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.     

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.     

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.     

A method for estimating radioactive cesium concentrations in cattle blood using urine samples

In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the ‘difficult‐to‐return zone’. Urine ¹³⁷Cs, specific gravity, electrical conductivity, pH , sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood ¹³⁷Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood ¹³⁷Cs] = [urinary ¹³⁷Cs]/([specific gravity] ? 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher ¹³⁷Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood ¹³⁷Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle.     

Estimation of concentration of radionuclides in skeletal muscle from blood,based on the data from abandoned animals in Fukushima

The damage caused by the earthquake on 11 March, 2011 resulted in a serious nuclear accident in Japan. Due to the damage to the Fukushima Daiichi Nuclear Power Plant (FNPP), large amounts of radioactive substances were released into the environment. In particular, one of the largest safety concerns is radioactive cesium (¹³⁴Cs and ¹³⁷Cs). Due to the FNPP nuclear accident, a 20 km area was restricted from human activity, and various types of domestic animals were left in the zone. We collected the organs and tissues from sacrificed animals to obtain scientific data to evaluate the internal deposition of radioactive compounds. At first, we found there is a strong correlation between blood ¹³⁷Cs and organ ¹³⁷Cs with data from 44 cattle, indicating that skeletal muscle is the target organ of deposition of radioactive cesium. Second, we analyzed the relationship between blood ¹³⁷Cs and muscle ¹³⁷Cs within relatively lower radioactive concentration, suggesting that estimation of concentration of ¹³⁷Cs is possible from blood concentration of ¹³⁷Cs. Finally, we developed computer software to estimate the muscle ¹³⁷Cs concentration from blood samples. Our study contributes to the food safety of livestock products.     

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.     

Effect of ammonium‐iron‐hexa‐cyanoferrate and of the covariates age,gender, weight,season and calendar time on radiocaesium contamination of wild boars living in the wild in Bavaria

Feed with Ammonium‐iron‐hexa‐cyanoferrate (AFCF; 1250 mg AFCF/kg) was fed between March 2009 and March 2011 to wild boars in a territory of 4.5 km² (experimental group, EXP). One hundred and forty similar territories in the same county (500 km², spruce forest, agriculture) served as control (CON). Data for comparison from all territories were available from March 2005 to March 2011. Wild boars could move between, into and from the territories. Lean skeletal muscle meat (500 g) of all wild boars that were killed by humans (hunting and traffic accidents) was investigated for gamma‐radiation from ¹³⁷Cs with a becquerel monitor with a sodium iodide scintillator crystal (range of detection 20–9999 Bq/kg). The wild boars were weighed, and gender and age were determined. For the analyses of effects, multivariable regression models were fitted with the ¹³⁷Cs concentration as response variable. There was a significant difference between the ¹³⁷Cs contamination of wild boars from CON (563 ± 932 Bq/kg meat, n = 1253) and EXP (236 ± 276 Bq/kg meat; n = 45). ¹³⁷Cs contamination decreased with increasing body weight by ?5 Bq/kg meat/kg body weight increase (p < 0.05). Females had higher Bq measurements than males (by +80 Bq/kg meat, p < 0.05). Piglets were lower than adults, but turn‐coats higher. From November to May, contamination was higher (by +500 to +600 Bq/kg meat, p < 0.05) than during the rest of the year. In 2010, contamination was higher (by +200 to + 300 Bq/kg meat, p < 0.05) in comparison with the other years under observation. When all covariates were controlled for, the effect of AFCF was highly significant. Interaction analyses showed that the intervention decreased ¹³⁷Cs contamination by ?500 Bq/kg meat during November to May and by ?200 Bq/kg meat during the rest of the year. In summary, AFCF feeding reduces ¹³⁷Cs contamination of wild boars living in the wild significantly, particularly during the season from November to May.     

