LOW-FIELD MRI AND ARTHROSCOPY OF MENISCAL LESIONS IN TEN DOGS WITH EXPERIMENTALLY INDUCED CRANIAL CRUCIATE LIGAMENT INSUFFICIENCY

Little is known about the magnetic resonance imaging (MRI) appearance of canine meniscal lesions. The aim of this study is to describe the MR appearance of meniscal lesions in dogs with experimentally induced cranial cruciate ligament (CCL) deficiency. The pilot study revealed dogs weighing approximately 10 kg to be too small for meniscal evaluation on low-field MRI. In the main study, dogs weighing approximately 35 kg were used. The left CCL was transected and low-field MRI was performed regularly until 13 months post-surgery. Normal menisci were defined as grade 0. Intrameniscal lesions not reaching any surface corresponded to grade 1 if focal and to grade 2 if linear or diffuse. Grade 3 lesions consisted in linear tears penetrating a meniscal surface. Grade 4 lesions included complex signal changes or meniscal distortion. Between 2 and 13 months post-surgery, all dogs developed grade 4 lesions in the medial meniscus. Most of them corresponded to longitudinal or bucket handle tears on arthroscopy and necropsy. Two dogs showed grade 3 lesions reaching the tibial surface of the lateral meniscus on MRI but not in arthroscopy. Such tears are difficult to evaluate arthroscopically; MRI provides more accurate information about the tibial meniscal surface. Grades 1 and 2 lesions could not be differentiated from presumably normal menisci with our imaging technique. An MRI grading system better adapted to canine lesions has yet to be developed. MRI is a helpful tool for the diagnosis of complete tears in the canine meniscus, especially in larger dogs.     

Comparison of methods to reduce dioxin and polychlorinated biphenyls contents in fishmeal: extraction and enzymatic treatments

Methods to remove dioxins and polychlorinated biphenyls (PCBs) from fishmeal were investigated and compared. The tested methods include (i) lowering the fat content and simultaneously the level of toxic contaminants using either organic solvents or (ii) lowering the fat content using protease and (iii) removal of dioxins and PCBs using either oil extraction or (iv) breakdown of dioxin and PCBs using oxidoreductase. The results showed that the organic solvents tested (ethanol, isopropanol, and isohexane) were efficiently lowering the oil content of the fishmeal by 80%. However, the treated fishmeal has a low fat content and may contain traces of solvent. The protease alcalase was not as efficient as the solvent extraction and only removed approximately 30% of the oil but presented the advantage of being a mild process. Other proteases, alone or in combination with other enzymes, might give better yield if the reaction conditions are optimized. In contrast, extraction of dioxin and PCBs using olive oil or fish oil was very effective and resulted in 60-75% extraction of dioxin and PCBs, respectively, after a single extraction step. No preference for the oil type was observed. This method is very simple and quick and does not require an important investment for the fishmeal producer. It is expected that with optimization this method could be implemented at an industrial scale without too many difficulties. In contrast, the oxidoreductases tested did not result in a major degradation of dioxins and PCBs with only 10-15% degradation achieved. However, with the recent advancement in biotechnology, it is possible that future research will result in the development of enzymes that effectively degrade recalcitrant contaminants.     

Oxidation in fish oil enriched mayonnaise: ascorbic acid and low pH increase oxidative deterioration

The effect of ascorbic acid (0-4000 ppm) and pH (3.8-6.2) on oxidation and levels of iron and copper in various fractions of mayonnaise enriched with 16% fish oil was investigated. Ascorbic acid induced release of iron from the assumed oil-water interface into the aqueous phase at all pH levels, but this effect of ascorbic acid was strongest at low pH (pH 3.8-4.2). Ascorbic acid generally promoted formation of volatile oxidation compounds and reduced the peroxide value in mayonnaises. Peroxide values and total volatiles generally increased with decreasing pH values, suggesting that low pH promoted oxidation. It is proposed that iron bridges between the egg yolk proteins low-density lipoproteins, lipovitellin, and phosvitin at the oil-water interface are broken at low pH values, whereby iron ions become accessible as oxidation initiators. In the presence of ascorbic acid, oxidation is further enhanced due to the reduction of Fe(3+) to Fe(2+) that rapidly catalyzes lipid oxidation via lipid hydroperoxide decomposition at the oil-water interface in mayonnaise.     

Partitioning of selected antioxidants in mayonnaise.

This study examined partitioning of alpha-, beta-, and gamma-tocopherol and six polar antioxidants (Trolox, ferulic acid, caffeic acid, propyl gallate, gallic acid, and catechin) in mayonnaise. Partitioning of antioxidants between different phases was determined after separation of mayonnaise by either (a) centrifugation + ultracentrifugation or (b) centrifugation + dialysis. Antioxidants partitioned in accordance with their chemical structure and polarity: Tocopherols were concentrated in the oil phase (93-96%), while the proportion of polar antioxidants in the oil phase ranged from 0% (gallic acid and catechin) to 83% (Trolox). Accordingly, proportions of 6% (Trolox) to 80% (gallic acid and catechin) were found in the aqueous phase. Similar trends were observed after dialysis. After ultracentrifugation, large proportions of polar antioxidants were found in the "emulsion phase" and the "precipitate" (7-34% and 2-7%, respectively). This indicated entrapment of antioxidants at the oil-water interface in mayonnaise. The results signify that antioxidants partitioning into different phases of real food emulsions may vary widely.     

Irradiation of diets fed to captive exotic felids: microbial destruction, consumption, and fecal consistency.

