Determination of parent and substituted polycyclic aromatic hydrocarbons in high-fat salmon using a modified QuEChERS extraction, dispersive SPE and GC-MS

A fast and easy modified QuEChERS (quick, easy, cheap, rugged and safe) extraction method has been developed and validated for determination of 33 parent and substituted polycyclic aromatic hydrocarbons (PAHs) in high-fat smoked salmon that greatly enhances analyte recovery compared to traditional QuEChERS procedures. Sample processing includes extraction of PAHs into a solution of ethyl acetate, acetone and isooctane followed by cleanup with dispersive SPE and analysis by GC-MS in SIM mode. Method performance was assessed in spike recovery experiments (500 μg/g wet weight) in three commercially available smoked salmon with 3-11% fat. Recoveries of some 2-, 3- and 5-ring PAHs were improved 50-200% over traditional methods, while average recovery across all PAHs was improved 67%. Method precision was good with replicate extractions typically yielding relative standard deviations <10%, and detection limits were in the low ng/g range. With this method, a single analyst could extract and clean up ≥60 samples for PAH analysis in an 8 h work day.     

Novel 9-bp indel in visfatin gene and its associations with chicken growth

1. The effects of polymorphisms of the visfatin gene on growth performance, carcase traits, meat quality and serum variables were investigated in an F(2) resource population of Gushi chickens crossed with Anka broilers. 2. A 9-bp ('TAACCTGTG') insertion/deletion in intron10 of the visfatin gene was found and a total of 964 individuals were genotyped in the resource population. Genotypes (II, ID and DD) were identified based on the 9-bp insertion (allele I) or deletion (allele D). The insertion/deletion polymorphism was used for analysing associations of the gene with growth traits, carcase traits and meat quality traits in 414 F(2) chickens. 3. The DD genotype was not detected in those 66 F(1) chickens and the I allelic frequency (0·724-0·879) was obviously higher than the D allelic frequency (0·121-0·276) in the birds of three generations. 4. The 9-bp insertion/deletion was associated with the traits of 8-week shank length, 12-week shank length, 4-week pectoral angle and pancreas weight. The relationships with other traits: body weight, carcase traits, meat quality traits and serum variables, were not significant. 5. It was concluded that allele D (9-bp deletion) of the visfatin gene had a negative effect on skeletal growth.     

Use of multivariate analyses for determining heat tolerance in Brazilian cattle

Adaptability can be evaluated by the ability of an animal to adjust to environmental conditions and is especially important in extreme weather conditions such as that found in tropical Brazil. A multivariate analysis using physical and physiological traits in exotic (Nellore and Holstein) and naturalized (Junqueira, Curraleira, Mocho Nacional, Crioula Lageana, and Pantaneira) cattle breeds was carried out in the Federal District of Brazil to test and determine which traits are important in the adaptation of animal to heat stress as well as the ability of these traits and statistical techniques to separate the breeds studied. Both physical and physiological traits were measured on three occasions and included body measurements, skin and hair thickness, hair number and length, pigmentation, sweat gland area as well as heart and breathing rates, rectal temperature, sweating rate, and blood parameters. The data underwent multivariate statistical analyses, including cluster, discriminate, and canonical procedures. The tree diagram showed clear distances between the groups studied, and canonical analysis was able to separate individuals in groups. Coat traits explained little variation in physiological parameters. The traits which had higher discriminatory power included packed cell volume, shoulder height, mean corpuscular volume, body length, and heart girth. Morphological and physiological traits were able to discriminate between the breeds tested, with blood and size traits being the most important. More than 80% of animals of all breeds were correctly classified in their genetic group.     