A Rapid Method for the Determination of Radioactive Caesium in Live Animals and Carcasses,and its Practical Application in Norway after the Chernobyl Nuclear Reactor Accident

A technique was developed for the measurement of levels of caesium radionuclides (¹³⁷Cs+¹³⁴Cs) in live reindeer, cattle, and sheep and in carcasses from these species. The instrument used was a sodium iodide scintillation detector coupled to a portable multi-channel analyser.Based on a combination of background measurements and measurements of impulses from animals with the detector in different anatomical positions we recommmend the following procedures: Lamb: The detector placed on os sacrum (standing animal). Reindeer: The detector placed between the hind legs (animal lying on its side). Cattle: The detector placed on the back of the standing animal, midway between os sacrum and trochanter major.Average geometrical factors for live animals were estimated. It was a linear correlation between measured activity levels in meat samples and counted impulses per sec in live animals. Geometrical factors were estimated at 95% confidence level with uncertainty between 6–14%. The detection limits varied between 50–200 Bq (becquerel)/kg in areas with ground depositions between 5–200 kBq/m². Since the winter 1986/87 the technique has been the standard procedure for monitoring slaughter animals and carcasses for radiocaesium activity concentrations.     

The Influence of a »Flushing Dose« of Selenium on the Excretion of Selenium in Sheep

The influence of injected selenium on the excretion of radioactive selenium stored in the body was studied in 4 male sheep. At the beginning of the experiment, radioactive selenium as Se⁷⁵-sodium selenite and Se⁷⁵-L-selenomethionine was injected subcutaneously, each compound in 2 rams. The injection of Se⁷⁵-sodium selenite was repeated in one ram after 37 days. Non-radioactive selenium as sodium selenite or DL-selenomethionine was injected between 5 and 47 days after the injection of the Se⁷⁵-dose. Each injection of non-radioactive sodium selenite increased the urinary Se⁷⁵-excretion on an average 10 times. A close correlation was demonstrated between the concentration of Se⁷⁵ and total selenium in urine.After the subcutaneous injection of Se⁷⁵-L-selenomethionine the Se⁷⁵-excretion in urine was increased both with a “flushing dose” of sodium selenite and with DL-selenomethionine. The concentration of Se⁷⁵ and that of total selenium in urine of these animals were closely correlated.Se⁷⁵-excretion in feces was not markedly influenced by the administration of non-radioactive selenium.     

Tissue distribution of albumin‐glucose advanced glycation end products administrated to chickens

Glycation (Maillard reaction) starts from non‐enzymatic amino‐carbonyl reaction binding carbonyl group of reducing sugars to amino group of amino acids. The amino‐carbonyl reaction leads to the formation of the stable Amadori product, which ultimately forms advanced glycation end products (AGE). Acceleration of glycation during hyperglycemia increases the production and accumulation of AGE, which is implicated in the gradual development of diabetic complications in diabetes mellitus. Avian species offers the great advantage of providing animal models for diabetology because high blood glucose concentration should accelerate to generate high concentration of AGE. In the present study, radioactive AGE was prepared from chicken serum albumin and ¹⁴C‐glucose. Radioactive AGE was administrated to examine the tissue distribution of AGE in chickens. Administration of radioactive AGE into the circulation of chickens revealed that AGE accumulated into specific tissues, liver, spleen and kidney, which have the function for clearing exogenous substances and endogenous metabolic waste products.     

Preliminary Studies on the Concentration of Na+,K+-ATPase in Skeletal Muscle of Draught Cattle in Mozambique: Effect of Sex,Age and Training

The effect of training on the potential for work in draught cattle was assessed by measuring the Na⁺,K⁺-ATPase in the muscle cell membrane and the elevation in the concentration of K⁺ in plasma during exercise. Biopsies of the semitendinosus muscle and venous blood samples were taken from the cattle used for draught work in Mozambique. No differences were found in the plasma ion or Na⁺,K⁺-ATPase concentrations in samples taken from Nguni, Africander and Angoni breeds. There were no significant differences in plasma ions (Na⁺, K⁺ and Cl^–) or muscle Na⁺,K⁺-ATPase concentrations between the Angoni males and females, although the males showed an increase in Na⁺,K⁺-ATPase with age, while the females showed a decrease. The increase in males might be attributed to their higher level of activity in the herds than that of females. After a training period of 15 days, a significant increase in Na⁺,K⁺-ATPase concentration in semitendinosus muscle was found in Angoni cattle. In females, this was significant after 8 days of training (about 30%); in males after 15 days of training (about 16%). On day 15, there was a reduction in the elevation of plasma K⁺ during the 2 h of draught work, indicating an increased capacity of the Na⁺,K⁺ pumps to maintain the extracellular K⁺ concentration in working muscles and a possible delay in the moment of fatigue.     

