Seasonal differences in the intensity of acute phase response in dogs infected with Babesia canis.

The highest number of acute Babesia canis cases in dogs is recorded over the February-May (Feb-May) period, which also represents the optimal climate conditions for tick activity in Belgrade, Serbia. A possibility that the acute phase response is more intense in dogs developing the disease in the Feb-May period compared with the response in other time periods of the year was tested. A total of 63 client-owned dogs with acute B. canis infection were enrolled and the routine hematology and biochemistry parameters-serum amyloid A (SAA), IgG against B. canis, level of parasitemia, ceruloplasmin (CER), paraoxonase-1 (PON-1), and fibrinogen-were measured. Acute phase indexes (API) were calculated as (SAA×CER)/(Iron×PON-1) and (SAA×CER)/(Albumin×Iron). Statistics included Kruskal-Wallis test and logistic regression analysis. The results showed that in the Feb-May period, the following parameters were lower: creatinine, albumin, iron, and level of parasitemia. Furthermore, increased API values were more probable in the Feb-May than in the other periods. Together, higher acute phase response intensity and presumptive hemodilution in the Feb-May period indicate a more acute course of B. canis infection than in other time periods of the year.

Systemic inflammatory response syndrome in dogs naturally infected with Babesia canis: Association with the parasite load and host factors.

The common signs of canine babesiosis caused by an infection with Babesia canis are fever, anorexia, lethargy, pulse alterations, anemia, and occasionally mild icterus. Dogs with these clinical signs can be divided into two groups: those with acute-phase reaction and those with systemic inflammatory response syndrome (SIRS). Factors associated with the occurrence of SIRS in canine babesiosis have not been thoroughly researched. This article outlines a cross-sectional study of 54 client-owned dogs with an acute B. canis infection, and evaluates the differences in age, gender, laboratory findings, parasite load, and seroreactivity against B. canis between the SIRS and the SIRS-free dogs. We have analyzed a complete blood count, serum biochemistry, serum amyloid A, ceruloplasmin, paraoxonase-1, serology, and PCR testing using standard methodologies. The frequency of SIRS among the investigated dogs reached 0.59. Male dogs and those seronegative against B. canis, were more frequent in the SIRS group, whilst age and parasite load could not be associated with the presence of SIRS. Dogs with SIRS had a lower count of total leukocytes, neutrophils, lymphocytes, and monocytes, and a lower concentration of iron and bilirubin compared with SIRS-free dogs. No significant differences in the concentration of acute-phase proteins have been noticed to exist between the groups of dogs. Further, the seronegative dogs had a lower count of lymphocytes and monocytes and a higher parasite load than the seroreactive dogs. Multivariate logistic regression analysis has identified leukopenia (<6 × 109/L) and monocytopenia (<0.2 × 109/L) as independent associates of SIRS in the investigated dogs, thus implying that these routine tests could be used as reliable markers for SIRS.

Evidence of acute phase reaction in asymptomatic dogs naturally infected with Babesia canis.

Asymptomatic outdoor dogs can be carriers of Babesia canis, but data describing the development of an acute phase response (APR) are not available. We hypothesised that these dogs have a moderate APR that could be detected by hematological and biochemical changes. Two groups of Babesia-exposed dogs were represented by nine B. canis PCR-positive and twenty B. canis PCR-negative, seroreactive dogs. The control group consisted of ten Babesia-naïve dogs. Serum amyloid A (SAA), paraoxonase-1 (PON-1), complete blood count, and biochemistry parameters were analysed by standard methodologies. Protein and lipoprotein fractions were separated using agarose gel electrophoresis (GE), and the dominant diameters of lipoproteins were assessed on gradient GE. Results were evaluated using non-parametric tests and the Receiver Operating Characteristic curve. SAA (median 39.0 µg/mL, range 2.2-48.8 µg/mL), total protein (median 74.7 g/L, range 57.1-98.3 g/L) and the dominant diameter of α-lipoproteins (median 13.31 nm, range 12.09-14.17 nm) in B. canis PCR-positive dogs were higher relative to dogs in the control group or dogs that were PCR-negative but seroreactive (p < 0.001 for both groups). Mild to moderate anemia (4/29), thrombocytopenia (7/29), and leukocyte counts that were close to the upper limit of the reference range were encountered in both Babesia-exposed groups. When compared to controls, Babesia-exposed dogs displayed decreased a PON-1 activity and protein GE pattern consistent with low-grade chronic inflammation (p < 0.001 for both groups). Dogs with detectable amounts of B. canis DNA in blood contain increased levels of SAA and total protein along with α-lipoproteins that display an increased diameter relative to those dogs with positive Babesia serology but undetectable levels of B. canis DNA in blood.

