Comparison of urine and blood as a convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium

Blood and urine were compared to clarify which is a better sample for estimating the concentration of radioactive cesium in muscles of live cattle. The most probable concentration of ¹³⁷Cs in muscles was 21.0 times that in blood, and the error of this estimation was 28%. The concentration of ¹³⁷Cs in blood was estimated accurately using urine samples. The estimation error did not increase so much (33%), even when the concentration of ¹³⁷Cs in muscles was estimated using urine samples. On the other hand, the maximum volume of blood that can be collected with one syringe is 50 ml, whereas it is easy to collect 1,000 ml of urine. It took at least 360 min to confirm that a cow meets the legal standard by inspecting 50 ml of blood. However, with 1,000 ml of urine, a 20 min measurement time was sufficient for this purpose. This difference in the required measurement time is critically important for practical use. In addition, urine can be collected by farmers themselves, whereas the blood collection requires a veterinarian. Therefore, urine is a more convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium.     

Distribution of radioactive cesium in edible parts of cattle

After the disastrous incident of the Fukushima Daiichi Nuclear Power Station, various agricultural, livestock and fishery products have been inspected for radioactive contamination with cesium in Japan. In this study, radioactive cesium was measured in various edible parts of cattle to verify the current inspection method for cattle, in which the neck tissues are generally used as samples. Radioactive cesium concentration in the short plate, diaphragm, liver, lung, omasum, abomasum and small intestine were lower and sirloin, tenderloin, top round meat and tongue were higher than that in the neck. There was no significant difference between the other organs (heart, kidney, lumen and reticulum) and the neck. Ninety‐five percent upper tolerance limits of the relative concentration to the neck were 1.88 for sirloin, 1.74 for tenderloin, 1.87 for top round and 1.45 for tongue. These results suggest that a safety factor of 2 is recommended for the radioactivity inspection of cattle to prevent a marketing of meat with higher cesium than the legal limit. Re‐inspection should be conducted using another part of muscle, for example, top round, when suspicious levels of 50–100 Bq/kg are detected in the neck.     

Software development for estimating the concentration of radioactive cesium in the skeletal muscles of cattle from blood samples

The 2011 earthquake severely damaged the Fukushima Daiichi Nuclear Power Plant (FNPP), resulting in the release of large quantities of radioactive material into the environment. The deposition of these radionuclides in rice straw as livestock feed led to the circulation of contaminated beef in the market. Based on the safety concern of the consumers, a reliable method for estimating concentrations of radioactive cesium in muscle tissue is needed. In this study, we analyzed the concentrations of radioactive cesium in the blood and skeletal muscle of 88 cattle, and detected a linear correlation between them. We then developed software that can be used to estimate radioactive cesium concentrations in muscle tissue from blood samples. Distribution of this software to the livestock production field would allow us to easily identify high‐risk cattle, which would be beyond the safety regulation, before shipping out to the market. This software is planned to be released as freeware. This software would contribute to food safety, and aid the recovery of the livestock industry from the damage creacted by the 2011 Tohoku earthquake and tsunami.     

Distribution of radioactive cesium and stable cesium in cattle kept on a highly contaminated area of Fukushima nuclear accident

Radioactivity inspection of slaughtered cattle is generally conducted using a portion of the neck muscle; however, there is limited information about the distribution of radioactive cesium in cattle. In this study, therefore, we measured not only radioactive cesium but also stable cesium in various tissues of 19 cattle that had been kept in the area highly contaminated by the Fukushima nuclear accident. Skeletal muscles showed approximately 1.5‐3.0 times higher concentration of radioactive cesium than internal organs. Radioactive cesium concentration in the tenderloin and top round was about 1.2 times as high as that in the neck muscle. The kidney showed the highest concentration of radioactive cesium among internal organs, whereas the liver was lowest. Radioactive cesium concentration in the blood was about 8% of that in the neck muscle. Characteristics of stable cesium distribution were almost the same as those of radioactive cesium. Correlation coefficient between radioactive cesium and stable cesium in tissues of individual cattle was 0.981 ± 0.012. When a suspicious level near 100 Bq/kg is detected in the neck of slaughtered cattle, re‐inspection should be conducted using a different region of muscle, for example top round, to prevent marketing of beef that violates the Food Sanitation Act.     