Occurrences of thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> in cattle slaughtered at Morogoro municipal abattoir,Tanzania

An investigation was conducted in Morogoro municipality to assess the likelihood of slaughter cattle posing public health risk of contaminating carcasses with thermophilic Campylobacter. Butchers and meat shopkeepers were interviewed on source of slaughter cattle, method of animal and carcass transportation, carcass dressing, meat storage facilities, access to clean water and availability of food hygiene practices. Faecal samples were collected from 107 slaughter cattle and after slaughter; four different parts of dressed carcasses (i.e. from ham, neck, pelvis and thigh muscles) were also sampled. In addition 107 cattle meat samples for Campylobacter culture were collected in different meat shops. The samples were subjected to standard bacteriological examination using Skirrows protocol. It was found that cattle slaughter, dressing and meat handling in meatshops was done under unhygienic condition. Thermophilic Campylobacter prevalence in slaughter cattle was 5.6% while contamination rate of dressed carcasses and cattle meat at shops was 9.3% and 1.9%, respectively. The majority of thermophilic Campylobacter isolated were C. jejuni (88.9%) while C. coli was isolated at 11.1%. Findings of this study suggest possibility of humans acquiring zoonotic Campylobacter infections from cattle meat particularly when meat preparation and processing is not done properly. More work is required to establish the magnitude of zoonotic enteric Campylobacteriosis in humans and epidemiological role of cattle and other animals in the study area.     

Morphological reproductive characteristics of testes and fertilization capacity of cryopreserved sperm after the Fukushima accident in raccoon (Procyon lotor)

Since the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident, we have established an archive system of livestock and wild animals from the surrounding ex-evacuation zone. Wildlife within the alert zone have been exposed to low-dose-rate (LDR) radiation for a long continuous time. In this study, we analysed the morphological characteristics of the testes and in vitro fertilization (IVF) capacity of cryopreserved sperm of racoons from the ex-evacuation zone of the FDNPP accident. The radioactivity of caesium-137 (¹³⁷Cs) was measured by gamma-ray spectrometry, and the measured radioactivity concentration was 300–6,630 Bq/kg in the Fukushima raccoons. Notably, normal spermatogenesis was observed in the seminiferous tubules of the testes, with the germinal epithelium composed of a spermatogenic cell lineage with no evident ultrastructural alterations; freeze-thawing sperm penetration ability was confirmed using the interspecific zona pellucida-free mouse oocytes IVF assays. This study revealed that the chronic and LDR radiation exposure associated with the FDNPP accident had no adverse effect on the reproductive characteristics and functions of male raccoons.     

An epidemiological survey on bovine and ovine babesiosis in Kurdistan Province, western Iran

The aim of this study was to determine the prevalence of the Babesia infection in domestic animals in Kurdistan Province of Iran for the first time. In this survey, 9,111 domestic livestock, including cattle and sheep, were randomly sampled and examined from 500 flocks in Kurdistan Province from July 2007 to September 2009. Thin peripheral blood smears were taken and then stained by Giemsa staining method. From a total of 9,111 collected samples, 2,642 were sheep and 6,469 were cattle. Babesia spp. is detected in 1,359 (51.4%) out of sheep samples and 136 (2.1%) out of cattle samples by direct examination of blood smear. Altogether, the prevalence rate of Babesia infection was 16.4% (n = 1,495) in both animal groups. Babesia ovis and Babesia bigemina were the most prevalent species found in sheep and cattle, respectively. The relatively high prevalence of Babesia infection in livestock indicates the epizootic stability status of babesiosis in the western part of Iran.     

Decomposition of cattle dung on grazed signalgrass (Brachiaria decumbens Stapf) pastures in monoculture or intercropped with tree legumes

Livestock excreta is one of the major nutrient sources in natural grasslands. Understanding how livestock diet and season affects the decomposition dynamics is critical to nutrient cycling models. We hypothesised that livestock diet and season of the year affect dung decomposition. This study evaluated the decomposition and release of N, P, K, Ca, Mg, and Na from faeces of cattle collected in dry and wet seasons. Treatments were signalgrass (Brachiaria decumbens Stapf) in monoculture or mixed with sabiá (Mimosa caesalpiniifolia Benth.) or gliricidia (Gliricidia sepium (Jacq.) Kunth ex Walp.). Excreta samples were incubated in nylon bags for 0, 2, 4, 8, 16, 32, 64, 128 and 256 d. The single negative exponential mathematical model was adequate (P < 0.0001) to explain the decomposition. The relative rate of dung decomposition (k) was greater for samples that originated from cattle grazed on signalgrass (k = 0.00284 g g^?1 d^?1), followed by sabiá (k = 0.00233 g g^?1 d^?1), and gliricidia (k = 0.00200 g g^?1 d^?1) pastures. The rate of nutrient release showed a time effect for all variables and interaction between time and season for Ca (P = 0.0042) and Mg (P = 0.0013). Faeces collected from cattle grazing/browsing intercropped pastures tended to have lower decomposition rates.     