Presence of natural radionuclides and toxic elements in monocalcium phosphate, complete feed and pig manure.

Naturally occurring radionuclides and toxic elements are ubiquitous in the environment and can pose a great risk to animals' health. Control of these toxic substances in animal diet is essential for animal's welfare, human nutrition risk management as well as environmental protection. Our study addresses these issues by determining a content of natural radionuclides (40K, 238U and 226Ra), and toxic elements (Cd, Cr, As) in total of 108 samples: 36 monocalcium phosphate, 36 complete feed and 36 pig manure samples that were collected from 6 different commercial farms in the Republic of Serbia. In monocalcium phosphate the content of 40K, 238U and 226Ra ranged from 7.6-23.4 Bq kg-1, 13.2-1778 Bq kg-1 and 3.6-13.5 Bq kg-1, respectively. In complete feed samples, 40K ranged from 170 to 306 Bq kg-1, 238U from minimum detectable activity (MDA) to 9.1 Bq kg-1, and 226Ra from MDA-5.5 Bq kg-1, while in pig manure 40K ranged from 146 to 531 Bq kg-1, 238U from MDA-30.1 Bq kg-1, and 226Ra from MDA-10 Bq kg-1. The concentrations of Cd, Cr and As in monocalcium phosphate were 0.26-14.9 mg kg-1, 10.3-497 mg kg-1, and 4.2-5.5 mg kg-1, respectively. In complete feed, the content of Cd, Cr and As ranged from minimum detectable concentration (MDC) - 0.78 mg kg-1, 0.30-7.41 mg kg-1, and 0.09-0.31 mg kg-1, respectively. In pig manure Cd, Cr and As ranged from 0.8-2.3 mg kg-1, 2.3-22.2 mg kg-1, and 0.51-5.45 mg kg-1, respectively. The obtained results showed that Cd concentration in monocalcium phosphate exceeded the maximum permissible legislative concentrations for phosphate additives. Monocalcium phosphate is the main source of 238U and Cr in complete feed, while Cd derives from other sources. For that reason excessive use of monocalcium phosphate can lead to high levels of these pollutants in complete feed and consequently in manure.

Characterization of personal solar ultraviolet radiation exposure using detrended fluctuation analysis.

Studies of personal solar ultraviolet radiation (pUVR) exposure are important to identify populations at-risk of excess and insufficient exposure given the negative and positive health impacts, respectively, of time spent in the sun. Electronic UVR dosimeters measure personal solar UVR exposure at high frequency intervals generating large datasets. Sophisticated methods are needed to analyze these data. Previously, wavelet transform (WT) analysis was applied to high-frequency personal recordings collected by electronic UVR dosimeters. Those findings showed scaling behavior in the datasets that changed from uncorrelated to long-range correlated with increasing duration of time spent in the sun. We hypothesized that the WT slope would be influenced by the duration of time that a person spends in continuum outside. In this study, we address this hypothesis by using an experimental study approach. We aimed to corroborate this hypothesis and to characterize the extent and nature of influence time a person spends outside has on the shape of statistical functions that we used to analyze individual UVR exposure patterns. Detrended fluctuation analysis (DFA) was applied to personal sun exposure data. We analyzed sun exposure recordings from skiers (on snow) and hikers in Europe, golfers in New Zealand and outdoor workers in South Africa. Results confirmed validity of the DFA superposition rule for assessment of pUVR data and showed that pUVR scaling is determined by personal patterns of exposure on lower scales. We also showed that this dominance ends at the range of time scales comparable to the maximal duration of continuous exposure to solar UVR during the day; in this way the superposition rule can be used to quantify behavioral patterns, particularly accurate if it is determined on WT curves. These findings confirm a novel way in which large datasets of personal UVR data may be analyzed to inform messaging regarding safe sun exposure for human health.

A short-term and long-term relationship between occurrence of acute canine babesiosis and meteorological parameters in Belgrade, Serbia.

