should be recommended for calves at two months of age in endemic and epizootic situations.     

Voluntary intake and milk production in F1 Holstein × zebu cows in confinement

The objective of this study was to evaluate the nutrient intake and milk production in Holstein × zebu (F1) cows in feedlot. Eighteen F1 cows were used, divided into three treatments; six were Holstein × Gir (HGI), six were Holstein × Guzerat (HGU), and another six were Holstein × Nelore (HNE), which had recently calved, distributed into simple, random samples, under the same feeding conditions of corn silage and concentrate with 20% crude protein. The three-marker method was used (LIPE, titanium dioxide, and iADF) to estimate the individual intake and digestibility of the nutrients for the cows in group. The mathematical model used to establish the lactation curves was: Y = at(b)e(-ct) by Wood (Nature 216:164-165, 1967). The statistical analyses for the nutrient intake and digestibility, as well as parameters of metabolic efficiency, were performed using multiple linear regression (α = 5%). No effect (P > 0.05) of genetic group was observed for any of the variables studied. The intake and digestibility of the nutrients and the microbial nitrogen presented quadratic curves as a function of the lactation period (P < 0.05). The HGU cows exhibited an accumulated milk production of 4,946.81 kg at 305 days, whereas the HGI cows produced 4,821.78 kg. The HNE cows displayed inferior performance, with a production of 3,674.98 kg. It was concluded that, in confinement, F1 cows from different genetic groups do not exhibit different intake, digestibility, or metabolic efficiency. The HGU and HGI cows have greater cumulative production at 305 days.     

Successful use of a gonadotropin-releasing hormone (GnRH) analog for the treatment of tertiary hypogonadism (GnRH deficiency) in a 5-year-old Belgian Blue bull

A bull was referred for a progressive oligoasthenotheratozoospermia that resulted in a unsuitable seminal quality for the cryopreservation. Breeding soundness evaluation results suggested gonadal dysfunction. Because of the lack of normal ranges for these hormones in the bull, in this study, the hypogonadism and the site of the dysfunction (hypothalamus) were diagnosed by the gonadotropin-releasing hormone (GnRH) stimulation test. The evaluation of pituitary and testicular responsiveness by a GnRH stimulating test revealed a responsiveness of the pituitary and testis, thus a secondary hypogonadism (hypothalamic hypogonadism) was postulated and a therapeutic approach based on the subcutaneous administration of GnRH analog was attempted. An increase in semen volume, concentration and sperm characteristics were detected 9 weeks after the start of the treatment, corroborating the hypothalamic origin of the disease and the useful of the GnRH therapy.     

Perspectives in dryland restoration: approaches for climate change adaptation

Reforestation efforts in dryland ecosystems frequently encounter drought and limited soil productivity, although both factors usually interact synergistically to worsen water stress for outplanted seedlings. Land degradation in drylands (e.g. desertification) usually reduces soil productivity and, especially, soil water availability. In dry sub-humid regions, forest fires constitute a major disturbance affecting ecosystem dynamics and reforestation planning. Climate change projections indicate an increase of drought and more severe fire regime in many dryland regions of the world. In this context, the main target of plantation technology development is to overcome transplant shock and likely adverse periods, and in drylands this is mostly related to water limitations. In this paper, we discuss some selected steps that we consider critical for improving success in outplanting woody plants, both under current and projected climate change conditions including: (1) Plant species selection, (2) Improved nursery techniques, and (3) Improved planting techniques. The number of plant species used in reforestation is increasing rapidly, moving from a reduced set of well-known, easy-to-grow, widely used species, to a large variety of promising native species. Available technologies allow for reintroducing native plants and recovering critical ecosystem functions for many degraded drylands. However, climate change projections introduce large uncertainties about the sustainability of current reforestation practices. To cope with these uncertainties, adaptive restoration approaches are suggested, on the basis of improved plant quality, improved techniques for optimizing rain use efficiency in plantations, and exploring native plant species, including provenances and genotypes, for their resilience to fire and water use efficiency.     

Slaframine toxicosis in Brazilian horses causing excessive salivation

Ingestion of food contaminated with slaframine, an alkaloid produced by Rhizoctonia leguminicola, causes a mycotoxicosis, characterised by excessive salivation. Twenty‐eight horses demonstrated this clinical sign after the consumption of alfalfa hay which on inspection showed dark patches on many of the stems. The presence of slaframine (1.5 ppm) in this hay was confirmed by gas chromatography and mass spectroscopy. This is the first equine slaframine toxicosis case reported in Brazil.     

Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.     

Resolving the Parentage of the Apple Cultivar ‘Meran’

In the mid-1970s, a new apple variety named ‘Meran’ was discovered in South Tyrol (northern Italy), which harbours the largest continuous apple growing area in Europe. The cultivar was registered for varietal protection and patented in several countries, and was declared to be a cross of the varieties ‘Golden Delicious’ and ‘Morgenduft’ (synonym ‘Rome Beauty’). The parentage of ‘Meran’ has, however, been questioned, and the present study aimed to assess the descent of this cultivar by the combined use of molecular genetic and bioinformatic tools. Five accessions of ‘Meran’ were collected from three different European germplasm collections and analysed at 14 variable microsatellite DNA loci. Subsequently, computer software was used to allocate the most likely parent pair from a set of cultivars representative for the apple growing area of South Tyrol in 1975. The molecular genetic data clearly excluded ‘Morgenduft’ as a gene donor to ‘Meran’ and provided strong evidence that ‘Meran’ is a cross of the cultivars ‘Golden Delicious’ and ‘Jonathan’, confirming previous assumptions based on morphological traits of the tree and fruit.     

Digital Agenesia in Martina Franca Donkey Foal: A Case Report

A 4-day-old male Martina Franca donkey foal was evaluated for a forelimb alteration. Clinical examination and radiographs revealed the agenesia of the distal digit. Biochemical parameters were normal, and ultrasonographic evaluation did not identify any relievable organ alteration. Karyotype study revealed an abnormality on chromosome 1. The foal was discharged with a distal limb bandage in which a palmar splint was applied. A poor prognosis for the functionality of the limb was given. In endangered species, such as the Martina Franca donkey, the excessive inbreeding could result in an increase in genetic disorders. These findings shed new light on the possible pathogenesis of the digital dysgenesia. The study of the karyotype could be a useful approach to detect genetic alterations that could or could not be expressed in the animal, especially in endangered species in which a risk of an excessive inbreeding is considerable. These defects should be considered in the choice and selection of the breeders.