Monoclonal antibodies specific to thermostable proteins in animal blood

Bovine plasma proteins are used as high-quality protein supplements in animal feed and as binders or colorants in food for human consumption. Religious observance, as well as recent fears of epidemic bovine spongiform encephalopathy, highlights the need for methods to detect bovine blood in processed food and animal feed for regulatory purposes, as the currently available methods are neither species-specific, blood-specific, nor valid for excessively heat-processed samples. This paper reports the development of monoclonal antibodies (MAbs) raised against bovine thermostable plasma proteins that display a unique species specificity pattern for plasma proteins. Immunoblotting revealed several thermostable antigenic proteins (10, 25, 40, and 60 kDa) in bovine plasma sterilized at 121 degrees C for 15 min. These MAbs can be employed individually or combined in immunoassays for analytical purposes and investigations of the chemical and biological properties of the thermostable plasma proteins identified here.     

Global genotype by environment trends in growth traits for Eucalyptus dunnii

New Forests - Eucalyptus dunnii Maiden (Dunn’s white gum) is a fast-growing tall tree with a restricted natural distribution in south-east Australia and favorable wood properties for pulp...     

PROGRESS ON IMPROVING AGRICULTURAL NITROGEN USE EFFICIENCY: UK-CHINA VIRTUAL JOINT CENTERS ON NITROGEN AGRONOM

● Virtual joint centers on N agronomy were established between UK and China. ● Key themes were improving NUE for fertilizers, utilizing livestock manures, and soil health. ● Improved management practices and technologies were identified and assessed. ● Fertilizer emissions and improved manure management are key targets for mitigation. Two virtual joint centers for nitrogen agronomy were established between the UK and China to facilitate collaborative research aimed at improving nitrogen use efficiency (NUE) in agricultural production systems and reducing losses of reactive N to the environment. Major focus areas were improving fertilizer NUE, use of livestock manures, soil health, and policy development and knowledge exchange. Improvements to fertilizer NUE included attention to application rate in the context of yield potential and economic considerations and the potential of improved practices including enhanced efficiency fertilizers, plastic film mulching and cropping design. Improved utilization of livestock manures requires knowledge of the available nutrient content, appropriate manure processing technologies and integrated nutrient management practices. Soil carbon, acidification and biodiversity were considered as important aspects of soil health. Both centers identified a range of potential actions that could be taken to improve N management, and the research conducted has highlighted the importance of developing a systems-level approach to assessing improvement in the overall efficiency of N management and avoiding unintended secondary effects from individual interventions. Within this context, the management of fertilizer emissions and livestock manure at the farm and regional scales appear to be particularly important targets for mitigation.     

Carbohydrate metabolism and hepatic zonation in the Atlantic hagfish,Myxine glutinosa liver: effects of hormones

Viable Atlantic hagfish (Myxine glutinosa) hepatocytes were isolated from combined or separated large and small lobes and carbohydrate metabolism was studied. Cells had low levels of glycogen (16–30 μmol·g⁻¹), and low rates of total glucose production (TGP; 0–480 nmol·h⁻¹·g⁻¹ cells). Lactate flux to glucose (5.5 nmol·h⁻¹·g⁻¹) and CO₂ (76 nmol·h⁻¹·g⁻¹) was lower than reported values for teleosts, with a low percentage (30%) of the lactate carbon reaching glucose. Insulin significantly increased total glucose production and gluconeogenesis and decreased 6-phosphofructo 1-kinase (PFK-1) activities and glucose oxidation, while glucagon was without effect on any parameter studied. Forskolin significantly increased TGP. Epinephrine (Epi), norepinephrine (NEpi), isoproterenol (Iso), and phenylephrine (Phe) all decreased CO₂ production from lactate; propanolol blocked the effects of Epi, NEpi, and Iso. The large lobe, accounting for 65% of total liver mass, had a higher glycogen content and higher CO₂ production from lactate compared to the small lobe. Furthermore, enzyme activities in the large lobe were greater than in the small lobe, with the exception of glycogen phosphorylase (GPase) which exhibited smaller %a values in the large lobe. These data indicate the presence of a hormonally-responsive carbohydrate metabolism in hagfish hepatocytes, which is qualitatively and quantitatively different between the two liver lobes.     

Comparison of fecal culture and F57 real-time polymerase chain reaction for the detection of Mycobacterium avium subspecies paratuberculosis in Swiss cattle herds with a history of paratuberculosis

Background

Bovine paratuberculosis is an incurable chronic granulomatous enteritis caused by Mycobacterium avium subspecies paratuberculosis (MAP). The prevalence of MAP in the Swiss cattle population is hard to estimate, since only a few cases of clinical paratuberculosis are reported to the Swiss Federal Food Safety and Veterinary Office each year.Fecal samples from 1,339 cattle (855 animals from 12 dairy herds, 484 animals from 11 suckling cow herds, all herds with a history of sporadic paratuberculosis) were investigated by culture and real-time polymerase chain reaction (PCR) for shedding of MAP.