Belgrade, the capital of the Republic of Serbia, is an endemic location for canine babesiosis caused by Babesia canis. This tick-borne disease occurs seasonally in regions with moderate continental climate. However, recent clinical data show that: 1) some cases of canine babesiosis have been recorded during the winter, and 2) canine babesiosis is spreading to the northern parts of Europe with a colder climate, which is a region previously free of this disease. Our study investigates the occurrence of canine babesiosis in different seasons over 2013-2016, and explores a short-term link between meteorological parameters and number of diagnosed cases of canine babesiosis in Belgrade. It also looks into possible long-term correlations that span one year before the onset of the disease. Based on 872 recorded cases over four years, our results show a bimodal seasonal distribution of canine babesiosis, with a pronounced peak in the spring, and a less conspicuous one in the autumn. Throughout the year, even over the coldest and warmest periods, there is a broad range of temperatures and relative humidities when the disease is recorded. Over one year prior to the spring and autumn onset of the disease, we found a noticeable impact of temperature and relative humidity, and to a lesser extent, of atmospheric pressure and cloud cover, on the number of diagnosed cases. These findings imply short-term and long-term relationships between occurrence of acute canine babesiosis and certain meteorological parameters, and they open further questions that need to be investigated in order to understand the epidemiology of this disease.

Analysis of trends, periodicities, and correlations in the beryllium-7 time series in Northern Europe.

The activity concentrations of beryllium-7, a natural radiotracer that is considered as a tracer of the stratospheric-tropospheric exchange, shows a distinct behaviour in Northern Europe compared to the central and southern parts of the continent. In this study, we use the measurements collected at four sampling stations in Scandinavia (Ivalo, Umea, Kista, Risoe) between 2001 and 2010 and investigate their trends, periodicities and residuals with the aim to further understand the common features in the beryllium-7 data records in northern sampling sites. The beryllium-7 activity concentrations exhibit statistically significant positive trends that range from an average value of 1.50%/year to an average value of 2.96%/year. We detect a one-year periodicity in all the sites, and in the southernmost site, Risoe in Denmark, additional higher-frequency harmonics. In the residual time series, we find outliers that represent occurrences of extremely high beryllium-7 activity concentration. Cross-correlations of the beryllium-7 residuals across the four sites decrease with increasing distance; similarly, as indicated by local Hurst exponents the records exhibit long-range correlations that weaken towards the end of the investigated period. To investigate the causes of the detected trends, we also calculate correlations between beryllium-7 and factors related to its production, transport and removal from the atmosphere: in particular, cross-correlations of the beryllium-7 residuals with residuals in sunspot number, local temperature, atmospheric pressure and precipitation, as well as Arctic Oscillation index and North Atlantic Oscillation index. Most of the obtained correlations, however, are not statistically significant, highlighting the need to analyse a longer time period in order to evaluate the impact of different factors on the airborne beryllium-7 activity concentration.

Novel approach to analysing large data sets of personal sun exposure measurements.

Personal sun exposure measurements provide important information to guide the development of sun awareness and disease prevention campaigns. We assess the scaling properties of personal ultraviolet radiation (pUVR) sun exposure measurements using the wavelet transform (WT) spectral analysis to process long-range, high-frequency personal recordings collected by electronic UVR dosimeters designed to measure erythemal UVR exposure. We analysed the sun exposure recordings of school children, farmers, marathon runners and outdoor workers in South Africa, and construction workers and work site supervisors in New Zealand. We found scaling behaviour in all the analysed pUVR data sets. We found that the observed scaling changes from uncorrelated to long-range correlated with increasing duration of sun exposure. Peaks in the WT spectra that we found suggest the existence of characteristic times in sun exposure behaviour that were to some extent universal across our data set. Our study also showed that WT measures enable group classification, as well as distinction between individual UVR exposures, otherwise unattainable by conventional statistical methods.

Natural and anthropogenic radioactivity in the environment of Kopaonik mountain, Serbia.

To evaluate the state of the environment in Kopaonik, a mountain in Serbia, the activity concentrations of (4) K, (226)Ra, (232)Th and (137)Cs in five different types of environmental samples are determined by gamma ray spectrometry, and radiological hazard due to terrestrial radionuclides is calculated. The mean activity concentrations of natural radionuclides in the soil are higher than the global average. However, with an exception of two sampling locations, the external radiation hazard index is below one, implying an insignificant radiation hazard. Apart from (40)K, content of the natural radionuclides is predominantly below minimum detectable activities in grass and cow milk, but not in mosses. Although (137)Cs is present in the soil, grass, mosses and herbal plants, its specific activity in cow milk is below minimum detectable activity. Amongst the investigated herbal plants, Vaccinium myrtillus L. shows accumulating properties, as a high content of (137)Cs is detected therein. Therefore, moderation is advised in consuming Vaccinium myrtillus L. tea.

Leaves of higher plants as biomonitors of radionuclides (137Cs, 40K, 210Pb and 7Be) in urban air.

