Camel trypanosomosis in the Canary Islands: assessment of seroprevalence and infection rates using the card agglutination test (CATT/T. evansi) and parasite detection tests

Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. Thus, the purpose of the present study was to assess seroprevalence and infection rates in the Canary Islands using antibody(-card agglutination test-CATT/T. evansi) and parasite detection tests (micro-Haematocrit Centrifugation technique, Giemsa stained blood smears, microscopic examination of lymph node aspirates and mouse inoculation). PCV was also determined. 745 dromedary camels (483 females and 262 males) were examined. Trypanosomes were detected in seven animals. 36 animals yielded CATT positive results while 709 animals were negative. All parasitologically positive animals were also CATT positive. Results showed a good correlation between CATT positive and low PCV and a higher seroprevalence in older animals. Trypanocidal drugs have not been registered in Spain and, consequently, if vigilance is not exercised the prevalence could be increased in the future.     

Availability and plant uptake of nutrients following the application of paper pulp and lime to tropical acid soils

Availability and plant uptake of nutrients were evaluated in three tropical acid soils (Kandiudult) amended with paper pulp and lime under greenhouse conditions. Amendments were applied to attain target pH values of 5.5, 6.0, and 6.5. A control treatment (no paper pulp or lime added) was also included. Rye grass (Lolium perenne L.) as a test plant was grown for three successive cycles of 40 days each. Extractable nutrients and cumulative nutrient uptake were determined. The application of paper pulp or lime resulted in a significant increase in exchangeable Ca and K and a decrease in exchangeable Mg and extractable Fe, Mn, and Zn. Amendment of soils with paper pulp or lime increased plant uptake of Ca and Mg and decreased that of K, Mn, and Zn. Both amendments behaved similarly, but the effect of lime seemed generally greater than that of paper pulp. Paper pulp in tropical acid soils behaved as a liming agent rather than an organic amendment. Similar to lime, amendment of soils with paper pulp resulted in an increase in availability of Ca and Mg and in a decrease in availability of K, Mn, and Zn for plants. Soil extractions appeared to be appropriate for assessing the availability of Ca, Mn, and Zn. Soil pH and effective cation exchange capacity positively influenced the availability of Ca and negatively the availability of Mn and Zn. Thus, the precision of predicting nutrient availability in paper pulp amended tropical acid soils could be improved by including soil pH or effective cation exchange capacity in relevant regression equations.     

Defining output-based standards to achieve and maintain tuberculosis freedom in farmed deer, with reference to member states of the European Union

Within the European Union (EU), detailed legislation has been developed for cattle, but not deer, to minimise disease risks associated with trade in animals and animal products. This legislation is expressed as input-based standards, providing a detailed outline of the activity required (for example, testing of animals and application of defined control measures), on the expectation that an adequate output (for example, confidence in freedom) will be achieved. Input-based standards are at odds with the increasing shift towards output-based standards, particularly in OIE rules governing international trade. In this paper, we define output-based standards to achieve and maintain freedom from tuberculosis (TB) in farmed deer, with reference to EU member states. After considering the probability of freedom achieved for cattle under existing EU legislation, we defined a ‘free farmed deer holding’ as one with a probability of freedom from infection of at least 99%. We then developed an epidemiological model of TB surveillance systems for deer holdings, incorporating different surveillance strategies, including combinations of diagnostic tests, and a variety of different scenarios relating to the potential for introduction of infection. A range of surveillance strategies were identified to achieve and maintain a free farmed deer holding, and worked examples are presented. The surveillance system sensitivity for varying combinations of screening and confirmatory tests in live animals, animals at slaughter and on-farm deaths is also presented. Using a single test at a single point in time, none of the TB tests routinely used in farmed deer is able to achieve an acceptable probability of TB freedom. If repeat testing were undertaken, an acceptable probability of TB freedom could be achieved, with differing combinations of the surveillance system sensitivity, frequency of testing and risk of introduction. The probability of introduction of infection through the importation of infected deer was influenced by the use of a pre-movement test (assumed 90% test sensitivity and negative test results), the TB prevalence in the source herd and the number of animals imported. A surveillance system sensitivity of at least 81% was achieved with different combinations of annual live animal surveillance and surveillance of animals at slaughter or on-farm deaths. This methodology has broad applicability and could also be extended to other diseases in both deer and other species with relevance to trade in animals and animal products.     