Results

By culture, MAP was detected in 62 of 445 fecal pools (13.9%), whereas PCR detected MAP in 9 of 445 pools (2.0%). All 186 samples of the 62 culture-positive pools were reanalyzed individually. By culture, MAP was grown from 59 individual samples (31.7%), whereas PCR detected MAP in 12 individual samples (6.5%), all of which came from animals showing symptoms of paratuberculosis during the study. Overall, MAP was detected in 10 out of 12 dairy herds (83.3%) and in 8 out of 11 suckling cow herds (72.7%).

Conclusions

There is a serious clinically inapparent MAP reservoir in the Swiss cattle population. PCR cannot replace culture to identify individual MAP shedders but is suitable to identify MAP-infected herds, given that the amount of MAP shed in feces is increasing in diseased animals or in animals in the phase of transition to clinical disease.     

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

Zero-inflated models for identifying disease risk factors when case detection is imperfect: Application to highly pathogenic avian influenza H5N1 in Thailand

Logistic regression models integrating disease presence/absence data are widely used to identify risk factors for a given disease. However, when data arise from imperfect surveillance systems, the interpretation of results is confusing since explanatory variables can be related either to the occurrence of the disease or to the efficiency of the surveillance system. As an alternative, we present spatial and non-spatial zero-inflated Poisson (ZIP) regressions for modelling the number of highly pathogenic avian influenza (HPAI) H5N1 outbreaks that were reported at subdistrict level in Thailand during the second epidemic wave (July 3rd 2004 to May 5th 2005). The spatial ZIP model fitted the data more effectively than its non-spatial version. This model clarified the role of the different variables: for example, results suggested that human population density was not associated with the disease occurrence but was rather associated with the number of reported outbreaks given disease occurrence. In addition, these models allowed estimating that 902 (95% CI 881–922) subdistricts suffered at least one HPAI H5N1 outbreak in Thailand although only 779 were reported to veterinary authorities, leading to a general surveillance sensitivity of 86.4% (95% CI 84.5–88.4). Finally, the outputs of the spatial ZIP model revealed the spatial distribution of the probability that a subdistrict could have been a false negative. The methodology presented here can easily be adapted to other animal health contexts.     

Effects of dietary thyroid hormones on the red drum (Sciaenops ocellatus)

Four separate 8-week feeding trials were conducted to assess the effects of supplementing semipurified diets with either triiodothyronine (T₃) or thyroxine (T₄) at 0, 2, 10, and 50 mg/kg on growth and body composition of juvenile red drum (Sciaenops ocellatus) held in artificial brackish water (6‰) and artificial seawater (32‰). At both levels of salinity, increasing doses of T₃ resulted in fish with reduced weight gain, feed efficiency, condition factor (weight × 100/length³), and muscle ratio (muscle weight × 100/body weight), as well as a lighter body color. Significant (p < 0.05) effects of T₃ on the proximate composition of whole body, liver, and muscle were variable, generally reflecting decreased lipid and protein storage in liver and muscle, respectively. The two highest doses of T₃ given to seawater adapted fish increased survival. Dietary T₄ supplementation had no distinctive effects on appearance, growth or proximate body composition. These results indicate that whereas T₃ may function to regulate protein and lipid metabolism in red drum, dietary supplementation with T₃ leads to a hyperthyroidism-induced catabolic state. The elevated endogenous thyroid hormone levels found in fish fed optimal diets may thus adequately supply tissue needs during juvenile growth.     

Comparison of arylalkylamine N-acetyltransferase and melatonin receptor type 1B immunoreactivity between young adult and aged canine spinal cord

Melatonin affects diverse physiological functions through its receptor and plays an important role in the central nervous system. In the present study, we compared immunoreactivity patterns of arylalkylamine N-acetyltransferase (AANAT), an enzyme essential for melatonin synthesis, and melatonin receptor type 1B (MT2) in the spinal cord of young adult (2~3 years) and aged (10~12 years) beagle dogs using immunohistochemistry and Western blotting. AANAT-specific immunoreactivity was observed in the nuclei of spinal neurons, and was significantly increased in aged dog spinal neurons compared to young adult spinal neurons. MT2-specific immunoreactivity was found in the cytoplasm of spinal neurons, and was predominantly increased in the margin of the neuron cytoplasm in aged spinal cord compared to that in the young adult dogs. These increased levels of AANAT and MT2 immunoreactivity in aged spinal cord might be a feature of normal aging and associated with a feedback mechanism that compensates for decreased production of melatonin during aging.