Leaves of linden (Tilia tomentosa L. and Tilia cordata Mill.) and horse chestnut (Aesculus hippocastanum L.) were analysed as biomonitors of radionuclides in urban air. Samples of soils, leaves and aerosols were collected in Belgrade, Serbia. Activities of (137)Cs, (40)K, (210)Pb and (7)Be in the samples were measured on an HPGe detector by standard gamma spectrometry. "Soil-to-leaves" transfer factors were calculated. Student's t test and linear Pearson correlation coefficients were used for statistical analysis. Differences in local conditions at the sampling sites were not significant, and the mechanisms of the radionuclides' accumulation in both plant species are similar. Ceasium-137 was detected in some of the leaf samples only. Transfer factors for (137)Cs and (40)K were (0.03-0.08) and 1.3, respectively. The concentrations of (210)Pb and (7)Be in leaves were higher in autumn than in spring, and there were some similarities in their seasonal patterns in leaves and in air. Weak to medium correlation was obtained for the (210)Pb and (7)Be activities in leaves and aerosols. Large positive correlation was obtained for the (210)Pb activities in linden leaves and the mean activity in aerosols for the preceding months. Different primary modes of radionuclides accumulation in leaves were observed. Since large positive correlation was obtained for the (210)Pb activity in linden leaves and the mean in aerosols for the preceding months, mature linden leaves could be used as biomonitors of recent (210)Pb activity in air.

Radioactivity monitoring in ground level air in Belgrade urban area.

Concentrations of beryllium-7 ((7)Be), lead-210 ((210)Pb) and caesium-137 ((137)Cs) were measured at two sites in the city of Belgrade (Serbia). One monitoring site was located in the central city area with heavy traffic and the other within the Institute of Nuclear Sciences Vinca, in the suburbs. Presented data cover the period 2004-9. Activity of the radionuclides was determined on an HPGe detector (Canberra, relative efficiency 23 %) by standard gamma spectrometry. Activities of (7)Be and (210)Pb exhibit a similar seasonal pattern. The mean monthly concentrations of (7)Be did not exceed 7 mBq m(-3). The maxima are correlated with the seasonal increase in temperature, whereas the minima are linearly correlated with the amount of precipitation. The activity of (210)Pb was below 0.9 mBq m(-3). The activities of both radionuclides were very low in winters and were largely affected by precipitation and snow coverage. Concentrations of (137)Cs were mainly below the limit of detection.

Concentration of trace elements in blood and feed of homebred animals in Southern Serbia.

BACKGROUND, AIM AND SCOPE: The paper presents concentrations of trace elements in blood of homebred animals (cows and sheep) from Southern Serbia (Bujanovac) and the contents of natural and anthropogenic radionuclides and some heavy metals in feed. The region of Southern Serbia was exposed to contamination by depleted uranium ammunition during NATO attacks in 1999 and therefore, is of great concern to environmental pollution and human and animal health. MATERIALS AND METHODS: Conventional instrumental and epithermal neutron activation analyses are used to measure trace elements in cow and sheep blood samples collected randomly at six locations in the region of Bujanovac (village of Borovac) in the spring of 2005. Samples of feed (grass and crops: corn, wheat and oats), collected on the same locations (households), are analysed for the contents of radionuclides on an HPGe detector (Ortec, relative efficiency 23%) by standard gamma spectrometry. The content of Hg, Pb and Cd in feed is determined by standard atomic absorption spectrometry on the VarianSpectra220/ThermoSolar GFS97 spectrometer. RESULTS: Concentrations of 29 elements (Na, Al (P), Cl, K, Sc, Cr, Mn, Ni, Fe, Co, Zn, Se, As, Br, Sr, Rb, Sb, In, I, Ba, Cs, La, Nd, Eu, Sm, Tb, Hf, Ta and Th) are determined in blood of the examined animals. In feeds, natural (40)K is found in all of the samples, cosmogenic (7)Be and fission product (137)Cs are detected only in the grass samples, while heavy metals Hg, Cd and Pb are found in the range of 0.01-0.02, 0.84-1.15 and 0.74-7.34 mg/kg, respectively. Calculated soil-to-blood transfer factors are in a wide range of 8 x 10(-6) to 64, as a result of varying significance of the elements in animal metabolism and feeding habits. DISCUSSION: The results of trace elements' concentrations in animal blood are in good agreement with available data for K, Ni, Zn, Se and Rb. Higher Br concentrations in animal blood are most probably caused by large biomass burning events during blood sampling. Very low concentration of Fe in cows and sheep confirms the results of previous biochemical studies on animal anaemia in the region. High concentration of As correlates with geochemical peculiarities of the Balkans and is also likely influenced by the use of pesticides in the agricultural production. For some of the elements (La, Nd, Eu, Sm, Tb, Sb, Hf, Ta, Th, In, Ba, Sr, Sc and Cs), there are few or no literature data. Therefore, some of the presented data are significant not only for the country and the region, but on a wider scale. Activities of natural radionuclides in feeds are within the average values reported for the region, while the activities of (210)Pb and (235/238)U are below the limit of detection. This is in accordance with previous investigations showing no widespread contamination by depleted uranium in the area. Contents of Hg and Pb in feeds are below the nationally permissible levels, unlike the content of Cd which exceeds it, probably caused by the use of phosphate fertilisers and fossil fuel combustion in the area. CONCLUSIONS: In general, the concentrations of trace elements in blood of homebred cows and sheep are in good agreement with reference materials, available literature data and the results of previous studies in the area. The exceptions are Fe, As and Br. The contents of natural and anthropogenic radionuclides in feeds are within the expected levels, and there are no signs of contamination by depleted uranium or other fission products. Apart from Cd, there are no signs of pollution by heavy metals in feeds. The highly sensitive method of instrumental neutron activation analysis provides data on the concentration of some elements in animal blood not previously reported for the region and elsewhere. RECOMMENDATIONS AND PERSPECTIVES: The presented study is a part of the long term ongoing project on the health risk assessment on animals and humans in the region. The collected data is intended to provide a base for the animal and human risk assessment as well as an estimate of the general pollution status of the environment in the region. Since some of the investigated elements are classified as important trace elements for livestock, the results could also be used to balance and improve the animal diet and thus, improve the growth and reproduction rate.