The expression of RoTat 1.2 variable surface glycoprotein (VSG) in Trypanosoma evansi and T. equiperdum

In order to define whether the variable antigenic type RoTat 1.2 is restricted to Trypansoma evansi and could be used as antigen in serological tests to differentiate T. evansi from Trypansoma equiperdum, the appearance of RoTat 1.2-specific antibodies in rabbits, experimentally infected with T. evansi and T. equiperdum, respectively, was analyzed. Ten strains of T. evansi and 11 strains of T. equiperdum originating from Asia, Europe, Africa and Latin America were tested. Rabbit pre-infection sera and sera of days 7, 14, 25, 35 post-infection (p.i.) were analyzed for the presence of antibodies reactive with RoTat 1.2 in immune trypanolysis, ELISA/T. evansi and CATT/T. evansi. Within the duration of the infection (maximum 35 days), all T. evansi as well as 9 out of 11 T. equiperdum infected rabbits became positive in all these tests. The rabbits infected with T. equiperdum OVI (South Africa) and BoTat 1.1 (Morocco) remained negative in the immune trypanolysis test although the latter rabbit became positive in the CATT/T. evansi and ELISA/T. evansi. On the contrary, both rabbits were positive in immune trypanolysis when tested against their respective infecting population. From these data, we conclude that most T. equiperdum strains express isoVATs of RoTat 1.2. This explains, in part, why antibody tests based on T. evansi RoTat 1.2 cannot reliably distinguish between infections caused by T. evansi and those caused by T. equiperdum unless it can be proven that most described T. equiperdum are actually misclassified T. evansi.     

Comparison of serological tests for Trypanosoma evansi natural infections in water buffaloes from north Vietnam

In the present study, a collection of 415 water buffalo serum samples originating from the north of Vietnam was used for evaluation of different diagnostic antibody detection methods available to detect infections with Trypanosoma evansi. The diagnostic sensitivity and specificity of a direct card agglutination test (CATT/T. evansi), an indirect card agglutination test (LATEX/T. evansi) and a newly developed antibody detection ELISA (ELISA/T. evansi) was calculated on the basis of parasitological results, obtained by mouse inoculation, and compared for all assays. The immume trypanolysis assay with the predominant T. evansi RoTat 1.2 variable antigen type was used as reference test for antibody presence. All parasitologically confirmed animals (n=8) were positive in all tests. Diagnostic specificity was highest in CATT/T. evansi (98%) followed by the ELISA/T. evansi (95%) and the LATEX/T. evansi (82%). Concordance of the variant specific immune trypanolysis test with the other tests was calculated and revealed that few (1-8%) false positive results were actually due to a specific reactions, and that LATEX/T. evansi and ELISA/T. evansi detected more immune trypanolysis positives than the CATT/T. evansi. It was concluded that, apart from the immune trypanolysis test, which is not generally applicable, ELISA/T. evansi with a 30% positivity cut-off and LATEX/T. evansi, thanks to their superior capacity of detecting T. evansi specific antibodies, would be suitable as epidemiological tools detecting both active infections and persisting T. evansi specific antibodies. The ELISA/T. evansi with a 50% positivity cut-off and the CATT/T. evansi on the other hand, seem more appropriate to detect true infected water buffaloes.     