Developing high throughput quantitative PCR assays for diagnosing Ikeda and other Theileria orientalis types common to New Zealand in bovine blood samples

AIMS: To develop rapid, quantitative PCR (qPCR) assays using high resolution melt (HRM) analysis and type-specific TaqMan assays for identifying the prevalent types of Theileria orientalis found in New Zealand cattle; and to evaluate their analytical and diagnostic characteristics compared with other assays for T. orientalis.

METHODS: Nucleotide sequences aligned with T. orientalis Buffeli, Chitose and Ikeda types, obtained from DNA extracted from blood samples from infected cattle, were used to design HRM and type-specific probe-based qPCR assays. The three type-specific assays were also incorporated into a single-tube multiplex qPCR assay. These assays were validated using DNA extracted from blood samples from cattle in herds with or without clinical signs of T. orientalis infection, other veterinary laboratory samples, as well as plasmids containing T. orientalis type-specific sequences. Diagnostic specificity (DSp) and sensitivity (DSe) estimates for the qPCR assays were compared to blood smear piroplasm results, and other PCR assays for T. orientalis. Copy number estimates of Ikeda DNA in blood were determined from cattle exhibiting anaemia using the Ikeda-specific qPCR assay.

RESULTS: The T. orientalis type-specific and the HRM qPCR assays displayed 100% analytical specificity. The Ikeda-specific qPCR assay exhibited linearity (R²?=?0.997) with an efficiency of 94.3%. Intra-assay CV were ≤0.08 and inter-assay CV were ≤0.095. For blood samples from cows with signs of infection with T. orientalis, the DSp and DSe of the multiplex probe qPCR assay were 93 and 96%, respectively compared with blood smears, and 97 and 100%, respectively compared with conventional PCR assays. For the Ikeda-specific qPCR assay, the number of positive samples (n=66) was slightly higher than a conventional PCR assay (n=64). The concentration of Ikeda genomes in blood samples from 41 dairy cows with signs of infection with T. orientalis ranged between 5.6?×?10⁴ and 3.3?×?10⁶ genomes per µL of blood.

CONCLUSIONS: All qPCR assays had improved specificity and sensitivity over existing conventional PCR assays for diagnosis of T. orientalis Ikeda. The burden of Ikeda DNA in blood was demonstrated using an Ikeda-specific qPCR assay with titrated synthetic gene target.

CLINICAL RELEVANCE: Adoption of high-throughput DNA extraction and qPCR reduced T. orientalis and Ikeda diagnosis times. The Ikeda-specific qPCR assay provides a specific diagnosis for Ikeda in animals with signs of infection with T. orientalis and can be used to monitor the parasite load of Ikeda in blood.     

Development of an ELISA system for Detection of Anti‐Anaplasma marginale Antibodies in cattle in Brazil

An ELISA test was developed for detecting antibodies against Anaplasma marginale in bovine sera. Four antigenic preparations were produced from infected red blood cells. Some aliquots of this preparation were stored at ‐70°C with 30% DMSO in phosphate‐buffered saline (PBS) and others were lysed with 0.9% NH₄Cl and stored at ‐ 20°C. Typical anaplasmal structures were seen by electron microscopy in the antigenic preparations containing the erythrocytes that had been stored with DMSO. The performance of the ELISA test was evaluated by testing 298 positive serum samples collected from immunized cattle, 39 negative serum samples collected from cattle imported from areas free of A. marginale and 50 samples collected from cattle naturally infected in the field. The test gave a specificity of 94.87% and a sensitivity of 100%.     

Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial‐Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms

The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial‐resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross‐sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial‐resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter‐specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between livestock and wildlife samples, and the lack of similarity in molecular subtypes and AMR patterns, we concluded that the sharing of Campylobacter species between livestock and mammalian wildlife was uncommon.