Diagnosis of Eimeria species using traditional and molecular methods in field studies

The objective of this study was to identify and characterize species of Eimeria in broiler chickens using traditional morphological and pathological plus molecular (DNA amplification) diagnostic methodologies. Using a combination of those techniques it was possible to identify the presence of multiple circulating species in the flock as well as higher frequencies for some of them, especially Eimeria praecox and Eimeria maxima, which were identified in 100% of the flocks. The frequencies of the other species were Eimeria mitis and Eimeria necatrix (93.3%), Eimeria tenella (76,7%), Eimeria acervulina (56.7%) and Eimeria brunetti (16.7%). However using the lesion score, the most common species were E. maxima (46.7%), E. acervulina (30%), E. tenella (23.3%), and E. necatrix (10%). E. brunetti and E. praecox were not identified by using lesion score. DNA amplification had detection sensitivity for Eimeria species in the field samples of at least 20 oocysts. The implementation of DNA amplification as a routine diagnostic technique in aviaries can assist Eimeria population.     

A tumor-related lymphoid progenitor population supports hierarchical tumor organization in canine B-cell lymphoma

Background: Tumors have heterogeneous properties, which could be explained by the existence of hierarchically and biologically distinct tumor cells such as tumor‐initiating cells (TICs). This model is clinically important, as TICs are promising targets for cancer therapies. However, TICs in spontaneous B‐cell lymphoma have not been conclusively identified. Hypothesis/Objectives: Tumor cells with a progenitor phenotype exist in B‐cell lymphoma, reflecting a hierarchical organization. Animals: Twenty‐eight client‐owned dogs with previously untreated B‐cell lymphoma and 6 healthy dogs. Methods: This was a prospective study. Flow cytometry was used to identify lymphoid progenitor cells (LPCs) that coexpressed hematopoietic progenitor antigens CD34, CD117, and CD133, with lymphoid differentiation markers CD21 and/or CD22 in B‐cell lymphoma. The polymerase chain reaction for antigen receptor rearrangements was used to analyze clonality and relatedness of tumor populations. A xenograft model with NOD/SCID/IL‐2Rγ^?/? mice was adapted to expand and serially transplant primary canine B‐cell lymphoma. Results: LPCs were expanded in lymph nodes from 28 dogs with B‐cell lymphoma compared with 6 healthy dogs (P= .0022). LPCs contained a clonal antigen receptor gene rearrangement identical to that of the bulk of tumor cells. Canine B‐cell lymphoma xenografts in recipient mice that maintained LPCs in the tumors were recurrently observed. Conclusions and Clinical Importance: These results suggest the presence of a hierarchy of tumor cells in B‐cell lymphoma as has been demonstrated in other cancers. These findings have the potential to impact not only the understanding of lymphoma pathogenesis but also the development of lymphoma therapies by providing novel targets for therapy.     

Inoculants for ensiling low-dry matter corn crop: a midlactation cow perspective

In many regions, optimum dry matter (DM) content of corn crop pre-ensilage cannot be ensured for management, agronomical and climatic reasons. Under such conditions, corn crops are harvested at low DM, and are easily exposed to unfavourable fermentation pathways and plant spoilage and wastage. Thus, it is a major question for dairy agriculturists whether certain microbial inoculants application to low-DM corn crop pre-ensilage affects silage quality and cow performance. The objective was to determine effects of adding microbial inoculants to low-DM corn crop at ensiling on silage quality, rumen fermentation and milk production of eight Holstein cows fed the treated silages. Whole corn plant was harvested at milk stage of maturity with 204 g DM/kg of fresh crop, cut to a theoretical particle length of 2 cm, filled in 60 t bunker silos, and treated layer by layer with either no inoculant (control), inoculant 'E' (100 000 cfu/g of fresh crop) containing mainly Lactobacillus plantarum, inoculant 'B' (100 000 cfu) containing mainly Pediococcus pentosanus, Lactobacillus plantarum and Propionibacter freudenreichii or a mixture of inoculants 'E' and 'B' (200,000 cfu). Inoculants were mixed with water and sprayed on thin layers of corn chops layer by layer followed by rolling to ensure proper oxygen outage and even microbial distribution throughout the plants. Eight multiparous lactating Holstein cows at 100 ± 20.5 days in milk were used in a replicated 4 × 4 Latin square design with four 20-day periods including 14 days of adaptation and 6 days of sampling. Dietary treatments were mixed rations containing corn silages with or without the inoculants. The basal diet contained 32.9% corn silage, 14.3% alfalfa hay and 52.8% concentrate on a DM basis. Inoculants did not affect silage pH or content of DM, CP, lactate, acetate, ash and total volatile fatty acids (VFA). Applying 'B' to corn crop resulted in higher water soluble carbohydrates (47.7 g/kg vs 29.8 g/kg) and lower neutral detergent fibre (494.1 g/kg vs 464.0 g/kg) compared with control. The combined inoculants increased silage butyrate relative to other treatments. The mixture of 'E + B' and 'B' moderately decreased rumen pH, when compared to 'E'. The 'E + B' increased rumen VFA concentrations relative to 'E' and control silage. Dry matter intake increased when corn crop was ensiled with 'E' than with control and 'E + B', but this had little impact on milk production or its energy concentrations. Milk energy yield tended to decrease when 'B' but not 'E' was applied alone, compared with control and 'E + B'. The estimated proportion of the consumed energy secreted in milk increased when inoculants were applied together compared with when they were used separately. Results suggest positive effects of Lactobacillus plantarum containing inoculant on feed intake, some effects on corn silage water soluble carbohydrates, fibre and butyrate contents, rumen pH and VFA concentrations; but no significant effects on total tract nutrient digestibility or productivity of Holstein cows fed diets with 329 g corn silage/kg of diet DM.     