Radioactive cesium and potassium in cattle living in the ‘zone in preparation for the lifting of the evacuation order’ of the Fukushima nuclear accident

To contribute to the reconstruction of livestock industry in Fukushima, radioactive cesium (¹³⁴Cs, ¹³⁷Cs) and potassium (⁴°K) were measured in various tissues of beef cattle living in an area where the evacuation order will be lifted in the near future. Radioactive cesium concentration was less than 100 Bq/kg in most of the samples. Skeletal muscles and kidney had the highest concentrations of radioactive cesium, whereas the liver was lowest among samples, excepting blood. Radioactive cesium concentration in the sirloin, tenderloin and top round was significantly higher than that in the neck muscle. Radioactive cesium concentration in the urine was not correlated with that in the blood, but the relationship became proportional when corrected with urinary ⁴°K. Distribution of ⁴°K was similar but not identical to that of radioactive cesium. These results suggest that it will be possible to resume livestock production in this area after the decontamination measures are completed and the evacuation order is lifted. Contamination level of living cattle can be estimated not only by blood samples but also by urine samples. If 50–100 Bq/kg of cesium is detected in the neck muscle at a slaughterhouse, the cattle should be reinspected using a sample from different muscles.     

Cesium radioactivity in peripheral blood is linearly correlated to that in skeletal muscle: Analyses of cattle within the evacuation zone of the Fukushima Daiichi Nuclear Power Plant

The accident at the Fukushima Daiichi Nuclear Power Plant (FNPP) released a large amount of radioactive substances into the environment. Furthermore, beef contaminated with radioactive cesium above the 500 Bq/kg safety standard was circulated in the food chain in 2011. Japanese consumers remain concerned about the safety of radioactively contaminated food. In our previous study, we detected a linear correlation between radioactive cesium (¹³⁷Cs) activity in blood and muscle around 500 to 2500 Bq/kg in cattle. However, it was unclear whether the correlation was maintained at a lower radioactivity close to the current safety standard of 100 Bq/kg. In this study, we evaluated 17 cattle in the FNPP evacuation zone that had a ¹³⁷Cs blood level less than 10 Bq/kg. The results showed a linear correlation between blood ¹³⁷Cs and muscle ¹³⁷Cs (Y = 28.0X, R² = 0.590) at low radioactivity concentration, indicating that cesium radioactivity in the muscle can be estimated from blood radioactivity. This technique would be useful in detecting high‐risk cattle before they enter the market, and will contribute to food safety.     

Decreased blood cell counts were not observed in cattle living in the “difficult‐to‐return zone” of the Fukushima nuclear accident

White blood cells, especially lymphocytes, are susceptible to radiation exposure. In the present study, red blood cell, total white blood cell, and lymphocyte counts were repeatedly measured in cattle living on three farms located in the “difficult‐to‐return zone” of the Fukushima nuclear accident, and compared with two control groups from unaffected areas. Blood cell counts differed significantly between the two control groups, although almost all the values fell within the normal range. The blood cell counts of the cattle in the “difficult‐to‐return zone” varied across sampling times even on the same farms, being sometimes higher or lower than either of the two control groups. However, neither a statistically significant decrease in blood cell counts nor an increase in the rate of cattle with extremely low blood cell counts was observed overall. The estimated cumulative exposure dose for the cattle on the most contaminated farm was within a range of 500–1000 mSv, exceeding the threshold for the lymphopenia. Because of the low dose rate on these farms, potential radiation damages would have been repaired and have not accumulated enough to cause deterministic effects.     

A comparison of an enzymatic and a gas-chromatographic method for measuring the acetate concentration in the blood plasma of cattle

The concentration of acetate in blood plasma was measured by both a gas-liquid chromatographic method and an enzymatic method using acetyl-CoA synthetase. When acetate was measured enzymatically without previous protein precipitation, the apparent concentration was lower than when the concentration measured by either a gas-chromatographic method or by the enzymatic method after protein precipitation with perchloric acid and neutralization.     