Water Extractability of Trace Metals from Soils: Some Pitfalls

Water soluble and/or dissolved metals represent the most ecotoxicologically relevant fraction of metals in the environment. However, water extractions may be prone to errors. This study aims to evaluate the performance of 5 filters as well as Rhizon soil moisture samplers, with respect to metal adsorption and/or release by the filter. In addition, the effect of equilibration time on water extractions of different types of soils was evaluated (silty loam, silty clay loam, loamy sand). Filtrations of synthetic solutions containing 40 μg 1^?1 Zn, 20 μg 1^?1 Cu, Ni and Pb, 10 μg 1^?1 Cr and 2 μg 1^?1 Cd were conducted using the different filters. The synthetic solutions either contained (i) no other competitive cations (Ca, Mg, Na, K, Fe, Mn, Al) (A ₁), (ii) competitive cations at concentrations similar to those observed in soil solutions (A ₂), (iii) competitive cations at 10 times lower concentrations than those in the synthetic soil solution (A ₃). Whiteband filters were observed to retain considerable amounts of trace metals (except Cr), both in the presence and absence of other competitive cations. Millipore filters did not exhibit metal retention. Rhizon soil moisture samplers did not retain trace metals from the synthetic soil solution (A ₂), whereas at lower concentrations of competitive cations (A ₁, A ₃) retention of Cu and Pb was observed. Whiteband filters without a predefined pore diameter allowed colloidal material of unknown particle size to pass into the filtrate, making interpretation of results very difficult and comparison between studies using different filters impossible. Millipore filters with a predefined pore diameter are to be recommended for this purpose. However, particular attention must be paid to potential constitutive Zn release by the sintered glass filter holders, the effect of which can be reduced by rigorous acid washing prior to and following every use. Rhizon samplers were also considered to be useful tools with well-defined and sufficiently small pore diameters to withhold colloidal material. However, in the absence or at reduced concentrations of competitive cations (A ₁, A ₃) retention of trace metals by the Rhizons, particularly Cu and Pb, was observed. Finally, short equilibration times may be insufficient for full assessment of the water extractable pool of trace metals in the soil.     

Use of reactive phosphate rocks as fertilizer on acid upland soils in Indonesia: accumulation of cadmium and zinc in soils and shoots of maize plants

A pot experiment was conducted to study the contribution of reactive phosphate rocks (RPRs) on the accumulation of Cd and Zn in 10 acid upland soils in Indonesia and shoots of Zea mays plants grown on these soils. Two types of RPR were used at a rate of 0.5 g (kg soil)^–1: RPRL containing 4 mg Cd kg^–1 and 224 mg Zn kg^–1, and RPRH containing 69 mg Cd kg^–1 and 745 mg Zn kg^–1. Zea mays was harvested at 6 weeks after planting. The application of RPRH significantly increased the concentrations of Cd in the shoots. The application of this RPR also increased the amount of Cd which could be extracted by 0.5 M NH₄‐acetate + 0.02 M EDTA pH 4.65 from the soils. More than 90% of the added Cd remained in the soil. As Zn is an essential element and the studied acid upland soils are Zn‐deficient, increased plant growth upon RPR application might be partly attributed to Zn present in the phosphate rock. However, more experiments are needed to confirm this hypothesis. The Cd and Zn concentrations and CEC of the soils were important soil factors influencing the concentrations of Cd and Zn in the shoots of maize plants grown on these soils.     

Heavy Metal Fractionation and Extractability in Dredged Sediment Derived Surface Soils