Two frozen, raw horse meat-based diets fed to captive exotic felids at Brookfield Zoo were irradiated to determine the extent of microbial destruction and whether radiation treatment would affect consumption and/or fecal consistency in exotic cats. Fifteen cats, two African lions (Panthera leo), two Amur tigers (Panthera tigris altaica), one Amur leopard (Panthera pardus orientalis), two clouded leopards (Neofelis nebulosa), two caracals (Felis caracal), one bobcat (Felis rufus), and five fishing cats (Felis viverrinus), housed at Brookfield Zoo were fed nonirradiated and irradiated raw diets containing horse meat with cereal products and fortified with nutrients: Nebraska Brand Feline and/or Canine Diet (Animal Spectrum, North Platte, Nebraska 69103, USA). Baseline data were obtained during a 2-wk control period (nonirradiated diets), which was followed by a 4-wk period of feeding comparable irradiated diets. Feed intake and fecal consistency data were collected. An estimated radiation dose range of 0.5-3.9 kilograys reduced most microbial populations, depending on specific diet and microbe type. Irradiation had no overall effect on either feed consumption or fecal consistency in captive exotic cats, regardless of species, age, sex, or body mass. Data indicate that irradiation of frozen horse meat-based diets (packaged in 2.2-kg portions) result in microbial destruction in these products but that product storage time between irradiation and sampling may also affect microbial reduction. However, irradiation would be an appropriate method for reducing potentially pathologic bacteria in raw meat fed to exotic cats.     

Requirement of CHROMOMETHYLASE3 for maintenance of CpXpG methylation

Epigenetic silenced alleles of the Arabidopsis SUPERMAN locus (the clark kent alleles) are associated with dense hypermethylation at noncanonical cytosines (CpXpG and asymmetric sites, where X = A, T, C, or G). A genetic screen for suppressors of a hypermethylated clark kent mutant identified nine loss-of-function alleles of CHROMOMETHYLASE3 (CMT3), a novel cytosine methyltransferase homolog. These cmt3 mutants display a wild-type morphology but exhibit decreased CpXpG methylation of the SUP gene and of other sequences throughout the genome. They also show reactivated expression of endogenous retrotransposon sequences. These results show that a non-CpG DNA methyltransferase is responsible for maintaining epigenetic gene silencing.     

Agronomical and nutritional evaluation of quinoa seeds (Chenopodium quinoa Willd.) as an ingredient in bread formulations

Quinoa is an Andean seed crop of many potential uses. In 2009 a field trial was carried out to explore the potential for quinoa growing in climatic conditions of South Eastern Europe. Even under rainfed conditions, without fertilization, a seed yield as high as 1.721 t ha⁻¹ was obtained. Seed quality was remarkably good, with protein content ranging from 15.16 to 17.41 % on a dry weight basis, depending on whether seeds were processed. Amino acid and mineral composition revealed the potential of quinoa seeds as a valuable ingredient in the preparation of highly nutritious foods. Quinoa seeds had higher contents of most essential amino acids, especially lysine, than wheat flour. Dehulled quinoa seeds, devoid of saponins, were included into wheat bread formulations, with up to 20%, which resulted in a positive effect on the rheological characteristics of dough. Furthermore, protein content in bread was increased by around 2%. Sensory characteristics of breads were excellent also at the 20% supplementation level. The study of bread supplemented with quinoa seeds could enable the development of a range of new baking products with enhanced nutritional value.     

Cisgenesis,a New Tool for Traditional Plant Breeding,Should be Exempted from the Regulation on Genetically Modified Organisms in a Step by Step Approach

Modern potato breeding requires over 100,000 seedlings per new variety. Main reasons are (1) the increasing number of traits that have to be combined in this tetraploid vegetatively propagated crop, and (2) an increasing number of traits (e.g., resistance to biotic stress) originates from wild species. Pre-breeding by introgression or induced translocation is an expensive way of transferring single traits (such as R-genes, coding for resistance to biotic stress) to the cultivated plant. The most important obstacle is simultaneous transfer of undesired neighbouring alien alleles as linkage drag. Stacking several genes from different wild sources is increasing this linkage drag problem tremendously. Biotechnology has enabled transformation of alien genes into the plant. Initially, transgenes were originating mainly from microorganisms, viruses or non-crossable plant species, or they were chimeric. Moreover, selection markers coding for antibiotic resistance or herbicide resistance were needed. Transgenes are a new gene source for plant breeding and, therefore, additional regulations like the EU Directive 2001/18/EC were developed. Because of a strong opposition against genetic modification of plants in Europe, the application of this Directive is strict, very expensive, hampering the introduction of genetically modified (GM) crops and the use of this technology by small and medium-sized enterprises (SMEs). Currently, GM crops are almost the exclusive domain of multinationals. Meanwhile, not only transgenes but also natural genes from the plant species itself or from crossable plant species, called cisgenes, are available and the alien selection genes can be avoided in the end product. This opens the way for cisgenic crops without alien genes. The existing EU directive for GM organisms is not designed for this new development. The cisgenes belong to the existing breeders’ gene pool. The use of this classical gene pool has been regulated already in agreements regarding breeders’ rights. We are proposing a step by step approach starting with a crop and gene specific derogation and monitoring towards a general exemption of cisgenic plants from the Directive. Two examples, i.e. development of cisgenic potato for resistance to Phytophthora infestans and cisgenic apple for resistance to Venturia inaequalis are discussed shortly for illustration of the importance of cisgenesis as a new tool for traditional plant breeding. Cisgenesis is simplifying introgression and induced translocation breeding tremendously and is highly recommended for SMEs and developing countries.