不同解离方法对莱克多巴胺猪尿阳性样品的提取效果研究

采用同位素内标法和高效液相-串联质谱法对猪尿莱克多巴胺阳性样品进行三种不同解离处理方法比较试验。结果表明,三种方法差异明显,β-葡萄糖醛苷酶/芳基硫酸酯酶在37℃酶解16 h效果最好,在60℃酶解2 h效果次之,0.1%高氯酸酸解提取效果最差。     

Estimating purine derivatives and nitrogen compound excretion using total urine collection or spot urine samples in grazing heifers

The study aimed to evaluate the excretion of purine derivatives (PDs) and nitrogen compounds (NCs) and their ratios with creatinine in supplemented Zebu heifers kept on pastures by comparing total urine collection and spot sampling. Five Nelore heifers (400 ± 15 kg) were used in a 5 × 5 Latin square design. The treatments were the amount of concentrate (220 g of crude protein/kg dry matter) offered (0, 3, 6, 9 and 12 g/kg BW). In each period, the total urine collection was performed continuously for 3 days (subsampled at intervals of 4 h, 00:00–04:00 h, 04:00–08:00 h, 08:00–12:00 h, 12:00–16:00 h, 16:00–20:00 h and 20:00–24:00 h). The spot urine samplings were performed (in each period) for 24 h (0, 4, 8, 12, 16 and 20 h). Creatinine, total urinary nitrogen (UN), urea nitrogen (UreaN), allantoin and uric acid were analysed. Creatinine excretion was 23.01 ± 0.19 mg/kg BW and was not affected by collection day, treatment or their interactions (p > 0.05). Treatments affected (p < 0.05) PD excretions, however did not affect the ratio PD:creatinine (p > 0.05). Treatments and collection time affected (p < 0.05) NC excretion, whereas the UN:creatinine and UreaN:creatinine ratios were not affected (p > 0.05). Creatinine excretion and the PD:creatinine ratios in the urine samples estimated by the total or spot sampling were not different (p > 0.05). However, sampling method affected (p < 0.05) the UN:creatinine and UreaN:creatinine (p < 0.05) ratios. Creatinine can adequately estimate urinary excretion in grazing heifers, and a single spot urine sample at any time of the day can be used to estimate PD excretion in grazing heifers. But two spot urine samples are needed for proper NC excretion estimations in grazing heifers’ urine.     

Identification of metoclopramide metabolites in the urine of cattle by gas chromatography-mass spectrometry and high-performance liquid chromatography-photodiode array detection

The urinary metabolites of metoclopramide (4-amino-5-chloro-N-[2-diethylaminoethyl]-2-methoxybenzamide) were identified in cows. The drug was administered intravenously, voided urine was collected, and individual urine extracts were analysed by gas chromatography-mass spectrometry and high-performance liquid chromatography-photodiode array detection. The parent compound and one major metabolite (4-amino-5-chloro-N-[2-(ethylamino)ethyl]-2-methoxybenzamide) were common to all individuals. In addition to the parent and major metabolite, a second, minor metabolite was identified in two cows as 4-amino-5-chloro-N-[2-(diethylamino)ethyl]-2-hydroxybenzamide. The identity of the minor metabolite was confirmed by comparison with a standard synthesized by a new method. Metabolite identification and characterization in food animal species allows the design of safety and environmental impact studies and relative metabolite ratios between dose treatment groups.Abbreviations ¹H-NMR proton nuclear magnetic resonance - R _T retention time - D₁ dopamine-1 - D₂ dopamine-2 - 5-HT₃ 5-hydroxytryptamine - GC-MS gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - UV-vis ultraviolet-visible - ID internal diameter - m/z mass/charge ratio     