Aspects of meat and eating quality of broiler chickens reared under standard, maize-fed, free-range or organic systems

1. The effects of rearing chickens using standard, maize-fed, free-range or organic production systems on meat quality and sensory characteristics were evaluated. The standard system used either Ross or Cobb birds with a slaughter age of approximately 40 d, the other systems in the trial used Hubbard birds with considerably older slaughter ages, up to 72 d in the case of organic systems. 2. Paired breast fillets from 120 birds, 30 from each rearing system, were used. The meat quality variables, ultimate pH, colour coordinates L*, a*, b* and water-holding capacity were measured and taste panel assessments were made using 8-point category scales of texture, juiciness, abnormal flavour, flavour liking and overall flavour. 3. There were significant differences between rearing systems, with fillet muscles from birds grown under the standard system having a higher ultimate pH. Differences were also seen in colour with fillets from birds reared under a standard system having a smaller hue angle than those grown using the maize-fed system which had the highest. 4. Fillets from birds reared in the standard system were rated by the taste panel as more tender and juicy. There were no significant differences in chicken flavour. Based on hedonic assessments of flavour liking and overall liking, by a small panel of assessors, meat from birds produced in the standard system was most preferred and that from organic systems the least preferred. Meat from free-range and maize-fed systems was intermediate in preference. This result reveals a trend, but does not infer consumer acceptance.     

Neuropathology of italian cats in feline spongiform encephalopathy surveillance

Feline spongiform encephalopathy (FSE) is a transmissible spongiform encephalopathy associated with the consumption of feedstuffs contaminated with tissue from bovine spongiform encephalopathy-affected cattle and characterized by the accumulation in the central nervous system of an abnormal isoform of the prion protein (PrP(sc)). Clinically, it presents as a progressive fatal neurologic syndrome that is not easily distinguished from other feline neurologic conditions. Most cases of FSE have been reported in England, where it was first detected in 1990, but a few cases have been reported from other European countries. To identify possible cases of FSE in Italy, the Italian Ministry of Health funded a 2-year surveillance project during which the brains from 110 domestic cats with neurologic signs were evaluated histologically for spongiform encephalopathy and immunohistochemically to detect PrP(sc). Although no cases of FSE were found, the study proved useful in monitoring the Italian cat population for other neurologic diseases: neoplasia (21.8%), toxic-metabolic encephalopathy (18.2%), granulomatous encephalitis (15.5%), suppurative encephalitis (4.6%), trauma (3.6%), circulatory disorders (3.6%), degeneration (2.7%), nonsuppurative encephalitis (2.7%), and neuromuscular diseases (1.8%). No histologic lesions were found in 20% of the brains, and samples from 5.5% of the cats were rejected as unsuitable.