The objective of this study was to characterise pollution with heavy metals in surface soils sampled at various dredged sediment disposal sites in the Flemish region (Belgium). The sites selected varied in the period since sediment disposal ceased and in current vegetation and land use. Total metal contents (Cd, Co, Cu, Fe, Mn, Ni, Pb, and Zn) in the surface soils varied widely. For some disposal sites Cd and Zn concentrations greatly exceeded reference values for clean soil. The distribution of the metals as determined by sequential extraction differed between elements, but was simular for all the soils. This suggested that metals in these sediment derived surface soils were accumulated and transformed in a similar way. Residual fractions were low compared to total contents (2 – 4% for Cd, 25% – 35% for Co, 7 – 18% for Mn, 4 – 22% for Zn, 12 – 41% for Ni, 11 – 42% for Pb, 20 – 45% for Cu, < 10% for Zn). High metal concentrations in the acid-extractable and reducible fractions may indicate pollution from anthropogenic sources. DTPA-extractable metals, which may be considered indicative of plant-available contents, were relatively high compared to the total contents. The relative extractability, expressed as the ratio of DTPA-extractable to total contents, decreased in the order Cd (38%) > Cu (28%) = Zn (26%) > Pb (13%) > Ni (10%) > Co (3%). Most of the sites studied would be of concern if they were used for agricultural activities. No trends in metal availability in the period following disposal were apparent from the data.     

Comparison of serological tests for equine trypanosomosis in naturally infected horses from Kazakhstan

In this study, we compared the complement fixation test (CFT), the horse complement fixation test (HCFT) and a card agglutination test for trypanosomosis (CATT/T. evansi) for the diagnosis of equine trypanosomosis in the Republic of Kazakhstan. Cohen's kappa test was used to evaluate the concordance between the three tests. Kappa scores for CFT versus HCFT and CATT are both 0.6165 (95% Confidence Interval CI 0.414--0.819) indicating a "substantial" agreement between CFT and HCFT or CATT, respectively. Kappa for HCFT versus CATT is 0.395 (CI 0.142--0.648) indicating a "fair" agreement between the two tests. In the absence of a golden standard, seroprevalence and sensitivity and specificity of the three tests were estimated using maximum likelihood estimation. CFT has a sensitivity of 57.2% (CI 31.5--79.5%) and a specificity of 95.8% (CI 89.2--98.5%), HCFT has a sensitivity of 80.6% (CI 44.1--95.6%) and a specificity of 99.5% (CI 90.7--100%), CATT has a sensitivity of 80.2% (CI 44.5--95.2%) and a specificity of 98.5% (CI 79.5--99.9%). The seroprevalence of equine trypanosomosis in Kazakhstan was estimated at 16.4% (CI 9.4--27.0%). The data suggest that for epidemiological studies and the control of equine trypanosomosis serological tests prove useful since they have a high specificity and a satisfactory sensitivity. Field applicable tests, such as CATT/T. evansi may be used to replace laboratory-based tests, such as CFT and HCFT.     

Feasibility of a counter-current extraction procedure for the removal of heavy metals from contaminated soils

Effective remediation and sanitation technologies for soils contaminated with heavy metals are limited. We investigated the feasibility of a counter-current metal extraction procedure for the removal of selected heavy metals (Cd, Cu, Ph, and Zn) from two contaminated soils. The process involved a decarbonation (removal of carbonates), acid solubilisation, washing, and liming step. Results from batch equilibration experiments simulating the counter-current process showed more than 85% of the Cd present to be removed. Removal efficiencies for Cu and Pb were limited to approximately 15%, this mainly due to resorption of these elements during the decarbonation step. As most Zn was found to be present in a more difficult acid-extractable solid phase, its extractability accounted for only 25%. While reaction (pH) conditions of both decarbonation and solubilisation determined removal efficiencies, washing the extracted soil with deionized water only slightly increased the amount of metals removed. Metal distribution among solid phases — exchangeable, carbonate, reducible, organically bound, and residual — was affected by the different treatments. The amount of metals contained in the exchangeable and residual fractions determined their extractability. Except for Cu, the reducible and organically bound fractions were less important. After solubilisation 13 to 70% of the metals were present in an exchangeable solid phase. This implicates that washing the solubilized soil with a salt may increase the extractability of metals, especially for Zn and Pb. Based on our results the process is critically evaluated and possibilities for optimization formulated.