反应堆污染部件放射性存留量估算方法

反应堆污染部件因结构复杂、条件苛刻等,其放射性存留量估算较为困难。本文基于测量和迭代模拟的思路建立了一套可行的估算方法,并对其估算原理、主要技术路线以及采用的计算机程序进行了描述。应用该估算方法,可对污染部件的放射性存留量进行有效估算。     

Effects of dietary crude protein and tannic acid on nitrogen excretion,urinary nitrogenous composition and urine nitrous oxide emissions in beef cattle

Two consecutive trials were carried out to study the effects of dietary crude protein (CP) and tannic acid (TA) on nitrogen (N) metabolism of beef cattle and consequently, the N₂O emissions from the urine of cattle. In Trial I, eight growing castrated cattle were used as the experimental animals. Two levels of dietary CP (110.6 and 135.7 g/kg dry matter [DM]) and two levels of TA (0 and 16.9 g/kg DM) were allocated in a replicated 2 × 2 crossover design. In Trial II, the N₂O emissions from the urine of cattle collected from Trial I were determined using the static incubation technique. An interaction between dietary CP and TA on the urinary N excretion (p < .05) was found but not on the N₂O‐N emission of cattle urine. Increasing dietary CP level from 110.6 g/kg DM to 135.7 g/kg DM increased the total N excretion (p < .001), the N retention (p < .05) and the ratio of urinary urea‐N/urinary N (p < .01), did not affect the N use efficiency (NUE; p > .05) and shifted the N excretion from faeces to urine. Increasing the dietary CP level increased the N₂O‐N emission of cattle urine. Dietary addition of TA decreased the urinary excretions of urea (p < .001) and shifted the N excretion from urine to faeces, did not affect the NUE of beef cattle (p > .10), and decreased the N₂O‐N emission of cattle urine. Pyrogallol and resorcinol of the TA metabolites were detected in urine with dietary addition of TA. Feeding beef cattle with relatively low CP level and adding TA in rations are effective approaches to mitigate the N₂O‐N emissions from cattle urine.     

Effects of urinary tract inflammation and sample blood contamination on urine albumin and total protein concentrations in canine urine samples

BACKGROUND: Urinary tract inflammation and hemorrhage are believed to be common causes of proteinuria in dogs based on results of studies that measured total urine protein concentration. A method to quantify urine albumin (UAlb) concentration in dogs recently has become available; however, the effect of inflammation on albuminuria is unknown. OBJECTIVES: The goals of this study were to determine the effects of urinary tract inflammation, as indicated by pyuria and sample blood contamination, on UAlb concentration and on urine protein:creatinine (UPC) ratio in dogs. METHODS: Urine samples were obtained from dogs with pyuria that were presented to a veterinary teaching hospital or were part of a laboratory colony. To mimic the effects of hematuria, canine whole blood was added to a microscopically normal canine urine sample that had baseline albumin and total protein concentrations below the limits of detection. UAlb concentration was measured using a canine albumin-specific competitive ELISA. UPC ratio was determined using routine methods. RESULTS: Of 70 samples with pyuria, 67% had negligible UAlb concentrations and 81% had normal UPC ratios. UAlb concentration but not UPC ratio was significantly higher (P < 0.05) in samples with concurrent hematuria or bacteriuria. When whole blood was added to normal urine, UAlb concentration did not exceed 1 mg/dL until the sample became visibly pink; the UPC did not exceed 0.4 at any dilution. CONCLUSIONS: Many dogs with pyuria do not have albuminuria or proteinuria; however, albuminuria may be more likely in dogs with pyuria and concurrent hematuria or bacteriuria. Hematuria may not cause an increase in UAlb concentration until it becomes macroscopic and even then may not increase the UPC ratio.     