Radionuclides and heavy metals in Borovac, Southern Serbia.

BACKGROUND, AIM, AND SCOPE: The paper presents the complex approach to the assessment of the state of the environment in Southern Serbia, surroundings of Bujanovac, the region which is of great concern as being exposed to contamination by depleted uranium (DU) ammunition during the North Atlantic Treaty Organization (NATO) attacks in 1999. It includes studies on concentrations of radionuclides and heavy metals in different environmental samples 5 years after the military actions. MATERIALS AND METHODS: In October 2004, samples of soil, grass, lichen, moss, honey, and water were collected at two sites, in the immediate vicinity of the targeted area and 5 km away from it. Radionuclide ((7)Be, (40)K, (137)Cs, (210)Pb, (226)Ra, (232)Th, (235)U, (238)U) activities in solid samples were determined by standard gamma spectrometry and total alpha and beta activity in water was determined by proportional alpha-beta counting. Concentrations of 35 elements were determined in the samples of soil, moss, grass, and lichen by instrumental neutron activation analysis (INAA). RESULTS: The results are discussed in the context of a possible contamination by DU that reached the environment during the attacks as well as in the context of an environmental pollution by radionuclides and heavy metals in Southern Serbia. The results are compared to the state of environment in the region and other parts of the country both prior to and following the attacks. DISCUSSION: This is the first comprehensive study of the contents of radionuclides and heavy metals in Southern Serbia and consequently highly important for the assessment of the state of environment in this part of the country concerning possible effects of DU ammunition on the environment, as well as anthropogenic source of pollution by radionuclides and heavy metals and other elements. Also, the highly sensitive method of INAA was used for the first time to analyze the environmental samples from this area. CONCLUSIONS: The results of the study of radionuclides in the samples of soils, leaves, grass, moss, lichen, honey, and water in Southern Serbia (Bujanovac) gave no evidence of the DU contamination of the environment 5 years after the military actions in 1999. Activities of radionuclides in soils were within the range of the values obtained in the other parts of the country and within the global average. The ratio of uranium isotopes confirmed the natural origin of uranium. In general, concentrations of heavy metals in the samples of soils, plant leaves, mosses, and lichen are found to be less or in the lower range of values found in other parts of the country, in spite of the differences in plant and moss species or soil characteristics. Possible sources of heavy metal contamination were identified as a power coal plant in the vicinity of the sampling sites and wood and waste burning processes. RECOMMENDATIONS AND PERSPECTIVES: The collected data should provide a base for the health risk assessments on animals and humans in the near future. It should be emphasized that the sampling was carried out 5 years after the military action and that the number of samples was limited; therefore, the conclusions should be accepted only as observed tendencies and a detailed study should be recommended in the future.