Whole-genome QTL scan for ultrasound and carcass merit traits in beef cattle using Bayesian shrinkage method

Fine mapping of quantitative trait loci (QTL) for 16 ultrasound measurements and carcass merit traits that were collected from 418 hybrid steers was conducted using 1207 SNP markers covering the entire genome. These SNP markers were evaluated using a Bayesian shrinkage estimation method and the empirical critical significant thresholds (α = 0.05 and α = 0.01) were determined by permutation based on 3500 permuted datasets for each trait to control the genome-wide type I error rates. The analyses identified a total of 105 QTLs (p < 0.05) for seven ultrasound measure traits including ultrasound backfat, ultrasound marbling and ultrasound ribeye area and 113 QTLs for seven carcass merit traits of carcass weight, grade fat, average backfat, ribeye area, lean meat yield, marbling and yield grade. Proportion of phenotypic variance accounted for by a single QTL ranged from 0.06% for mean ultrasound backfat to 4.83% for carcass marbling (CMAR) score, while proportion of the phenotypic variance accounted for by all significant (p < 0.05) QTL identified for a single trait ranged from 4.54% for carcass weight to 23.87% for CMAR.     

Mixed venous and arterial blood in bovine coccygeal vessel samples for blood gas analysis

Abstract: Bovine coccygeal (median, caudal) vessel samples are not always venous in origin but may be arterial or a mixture of venous and arterial blood. Results of blood gas analysis of blood samples collected from 39 cows were consistent with typical venous, mixed venous-arterial, or typical arterial blood. This observation was made while establishing reference intervals for a new blood gas instrument. The pO₂ and the oxygen saturation of hemoglobin (SO₂) were most reflective of the amount of arterial blood contamination in the sample, but pCO₂ and pH also were altered. The pO₂ values ranged from 21 to 127 mm Hg, and SO₂ ranged from 35% to 100%. The mean pCO₂ in venous type samples was 46.3 mm Hg compared to 36.7 mm Hg in arterial type samples. The mean pH in venous type samples was 7.42 compared to 7.47 in arterial type samples.     

ECG and heart rate determination in fetal cattle using a digital signal processing method

A method of recording the fetal electrocardiogram (ECG) and heart rate (HR) is described. An abdominal lead and a thoracic lead were used to measure the fetal ECG and maternal ECG, respectively. The maternal component in the abdominal lead measurement was estimated by a digital adaptive filter using the thoracic lead measurement as a reference. By suppressing the estimated maternal components in the abdominal lead, the fetal ECG could be detected. The beat‐by‐beat fetal QRS complex peaks were determined by a digital matched filter from the fetal ECG enhanced in this way. The method was trialed in 10 pregnant Holstein cows at gestational periods ranging from 136 to 224 days. The results show that this method can extract the fetal ECG and determine the fetal HR from the raw noisy measurement data. The results suggest that the method would have applicability in monitoring fetal ECG as well as providing a non‐invasive, continuous HR profile during gestation, which will enable better understanding of the development of cattle fetuses before delivery.     

Quantification of pericaudal adipocyte diameter in dairy cattle during peripartum: a complementary method to study energetic status using conventional histology

The energetic status of high‐yielding Holstein‐Friesian dairy cattle was studied during peripartum under field conditions using body condition score (BCS), glycemia, seric ß–hydroxybutyrate and adipose tissue cellularity. This last method was tested as a complementary tool for energetic status assessment. Biopsies of pericaudal subcutaneous adipose tissue were obtained from 25 multiparous animals at 28 days before and 21 days after parturition. Samples were routinely processed for histological examination and stained with haematoxylin and eosin (H&E). The mean diameter of adipocytes (MDA) was measured with the aid of a digital image processor. During the same period, blood samples were collected weekly for metabolite determinations. The MDA at 28 days pre‐partum and 21 days post‐partum were 72.1 vs. 66.2 μm respectively (p = 0.055), and the corresponding BCS at these moments was 3.32 vs. 3.19 (p = 0.068). At ?28 days pre‐partum, the BCS was positively correlated with MDA (Pearson’s r = 0.521, p = 0.016) and with glycemia (Pearson’s r = 0.404, p = 0.056). Correlations between BCS and MDA, and between BCS and glycemia, with ß–hydroxybutyrate although not significant, suggest that routine histological preparations of biopsies from subcutaneous adipose tissue could be included as an easy and valuable tool for research purposes to evaluate metabolic adaptation of dairy cows to peripartum, as well as the incidence of metabolic disorders and productive performance.